Molar Proportions Of Volatile Fatty Acids Research Articles

Ruminal Microbiome Differences in Angus Steers with Differing Feed Efficiencies during the Feedlot Finishing Phase.

The catabolic activity of the ruminal microbial community of cattle enables the conversion of low-quality feedstuffs into meat and milk. The rate at which this conversion occurs is termed feed efficiency, which is of crucial importance given that feed expenses account for up to 70% of the cost of animal production. The present study assessed the relationship between cattle feed efficiency and the composition of their ruminal microbial communities during the feedlot finishing period. Angus steers (n = 65) were fed a feedlot finishing diet for 82 days and their growth performance metrics were evaluated. These included the dry matter intake (DMI), average daily gain (ADG), and residual feed intake (RFI). Steers were rank-ordered based upon their RFI, and the five lowest RFI (most efficient) and five highest RFI (least efficient) steers were selected for evaluations. Ruminal fluid samples were collected on days 0 and 82 of the finishing period. Volatile fatty acids (VFA) were quantified, and microbial DNA was extracted and the 16S rRNA gene was sequenced. The results showed that the ADG was not different (p = 0.82) between efficiency groups during the 82-day feedlot period; however, the efficient steers had lower (p = 0.03) DMI and RFI (p = 0.003). Less-efficient (high RFI) steers developed higher (p = 0.01) ruminal Methanobrevibacter relative abundances (p = 0.01) and tended (p = 0.09) to have more Methanosphaera. In high-efficiency steers (low RFI), the relative abundances of Ruminococcaceae increased (p = 0.04) over the 82-day period. The molar proportions of VFA were not different between the two efficiency groups, but some changes in the concentration of specific VFA were observed over time. The results indicated that the ruminal microbial populations of the less-efficient steers contained a greater relative abundance of methanogens compared to the high-efficiency steers during the feedlot phase, likely resulting in more energetic waste in the form or methane and less dietary energy being harvested by the less-efficient animals.

Exhalomics as a noninvasive method for assessing rumen fermentation in dairy cows: Can exhaled-breath metabolomics replace rumen sampling?

Previously, we used secondary electrospray ionization-mass spectrometry (SESI-MS) to investigate the diurnal patterns and signal intensities of exhaled (EX) volatile fatty acids (VFA) of dairy cows. The current study aimed to validate the potential of an exhalomics approach for evaluating rumen fermentation. The experiment was conducted in a switchback design, with 3 periods of 9 d each, including 7 d for adaptation and 2 d for sampling. Four rumen-cannulated original Swiss Brown (Braunvieh) cows were randomly assigned to 1 of 2 diet sequences (ABA or BAB): (A) low starch (LS; 6.31% starch on a dry matter basis) and (B) high starch (HS; 16.2% starch on a dry matter basis). Feeding was once per day at 0830 h. Exhalome (with the GreenFeed System), and rumen samples were collected 8 times to represent every 3 h of a day, and EX-VFA and ruminal (RM)-VFA were analyzed using SESI-MS and HPLC, respectively. Furthermore, the VFA concentration in the gas phase (HR-VFA) was predicted based on RM-VFA and Henry's Law (HR) constants. No interactions were identified between the types of diets (HS vs. LS) and the measurement methods on daily average VFA profiles (RM vs. EX or HR vs. EX), suggesting a consistent performance among the methods. Additionally, when the 3-h interval VFA data from HS and LS diets were analyzed separately, no interactions were observed between methods and time of day, indicating that the relative daily pattern of VFA molar proportions was similar regardless of the VFA measurement method used. The results revealed that the levels of acetate sharply increased immediately after feeding, trailed by an increase in the acetate:propionate ratio and a steady increase for propionate (2 h after feeding the HS diet, 4 h for LS), and butyrate. This change was more pronounced for the HS diet than the LS diet. However, there was no overall diet effect on the VFA molar proportions, although the measurement methods affected the molar proportions. Furthermore, we observed a strong positive correlation between the levels of RM and EX acetate for both diets (HS: r = 0.84; LS: r = 0.85), RM and EX propionate (r = 0.74), and RM and EX acetate:propionate ratio (r = 0.80). Both EX-VFA and RM-VFA exhibited similar responses to feeding and dietary treatments, suggesting that EX-VFA could serve as a useful proxy for characterizing RM-VFA molar proportions to evaluate rumen fermentation. Similar relationships were observed between RM-VFA and HR-VFA. In conclusion, this study underscores the potential of exhalomics as a reliable approach for assessing rumen fermentation. Moving forward, research should further explore the depth of exhalomics in ruminant studies to provide a comprehensive insight into rumen fermentation metabolites, especially across diverse dietary conditions.

Ensiling Characteristics, In Vitro Rumen Fermentation Patterns, Feed Degradability, and Methane and Ammonia Production of Berseem (Trifolium alexandrinum L.) Co-Ensiled with Artichoke Bracts (Cynara cardunculus L.).

This study investigated the effect of co-ensiling increasing levels of artichoke bracts (Cynara cardunculus L.) with berseem (Trifolium alexandrinum L.) (100:0, 75:25, 50:50, 25:75, and 0:100, respectively) on silage quality after 0, 30, 60, and 120 days. Moreover, the in vitro rumen fermentation characteristics and methane (CH4) and ammonia (NH3-N) production were evaluated using a buffalo inoculum source. The results showed that pH of the silage and the concentration of acetic, propionic, butyric acid, and NH3-N significantly decreased (L; p < 0.01) with the increasing amounts of artichoke bracts in the mixture. At 30 and 60 days of ensiling, the highest lactic acid concentration was observed at intermediate proportions of artichoke bracts (p < 0.01). Cumulative gas production was higher in artichoke bracts than in the berseem silage. After 24 h of incubation, the highest value (p < 0.05) of truly dry matter, organic matter, natural detergent fiber degradability, and NH3-N concentration was recorded with 500 g/kg of forage mixtures. As the artichoke bract concentration increased, the partitioning factor and ruminal pH declined linearly (p ≤ 0.05). No significant differences were observed for total volatile fatty acids and volatile fatty acids molar proportions. In summary, co-ensiling artichoke bracts with berseem at a ratio of 1:1 might be a promising and easy method for the production of high-quality silage from legume forage with positively manipulating rumen fermentation.

Effects of Cashew Nut Shell Extract on Ruminal Fermentation and Nutrient Digestibility under Continuous Culture

The overall objective of this study was to determine the dose response to four levels of cashew nut shell extract in a granulated form (CNSE, containing 59% anacardic acid and 18% cardol) on culture pH, rumen fermentation metabolites, and apparent nutrient digestibility in continuous culture fermenters. The study was conducted as a generalized randomized complete block design with four treatments and four replications per treatment. The four treatments were randomly assigned to eight fermenters for two incubation runs of 10 d. Treatments consisted of (1) Control (CO, no CNSE), (2) Control plus 100 ppm of CNSE, (3) Control plus 200 ppm of CNSE, and (4) Control plus 300 ppm of CNSE. Fermenters were fed 52 g/d (DM basis) of a total mixed ration (TMR; 17.0% crude protein (CP), 29.7% NDF, and 29.9% starch), divided between two feedings at 0800 and 2000 h. The apparent digestibility of dry matter (DM), organic matter (OM), and neutral detergent fiber (NDF) were not affected by CNSE supplementation. Similarly, CNSE had no effect on culture pH, total volatile fatty acids (VFA) or individual VFA molar proportions. These results suggest that at the dosages evaluated in this study, CNSE has no impact on the rumen fermentation profile and the apparent nutrient digestibility under continuous culture conditions.

Effect of 3-nitrooxypropanol on enteric methane emissions of feedlot cattle fed a tempered barley-based diet with canola oil.

A dose-response experiment was designed to examine the effect of 3-nitrooxypropanol (3-NOP) on methane (CH4) emissions, rumen function and performance of feedlot cattle fed a tempered barley-based diet with canola oil. Twenty Angus steers of initial body weight (BW) of 356 ± 14.4kg were allocated in a randomized complete block design. Initial BW was used as the blocking criterion. Cattle were housed in individual indoor pens for 112 d, including the first 21 d of adaptation followed by a 90-d finishing period when five different 3-NOP inclusion rates were compared: 0mg/kg DM (control), 50mg/kg DM, 75mg/kg DM, 100mg/kg DM and 125mg/kg DM. Daily CH4 production was measured on d 7 (last day of starter diet), 14 (last day of the first intermediate diet), and 21 (last day of the second intermediate diet) of the adaptation period and on d 28, 49, 70, 91 and 112 of the finisher period using open circuit respiration chambers. Rumen digesta samples were collected from each steer on the day prior to chamber measurement post-feeding, and pre-feeding on the day after the chamber measurement, for determination of rumen volatile fatty acids (VFA), ammonium-N, protozoa enumeration, pH and reduction potential. Dry matter intake (DMI) was recorded daily and BW weekly. Data were analyzed in a mixed model including period, 3-NOP dose and their interaction as fixed effects, and block as a random effect. Our results demonstrated both a linear and quadratic (decreasing rate of change) effect on CH4 production (g/d) and CH4 yield (g/kg DMI) as 3-NOP dose increased (P < 0.01). The achieved mitigation for CH4 yield in our study ranged from approximately 65.5% up to 87.6% relative to control steers fed a finishing feedlot diet. Our results revealed that 3-NOP dose did not alter rumen fermentation parameters such as ammonium-N, volatile fatty acids (VFA) concentration nor VFA molar proportions. Although this experimental design was not focused on the effect of 3-NOP dose on feedlot performance, no negative effects of any 3-NOP dose were detected on animal production parameters. Ultimately, the knowledge on the CH4 suppression pattern of 3-NOP may facilitate sustainable pathways for the feedlot industry to lower its carbon footprint.

PSVI-19 Effect of Processing Methods and Inclusion Level of Dried raw Garlic on in Vitro Fermentation Profile and Methane Production in a Corn Silage-Based Diet

Abstract Garlic (Allium sativum) contains secondary compounds known to modify rumen fermentation parameters and decrease methane (CH4) emissions; however, results from previous research on the effects of garlic-byproduct supplementation are quite variable. The objective of this experiment was to evaluate the effects of increasing inclusion levels and processing methods of dried raw garlic on in vitro ruminal fermentation and CH4 production. The experiment was designed as a randomized complete block design, where 24 h incubations were conducted on three separate days, with corn silage and gin trash (80:20, respectively) used as basal substrate. Treatments were arranged in a 3 × 3 factorial with garlic inclusion (0, 2.5, and 5% of the diet) and processing method (freeze-dried, oven-dried, and autoclaved) as factors. Three ruminally cannulated steers were fed ad libitum a corn silage and gin trash diet, for at least 35 d prior to collection of ruminal fluid used as inoculum. Final pH, concentration of volatile fatty acids (VFA) and ammonia nitrogen (NH3-N), in vitro organic matter digestibility (IVOMD), total gas production, and CH4 concentrations were determined at the end of incubation. Data were analyzed using the Mixed procedure of SAS. Final pH and concentration of total VFA were not affected (P &amp;gt; 0.05) by the processing method nor inclusion level of garlic. Increasing garlic inclusion decreased NH3-N concentration (P &amp;lt; 0.05), reduced (P &amp;lt; 0.05) the proportion of acetate, tended to increase (P &amp;lt; 0.10) the proportion of propionate, and tended to decrease (P &amp;lt; 0.10) the acetate:propionate ratio. Neither garlic inclusion level nor processing methods modified IVOMD, total gas production, or CH4 concentration. Modifications in VFA molar proportions and NH3-N concentration may be related to differences in N concentration in the incubated substrates. In conclusion, neither processing method nor garlic level inclusion modified CH4 production or nutrient digestibility.

Effect of incremental proportions of Desmanthus spp. in isonitrogenous forage diets on growth performance, rumen fermentation and plasma metabolites of pen-fed growing Brahman, Charbray and Droughtmaster crossbred beef steers.

Desmanthus (Desmanthus spp.), a tropically adapted pasture legume, is highly productive and has the potential to reduce methane emissions in beef cattle. However, liveweight gain response to desmanthus supplementation has been inconclusive in ruminants. This study aimed to evaluate weight gain, rumen fermentation and plasma metabolites of Australian tropical beef cattle in response to supplementation with incremental levels of desmanthus forage legume in isonitrogenous diets. Forty-eight Brahman, Charbray and Droughtmaster crossbred beef steers were pen-housed and fed a basal diet of Rhodes grass (Chloris gayana) hay supplemented with 0, 15, 30 or 45% freshly chopped desmanthus forage on dry matter basis, for 140 days. Varying levels of lucerne (Medicago sativa) hay were added in the 0, 15 and 30% diets to ensure that all diets were isonitrogenous with the 45% desmanthus diet. Data were analyzed using the Mixed Model procedures of SAS software. Results showed that the proportion of desmanthus in the diet had no significant effect on steer liveweight, rumen volatile fatty acids molar proportions and plasma metabolites (P ≥ 0.067). Total bilirubin ranged between 3.0 and 3.6 μmol/L for all the diet treatments (P = 0.67). All plasma metabolites measured were within the expected normal range reported for beef cattle. Rumen ammonia nitrogen content was above the 10 mg/dl threshold required to maintain effective rumen microbial activity and maximize voluntary feed intake in cattle fed low-quality tropical forages. The average daily weight gains averaged 0.5 to 0.6 kg/day (P = 0.13) and were within the range required to meet the target slaughter weight for prime beef markets within 2.5 years of age. These results indicate that desmanthus alone or mixed with other high-quality legume forages can be used to supplement grass-based diets to improve tropical beef cattle production in northern Australia with no adverse effect on cattle health.

Method of diet delivery to dairy cows: Effects on nutrient digestion, rumen fermentation, methane emissions from enteric fermentation and stored manure, nitrogen excretion, and milk production

This study was conducted to examine the effect of method of diet delivery to dairy cows on enteric CH4 emission, milk production, rumen fermentation, nutrient digestion, N excretion, and manure CH4 production potential. Sixteen lactating cows were used in a crossover design (35-d period) and fed ad libitum twice daily a diet [52:48, forage:concentrate ratio; dry matter (DM) basis] provided as forages and concentrates separately (CF) or as a total mixed ration (TMR). For the CF treatment, concentrates were offered first followed by mixed forages 45 min afterward. Method of diet delivery had no effect on DM intake, but neutral detergent fiber (NDF) intake was greater when the diet was delivered as TMR as compared with CF. Apparent total-tract digestibility of DM, crude protein, and gross energy was slightly (1 percentage unit) lower when the diet was offered as TMR than when offered as CF. In contrast, NDF digestibility was greater when the cows were fed TMR versus CF. Although average daily ruminal pH was not affected by method of diet delivery, daily duration of ruminal pH <5.6 was less when the diet was delivered as TMR as compared with CF (0.9 h/d versus 3.7 h/d). Delivering the diet as TMR increased ruminal total volatile fatty acid and NH3 concentrations, but had no effect on acetate, propionate, or branched-chain volatile fatty acid molar proportions. Yields of milk, milk fat, or milk protein, and milk production efficiency (kg of milk/kg of DM intake or g of N milk/g of N intake) were not affected by the method of diet delivery. Daily production (g/d), yield (% gross energy intake), and emission intensity (g/kg of energy-corrected milk) of enteric CH4 averaged 420 g/d, 4.9%, and 9.6 g/kg and were not affected by diet delivery method. Fecal N output was greater when the diet was delivered as TMR versus CF, whereas urinary N excretion (g/d, % N intake) was not affected. Manure volatile solids excretion and maximal CH4 production potential were not affected by method of diet delivery. Under the conditions of this study, delivering the diet as concentrates and forages separately versus a total mixed ration had no effect on milk production, enteric CH4 energy losses, urinary N, or maximal manure CH4 emission potential. However, feeding the diet as total mixed ration compared with feeding concentrates and forages separately attenuated the extent of postprandial decrease in ruminal pH, which has contributed to improving NDF digestibility.

Rumen volatile fatty acid molar proportions, rumen epithelial gene expression, and blood metabolite concentration responses to ruminally degradable starch and fiber supplies

The objective of this work was to characterize rumen volatile fatty acid (VFA) concentrations, rumen epithelial gene expression, and blood metabolite responses to diets with different starch and fiber sources. Six ruminally cannulated yearling Holstein heifers (body weight = 330 ± 11.3 kg) were arranged in a partially replicated Latin square experiment with 4 treatments consisting of different starch [barley (BAR) or corn (CRN)] and fiber [timothy hay (TH) or beet pulp (BP)] sources. Treatments were arranged as a 2 × 2 factorial. Beet pulp and TH were used to create relative changes in apparent ruminal fiber disappearance, whereas CRN and BAR were used to create relative changes in apparent ruminal starch disappearance. Each period consisted of 3 d of diet adaptation and 15 d of dietary treatment. In situ disappearance of fiber and starch were estimated from bags incubated in the rumen from d 10 to 14. From d 15 to 17, rumen fluid was collected every hour from 0500 to 2300 h. Rumen fluid samples were pooled by animal/period and analyzed for pH and VFA concentrations. On d 18, 60 to 80 papillae were biopsied from the epithelium and preserved for gene expression analysis. On d 18, one blood sample per heifer was collected from the coccygeal vessel. In situ ruminal starch disappearance rate (7.30 to 8.72%/h for BAR vs. 7.61 to 10.5%/h for CRN) and the extent of fiber disappearance (22.2 to 33.4% of DM for TH vs. 34.4 to 38.7% of DM for BP) were affected by starch and fiber source, respectively. Analysis of VFA molar proportions showed a shift from propionate to acetate, and valerate to isovalerate on TH diets compared with BP. Corn diets favored propionate over butyrate in comparison to BAR diets. Corn diets also had higher molar proportions of valerate. Expression of 1 gene (SLC9A3) were increased in BP diets and 2 genes (BDH1 and SLC16A4) tended to be increased in TH diets. Plasma acetate demonstrated a tendency for a starch by fiber interaction with BAR-BP diets having the highest plasma acetate, but other metabolites measured were not significant. These results suggest that TH has the greatest effect on shifts in VFA molar proportions and epithelial transporters, but does not demonstrate shifts in blood metabolite concentrations.

Evaluation of the Nutrient Composition, In Vitro Fermentation Characteristics, and In Situ Degradability of Amaranthus caudatus, Amaranthus cruentus, and Amaranthus hypochondriacus in Cattle.

Simple SummaryThe amaranth plants, one of the crops that can grow in poor soil and areas with extreme weather conditions—high temperature and limited rainfall—showed high potential feed value as forage for ruminants. An extensive study will help extend its utilization as an alternative protein and fiber feed source in cattle feeding. In this study, the nutrient compositions of the three different species of amaranth, Amaranthus caudatus, A. cruentus, and A. hypochondriacus, were evaluated. Rumen fluid was incubated with the amaranth plants to evaluate fermentation characteristics (total gas production, total volatile fatty acids concentration, pH, and ammonia concentration). There were no differences among the different amaranth species, but all amaranth showed favorable fermentation values. The effective degradabilities of dry matter and crude protein of the amaranth forages were also determined. Compared to other studies, the effective degradabilities of dry matter (33–56%) and crude protein (27–59%) of the amaranth were lower; these results could be due to the maturity stage at which the forages were harvested. In terms of chemical composition, the amaranth forages showed better nutritive value than the locally produced forages in Chungcheong province of Korea. The amaranth forages showed 11.95–14.19% crude protein (CP), 45.53–70.88% neutral detergent fiber (NDF), and 34.17–49.83% acid detergent fiber (ADF) contents. The high nutrient composition, highly effective degradability of dry matter, and crude protein coupled with the favorable fermentation characteristics suggest that the amaranth forages showed good to excellent feed quality to cattle.The amaranth plants showed high potential feed value as forage for ruminants. An in-depth study of this plant, particularly in cattle, will help extend its utilization as an alternative protein and fiber feed source in cattle feeding. In this study, the nutrient compositions of three different species of amaranth, Amaranthus caudatus L., Amaranthus cruentus L., and Amaranthus hypochondriacus L.—two varieties for each species, A.ca 74, A.ca 91, A.cu 62, A.cu 66, A. hy 30, and A. hy 48—were evaluated. The in vitro technique was used to evaluate the fermentation characteristics such as total gas production, total volatile fatty acids (VFA) concentration, pH, and ammonia concentration of the rumen fluid. Moreover, the effective degradabilities of dry matter (EDDM) and crude protein (EDCP) of the amaranth forages were determined through in situ bag technique. The amaranth forages: A. caudatus, A. cruentus, and A. hypochondriacus showed better nutritive value than the locally produced forages in Chungcheong province of Korea. The CP of the amaranth ranged from 11.95% to 14.19%, and the neutral detergent fiber (NDF) and acid detergent fiber (ADF) contents ranged from 45.53% to 70.88% and 34.17% to 49.83%, respectively. Among the amaranth varieties, A. hypochondriacus 48 showed the most excellent ruminant feed nutrient quality (CP, 14.19%; NDF, 45.53%; and ADF, 34.17%). The effective degradabilities of dry matter (EDDM; 33–56%) and crude protein EDCP (27–59%) of the amaranth were lower compared to other studies, which could be due to the maturity stage at which the forages were harvested. Nonetheless, A. hypochondriacus 48 showed the highest EDDM (56.73%) and EDCP (59.09%). The different amaranth species did not differ greatly in terms of total VFA concentration or molar proportions, total gas production, or ammonia-N concentration. The high nutrient composition, and highly effective degradability of dry matter and crude protein, coupled with the favorable fermentation characteristics, suggest that the amaranth forages showed good to excellent feed quality for cattle.

Comparison of Two Sampling Techniques for Evaluating Ruminal Fermentation and Microbiota in the Planktonic Phase of Rumen Digesta in Dairy Cows

The objective of this experiment was to compare ruminal fluid samples collected through rumen cannula (RC) or using an oral stomach tube (ST) for measurement of ruminal fermentation and microbiota variables. Six ruminally cannulated lactating Holstein cows fed a standard diet were used in the study. Rumen samples were collected at 0, 2, 4, 6, 8, and 12 h after the morning feeding on two consecutive days using both RC and ST techniques. Samples were filtered through two layers of cheesecloth and the filtered ruminal fluid was used for further analysis. Compared with RC, ST samples had 7% greater pH; however, the pattern in pH change after feeding was similar between sampling methods. Total volatile fatty acids (VFA), acetate and propionate concentrations in ruminal fluid were on average 23% lower for ST compared with RC. There were no differences between RC and ST in VFA molar proportions (except for isobutyrate), ammonia and dissolved hydrogen (dH2) concentrations, or total protozoa counts, and there were no interactions between sampling technique and time of sampling. Bacterial ASV richness was higher in ST compared with RC samples; however, no differences were observed for Shannon diversity. Based on Permanova analysis, bacterial community composition was influenced by sampling method and there was an interaction between sampling method and time of sampling. A core microbiota comprised of Prevotella, S24-7, unclassified Bacteroidales and unclassified Clostridiales, Butyrivibrio, unclassified Lachnospiraceae, unclassified Ruminococcaceae, Ruminococcus, and Sharpea was present in both ST and RC samples, although their relative abundance varied and was influenced by an interaction between sampling time and sampling method. Overall, our results suggest that ruminal fluid samples collected using ST (at 180 to 200 cm depth) are not representative of rumen pH, absolute values of VFA concentrations, or bacterial communities >2 h post-feeding when compared to samples of ruminal fluid collected using RC. However, ST can be a feasible sampling technique if the purpose is to study molar proportions of VFA, protozoa counts, dH2, and ammonia concentrations.

PSVII-10 Evaluation of different biochar sources added at two inclusion levels in a grass hay- based diet on dry matter disappearance and ruminal fermentation parameters in vitro

Abstract This study evaluated the impact of adding biochar to a grass hay-based diet on in vitro dry matter disappearance (DMD), total gas production (GP), methane production (CH4), volatile fatty acid (VFA) and ammonia nitrogen (NH3-N) concentration. Treatments were arranged in a factorial design with 7 biochar products differing in origin (pine or coconut-based) and physical properties (course, fine or loose spheres) at 2 inclusion levels (2.25% and 4.5% of DM) and a grass hay-only control. On each three successive weeks, rumen fluid was collected from 2 ruminally cannulated beef heifers fed a grass hay-based diet, pooled and mixed with a buffer solution (1:2) and treatments incubated in triplicate for 48 h. Gas pressure was recorded at 3, 6, 9, 12, 18, 24, 36 and 48 h of incubation, and gas was collected at each time to measure CH4. At 48 h, pH was measured and the supernatant sampled for determination of VFA and NH3-N, while the residue was used to estimate DMD. Dry matter disappearance decreased (P &amp;lt; 0.05) with increasing addition of biochar. Biochar did not affect GP or CH4, but did reduce (P &amp;lt; 0.05) gas production per g of DM incubated (111.7 vs 107.8 ml/g), but not per g DMD. The lower GP per g DM incubated could be related to the increased proportion of the diet as biochar, without it contributing to GP. Biochar, irrespective of level did not affect the total or molar proportions of VFA. There was a biochar x inclusion level interaction (P &amp;lt; 0.04) in NH3-N concentration. Biochar type and inclusion level in the present study had little or no significant effect on in vitro fermentation and did not mitigate CH4 production from a grass hay-based diet.

Pre-Caecal Disappearance of Starch and Volatile Fatty Acid (VFA) Content in Digesta of Caecum of Growing Pigs Fed with Ripe Hulled Shredded Acorns in Their Diet

Physical properties and chemical composition of caecum chyme, with particular regard to starch pre-caecal disappearance and volatile fatty acid (VFA, C2-C5) content, were determined in 16 growing pigs fed raw acorns in the diet. Pigs were singly housed and divided into two experimental feeding groups (each n = 8). The control group was fed a conventional complete diet for pigs (pelleted; 890 g dry matter (DM)/kg, 435 g starch/kg DM, based on cereals and soybean extraction meal). The acorn-fed group was offered a combined diet, 70% of ripe whole shredded acorns (695 g DM/kg, 447 g starch/kg DM) and 30% of the control diet. After one week, all 16 animals were slaughtered, and contents of the caudal ileum and caecum were analyzed for dry matter, crude ash, and starch. Fresh caecum chyme was analyzed for VFA content and molar proportion of acetic, propionic, n-butyric, n-valeric acid were determined. VFA per kg of organic matter (OM) in the caecum differed significantly between groups (p = 0.047). Pre-caecal disappearance (pCD) of acorn native starch turned out to be high (&gt;95%). Estimated VFA content in caecum in the acorn combined diet resulted in 8.72 g per kg of OM, and the relationship based on molar proportion between acetic:propionic:n-butyric:n-valeric acid was 1:0.2:0.1:0.04, which pointed to higher proportions of n-valeric acid when hulled shredded acorns are fed to pigs.

Effects of fertiliser nitrogen rate to spring grass on apparent digestibility, nitrogen balance, ruminal fermentation and microbial nitrogen production in beef cattle and in vitro rumen fermentation and methane output

The effects of two fertiliser nitrogen (N) application rates - 15 (LN) or 80 (HN) kg N/ha - to Lolium perenne dominant swards in spring, on grass dry matter (DM) intake, digestion, rumen fermentation, microbial N production and N-balance in beef cattle, and in vitro fermentation and methane production were studied. Sixteen Charolais steers with a mean live weight (s.d.) of 475 (18.4) kg, were used in a completely randomised block design experiment and offered zero-grazed grass harvested 21-d post N application. The same grass was incubated in an eight-vessel RUSITEC in a completely randomised block design experiment. The HN treatment had a 540 kg/ha higher grass DM yield, and a 20 g/kg DM higher crude protein (CP) concentration compared to LN. There was no difference (P > 0.05) in DM intake, or in vivo DM, organic matter (OM) and N digestibility between treatments. Rumen fermentation variables pH, lactic acid, ammonia (NH3) and total volatile fatty acid (VFA) concentration were similar (P > 0.05) for both treatments. Nitrogen intake was 19 g/d higher (P < 0.05) for HN compared to LN. Total and urine N loss was 16 and 14 g/d greater (P < 0.05), respectively, for HN compared to LN, but faecal N loss did not differ (P > 0.05) between treatments. The quantity of N retained and N-use efficiency did not differ (P > 0.05) between LN and HN. Plasma urea concentration was 1 mmol/L greater (P < 0.05) for HN compared to LN. Estimated microbial N production was greater (P < 0.05) for HN compared to LN. In vitro NH3 concentrations were higher (P < 0.05) for HN compared to LN, whereas in vitro rumen pH, lactic acid and VFA concentrations and molar proportions did not differ (P > 0.05) between HN and LN. In vitro methane and total gas output were not different (P > 0.05) between treatments. Reducing fertiliser N application rate to grass in spring reduced total and urinary N excretion, which has environmental benefits, with no effects on in vitro methane output.

Effects of long-term diet supplementation with Gliricidia sepium foliage mixed with Enterolobium cyclocarpum pods on enteric methane, apparent digestibility, and rumen microbial population in crossbred heifers1.

In the last decades, strategies have been evaluated to reduce rumen methane (CH4) production by supplementing tropical forages rich in secondary compounds; however, most of these beneficial effects need to be validated in terms of their persistence over time. The aim of this study was to assess CH4 emissions over time in heifers fed with and without Gliricidia sepium foliage (G) mixed with ground pods of Enterolobium cyclocarpum(E). Two groups of 4 crossbred (Bos taurus x Bos indicus) heifers (284 ±17 kg initial weight) were fed with 2 diets (0% and 15% of a mixture of the pods and foliage [E + G:0 and E + G:15, respectively]) over 80 d, plus 2 wk before the experiment, in which every animal was fed a legume and pod-free diet. Every 14 d, CH4 production, apparent digestibility, volatile fatty acids (VFA), and microbial population were quantified for each animal. The experiment was conducted with a repeated measurements design over time. Diets fed differed in terms of their crude protein (CP), condensed tannins, and saponins content supplied by E. cyclocarpum and G. sepium. For most of the experiment, dry matter intake (DMI) and digestible dry-matter intake (DDMI) were 6.3 kg DMI/d and 512 g DDMI/kg, respectively, for both diets (diet: P > 0.05). Apparent digestible crude protein (DCP) was reduced by 21 g DCP/kg DM when the diet was supplemented with E + G:15 (P = 0.040). Molar proportions of VFA's in the rumen did not differ between diets or in time (P > 0.05). Daily methane production, expressed in relation to DMI, was 23.95 vs. 23.32 g CH4/kg DMI for the diet E + G:0 and E + G:15, respectively (diet: P = 0.016; Time: P > 0.05). Percent gross energy loss as CH4 (Ym) with grass-only diets was above 8.1%, whereas when feeding heifers with the alternate supplementation, Ym values of 7.59% (P = 0.016) were observed. The relative abundance of total bacterial, protozoa, and methanogenic archaeal replicates was not affected by time nor by the incorporation of legume and pods into the diet (P > 0.05). Results suggest that addition of G. sepium mixed with E. cyclocarpum pods can reduce CH4 production in heifers and this response remains over time, without effect on microbial population and VFA concentration and a slight reduction in CPD digestibility.

In vitro fermentation of browse species using goat rumen fluid in relation to browse polyphenol content and composition

The effect of browse species tannins (using polyethylene glycol, PEG 6000) on in vitro gas production (GP), methane (CH4), ammonia (NH3), volatile fatty acids (VFA) and in vitro organic matter digestibility (IVOMD) were studied. Approximately 0.5g of air-dried A. etbaica, C. farinosa, C. tomentosa, D. angustifolia, D. cinerea, E. racemosa, M. angolensis, M. senegalensis, R. natalensis and S. singueana leaves were used as substrates in an automated in vitro system. Proanthocyanidin (PA) contents were quantified using the modified HCl-butanol method and the PA composition analysed by ultra performance liquid chromatography using a diode array detector coupled with electrospray ionization mass spectrometry (UPLC-DAD-ESI–MS/MS). Substrates were inoculated for 72h in buffered rumen fluid (2:1, v/v), pooled from three goats fed a grass silage and concentrate diet, on three separate occasions. Each substrate was incubated in triplicate with or without inclusion of PEG 6000 and GP measured. During incubation, head space gas samples were taken at 0, 3, 6, 9, 12, 24, 30, 48, 54, and 72h and analysed for CH4. Volatile fatty acids, NH3 and IVOMD were determined after 72h of incubation. Data from three runs were averaged for analysis. Addition of PEG increased (P<0.0001) GP, CH4, NH3 and total VFA indicating that PA were mainly involved in reducing methanogenesis but also digestibility. Prodelphinidins were found to be the major PA affecting fermentation. Also the contribution of quercetin, myricetin and kaempferol derivatives in CH4 reduction were evident. Changes in the molar proportions of VFA with PEG addition indicated that PA affected fermentation pathways. The absence of PEG effect on IVOMD was due to artefacts from the tannin-PEG complexes interfering with the incubation residue measurement. Overall, the effect of tannin-containing browse on in vitro fermentation characteristics was mainly due to PA with the possible minor effects of other phenolic and non-phenolic compounds.

Influence of zeolite (clinoptilolite) supplementation on characteristics of digestion and ruminal fermentation of steers fed a steam-flaked corn-based finishing diet

Four Holstein steers (328 ± 14 kg) with ruminal and duodenal cannulas were used in a 4 × 4 Latin square design to examine the effect of level of zeolite supplementation (0, 10, 20 or 30 g clinoptilolite-Ca/kg of diet) in a steam-flaked corn-based finishing diet on the characteristics of ruminal fermentation and nutrient digestion. Zeolite was top-dressed with the basal diet at time of feeding; therefore, intake of organic matter (OM), neutral detergent fibre (NDF), starch and N was equal in all treatments. Zeolite supplementation did not affect site and extent of digestion of N or NDF. Zeolite linearly decreased (P &lt; 0.01) NH3-N flow to duodenum, but did not affect duodenal flow of microbial N, feed N, or microbial efficiency. Inclusion of zeolite linearly increased (P = 0.02) ruminal OM digestion and tended (linear, P = 0.08) to increase ruminal digestion of starch. Zeolite supplementation increased (linear effect, P &lt; 0.01) faecal excretion of ash, resulting in decreased total tract DM digestion (linear effect, P = 0.04). However, total tract digestion of OM tended to increase (P = 0.06) with increasing zeolite supplementation. Thus, dilution of the diet with zeolite did not depress dietary digestible energy. There were no treatment effects on ruminal pH. Total ruminal volatile fatty acid concentration tended (P = 0.08) to increase as zeolite increased. Zeolite supplementation decreased ruminal molar proportion of acetate and increased molar proportion of propionate (linear effect, P &lt; 0.01), resulting in decreased (linear effect, P &lt; 0.01) of acetate: propionate ratio. Supplementation with zeolite at levels of up to 30 g of zeolite/kg of diet may enhance ruminal OM digestion, largely due to the tendency of increased ruminal starch digestion. Zeolite supplementation altered ruminal volatile fatty acid molar proportions (decreased acetate : propionate ratio), and therefore may have an positive impact on energy efficiency.

Changes in in vitro gas and methane production from rumen fluid from dairy cows during adaptation to feed additives in vivo

The adaptation of dairy cows to methane (CH4)-mitigating feed additives was evaluated using the in vitro gas production (GP) technique. Nine rumen-fistulated lactating Holstein cows were grouped into three blocks and within blocks randomly assigned to one of three experimental diets: Control (CON; no feed additive), Agolin RuminantR (AR; 0.05 g/kg dry matter (DM)) or lauric acid (LA; 30 g/kg DM). Total mixed rations composed of maize silage, grass silage and concentrate were fed in a 40 : 30 : 30 ratio on DM basis. Rumen fluid was collected from each cow at days −4, 1, 4, 8, 15 and 22 relative to the introduction of the additives in the diets. On each of these days, a 48-h GP experiment was performed in which rumen fluid from each individual donor cow was incubated with each of the three substrates that reflected the treatment diets offered to the cows. DM intake was on average 19.8, 20.1 and 16.2 kg/day with an average fat- and protein-corrected milk production of 30.7, 31.7 and 26.2 kg/day with diet CON, AR and LA, respectively. In general, feed additives in the donor cow diet had a larger effect on gas and CH4 production than the same additives in the incubation substrate. Incubation substrate affected asymptotic GP, half-time of asymptotic CH4 production, total volatile fatty acid (VFA) concentration, molar proportions of propionate and butyrate and degradation of organic matter (OMD), but did not affect CH4 production. No substrate×day interactions were observed. A significant diet×day interaction was observed for in vitro gas and CH4 production, total VFA concentration, molar proportions of VFA and OMD. From day 4 onwards, the LA diet persistently reduced gas and CH4 production, total VFA concentration, acetate molar proportion and OMD, and increased propionate molar proportion. In vitro CH4 production was reduced by the AR diet on day 8, but not on days 15 and 22. In line with these findings, the molar proportion of propionate in fermentation fluid was greater, and that of acetate smaller, for the AR diet than for the CON diet on day 8, but not on days 15 and 22. Overall, the data indicate a short-term effect of AR on CH4 production, whereas the CH4-mitigating effect of LA persisted.

Effect of forage to concentrate ratio with sorghum silage as a source of forage on rumen fermentation, N balance, and purine derivative excretion in limit-fed dairy heifers

Sorghum silage has been shown to be a good alternative to corn silage for dairy cows; however, studies regarding heifers are insufficiently explored. Therefore, the objective of this study was to evaluate effects of changing forage to concentrate ratio (FOR:CON) in diets based on sorghum silage on N digestibility, rumen fermentation, N balance, C excretion, and microbial N yield in limit-fed dairy heifers. A split-plot 4×4 Latin square design with 19-d periods (15 d of adaptation and 4 d of sampling) was conducted with 8 rumen cannulated dairy heifers (age 13.7±0.6mo and weight 364.8±17.6kg). Heifers were fed sorghum silage–based diets with 4 FOR:CON (85:15, 75:25, 65:35, and 55:45) balanced for similar metabolizable energy intake per unit of body weight and crude protein concentration. Diets were fed to allow 900 to 1,000g/d body weight gain and were fed once daily. Total collection of feces and urine was completed on d 15 to 19 to determine N, C, urea N, allantoin, uric acid, and creatinine excretion. Rumen contents were sampled on d 19 at 0, 1.5, 3, 4.5, 6, 9, 13, 17, 21, and 23h after feeding to measure pH, volatile fatty acid (VFA), ammonia-N, and free AA concentrations. The pH decreased linearly while ammonia-N and free AA levels increased linearly with decreasing FOR:CON of diets. Although mean total VFA did not differ among treatment diets, molar proportions of VFA did. Acetate proportion decreased while propionate and butyrate increased with decreasing FOR:CON. Intake of N and urea N excretion decreased with decreasing forage proportion in diets while total N excretion, apparent N digestibility, and N retention were not different. Intake of C and excretion in feces (g/d) decreased linearly with decreasing FOR:CON in diets. Creatinine, allantoin, and uric acid excretion were not affected by FOR:CON; however, microbial N yield tended to increase linearly with greater concentrate in diets. Heifers limit fed diets based on sorghum silage demonstrated the effect of available ammonia-N and readily fermentable carbohydrates with subsequent effects on nutrient utilization when different FOR:CON were applied. Based on the presented results, FOR:CON 65:35 had the most suitable balance of available ammonia-N and readily fermentable carbohydrates for the most optimal results.

Influence of periparturient and postpartum diets on rumen methanogen communities in three breeds of primiparous dairy cows.

BackgroundEnteric methane from rumen methanogens is responsible for 25.9 % of total methane emissions in the United States. Rumen methanogens also contribute to decreased animal feed efficiency. For methane mitigation strategies to be successful, it is important to establish which factors influence the rumen methanogen community and rumen volatile fatty acids (VFA). In the present study, we used next-generation sequencing to determine if dairy breed and/or days in milk (DIM) (high-fiber periparturient versus high-starch postpartum diets) affect the rumen environment and methanogen community of primiparous Holstein, Jersey, and Holstein-Jersey crossbreeds.ResultsWhen the 16S rRNA gene sequences were processed and assigned to operational taxonomic units (OTU), a core methanogen community was identified, consisting of Methanobrevibacter (Mbr.) smithii, Mbr. thaueri, Mbr. ruminantium, and Mbr. millerae. The 16S rRNA gene sequence reads clustered at 3 DIM, but not by breed. At 3 DIM, the mean % abundance of Mbr. thaueri was lower in Jerseys (26.9 %) and higher in Holsteins (30.7 %) and Holstein-Jersey crossbreeds (30.3 %) (P < 0.001). The molar concentrations of total VFA were higher at 3 DIM than at 93, 183, and 273 DIM, whereas the molar proportions of propionate were increased at 3 and 93 DIM, relative to 183 and 273 DIM. Rumen methanogen densities, distributions of the Mbr. species, and VFA molar proportions did not differ by breed.ConclusionsThe data from the present study suggest that a core methanogen community is present among dairy breeds, through out a lactation. Furthermore, the methanogen communities were more influenced by DIM and the breed by DIM interactions than breed differences.