Plasma and milk metabolomics revealed changes in amino acid metabolism in Holstein dairy cows under heat stress.

Our understanding of metabolic alterations triggered by heat stress is incomplete, which limits the designing of nutritional strategies to mitigate negative productive and health effects. Thus, this study aimed to explore the metabolic responses of heat-stressed dairy cows to dietary supplementation with vitamin D3/Ca and vitamin E/Se. Twelve multiparous Holstein cows were enrolled in a split-plot Latin square design with two distinct vitamin E/Se supplementation levels, either at a low (ESe-, n = 6, 11.1 IU/kg vitamin E and 0.55 mg/kg Se) or a high dose (ESe+, n = 6 223 IU/kg vitamin E and 1.8 mg/kg Se) as the main plot. Treatment subplots, arranged in a replicated 3 × 3 Latin square design, comprised heat challenge (Temperature Humidity Index, THI: 72.0-82.0) supplemented with different levels of vitamin D3/Ca: either low (HS/DCa-, 1 012 IU/kg and 0.73%, respectively) or high (HS/DCa+, 3 764 IU/kg and 0.97%, respectively), and a pair-fed control group in thermoneutrality (THI = 61.0-64.0) receiving the low dose of vitamin D3/Ca (TN). The liquid chromatography-mass spectrometry-based metabolome profile was determined in blood plasma and milk sampled at the beginning (day 0) and end (day 14) of each experimental period. The results were analyzed for the effect of (1) TN vs. HS/ESe-/DCa-, and (2) the vitamin E/Se and vitamin D3/Ca supplementation. No group or group × day effects were detected in the plasma metabolome (false discovery rate, FDR > 0.05), except for triglyceride 52:2 being higher (FDR = 0.03) on day 0 than 14. Taurine, creatinine and butyryl-carnitine showed group × day interactions in the milk metabolome (FDR ≤ 0.05) as creatinine (+22%) and butyryl-carnitine (+190%) were increased (P < 0.01) on day 14, and taurine was decreased (-65%, P < 0.01) on day 14 in the heat stress (HS) cows, compared with day 0. Most compounds were unaffected by vitamin E/Se or vitamin D3/Ca supplementation level or their interaction (FDR > 0.05) in plasma and milk, except for milk alanine which was lower (-69%, FDR = 0.03) in the E/Se+ groups, compared with E/Se-. Our results indicated that HS triggered more prominent changes in the milk than in the plasma metabolome, with consistent results in milk suggesting increased muscle catabolism, as reflected by increased creatinine, alanine and citrulline levels. Supplementing with high levels of vitamin E/Se or vitamin D3/Ca or their combination did not appear to affect the metabolic remodeling triggered by HS.

Endometrial gene expression in primiparous dairy cows at the end of the voluntary waiting period is affected by nutrition: Total mixed ration vs increasing levels of herbage allowance.

The study postulated that differential nutritional management during the early lactation period would be reflected in endometrial expression of genes related to embryo growth at the end of the voluntary waiting period. Thus, the effect of the combined use of total mixed ration (TMR) and grazing under different herbage allowances during the first 75 days post-partum (DPP) on endometrial gene expression was evaluated in primiparous dairy cows. Cows were blocked by body weight, age and body condition score and randomly assigned to three grazing treatments: high (HA, 30 kg DM per cow per day), medium (MA, 15 kg DM per cow per day) and low (LA, 7.5 kg DM per cow per day) herbage allowance (mixed pasture, 2,600 kg DM per ha) plus 8 kg DM of supplement or TMR (55% forage, 45% concentrate) fed ad libitum (TMR) from calving to 75 DPP. At 57 DPP, cows were synchronized for oestrus (day 0, 68 DPP) and at day 7, endometrial biopsies were obtained. The nutritional treatment did not affect insulin, IGF-1 and leptin concentrations on days 0, 4 or 7. Expression of IGF1, IGFBP3, IGFBP4, ADIPOR1 and ADIPOR2 mRNA was significantly affected by the nutritional treatment. Endometrial IGF1 and IGFBP4 mRNA were twofold greater in TMR and HA than MA and LA cows. Expression of IGFBP3 and ADIPOR1 mRNAs was greater in TMR and HA than MA cows, but did not differ from LA cows. All groups had greater expression of ADIPOR2 mRNA than MA cows. This study provided solid evidence of the importance of nutritional management during early lactation on uterine environment at the end of the voluntary waiting period. The greater expression of genes related to embryo growth and uterine function (IGF system, progesterone and adiponectin receptors) in cows fed diets maximizing energy intake suggests a favourable environment for embryonic growth, which may explain the improved reproductive performance of cows in good energy balance.

Effect of sorghum grain supplementation on glucose metabolism in cattle and sheep fed temperate pasture.

The aim of this work was to evaluate the effect of sorghum grain supplementation on plasma glucose, insulin and glucagon concentrations, and hepatic mRNA concentrations of insulin receptor (INSR), pyruvate carboxylase (PC), and phosphoenolpyruvate carboxykinase (PCK1) mRNA and their association with nutrient intake, digestion and rumen volatile fatty acids (VFA) in cattle and sheep fed a fresh temperate pasture. Twelve Hereford × Aberdeen Angus heifers and 12 Corriedale × Milchschaf wethers in positive energy balance were assigned within each species to one of two treatments (n = 6 per treatment within specie): non-supplemented or supplemented with sorghum grain at 15 g/kg of their body weight (BW). Supplemented cattle had greater plasma glucose concentrations, decreased plasma glucagon concentrations and tended to have greater plasma insulin and insulin-to-glucagon ratio than non-supplemented ones. Hepatic expression of INSR and PC mRNA did not differ between treatments but PCK1 mRNA was less in supplemented than non-supplemented cattle. Supplemented sheep tended to have greater plasma glucagon concentrations than non-supplemented ones. Plasma glucose, insulin, insulin-to-glucagon ratio, and hepatic expression of INSR and PC mRNA did not differ between treatments, but PCK1 mRNA was less in supplemented than non-supplemented sheep. The inclusion of sorghum grain in the diet decreased PCK1 mRNA but did not affect PC mRNA in both species; these effects were associated with changes in glucose and endocrine profiles in cattle but not in sheep. Results would suggest that sorghum grain supplementation of animals in positive energy balance (cattle and sheep) fed a fresh temperate pasture would modify hepatic metabolism to prioritize the use of propionate as a gluconeogenic precursor.

Effects of herbage allowance of native grasslands in purebred and crossbred beef cows: metabolic, endocrine and hepatic gene expression profiles through the gestation-lactation cycle.

Our objective was to evaluate the metabolic, endocrine and hepatic mRNA profiles through the gestation-lactation cycle in purebred (PU: Angus and Hereford) and crossbred (CR: reciprocal F1 crosses) mutliparous beef cows (n=32), grazing on two herbage allowances of native pastures (2.5 v. 4 kg dry matter/kg BW; LO v. HI) and their associations with cow's productive performance (calf birth weight, milk production and commencement of luteal activity). Cow BW, body condition score (BCS) and blood samples were collected monthly, starting at -165 days relative to calving (days), and every 2 weeks after calving until +60 days of lactation. Liver biopsies were collected at -165, -75, -45, -15±10, and +15 and +60±3 days. Metabolic, endocrine and hepatic gene expression profiles, and calf birth weight, milk yield and postpartum commencement of luteal activity were evaluated. Overall, the most pronounced changes in metabolic, endocrine and hepatic gene expression occurred during winter gestation (-165 to -45 days), when all cows experienced the onset of a negative energy balance (decreased BCS, glucose and insulin, and increased non-esterified fatty acid concentrations, P<0.008). Concentrations of insulin and IGF-I were greater (P<0.037) in HI than in LO cows. However, serum IGF-I concentrations and hepatic growth hormone receptor (GHR) and IGF1 mRNA decreased (P<0.05) during the winter gestation period only in HI cows. Although IGF-I concentrations decreased (P<0.05) during the early postpartum (-15 v.+15 days) for all cows, the typical molecular mechanism that control the uncoupling of the growth hormone-IGF1 axis during the transition period of the dairy cattle (reduced hepatic GHR1A and IGF-I mRNA) was not observed in this study. The hepatic mRNA expression of key transcripts involved in gluconeogenesis and fatty-acid oxidation were upregulated (P<0.05) during winter gestation (from -165 to -45, -15 or +15 days, depending on the cow groups). Particularly, acyl-CoA oxidase-1 mRNA was greater for CR than for PU cows during winter gestation (-75 and -45 days), and fibroblast growth factor-21 mRNA was downregulated (P<0.01) only for HI cows during the transition (-15 v. 15 days) and lactation period (+15 to +60 days, P<0.01). These results, together with the greater BCS, estimated energy intake, increased milk yield and shorter commencement of luteal activity in HI than in LO, and in CR than in PU cows (P<0.018), would indicate that HI and CR cows were able to adapt more efficiently to changes in nutrient and energy supply through the gestation-lactation cycle.

Calf foetal and early life nutrition on grazing conditions: metabolic and endocrine profiles and body composition during the growing phase.

The aim of this study was to determine the effect of nutrition during foetal and lactation periods on calf growth and body composition, and their association with changes in metabolic and endocrine profiles during the calf first year of life on purebred (Hereford and Angus) and cross-bred (F1) dam offspring. Forty cross-bred calves and their dams (purebred--PU: Hereford and Angus, and cross-bred--CR: F1) were used in a randomized block design with a factorial arrangement of herbage allowance of native pastures (high: Hi-HA and low: Lo-HA), 4 vs. 2.5 kg dry matter/kg body weight (BW) and dam genotype (PU vs. CR). Calf BW and blood samples were collected monthly from birth to 380±15 days of age, and body composition was estimated by the urea dilution technique at weaning (142±15 days) and 380 days. Calf birthweight did not differ among groups but from birth to 380 days, and BW was reduced (p=0.046) in Lo-PU offspring. Although Lo-CR calves achieved similar BW than Hi-PU and Hi-CR offspring, they showed an increased fat in detriment of lean tissue deposition. At birth, plasma total protein was less (p=0.04), while plasma glucose, insulin or IGF-I tended or were greater (p<0.072) in Hi-HA than Lo-HA calves. Greater (p<0.03) plasma total protein and/or glucose concentrations during the first months of lactation were observed in CR offspring associated with the greater dam milk production. Although glucose concentrations did not differ among calf groups after weaning, plasma insulin was greater (p=0.004) in Hi-PU than other groups at 380 days. Consistent with the reduced BW, Lo-PU offspring presented the lowest (p=0.026) plasma IGF-I from birth to 380 days. Herbage allowance of native grasslands during calf foetal and lactation periods interacted with maternal heterosis to affect, in the short and/or long term, calf BW or body composition, and metabolic and endocrine profiles.

Effects of a short-term increase in the nutritional plane before the mating period on metabolic and endocrine parameters, hepatic gene expression and reproduction in primiparous beef cows on grazing conditions.

Sixty-four spring-calved primiparous crossbred cows paired by calving date and body condition score (BCS) at calving were used to study the effect of a short-term increase in the nutritional plane before the mating period on cow and calf performance, changes in metabolic and endocrine parameters and hepatic gene expression. At 48 ± 10 days post-partum (onset of nutritional treatment = day 0), cows were assigned to two treatments during 23 days: control (grazing of native pastures; NP; n = 31) and increased nutritional plane (NP improved with Lotus subbiflorous cv Rincon; IP; n = 33). Cow body weight (BW), BCS and total protein and albumin concentrations increased while urea and non-esterified fatty acids levels decreased from the beginning of the nutritional treatment in both groups, indicating the animal positive energy balance as forage growth and availability of pastures increased during spring. In addition, cow BW and BCS, as well as calf average daily gain and BW, were greater in IP than in NP cows groups. Insulin concentrations were less in IP than in NP (1.37 vs. 2.25 ± 0.26 µU/ml) because insulin increased owing to nutritional treatment only in NP cows. Hepatic insulin receptor mRNA at day 23 tended to be 1.5-fold greater, while insulin growth factor binding protein-3 mRNA expression was 1.7-fold greater in NP than in IP cows. Reproductive responses were not affected by nutritional treatment, but days to initiation of ovarian ciclicity (108 ± 10 days) were positively correlated with insulin concentrations. Grazing of improved NP for 23 days before the mating period did not improve cow reproductive performance but modified metabolic, endocrine and gene expression parameters, in agreement with greater nutrient and energy partitioning towards milk production, reflected in better calf performance.