Buffered Rumen Liquor Research Articles

In vitro gas fermentation kinetics of compounded diets containing varying levels of Tetrapleura tetraptera fruit meal

The study investigated in vitro gas fermentation kinetics of compounded diets containing different quantities of Tetrapleura tetraptera (TTFM). Three replicates of each dietary sample (200 mg) were incubated in buffered rumen liquor for 48 hours and the parameters such as in vitro dry matter digestibility (IVDMD), organic matter digestibility (OMD), short-chain fatty acids (SCFA) and methane gas (CH4) were determined using in vitro gas production technique. One-way Analysis of Variance (ANOVA) was used to analyze the gathered data. The IVDMD exhibited substantial variation (p<0.05) among treatments. The diet with 1.0 % Tetrapleura tetraptera had the highest in vitro dry matter digestibility (IVDMD), which may be useful for effective feed conversion efficiency. The acetate and propionate with total volatile fatty acids raised the inclusion levels of TTFM in the rams' diet by 1.5 to 2.0 %. Tetrapleura tetraptera fruit meal can be added to ruminant diets at a rate of up to 2.0 % to increase daily weight gain, nitrogen utilization, and nutrient digestibility, resulting in better storage of energy

Effect of replacing corn grains with date palm kernels on ruminal fermentation, feed degradability, and methane production under different initial in vitro pH conditions

Date kernels (DK) are cheap by-products rich in energy and phenolic compounds. It can be used as an alternative to the conventional sources of energy in ruminant diets while reducing methane (CH4) production. Using a semi-automated gas production (GP) system, the initial pH of buffered rumen liquor was adjusted to 5.5 and 6.8. Five experimental diets were evaluated, control (0% DK), and DK25, DK50, DK75, and DK100 presented 25, 50, 75, and 100% replacement of maize by DK, respectively. Of the 16 phenolic compounds detected in DK, protocatechuic, p-hydroxybenzoic and catechin were the most abundant. At pH 6.8, the control diet recorded higher (p < 0.05) GP values throughout the first 12 h incubation than all other DK diets, while at 5.5 pH, DK50 displayed the highest (p < 0.05) GP at 3 and 6 h compared to all other diets. At either pH conditions, all DK diets reduced (p < 0.05) CH4 compared to the control without affecting protozoal counts. At 5.5 pH, DK diets showed enhanced (p < 0.05) nutrients degradability compared to control. DK modified (P < 0.05) the fermentation patterns toward more propionate than the control under either pH conditions. Substitution of maize by 50% DK was highly recommended in ruminant diets.

Preliminary in vitro screening of some spices and medicinal plants from Edo and Rivers States, Nigeria for reducing enteric methane production in ruminants

Methane gas produced by ruminants during feed fermentation contributed to global warming as well as poor efficiency of food utilization. Mitigating ruminal methane production through anti-microbial feed additive has serious health implications. A preliminary study of the potential of medicinal plants and spices from Edo and Rivers State, as possible replacement to antibiotic feed additives was carried out. Medicinal plants and spices were purposefully sampled, dried at 40oC, milled and 10 mg was added with 190 mg of substrate in a preweighed nylon incubation bags. Bags and their contents were incubated using 30 mL of buffered rumen liquor in 100 mL syringes following the standard procedure of in vitro fermentation. Variables monitored were 24h total gas volume, short chain fatty acids (SCFA) methane (CH4 ) gas, dry matter degradability (DMD) as well as some chemical, cell wall and anti-nutritive components. Results revealed that adding the medicinal plants and spices as feed additives did not depress gas production in most cases (19.5 – 29.50 mL, 17.75 – 30.00 mL and 12.00 – 29.00 mL for Edo medicinal plants, Edo spices and Rivers medicinal plants respectively) and also did not depress DMD (&gt; 50%). Azadirachta indica stem reduced CH4 production mostly by 53.56% while Dennttia tripetala did by 47.69% among the Edo medicinal plants and spices, respectively. In Rivers State, Amaranthus spinosus gave the highest value of 60.33% CH4 reduction. In the same bracket of similar and good CH4 reduction potentials were Alstonia boonei leaves (49.06%), Newbouldia laevis root (41.50%), Euphorbia heterophyla (37.97%), Allium crispum (47.31%), Allium cepa (46.80%), Aframomum mueguata (46.54%), Capsicum annum (47.32%), Ageratum conyzoides (56.52%), Lagenaria breviflora (48.03%) and Centrosema molle (45.08%). All the medicinal plant and spices with potential for CH4 reduction had corresponding high SCFA values (0.46 – 0.65 Mmol) with tannin (%) and saponin (%) contents of 0.011 – 17.50 and 0.01 – 2.40 respectively. Conclusively, the medicinal plants and spices demonstrated good potentials for reducing rumen methane production in vitro and may be subjected to further in vivo studies.

In vitro gas fermentation kinetics of compounded ration containing graded levels of biodegraded corncob

Cob regarded as a waste product, constituting source of environmental pollution could be utilized as an energy source in feed formulation if properly processed and harnessed through fungal degradation. Hence, this study was designed to evaluate the nutritive value of biodegraded corncob meal based diet using in vitro gas production technique. Aspergillus niger was isolated and sub-cultured to obtain a pure culture. Degraded corn cob meals(DCCM) of four dietary treatments were prepared to include: T (0% DCCM which served as 1 the control), T (15% DCCM), T (30% DCCM) and T (45% DCCM). Each diet sample 2 3 4 (200mg) was incubated in buffered rumen liquor for 48 hours and gas volume was estimated using established in vitro gas production models. Amount of gas volume produced was determined every 3 hours for 48 hours of incubation in buffered rumen fluid. After 48 hours ofincubation, methane gas produced was estimated and determined in triplicates. The proximate composition of Aspergillus niger biodegraded corn cob indicated that Aspergillus niger improved the nutritive value of the corn cob. Untreated corn cob meal was lower in nutritive values when compared with treated corncob meal with crude protein, crude fibre and ash values ranging from 6.88 to 9.78%, 32.68 to 26.37% and 2.87 to 2.88% respectively.The proximate composition of the dietary treatments showed that the crude protein varied from (11.67-12.67%), crude fibre (10.94-21.56%), ether extract (2.12-4.88%), ash (6.48- 9.44%) and nitrogen free extract (58.17-62.99%). Results obtained for volume of gas (35.960-72.770mL/200mgDM) produced in time “t” denoted by (b) were significantly different (P&lt;0.05) across the dietary treatments. However, rate of gas production (0.0297- 0.0425mL/hr) and time between incubation and gas production (2.083-2.683hr) were not significantly different (P&gt;0.05) across the dietary treatments. Cumulative gas volumeproduction at 24 hours were significantly (P&lt;0.05) influenced by different inclusion levels of DCCM with values obtained ranging from T (15.33 ml/gDM) to T (35.33 ml/gDM). There 4 3 were significant differences (P&lt;0.05) across the dietary treatments. The post estimated parameters ranging from metabolisable energy (4.96-7.74MJ/Kg DM), organic matter digestibility (37.98-58.03%), short chain fatty acids (0.31-0.78μmol), Methane estimate (3-8 ml/200mgDM) and Carbon dioxide estimate (10-30 ml/200mgDM). It can be concluded that 30% DCCM based diet had the potential of meeting the nutritional needs as smallruminant livestock feeds, if properly biodegraded and incorporated into feeds.

Effects of polyphenol rich bamboo leaf on rumen fermentation characteristics and methane gas production in an in vitro condition

The aim of this study was to test the effect of bamboo leaf (BL) on rumen methane gas production and rumen fermentation characteristics, in vitro. Different amounts of BL; CON (0 %), Low BL (LBL, 10 %), Medium BL (MBL, 15%) and High BL (HBL, 25%) of replacement with alfalfa hay (AH) in substrate (50 % concentrate + 50 % AH) were mixed with 30 millilitre (mL) of buffered rumen liquor for 48 h of incubation. Total gas production (mL/250 mg DM) was not affected (P&gt;0.05) among BL treatment groups at different times of incubation. Production of methane gas (mL/250mg DM) decreased at a declining rate (P less than 0.05) with higher BL levels. Methane gas inhibitory effects of BL treatment groups as compared with CON were; 29%, 35% and 62% for LBL, MBL and HBL, respectively. The ratio of acetic/propionic was lowest (P less than 0.05) for HBL (1.67) as compared to CON (2.09).

Inclusion of Different Levels of Eucalyptus Oil: An In Vitro Trial to Study the Effect on Sheep Rumen Methanogenesis and Fermentation Processes

An in vitro trial was conducted in fermentation vessels under anaerobic conditions using composite feed-1 and composite feed-2 with inclusion of different levels of Eucalyptus oil (EO) along with buffered rumen liquor of sheep to observe the effects of oil on rumen methanogenesis and fermentation. The CF-1 consisted of 40% of concentrate and CF-2 had 25 % of concentrate. The various concentrations of EO used in present study were 0.5,1 and 2 µl/ml. During the present investigation it was observed that on increasing concentration of EO in CF-1 and CF-2 resulted in linear decrease in total gas production up to 74.26% in CF-1; and 31.01% by inclusion of EO in CF-2, respectively. The findings suggest that decrease in total gas production and methane concentration was highly pronounced in CF-1 compared to CF-2. The rumen digestibility in CF-1 and CF-2 also decreased significantly (p &lt; 0.01) from control. A significant decrease in NH3 –N compared to control was also documented. As EO in both the feeds was found to inhibit deamination activity, the ammonia production decreased with increase in the efficiency of protein utilization. The inclusion of the oil decreased the Total Volatile Fatty Acids (TVFA) concentration significantly (p &lt; 0.01). The A:P ratio increased by inclusion in CF-2 but not in CF-1. The pH was effected on inclusion of EO in CF-2 whereas inclusion of EO in CF-1 exhibited non significant effect. Therefore, the difference in Volatile Fatty acids (VFA) pattern due to essential oil might be because of different diets and pH in different experiments. The inclusion of EO decreased the protozoa number significantly (p &lt; 0.01). EO was found to decrease fermentation in a manner that even at 0.5 µl/ml concentration there was a significant decrease in GP, ME, SCFA, TDDM and MBP. It could therefore be envisaged that increasing the proportion of concentrate in diets and using EO as a feed additive increases the rumen efficiency by reducing the methane concentration and gas production.

Effect of replacing oat fodder with fresh and chopped oak leaves on in vitro rumen fermentation, digestibility and metabolizable energy.

Aim:A study was conducted to evaluate the effect of replacing oat fodder (OF) with fresh oak leaves (FOL) or chopped oak leaves (COL) on rumen fermentation and digestibility through in vitro gas production technique (IVGPT).Materials and Methods:Nine different diets were prepared by mixing OF with oak leaves (either FOL or COL) in different ratios (100:0, 75:25, 50:50, 25:75, and 0:100). The rations were evaluated through Hohenheim IVGPT with 200 mg substrate and 30 ml of buffered rumen liquor. All the syringes were incubated at 39°C for 24 h in buffered rumen liquor of cattle. After 24 h, the total gas production was recorded, and the contents were analyzed for in vitro methane production, protozoa no. and ammonia-N.Results:Chopping (p<0.01) reduced the tannin fractions as well as non-tannin phenol. Increase in levels of oak decreased total gas production, methane, organic matter (OM) digestibility, and metabolizable energy (ME) values. The polyphenol content of the substrate did not show any significant difference on the protozoal count.Conclusion:In vitro studies revealed that the addition of oak leaves reduced the methane production and ammonia nitrogen levels; however, it also decreased the OM digestibility and ME values linearly as the level of the oak leaves increased in the diet. Chopping was effective only at lower inclusion levels. Further studies, especially in vivo studies, are needed to explore the safe inclusion levels of oak leaves in the diet of ruminants.

An in vitro trial to study the effect of inclusion of different levels of coconut oil on sheep rumen methanogenesis and fermentation processes

An in vitro trial was conducted in fermentation vessels under anaerobic conditions using composite feed-1 and composite feed-2 with inclusion of different levels of Coconut Oil (CO) along with buffered rumen liquor of sheep to observe the effects of oil on rumen methanogenesis and fermentation. The Composite Feed-1(CF-1) consisted of 40% of concentrate and Composite Feed-2 (CF-2) had 25 % of concentrate. The different levels of CO used were 0.5,1 and 2 µl/ml. Among the gas production and methane concentration parameters the inclusion of CO showed a significant decrease of 38.67%. A non significant effect on gas production was noted with the inclusion of CO in CF-2. Further, a decrease in total gas production and methane concentration was observed which was highly pronounced in CF-1 as compared to CF-2. The rumen digestibility by inclusion of CO in CF-1 and CF-2 decreased significantly (P &lt; 0.01) from control. The inclusion of CO in CF-2 did not affect Ammonia Nitrogen (NH3 –N) concentration, while in CF-1 the CO inclusion increased the NH3 –N concentration. The inclusion of CO decreased the Total volatile fatty acid (TVFA) concentration significantly (P &lt; 0.01). In both the diets namely CF-1 and CF-2 the A:P ratio was unaffected by CO inclusion. The inclusion of CO in CF-1 had a non significant effect in pH. However, the inclusion of CO in CF-1 and CF-2 decreased the protozoa number significantly (P &lt; 0.01). Inclusion of oil tended to decrease Gas production (GP), Metabolizable energy (ME), Short Chain Fatty Acid (SCFA), True Degradability of Dry Matter (TDDM) and Microbial Biomass Production (MBP). The effect was more pronounced when CO level was 1.0µl/ml or higher. From the above gas production and fermentation parameters noticed in two diets CF-1 and CF-2 with inclusion of different levels of CO, it can be envisaged that increasing the proportion of concentrate in diets and using CO as a feed additive increases the rumen efficiency by reducing the methane concentration and gas production.

Changes in fatty acid composition of various full fat crushed oilseeds and their free oils when incubated with rumen liquor in vitro

The fatty acid pattern of dietary lipids can be modified during rumen biohydrogenation (BH). The objective of the present study was to assess changes in the FA pattern of different oilseed products supplied either as crushed full fat oilseed or as free oil after in vitro incubation with buffered rumen liquor. The FA patterns were determined at the beginning and compared with those measured after 24 h of incubation. The contents of fatty acids (FA) < C18 increased (p < 0.05) in nearly all treatments, eventually due to microbial de novo synthesis and fermentation of carbohydrates and proteins during incubation. In contrast, the contents of the dominating C18 FA, (oleic acid – C18:1c9, linoleic acid – C18:2c9,12, linolenic acid – C18:3c9,12,15) were reduced due to BH, resulting in the accumulation of characteristic BH intermediates, such as conjugated linoleic acid (CLA) isomer C18:2c9t11 (rumenic acid). However, both for crushed full fat oilseeds and their free oils the process of BH was not completed at the end of incubation. The disappearance was highest for C18:3c9,12,15, followed by C18:2c9,12 and C18:1c9. The rate of BH of unsaturated FA was higher in the crushed form compared to the oil form. Higher amounts of BH intermediates accumulated in the crushed form. Obviously, the physical form affects the degree of BH in vitro. The current results suggest that feeding crushed full fat seeds instead of their free oils to dairy cows might stimulate the formation of beneficial BH intermediates such as CLA in the rumen.

RUMINAL FERMENTATION KINETICS OF MORINGA AND PELTIPHYLLUM SUPPLEMENTS DURING EARLY INCUBATION PERIOD IN THE IN VITRO READING PRESSURE TECHNIQUE

This experiment was aimed to observe rumen fermentation kinetics of alternative supplements, i.e.Moringa oleifera and Peltiphyllum peltatum leaves added to maize silage diet as compared to aconventional supplement (barley-soya). A total of six treatments were investigated in the present study,which consisted of: maize silage (A), M. oleifera (B), P. peltatum (C), maize silage + concentrate (56:44,w/w; D), maize silage + M. oleifera (56:44, w/w; E), and maize silage + concentrate + P. peltatum(47:37:16, w/w/w; F). The feeds were incubated in vitro in three replicates (completely randomized)using the Reading Pressure Technique system. Approximately 800 mg of the feeds were mixed by 75 mlof buffered rumen liquor. The incubation was carried out up to 12 h in bottles and gas pressure wasrecorded and released in regular intervals. Repeated sampling was conducted for analysis offermentation products at 1, 6 and 12 h of incubation period. The results showed that the nutritionalquality of Moringa and its fermentation pattern was comparable to that of barley-soya concentrate. PlainMoringa incubation resulted the highest production of iso-SCFA and soluble protein concentrationamong all treatments after 12 h incubation (P<0.05). Incubation of sole Peltiphyllum peltatum highlyhampered the fermentation rate. Production of iso-SCFA both for plain Peltiphyllum and its mixturewere comparatively low. Supplementation of Peltiphyllum increased significantly soluble proteinconcentration during 12 h incubation (P<0.05). Peltiphyllum also had a very low C2+C4/C3 ratiocompared to other treatments. It could be concluded that Moringa oleifera is a potential alternativesupplement to replace either partially or completely concentrate as a conventional supplement, andPeltiphyllum peltatum supplementation could reduce excessive protein degradation and fermentation ofthe concentrate in the rumen.

Germination temperature and time affect in vitro fermentability of sorghum grain

Two experiments were conducted to determine the effect of temperature of steeping or soaking (18 or 25 °C) and germination time (0, 1, 3 or 5 days) (Experiment 1), and the effect of germination temperature (18, 25 or 32 °C) and germination time (1, 3 or 5 days) (Experiment 2) on the in vitro fermentability of sorghum. Dry rolling of grain was used as the control treatment in Experiment 1. Fermentability was evaluated by measuring the amount of starch that was fermented and the total volatile fatty acids (VFA) produced when grain was incubated in buffered rumen liquor for 5 h. Compared to dry rolling, increasing germination time to 5 days significantly increased the amount of starch fermented by 13% (P<0.01) and the total VFA production by 34% (P<0.001) (Experiment 1). Germination time had significant interaction with germination temperature (Experiment 2) but not with steeping temperature (Experiment 1). Germination time had a quadratic effect on the fermentability of germinated sorghum and the slope was different for each temperature of germination. Total VFA production was higher (P<0.05) for sorghum germinated at 25 or 32 °C (35–46 mmol L −1) than for sorghum germinated at 18 °C (29–40 mmol L −1). Changing the temperature during steeping did not affect (P>0.05) the amount of starch fermented (P=0.61) or the total VFA production (P=0.70). It was concluded that germination processing of sorghum grain is affected by temperature conditions during active germination and not by temperature during steeping. In practice, the response of sorghum grain to germination processing will likely depend on seasonal temperature conditions and the variety of sorghum.

Fermentability and degradability of sorghum grain following soaking, aerobic or anaerobic treatment

This study determined the effect of soaking (steeping), aerobic (germination) and anaerobic (reconstitution) treatments on the solubility of carbohydrate and nitrogen, and on the in vitro fermentability and in sacco degradability of dry matter (DM) of sorghum grain ( Sorghum bicolor cv Western Red). Samples of grain were dry rolled (DR, control), soaked (SK), stored anaerobically for 21 d (reconstitution, AS21), stored aerobically for 5 d (germination, G5) or stored aerobically for 5 d and then anaerobically for 16 d (G5AS16). Compared to the DR treatment, only G5AS16 treatment reduced ( P < 0.05) starch content by 2% units. Free sugar and buffer-soluble nitrogen contents were higher ( P < 0.05) in G5 and G5AS16 sorghum (21 and 54 g kg −1 DM; 241 and 375 g kg −1 total nitrogen, respectively) than in DR sorghum (8 g kg −1 DM; 142 g kg −1 total nitrogen, respectively). Free sugar and buffer-soluble nitrogen contents were similar ( P > 0.05) for SK, AS21 and DR control sorghum. Following a 5-h incubation in buffered rumen liquor, G5 and G5AS21 treatments increased ( P < 0.05) the in vitro fermentability of sorghum compared to the DR control treatment. This was indicated by increases in gas production (121–215%), volatile fatty acid (VFA) production (98.6–210%) and the amount of starch fermented (22–47%), following a 5-h incubation with buffered rumen liquor. In sacco degradability of DM in sorghum after 48 h of incubation was higher for G5 and G5AS16 (0.44 and 0.45, respectively) treatment than for DR, SK and AS21 treatments (0.39, 0.36 and 0.34, respectively). SK and AS21 treatments did not increase the degradability of sorghum compared to the DR treatment. It was concluded that the germination process occurring during aerobic treatment improved the fermentability and degradability of sorghum compared to other treatments investigated in this study.

Biodegradation of tannic acid in an in vitro ruminal system

The microbial degradation of tannic acid by ruminal fluid of cattle which had no prior exposure to tannin-rich diets, was studied in an in vitro syringe system. Tannic acid caused an increase in the gas volume and a decrease in the ammonia concentration of the buffered rumen liquor during incubation for 24–72 h. The fermentation products from tannic acid were gallic acid, pyrogallol and resorcinol. This is the first report on the ability of ruminal fluid taken from cattle unexposed and unadapted to tanniniferous diets to degrade tannic acid into simpler products.

The use of enzymes to predict the digestibility of animal feeds

AbstractSix techniques were investigated to find a quick, reliable laboratory method for the prediction of the in‐vivo digestibility of animal feeds. Four of these techniques comprised two digestions, one with pepsin in hydrochloric acid followed by another with cellulase. In three cases the cellulase was derived from Trichoderma viride which was used at two concentrations; in addition, at one of these concentrations a step using amyloglucosidase was introduced. In another technique cellulase derived from Aspergillus niger was tested to determine how effectively it dealt with starchy materials. The fifth technique comprised the separation of the cell walls by digestion with neutral detergent solution followed by digestion with cellulase (derived from T. viride) with an additional step using amyloglucosidase for samples containing starch. The last method was an in‐vitro technique using buffered rumen liquor followed by pepsin in hydrochloric acid. Because this technique proved to estimate digestibility more accurately than any of the simple chemical methods it provided a guide to the efficiency of the enzyme techniques. The digestible organic matter in the dry matter (DOMD) was determined in each case and correlated against the DOMD value obtained using an in‐vivo technique. Two of the methods showed as good a prediction of DOMD as the in‐vitro method using rumen liquor, and the correlation coefficients were comparable. The repeatability of the methods calculated as the coefficient of variation varied between 2.5 and 3.7%. For roughages containing little or no starch the shorter techniques provided an adequate evaluation of DOMD. Starches in the cereals were adequately broken down only when the additional digestion using an amyloglucosidase (Agidex) was included; this is illustrated by the correlation values obtained.