Productive characteristics, chemical composition, in vitro digestibility, and degradation kinetics of two Brachiaria grasses at different regrowth ages.

The study aimed at the effect of different regrowth ages on chemical, productive, and morphological characteristics, in addition to the kinetics of gas production and in vitro digestibility of grasses of the genus Brachiaria. The treatments consisted of two regrowth ages (21 and 35 days) and two grass species (Brachiaria brizantha and B. ruziziensis), in plat with a dimension of 10 × 10 with four replications, totaling 16 plats in a completely randomized design. The regrowth age did not change the leaf:stem ratio of the grasses. Ruziziensis-grass had higher crude protein (CP) content in leaves than Marandu-grass (14.0% versus 10.9% respectively). Marandu-grass leaf had higher NDF content than Ruziziensis-grass (65.0 and 58.3%, respectively) and ADF content (39.6 and 33.2%, respectively). The accumulation rate is high in Marandu-grass regardless of the age of regrowth; however, the in vitro digestibility of dry matter of Ruziziensis-grass is better both in the leaf and in the stem. The regrowth age did not influence the dry matter (DM) and neutral detergent fiber (NDF) in vitro digestibility (ivD) of the leaves. The ivDNDF of leaves of both types of grass were similar. There was an increase in the ivDDM of stem when the regrowth age was 21 days. The DM degradation rate was higher in Marandu-grass at both regrowth ages, and the total gas production was higher at 21 days. The parameters evaluated in the dual-pool logistic model showed interaction for age and cultivar, except for fractions λ (lag time) and total gas production. Marandu-grass presented a greater volume of gas for the rapidly degradable fraction and a lower degradation rate. There was a significant interaction for the parameters evaluated in the models between ages and grasses. The exponential model showed interaction in all fractions. Grasses had reduced CP content and increased fibrous fraction as a function of age, with no reduction in leaf DM and NDF digestibility. The Ruziziensis-grass has higher digestibility at the regrowth ages evaluated, while Marandu-grass has higher yields. We recommend adjusting the in vitro degradation kinetics by the dual-pool logistic model.

A study of the structure-activity relationship of oligomeric ellagitannins on ruminal fermentation in vitro.

The aim of this study was to investigate how the degree of oligomerization of ellagitannins (ET) influences their ability to alter ruminal fermentation. Dimeric to heptameric ET were isolated from rosebay willowherb (Epilobium angustifolium) flowers and purified. Ellagitannins were tested in vitro on a mixture of grass silage and buffered rumen fluid. Total gas production was measured in real time using an automated pressure evaluation system. Methane production was monitored at regular interval by gas chromatography for 72h. The effect of ET was evaluated on 2 sources of rumen fluid using a randomized block design. Ammonia nitrogen, volatile fatty acid concentration, and pH were measured at the end of the experiment. Results show that oligomeric ET decreased gas production and total volatile fatty acid concentration proportionally to their degree of oligomerization. Methane production was also decreased by all the tested compounds and dimer was less effective than the larger ET, which showed similar levels of activity. Additionally, willowherb's oligomeric ET decreased ammonia-nitrogen and branched-chain volatile fatty acid concentrations, thus indicating reduced protein degradation by ruminal bacteria. This effect showed a quadratic relationship with the degree of oligomerization and was maximal with the tetramer. In conclusion, this study shows that the degree of oligomerization of ET has more than a simple linear effect on fermentation parameters in vitro. Large oligomers, in fact, have more detrimental effects on volatile fatty acid and gas production than small ones, while being similarly effective at inhibiting methane production.

Standardisation of the C:N ratio in ileal digesta changes relationships among fermentation end-products during in vitro hindgut fermentation in pigs.

Undigested proteins that become available for the microbiota in the hindgut can be used as building blocks for bacterial cells, or can enter various catabolic pathways. Degradation via protein fermentation pathways is least preferred, as several fermentation end-products released can be toxic for the host. Directing microbial protein metabolism towards protein synthesis or degradative pathways that result in less toxic end-products, for example through nutritional interventions, is an interesting strategy for improving health. We studied variation in protein fermentation patterns, resulting from variation in substrate composition. Ileal digesta, obtained from cannulated pigs fed different protein sources, were subjected to fermentation in vitro under different conditions; (1) ileal digesta were fermented as-is, (2) ileal digesta were fermented after standardisation to a constant high C:N ratio, by addition of high fermentable carbohydrates and (3) ileal digesta samples were incubated under limiting N concentrations. Gas production was monitored as an indirect measure of microbial activity, and fermentation end-products at different points in time were analysed by gas chromatography and high resolution mass spectrometry. Using principal component analysis, we identified patterns in protein fermentation end-products and related them to the composition of ileal digesta. Protein-associated fermentation end-product concentrations of e.g. isovaleric-, isobutyric-, phenylacetic acid and p-cresol were negatively affected by the available amount of high fermentable carbohydrates combined with a high C:N ratio. The aforementioned fermentation end-products positively correlated with NH3 concentrations and negatively with short-chain fatty acid (SCFA) concentrations. Standardisation to a constant high C:N ratio changed their relationship; isovaleric-, isobutyric-, phenylacetic acid and p-cresol lost their correlation with NH3 concentrations, became positively correlated with SCFA concentrations, and now showed a positive correlation with available amounts of high fermentable carbohydrates. Our observations demonstrate an important role of the C:N ratio in the relationship between fermentation end-products. At constant C:N, protein fermentation end-products correlate with end-products of carbohydrate fermentation and NH3, often considered as a proxy for protein fermentation, loses its predictive power.

Chemical Characterization and In Vitro Gas Production Kinetics of Alternative Feed Resources for Small Ruminants in the Maltese Islands.

The ever-increasing human population, the problem associated with climate change and recent crises-COVID-19 disease and trade conflicts-all impacted on the availability and cost of animal feed raw materials. This is clearly visible in realities which heavily rely on importation such as islands and small states, where producers involved in the agricultural sector were strongly affected by the sharp increase in prices. To deal with these global issues, alternative resources are perceived to replace conventional ingredients. This work aimed at assessing the nutritive value of different resources (sheep feed, mature carob, Maltese bread, wild asparagus, prickly lettuce, and loquat) for small ruminants present in the Maltese Islands, analyzing their chemical composition, gas production kinetics and antioxidant properties. In general, the variation in chemical composition resulted in different rumen fermentation kinetics (p < 0.007). The ratio between GP-24 h and GP-48 h was higher in Maltese bread than other substrates; loquat, prickly lettuce and wild asparagus showed lower fermentation kinetics in accordance with their high NDF and ADF contents. The antioxidant activity may be partially related to the polyphenolic content that was higher in wild asparagus, prickly lettuce and loquat. All feed characteristic confirmed their potential to be included as ingredients in ruminant diets and as a source of fiber.

An in vitro model for caecal proteolytic fermentation potential of ingredients in broilers.

Fermentation of protein in the caeca of chickens may lead to the production of potentially detrimental metabolites, which can reduce gut health. A poor precaecal digestion is expected to increase protein fermentation (PF), as more proteins are likely to enter the caeca. It is unknown if the undigested protein that enters the caeca differs in fermentability depending on their ingredient source. In order to predict which feed ingredients increase the risk of PF, an in vitro procedure was developed, which simulates the gastric and enteric digestion, subsequent caecal fermentation. After digestion, amino acids and peptides smaller than 3.5 kD in the soluble fraction were removed by means of dialysis. These amino acids and peptides are assumed to be hydrolysed and absorbed in the small intestine of poultry and therefore not used in the fermentation assay. The remaining soluble and fine digesta fractions were inoculated with caecal microbes. In chicken, the soluble and fine fractions enter the caeca, to be fermented, while insoluble and coarse fractions bypass them. The inoculum was made N-free to ensure bacteria would require the N from the digesta fractions for their growth and activity. The gas production (GP) from the inoculum, therefore, reflected the ability of bacteria to use N from substrates and was an indirect measure for PF. The Maximum GP rate of ingredients averaged 21.3 ± 0.9 ml/h (mean ± SEM) and was in some cases more rapid than the positive control (urea, maximum GP rate = 16.5 ml/h). Only small differences in GP kinetics were found between protein ingredients. Branched-chain fatty acids and ammonia concentrations in the fermentation fluid after 24 hours showed no differences between ingredients. Results indicate that solubilised undigested proteins larger than 3.5 kD are rapidly fermented independent of its source when an equal amount of N is present.

An Illustrative Analysis of Atypical Gas Production Profiles Obtained from In Vitro Digestibility Studies Using Fecal Inoculum.

Gas production profiles typically show a monotonically increasing monophasic pattern. However, atypical gas production profiles exist whereby at least two consecutive phases of gas production or additional extraneous features that distort the typical profile are present. Such profiles are more likely to occur with the use of a fecal inoculum and are much less well described. The presence of multiple phases or non-descript extraneous features makes it difficult to apply directly recommended modeling approaches such as standard response functions or classical growth functions. To overcome such difficulties, extensions of the Mitscherlich equation and a numerical modeling option also based on the Mitscherlich are explored. The numerical modeling option uses an estimate of relative rate obtained from the smoothed data profile and an estimate of maximum gas produced together with any lag time information drawn from the raw data to construct a simple Mitscherlich equation. In summary, this article illustrates the analysis of atypical gas production profiles obtained using a fecal inoculum and explores the methodology of numerical modeling to reconstruct equivalent typical growth-like trends.

Different processing effects on nutritive value of barely grains by gas production method.

The purpose of this study was to investigate the effects of warm and cold physical as well as chemical processing methods on the nutritive value of barley grains by gas production technique. The processing methods included milling, steam flaking, extruding and soaking up the grain in water containing 1.00% citric acid, 1.00% propionic acid and 0.01 M sulfuric acid. Two-hundred mg of milling samples were incubated in special 100 mL glasses and the amount of gas produced at different hr was measured. The data were analyzed in a completely randomized design. The results showed that different treatments did not have a significant effect on chemical composition of barley seeds. Other methods of processing compared to the grinding method, significantly reduced the total amount of produced gas. In the 72 hr incubation period, the lowest amount of gas production was in the extruded (245.6 mL per g dry matter) treatment. However, there was no significant difference between the two methods of extruding and flaking. The highest percentage of digestible organic matter was associated with propionic acid (64.90%), while the steam cracking method (58.74%) was the least. Among the processing methods, the highest amounts of methane production, total protozoa population and volatile fatty acid concentration were related to the grinding method and the least amount of extrusion treatment was observed during 24 hr of incubation. Different experimental treatments had a significant effect on ammonia nitrogen condensation and the highest level was observed in milling. According to our results, processing methods such as extrusion and flaking may improve the grain nutritive value.

Models Based on the Mitscherlich Equation for Describing Typical and Atypical Gas Production Profiles Obtained from In Vitro Digestibility Studies Using Equine Faecal Inoculum.

Two models are proposed to describe atypical biphasic gas production profiles obtained from in vitro digestibility studies. The models are extensions of the standard Mitscherlich equation, comprising either two Mitscherlich terms or one Mitscherlich and one linear term. Two models that describe typical monophasic gas production curves, the standard Mitscherlich and the France model [a generalised Mitscherlich (root-t) equation], were assessed for comparison. Models were fitted to 25 gas production profiles resulting from incubating feedstuffs with faecal inocula from equines. Seventeen profiles displayed atypical biphasic patterns while the other eight displayed typical monophasic patterns. Models were evaluated using statistical measures of goodness-of-fit and by analysis of residuals. Good agreement was found between observed atypical profiles values and fitted values obtained with the two biphasic models, and both can revert to a simple Mitscherlich allowing them to describe typical monophasic profiles. The models contain kinetic fermentation parameters that can be used in conjunction with substrate degradability information and digesta passage rate to calculate extent of substrate degradation in the rumen or hindgut. Thus, models link the in vitro gas production technique to nutrient supply in the animal by providing information relating to digestion and nutritive value of feedstuffs.

Chemical Composition, In Vitro Digestibility and Rumen Fermentation Kinetics of Agro-Industrial By-Products.

The nutritive value of 26 agro-industrial by-products was assessed from their chemical composition, in vitro digestibility and rumen fermentation kinetics. By-products from sugar beet, grape, olive tree, almond, broccoli, lettuce, asparagus, green bean, artichoke, peas, broad beans, tomato, pepper, apple pomace and citrus were evaluated. Chemical composition, in vitro digestibility and fermentation kinetics varied largely across the by-products. Data were subjected to multivariate and principal component analyses (PCA). According to a multivariate cluster analysis chart, samples formed four distinctive groups (A-D). Less degradable by-products were olive tree leaves, pepper skins and grape seeds (group A); whereas the more degradable ones were sugar beet, orange, lemon and clementine pulps (group D). In the PCA plot, component 1 segregated samples of groups A and B from those of groups C and D. Considering the large variability among by-products, most of them can be regarded as potential ingredients in ruminant rations. Depending on the characteristic nutritive value of each by-product, these feedstuffs can provide alternative sources of energy (e.g., citrus pulps), protein (e.g., asparagus rinds), soluble fibre (e.g., sugar beet pulp) or less digestible roughage (e.g., grape seeds or pepper skin).

Rumen liquor from slaughtered cattle as inoculum for feed evaluation.

Use of nonlinear mathematical models has been majorly based on in vitro gas production (GP) data generated when substrates are incubated with rumen liquor from fistulated steers. However, existing evidence suggests that rumen liquor from slaughtered cattle of unknown dietary history also generates quantifiable in vitro GP data. Fitting and description of GP data obtained from 4 diets incubated with rumen liquor from slaughtered cattle was evaluated using single-pool exponential model with discrete lag time (EXPL), logistic (LOG), Groot's (GRTS) and Gompertz (GOMP) models. Diets were formulated by varying proportions of Rhodes grass (Chloris gayana) hay and a concentrate mixed on dry matter basis to be: 1,000 g/kg Rhodes grass hay (RGH) and 0 of the concentrate (D1), 900 g/kg RGH and 100 g/kg concentrate (D2), 800 g/kg RGH and 200 g/kg concentrate (D3), 700 g/kg RGH and 300 g/kg concentrate (D4). Dietary kinetics for the models were determined by measuring GP at 2, 4, 8, 10, 18, 24, 36, 48, 72, 96 and 120 h. Model comparison was based on derived GP kinetics, graphical analysis of observed versus predicted GP profiles plus residual distribution and goodness-of-fit from analysis of root mean square error (RMSE), adjusted coefficient of determination (Adj-R2) and Akaike's information criterion (AIC). Asymptotic GP, half-life and fractional rate of GP differed (P < 0.001) among the 4 models. The RMSE, Adj-R2 and AIC ranged from 1.555 to 4.429, 0.906 to 0.984 and 2.452 to 15.874, respectively, for all diets compared across the 4 models. Based on the goodness-of-fit statistical criterion, GP profiles of D1 were more appropriately fitted and described by GRTS and GOMP than the EXPL and LOG models. The GRTS model had the lowest AIC value for D2 (2.452). Although GRTS model had the most homogenous residual dispersion for the 4 diets, all the 4 models exhibited a sigmoidal behavior. Therefore, rumen liquor from slaughtered cattle of unknown dietary history can be used to derive nutritionally important feed parameters, but choice of the most appropriate model should be made based on fitting criteria and dietary substrates incubated.

In vitro gas production in rumen fluid of buffalo as affected by urea-calcium mixture in high-quality feed block.

This study aimed to determine the effect of urea-calcium sulphate mixture (U-cas) levels in high-quality feed block (HQFB) on ruminal digestibility, fermentation and gas kinetics in rumen fluid of swamp buffalo by using in vitro techniques. The treatments were seven levels of U-cas incorporated in HQFB at 0, 3, 6, 9, 12, 15 and 18% and the experimental design was a completely randomized design. Gas production rate constants for the insoluble fraction, potential extent of gas and cumulative gas were linearly increased with increasing levels of U-cas in HQFB. The in vitro dry matter digestibility, in vitro organic matter digestibility, true digestibility and microbial mass were altered by treatments and were greatest at 18% U-cas supplementation. Concentrations of propionate were linearly increased with increasing levels of U-cas and was highest with U-cas supplementation at 18%. The NH3 -N concentration was highest when urea was added in the HQFB while NH3 -N concentration tended to be reduced with increasing level of U-cas. The findings suggest supplementation of 18% U-cas in HQFB improves kinetics of gas production, rumen fermentation, digestibility and microbial mass as well as controlling the rate of N degradation in the rumen of swamp buffalo.

Efficacy of different methanolic plant extracts on anti-methanogenesis, rumen fermentation and gas production kinetics in vitro.

The present study was carried out to evaluate the effect of methanolic extracts of three plants, mehandi (Lawsonia inermis), jaiphal (Myristica fragrans) and green chili (Capsicum annuum) on methanogenesis, rumen fermentation and fermentation kinetic parameters by in vitro gas production techniques. Single dose of each plant extract (1 ml / 30 ml buffered rumen fluid) and two sorghum fodder containing diets (high and low fiber diets) were used for evaluating the effect on methanogenesis and rumen fermentation pattern, while sequential incubations (0, 1, 2, 3, 6 9, 12, 24, 36, 48, 60, 72 and 96 h) were carried out for gas production kinetics. Results showed that methane production was reduced, ammonia nitrogen was increased significantly, while no significant effect was found on pH and protozoal population following addition of different plant extracts in both diets except mehandi. Green chili significantly reduced digestibility of dry matter, total fatty acid and acetate concentration at incubation with sorghum based high and low fiber diets. Among all treatments, green chili increased potential gas production, while jaiphal decreased the gas production rate constant significantly. The present results demonstrate that methanolic extracts of different plants are promising rumen modifying agents. They have the potential to modulate the methane production, potential gas production, gas production rate constant, dry matter digestibility and microbial biomass synthesis.

Cinética de fermentação ruminal de silagens de milheto / Ruminal fermentation kinetics of pearl millet silages

Avaliaram-se a degradabilidade da matéria seca e a cinética de fermentação ruminal das silagens de três genótipos de milheto (BRS-1501, NPM-1 e CMS-3), por meio da técnica in vitro semi-automática de produção de gases. O delineamento experimental utilizado foi o de blocos ao acaso em arranjo de parcelas subdivididas. Para a comparação dos genótipos, foi utilizado o teste SNK (P<0,05), e os parâmetros da cinética de fermentação foram ajustados ao modelo unicompartimental de France. A partir de 24 horas, a silagem do genótipo BRS-1501 foi a que apresentou maior produção acumulativa de gases. Os valores de degradabilidade da matéria seca apresentados às 96 horas de fermentação foram de 53,9 por cento para a silagem do BRS-1501, 51,7 por cento para a silagem do CMS-3 e 49,1 por cento para a silagem do NPM-1, sendo este o material com menor degradabilidade (P<0,05). Os potenciais máximos de produção de gases variaram de 145,18 para o genótipo CMS-3 a 155,02ml/g de MS para o BRS-1501. A silagem do genótipo BRS-1501 destacou-se entre as demais, em função do seu maior potencial de produção de gases e degradabilidades efetivas.

The dry matter digestibility and the fermentation kinetics of three pearl millet genotypes silages (BRS-1501, NPM-1, and CMS-3) were estimated by the semi-automated in vitro gas production technique, using a completely randomized blocks design in a split plot arrangement. The genotypes means were compared by SNK test (P<0.05) and fermentation kinetics parameters estimated by regression of gas production on time using the France model. The greater cumulative gas production from 24 hours on was obtained by BRS-1501 genotype silage. The dry matter degradability (DMD) values at 96 hours of incubation were 53.9 percent for BRS-1501, 51.7 percent for NPM-1, and 49.1 percent for CMS-3 silage. The lowest DMD (P<0.05) among the studied genotypes silages was observed for CMS-3 silage. The maximum potential of gas production ranged from 145.18ml/g of dry matter for CMS-3 genotype silage to 155.02ml/g of dry matter for BRS-1501 silage. The BRS-1501 genotype silage showed the highest potential of gas production and effective degradabilities.