Proteomic analysis reveals greater abundance of complement and inflammatory proteins in subcutaneous adipose tissue from postpartum cows treated with sodium salicylate.

This study aimed to investigate sodium salicylate (SS) treatment effects on the proteome of adipose tissue (AT) in postpartum cows. Twenty Holstein cows were assigned to control (CON, n = 10) or SS (n = 10) provided via drinking water (2.3 g/L) during the first 7 d of lactation. Subcutaneous AT was collected on d 7 of treatment and label-free quantitative shotgun proteomics and immunoblotting were analyzed in a subset of 5 AT per group. Eighty out of 1422 proteins (5.6%) were differentially abundant between CON and SS [fold change ±1.5, P < 0.05]. Top canonical pathways differing between CON and SS (Ingenuity) were complement system, interleukin-10 signaling, and acute phase response signaling. The abundances of complement C1r, C1qC, C1qB and C6 were greater in SS than CON. Regarding IL-10 signaling, the abundances of BLVRB, STAT3, and lipopolysaccharide binding protein (LBP) were greater in SS AT compared to CON. Immunoblots revealed increased abundance of paraoxanase-1 and tumor necrosis factor-alpha, as well as a tendency for greater abundance of cluster differentiation 172a in SS AT, which may indicate of increased macrophage infiltration. SS treatment postpartum likely promotes inflammatory signaling in AT of dairy cows, perhaps due to immune cell recruitment. SIGNIFICANCE: This work demonstrates that treating early lactating cows with sodium salicylate, an anti-inflammatory agent that has been shown to have metabolic effects and increase milk production in dairy cows, affects the proteome of subcutaneous adipose tissue in early lactating dairy cows. Unexpectedly, sodium salicylate treatment enriched inflammatory pathways of the complement system, cytokine signaling, and acute phase response, as revealed by proteomic analysis of subcutaneous adipose tissues from cows at 7 d postpartum. These findings imply that SS treatment during the first 7 d of lactation likely promotes inflammatory signaling in AT of the dairy cow, perhaps due to immune cell recruitment. Tissue-specific impacts of systemic sodium salicylate requires further scrutiny.

Effects of chitosan and whole raw soybeans on ruminal fermentation and bacterial populations, and milk fatty acid profile in dairy cows.

The objective of this study was to evaluate whether providing chitosan (CHI) to cows fed diets supplemented with whole raw soybeans (WRS) would affect the nutrient intake and digestibility, ruminal fermentation and bacterial populations, microbial protein synthesis, N utilization, blood metabolites, and milk yield and composition of dairy cows. Twenty-four multiparous Holstein cows (141 ± 37.1 d in milk, 38.8 ± 6.42 kg/d of milk yield; mean ± SD) were enrolled to a 4 × 4 Latin square design experiment with 23-d periods. Cows were blocked within Latin squares according to milk yield, days in milk, body weight, and rumen cannula (n = 8). A 2 × 2 factorial treatment arrangement was randomly assigned to cows within blocks. Treatments were composed of diets with 2 inclusion rates of WRS (0 or 14% diet dry matter) and 2 doses of CHI (0 or 4 g/kg of dry matter, Polymar Ciência e Nutrição, Fortaleza, Brazil). In general, CHI+WRS negatively affected nutrient intake and digestibility of cows, decreasing milk yield and solids production. The CHI increased ruminal pH and decreased acetate to propionate ratio, and WRS reduced NH3-N concentration and acetate to propionate in the rumen. The CHI reduced the relative bacterial population of Butyrivibrio group, whereas WRS decreased the relative bacterial population of Butyrivibrio group, and Fibrobacter succinogenes, and increased the relative bacterial population of Streptococcus bovis. No interaction effects between CHI and WRS were observed on ruminal fermentation and bacterial populations. The CHI+WRS decreased N intake, microbial N synthesis, and N secreted in milk of cows. The WRS increased N excreted in feces and consequently decreased the N excreted in urine. The CHI had no effects on blood metabolites, but WRS decreased blood concentrations of glucose and increased blood cholesterol concentration. The CHI and WRS improved efficiency of milk yield of cows in terms of fat-corrected milk, energy-corrected milk, and net energy of lactation. The CHI increased milk concentration [g/100 g of fatty acids (FA)] of 18:1 trans-11, 18:2 cis-9,cis-12, 18:3 cis-9,cis-12,cis-15, 18:1 cis-9,trans-11, total monounsaturated FA, and total polyunsaturated FA. The WRS increased total monounsaturated FA, polyunsaturated FA, and 18:0 to unsaturated FA ratio in milk of cows. Evidence indicates that supplementing diets with unsaturated fat sources along with CHI negatively affects nutrient intake and digestibility of cows, resulting in less milk production. Diet supplementation with CHI or WRS can improve feed efficiency and increases unsaturated FA concentration in milk of dairy cows.

Ruminal biohydrogenation and abomasal flow of fatty acids in lactating cows fed diets supplemented with soybean oil, whole soybeans, or calcium salts of fatty acids.

Ruminants have a unique metabolism and digestion of unsaturated fatty acids (UFA). Unlike monogastric animals, the fatty acid (FA) profile ingested by ruminants is not the same as that reaching the small intestine. The objective of this study was to evaluate whole raw soybeans (WS) in diets as a replacer for calcium salts of fatty acids (CSFA) in terms of UFA profile in the abomasal digesta of early- to mid-lactation cows. Eight Holstein cows (80 ± 20 d in milk, 22.9 ± 0.69 kg/d of milk yield, and 580 ± 20 kg of body weight; mean ± standard deviation) with ruminal and abomasal cannulas were used in a 4 × 4 Latin square experiment with 22-d periods. The experiment evaluated different fat sources rich in linoleic acid on ruminal kinetics, ruminal fermentation, FA abomasal flow, and milk FA profile of cows assigned to treatment sequences containing a control (CON), with no fat source; soybean oil, added at 2.68% of diet dry matter (DM); WS, addition of WS at 14.3% of diet DM; and CSFA, addition of CSFA at 2.68% of diet DM. Dietary fat supplementation had no effect on nutrient intake and digestibility, with the exception of ether extract. Cows fed fat sources tended to have lower milk fat concentration than those fed CON. In general, diets containing fat sources tended to decrease ruminal neutral detergent fiber digestibility in relation to CON. Cows fed WS had lower ruminal digestibility of DM and higher abomasal flow of DM in comparison to cows fed CSFA. As expected, diets containing fat supplements increased FA abomasal flow of C18:0 and total FA. Cows fed WS tended to present a higher concentration of UFA in milk when compared with those fed CSFA. This study suggests that under some circumstances, abomasal flow of UFA in early lactation cows can be increased by supplementing their diet with fat supplements rich in linoleic acid, regardless of rumen protection, with small effects on ruminal DM digestibility.

Soybean whole-plant ensiled with chitosan and lactic acid bacteria: Microorganism counts, fermentative profile, and total losses.

Chitosan is a biopolymer derived from chitin deacetylation, present in the exoskeleton of crustaceans and insects. Chitosan has been evaluated as rumen modulator and silage additive due to its antimicrobial properties. The objective of this study was to determine the effects of both chitosan and a bacterial additive on microbiological quality, chemical composition, nutrient in vitro degradation, fermentative profile, and total losses of whole-soybean plant silage (SS) harvested at R6 stage. Four treatments in a factorial arrangement were randomly assigned to 40 experimental minisilos as no additives (CON), 8 g/t fresh forage of microbial inoculant (INO; Kera SIL, Kera Nutrição Animal, Bento Gonçalves, Brazil); 5 g/kg of fresh forage chitosan (CHI); and CHI + INO. Microbial inoculant was composed of Lactobacillus plantarum (4.0 × 1010 cfu/g) and Propionibacterium acidipropionici (2.6 × 1010 cfu/g). The CHI and INO alone increased counts of lactic bacteria and anaerobic bacteria and decreased counts of mold and yeast in SS. The CHI or INO alone increased in vitro degradation of dry matter, crude protein, and neutral detergent fiber, and decreased nonfiber carbohydrate content of SS. Chitosan increased NH3-N and lactate concentrations and decreased ethanol concentration in SS. The CHI increased dry matter recovery from SS; INO increased silage aerobic stability. The combination of CHI+INO showed the lowest value of gas losses. In general, the combination of CHI and INO had small positive effects on gas losses of SS; however, both CHI or INO alone improved nutrient in vitro degradation and decreased mold and yeast in SS. Chitosan or INO utilization improves SS quality.

Different durations of whole raw soybean supplementation during the prepartum period: Measures of cellular immune function in transition cows.

The objective of this study was to evaluate different durations of whole raw soybean (WS) supplementation (diet rich in n-6 fatty acid) during the prepartum period on cellular immune function of dairy cows in the transition period and early lactation. Thirty-one Holstein cows were used in a completely randomized design and assigned to 4 experimental groups (G) [G90, G60, G30, and G0 (control)] supplemented with a diet containing 12% of WS from 90, 60, 30 and 0 d relative to the calving date, respectively. Cows were dried off 60 d before the expected calving date. After parturition, all cows were fed a diet containing 12% of WS until 84 DIM. Blood samples were collected before the morning feeding (d -56 ± 2, -28 ± 2, -14 ± 2, -7 ± 2, at the day of partum, 7 ± 2, 14 ± 2, 28 ± 2, and 56 ± 2 relative to parturition). Cell phenotyping and phagocytosis assays were carried out using monoclonal antibodies and flow cytometry technique. Duration of WS supplementation linearly increased the percentage of blood CD3+ cells, as well as increased the percentage of blood CD8+ cells in the postpartum period, notably in G30, whereas the lowest values were observed in G0. Further, the duration of WS supplementation linearly increased the reactive oxygen species median fluorescence intensity of CH138+ cells after phagocytizing Staphylococcus aureus in the postpartum period. Longer periods of WS supplementation linearly increased phagocytosis median fluorescence intensity of CH138+ cells in the prepartum period of cows. Duration of WS supplementation linearly increased the percentage of blood CD14+ cells producing reactive oxygen species when stimulated either by Staph. aureus or Escherichia coli in the postpartum period. In conclusion, longer periods of WS supplementation during late lactation and the dry period (beginning on d 90 of the expected calving date) alter the leukocyte population and improve neutrophil immune response in the postpartum period with no detrimental effects on cow performance.

Different durations of whole raw soybean supplementation during the prepartum period: Milk fatty acid profile and oocyte and embryo quality of early-lactating Holstein cows.

The objective of this study was to evaluate different durations of whole raw soybeans (WS) supplementation during the prepartum period on nutrient digestibility, milk yield and composition, energy balance, blood metabolites, and oocyte and embryo quality of transition cows. Thirty-one Holstein cows were used in a completely randomized design and assigned to 4 experimental groups (G): G90, G60, G30, and G0 (control), supplemented with a diet containing 12% of WS from 90, 60, 30, and 0 d relative to the calving date, respectively. Cows were dried off 60 d before the expected calving date. After parturition, all cows were fed a diet containing 12% of WS until 84 DIM. Blood samples were collected on d -49, -35, -21, -14, -7, 0, 7, 14, 21, 35, and 70 relative to partum. Ovum pick-ups were performed on d 21 ± 3, 42 ± 7, 63 ± 7, and 84 ± 7 of lactation. Different durations of WS supplementation did not affect DMI and apparent total-tract digestibility in either the pre- or postpartum periods. Duration of WS supplementation had no effect on milk yield and composition nor energy balance of cows. However, the duration of WS supplementation had several effects on milk fatty acid (FA) profile of cows, including a linear decrease in concentrations of cis-9 C18:1, unsaturated C18, total monounsaturated, and unsaturated FA. Further, the milk contents of cis-9,cis-12 C18:2 FA, cis-9,trans-11 C18:2 FA, and total polyunsaturated FA were increased when WS were fed to cows from 30 d but not from 60 or 90 d of the expected calving date. The length of WS supplementation in the prepartum period linearly increased blood cholesterol concentration of cows during the prepartum period, but it had no effect on blood glucose and nonesterified FA concentrations in the pre- and postpartum periods. Duration of WS supplementation during the prepartum period increased the average number of grade 2 oocytes, notably in G60, but it had no effect on embryo production and cleavage proportion of early-lactation cows. The duration of WS supplementation in the prepartum period had no effect on milk yield and energy balance of the subsequent lactation, but it altered milk FA profile in early lactation by decreasing unsaturated FA content, notably when starting to supplement WS at 90 and 60 d from the expected calving date. Our results also showed that the duration of WS supplementation during the prepartum period does not improve oocyte quality in the subsequent lactation of cows.

Influence of polymer-coated slow-release urea on total tract apparent digestibility, ruminal fermentation and performance of Nellore steers.

OBJECTIVE: Two experiments were performed to evaluate the effects of coated slow-release urea on nutrient digestion, ruminal fermentation, nitrogen utilization, blood glucose and urea concentration (Exp 1), and average daily gain (ADG; Exp 2) of steers. METHODS: Exp 1: Eight ruminally fistulated steers [503±28.5 kg body weight (BW)] were distributed into a d 4×4 Latin square design and assigned to treatments: control (CON), feed grade urea (U2), polymer-coated slow-release urea A (SRA2), and polymer-coated slow-release urea B (SRB2). Dietary urea sources were set at 20 g/kg DM. Exp 2: 84 steers (350.5±26.5 kg initial BW) were distributed to treatments: CON, FGU at 10 or 20 g/kg diet DM (U1 and U2, respectively), coated SRA2 at 10 or 20 g/kg diet DM (SRA1 and SRA2, respectively), and coated SRB at 10 or 20 g/kg diet DM (SRB1 and SRB2, respectively). RESULTS: Exp 1: Urea treatments (U2+SRA2+SRB2) decreased (7.4%, p = 0.03) the DM intake and increased (11.4%, p<0.01) crude protein digestibility. Coated slow-release urea (SRA2+SRB2) showed similar nutrient digestibility compwared to feed grade urea (FGU). However, steers fed SRB2 had higher (p = 0.02) DM digestibility compared to those fed SRA2. Urea sources did not affect ruminal fermentation when compared to CON. Although, coated slow-release urea showed lower (p = 0.01) concentration of NH3-N (-10.4%) and acetate to propionate ratio than U2. Coated slow-release urea showed lower (p = 0.02) urinary N and blood urea concentration compared to FGU. Exp 2: Urea sources decreased (p = 0.01) the ADG in relation to CON. Animals fed urea sources at 10 g/kg DM showed higher (12.33%, p = 0.01) ADG compared to those fed urea at 20 g/kg DM. CONCLUSION: Feeding urea decreased the nutrient intake without largely affected the nutrient digestibility. In addition, polymer-coated slow-release urea sources decreased ruminal ammonia concentration and increased ruminal propionate production. Urea at 20 g/kg DM, regardless of source, decreased ADG compared both to CON and diets with urea at 10 g/kg DM.

Short communication: Partial replacement of ground corn with algae meal in a dairy cow diet: Milk yield and composition, nutrient digestibility, and metabolic profile.

This study was undertaken to evaluate the effects of partially replacing dietary ground corn with a microalgae meal from Prototheca moriformis (composed of deoiled microalgae and soyhulls) on milk yield and composition, nutrient intake, total-tract apparent digestibility, and blood profile of lactating dairy cows. Twenty multiparous Holstein cows (57.7±49.4d in milk, 25.3±5.3 of milk yield, and 590±71kg of live weight at the start of experiment, mean ± standard deviation) were used in a cross-over design experiment, with 21-d periods. Diets were no microalgae meal (CON) or 91.8g/kg of microalgae meal partially replacing dietary ground corn (ALG). Cows showed similar milk yield and composition. The 3.5% fat-corrected milk production was 30.2±1.34kg/d for CON and 31.1±1.42kg/d for ALG. Despite cows having similar dry matter intake, ALG increased neutral detergent fiber and ether extract intake. In addition, cows fed ALG exhibited higher ether extract digestibility. No differences were detected in glucose, urea, amino-aspartate transferase, and gamma-glutamyl transferase blood concentrations. Feeding ALG increased the total cholesterol and high-density lipoprotein in blood compared with CON. The microalgae meal may partially replace ground corn in diets of lactating cows without impairing the animal's performance.

Effects of whole flaxseed, raw soybeans, and calcium salts of fatty acids on measures of cellular immune function of transition dairy cows.

The objective of the current study was to evaluate the effects of supplemental n-3 and n-6 fatty acid (FA) sources on cellular immune function of transition dairy cows. Animals were randomly assigned to receive 1 of 4 diets: control (n=11); whole flaxseed (n-3 FA source; n=11), 60 and 80g/kg of whole flaxseed [diet dry matter (DM) basis] during pre- and postpartum, respectively; whole raw soybeans (n-6 FA source; n=10), 120 and 160g/kg of whole raw soybeans (diet DM basis) during pre- and postpartum, respectively; and calcium salts of unsaturated FA (Megalac-E, n-6 FA source; n=10), 24 and 32g/kg of calcium salts of unsaturated FA (diet DM basis) during pre- and postpartum, respectively. Supplemental FA did not alter DM intake and milk yield but increased energy balance during the postpartum period. Diets containing n-3 and n-6 FA sources increased phagocytosis capacity of leukocytes and monocytes and phagocytosis activity of monocytes. Furthermore, n-3 FA source increased phagocytic capacity of leukocytes and neutrophils and increased phagocytic activity in monocytes and neutrophils when compared with n-6 FA sources. Supplemental FA effects on adaptive immune system included increased percentage of T-helper cells, T-cytotoxic cells, cells that expressed IL-2 receptors, and CD62 adhesion molecules. The results of this study suggest that unsaturated FA can modulate innate and adaptive cellular immunity and trigger a proinflammatory response. The n-3 FA seems to have a greater effect on phagocytic capacity and activity of leukocytes when compared with n-6 FA.

Effects of flaxseed, raw soybeans and calcium salts of fatty acids on apparent total tract digestibility, energy balance and milk fatty acid profile of transition cows.

Oilseeds offer some protection to the access of ruminal microorganisms and may be an alternative to calcium salts of fatty acids (FA), which are not fully inert in the ruminal environment. This study aimed to evaluate the effects of different sources of FA supplementation on apparent total tract nutrient digestibility, milk yield and composition, and energy balance (EB) of cows during the transition period and early lactation. We compared diets rich in C18:2 and C18:3 FA. Multiparous Holstein cows were randomly assigned to receive one of the four diets: control (n=11); whole flaxseed (WF, n=10), 60 and 80 g/kg (diet dry matter (DM) basis) of WF during the prepartum and postpartum periods, respectively; whole raw soybeans (WS, n=10), 120 and 160 g/kg (diet DM basis) of WS during the prepartum and postpartum periods, respectively; and calcium salts of unsaturated fatty acids (CSFA, n=11), 24 and 32 g/kg (diet DM basis) of CSFA during the prepartum and postpartum periods, respectively. Dry cows fed WF had higher DM and net energy of lactation (NEL) intake than those fed WS or CSFA. The FA supplementation did not alter DM and NDF apparent total tract digestibility, dry cows fed WF exhibited greater NDF total tract digestion than cows fed WS or CSFA. Feeding WS instead of CSFA did not alter NEL intake and total tract digestion of nutrients, but increased milk fat yield and concentration. Calculated efficiency of milk yield was not altered by diets. FA supplementation increased EB during the postpartum period. Experimental diets increased long-chain FA (saturated and unsaturated FA) in milk. In addition, cows fed WS and CSFA had higher C18:1 trans-11 FA and C18:2 cis, and lower C18:3 FA in milk than those fed WF. Furthermore, cows fed CSFA had higher C18:1 trans-11 and cis-9, trans-11 FA than cows fed WS. Although supplemental C18:2 and C18:3 FA did not influence the milk yield of cows, they positively affected EB and increased unsaturated long-chain FA in milk fat.

Effect of raw soya bean particle size on productive performance and digestion of dairy cows.

Differing soya bean particle sizes may affect productive performance and ruminal fermentation due to the level of fatty acid (FA) exposure of the cotyledon in soya bean grain and because the protein in small particles is more rapidly degraded than the protein in large particles, which influence ruminal fibre digestion and the amounts of ruminally undegradable nutrients. The objective of this experiment was to investigate the effects of raw soya bean particle size on productive performance, digestion and milk FA profile of dairy cows. Twelve Holstein cows were assigned to three 4 × 4 Latin squares with 21-day periods. At the start of the experiment, cows were 121 days in milk (DIM) and yielded 30.2 kg/day of milk. Cows were fed 4 diets: (i) control diet (CO), without raw soya bean; (ii) whole raw soya bean (WRS); (iii) cracked raw soya bean in Wiley mill 4-mm screen (CS4); and (iv) cracked raw soya bean in Wiley mill 2-mm screen (CS2). The inclusion of soya beans (whole or cracked) was 200 g/kg on dry matter (DM) basis and partially replaced ground corn and soya bean meal. Uncorrected milk yield and composition were not influenced by experimental diets; however, fat-corrected milk (FCM) decreased when cows were fed soya bean treatments. Soya bean diets increased the intake of ether extract (EE) and net energy of lactation (NEL ), and decreased the intake of DM and non-fibre carbohydrate (NFC). Ruminal propionate concentration was lower in cows fed WRS than cows fed CS2 or CS4. Cows fed cracked raw soya bean presented lower nitrogen in faeces than cows fed WRS. The milk of cows fed WRS, CS2 and CS4 presented higher unsaturated FA than cows fed CO. The addition of raw soya bean in cow diets, regardless of the particle size, did not impair uncorrected milk yield and nutrient digestion, and increased the concentration of unsaturated FA in milk. Cows fed cracked raw soya bean presented similar productive performance to cows fed whole raw soya bean.

Total tract nutrient digestion and milk fatty acid profile of dairy cows fed diets containing different levels of whole raw soya beans.

Whole oilseeds such as soya beans have been utilized in dairy rations to supply additional fat and protein. However, antinutritional components contained in soya beans, such as trypsin inhibitors and haemagglutinins (lectins) may alter digestibility of nutrients and consequently affect animal performance. The objective of the present experiment was to quantify the effect of different levels of whole raw soya beans in diets of dairy cows on nutrient intake, total tract digestion, nutrient balances and milk yield and composition. Sixteen mid to late-lactation cows (228 ± 20 days in milk; mean ± SD) were used in four replicated 4 × 4 Latin square design experiment with 21-d periods. Cows were assigned to each square according to milk yield and DIM. The animals were randomly allocated to treatments: control (without soya beans addition; CO), WS9, WS18 and WS27, with addition of 9%, 18% and 27% of whole raw soya bean in diet on a dry matter (DM) basis respectively. All diets contained identical forage and concentrate components and consisted of maize silage and concentrate based on ground corn and soya beans at a ratio of 60:40. There were no differences in OM, CP, NDF and NEL intakes (kg/day and MJ/day) among the treatments (p > 0.05). However, DM and NFC intakes were negatively affected (p = 0.04 and p < 0.01, respectively) and ether extract (EE) intake was positively affected (p < 0.01). Total tract digestion increased linearly with whole raw soya beans for EE (p < 0.01) and NDF (p = 0.01). The excretion (kg/day) of digested soya beans grains increased linearly according to addition of whole raw soya beans. However, the nutritive characteristics of excreted grains were not altered. Milk (kg), milk lactose (kg) and protein (kg) yield decreased linearly (p < 0.01, p < 0.01 and p = 0.04, respectively) milk fat content (%) increased linearly (p < 0.01) with whole raw soya beans inclusion. Increasing addition of whole raw soya beans affected milk fatty acid profile with a linear decrease of cis-9-trans 11CLA and total saturated FA; and linear increase of total unsaturated and C18:3 FA. Energy balance was positively affected (p = 0.03) by whole raw soya beans as well as efficiency of NEL milk/DE intake (p = 0.02). Nitrogen balance and microbial protein synthesis were not affected by whole raw soya beans. Increasing doses of whole raw soya beans decreased dry matter intake and milk yield, however, led to an increase of unsaturated acids in milk and higher milk fat concentration.