Immunostimulating activity of Uncaria tomentosa in RAW 264.7 macrophages.

Uncaria tomentosa is a plant native to the Amazon that has immunomodulatory and antitumor properties due to the alkaloids found in the plant, being able to modify the immune response by potentiating or suspending the action of cytokines secreted by macrophages that induce the immune response, either by the classical route (M1) or through the alternative route (M2). Macrophages activated by M1 convert L-arginine into L-citrulline and nitric oxide (NO), whereas macrophages activated by the M2 pathway use the enzymatic activity of arginase to convert the same substrate into L-ornithine and urea. The aim of this work was to evaluate the immunostimulating activity of the crude hydroalcoholic extract from the bark of the U. tomentosa stem in RAW 264.7 macrophages. Concentrations of 0.2, 0.1 and 0.05 mg/mL of U. tomentosa extract associated with LPS, INF-γ and IL-4 inducers were tested by determining NO production and arginase enzyme activity. Nitric oxide production was enhanced by the extract when associated with LPS and LPS + INF-γ inducers. In the activity of the arginase enzyme, the extract decreased the stimulation of IL-4 on the enzyme, mainly at 0.2 mg/mL concentration. Therefore, it is concluded that the crude hydroalcoholic extract of the stem bark of U. tomentosa in RAW 264.7 cells, at a concentration of 0.2 mg/mL, showed considerable pro-inflammatory activity.

Replacing ground corn with soyhulls plus palmitic acid in low metabolizable protein diets with or without rumen-protected amino acids: Effects on production and nutrient utilization in dairy cows.

We previously observed that diets with reduced starch concentration decreased yields of milk and milk protein in dairy cows fed low metabolizable protein diets. Supplementation of reduced-starch diets with a lipid source may attenuate or eliminate production losses. Our objective was to investigate the effects of partially replacing ground corn with soyhulls plus a palmitic acid-enriched supplement on dry matter (DM) intake, milk yield and composition, plasma AA concentration, and N and energy utilization in cows fed low metabolizable protein diets (mean = -68 g/d balance) with or without rumen-protected Met, Lys, and His (RP-MLH). Sixteen multiparous Holstein cows averaging (mean ± standard deviation) 112 ± 28 d in milk, 724 ± 44 kg of body weight, and 46 ± 5 kg/d of milk in the beginning of the study were used in a replicated 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. Each period lasted 21 d, consisting of 14 d for diet adaptation and 7 d for data and sample collection. Diets were fed as follows: (1) high starch (HS), (2) HS plus RP-MLH (HS+AA), (3) reduced starch plus a palmitic acid-enriched supplement (RSPA), and (4) RSPA plus RP-MLH (RSPA+AA). The HS diet contained (DM basis) 26% ground corn and 7% soyhulls, and the RSPA diet had 10% ground corn, 22% soyhulls, and 1.5% palmitic acid. The HS diet averaged (DM basis) 32.6% starch and 4% ether extract, while starch and ether extract concentrations of the RSPA diet were 21.7 and 5.9%, respectively. All 4 diets had (DM basis) 40% corn silage, 5% mixed-mostly grass haylage, 5% grass hay, and 50% concentrate. Diets did not affect DM intake and milk yield. Contrarily, feeding RSPA and RSPA+AA increased yields of energy-corrected milk (47.0 vs. 44.8 kg/d) and milk fat (1.65 vs. 1.50 kg/d) compared with HS and HS+AA. Milk fat concentration tended to decrease when RP-MLH was supplemented to HS, but no change was seen when added to RS (starch level × RP-MLH interaction). Milk and plasma urea N increased, and milk N efficiency decreased in cows fed RSPA and RSPA+AA versus HS and HS+AA. Apparent total-tract digestibilites of crude protein and neutral detergent fiber, as well as urinary urea N and total N excretion, were greater in cows offered RSPA and RSPA+AA than HS and HS+AA. Plasma Met and His concentrations increased with supplemental RP-MLH. Intake of gross energy and digestible energy and the output of urinary and milk energy were all greater with feeding RSPA and RSPA+AA versus HS and HS+AA. In summary, partially replacing ground corn with soyhulls plus palmitic acid in diets supplemented or not with RP-MLH increased milk fat yield and fiber digestibility and maintained DM intake and milk yield, but with decreased milk N efficiency and elevated urinary N excretion.

Supplementation of Ascophyllum nodosum meal and monensin: Effects on diversity and relative abundance of ruminal bacterial taxa and the metabolism of iodine and arsenic in lactating dairy cows.

Previous research has shown that the brown seaweed Ascophyllum nodosum (ASCO) has antimicrobial and antioxidant properties and also increases milk I concentration. We aimed to investigate the effects of supplementing ASCO meal or monensin (MON) on ruminal fermentation, diversity and relative abundance of ruminal bacterial taxa, metabolism of I and As, and blood concentrations of thyroid hormones, antioxidant enzymes, and cortisol in lactating dairy cows. Five multiparous ruminally cannulated Jersey cows averaging (mean ± standard deviation) 102 ± 15 d in milk and 450 ± 33 kg of body weight at the beginning of the study were used in a Latin square design with 28-d periods (21 d for diet adaptation and 7 d for data and sample collection). Cows were fed ad libitum a basal diet containing (dry matter basis) 65% forage as haylage and corn silage and 35% concentrate and were randomly assigned to 1 of the following 5 dietary treatments: 0, 57, 113, or 170 g/d of ASCO meal, or 300 mg/d of MON. Supplements were placed directly into the rumen once daily after the morning feeding. Diets had no effect on ruminal pH and NH3-N concentration, which averaged 6.02 and 6.86 mg/dL, respectively. Total volatile fatty acid concentration decreased linearly in cows fed incremental amounts of ASCO meal. Supplementation with ASCO meal did not change the ruminal molar proportions of volatile fatty acids apart from butyrate, which responded quadratically with the lowest values observed at 56 and 113 g/d of ASCO supplementation. Compared with the control diet or diets containing ASCO meal, cows fed MON showed greater molar proportion of propionate. Diets did not affect the α diversity indices Shannon, Simpson, and Fisher for ruminal bacteria. However, feeding incremental levels of ASCO meal linearly decreased the relative abundance of Tenericutes in ruminal fluid. Monensin increased the relative abundance of the CAG:352 bacterial genus in ruminal fluid compared with the control diet. Linear increases in response to ASCO meal supplementation were observed for the concentrations and output of I in serum, milk, urine, and feces. Fecal excretion of As increased linearly in cows fed varying amounts of ASCO meal, but ASCO did not affect the concentration and secretion of As in milk. The plasma activities of the antioxidant enzymes and the serum concentrations of thyroid hormones did not change. In contrast, circulating cortisol decreased linearly in diets containing ASCO meal. The apparent total-tract digestibilities of dry matter, organic matter, and crude protein increased linearly with ASCO meal, but those of neutral and acid detergent fiber were not affected. In summary, feeding incremental amounts of ASCO meal decreased serum cortisol concentration, and increased I concentrations and output in serum, milk, feces, and urine.

Dietary starch level and rumen-protected methionine, lysine, and histidine: Effects on milk yield, nitrogen, and energy utilization in dairy cows fed diets low in metabolizable protein.

Our objective was to investigate the interactions between starch level and rumen-protected Met, Lys, His (RP-MLH) on milk yield, plasma AA concentration, and nutrient utilization in dairy cows fed low metabolizable protein diets (mean = -119 g/d of metabolizable protein balance). Sixteen multiparous Holstein cows (138 ± 46 d in milk, 46 ± 6 kg/d in milk) were used in a replicated 4 × 4 Latin square with a 2 × 2 factorial arrangement of treatments. Each period lasted 21 d with 14 d for diet adaptation and 7 d for data and sample collection. Dietary starch level varied by replacing (dry matter basis) pelleted beet pulp and soyhulls with ground corn resulting in the following treatments: (1) 20% pelleted beet pulp and 10% soyhulls (reduced starch = RS), (2) RS plus RP-MLH (RS+AA), (3) 30% ground corn (high starch = HS), and (4) HS plus RP-MLH (HS+AA). Dietary starch concentrations averaged 12.3 and 34.4% for RS and HS basal diets, respectively. Diets were supplemented with RP-MLH products to supply digestible Met, Lys, and His. Compared with RS and RS+AA diets, HS and HS+AA diets increased yields of milk (37.9 vs. 40.1 kg/d) and milk protein (1.07 vs. 1.16 kg/d) and decreased dry matter intake (25.9 vs. 25.2 kg/d), milk urea N (12.6 vs. 11.0 mg/dL), and plasma urea N (13.3 vs. 11.6 mg/dL). Milk N efficiency was greater in cows fed the HS and HS+AA than RS and RS+AA diets (28.9 vs. 25%), and RP-MLH supplementation improved milk true protein concentration. Starch level × RP-MLH interactions were observed for plasma concentrations of Arg and Lys, with RP-MLH being more effective to increase plasma Arg (+16%) and Lys (+23%) when supplemented to the RS than the HS basal diet. Replacing pelleted beet pulp and soyhulls with ground corn lowered the plasma concentrations of all essential AA except Met and Thr. In addition, the plasma concentrations of His and Met increased with RP-MLH. The apparent total-tract digestibilities of neutral and acid detergent fiber were lower, and those of starch and ether extract greater in cows offered the HS and HS+AA diets than RS and RS+AA diets. Urinary excretion of urea N decreased by replacing pelleted beet pulp and soyhulls with ground corn. Enteric CH4 production, CH4 yield, and CH4 intensity all decreased in the HS and HS+AA versus RS and RS+AA diets. Diets did not affect the intakes of gross energy, metabolizable energy, and net energy of lactation. In contrast, digestible energy intake increased with feeding the RS and RS+AA diets, whereas CH4 energy decreased in cows fed the HS and HS+AA diets. Supplementation with RP-MLH had no effect on energy utilization variables. Overall, the lack of interactions between dietary starch level and RP-MLH supplementation on most variables measured herein showed that the effects of starch intake and RP-MLH were independent or additive.

The MAPKinase Signaling and the Stimulatory Protein-1 (Sp1) Transcription Factor Are Involved in the Phototherapy Effect on Cytokines Secretion from Human Bronchial Epithelial Cells Stimulated with Cigarette Smoke Extract.

The present study was aimed to investigate the phototherapy effect with low-level laser on human bronchial epithelial cells activated by cigarette smoke extract (CSE). Phototherapy has been reported to actuate positively for controlling the generation/release of anti-inflammatory and pro-inflammatory mediators from different cellular type activated by distinct stimuli. It is not known whether the IL-8 and IL-10 release from CSE-stimulated human bronchial epithelium (BEAS) cells can be influenced by phototherapy. Human bronchial epithelial cell (BEAS) line was cultured in a medium with CSE and irradiated (660 nm) at 9 J. Apoptosis index was standardized with Annexin V and the cellular viability was evaluated by MTT. IL-8, IL-10, cAMP, and NF-κB were measured by ELISA as well as the Sp1, JNK, ERK1/2, and p38MAPK. Phototherapy effect was studied in the presence of mithramycin or the inhibitors of JNK or ERK. The IL-8, cAMP, NF-κB, JNK, p38, and ERK1/2 were downregulated by phototherapy. Both the JNK and the ERK inhibitors potentiated the phototherapy effect on IL-8 as well as on cAMP secretion from BEAS. On the contrary, IL-10 and Sp1 were upregulated by phototherapy. The mithramycin blocked the phototherapy effect on IL-10. The results suggest that phototherapy has a dual effect on BEAS cells because it downregulates the IL-8 secretion by interfering with CSE-mediated signaling pathways, and oppositely upregulates the IL-10 secretion through of Sp1 transcription factor. The manuscript provides evidence that the phototherapy can interfere with MAPK signaling via cAMP in order to attenuate the IL-8 secretion from CSE-stimulated BEAS. In addition, the present study showed that phototherapy effect is driven to downregulation of the both the IL-8 and the ROS secretion and at the same time the upregulation of IL-10 secretion. Besides it, the increase of Sp-1 transcription factor was crucial for laser effect in upregulating the IL-10 secretion. The dexamethasone corticoid produces a significant inhibitory effect on IL-8 as well as ROS secretion, but on the other hand, the corticoid blocked the IL-10 secretion. Taking it into consideration, it is reasonable to suggest that the beneficial effect of laser therapy on lung diseases involves its action on unbalance between pro-inflammatory and anti-inflammatory mediators secreted by human bronchial epithelial cells through different signaling pathway.

Assessing the potential of milk iodine intake to mitigate iodine deficiency in pregnant women of the United States via supplementation of Ascophyllum nodosum meal to dairy cows: A sensitivity analysis.

The brown seaweed Ascophyllum nodosum is known to bioaccumulate iodine (I). Previous research showed a linear relationship between A. nodosum meal (ASCO) intake and milk I concentration in dairy cows. Thus, improving milk I concentration by supplementation of ASCO to dairy cows may be a compelling strategy to naturally boost I intake in humans. A sensitivity analysis was conducted to gain insights regarding how different milk I intakes affect the I status of pregnant women relative to the United States Institute of Medicine (US IOM) recommended dietary allowance (RDA; 220 µg/d) and the World Health Organization (WHO) recommended nutrient intake (RNI; 250 µg/d) for I. Four studies in which dairy cows received various amounts of ASCO generated the milk I data set used in the sensitivity analysis. The annual per capita consumption of 2% reduced-fat milk in the United States, converted to daily intake (i.e., 0.26 cup; 1 cup = 236.6 mL), was used as the actual milk intake in the simulations. Five additional milk intake scenarios (2, 3, 4, and 5 times the actual per capita milk consumption and the 3 cups-equivalent recommended by the 2015-2020 Dietary Guidelines for American were also included in the sensitivity analysis with varying milk I concentrations (180, 765, and 483 µg/L). The 180, 765, and 483 µg/L values are milk I concentrations derived from cows not receiving ASCO or fed various amounts of ASCO in the diet or a single level (113 g/d) during the grazing season, respectively. With the actual United States milk per capita consumption of 0.26 cup/d and milk I concentrations of 180, 765, and 483 µg/L, 5.09, 21.7, and 13.6% of the RDA for I for pregnant women were met based on the US IOM, respectively. Similarly, 4.48, 19, and 12% of the RNI for I advised by the WHO was achieved with intake of 0.26 cup/d of milk containing I concentrations of 180, 765, and 483 µg/L, respectively. When 3 cups/d was included in the simulations, 58.2, 247, and 156% (US IOM), and 51.2, 217, and 137% (WHO) of the RDA or RNI for I required by gestating women was satisfied with milk I concentrations of 180, 765, and 483 µg/L, respectively. A regression analysis between I intake and milk I concentration revealed that 103 g/d of ASCO in the diet of dairy cows reached the maximum 500 µg/L threshold of I in milk recommended by the European Food Society Authority. Overall, milk from dairy cows fed ASCO can prevent I deficiency in pregnant women, but the amount of ASCO fed to cows needs to be fine-tuned to avoid excess I in milk. Further research is required to better understand the interactions between goitrogenic compounds from forages and concentrates and milk I concentration in cows fed ASCO. Research to evaluate the concentration of I in retail organic milk should be also conducted because of the high prevalence of ASCO fed in organic dairies in the United States.

Liquid molasses interacts with buffers to affect ruminal fermentation, milk fatty acid profile, and milk fat synthesis in dairy cows fed high-concentrate diets.

We aimed to evaluate the effects of feeding sugarcane liquid molasses (LM) with or without a commercial buffer mix (BFM) on ruminal fermentation parameters, milk fatty acid (FA) profile, and milk yield and composition in dairy cows fed high-concentrate diets (35:65 forage-to-concentrate ratio). Eight multiparous Holstein cows (4 ruminally cannulated) averaging 165 ± 12 d in milk at the beginning of the study were randomly assigned to a replicated 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. Each period lasted 21 d with 14 d for diet adaptation and 7 d for data and sample collection. Cows were fed the following diets: (1) no LM or BFM supplementation (CTRL), (2) LM without BFM supplementation (MOL), (3) BFM without LM supplementation (BUF), and (4) LM plus BFM supplementation (COMBO). These 4 isonitrogenous and isoenergetic diets were formulated by replacing (dry matter basis) 5% ground corn with LM, whereas BFM replaced wheat bran at 0.8% of the diet. Significant LM × BFM interactions were observed for the duration of ruminal pH below 5.8, molar proportion of propionate, acetate-to-propionate ratio, milk proportions of trans-10 18:1 and total trans FA, and concentration and yield of milk fat. Feeding MOL and BUF alone were effective on reducing the time that ruminal pH remained below 5.8 compared with the CTRL treatment, and the COMBO diet decreased it further. A similar pattern was observed for the ruminal molar proportion of propionate. The milk proportions of trans-10 18:1 and total trans FA dropped significantly with BFM or LM supplementation versus cows fed CTRL, and the COMBO diet decreased these variables further. Note, however, that these changes elicited by the COMBO diet were not in the same magnitude as those caused by MOL or BUF fed alone. The ruminal molar proportion of acetate increased with the BUF diet and that of butyrate increased in cows fed MOL, but mean ruminal pH was not affected by treatments. Diets with LM resulted in increased concentrations of short- and medium-chain FA in milk fat. The yield of 3.5% fat-corrected milk increased significantly in cows fed MOL or BUF due to the improved concentration of milk fat. A trend and a significant increase for energy-corrected milk were observed with feeding MOL or BUF, respectively. Overall, inclusion of LM and BFM appears to reduce milk trans-10 18:1 FA and total trans FA by modulating ruminal pH and volatile FA profile in cows fed high-concentrate diets.

Production and nitrogen metabolism in lactating dairy cows fed finely ground field pea plus soybean meal or canola meal with or without rumen-protected methionine supplementation.

We showed previously that dairy cows fed [diet dry matter (DM) basis] 25% finely ground field pea (GFP) plus rumen-protected (RP)-Met and RP-Lys had greater milk true protein yield and plasma Met concentration, but lower plasma His, compared with those fed GFP without rumen-protected AA supplementation. The goal of the present study was to investigate the effects of diets containing soybean meal (SBM) or canola meal (CM) with or without a source of RP-Met on production, nutrient digestibility, and N metabolism in cows fed 25% GFP. Sixteen mid-lactation Holstein cows were used in a replicated 4 × 4 Latin square with a 2 × 2 factorial arrangement of treatments (21-d periods). Cows were fed (DM basis) 35% corn silage, 14% grass-legume haylage, 25% GFP, and 11% SBM or 13.5% CM with or without 0.095% RP-Met supplementation as Smartamine M (Adisseo USA Inc., Alpharetta, GA). Protein source effects were observed for most production variables; cows fed diets containing CM plus GFP had greater DM intake, yields of milk and milk fat and true protein, and milk N efficiency than those offered SBM plus GFP. Feeding CM plus GFP also reduced the concentrations of milk urea N and plasma urea N, and the urinary excretion of NH3 N and urea N, suggesting improved N use efficiency. Moreover, replacing SBM with CM increased the concentrations of all essential AA (except Arg) in plasma collected from the coccygeal blood vessels. A protein source × RP-Met interaction was observed for the concentration of His in coccygeal plasma, with circulating His decreasing only when RP-Met was supplemented to the diet containing SBM plus GFP. Based on the arteriovenous difference method, Lys was the first limiting AA overall, with Met being the first limiting AA in diets that did not receive RP-Met. Apparent total-tract digestibilities of DM, organic matter, N, and acid detergent fiber increased with feeding SBM plus GFP versus CM plus GFP. Most variables were not affected by RP-Met supplementation, except plasma Met concentration, which increased by 63%. Collectively, our results indicate that CM appears to be a better companion rumen-degradable protein source to GFP than SBM, due to improved yields of milk and milk protein and N use efficiency.

Production, milk iodine, and nutrient utilization in Jersey cows supplemented with the brown seaweed Ascophyllum nodosum (kelp meal) during the grazing season.

Kelp meal (KM) is a supplement made from the brown seaweed Ascophyllum nodosum, known to bioaccumulate iodine (I) and to be the richest source of phlorotannins, which can inhibit ruminal proteolysis and microbial growth. The objective of this study was to investigate the effects of KM on production, milk I, concentrations of blood metabolites, apparent total-tract digestibility of nutrients, and CH4 emissions in grazing dairy cows. Eight multiparous Jersey cows averaging (mean ± SD) 175 ± 60 d in milk and 12 primiparous Jersey cows averaging 142 ± 47 d in milk at the beginning of the study were assigned to either 0 g/d of KM (control diet, CTRL) or 113 g/d of KM (brown seaweed diet, BSW) in a randomized complete block design. Diets were formulated to yield a 70:30 forage-to-concentrate ratio and consisted of (dry matter basis): 48% cool-season perennial herbage and 52% partial TMR (pTMR). Each experimental period (n = 3) lasted 28 d, with data and sample collection taking place during the last 7 d of each period. Cows had approximately 16.5 h of access to pasture daily. Herbage dry matter intake increased, and total dry matter intake tended to increase in cows fed BSW versus the CTRL diet. Milk yield and concentrations and yields of milk components were not affected by diets. Similarly, blood concentrations of cortisol, glucose, fatty acids, and thyroxine did not change with feeding CTRL or BSW. However, a diet × period interaction was observed for milk I concentration; cows offered the BSW diet had greater milk I concentration during periods 1, 2, and 3, but the largest difference between BSW and CTRL was observed in period 2 (579 vs. 111 µg/L, respectively). Except for period 2, the concentration of milk I in cows fed KM did not exceed the 500 µg/L threshold recommended for human consumption. Diet × period interactions were also found for serum triiodothyronine concentration, total-tract digestibilities of crude protein and acid detergent fiber, CH4 production, and urinary excretion of purine derivatives. Overall, the lack of KM effects on milk yield and concentrations and yields of milk components indicate that dairy producers should consider costs before making KM supplementation decisions during the grazing season. Future research is needed to evaluate the concentration of I in retail organic milk because of the high prevalence of KM supplementation in northeastern and midwestern US organic dairies and possibly in other regions of the country.

Incremental amounts of rumen-protected histidine increase plasma and muscle histidine concentrations and milk protein yield in dairy cows fed a metabolizable protein-deficient diet.

The dairy industry can benefit from low crude protein (CP) diets due to reduced N excretion, but shortages of Met, Lys, and His may limit milk protein synthesis. We studied the effect of incremental amounts of rumen-protected (RP)-His on plasma and muscle AA profile, nutrient utilization, and yields of milk and milk true protein in dairy cows. Eight multiparous Holstein cows (130 ± 30 d in milk) were randomly assigned to treatment sequences in a replicated 4 × 4 Latin square design with 28-d experimental periods. Treatments included a basal diet composed (dry matter basis) of 50% corn silage, 15% haylage, and 35% concentrate supplemented with 0, 82, 164, and 246 g/d of RP-His and 11 g/d of RP-Met. Milk, plasma, and muscle samples were collected weekly or every other week during all 4 periods, whereas spot urine and fecal grab samples were taken only in wk 4 of each period. Data were analyzed individually by week using linear, quadratic, and cubic orthogonal polynomials and repeated measures. Plasma His increased linearly with RP-His during wk 1 (30.3 to 57.2 µM) to wk 4 (33.2 to 63.1 µM). Plasma carnosine increased linearly with supplemental RP-His except in wk 2. No treatment effect was observed for plasma 3-methylhistidine except a quadratic effect in wk 3. Inclusion of RP-His showed linear effects on muscle His in wk 2 (20.1 to 32.5 µM) and 4 (20.3 to 35.5 µM). Whereas muscle anserine and carnosine concentrations were not affected by treatments in wk 4, anserine responded quadratically and carnosine showed a trend for a quadratic response to RP-His in wk 2. During wk 4, treatments did not affect urinary excretion of total purine derivatives, as well as dry matter intake and milk concentrations of fat and true protein. In contrast, milk yield tended to increase linearly (31.2 to 32.7 kg/d) and milk true protein yield responded linearly (0.93 to 0.98 kg/d) and tended to increase quadratically to RP-His supplementation in wk 4. Also, milk urea-N (11.7 to 12.9 mg/dL) and urinary excretion of urea-N (23.7 to 27.0% of N intake) increased linearly with feeding RP-His in wk 4. Overall, RP-His was effective to enhance plasma and muscle concentrations of His and milk protein synthesis. Elevated milk urea-N and urinary excretion of urea-N suggest that plasma His may have exceeded the requirement with excess N converted to urea in the liver. Future research is needed to determine the bioavailability of RP-His supplements to improve the accuracy of diet formulation for AA.

Production, milk fatty acid profile, and nutrient utilization in grazing dairy cows supplemented with ground flaxseed.

Flaxseed has been extensively used as a supplement for dairy cows because of its high concentrations of energy and the n-3 fatty acid (FA) cis-9,cis-12,cis-15 18:3. However, limited information is available regarding the effect of ground flaxseed on dry matter intake (DMI), ruminal fermentation, and nutrient utilization in grazing dairy cows. Twenty multiparous Jersey cows averaging (mean ± standard deviation) 111 ± 49 d in milk in the beginning of the study were used in a randomized complete block design to investigate the effects of supplementing herbage (i.e., grazed forage) with ground corn-soybean meal mix (control diet = CTRL) or ground flaxseed (flaxseed diet = FLX) on animal production, milk FA, ruminal metabolism, and nutrient digestibility. The study was conducted from June to September 2013, with data and sample collection taking place on wk 4, 8, 12, and 16. Cows were fed a diet formulated to yield a 60:40 forage-to-concentrate ratio consisting of (dry matter basis): 40% cool-season perennial herbage, 50% partial total mixed ration, and 10% of ground corn-soybean meal mix or 10% ground flaxseed. However, estimated herbage DMI averaged 5.59 kg/d or 34% of the total DMI. Significant treatment by week interactions were observed for milk and blood urea N, and several milk FA (e.g., trans-10 18:1). No significant differences between treatments were observed for herbage and total DMI, milk yield, feed efficiency, concentrations and yields of milk components, and urinary excretion of purine derivatives. Total-tract digestibility of organic matter decreased, whereas that of neutral detergent fiber increased with feeding FLX versus CTRL. No treatment effects were observed for ruminal concentrations of total volatile FA and NH3-N, and ruminal proportions of acetate and propionate. Ruminal butyrate tended to decrease, and the acetate-to-propionate ratio decreased in the FLX diet. Most saturated and unsaturated FA in milk fat were changed. Specifically, milk proportion of cis-9,cis-12,cis-15 18:3, Σn-3 FA, and Σ18C FA increased, whereas that of cis-9,cis-12 18:2, Σn-6 FA, Σ odd-chain FA, Σ<16C FA, and Σ16C FA decreased with feeding FLX versus the CTRL diet. In conclusion, feeding FLX did not change yields of milk and milk components, but increased milk n-3 FA. Therefore, costs and industry adoption of premiums for n-3-enriched milk will determine the adoption of ground flaxseed in pasture-based dairy farms.

Production, milk and plasma fatty acid profile, and nutrient utilization in Jersey cows fed flaxseed oil and corn grain with different particle size.

We aimed to compare the effects of ground (GC) or cracked corn (CC), with or without flaxseed oil (FSO), on milk yield, milk and plasma fatty acid (FA) profile, and nutrient digestibility in Jersey cows fed diets formulated to contain similar starch concentrations. Twelve multiparous organic-certified Jersey cows averaging (mean ± standard deviation) 455 ± 41.9 kg of body weight and 152 ± 34 d in milk and 4 primiparous organic-certified Jersey cows averaging (mean ± standard deviation) 356 ± 2.41 kg of body weight and 174 ± 30 d in milk in the beginning of the experiment were used. Cows were randomly assigned to treatment sequences in a replicated 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. Each period lasted 24 d with 18 d for diet adaptation and 6 d for data and sample collection. Treatments were fed as total mixed rations consisting of (dry matter basis): (1) 0% FSO + 27.1% GC, (2) 0% FSO + 28.3% CC, (3) 3% FSO + 27.1% GC, and (4) 3% FSO + 28.3% CC. All cows were offered 55% of the total diet dry matter as mixed grass-legume baleage and treatments averaged 20% starch. Significant FSO × corn grain particle size interactions were observed for some variables including milk concentration of lactose and proportions of cis-9,cis-12,cis-15 18:3 in milk and plasma. The proportion of cis-9,cis-12,cis-15 18:3 in milk and plasma decreased slightly when comparing GC versus CC in 0% FSO cows, but a larger reduction was observed in 3% FSO cows. Dry matter intake did not differ and averaged 16.1 kg/d across diets. Feeding 3% FSO increased yields of milk and milk fat and lactose and feed and milk N efficiencies, but decreased fat, true protein, and MUN concentrations and apparent total-tract digestibility of fiber. The Σ branched-chain, Σ<16C, Σ16C, and Σn-6 FA decreased, whereas Σ18C, Σcis-18:1, and Σtrans-18:1 FA increased in 3% versus 0% FSO cows. No effect of corn particle size was observed for production and milk components. However, the apparent total-tract digestibility of starch was greater in GC than CC cows. Compared with CC, GC increased Σ branched-chain, Σ<16C, Σ16C, Σn-6 FA, and decreased Σ18C and Σ cis-18:1 FA in milk fat. Overall, results of this study are more directly applicable to dairy cows fed low starch, mixed grass-legume baleage-based diets.

The impact of direct-fed microbials and enzymes on the health and performance of dairy cows with emphasis on colostrum quality and serum immunoglobulin concentrations in calves.

Thirty-six cows were blocked by calving date and randomly assigned to one of three treatments. Cows were on treatments 3 weeks prepartum through 8 weeks post-partum. Treatments were as follows: (i) no direct-fed microbial (DFM) or cellulase and amylase enzymes (C), (ii) 45.4 g/day of DFM (D) or (iii) 45.4 g/day of DFM and 18.2 g/day of enzyme (DE). Total mixed ration fed and refused were measured daily to determine dry matter intake (DMI). Blood samples were taken three times weekly and analysed for ß-hydroxybutyrate, glucose and non-esterified fatty acids. Body weight (BW) was measured weekly. Colostrum was weighed and analysed for IgA and IgG concentration. Calves were fed 4 L of colostrum within 2 hr of birth. Calf blood samples were taken at 0 and 24 hr for analysis of IgA and IgG concentrations and apparent efficiency of absorption. Milk yield was measured daily and samples collected weekly. Initial BW was different among treatments with D being lesser than C or DE treatments. Body weight, weight gain, efficiency of gain, DMI and blood parameters were unaffected. Treatment did not affect colostrum yield. Ash percentage of colostrum tended to increase with D and DE, while IgA and total solids yield decreased with D. Colostrum fat yield was decreased in D and DE. Treatments did not impact BW, serum IgA and IgG concentrations or apparent efficiency of absorption of calves. Post-partum BW, DMI, blood parameters, milk production and composition were unaffected by treatment. However, cows on D gained more BW and tended to have greater efficiency of gain compared to those on DE, but were similar to C. Somatic cell scores were greatest for D. Results indicate that DFM and enzyme supplementation did not improve health and performance of dairy cattle during the pre- and post-partum periods under conditions of this study.

The plasma free amino acid dose-response technique: A proposed methodology for determining lysine relative bioavailability of rumen-protected lysine supplements.

Estimates of Lys bioavailability of rumen-protected Lys (RP-Lys) supplements are often obtained using in vitro or 2-step in situ techniques, with little to no data determining efficacy and bioavailability in vivo. The objective of this study was to further evaluate and refine the use of the plasma free AA dose-response technique as a method for determining Lys relative bioavailability of RP-Lys supplements. Thirteen dose-response Latin square studies using 87 lactating, ruminally cannulated multiparous Holstein cows (days in milk from 55 to 315 and milk yield from 12 to 62 kg/d at the start of the studies) were conducted to measure the relative bioavailability of RP-Lys supplements. Intestinal (1 study) and abomasal (12 studies) infusions of Lys ranged from 0 to 84 g/d, and experimental periods ranged from 4 to 21 d. Basal diets were formulated to be adequate in metabolizable Met, but varied in predicted metabolizable Lys (5.04 to 6.81% of metabolizable protein). One to 4 daily blood samples were taken from the coccygeal vessels for 1 to 3 consecutive days in each period. Plasma Lys concentration in cows assigned to the control treatment (0 g/d Lys) ranged from 1.83 to 5.21% of total plasma AA, whereas that from cows duodenally or abomasally infused with Lys ranged from 2.53 to 7.51% of total plasma AA. Results from studies involving more than 2 amounts of infused Lys confirmed linearity of response. The following variables were regressed against the plasma Lys dose-response slopes generated from the Lys infusion treatments to examine their effects on the magnitude of the slopes: plasma Lys concentration of the control diet, plasma Lys concentration at the greatest amount of infused Lys, net energy of lactation and metabolizable protein balances, metabolizable protein supply, days in milk, milk yield, milk concentrations of fat, true protein, and lactose, milk true protein yield, and dry matter intake. The variable having the greatest effect on the magnitude of the dose-response slope was the plasma Lys concentration at the greatest amount infused. The relative bioavailability of evaluated RP-Lys supplements using the plasma free AA dose-response technique ranged from 5 to 87%. It was concluded that plasma free Lys increases in a linear fashion to increasing amounts of absorbed Lys and that the dose-response technique is an appropriate technique for evaluating RP-Lys supplements.

Production performance and milk fatty acid profile in grazing dairy cows offered ground corn or liquid molasses as the sole supplemental nonstructural carbohydrate source.

The objective of this study was to compare the effects of ground corn or liquid molasses fed as the sole supplemental nonstructural carbohydrate (NSC) source on production performance, milk fatty acid (FA) profile, grazing behavior, and N metabolism in grazing dairy cows. A strip-grazing management system was used, with cows offered a new strip of fresh herbage after each milking, resulting in approximately 16 h of access to pasture daily. Animals were fed a diet formulated to yield an 86:14 forage-to-concentrate ratio consisting [dry matter (DM) basis] of 74% mixed grass-legume herbage, 12% mixed-mostly legume baleage, 12% NSC source, and 2% mineral-vitamin premix. Twenty Jersey cows averaging (mean ± standard deviation) 121 ± 73 d in milk in the beginning of the study were randomly assigned to 1 of 2 herbage supplementation treatments: (1) baleage plus ground corn (B+GC) or (2) baleage + liquid molasses (B+LM). Both NSC sources were fed at a flat rate of 1.6 kg of DM/cow daily. The study lasted from June to September for a total of 15 wk with data and sample collection conducted in wk 3, 7, 12, and 15. Milk samples for FA analysis were collected in wk 2, 4, 6, 8, 9, 11, and 13. Data were analyzed using the MIXED procedure of SAS (SAS Institute Inc., Cary, NC) for a randomized complete block design with repeated measures over time. Treatment × week interactions were observed for supplement DM intake, milk urea N, bite rate, urinary excretion of uric acid, and milk FA (e.g., 17:0, 18:0, cis-9,trans-11 18:2). Supplement DM intake was greatest in cows fed B+LM in wk 7, 12, and 15. Compared with cows fed B+GC, those fed B+LM had lower concentrations of milk urea N in wk 7 and 15. Milk yield, concentrations and yields of milk components, and plasma concentrations of essential AA, except Met, which was lowest with feeding B+LM, were not affected by supplementation. The plasma concentration of urea N was lowest with feeding B+LM. Cows fed B+GC spent more time grazing than those fed B+LM. Feeding B+GC increased cis-9 18:1 FA and most trans-18:1 FA in milk, whereas B+LM increased Σ odd-chain FA, Σ n-3 FA, and the trans-11 18:1 to trans-10 18:1 ratio, and decreased the n-6 to n-3 ratio. Based on current results, B+LM can entirely replace B+GC without negatively affecting milk yield or yields and concentrations of milk fat and true protein, while decreasing milk urea N, plasma urea N, and the milk trans-11 18:1 to trans-10 18:1 ratio, and increasing Σ n-3 FA.

Production and nitrogen utilization in lactating dairy cows fed ground field peas with or without ruminally protected lysine and methionine.

Previous research has shown that cows fed ≥24% of the diet dry matter (DM) as field peas decreased milk yield as well as concentration and yield of milk protein, possibly due to reduced DM intake and limited supply of Lys and Met. Twelve multiparous and 4 primiparous lactating Holstein cows were randomly assigned to 1 of 4 diets in a replicated 4 × 4 Latin square design. The diets contained (DM basis) 34.8% corn silage, 15.2% grass-legume silage, 5.9% roasted soybean, 2.4% mineral-vitamin premix, 2.0% alfalfa pellets, and either (1) 36% ground corn, 2.4% soybean meal, and 1.3% urea (UR), (2) 29.7% ground corn, 9.8% soybean meal, 0.13% ruminally protected (RP) Lys, and 0.07% RP-Met (CSBAA), (3) 25% ground field peas, 12.3% ground corn, and 2.4% soybean meal (FP), or (4) FP supplemented with 0.15% RP-Lys and 0.05% RP-Met (FPAA). Our objective was to test the effects of FP versus UR, FPAA versus CSBAA, and FPAA versus FP on milk yield and composition, N utilization, nutrient digestibility, ruminal fermentation profile, and plasma concentration of AA. Milk yield did not differ across diets. Compared with cows fed UR, those fed FP had greater DM intake, concentration and yield of milk true protein, apparent total-tract digestibility of fiber, urinary excretion of purine derivatives, and concentrations of total volatile fatty acids in the rumen and Lys in plasma, and less milk urea N and ruminal NH3-N. The concentration of milk urea N, as well as the concentration and yield of milk fat increased in cows fed FPAA versus CSBAA. Moreover, cows fed FPAA had greater ruminal concentration of total volatile fatty acids, increased proportions of acetate and isobutyrate, and decreased proportions of propionate and valerate than those fed CSBAA. The plasma concentrations of His, Leu, and Phe decreased, whereas plasma Met increased and plasma Lys tended to increase in cows fed FPAA versus CSBAA. Concentration of milk true protein, but not yield, was increased in cows fed FPAA versus FP. However, cows fed FPAA showed decreased concentrations of His and Leu in plasma compared with those fed FP. Overall, compared with the CSBAA diet, feeding FPAA did not negatively affect milk yield and milk protein synthesis. Furthermore, RP-Lys and RP-Met supplementation of the FP diet did not improve milk yield or milk protein synthesis, but decreased urinary urea N excretion.

Effect of feeding warm-season annuals with orchardgrass on ruminal fermentation and methane output in continuous culture.

A 4-unit, dual-flow continuous culture fermentor system was used to assess nutrient digestibility, volatile fatty acids (VFA) production, bacterial protein synthesis, and methane (CH4) output of warm-season annual grasses. Treatments were randomly assigned to fermentors in a 4 × 4 Latin square design using 7 d for adaptation to treatment and 3 d for sample collection. Treatments were (1) 100% orchardgrass (Dactylis glomerata L.; ORD); (2) 50% orchardgrass + 50% Japanese millet [Echinochloa esculenta (A. Braun) H. Scholz; MIL]; (3) 50% orchardgrass + 50% brown midrib sorghum × sudangrass (Sorghum bicolor L. Moench × S. bicolor var. sudanense; SSG]; or (4) 50% orchardgrass + 25% millet + 25% sorghum × sudangrass (MIX). Fermentors were fed 60 g of dry matter (DM)/d in equal portions of herbage 4 times daily (0730, 1030, 1400, and 1900 h). To replicate a typical 12-h pasture rotation, fermentors were fed the orchardgrass at 0730 and 1030 h and the individual treatment herbage (orchardgrass, Japanese millet, sorghum × sudangrass, or 50:50 Japanese millet and sorghum × sudangrass) at 1400 and 1900 h. Gas samples for CH4 analysis were collected 6 times daily at 0725, 0900, 1000, 1355, 1530, and 1630 h. Fermentor pH was determined at the time of feeding, and fermentor effluent samples for NH3-N and VFA analyses were taken daily at 1030 h on d 8, 9, and 10. Samples were also analyzed for DM, organic matter (OM), crude protein, and fiber fractions to determine nutrient digestibilities. Bacterial efficiency was estimated by dividing bacterial N by truly digested OM. True DM and OM digestibilities and pH were not different among treatments. Apparent OM digestibility was greater in ORD than in MIL and SSG. The concentration of propionate was greater in ORD than in SSG and MIX, and that of butyrate was greatest in ORD and MIL. Methane output was greatest in MIL, intermediate in ORD, and lowest in SSG and MIX. Nitrogen intake did not differ across treatments, whereas bacterial N efficiency per kilogram of truly digestible OM was greatest in MIL, intermediate in SSG and MIX, and lowest in ORD. True crude protein digestibility was greater in ORD versus MIL, and ORD had lower total N, non-NH3-N, bacterial N, and dietary N in effluent flows than MIL. Overall, we detected little difference in true nutrient digestibility; however, SSG and MIX provided the lowest acetate to propionate ratio and lower CH4 output than MIL and ORD. Thus, improved warm-season annual pastures (i.e., brown midrib sorghum × sudangrass) could provide a reasonable alternative to orchardgrass pastures during the summer months when such perennial cool-season grass species have greatly reduced productivity.

Effect of starchy or fibrous carbohydrate supplementation of orchardgrass on ruminal fermentation and methane output in continuous culture.

A 4-unit dual-flow continuous culture fermentor system was used to assess the effects of supplementing orchardgrass (Dactylis glomerata L.) with 2 levels [5 and 10% of total dry matter (DM) fed] of starchy (barley grain, BAR) or fibrous (beet pulp, BP) carbohydrate sources on nutrient digestibility, volatile fatty acid (VFA) production, bacterial protein synthesis, and CH4 output. Treatments were randomly assigned to fermentors in a 4×4 Latin square design with a 2×2 factorial arrangement using 7 d for microbial adaptation and 3 d for sample collection. Treatments included (1) 57g of DM orchardgrass + 3g of DM BAR, (2) 54g of DM orchardgrass + 6g of DM BAR, (3) 57g of DM orchardgrass + 3g of DM BP, or (4) 54g of DM orchardgrass + 6g of DM BP. Feedings occurred at 0900, 1030, 1400, and 1900h throughout four 10-d periods. Gas samples for CH4 analysis were collected 6 times daily at 0725, 0900, 1000, 1355, 1530, and 1630h. Fermentor samples for pH, NH3-N, and VFA analysis were taken on d 8, 9, and 10. Samples were also analyzed for DM, organic matter, crude protein, purines, neutral detergent fiber, and acid detergent fiber to determine nutrient digestibilities and estimation of bacterial protein synthesis. Apparent and true DM and organic matter digestibilities were not affected by supplement source. Apparent neutral and acid detergent fiber digestibilities were greater for BAR than BP. Conversely, apparent crude protein digestibility was greater for BP than BAR. Mean and maximum pH tended to be greatest for BAR than BP. Minimum pH was greater at the lower level (5% of diet DM) of supplementation. Barley produced greater concentrations of total VFA and acetate, whereas BP had greater daily outputs of CH4. Significant supplement type × level interactions were found for bacterial N flow and efficiency. Overall, supplementing orchardgrass with BP improved crude protein digestibility, reduced fiber digestibility and total VFA concentration, but increased CH4 output compared with BAR.

Incremental amounts of ground flaxseed decrease milk yield but increase n-3 fatty acids and conjugated linoleic acids in dairy cows fed high-forage diets(1).

The objective of this study was to investigate the effect of incremental amounts of ground flaxseed (GFX) on milk yield and concentrations and yields of milk components, milk fatty acids (FA) profile, ruminal metabolism, and nutrient digestibility in dairy cows fed high-forage diets. Twelve multiparous Jersey cows averaging (mean ± SD) 112±68d in milk and 441±21kg of body weight and 8 primiparous Jersey cows averaging 98±43d in milk and 401±43kg of body weight were randomly assigned to treatment sequences in a replicated 4×4 Latin square design. Each period lasted 21d with 14d for diet adaptation and 7d for data and sample collection. Treatments were fed as a total mixed ration (63:37 forage-to-concentrate ratio) with corn meal and soybean meal replaced by incremental levels (i.e., 0, 5, 10, or 15% diet dry matter) of GFX. The ruminal molar proportions of acetate and butyrate decreased linearly with GFX supplementation, whereas the ruminal molar proportion of propionate increased linearly resulting in decreased acetate-to-propionate ratio. Apparent total-tract digestibilities of nutrients either decreased (dry matter) or tended to decrease (organic matter, neutral detergent fiber, acid detergent fiber) linearly in cows fed GFX. Milk yield decreased linearly in cows fed increasing amounts of GFX, which is explained by the linear reduction in dry matter intake. Except for the concentrations of milk protein and urea N, which decreased linearly with GFX supplementation, no other changes in the concentration of milk components were observed. However, yields of milk protein and fat decreased linearly with GFX supplementation. The linear decrease in the yields of milk fat and protein are explained by reduced milk yield, whereas that in milk urea N is explained by decreased crude protein intake. No treatment effects were observed for plasma urea N and nonesterified fatty acids, serum cortisol, and body weight change. Milk odd- and branched-chain FA and saturated FA decreased linearly with GFX supplementation. Milk trans-11 18:1, α-linolenic acid, cis-9,trans-11 18:2, and the sum of n-3 FA all increased linearly and quadratically, whereas the milk ratio of n-6 to n-3 decreased linearly in cows fed GFX. Overall, compared with the control diet (0% GFX), the diet with 15% GFX supplementation resulted in the lowest milk yield but highest milk proportions and yields (data not shown) of cis-9,trans-11 18:2 and n-3 FA.

Antiulcerogenic Activity and Toxicity of Bauhinia holophylla Hydroalcoholic Extract.

Several species of Bauhinia are used in traditional medicine for the treatment of gastrointestinal diseases, diabetes, and inflammation, among other conditions. The aim of this study was to investigate the antiulcer effect of a hydroalcoholic extract from the leaves of B. holophylla. The chemical profile of the extract was determined by HPLC-PAD-ESI-IT-MS. A dose-effect relation was constructed using the ethanol-induced gastric ulcer model in male Wistar rats. Histological analyses and studies of antioxidant and anti-inflammatory activities were performed in stomach samples. The involvement of SH compounds, NO, K(+) ATP channels, and α 2-adrenergic receptors in the gastroprotective effect was evaluated. A toxicity study was performed with a single oral dose of 5000 mg/kg. The extract was composed mainly of cyanoglucoside and flavonol-O-glycosides derivatives of quercetin and myricetin. SH compounds, NO release, K(+) ATP channel activation, and presynaptic α 2-adrenergic receptor stimulation each proved to be involved in the antiulcer effect. The levels of GSH and activity of GR and GPx were increased, and the levels of TNF-α, IL-6 and IL-10 were modulated. There was an antidiarrheal effect and there were no signs of toxicity. B. holophylla presents antiulcer activity mainly by decreasing oxidative stress and attenuating the inflammatory response, without inducing side effects.