Separate offering of forages and concentrates to lactating dairy cows: Effects on lactational performance, enteric methane emission, and efficiency of nutrient utilization.

The objective was to evaluate the effects of separate offering of feed ingredients (SF) and frequency of concentrate feeding versus offering a TMR, on lactational performance, ruminal fermentation, enteric CH4 emissions, nutrient digestibility, N use efficiency, milk fatty acid profile, and blood variables in mid-lactation dairy cows. Twenty-four Holstein cows (12 primi- and 12 multiparous) averaging (±SD) 141 ± 35 DIM and 43 ± 6 kg/d of milk yield (MY) at the beginning of the study were used in a replicated 3 × 3 Latin square design experiment with 3 periods of 28 d each, composed of 7 d for adaptation to the diets, 11 d for estimation of net energy and metabolizable protein requirements, and 10 d for data and samples collection. Cows were grouped based on parity, DIM, and MY into 4 Latin squares. Treatment allocation was balanced for carryover effects, and cows within square were assigned to (1) basal diet fed ad libitum as TMR; (2) basal diet fed as SF with forages fed ad libitum and concentrates fed 3×/d (SF×3); or (3) basal diet fed as SF with forages fed ad libitum and concentrates fed 6×/d (SF×6). Compared with TMR, SF decreased total DMI by 1.2 kg/d. Treatments did not affect MY, milk components, or ECM yield, except for a decrease in milk fat concentration and an increase in milk urea N by SF×3, compared with TMR. Feed efficiency (kg of MY/kg of DMI) was increased by 7% in SF, compared with TMR. Ruminal molar proportion of acetate and acetate-to-propionate ratio were decreased, whereas molar proportion of propionate was increased by SF×3, compared with TMR and SF×6. There was a 9% decrease in daily CH4 production by SF, compared with TMR. Enteric CH4 yield (per kg of DMI) was not affected by treatments in the current study. Methane intensity per kilogram of MY tended to be decreased by 10% in SF, compared with TMR. The sums of odd- and branched-chain, odd-chain, and anteiso milk fatty acids tended to be or were increased by SF, compared with TMR. Intake of nutrients tended to be or were decreased by SF, compared with TMR. The digestibility of amylase-treated NDF tended to be decreased and ADF digestibility was decreased by 3% in SF, compared with TMR. Urinary and fecal N excretions were not affected by treatments. As a percentage of total N intake, separate offering of feed ingredients increased milk N secretion, indicating an increased N use efficiency by SF, compared with TMR. Blood total fatty acid concentration was decreased by SF relative to TMR. Compared with both TMR and SF×6, SF×3 increased blood urea N concentration. Overall, feed and N use efficiencies were increased by separate offering of feed ingredients, and increasing the frequency of concentrate feeding promoted ruminal fermentation effects similar to those obtained by feeding a TMR.

Lactational performance, ruminal fermentation, and enteric gas emission of dairy cows fed an amylase-enabled corn silage in diets with different starch concentrations.

This study investigated the effects of feeding an amylase-enabled corn silage (ACS) on the performance and enteric gas emissions in lactating dairy cows. Following a 2-wk covariate period, 48 mid-lactation Holstein cows were assigned to 1 of 3 treatments in a 10-wk randomized complete block design experiment. Treatments were diets containing the same proportion of corn silage (40% of dietary DM) as follows: (1) a conventional hybrid corn silage control (CON), (2) ACS replacing the control silage (ADR), and (3) the ADR diet replacing soybean hulls with ground corn grain to achieve the same dietary starch concentration as CON (ASR). Control corn silage and ACS were harvested on the same day and contained 40.3% and 37.1% DM and (% of DM): 37.2% and 41.0% NDF and 37.1% and 30.0% starch, respectively. Enteric gas emissions were measured using the GreenFeed system. Two cows were culled due to health-related issues during the covariate period. Ruminal fluid was collected from 24 cows (8 per treatment) using the orogastric ruminal sampling technique. When compared with CON, cows fed ADR had increased DMI during experimental wk 3, 4, and 9, but treatment did not affect milk or ECM milk yields (39.0 kg/d on average; SEM = 0.89). Compared with CON, feed efficiency (per unit of milk, but not ECM) tended to be lower for ADR, whereas milk true protein concentration (a tendency) and yield were lower for ASR. Milk urea N was decreased by both ADR and ASR diets relative to CON. Compared with CON, daily CH4 emission and emission intensity were increased by ADR but not ASR. Total protozoal count tended to be increased by both diets formulated with ACS when compared with control corn silage. Total-tract digestibility of dietary NDF was greater for ASR, and that of ADF was greater for both ADR and ASR versus CON. The molar proportion of acetate (a tendency) and acetate-to-propionate ratio were increased by ADR, but not ASR, when compared with CON. Replacement of CON with ACS (having lower starch concentration) in the diet of dairy cows increased DMI during the initial weeks of the experiment, maintained ECM, tended to decrease feed efficiency, and increased enteric CH4 emissions, likely due to increased intake of digestible fiber, compared with CON.

A meta-analysis of methane-mitigation potential of feed additives evaluated in vitro.

A systematic literature review of in vitro studies was performed to identify methane (CH4) mitigation interventions with a potential to reduce CH4 emission in vivo. Data from 277 peer-reviewed studies published between 1979 and 2018 were reviewed. Individual CH4 mitigation interventions were classified into 14 categories of feed additives based on their type, chemical composition, and mode of action. Response variables evaluated were absolute CH4 emission (number of treatment means comparisons = 1,325); total volatile fatty acids (n = 1,007), acetate (n = 783), propionate (n = 792), and butyrate (n = 776) concentrations; acetate to propionate ratio (n = 675); digestibility of dry matter (n = 489), organic matter (n = 277), and neutral detergent fiber (n = 177). Total gas production was used as an explanatory variable in the model for CH4 production. Relative mean difference between treatment and control means reported in the studies was calculated and used for statistical analysis. The robust variance estimation method was used to analyze the effects of CH4 mitigation interventions. In vitro CH4 production was decreased by antibodies (-38.9%), chemical inhibitors (-29.2%), electron sinks (-18.9%), essential oils (-18.2%), plant extracts (-14.5%), plant inclusion (-11.7%), saponins (-14.8%), and tannins (-14.5%). Overall effects of direct-fed microbials, enzymes, macroalgae, and organic acids supplementation did not affect CH4 production in the current meta-analysis. When considering the effects of individual mitigation interventions containing a minimum number of 4 degrees of freedom within feed additives categories, Enterococcus spp. (i.e., direct-fed microbial), nitrophenol (i.e., electron sink), and Leucaena spp. (i.e., tannins) decreased CH4 production by 20.3%, 27.1%, and 23.5%, respectively, without extensively, or only slightly, affecting ruminal fermentation and digestibility of nutrients. It should be noted, however, that although the total number of publications (n = 277) and treatment means comparisons (n = 1,325 for CH4 production) in the current analysis were high, data for most mitigation interventions were obtained from less than 5 observations (e.g., maximum number of observations was 4, 7, and 22 for nitrophenol, Enterococcus spp., and Leucaena spp., respectively), because of limited data available in the literature. These should be further evaluated in vitro and in vivo to determine their true potential to decrease enteric CH4 production, yield, and intensity. Some mitigation interventions (e.g., magnesium, Heracleum spp., nitroglycerin, ß-cyclodextrin, Leptospermum pattersoni, Fructulus Ligustri, Salix caprea, and Sesbania grandiflora) decreased in vitro CH4 production by over 50% but did not have enough observations in the database. These should be more extensively investigated in vitro, and the dose effect must be considered before adoption of mitigation interventions in vivo.

Lactational performance, enteric methane emission, and nutrient utilization of dairy cows supplemented with botanicals.

The objective of this study was to evaluate lactational performance, enteric gas emissions, ruminal fermentation, nutrient use efficiency, milk fatty acid profile, and energy and inflammatory markers in blood of peak-lactation dairy cows fed diets supplemented with Capsicum oleoresin or a combination of Capsicum oleoresin and clove oil. A 10-wk randomized complete block design experiment was conducted with 18 primiparous and 30 multiparous Holstein cows. Cows were blocked based on parity, days in milk, and milk yield (MY), and randomly assigned to 1 of 3 treatments (16 cows/treatment): (1) basal diet (CON); (2) basal diet supplemented with 300 mg/cow per day of Capsicum oleoresin (CAP); and (3) basal diet supplemented with 300 mg/cow per day of a combination of Capsicum oleoresin and clove oil (CAPCO). Premixes containing ground corn (CON), CAP, or CAPCO were mixed daily with the basal diet at 0.8% of dry matter intake (DMI). Supplementation of the diet with CAP or CAPCO did not affect DMI, MY, milk components, and feed efficiency of the cows. Body weight (BW) was increased during the last 2 wk of the experiment by CAP and CAPCO, compared with CON. The botanicals improved BW gain (0.85 and 0.66 kg/d for CAP and CAPCO, respectively, compared with -0.01 kg/d for CON) and CAP enhanced the efficiency of energy utilization, compared with CON (94.5% vs. 78.4%, respectively). Daily CH4 emission was not affected by treatments, but CH4 emission yield (per kg of DMI) and intensity (per kg of MY) were decreased by up to 11% by CAPCO supplementation, compared with CON and CAP. A treatment × parity interaction indicated that the CH4 mitigation effect was pronounced in primiparous but not in multiparous cows. Ruminal molar proportion of propionate was decreased by botanicals, compared with CON. Concentrations of trans-10 C18:1 and total trans fatty acids in milk fat were decreased by CAP and tended to be decreased by CAPCO, compared with CON. Total-tract apparent digestibility of nutrients was not affected by treatments, except for a tendency for decreased starch digestibility in cows supplemented with botanicals. Blood concentrations of ß-hydroxybutyrate, total fatty acids, and insulin were not affected by botanicals. Blood haptoglobin concentration was increased by CAP in multiparous but not in primiparous cows. Lactational performance of peak-lactation dairy cows was not affected by the botanicals in this study, but they appeared to improve efficiency of energy utilization and partitioned energy toward BW gain. In addition, CH4 yield and intensity were decreased in primiparous cows fed CAPCO, suggesting a potential positive environmental effect of the combination of Capsicum oleoresin and clove oil supplementation.

A meta-analysis of methane mitigation potential of feed additives evaluated in vitro.

A systematic literature review of in vitro studies was performed to identify methane (CH4) mitigation interventions with a potential to reduce CH4 emission in vivo. Data from 277 peer-reviewed studies published between 1979 and 2018 were reviewed. Individual CH4 mitigation interventions were classified into 14 categories of feed additives based on their type, chemical composition, and mode of action. Response variables evaluated were absolute CH4 emission (number of treatment means comparisons = 1,325); total volatile fatty acids (VFA; n = 1,007), acetate (n = 783), propionate (n = 792), and butyrate (n = 776) concentrations; acetate to propionate ratio (A:P; n = 675); digestibility of dry matter (DM; n = 489), organic matter (OM; n = 277), and neutral detergent fiber (NDF; n = 177). Total gas production was used as an explanatory variable in the model for CH4 production. Relative mean difference between treatment and control means reported in the studies were calculated and used for statistical analysis. Robust variance estimation method was used to analyze the effects of CH4 mitigation interventions. In vitro CH4 production was decreased by antibodies (-38.9%), chemical inhibitors (-29.2%), electron sinks (-18.9%), essential oils (-18.2%), plant extracts (-14.5%), plants inclusion (-11.7%), saponins (-14.8%), and tannins (-14.5%). Overall effects of direct fed microbials, enzymes, macroalgae, and organic acids supplementation did not affect CH4 production in the current meta-analysis. When considering the effects of individual mitigation interventions containing a minimum number of 4 degrees of freedom within feed additives categories, Enterococcus spp. (i.e., direct fed microbial), nitrophenol (i.e., electron sink), and Leucaena spp. (i.e., tannins) decreased CH4 production by 20.3, 27.1, and 23.5%, respectively, without extensively, or only slightly, affecting ruminal fermentation and digestibility of nutrients. It should be noted, however, that although the total number of publications (n = 277) and treatment means comparisons (n = 1,325 for CH4 production) in the current analysis were high, data for most mitigation interventions were obtained from less than 5 observations (e.g., maximum number of observations was 4, 7, and 22 for nitrophenol, Enterococcus spp., and Leucaena spp., respectively), because of limited data available in the literature. These should be further evaluated in vitro and in vivo to determine their true potential to decrease enteric CH4 production, yield, and intensity. Some mitigation interventions (e.g., magnesium, Heracleum spp., nitroglycerin, ß-cyclodextrin, Leptospermum pattersoni, Fructulus Ligustri, Salix caprea, and Sesbania grandiflora) decreased in vitro CH4 production by over 50% but did not have enough observations in the database. These should be more extensively investigated in vitro, and the dose effect must be considered before adoption of mitigation interventions in vivo.

Protein requirements for pregnant dairy cows.

We aimed to estimate the protein requirements of pregnant Holstein × Gyr cows. A total of 61 Holstein × Gyr (HG) cows were used, with an average initial body weight (BW) of 480 ± 10.1 kg and age 5 ± 0.5 yr. Cows were divided into 3 groups: pregnant (n = 43), nonpregnant (n = 12), and baseline (n = 6). Baseline animals were slaughtered before starting the experiment to estimate the initial body composition of the remaining animals. Pregnant and nonpregnant cows received 2 diets: maintenance and ad libitum. Pregnant cows were slaughtered at 139, 199, 241, and 268 gestation days (GD). First, we used data only from nonpregnant cows to determine the requirements for maintenance and growth in adult cows. The requirements of metabolizable protein for maintenance (MPm; grams of empty metabolic BW [EBW0.75] per day) were estimated using a linear regression between the metabolizable protein intake (MPI, g/d) and average daily gain (g/d), and the MPm was defined as the intercept divided by the average metabolic BW. Net protein requirements for gain (NPg; g/d) were estimated by the first derivative of the allometric equation between final crude protein in the body (kg) and the final empty BW (EBW; kg). The efficiency of use of metabolizable protein for gain (k) was calculated from the regression between the retained protein (g EBW0.75/d) and the MPI (g EBW0.75/d), and k was the slope of this regression. The MPI was estimated by combining microbial protein synthesis (purine derivatives) with the digestible rumen undegradable protein [(total protein intake - rumen-degradable protein) × intestinal digestibility]. Second, an exponential model was used to fit the protein accumulation in the gestational components in the function of GD. The first derivative of that model was considered the net requirement for gestation (NPgest). The efficiency of protein utilization for gestation (kgest) was calculated by the iterative method using the equation Δ = MPI - (MPm + NPg/kg + NPgest/kgest), where kg is efficiency of protein utilization for gain. The iteration was performed aiming at a zero deviation between observed MPI and metabolizable protein (MP) estimated by the requirements determined herein. We obtained a value of 3.88 g EBW0.75/d for MPm. The estimation of NPg can be calculated according to the following equation: NPg = 0.716 × (EBWopen-0.308) × EBGc, where EBWopen is the EBW (kg) for nonpregnant animals and EBGc is the empty body gain (kg/d) corrected for the gestational component. The k was determined as 0.347. The NPgest requirements were determined as NPgest (g/d) = 0.0008722 × exp(0.01784 × GD) × (calf weight/35). The kgest was 0.625. It is important to highlight that different methods of MP estimates should not be mixed and that the proposed method requires the estimation of microbial protein (estimated via urinary estimates), which might limit practical application. In conclusion, new studies should be conducted to validate our results and the methodology adopted to determine protein requirements for pregnancy in dairy cows. Due to the pattern of protein accumulation in the gestational components, we suggest an exponential model to describe protein requirements for pregnancy for dairy cows.

Lactational performance, rumen fermentation, nutrient use efficiency, enteric methane emissions, and manure greenhouse gas-emitting potential in dairy cows fed a blend of essential oils.

The objective of this experiment was to investigate the effects of an essential oil (EO) blend on lactational performance, rumen fermentation, nutrient utilization, blood variables, enteric methane emissions and manure greenhouse gas-emitting potential in dairy cows. A randomized complete block design experiment was conducted with 26 primiparous and 22 multiparous Holstein cows. A 2-wk covariate and a 2-wk adaptation periods preceded a 10-wk experimental period used for data and sample collection. Treatments were: (1) basal diet supplemented with placebo (CON); and (2) basal diet supplemented with a blend of EO containing eugenol and geranyl acetate as main compounds. Supplementation with EO did not affect dry matter intake, milk and energy-corrected milk yields, and feed efficiency of cows, compared with CON. Milk fat and lactose concentrations were increased, and milk total solids (TS) concentration and milk fat yield tended to be increased by EO. Multiparous cows supplemented with EO tended to have slightly decreased dry matter and crude protein digestibility compared with CON multiparous cows. There was a tendency for increased ruminal pH by EO, whereas other rumen fermentation variables did not differ between treatments. Daily methane emission was not affected by EO supplementation, but methane emission intensity per kg of milk fat was decreased by 8.5% by EO. Methane emission intensity per kg of milk lactose and milk TS were decreased and methane emission intensity per kg of milk yield tended to be decreased by up to 10% in EO multiparous cows, but not in primiparous cows. The greenhouse gas-emitting potential of manure was not affected by EO supplementation. Compared with CON, fecal nitrogen excretion was increased by EO supplementation in multiparous, but not in primiparous cows, and milk nitrogen secretion (as a % of nitrogen intake) tended to be increased in EO supplemented cows. Blood variables were not affected by EO supplementation in the current study. Overall, dietary supplementation of EO did not affect lactational performance of the cows, although milk fat and lactose concentrations were increased. Most enteric methane emission metrics were not affected, but EO decreased methane intensity per kg of milk fat by 8.5%, compared with the control.

Production effects of extruded soybean meal replacing canola meal in the diet of lactating dairy cows.

This study investigated the effects of extruded soybean meal (ESBM) in comparison with canola meal (CM) fed on an equivalent crude protein (CP) basis on lactational performance and ruminal fermentation of dairy cows. Following a 2-wk covariate period, 48 Holstein cows averaging (±SD): 146 ± 46 d in milk (DIM) and 43 ± 7 kg/d milk yield (MY) were assigned 1 of 2 treatment diets in a randomized complete block design experiment, which included a 2-wk period for dietary treatment adaptation before experimental data were collected. Following the adaptation period, samples and experimental data were collected for a total of 7 wk. Cows were blocked based on parity, DIM, and MY. Treatment diets contained 15.8% CM (containing 41.2% CP) or 13.2% ESBM (with 48.7% CP) of total mixed ration dry matter (DM), with similar inclusion of other feed ingredients. The CM diet was supplemented with canola oil, whereas the ESBM diet was supplemented with soybean hulls to achieve similar ether extract and neutral detergent fiber contents between the diets. Urea and rumen-protected Met and Lys were added to both diets to meet or exceed cow recommendations. Whole-ruminal digesta samples were collected from 10 (5 per treatment) ruminally cannulated cows. Eight cannulated cows were removed during the last week of the experiment to participate in another study. Treatment did not affect DM intake and MY or energy-corrected MY of the cows. Energy-corrected MY, apart from experimental wk 5, was similar between treatments. Apart from experimental wk 3 and 7, milk fat concentration and yield were greater for cows fed ESBM compared with CM. In multiparous cows only, milk true protein yield was greater for cows fed CM compared with ESBM. Ruminal concentration of total volatile fatty acids and the molar proportion of acetate were greater for ESBM, and propionate and valerate were greater in cows fed CM. Acetate to propionate ratio was greater for cows fed ESBM versus CM diet. Compared with the CM diet, the ESBM diet increased plasma concentrations of Ile, Leu, and Phe but not the sum of essential AA. Apparent total-tract digestibility of acid detergent fiber was greater in cows fed ESBM relative to CM. In this experiment, CM and ESBM included on an equal CP basis in the diet of dairy cows, resulted in similar DM intake, MY, and feed efficiency.

Effects of a combination of Capsicum oleoresin and clove essential oil on metabolic status, lactational performance, and enteric methane emissions in dairy cows.

Botanical extracts have a potential to modify ruminal fermentation while enhancing metabolism and immunity in dairy cows. The objective of this study was to investigate the effects of a combination of Capsicum oleoresin and clove essential oil (botanicals; BTC) on lactational performance, nutrient utilization, enteric methane (CH4) emissions, and blood parameters in dairy cows. Twenty Holstein cows (12 multiparous and 8 primiparous) averaging (±SD) 77 ± 28 d in milk in the beginning of the study were used in a replicated 4 × 4 Latin square design experiment with 4 periods of 28 d each. Cows were grouped into squares based on parity, milk yield and days in milk, and assigned to 1 of 4 treatments: control (CON), 150, 300, or 600 mg/cow per day of BTC. Cows received the same basal diet and BTC were top-dressed on the total mixed ration once daily. Dry matter intake, milk production, and milk composition were not affected by BTC supplementation, except for milk fat content that tended to be increased in BTC, compared with CON. Daily CH4 emission (measured using the GreenFeed system) was linearly decreased by up to 7.5% with increasing doses of BTC. Treatment decreased CH4 yield (kg of CH4 ÷ kg of DMI) and tended to decrease CH4 intensity (kg of CH4 ÷ kg of milk or energy-corrected milk yields) by 5% in BTC, compared with CON. Supplementation of BTC resulted in a quadratic decrease of serum ß-hydroxybutyrate in all cows, and a linear decrease of serum insulin concentration in primiparous but not in multiparous cows. Nutrient utilization and other blood parameters (e.g., blood cells count) were not affected by BTC in the current study. The reduction of enteric CH4 emission demonstrates a moderate mitigation effect on carbon footprint of milk by BTC supplementation. These results must be further investigated and confirmed in longer-term experiments.

Performance of dairy cows fed normal- or reduced-starch diets supplemented with an exogenous enzyme preparation.

The objective of this study was to investigate the effects of supplementation of an exogenous enzyme preparation (EEP) on performance, total-tract digestibility of nutrients, plasma AA profile, and milk fatty acids composition in lactating dairy cows fed a reduced-starch diet compared with a normal-starch diet (i.e., positive control). Forty-eight Holstein cows (28 primiparous and 20 multiparous) were enrolled in a 10-wk randomized complete block design experiment with 16 cows per treatment. Treatments were as follows: (1) normal-starch diet (control) containing (% dry matter basis) 24.8% starch and 33.0% neutral detergent fiber (NDF), (2) reduced-starch diet (RSD) containing 18.4% starch and 39.1% NDF, or (3) RSD supplemented with 10 g/cow per day of an EEP (ENZ). The EEP contained amylolytic and fibrolytic activities and was top-dressed on the total mixed ration at the time of feeding. Compared with normal-starch diet, dry matter intake and milk and energy-corrected milk (ECM) yields were lower (on average by 7.1, 9.5, and 7.2%, respectively) for cows on the RSD treatments. Concentrations, but not yields, of milk fat and total solids were increased by RSD. Energy-corrected milk feed efficiency did not differ among treatments. Total-tract digestibility of NDF tended to increase by RSD treatments. Plasma AA concentrations were not affected by treatment, except that of 3-methylhistidine was increased by ENZ, compared with RSD. Blood glucose concentration tended to be lower in cows on the RSD treatments, but ENZ increased glucose and tended to increase insulin concentrations at 4 h after feeding when compared with RSD. Cows on the RSD treatments had decreased concentrations of de novo fatty acids and tended to have increased concentrations of preformed fatty acids in milk. Overall, decreasing dietary starch concentration by 26% decreased dry matter intake, milk, and ECM yields, but ECM feed efficiency was not different among treatments. The negative effects of reducing dietary starch on production were not attenuated by the EEP.

Production effects and bioavailability of N-acetyl-l-methionine in lactating dairy cows.

Two experiments were conducted to investigate the production effects of N-acetyl-l-methionine (NALM; experiment 1) and to estimate its bioavailability (BA) and rumen escape (RE; experiment 2), respectively, in lactating dairy cows. In experiment 1, 18 multiparous Holstein cows were used in a replicated, 3 × 3 Latin square design experiment with three 28-d periods. Treatments were (1) basal diet estimated to supply 45 g/d digestible Met (dMet) or 1.47% of metabolizable protein (MP; control), (2) basal diet top-dressed with 32 g/d of NALM to achieve dMet supply of 2.2% of MP, and (3) basal diet top-dressed with 56 g/d of NALM to achieve dMet supply of 2.6% of MP. The NALM treatments supplied estimated 17 and 29 g/d dMet from NALM, respectively, based on manufacturer's specifications. In experiment 2, 4 rumen-cannulated lactating Holstein cows were used in a 4 × 4 Latin square design experiment with four 12-d periods. A 12-d period for baseline data collection and 4 d for determination of RE of NALM preceded the Latin square experiment. For determination of RE, 30 g of NALM were dosed into the rumen simultaneously with Cr-EDTA (used as a rumen fluid kinetics marker) and samples of ruminal contents were collected at 0 (before dosing), 1, 2, 4, 6, 8, 10, 14, 18, and 24 h after dosing. Rumen escape of NALM was calculated using the estimated passage rate based on the measured Cr rate of disappearance. Bioavailability of abomasally dosed NALM was determined using the area under the curve of plasma Met concentration technique. Two doses of l-Met (providing 7.5 and 15 g of dMet) and 2 doses of NALM (11.2 and 14.4 g of dMet) were separately pulse-dosed into the abomasum of the cows and blood was collected from the jugular vein for Met concentration analysis at 0 (before dosing), 1, 2, 4, 6, 8, 10, 12, 14, 18, and 24 h after dosing. Supplementation of NALM did not affect DMI, milk yield, feed efficiency, or milk protein and lactose concentrations and yields in experiment 1. Milk fat concentration and energy-corrected milk yield decreased linearly with NALM dose. Plasma Met concentration was not affected by NALM dose. The estimated relative BA of abomasally dosed NALM (experiment 2) was 50% when dosed at 14.4 g/cow (11.2 g/d dMet from NALM) and 24% when dosed at 28.8 g/cow (14.4 g/d dMet from NALM). The estimated RE of NALM was 19% based on the measured kp of Cr at 11%/h. The total availability of ingested NALM was estimated at 9.5% for the lower NALM dose when taking into account RE (19%) and bioavailability in the small intestine (50%). Overall, NALM supplementation to mid-lactation dairy cows fed a MP-adequate basal diet below NRC (2001) recommendations (45 g/d or 1.47% Met of MP) decreased milk fat and energy-corrected milk yields but did not affect milk or milk true protein yields. Further evaluation of BA of NALM at different doses is warranted. In addition, intestinal conversion of NALM to Met needs additional investigation to establish a possible saturation of the enzyme aminoacylase I at higher NALM doses.

Lactational performance and plasma and muscle amino acid concentrations in dairy cows fed diets supplying 2 levels of digestible histidine and metabolizable protein.

The objective of this experiment was to investigate the effect of dietary levels of digestible histidine (dHis) and MP on lactational performance and plasma and muscle concentrations of free AA in dairy cows. A randomized block design experiment was conducted with 48 Holstein cows, including 20 primiparous, averaging (±SD) 103 ± 22 d in milk and 45 ± 9 kg/d milk yield at the beginning of the experiment. A 2-wk covariate period preceded 12 experimental wk, of which 10 wk were for data and sample collection. Experimental treatments were (1) MP-adequate (MPA) diet with 2.1% dHis of MP (MPA2.1), (2) MPA with 3.0% dHis (MPA3.0), (3) MP-deficient (MPD) diet with 2.1% dHis (MPD2.1), and (4) MPD with 3.0% dHis (MPD3.0). Actual dHis supply was estimated at 64, 97, 57, and 88 g/d, respectively. Diets supplied MP at 110% (MPA) and 96% (MPD) of NRC 2001 dairy model requirements calculated based on DMI and production data during the experiment. Dry matter intake and milk yield data were collected daily, milk samples for composition and blood samples for AA analysis were collected every other week, and muscle biopsies at the end of covariate period, and during wk 12 of the experiment. The overall DMI was not affected by dHis or MP level. Milk yield tended to be increased by 3.0% dHis compared with 2.1% dHis. Milk true protein concentration and yield were not affected by treatments, whereas milk urea nitrogen concentration was lower for MPD versus the MPA diet. Milk fat concentration was lower for MPD versus MPA. There was a MP × dHis interaction for milk fat yield and energy-corrected milk; milk fat was lower for MPD3.0 versus MPD2.1, but similar for cows fed the MPA diet regardless of dHis level whereas energy-corrected milk was greater for MPA3.0 versus MPA2.1 but tended to be lower for MPD3.0 versus MPD2.1. Plasma His concentration was greater for cows fed dHis3.0, and concentration of sum of essential AA was greater, whereas carnosine, 1-Methyl-His and 3-Methyl-His concentrations were lower for cows fed MPA versus MPD diet. Muscle concentration of His was greater for cows fed dHis3.0 treatment. The apparent efficiency of His utilization was increased at lower MP and His levels. Overall, cows fed a corn silage-based diet supplying MP at 110% of NRC (2001) requirements tended to have increased ECM yield and similar milk protein yield to cows fed a diet supplying MP at 96% of requirements. Supplying dHis at 3.0% of MP (or 86 and 96 g/d, for MPD3.0 and MPA3.0, respectively) tended to increase milk yield and increased plasma and muscle concentrations of His but had minor or no effects on other production variables in dairy cows.

Efficacy of antibiotic prophylaxis in intraoral bone grafting procedures: a systematic review and meta-analysis.

The purpose of this systematic review was to investigate the efficacy of antibiotic prophylaxis (AP) in intraoral bone grafting procedures for the prevention of postoperative infection (POI). Electronic and manual searches were conducted to identify randomized controlled trials (RCTs). The primary outcome assessed was receptor site POI; secondary outcomes assessed included donor site POI, wound dehiscence, pain, graft failure, need for re-grafting, adverse events, patient satisfaction, and quality of life. A random-effects meta-analysis was conducted to obtain risk ratios of dichotomous data. Four RCTs were selected: one examined AP versus placebo and concluded that there was an increased risk of POI without AP; three examined comparative antibiotic regimens and found no statistically significant difference between them. A meta-analysis of prophylactic regimens, including data from the two RCTs that compared preoperative AP to perioperative AP, indicated no statistically significant difference in POI outcomes (P= 0.94, risk ratio 0.94). It was not possible to conduct further meta-analyses for POIs or for any secondary outcomes due to insufficient published data. The risk of bias assessment indicated an overall unclear risk of bias. On the basis of the present review, there is insufficient evidence to support or refute AP for the prevention of POIs in intraoral bone graft placement procedures.

A novel marker for assessment of liver matrix remodeling: an enzyme-linked immunosorbent assay (ELISA) detecting a MMP generated type I collagen neo-epitope (C1M).

A competitive enzyme-linked immunosorbent assay (ELISA) for detection of a type I collagen fragment generated by matrix metalloproteinases (MMP) -2, -9 and -13, was developed (CO1-764 or C1M). The biomarker was evaluated in two preclinical rat models of liver fibrosis: bile duct ligation (BDL) and carbon tetra chloride (CCL4)-treated rats. The assay was further evaluated in a clinical study of prostate-, lung- and breast-cancer patients stratified according to skeletal metastases. A technically robust ELISA assay specific for a MMP-2, -9 and -13 neo-epitope was produced and seen to be statistically elevated in BDL rats compared to baseline levels as well as significantly elevated in CCL4 rats stratified according to the amount of total collagen in the livers. CO1-764 levels also correlated significantly with total liver collagen and type I collagen mRNA expression in the livers. Finally, the CO1-764 marker was not correlated with skeletal involvement or number of bone metastases. This ELISA has the potential to assess the degree of liver fibrosis in a non-invasive manner.

Recycling carbon fibre reinforced polymers for structural applications: technology review and market outlook.

Both environmental and economic factors have driven the development of recycling routes for the increasing amount of carbon fibre reinforced polymer (CFRP) waste generated. This paper presents a review of the current status and outlook of CFRP recycling operations, focusing on state-of-the-art fibre reclamation and re-manufacturing processes, and on the commercialisation and potential applications of recycled products. It is shown that several recycling and re-manufacturing processes are reaching a mature stage, with implementations at commercial scales in operation, production of recycled CFRPs having competitive structural performances, and demonstrator components having been manufactured. The major challenges for the sound establishment of a CFRP recycling industry and the development of markets for the recyclates are summarised; the potential for introducing recycled CFRPs in structural components is discussed, and likely promising applications are investigated.

Advances in osteoclast biology resulting from the study of osteopetrotic mutations.

Osteopetrosis is the result of mutations affecting osteoclast function. Careful analyses of osteopetrosis have provided instrumental information on bone remodeling, including the coupling of bone formation to bone resorption. Based on a range of novel genetic mutations and the resulting osteoclast phenotypes, we discuss how osteopetrosis models have clarified the function of the coupling of bone formation to bone resorption, and the pivotal role of the osteoclast and their function in this phenomenon. We highlight the distinct possibility that osteoclast activities can be divided into two separate avenues: bone resorption and control of bone formation.

Reversion of mtDNA depletion in a patient with TK2 deficiency.

Mutations in the thymidine kinase 2 (TK2) gene cause a myopathic form of the mitochondrial DNA depletion syndrome (MDS). Here, the authors report the unusual clinical, biochemical, and molecular findings in a 14-year-old patient in whom pathogenic mutations were identified in the TK2 gene. This report extends the phenotypic expression of primary TK2 deficiency and suggests that factors other than TK2 may modify expression of the clinical phenotype in patients with MDS syndrome.

Effects of taurine on polymorphonuclear phagocytosis activity in burned patients.

This study determines the effects of taurine (Tau) on phagocytosis of polymorphonuclear neutrophils (PMN) isolated from normal subjects (n = 41) and severely burned patients (n = 20). Phagocytosis was measured by nitroblue of tetrazolium (NBT) reduction in samples with and without latex bead stimulation. Taurine was added at doses of 0.2, 0.4, 0.8 and 1.6 mM to stimulated samples. In control cells there were statistically significant increases in phagocytosis after addition of Tau 0.8 mM and 1.6 mM to as compared to samples without Tau addition (295 +/- 23% and 330 +/- 35% vs. 248 +/- 18%; mean +/- S.E.; p < 0.05). A statistically significant increase in phagocytosis was observed in cells from the burned population after addition of Tau 1.6 mM (288 +/- 38% vs. 198 +/- 13%; mean +/- S.E.; p < 0.05). No changes in phagocytosis were found in cells from a subgroup of burn patients (n = 13) followed over 7, 15 and 21 days. These results indicate that taurine supplementation in vitro at doses of 0.8 to 1.6 mM improves the phagocytic capacity of neutrophils in healthy subjects and in patients with severe burn injury, mainly when neutrophil function is unaltered.

Role of putrescine in cell proliferation in a colon carcinoma cell line.

OBJECTIVES: Putrescine, the precursor for higher polyamine biosynthesis, is necessary for cell growth in mammals. Ornithine decarboxylase (ODC) activity and polyamine production are increased in neoplastic cells. Using colon cancer cell line derived from a tumor with high metastatic potential (CT-26), our objective was to study the effect of exogenous putrescine on ODC regulation, polyamine metabolism, and cell proliferation. METHODS: Cells cultured with fetal calf serum were exposed to 100, 550, and 1000 microM putrescine for 24 h. RESULTS: Intracellular free putrescine, determined by high-performance liquid chromatography, showed a statistically significant increase in exposed cells compared with controls and a significant correlation with levels of the metabolite present in the medium (r = 0.93; P < 0.001). Bromodeoxyuridine incorporation into newly synthesized DNA, a marker of cell proliferation, showed a statistically significant increase in the three putrescine groups as opposed to the control group. In samples with added aminoguanidine, significant increases in DNA synthesis were observed in the 550- and 1000-microM putrescine groups as opposed to the control group. Spermidine and spermine intracellular contents in all three putrescine-treated groups remained below control levels. No statistical differences in ODC enzymatic activity or ODC mRNA content were observed. Newly incorporated putrescine stimulated colon tumor cell growth. CONCLUSIONS: Because neither enhanced conversion into the higher polyamines nor aminoguanidine inhibition of proliferation was observed, we suggest that this effect can be attributed to the putrescine molecule itself.