Effects of bacterial direct-fed microbials on ruminal characteristics, methane emission, and milk fatty acid composition in cows fed high- or low-starch diets.

This study investigated the effects of bacterial direct-fed microbials (DFM) on ruminal fermentation and microbial characteristics, methane (CH4) emission, diet digestibility, and milk fatty acid (FA) composition in dairy cows fed diets formulated to induce different ruminal volatile fatty acid (VFA) profiles. Eight ruminally cannulated dairy cows were divided into 2 groups based on parity, days in milk, milk production, and body weight. Cows in each group were fed either a high-starch (38%, HS) or a low-starch (2%, LS) diet in a 55:45 forage-to-concentrate ratio on a dry matter (DM) basis. For each diet, cows were randomly assigned to 1 of 4 treatments in a Latin square design of (1) control (CON); (2) Propionibacterium P63 (P63); (3) P63 plus Lactobacillus plantarum 115 (P63+Lp); (4) P63 plus Lactobacillus rhamnosus 32 (P63+Lr). Strains of DFM were administered at 1010 cfu/d. Methane emission (using the sulfur hexafluoride tracer technique), total-tract digestibility, dry matter intake, and milk production and composition were quantified in wk 3. Ruminal fermentation and microbial characteristics were measured in wk 4. Data were analyzed using the mixed procedure of SAS (SAS Institute Inc., Cary, NC). The 2 diets induced different ruminal VFA profiles, with a greater proportion of propionate at the expense of acetate and butyrate for the HS diet. Greater concentrations of total bacteria and selected bacterial species of methanogenic Archaea were reported for the HS diet, whereas the protozoa concentration in HS decreased. For both diets, bacterial DFM supplementation raised ruminal pH (+0.18 pH units, on average) compared with CON. Irrespective of diet, P63+Lp and P63+Lr increased ruminal cellulase activity (3.8-fold, on average) compared with CON, but this effect was not associated with variations in ruminal microbial numbers. Irrespective of diet, no effect of bacterial DFM on ruminal VFA was observed. For the LS diet, supplementing cows with P63+Lr tended to decrease CH4 emission (26.5%, on average, when expressed per kilogram of milk or 4% fat-corrected milk). Only P63 supplementation to cows fed the HS diet affected the concentration of some milk FA, such as cis isomers of 18:1 and intermediates of ruminal biohydrogenation of polyunsaturated FA. Overall, bacterial DFM could be useful to stabilize ruminal pH. Their effects on CH4 production mitigation and milk FA profile depended on DFM strain and diet and should be confirmed under a greater variation of dietary conditions.

Additive methane-mitigating effect between linseed oil and nitrate fed to cattle.

The objective of this study was to test the effect of linseed oil and nitrate fed alone or in combination on methane (CH4) emissions and diet digestibility in cows. The experiment was conducted as a 2 × 2 factorial design using 4 multiparous nonlactating Holstein cows (initial BW 656 ± 31 kg). Each experimental period lasted 5 wk, with measures performed in the final 3 wk (wk 3 to 5). Diets given on a DM basis were 1) control (CON; 50% natural grassland hay and 50% concentrate), 2) CON with 4% linseed oil (LIN), 3) CON with 3% calcium nitrate (NIT), and 4) CON with 4% linseed oil plus 3% calcium nitrate (LIN+NIT). Diets were offered twice daily and were formulated to deliver similar amounts (DM basis) of CP (12.2%), starch (25.5%), and NDF (39.5%). Feed offer was restricted to 90% of voluntary intake (12.4 kg DMI/d). Total tract digestibility and N balance were determined from total feces and urine collected separately for 6 d during wk 4. Daily CH4 emissions were quantified using open chambers for 4 d during wk 5. Rumen fermentation and microbial parameters were analyzed from samples taken before and 3 h after the morning feeding. Rumen concentrations of dissolved hydrogen (H2) were measured continuously up to 6 h after feeding using a H2 sensor. Compared with the CON diet linseed oil and nitrate decreased (P < 0.01) CH4 emissions (g/kg DMI) by 17 and 22%, respectively, when fed alone and by 32% when combined. The LIN diet reduced CH4 production throughout the day, increased (P = 0.02) propionate proportion, and decreased (P = 0.03) ruminal protozoa concentration compared with CON diet. The NIT diet strongly reduced CH4 production 3 h after feeding, with a simultaneous increase in rumen dissolved H2 concentration, suggesting that nitrate does not act only as an electron acceptor. As a combined effect, linseed plus nitrate also increased H2 concentrations in the rumen. Diets had no effect (P > 0.05) on total tract digestibility of nutrients, except linseed oil, which tended to reduce (P < 0.10) fiber digestibility. Nitrogen balance (% of N intake) was positive for all diets but retention was less (P = 0.03) with linseed oil. This study demonstrates an additive effect between nitrate and linseed oil for reducing methanogenesis in cows without altering diet digestibility.

Amylase addition increases starch ruminal digestion in first-lactation cows fed high and low starch diets.

The objective of this study was to evaluate the effect of an exogenous amylase preparation on digestion of low- and high-starch diets in dairy cattle. Rumen and total-tract nutrient digestibility were measured in a 4×4 Latin square design with 28-d periods using 4 first-lactation cows cannulated at the rumen and duodenum. Corn silage-based diets had 20 or 30% starch, attained by changing the composition of concentrate, with or without addition of an exogenous amylase preparation. Effects of the enzyme additive were observed on ruminal digestibility but not at the total-tract level. Ruminal digestibility of starch increased from 75% in control to 81% with amylase supplementation. This difference in ruminal starch digestion was compensated postruminally, so that the total-tract digestibility of starch was almost complete and did not differ between treatments. The amylase supplement also increased the true ruminal digestibility of organic matter but did not affect microbial N flow to the duodenum. Amylase supplement reduced the proportion of acetate and butyrate and increased that of propionate, particularly in the high-starch diet, where it tended to increase the concentration of total volatile fatty acids in the rumen. Other effects were a higher amylase activity in the solid-associated microbial community and a tendency for lower numbers of protozoa. In contrast, we observed no changes in intake, production, dry matter and fiber (neutral detergent fiber and acid detergent fiber) digestibility, or ruminal digestion, and no or small changes on selected fibrolytic and amylolytic bacteria and on the microbial community in general. We conclude that the exogenous amylase improved starch digestion in the rumen in first-lactation cows with moderate intake and production levels.

Repeated acidosis challenges and live yeast supplementation shape rumen microbiota and fermentations and modulate inflammatory status in sheep.

This study aimed to investigate the impact of repeated acidosis challenges (ACs) and the effect of live yeast supplementation (Saccharomyces cerevisiae I-1077, SC) on rumen fermentation, microbial ecosystem and inflammatory response. The experimental design involved two groups (SC, n=6; Control, n=6) of rumen fistulated wethers that were successively exposed to three ACs of 5 days each, preceded and followed by resting periods (RPs) of 23 days. AC diets consisted of 60% wheat-based concentrate and 40% hay, whereas RPs diets consisted of 20% concentrate and 80% hay. ACs induced changes in rumen fermentative parameters (pH, lactate and volatile fatty-acid concentrations and proportions) as well as in microbiota composition and diversity. The first challenge drove the fermentation pattern towards propionate. During successive challenges, rumen pH measures worsened in the control group and the fermentation profile was characterised by a higher butyrate proportion and changes in the microbiota. The first AC induced a strong release of rumen histamine and lipopolysaccharide that triggered the increase of acute-phase proteins in the plasma. This inflammatory status was maintained during all AC repetitions. Our study suggests that the response of sheep to an acidosis diet is greatly influenced by the feeding history of individuals. In live yeast-supplemented animals, the first AC was as drastic as in control sheep. However, during subsequent challenges, yeast supplementation contributed to stabilise fermentative parameters, promoted protozoal numbers and decreased lactate producing bacteria. At the systemic level, yeast helped normalising the inflammatory status of the animals.

Behavioural adaptations of sheep to repeated acidosis challenges and effect of yeast supplementation.

This study aims to determine whether sheep modify their feeding and general behaviour when they undergo acidosis challenge, whether these modifications are maintained when acidosis challenges are repeated and whether yeast supplementation affects these modifications. Twelve rumen-cannulated wethers fed concentrate (wheat) and forage (hay) were exposed to three 28-day periods consisting of a 23-day recovery phase (20% of wheat) followed by a 5-day acidosis challenge (60% of wheat). Both diets limited food intake to 90% of ad libitum intake. Six sheep received a daily supplementation of a live yeast product, six received a placebo. Ruminal pH was recorded continuously. Daily consumption of wheat, hay, water and weekly consumption of salt were monitored. Behavioural observations were performed twice in each period: once under the recovery phase and once under acidosis challenge. These observations included video recordings over 24 h (time budget), social tests (mixing with another sheep for 5 min) and nociception tests (CO2 hot laser). As expected, sheep spent more time with a ruminal pH below 5.6 during challenges than during recovery phases (12.5 v. 4.7 h/day). Sheep drank more water (3.87 v. 3.27 l/day) and ingested more salt (16 v. 11 g/day) during challenges. They also spent more time standing than during recovery phases, adopting more frequent alarm postures and reacting more slowly to the hot stimulus. More severe behavioural modifications were observed during the first challenge than the two other challenges. Significant concentrate refusals were observed during challenge 1: from days 3 to 5 of this challenge, sheep ate only half of the distributed concentrate. Sheep were also more active and more aggressive towards each other in challenge 1. These behavioural modifications disappeared as the challenges were repeated: no behavioural modifications were observed between challenges and recovery phases during periods 2 and 3, and furthermore, sheep rapidly ate all the concentrate distributed during the third challenge. Focusing on the effects of yeast, the only differences registered between the two groups concerned ruminal pH, that is, mean ruminal pH values in the supplemented group were lower during the first challenge (5.11 v. 5.60) but higher during the third challenge (5.84 v. 5.28). In conclusion, our experiment suggests sheep can adapt to acidosis challenges, especially with yeast supplementation. Otherwise, ruminal pH values remained low during challenges, indicating that the modifications of general and feeding behaviour in subacute ruminal acidosis situations are not due exclusively to low ruminal pH values.

Experimental feed induction of ruminal lactic, propionic, or butyric acidosis in sheep.

A study was conducted to determine the feasibility to induce rumen acidosis with propionate, butyrate, or lactate as the major fermentation end products. Three rumen-cannulated Texel wethers were used in a 3 x 3 Latin square design. Each period consisted of 11 d of adaptation where wethers were daily fed at 90% of ad libitum intake a hay and wheat-based concentrate diet (4:1 ratio on a DM basis) in 2 equal portions followed by 3 d of acidosis induction. During the challenge, the morning feeding was replaced by an intraruminal supply of wheat (readily fermentable starch), corn (slowly fermentable starch), or beet pulp (easily digestible fiber), dosed at 1.2% of BW. Ruminal liquid samples were taken daily 1 h before (-1) and 1, 3, 5, and 6 h after intraruminal feed supply to measure pH, VFA, and lactic acid concentration. The differences between treatments accentuated throughout the 3-d challenge, being maximal and significant on d 3. Indeed, 6 h after the third day of the challenge, mean ruminal pH was less for wheat (4.85) than for corn (5.61; P = 0.008) and beet pulp (6.09; P = 0.001), and total VFA tended to be less for wheat (48.7 mM) than for corn and beet pulp (84.7 mM on average; P = 0.08). At the same time, the proportion of acetate was greater for wheat than for corn (75.5 and 62.2%, respectively; P = 0.005) but did not differ from beet pulp challenge (69.0%). The proportion of propionate was greatest for beet pulp compared with corn and wheat (21.0, 17.3, and 12.1%, respectively; P = 0.03), whereas the butyrate proportion was greatest for corn, intermediate for wheat, and least for beet pulp (16.3, 10.8, and 8.3%, respectively; P = 0.05). Lactate concentration was greatest for wheat (45.5 mM) compared with corn and beet pulp (8.3 mM on average; P = 0.01). Under our experimental conditions, ruminal lactic acidosis was successfully induced by wheat, whereas butyric and propionic subacute ruminal acidosis were respectively provoked by corn and beet pulp. We developed an original model that promoted differentiated fermentation pathways in the rumen of sheep. It will be used to study the ruminal microbiome changes involved in different acidosis situations.