A fibrolytic enzyme additive for lactating Holstein cow diets: ruminal fermentation, rumen microbial populations, and enteric methane emissions.

The objective was to determine if supplementing a dairy cow diet with an exogenous fibrolytic enzyme additive (Econase RDE; AB Vista, Marlborough, Wiltshire, UK) altered fermentation, pH, and microbial populations in the rumen or enteric methane (CH(4)) emissions. In a companion study, this enzyme additive improved efficiency of fat-corrected milk production in a dose-dependent manner by up to 11% for early lactation dairy cows. Nine ruminally cannulated, lactating Holstein cows were used in a replicated 3 × 3 Latin square design with 21-d periods. Dietary treatments were 0 (control), 0.5 (low), and 1.0 (high) mL of enzyme/kg of total mixed ration dry matter. Rumen contents were collected on 2 d (d 15 and 19), ruminal pH was measured continuously for 6 d (d 13 to 18) by using an indwelling system, and enteric CH(4) production was measured for 3 d (d 16 to 18) using the sulfur hexafluoride tracer gas technique. The enzyme additive did not alter volatile fatty acids, NH(3), pH, or population densities of total protozoa, bacteria, and methanogens in ruminal fluid. However, population densities of certain bacteria, calculated as copy number of species-specific 16S-rRNA, were affected by enzyme treatment. Population density of Ruminobacter amylophilus was increased and that of Fibrobacter succinogenes tended to be increased by the high enzyme treatment. Selenomonas ruminantium tended to increase linearly with increasing levels of enzyme in the diet, although its population density was only numerically increased by the high enzyme treatment. Streptococcus bovis, however, tended to be decreased by the low enzyme treatment. Increasing the level of enzyme supplement in the diet also linearly increased enteric CH(4) production, even when adjusted for feed intake or milk production (19.3, 20.8, and 21.7 g of CH(4)/kg of dry matter intake or 12.9, 13.6, and 15.1g of CH(4)/kg of milk for the control, low, and high enzyme treatments, respectively). This shift in ruminal bacterial communities and higher CH(4) emissions could imply increased ruminal digestion of feed, which needs to be substantiated in longer term studies.

Relationship between rumen methanogens and methane production in dairy cows fed diets supplemented with a feed enzyme additive.

AIMS: To investigate the relationship between ruminal methanogen community and host enteric methane (CH(4) ) production in lactating dairy cows fed diets supplemented with an exogenous fibrolytic enzyme additive. METHODS AND RESULTS: Ecology of ruminal methanogens from dairy cows fed with or without exogenous fibrolytic enzymes was examined using PCR-denaturing gradient gel electrophoresis (PCR-DGGE) analyses and quantitative real-time PCR (qRT-PCR). The density of methanogens was not affected by the enzyme additive or sampling times, and no relationship was observed between the total methanogen population and CH(4) yield (as g per head per day or g kg(-1) DMI). The PCR-DGGE profiles consisted of 26 distinctive bands, with two bands similar to Methanogenic archaeon CH1270 negatively correlated, and one band similar to Methanobrevibacter gottschalkii strain HO positively correlated, with CH(4) yield. Three bands similar to Methanogenic archaeon CH1270 or Methanobrevibacter smithii ATCC 35061 appeared after enzyme was added. CONCLUSIONS: Supplementing a dairy cow diet with an exogenous fibrolytic enzyme additive increased CH(4) yield and altered the composition of the rumen methanogen community, but not the overall density of methanogens. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study to identify the correlation between methanogen ecology and host CH(4) yield from lactating dairy cows.

Effect of parity and stage of lactation on feed sorting behavior of lactating dairy cows.

The objectives of this study were to determine if feed sorting differs between primiparous (PP) and multiparous (MP) cows, if sorting changes from the period of peak lactation to peak dry matter intake (DMI), and whether feed sorting affects efficiency of production. Data on DMI, milk production, feed sorting (particle size of offered and refused feed), and energy status (plasma nonesterified fatty acid, ß-hydroxybutyrate, insulin and glucose concentration) were collected on 30 PP and 30 MP lactating Holstein dairy cows, individually housed and fed in tie-stalls, during 3 wk (wk 2, 6, and 10) over 10 wk of a lactation study. Cows averaged 53, 81, and 109±10 d in milk (DIM) at the beginning of each of the 3 recording weeks. To determine sorting, feed samples were separated with a particle separator that had 3 screens (19, 8, and 1.18 mm) and a bottom pan, resulting in 4 fractions (long, medium, short, fine). Over the study period, MP cows consumed more DM and produced more milk than did the PP cows, but had similar efficiency of production (kg of milk/kg of DMI), and similar levels of plasma indicators of energy status. Across the study period, DMI increased, whereas milk yield decreased, resulting in decreased efficiency of milk production as cows moved further into lactation. All cows had higher nonesterified fatty acid and lower insulin concentrations in plasma at 53 DIM compared with at 81 and 109 DIM, suggesting they were mobilizing more body fat at that earlier stage of lactation. Across periods, all cows sorted against the longest ration particles, did not sort the medium ration particles, and sorted for fine ration particles; as a result, all cows consumed less neutral detergent fiber (NDF) and physically effective NDF (proportion of NDF retained on the 19- and 8-mm screens of the particle separator) than predicted. Greater selection against the longest ration particles was associated with greater efficiency of milk production; however, this sorting pattern also tended to be associated with lower milk fat percentage. The extent of the observed sorting against the longest, physically effective ration particles and for the finest ration particles was greater for PP cows than for MP cows across all 3 observation periods. Feed sorting remained consistent in cows across the DIM evaluated in this study and that this sorting behavior can affect the efficiency of milk production as well as milk fat percentage. Further, PP cows engage in more sorting of their ration than MP cows.

Improved milk production efficiency in early lactation dairy cattle with dietary addition of a developmental fibrolytic enzyme additive.

A 3-part study was conducted to evaluate the effect of a developmental fibrolytic enzyme additive on the digestibility of selected forages and the production performance of early-lactation dairy cows. In part 1, 4 replicate 24-h batch culture in vitro incubations were conducted with alfalfa hay, alfalfa silage, and barley silage as substrates and ruminal fluid as the inoculum. A developmental fibrolytic enzyme additive (AB Vista, Marlborough, UK) was added at 5 doses: 0, 0.5, 1.0, 1.5, and 2.0 µL/g of forage dry matter (DM). After the 24-h incubation, DM, neutral detergent fiber (NDF), and acid detergent fiber (ADF) disappearance were determined. For alfalfa hay, DM, NDF, and ADF disappearance was greater at the highest dosage compared with no enzyme addition. Barley silage NDF and ADF and alfalfa silage NDF disappearance tended to be greater for the highest enzyme dosage compared with no enzyme addition. In part 2, 6 ruminally cannulated, lactating Holstein dairy cows were used to determine in situ degradation of alfalfa and barley silage, with (1.0 mL/kg of silage DM) and without added enzyme. Three cows received a control diet (no enzyme added) and the other 3 received an enzyme-supplemented (1.0 mL/kg of diet DM) diet. Enzyme addition after the 24h in situ incubation did not affect the disappearance of barley silage or alfalfa silage. In part 3, 60 early-lactation Holstein dairy cows were fed 1 of 3 diets for a 10-wk period: (1) control (CTL; no enzyme), (2) low enzyme (CTL treated with 0.5 mL of enzyme/kg of diet DM), and (3) high enzyme (CTL treated with 1.0 mL of enzyme/kg of diet DM). Adding enzyme to the diet had no effect on milk yield, but dry matter intake was lower for the high enzyme treatment and tended to be lower for the low enzyme treatment compared with CTL. Consequently, milk production efficiency (kg of 3.5% fat-corrected milk/kg of DM intake) linearly increased with increasing enzyme addition. Cows fed the low and high enzyme diets were 5.3 (not statistically significant) and 11.3% more efficient, respectively, compared with CTL cows. This developmental fibrolytic enzyme additive has the potential to increase fiber digestibility of forages, which could lead to greater milk production efficiency for dairy cows in early lactation.

Crushed sunflower, flax, or canola seeds in lactating dairy cow diets: effects on methane production, rumen fermentation, and milk production.

The objective of this study was to investigate the potential of reducing enteric methane production from dairy cows by incorporating into the diet various sources of long-chain FA varying in their degree of saturation and ruminal availability. The experiment was conducted as a crossover design with 16 lactating dairy cows maintained in 2 groups and fed 4 dietary treatments in four 28-d periods. Eight ruminally cannulated primiparous cows (96 +/- 18 d in milk) were assigned to group 1 and 8 multiparous cows (130 +/- 31 d in milk) were assigned to group 2. The dietary treatments were: 1) a commercial source of calcium salts of long-chain fatty acids (CTL), 2) crushed sunflower seeds (SS), 3) crushed flaxseed (FS), and 4) crushed canola seed (CS). The oilseeds added 3.1 to 4.2% fat to the diet (DM basis). All 3 oilseed treatments decreased methane production (g/d) by an average of 13%. When corrected for differences in dry matter intake (DMI), compared with CTL, methane production (g/kg of DM intake) was decreased by feeding FS (-18%) or CS (-16%) and was only numerically decreased (-10%) by feeding SS. However, compared with the CTL, feeding SS or FS lowered digestible DMI by 16 and 9%, respectively, because of lowered digestibility. Thus, only CS lowered methane per unit of digestible DM intake. Feeding SS and CS decreased rumen protozoal counts, but there were no treatment effects on mean ruminal pH or total volatile fatty acid concentration. Milk efficiency (3.5% fat corrected milk/DMI), milk yield, and component yield and concentrations were not affected by oilseed treatments. The study shows that adding sources of long-chain fatty acids to the diet in the form of processed oilseeds can be an effective means of reducing methane emissions. However, for some oilseeds such as SS or FS, the reduction in methane can be at the expense of diet digestibility. The use of crushed CS offers a means of mitigating methane without negatively affecting diet digestibility, and hence, milk production.

Feeding saponin-containing Yucca schidigera and Quillaja saponaria to decrease enteric methane production in dairy cows.

An experiment was conducted in vitro to determine whether the addition of saponin-containing Yucca schidigera or Quillaja saponaria reduces methane production without impairing ruminal fermentation or fiber digestion. A slightly lower dose of saponin was then fed to lactating dairy cows to evaluate effects on ruminal fermentation, methane production, total-tract nutrient digestibility, and milk production and composition. A 24-h batch culture in vitro incubation was conducted in a completely randomized design with a control (no additive, CON) and 3 doses of either saponin source [15, 30, and 45 g/kg of substrate dry matter (DM)] using buffered ruminal fluid from 3 dairy cows. The in vivo study was conducted as a crossover design with 2 groups of cows, 3 treatments, and three 28-d periods. Six ruminally cannulated cows were used in group 1 and 6 intact cows in group 2 (627 +/- 55 kg of body weight and 155 +/- 28 d in milk). The treatments were 1) early lactation total mixed ration, no additive (control; CON); 2) CON diet supplemented with whole-plant Y. schidigera powder at 10 g/kg of DM (YS); and 3) CON diet supplemented with whole-plant Q. saponaria powder at 10 g/kg of DM (QS). Methane production was measured in environmental chambers and with the sulfur hexafluoride (SF(6)) tracer technique. In vitro, increasing levels of both saponin sources decreased methane concentration in the headspace and increased the proportion of propionate in the buffered rumen fluid. Concentration of ammonia-N, acetate proportion, and the acetate:propionate ratio in the buffered rumen fluid as well as 24-h digestible neutral detergent fiber were reduced compared with the CON treatment. Medium and high saponin levels decreased DM digestibility compared with the CON treatment. A lower feeding rate of both saponin sources (10 g/kg of DM) was used in vivo in an attempt to avoid potentially negative effects of higher saponin levels on feed digestibility. Feeding saponin did not affect milk production, total-tract nutrient digestibility, rumen fermentation, or methane production. However, DM intake was greater for cows fed YS and QS than for CON cows, with a tendency for greater DM intake for cows fed YS compared with those fed QS. Consequently, efficiency of milk production (kg of milk/kg of DM intake) was lower for cows fed saponin compared with controls. The results show that although saponin from Y. schidigera and Q. saponaria lowered methane production in vitro, the reduction was largely due to reduced ruminal fermentation and feed digestion. Feeding a lower dose of saponin to lactating dairy cows avoided potentially negative effects on ruminal fermentation and feed digestion, but methane production was not reduced. Lower efficiency of milk production of cows fed saponin, and potential reductions in feed digestion at high supplementation rates may make saponin supplements an unattractive option for lowering methane production in vivo.

Feeding behavior and ruminal pH of corn silage, barley grain, and corn dried distillers' grain offered in a total mixed ration or in a free-choice diet to beef cattle.

Seventy-nine continental crossbred beef heifers (524.4 ± 41.68 kg BW), 16 of which were ruminally cannulated, were used in a 53-d experiment with a generalized randomized block design to assess the effects of barley grain (BG), corn silage (CS), and corn distillers' grain (DG) offered in a free-choice diet on feeding behavior and ruminal fermentation. Treatments were total mixed ration (TMR) consisting of 85% BG, 10% CS, and 5% supplement or free-choice (i.e., self-selection) diets of BG and CS (BGCS), BG and corn dry DG (BGDG), or CS and corn DG (CSDG). Heifers were housed in groups of 9 or 10 in 8 pens and weighed 2 h before feed delivery at d 0, 21, 42, and 52 of the study. Pens were equipped with an electronic feed bunk monitoring system enabling feed intake and feeding behavior to be continuously monitored. Each of these pens was randomly allocated 2 cannulated heifers equipped with indwelling pH probes for continuous measurement of ruminal pH during wk 1, 2, 4, and 7. Blood and rumen contents were taken from cannulated heifers 2 h after feed delivery on d -3, 0, 7, 8, 42, and 49. Cattle fed either TMR or free-choice diets had similar (P > 0.10) ruminal fermentation, blood profile, and growth performance, with the exception of the CSDG diet, for which ruminal pH levels were consistently greater (P < 0.01) and performance was lower (P < 0.01). When DG was a component in free-choice diets, heifers reduced its inclusion in the diet (P < 0.05) over the experiment without affecting growth rate or ruminal fluid pH. Finishing feedlot cattle fed BG and CS separately selected a diet with a greater proportion of BG (85% DMI) compared to the TMR with no signs of acidosis. When cattle were given free-choice access to corn dry DG as an alternative to CS, they consumed levels up to 30% of their total daily DMI. Under the conditions of our experiment cattle can effectively self-select diets without increasing the risk of subclinical acidosis and still maintain similar levels of growth and feed efficiency compared with a TMR.

Feeding behavior and ruminal acidosis in beef cattle offered a total mixed ration or dietary components separately.

Eighty continental crossbred beef heifers (414.9 ± 37.9 kg of BW), 16 of which were ruminally cannulated, were used in a 52-d experiment with a generalized randomized block design, to assess if self-selection of dietary ingredients modulates ruminal pH and improves rumen function of feedlot finishing cattle. Treatments were total mixed ration [TMR; 85% barley grain (BG), 10% corn silage (CS), 5% supplement]; or free-choice (self-selection; FC) diets of barley grain and corn silage (BGCS), barley grain and wheat distillers grain (BGDG), or corn silage and wheat distillers grain (CSDG). Heifers were housed in groups of 10 in 8 pens equipped with the GrowSafe System (Airdrie, AB, Canada) enabling feed intake and feeding behavior to be continuously monitored. Two cannulated heifers were randomly assigned to each pen and equipped with indwelling pH probes for continuous measurement of ruminal pH during 4 periods (d 1 to 4, d 7 to 14, d 21 to 28, and d 42 to 49). Rumen fluid samples were collected from cannulated heifers on d 7 and 42 before feed delivery, and on d 4 and 49 at 2 h post-feed delivery for determination of VFA. Heifers fed the TMR had shorter (P = 0.01) and smaller (P = 0.03) meals than those fed the FC diets. Cattle fed BGCS and BGDG increased (P < 0.01) intake of BG over time by up to 80 and 70%, respectively. Increased consumption of BG arose from an increase (P < 0.01) in eating rate over the same (P > 0.10) feeding time, which was accompanied by an increase (P < 0.05) in eating rate but a decrease (P < 0.05) in feeding time of either CS or DG. Even with increased BG consumption, ruminal pH and VFA profiles were not different (P > 0.10) among FC diets or compared with the TMR. Cattle fed FC CSDG consumed DG at 60% of dietary DM over the trial, resulting in greater (P < 0.05) mean ruminal pH and acetate-to-propionate ratio and less (P < 0.05) area under the curve than those given the other FC diets or the TMR. Finishing feedlot cattle fed FC diets containing BG self-regulate intake of diets that have a similar composition, intake level, and ruminal fermentation profile to those fed a TMR.