Dietary supplementation with rumen-protected capsicum during the transition period improves the metabolic status of dairy cows.

In ruminants, it has been observed that capsicum oleoresin can alter insulin responses and that high-intensity artificial sweetener can increase glucose absorption from the small intestine. Because glucose metabolism and insulin responses are critical during early lactation, these supplements might have an effect on the metabolic status of dairy cows during the transition period. The objective of this experiment was to evaluate the effects of rumen-protected capsicum oleoresin fed alone or in combination with artificial sweetener during the transition period on lactational performance and susceptibility to subclinical ketosis in dairy cows. Fifteen primiparous and 30 multiparous Holstein cows (a total of 39 cows finished the study) were arranged in a randomized complete block design during d -21 to 60 relative to parturition. Cows within block were randomly assigned to one of the following treatments: no supplement (CON), supplementation with 100 mg of rumen-protected capsicum/cow per day (RPCap), or RPCap plus 2 g of high-intensity artificial sweetener/cow per day (RPCapS). For both the RPCap and RPCapS treatments, only rumen-protected capsicum was fed during the dry period. From d 8 to 11 of lactation, intake was limited to 70% of predicted dry matter intake to induce subclinical ketosis. Production variables were recorded daily, samples for milk composition were collected on wk 2, 4, 6, and 8, and blood samples were collected on wk -2, 1, 2, and 4 of the experiment for analysis of metabolic hormones and blood cell counts. Supplementation with rumen-protected capsicum increased serum insulin and decreased ß-hydroxybutyrate concentrations precalving, indicating a decrease in lipolysis. During the lactation period, RPCap was associated with a trend for increased milk production and feed efficiency following the ketosis challenge. Supplementation with RPCapS appeared to negate the response to rumen-protected capsicum. All cows developed subclinical ketosis during the challenge, and this was not affected by treatment. We conclude that treatments did not decrease susceptibility to subclinical ketosis; however, dietary supplementation with rumen-protected capsicum was effective at improving energy status precalving and tended to increased milk production and feed efficiency. The mechanism underlying these responses is unclear.

Short communication: Short-term effect of 3-nitrooxypropanol on feed dry matter intake in lactating dairy cows.

The objective of this study was to investigate the effect of 3-nitrooxypropanol (3-NOP), an enteric methane inhibitor under investigation, on short-term dry matter intake (DMI) in lactating dairy cows. Following a 1-wk adaptation period, 12 multiparous Holstein cows were fed a basal total mixed ration (TMR) containing increasing levels of 3-NOP during 5 consecutive, 6-d periods. The experiment was conducted in a tiestall barn. Feed bins were split in half by a solid divider, and cows simultaneously received the basal TMR supplemented with the following: (1) a placebo without 3-NOP or (2) 3-NOP included in the TMR at 30, 60, 90, or 120 mg/kg of feed dry matter (experimental periods 2, 3, 4, and 5, respectively). Cows received the control diet (basal TMR plus placebo premix) during experimental period 1. A premix containing ground corn grain, soybean oil, and dry molasses was used to incorporate 3-NOP in the ration. Cows were fed twice daily as follows: 60% of the daily feed allowance at 0800 h and 40% at 1800 h. Feed offered and refused was recorded at each feeding. During the morning feedings, each cow was offered either control or 3-NOP-treated TMR at 150% of her average intake during the previous 3 d. After collection of the evening refusals, cows received only the basal TMR without the premix until the next morning feeding. The test period for the short-term DMI data collection was defined from morning feeding to afternoon refusals collection during each day of each experimental period. Location (left or right) of the control and 3-NOP diets within a feed bin was switched every day during each period to avoid feed location bias. Dry matter intake of TMR during the test period was quadratically increased by 3-NOP compared with the control. Inclusion of 3-NOP at 120 mg/kg of feed dry matter resulted in decreased 10-h DMI compared with the lower 3-NOP doses, but was similar to the control. There was no effect of feed location (left or right) within feed bin on DMI. Data from this short-term study suggests that 3-NOP does not have a negative effect on DMI in lactating dairy cows.

Dose-response effect of 3-nitrooxypropanol on enteric methane emissions in dairy cows.

This experiment was designed to test the effect of inclusion rate of 3-nitrooxypropanol (3-NOP), a methane inhibitor, on enteric methane emissions in dairy cows. The study was conducted with 49 multiparous Holstein cows in a randomized complete block design in 2 phases; phase 1 was with 28 cows, and phase 2 with 21 cows. Cows were fed a basal total mixed ration ad libitum and were blocked based on days in milk, milk yield, and enteric methane emissions during a 14-d covariate period. Treatments were control (no 3-NOP) and 40, 60, 80, 100, 150, and 200 mg of 3-NOP/kg of feed dry matter. Following a 14-d adaptation period, enteric gaseous emissions (methane, carbon dioxide, and hydrogen) were measured using the GreenFeed system (C-Lock Inc., Rapid City, SD) over a 3-d period. Compared with the control, inclusion rate of 3-NOP quadratically decreased daily enteric methane emissions from 22 to 40%. Maximum mitigation effect was achieved with the 3 highest 3-NOP doses (with no statistical difference among 100, 150, and 200 mg/kg). The decrease in methane emission yield and emission intensity ranged from 16 to 36% and from 25 to 45%, respectively. Emissions of hydrogen quadratically increased 6- to 10-fold, compared with the control; the maximum increase was with 150 mg/kg 3-NOP. Treatment did not affect daily emissions of carbon dioxide, but a linear increase in carbon dioxide emission yield was observed with increasing 3-NOP doses. Dry matter intake and milk yield of the cows was not affected by 3-NOP. Milk fat concentration and yield were increased by 3-NOP due to increased concentration of de novo synthetized short-chain fatty acids in milk. Inclusion of 3-NOP also tended to increase milk urea nitrogen but had no other effects on milk components. In this short-term experiment, 3-NOP decreased enteric methane emissions without affecting dry matter intake or milk yield and increased milk fat in dairy cows. Maximum mitigation effect was achieved at 100 to 200 mg/kg of feed dry matter.

Effects of 3-nitrooxypropanol on rumen fermentation, lactational performance, and resumption of ovarian cyclicity in dairy cows.

This study examined the effect of 3-nitrooxypropanol (3-NOP), a substance under investigation, on enteric methane (CH4) emission, rumen fermentation, lactational performance, sensory properties of milk, and the resumption of ovarian cyclicity in early-lactation dairy cows. Fifty-six multi- and primiparous Holstein cows, including 8 that were rumen cannulated, were used in a 15-wk randomized complete block design experiment. Cows were blocked based on parity and previous lactation milk yield (MY) or predicted MY, and within each block were randomly assigned to one of 2 treatments: (1) control (CON), administered no 3-NOP, or (2) 3-NOP applied at 60 mg/kg of feed dry matter (3-NOP). Enteric CH4 emission was measured during experimental wk 2, 6, 9, and 15, using the GreenFeed system. Dry matter intake (DMI) and MY data were collected daily throughout the experiment, and milk composition samples were collected 7 times during the experiment. Milk samples were collected from 14 to 60 (±2) d after calving, 3 d per week, and assayed for progesterone concentration to determine resumption of ovarian activity. Compared with CON, 3-NOP decreased daily CH4 emission by 26%, CH4 yield (CH4 per kg of DMI) by 21%, and CH4 emission intensity [CH4 per kg of MY or energy-corrected milk (ECM)] by 25%. Enteric emission of carbon dioxide was decreased by 5%, and hydrogen emission was increased 48-fold by 3-NOP. Inclusion of 3-NOP decreased concentration of total volatile fatty acids (by 9.3%) and acetate but increased butyrate molar proportion, ethanol, and formate concentrations in ruminal fluid. Dry matter intake was lower for 3-NOP compared with CON, but DMI expressed as a percentage of body weight was not different between treatments. Treatment had no effect on milk and ECM, body weight change, or body condition score. Milk composition and milk fat and protein yields were not affected by treatment, except that concentrations of short-chain fatty acids in milk were increased by 3-NOP. Nutrient digestibility and blood metabolites and hormones were not affected by 3-NOP, except that insulin was decreased by 3-NOP. There was no effect of 3-NOP on postpartum resumption of ovarian activity, including days to first and second luteal phases, length of first and second luteal phases, and interval from first to second luteal phase. Sensory properties of milk from cows fed 3-NOP and cheese made from that milk were not affected by treatment. In this experiment, 3-NOP decreased daily enteric CH4 emission, emission yield, and emission intensity, improved feed efficiency, and did not affect lactational performance or onset of ovarian activity in early-lactation dairy cows.

Effects of ensiling time on corn silage neutral detergent fiber degradability and relationship between laboratory fiber analyses and in vivo digestibility.

Accurate analysis of degradability of silage neutral detergent fiber (NDF) is important for diet formulation and to predict lactational performance of dairy cows. In this study, 5 corn silage hybrids ensiled for 0 (unfermented), 30, 60, 120, and 150 d were used to determine the effects of ensiling time on silage neutral detergent fiber degradability (NDFD) and to assess the relationships between near-infrared reflectance spectroscopy (NIR) NDF-related analyses and in situ NDFD variables. In addition, the relationships between dietary concentration of indigestible NDF, 288-h incubation (iNDF288), or undegraded NDF, 240-h incubation (uNDF240), and in vivo total-tract apparent organic matter and NDF digestibility were studied in total mixed ration samples from 16 experiments with lactating dairy cows. Ensiling time had no effect on silage NDF concentration; however, the ratio of acid detergent fiber ÷ NDF increased, and estimated hemicellulose concentration decreased quadratically with ensiling time. Also, concentration of NDF-bound protein decreased, and that of lignin increased linearly with ensiling time. These changes in silage fiber composition resulted in a linear decrease in in situ effective degradability of silage NDF with increasing ensiling time. The indigestible fraction of NDF and concentration of structural carbohydrates were not affected by ensiling time. Correlations of in situ NDFD variables with laboratory NIR NDFD analyses were weak to moderate. The relationship of corn silage uNDF240 with lignin concentration or 30-h NDFD (all NIR analyses) was remarkably good (R2 = 0.73 and 0.88, respectively). The relationship between in situ iNDF288 concentration (but not uNDF240) and in vivo total-tract apparent digestibility of dietary organic matter and NDF was good (R2 = 0.72 and 0.80, respectively). In conclusion, in situ degradability of silage NDF linearly decreased from 0 to 150 d ensiling time, primarily caused by a decrease in concentrations of hemicellulose and NDF-bound protein. In situ NDF degradability measurements and common laboratory NIR NDF-related analyses were generally poorly correlated. We found a good relationship between in vivo NDF digestibility and dietary concentration of iNDF288 determined in situ, but the relationship with uNDF240 was poor.

Production effects of feeding extruded soybean meal to early-lactation dairy cows.

The objective of this experiment was to evaluate productive and reproductive effects of replacing solvent-extracted soybean meal (SSBM) with extruded soybean meal (ESBM) in a total mixed ration for early-lactation dairy cows. Thirty-four Holstein cows (12 primiparous and 22 multiparous) were used in a randomized complete block design experiment with 17 cows per treatment. Feeding was ad libitum for 5 to 10% refusals. A fresh-cow diet was fed the first 21 d in milk followed by a lactation diet from 22 to 60 d in milk. Milk and dry matter intake data were collected throughout the experiment, and samples were collected for blood chemistry and amino acid profile, nutrient digestibility, nitrogen utilization, and enteric methane emission using the GreenFeed system (C-Lock Inc., Rapid City, SD). Dry matter intake, milk yield, and feed efficiency were not different between SSBM and ESBM. Energy-corrected milk yield and efficiency were also not different between diets. Diet had no effect on milk composition, except that milk true protein yield was decreased by ESBM. Enteric methane emission, yield, and intensity were not different between SSBM and ESBM. Because of its greater fat content, ESBM triggered expected changes in milk fatty acid (FA) profile: decreased sum of C16, saturated, and odd- and branched-chain FA and increased sum of preformed FA, polyunsaturated, and trans FA. The ESBM diet increased or tended to increase some essential amino acids in plasma. In this study, ESBM did not affect dry matter intake and did not improve lactational performance or onset of ovarian function in early-lactation dairy cows, and it decreased milk protein yield, possibly due to greater unsaturated FA intake compared with SSBM.

Acceptance of flavored concentrate premixes by young ruminants following a short-term exposure.

Three cafeteria feeding design experiments were conducted to test whether young ruminants have flavor preferences. Experiment 1 was with 11 Dorset × Suffolk weaned lambs of both sexes, aged 5 mo and averaging 47.5 (standard deviation = 5.8) kg of body weight. The lambs were offered a choice of 5 flavored concentrate premixes (FCP) and an unflavored control for 5 min 4 times over 10 d. The FCP were prepared by mixing 200 to 300 g/t (as-is basis) of synthetic flavors (vanilla, milky, spicy/fenugreek, red summer fruits, and molasses) into a basal diet. The unflavored control and the milky flavor were consumed in greater amounts than all other flavors at 83.9 and 65.8 g/test, respectively. The consumption rate of FCP (g/min) was similar among treatments. Lambs spent more time consuming the milky flavor and the control at 123 and 144 s/test, respectively, compared with all other FCP (average of 65 s/test). In experiment 2, 12 weaned female Holstein Friesian calves (56-68 d of age) averaging 75.8 kg (standard deviation = 8.45) of body weight were offered a choice of 4 FCP (vanilla, milky, spicy/fenugreek, and red summer fruits) at an inclusion rate of 150 to 200 g/t (as-is basis) and the unflavored control for 5 min 4 times over 10 d. The average consumption rate was 27.8 g/min, and there were no differences among FCP. In experiment 3, a choice of 4 FCP with 2 different flavor combinations (vanilla-fenugreek and milky-vanilla) included at 75 g/t (as-is basis; low) or 150 g/t (high) was offered to a total of 12 weaned female Holstein Friesian calves (47-62 d of age) with an average body weight of 65.3 kg (standard deviation = 7.91). The FCP were offered daily for 14 d for 30 to 60 min/d. Vanilla-fenugreek (low) was consumed less at 57.5 g/test per calf compared with the other FCP (average of 87.5 g/test per calf). There were no other differences among FCP in experiment 3. Overall, compared with the control, flavors used in the present experiments did not affect feed intake of weaned lambs and calves. Other factors, such as taste, sight, texture of the feed, effect of the dams as previous experience (via maternal ingestion, which influences neonatal feeding), and their interactions, may also play a role in flavor preferences of young ruminants.

Inclusion of brown midrib dwarf pearl millet silage in the diet of lactating dairy cows.

Brown midrib brachytic dwarf pearl millet (Pennisetum glaucum) forage harvested at the flag leaf visible stage and subsequently ensiled was investigated as a partial replacement of corn silage in the diet of high-producing dairy cows. Seventeen lactating Holstein cows were fed 2 diets in a crossover design experiment with 2 periods of 28 d each. Both diets had forage:concentrate ratios of 60:40. The control diet (CSD) was based on corn silage and alfalfa haylage, and in the treatment diet, 20% of the corn silage dry matter (corresponding to 10% of the dietary dry matter) was replaced with pearl millet silage (PMD). The effects of partial substitution of corn silage with pearl millet silage on dry matter intake, milk yield, milk components, fatty acid profile, apparent total-tract digestibility of nutrients, N utilization, and enteric methane emissions were analyzed. The pearl millet silage was higher in crude protein and neutral detergent fiber and lower in lignin and starch than the corn silage. Diet did not affect dry matter intake or energy-corrected milk yield, which averaged 46.7 ± 1.92 kg/d. The PMD treatment tended to increase milk fat concentration, had no effect on milk fat yield, and increased milk urea N. Concentrations and yields of milk protein and lactose were not affected by diet. Apparent total-tract digestibility of dry matter decreased from 66.5% in CSD to 64.5% in PMD. Similarly, organic matter and crude protein digestibility was decreased by PMD, whereas neutral- and acid-detergent fiber digestibility was increased. Total milk trans fatty acid concentration was decreased by PMD, with a particular decrease in trans-10 18:1. Urinary urea and fecal N excretion increased with PMD compared with CSD. Milk N efficiency decreased with PMD. Carbon dioxide emission was not different between the diets, but PMD increased enteric methane emission from 396 to 454 g/d and increased methane yield and intensity. Substituting corn silage with brown midrib dwarf pearl millet silage at 10% of the diet dry matter supported high milk production in dairy cows. When planning on farm forage production strategies, brown midrib dwarf pearl millet should be considered as a viable fiber source.

Inclusion of wheat and triticale silage in the diet of lactating dairy cows.

The objective of this experiment was to partially replace corn silage with 2 alternative forages, wheat (Triticum aestivum) or triticale (X Triticosecale) silages at 10% of the diet dry matter (DM), and investigate the effects on dairy cow productivity, nutrient utilization, enteric CH4 emissions, and farm income over feed costs. Wheat and triticale were planted in the fall as cover crops and harvested in the spring at the boot stage. Neutral- and acid-detergent fiber and lignin concentrations were higher in the wheat and triticale silages compared with corn silage. The forages had similar ruminal in situ effective degradability of DM. Both alternative forages had 1% starch or less compared with the approximately 35% starch in corn silage. Diets with the alternative forages were fed in a replicated 3 × 3 Latin square design experiment with three 28-d periods and 12 Holstein cows. The control diet contained 44% (DM basis) corn silage. In the other 2 diets, wheat or triticale silages were included at 10% of dietary DM, replacing corn silage. Dry matter intake was not affected by diet, but both wheat and triticale silage decreased yield of milk (41.4 and 41.2 vs. 42.7 ± 5.18 kg/d) and milk components, compared with corn silage. Milk fat from cows fed the alternative forage diets contained higher concentrations of 4:0, 6:0, and 18:0 and tended to have lower concentrations of total trans fatty acids. Apparent total-tract digestibility of DM and organic matter was decreased in the wheat silage diet, and digestibility of neutral-and acid-detergent fiber was increased in the triticale silage diet. The wheat and triticale silage diets resulted in higher excretion of urinary urea, higher milk urea N, and lower milk N efficiency compared with the corn silage diet. Enteric CH4 emission per kilogram of energy-corrected milk was highest in the triticale silage diet, whereas CO2 emission was decreased by both wheat and triticale silage. This study showed that, at milk production of around 42 kg/d, wheat silage and triticale silage can partially replace corn silage DM and not affect DM intake, but milk yield may decrease slightly. For dairy farms in need of more forage, triticale or wheat double cropped with corn silage may be an appropriate cropping strategy.

Using brown midrib 6 dwarf forage sorghum silage and fall-grown oat silage in lactating dairy cow rations.

Double cropping and increasing crop diversity could improve dairy farm economic and environmental sustainability. In this experiment, corn silage was partially replaced with 2 alternative forages, brown midrib-6 brachytic dwarf forage sorghum (Sorghum bicolor) or fall-grown oat (Avena sativa) silage, in the diet of lactating dairy cows. We investigated the effect on dry matter (DM) intake, milk yield (MY), milk components and fatty acid profile, apparent total-tract nutrient digestibility, N utilization, enteric methane emissions, and income over feed cost. We analyzed the in situ DM and neutral detergent fiber disappearance of the alternative forages versus corn silage and alfalfa haylage. Sorghum was grown in the summer and harvested in the milk stage. Oats were grown in the fall and harvested in the boot stage. Compared with corn silage, neutral detergent fiber and acid detergent fiber concentrations were higher in the alternative forages. Lignin content was highest for sorghum silage and similar for corn silage and oat silage. The alternative forages had less than 1% starch compared with the approximately 35% starch in the corn silage. Ruminal in situ DM effective degradability was similar, although statistically different, for corn silage and oat silage, but lower for sorghum silage. Diets with the alternative forages were fed in a replicated 3 × 3 Latin square design experiment with three 28-d periods and 12 Holstein cows. The control diet contained 44% (DM basis) corn silage. In the other 2 diets, sorghum or oat silages were included at 10% of dietary DM, replacing corn silage. Sorghum silage inclusion decreased DM intake, MY, and milk protein content but increased milk fat and maintained energy-corrected MY similar to the control. Oat silage had no effect on DM intake, MY, or milk components compared to the control. The oat silage diet increased apparent total-tract digestibility of dietary nutrients, except starch, whereas the sorghum diet slightly decreased DM, organic matter, crude protein, and starch digestibility. Cows consuming the oat silage diet had higher milk urea N and urinary urea N concentrations. Milk N efficiency was decreased by the sorghum diet. Diet did not affect enteric methane or carbon dioxide emissions. This study shows that oat silage can partially replace corn silage at 10% of the diet DM with no effect on MY. Brown midrib sorghum silage harvested at the milk stage with <1% starch may decrease DM intake and MY in dairy cows.

Histidine deficiency has a negative effect on lactational performance of dairy cows.

A 10-wk randomized complete block design experiment with 24 Holstein cows was conducted to investigate the long-term effects of feeding a His-deficient diet on lactational performance of dairy cows. Cows were blocked by days in milk, milk yield, and parity, and randomly assigned to 1 of the following 2 treatments: (1) His-adequate diet [HAD; providing +166 g/d over metabolizable protein (MP) requirements, according to the National Research Council (2001) and digestible His (dHis) supply of 68 g/d, or 2.5% of MP requirements] and (2) His-deficient diet (HDD; +37 g/d over MP requirements and dHis supply of 49 g/d, or 1.9% of MP requirements). Both HAD and HDD were supplemented with rumen-protected (RP) Met and Lys supplying digestible Met and digestible Lys at 2.4 and 2.4% and 7.2 and 7.1% of MP requirements, respectively. At the end of the 10-wk experiment, HDD was supplemented with RPHis (HDD+RPHis; total dHis supply of 61 g/d, or 2.4% of MP requirements) for an additional 9 d. Dry matter intake (DMI; 25.4 and 27.1 kg/d, standard error of the mean = 0.41), yields of milk (37.6 and 40.5 kg/d, standard error of the mean = 0.62), protein and lactose, energy-corrected milk, and milk and plasma urea-N were decreased by HDD compared with HAD. Feed and energy-corrected milk feed efficiencies, milk fat, protein and lactose concentrations, body weight, and body condition score of the cows were not affected by treatment. Apparent total-tract digestibility of dry and organic matter, crude protein, and neutral detergent fiber, and excretion of urinary N and urea-N were decreased by HDD compared with HAD. Concentration of plasma leptin tended to be decreased for HDD compared with HAD. Plasma concentrations of EAA (His, Leu, Lys, Val) and carnosine decreased and total EAA tended to be decreased in cows fed HDD compared with HAD. Muscle concentrations of free His, Leu, and Val decreased and Gly and ß-alanine tended to be increased by HDD compared with HAD. Cows fed HDD had a lower blood hemoglobin concentration than cows fed HAD. At the end of the 10-wk study, the 9-d supplementation of HDD with RPHis (i.e., HDD+RPHis) increased DMI and plasma His, and tended to increase energy-corrected milk yield and plasma carnosine, compared with HDD. In conclusion, feeding a diet deficient in dHis supplying adequate MP, digestible Met, and digestible Lys affected negatively lactational performance of dairy cows. These results confirm our previous findings that low dietary His supply can impair DMI, yields of milk and milk protein, and blood hemoglobin in dairy cows.

Effect of high-oleic-acid soybeans on production performance, milk fatty acid composition, and enteric methane emission in dairy cows.

The objective of this study was to investigate the effect of 3 soybean sources differing in fatty acid profile and processing method on productivity, milk composition, digestibility, rumen fermentation, and enteric methane emission in lactating dairy cows. The soybean sources were conventional, high-linoleic-acid variety extruded soybean meal (ESBM; 8.7% ether extract with 15% oleic and 54% linoleic acids); extruded Plenish (DuPont Pioneer, Johnston, IA), high-oleic-acid variety soybean meal (EPSBM; 8.4% ether extract with 73% oleic and 8% linoleic acids); and whole, heated Plenish soybeans (WPSB; 20.2% ether extract). The study involved 15 Holstein cows in a replicated 3 × 3 Latin square design experiment with three 28-d periods. The inclusion rate of the soybean sources in the diet was (dry matter basis) 17.1, 17.1, and 7.4% for ESBM, EPSBM, and WPSB, respectively, which resulted in ether extract concentration of the diets of 3.99, 3.94, and 4.18%, respectively. Compared with ESBM, the Plenish diets tended to increase dry matter intake and decreased feed efficiency (but had no effect on energy-corrected milk feed efficiency). The Plenish diets increased milk fat concentration on average by 5.6% and tended to increase milk fat yield, compared with ESBM. The WPSB diet tended to increased milk true protein compared with the extruded soybean meal diets. Treatments had no effect on rumen fermentation and enteric methane or carbon dioxide emissions, except pH was higher for WPSB versus EPSBM. The Plenish diets decreased the prevalence of Ruminococcus and increased that of Eubacterium and Treponema in whole ruminal contents. Total-tract apparent digestibility of organic matter and crude protein were decreased by WPSB compared with ESBM and EPSBM. Compared with the other treatments, urinary N excretion was increased by EPSBM and fecal N excretion was greater for WPSB. Treatments had marked effects on milk fatty acid profile. Generally, the Plenish diets increased mono-unsaturated (mostly cis-9 18:1) and decreased polyunsaturated, total trans-, and conjugated linoleic fatty acids concentrations in milk fat. In this study, compared with conventional, high-linoleic-acid variety extruded soybean meal, the Plenish soybean diets increased milk fat concentration and tended to increase fat yield, decreased feed efficiency, and modified milk fatty acid profile in a manner expected from the greater concentration of oleic acid in Plenish soybean oil.

Short communication: Preference for flavored concentrate premixes by dairy cows.

Flavor preferences may be used to stimulate feed intake in dairy cows, which may improve use of robotic milking systems and increase feed intake of sick cows. A cafeteria-design experiment was used to determine if dairy cows have flavor preferences. Sixteen lactating Holstein cows averaging 197±32d in milk, 1.9±0.8 lactations, 27.8±4.2kg/d of dry matter intake, and 41.5±7.4kg/d of milk yield were involved in the experiment. Cows were offered 7 flavored concentrate premixes (FCP) and 1 control premix. The FCP flavors were anise, fenugreek, honey, orange, thyme, molasses, and vanilla; the absence of flavor, neutral, acted as a control. The inclusion rate of the flavors in FCP was 250 to 300g/t on an as-is basis. Cows were not adapted to the flavors before the experiment. Cows were housed in a tiestall barn and offered, on each day, 4 different FCP (1kg each) in plastic bins placed in front of each cow. The experiment lasted 6 consecutive days. Each FCP was presented to each cow once every 2d, 2h after the morning feeding. Flavors and position of the bins in front of the cows were randomized. As a result, each flavor was presented to each cow 3 times during the experiment, at 3 different bin locations. Each cow had access to the FCP for 5min from the time they started eating. Eating time and amount eaten were recorded. The vanilla and fenugreek FCP were consumed the most, at 408 and 371g/5-min offering, respectively, whereas the orange and anise FCP were consumed the least, at 264 and 239g/5-min offering, respectively. Similarly, cows spent the most time eating the vanilla and fenugreek FCP at 99 and 75 s/offering, respectively, and the least amount of time eating the orange and anise FCP at 49 and 50 s/offering, respectively. We detected an effect of bin position: the 2 center FCP were consumed more than the outer 2 FCP. Flavor had no effect on consumption rate. In conclusion, relative to the control, concentrate intake was not affected by flavor, but dairy cows may prefer vanilla or fenugreek flavors when offered a novel choice.

Short communication: Comparison of the GreenFeed system with the sulfur hexafluoride tracer technique for measuring enteric methane emissions from dairy cows.

The objective of this study was to compare 2 commonly used techniques for measuring methane emissions from ruminant animals: the GreenFeed (GF) system and the sulfur hexafluoride (SF6) technique. The study was part of a larger experiment in which a methane inhibitor, 3-nitrooxypropanol, fed at 4 application rates (0, 40, 60, and 80 mg/kg of feed dry matter) decreased enteric methane emission by an average of 30% (measured by both GF and SF6) in a 12-wk experiment with 48 lactating Holstein cows fed a total mixed ration. The larger experiment used a randomized block design and was conducted in 2 phases (February to May, phase 1, and June to August, phase 2), with 2 sets of 24 cows in each phase. Using both GF and SF6 techniques, methane emission data were collected simultaneously during experimental wk 2, 6, and 12 (phase 1) and 2, 9, and 12 (phase 2), which corresponded to a total of 6 sampling periods. During each sampling period, 8 spot samples of gas emissions (staggered over a 3-d period) were collected from each cow using GF, as well as 3×24-h collections using the SF6 technique. Methane emission data were averaged per cow for the statistical analysis. The mean methane emission was 373 (standard deviation=96.3) and 405 (standard deviation=156) g/cow per day for GF and SF6, respectively. Coefficients of variation for the 2 methods were 25.8 and 38.6%, respectively; correlation and concordance between the 2 methods were 0.40 and 0.34, respectively. The difference in methane emission between the 2 methods (SF6 - GF) within treatment was from 46 to 144 and 24 to 27 g/d for phases 1 and 2, respectively. In the conditions of this experiment, the SF6 technique produced larger variability in methane emissions than the GF method. The overall difference between the 2 methods was on average about 8%, but was not consistent over time, likely influenced by barn ventilation and background methane and SF6 concentrations.

Effects of rumen-protected methionine, lysine, and histidine on lactation performance of dairy cows.

The objective of this study was to evaluate the effects of supplementing a metabolizable protein (MP)-deficient diet with rumen-protected (RP) Met, Lys, and His, individually or combined, on the performance of lactating dairy cows. The experiment was a 9-wk randomized complete block design with 72 Holstein cows. Following a 2-wk covariate period, cows were blocked by days in milk, milk yield, and parity, and randomly assigned to 1 of the following 6 treatments: (1) MP-adequate diet [MPA; +243g/d MP balance, according to the National Research Council (2001) requirements]; (2) MP-deficient diet (MPD; -54g/d MP balance); (3) MPD supplemented with RPMet (MPDM); (4) MPD supplemented with RPLys (MPDL); (5) MPD supplemented with RPHis (MPDH); and (6) MPD supplemented with RPMet, RPLys, and RPHis (MPDMLH). Dry matter intake (DMI), yields of milk and milk components (fat, protein, lactose) and energy-corrected milk (ECM), feed and ECM feed efficiencies, and milk and plasma urea N were decreased by MPD, compared with MPA. Supplementation of the MPD diet with RPLys increased milk protein content and plasma glucose concentration and tended to increase milk urea N. Addition of RPHis tended to increase DMI, increased milk protein concentration, and numerically increased yields of milk fat, protein, and ECM. In addition to the trends for increased DMI and milk fat content, and higher milk protein concentration, supplementation of the 3 RP AA also increased yields of milk fat, protein, and ECM and ECM feed efficiency. Relative to MPA, milk N efficiency tended to be increased by MPD. Concentrations of plasma essential AA (except Met and Thr) were decreased by MPD compared with MPA. Supplementation of RPMet, RPLys, and RPHis increased plasma Met (except for MPDM), Lys, and His concentrations, respectively. Cows fed MPD had lower blood hemoglobin concentration and numerically higher plasma ghrelin than cows fed MPA. Concentration of total saturated fatty acids in milk fat were or tended to be higher for MPD compared with MPA and MPDMLH, respectively. Concentration of total polyunsaturated and yield of milk odd- and branched-chain fatty acids were or tended to be decreased by MPD compared with MPA. Overall, the results of this study confirm our previous data and suggest that His stimulates DMI and the combination of the 3 RP AA (Met, Lys, and His) has the potential to improve milk and milk component yields in dairy cows fed MP-deficient diets.

Effect of 3-nitrooxypropanol on methane and hydrogen emissions, methane isotopic signature, and ruminal fermentation in dairy cows.

The objective of this crossover experiment was to investigate the effect of a methane inhibitor, 3-nitrooxypropanol (3NOP), on enteric methane emission, methane isotopic composition, and rumen fermentation and microbial profile in lactating dairy cows. The experiment involved 6 ruminally cannulated late-lactation Holstein cows assigned to 2 treatments: control and 3NOP (60 mg/kg of feed dry matter). Compared with the control, 3NOP decreased methane emission by 31% and increased hydrogen emission from undetectable to 1.33 g/d. Methane emissions per kilogram of dry matter intake and milk yield were also decreased 34% by 3NOP. Milk production and composition were not affected by 3NOP, except milk fat concentration was increased compared with the control. Concentrations of total VFA and propionate in ruminal fluid were not affected by treatment, but acetate concentration tended to be lower and acetate-to-propionate ratio was lower for 3NOP compared with the control. The 3NOP decreased the molar proportion of acetate and increase those of propionate, butyrate, valerate, and isovalerate. Deuterium-to-hydrogen ratios of methane and the abundance of (13)CH3D were similar between treatments. Compared with the control, minor (4‰) depletion in the (13)C/(12)C ratio was observed for 3NOP. Genus composition of methanogenic archaea (Methanobrevibacter, Methanosphaera, and Methanomicrobium) was not affected by 3NOP, but the proportion of methanogens in the total cell counts tended to be decreased by 3NOP. Prevotella spp., the predominant bacterial genus in ruminal contents in this experiment, was also not affected by 3NOP. Compared with the control, Ruminococcus and Clostridium spp. were decreased and Butyrivibrio spp. was increased by 3NOP. This experiment demonstrated that a substantial inhibition of enteric methane emission by 3NOP in dairy cows was accompanied with increased hydrogen emission and decreased acetate-to-propionate ratio; however, neither an effect on rumen archaeal community composition nor a significant change in the isotope composition of methane was observed.

Identification of roles for the SNARE-associated protein, SNAP29, in mouse platelets.

Platelets are critical for maintaining vascular hemostasis, but also play a major role in the formation of occlusive cardiovascular and cerebrovascular thrombi under disease conditions. Secretion of platelet alpha and dense granules is a requirement for efficient thrombus formation. Understanding and targeting the mechanisms of secretion is important to aid the development of effective antithrombotics. SNAP29 is a tSNARE found in platelets, but whose role has not been defined. Using a platelet-specific SNAP29 knockout mouse model, we assessed the role of SNAP29 in platelet secretion and function under standardized conditions and also in in vitro and in vivo thrombosis. The data showed no major defects in SNAP29-null platelets, but revealed a minor defect in α-granule secretion and a significant increase in embolization rate of thrombi in vivo. These data suggest that SNAP29 contributes to the regulation of platelet α-granule secretion and thrombus stability, possibly partially masked by functional redundancy with other tSNAREs, such as SNAP23.

Leukemia-associated Rho guanine-nucleotide exchange factor is not critical for RhoA regulation, yet is important for platelet activation and thrombosis in mice.

BACKGROUND: RhoA is an important regulator of platelet responses downstream of Gα13 , yet we still know little about its regulation in platelets. Leukemia-associated Rho guanine-nucleotide exchange factor (GEF [LARG]), a RhoA GEF, is highly expressed in platelets and may constitute a major upstream activator of RhoA. To this end, it is important to determine the role of LARG in platelet function and thrombosis. METHODS AND RESULTS: Using a platelet-specific gene knockout, we show that the absence of LARG results in a marked reduction in aggregation and dense-granule secretion in response to the thromboxane mimetic U46619 and proteinase-activated receptor 4-activating peptide, AYPGKF, but not to adenosine diphosphate. In a ferric chloride thrombosis model in vivo, this translated into a defect, under mild injury conditions. Importantly, agonist-induced RhoA activation was not affected by the absence of LARG, although basal activity was reduced, suggesting that LARG may play a housekeeper role in regulating constitutive RhoA activity. CONCLUSIONS: LARG plays an important role in platelet function and thrombosis in vivo. However, although LARG may have a role in regulating the resting activation state of RhoA, its role in regulating platelet function may principally be through RhoA-independent pathways, possibly through other Rho family members.

Chloride channels are necessary for full platelet phosphatidylserine exposure and procoagulant activity.

Platelets enhance thrombin generation at sites of vascular injury by exposing phosphatidylserine during necrosis-like cell death. Anoctamin 6 (Ano6) is required for Ca(2+)-dependent phosphatidylserine exposure and is defective in patients with Scott syndrome, a rare bleeding disorder. Ano6 may also form Cl(-) channels, though the role of Cl(-) fluxes in platelet procoagulant activity has not been explored. We found that Cl(-) channel blockers or removal of extracellular Cl(-) inhibited agonist-induced phosphatidylserine exposure. However, this was not due to direct inhibition of Ca(2+)-dependent scrambling since Ca(2+) ionophore-induced phosphatidylserine exposure was normal. This implies that the role of Ano6 in Ca(2+-)dependent PS exposure is likely to differ from any putative function of Ano6 as a Cl(-) channel. Instead, Cl(-) channel blockade inhibited agonist-induced Ca(2+) entry. Importantly, Cl(-) channel blockers also prevented agonist-induced membrane hyperpolarization, resulting in depolarization. We propose that Cl(-) entry through Cl(-) channels is required for this hyperpolarization, maintaining the driving force for Ca(2+) entry and triggering full phosphatidylserine exposure. This demonstrates a novel role for Cl(-) channels in controlling platelet death and procoagulant activity.