Effects of evaporative cooling and dietary zinc source on heat shock responses and mammary gland development in lactating dairy cows during summer.

The objective of this study was to examine the effects of evaporative cooling and dietary supplemental Zn source on heat shock responses and mammary gland development of lactating dairy cows during summer. Seventy-two multiparous lactating Holstein cows were randomly assigned to 1 of 4 treatments in a 2 × 2 factorial arrangement. Cows were either cooled (CL) or not cooled (NC) and fed diets supplemented with 75 mg of Zn/kg of dry matter (DM) from Zn hydroxychloride (IOZ) or 35 mg of Zn/kg of DM from Zn hydroxychloride plus 40 mg of Zn/kg of DM from Zn-Met complex (ZMC). The 168-d trial included a 12-wk baseline phase when all cows were cooled and fed respective dietary treatments, and a subsequent 12-wk environmental challenge phase when NC cows were deprived of evaporative cooling. Plasma was collected from a subset of cows (n = 24) at 1, 3, 5, 12, 26, 41, 54, 68, 81 d of the environmental challenge to measure heat shock protein (HSP) 70 concentration. Mammary biopsies were collected from another subset of cows (n = 30) at enrollment (baseline samples) and at d 7 and 56 of the environmental challenge to analyze gene expression related to heat shock response, apoptosis and anti-oxidative enzymes, and to examine apoptosis and cell proliferation using immunohistochemistry. Supplemental Zn source did not affect milk yield but NC cows produced less milk than CL cows. Supplemental Zn source had no effect on mammary gene expression of HSP27, 70, and 90 or plasma concentrations of HSP70. The NC cows had greater mammary gene expression of HSP than CL cows. Circulating HSP70 of NC cows gradually increased and was higher at 81 d of environmental challenge compared with CL cows. Relative to IOZ, ZMC cows tended to have lower total mammary cell proliferation but greater mammary apoptosis. There was a tendency of greater TNFRSF1A mRNA expression for ZMC compared with IOZ cows, which may suggest upregulated extrinsic apoptosis. At d 7 of environmental challenge, NC cows had numerically higher mammary apoptosis than CL cows although not statistically significant. The NC cows tended to have greater mRNA expression of CAT and SOD3 regardless of time, and had greater mRNA expression of GPX1 at d 56 and FAS at d 7 of the environmental challenge than CL cows. Relative to CL cows, mammary cell proliferation rate was higher for NC cows at d 56 of the environmental challenge. In conclusion, dietary source of supplemental Zn has substantial effect on mammary cell turnover in lactating dairy cows, and prolonged exposure to heat stress increases mammary cell proliferation.

Effects of acute and chronic heat stress on feed sorting behaviour of lactating dairy cows.

Nutritional strategies to mitigate the negative effects of heat stress on animal welfare and productivity often involve changes in ration formulation. However, cattle commonly sort their ration in favour of certain components, and it is not clear how feed sorting responds to heat stress. This study investigated the association between heat stress and feed sorting behaviour. Lactating Holstein dairy cows (n = 32; parity = 2.8±1.2; mean±SD) were housed in a free stall barn and milked 3×/day. Cows were fed individually using the Calan Broadbent Feeding System and offered ad libitum access to a total mixed ration (containing on a dry matter basis: 3.3% ryegrass hay, 16.5% ryegrass baleage, 24.7% corn silage, 11.1% brewers grains, 19.7% ground corn, 19.8% concentrate and 4.9% protein/mineral supplement), provided 1×/day. Beginning at 186±60 days in milk, cows were exposed to either: heat stress conditions (HT; n = 15) (average temperature-humidity index: 77.6), or evaporative cooling (CL; n = 17), consisting of misters and fans over the freestall and feed bunks. Data were collected during a 4-day baseline period, and two 4-day experimental periods: starting at 10 days after implementing treatments (defined as acute heat stress for HT cows), and at 62 days after implementing treatments (defined as chronic heat stress for HT cows). Daily feed intake and physiological responses to heat stress (body temperature, respiration rate) were recorded. Samples of fresh and refused feed were collected daily from individual cows for particle size analysis. The particle size separator had three screens (19, 8 and 1.18 mm) and a bottom pan, resulting in 4 fractions (long, medium, short and fine particles). Feed sorting was calculated as the actual intake of each particle size fraction expressed as a percentage of the predicted intake of that fraction. During both heat stress periods, HT cows sorted for long particles more than CL cows (105.0% v. 100.6%; SE = 1.1). During acute heat stress, HT cows sorted to a greater extent than CL cows against medium and short particles, whereas sorting of these fractions did not differ during chronic heat stress. Body temperature and respiration rate were associated across treatments with the extent of sorting for long particles and against short particles during acute heat stress. These results suggest that feed sorting is particularly influenced during acute heat stress, and that sorting for longer particles may increase in heat stress.

Effects of heat stress and dietary zinc source on performance and mammary epithelial integrity of lactating dairy cows.

Dietary Zn and heat stress alter gut integrity in monogastric animals. However, effects of Zn on mammary epithelial integrity in heat-stressed lactating dairy cows have not been studied. Multiparous lactating Holstein cows (n = 72) were randomly assigned to 1 of 4 treatments with a 2 × 2 factorial arrangement to study the effects of environment and Zn source on performance and mammary epithelial integrity. Treatments included 2 environments [cooled (CL) or not cooled (NC)] and 2 Zn sources [75 mg/kg of supplemental Zn as Zn hydroxychloride (IOZ) or 35 mg/kg of Zn hydroxychloride + 40 mg/kg of Zn-Met complex (ZMC)]. The experiment was divided into baseline and environmental challenge phases of 84 d each. All cows were cooled during the baseline phase (temperature-humidity index = 72.5), whereas NC cows were not cooled during environmental challenge (temperature-humidity index = 77.7). Mammary biopsies were collected on d 7 and 56 relative to the onset of environmental challenge to analyze gene expression of claudin 1, 4, and 8, zonula occludens 1, 2, and 3, occludin, and E-cadherin and protein expression of occludin and E-cadherin. Deprivation of cooling increased respiration rate (64.8 vs. 73.9 breaths/min) and vaginal temperature (39.03 vs. 39.94°C) and decreased dry matter intake (26.7 vs. 21.6 kg/d). Energy-corrected milk yield decreased for NC cows relative to CL cows (24.5 vs. 34.1 kg/d). An interaction between environment and Zn source occurred for milk fat content as CL cows fed ZMC had lower milk fat percentage than other groups. Relative to CL cows, NC cows had lower concentrations of lactose (4.69 vs. 4.56%) and solids-not-fat (8.46 vs. 8.32%) but a higher concentration of milk urea nitrogen (9.07 vs. 11.02 mg/mL). Compared with IOZ, cows fed ZMC had lower plasma lactose concentration during baseline and tended to have lower plasma lactose concentration during environmental challenge. Plasma lactose concentration tended to increase at 3, 5, and 41 d after the onset of environmental challenge in NC cows relative to CL cows. Treatment had no effect on milk BSA concentration. Cows fed ZMC tended to have higher gene expression of E-cadherin relative to IOZ. Compared with CL, NC cows had increased gene expression of occludin and E-cadherin and tended to have increased claudin 1 and zonula occludens 1 and 2 gene expression in the mammary gland. Protein expression of occludin and E-cadherin was unchanged. In conclusion, removing active cooling impairs lactation performance and affects gene expression of proteins involved in the mammary epithelial barrier, and feeding a portion of dietary zinc as ZMC improves the integrity of the mammary epithelium.

Effects of feeding betaine-containing liquid supplement to transition dairy cows.

Betaine is a natural compound found in sugar beets that serves as a methyl donor and organic osmolyte when fed to animals. The objective was to evaluate the effect of feeding betaine-containing molasses on performance of transition dairy cows during late summer in 2 trials. In early September, cows were randomly assigned to betaine (BET) or control (CON) groups either shortly after dry off (trial 1; n = 10 per treatment) or 24 d before calving (trial 2; n = 8 per treatment) based on parity and previous mature equivalent milk yield. Cows were fed common diets supplemented either with a liquid supplement made of molasses from sugar cane and condensed beet solubles containing betaine [BET, 89.1 g/kg of dry matter (DM)] or a sugar cane molasses-based liquid supplement without betaine (CON) until 8 wk postpartum. The liquid supplements had similar nutrient contents and were fed at a rate of 1.1 and 1.4 kg DM/d for pre- and postpartum cows, respectively. Starting at their entry in the studies, cows were housed in the same freestall barn without a cooling system. After calving, all cows were housed in the same barn cooled by misters and fans and milked thrice daily. Intake was recorded daily and body weight and body condition score were assessed every 2 wk. Milk yield was recorded at each milking and composition was analyzed weekly. Blood samples were collected weekly from a subset of cows to assess concentrations of metabolites and AA. No treatment effects were apparent for DM intake and body weight in the prepartum and postpartum periods. For cows enrolled at dry off, BET supported higher milk yield (45.1 vs. 41.9 kg/d) and fat content (4.78 vs. 4.34%) and elevated plasma concentrations of nonesterified fatty acids and ß-hydroxybutyrate in early lactation compared to CON. However, no differences were observed for milk yield, most milk component contents and yields, and blood metabolites between treatments for cows enrolled during the close-up period. Compared to cows in the CON group, BET cows enrolled during the far-off period tended to have lower plasma concentrations of Met, Thr, and Trp during the pre- and postpartum periods. They also had lower plasma concentrations of Lys and Phe before calving but higher plasma Gly concentration after parturition. In conclusion, feeding a betaine-containing liquid supplement from far-off through early lactation improves lactation performance but increases adipose tissue mobilization and production of ketone bodies in early lactation.

In utero heat stress decreases calf survival and performance through the first lactation.

Calves born to cows exposed to heat stress during late gestation (i.e., the dry period) have lower birth weight and weaning weight and compromised passive immune transfer compared with those born to dams that are cooled. However, it is unknown if heat stress in utero has carryover effects after weaning. The objective was to evaluate the effect of heat stress (HT) or cooling (CL) in late gestation dairy cows on the survival, growth, fertility, and milk production in the first lactation of their calves. Data of animals obtained from previous experiments conducted during 5 consecutive summers in Florida were pooled and analyzed. Cows were dried off 46d before expected calving and randomly assigned to 1 of 2 treatments, HT or CL. Cooled cows were housed with sprinklers, fans, and shade, whereas only shade was provided to HT cows. Within 4h of birth, 3.8 L of colostrum was fed to calves from both groups of cows. All calves were managed in the same manner and weaned at 49d of age. Birth weight and survival of 146 calves (HT=74; CL=72) were analyzed. Additionally, body weight, growth rate, fertility, and milk production in the first lactation from 72 heifers (HT=34; CL=38) were analyzed. As expected, HT calves were lighter (means ± SEM; 39.1±0.7 vs. 44.8±0.7kg) at birth than CL calves. Cooled heifers were heavier up to 1yr of age, but had similar total weight gain (means ± SEM; 305.8±6.3 vs. 299.1±6.3kg, respectively) compared with HT heifers. No effect of treatment was observed on age at first insemination (AI) and age at first parturition. Compared with CL heifers, HT heifers had a greater number of services per pregnancy confirmed at d 30 after AI, but no treatment effect was observed on number of services per pregnancy confirmed at d 50 after AI. A greater percentage of CL heifers reached first lactation compared with HT heifers (85.4 vs. 65.9%). Moreover, HT heifers produced less milk up to 35wk of the first lactation compared with CL heifers (means ± SEM; 26.8±1.7 vs. 31.9±1.7kg/d), and no difference in body weight during lactation was observed (means ± SEM; HT: 568.4±14.3kg; CL: 566.5±14.3kg). These data suggest that heat stress during the last 6wk of gestation induces a phenotype that negatively affects survival and milk production up to and through the first lactation of offspring.

Short communication: Effect of maternal heat stress in late gestation on blood hormones and metabolites of newborn calves.

Maternal heat stress alters immune function of the offspring, as well as metabolism and future lactational performance, but its effect on the hormonal and metabolic responses of the neonate immediately after birth is still not clear. The objective of this study was to investigate the blood profiles of hormones and metabolites of calves born to cows that were cooled (CL) or heat-stressed (HS) during the dry period. Within 2 h after birth, but before colostrum feeding, blood samples were collected from calves [18 bulls (HS: n=10; CL: n=8) and 20 heifers (HS: n=10; CL: n=10)] born to CL or HS dry cows, and hematocrit and plasma concentrations of total protein, prolactin, insulin-like growth factor-I, insulin, glucose, nonesterified fatty acid, and ß-hydroxybutyrate were measured. Compared with CL, HS calves had lower hematocrit and tended to have lower plasma concentrations of insulin, prolactin, and insulin-like growth factor-I. However, maternal heat stress had no effect on plasma levels of total protein, glucose, fatty acid, and ß-hydroxybutyrate immediately after birth. These results suggest that maternal heat stress desensitizes a calf's stress response and alters the fetal development by reducing the secretion of insulin-like growth factor-I, prolactin, and insulin.

Effect of maternal heat stress during the dry period on growth and metabolism of calves.

Preliminary studies suggest that maternal heat stress (HS) during late gestation exerts carryover effects on a calf's insulin response after weaning, but a comprehensive evaluation of how maternal HS affects calf intake, growth, and metabolic response from birth to weaning is lacking. Our objective was to evaluate the effects of maternal HS during the dry period on dry matter intake, growth, and metabolism from birth to weaning. After birth, 20 heifers born to either HS (n=10) or cooled (CL, n=10) dry cows were immediately separated from their dams and fed 3.8 L of colostrum from a common pool within 4h of birth. All heifers were managed identically and weaned at 49 d of age (DOA). Calf starter intake was recorded daily, and body weight was assessed at birth and every 2 wk from birth to 56 DOA. Blood samples were collected twice a week until 56 DOA to assess hematocrit and concentrations of insulin and metabolites. To evaluate metabolic responses to maternal HS, a glucose tolerance test, insulin, and epinephrine challenge were performed on 3 consecutive days for all heifers at 8, 29, and 57 DOA. Maternal HS during the dry period did not affect heifer birth weight. Compared with HS, CL calves consumed more starter (0.53 vs. 0.34kg/d) from birth to 56 DOA and were heavier (71.7 vs. 61.4kg) at 56 DOA. Relative to HS calves, CL calves tended to have higher hematocrit (27.4 vs. 24.7%). No differences were found between treatments in plasma concentrations of insulin and glucose, but HS calves had higher nonesterified fatty acids and ß-hydroxybutyrate concentrations after 32 DOA. Compared with CL, HS calves had a faster glucose clearance after a glucose tolerance test and a slower insulin clearance after an insulin challenge. In conclusion, maternal HS during late gestation reduces calf starter intake and growth, alters blood metabolite profile, and increases noninsulin-dependent glucose uptake.

Effect of heat stress during late gestation on immune function and growth performance of calves: isolation of altered colostral and calf factors.

Calves born to cows exposed to heat stress during the dry period and fed their dams' colostrum have compromised passive and cell-mediated immunity compared with calves born to cows cooled during heat stress. However, it is unknown if this compromised immune response is caused by calf or colostrum intrinsic factors. Two studies were designed to elucidate the effects of colostrum from those innate to the calf. The objective of the first study was to evaluate the effect of maternal heat stress during the dry period on calf-specific factors related to immune response and growth performance. Cows were dried off 46 d before expected calving and randomly assigned to 1 of 2 treatments: heat stress (HT; n=18) or cooling (CL; n=18). Cows of the CL group were housed with sprinklers, fans and shade, whereas cows of HT group had only shade. After calving, the cows were milked and their colostrum was frozen for the subsequent study. Colostrum from cows exposed to a thermoneutral environment during the dry period was pooled and stored frozen (-20 °C). Within 4h of birth, 3.8L of the pooled colostrum from thermoneutral cows was fed to calves born to both HT and CL cows. Day of birth was considered study d 0. All calves were exposed to the same management and weaned at d 49. Blood samples were collected before colostrum feeding, 24h after birth and twice weekly up to d 28. Total serum IgG concentrations were determined. Body weight was recorded at birth and at d 15, 30, 45, and 60. Relative to CL calves, HT calves were lighter at birth (38.3 vs. 43.1 kg), but no difference in weight gain was observed at d 60. Additionally, HT calves had lower apparent efficiency of IgG absorption (26.0 vs. 30.2%), but no differences were observed for total IgG concentration. The objective of the second study was to evaluate the isolated effect of the colostrum from HT cows on calf immune response and growth performance. The experimental design was identical to the first study, but all calves were born to cows under thermoneutral conditions during the dry period. At birth, calves were blocked by sex and birth weight and then randomly assigned to 1 of 2 treatments, which meant they received pooled colostrum from HT cows or CL cows. No treatment effect was observed on passive immune transfer or on postnatal growth. Thus, heat stress during the last 6 wk of gestation negatively affects the ability of the calf to acquire passive immunity, regardless of colostrum source.

Short communication: Maternal heat stress during the dry period alters postnatal whole-body insulin response of calves.

Heat stress during the dry period not only negatively affects a cow's performance but also affects her offspring. Previous studies indicate that calves born to cows heat-stressed during late gestation have lower birth weight but similar overall weight gain during the prepubertal period compared with those cooled in utero. However, it is unclear if whole-body insulin response, and thus metabolism, of calves is altered in their postnatal life after in utero heat stress. The aim of the present study was to examine the effects of maternal heat stress during the dry period on whole-body insulin response of calves after weaning. Calves (10/treatment) were born to cows exposed to heat stress (HT) or cooling (CL) when dry. Calves were immediately separated from their dams and fed 3.8L of high-quality colostrum within 1h after birth and then 1.9L 12h later. All calves were fed 1.9 to 3.8L of pasteurized milk in the morning and afternoon from 2 to 42 d of age and then only in the morning until weaning at 49 d. Calf starter and water were offered ad libitum starting at 2 d of age. All calves were managed in the same manner throughout the study. All calves were subjected to a glucose tolerance test (GTT) and an insulin challenge (IC) at 55 d of age. Calves heat-stressed in utero were born lighter (40 ± 1.4 vs. 45 ± 1.4 kg) compared with CL calves. Both groups of calves had similar weaning weights (HT: 68 ± 3.2 kg; CL: 71 ± 3.3 kg) and body weight gain from birth to weaning (HT: 28 ± 2.2 kg; CL: 26 ± 2.3 kg). Compared with those cooled in utero, HT calves had a similar insulin response to GTT and insulin clearance during IC but faster glucose clearance during GTT and IC. In conclusion, in addition to impaired fetal growth, maternal heat stress during the dry period enhances the whole-body insulin response of calves after weaning, which suggests the possibility of accelerated lipogenesis and fat deposition in early life.

Prevalence of periparturient diseases and effects on fertility of seasonally calving grazing dairy cows supplemented with concentrates.

The objectives were to characterize the prevalence of periparturient diseases and their effects on reproductive performance of dairy cows in seasonal grazing farms. A total of 957 multiparous cows in 2 farms (555 in farm A and 402 in farm B) were evaluated and diseases characterized. At calving, dystocia, twin birth, stillbirth, and retained fetal membranes were recorded and grouped as calving problems. On d 7±3 and 14±3 postpartum, cows were evaluated for metritis and on d 28±3 for clinical endometritis based on scoring of the vaginal discharge. From parturition to 30 d after artificial insemination (AI), prevalence of mastitis, lameness, and digestive and respiratory problems were recorded. For subclinical diseases, diagnosis was based on blood samples collected from 771 cows and analyzed for concentrations of Ca, nonesterified fatty acids (NEFA), and ß-hydroxybutyrate. Cows were considered as having elevated NEFA concentration if the concentration was ≥0.70 mM, subclinical ketosis if the ß-hydroxybutyrate concentration was ≥0.96 mM, and subclinical hypocalcemia if the Ca concentration was ≤2.14 mM. Ovaries were scanned on d 35±3 and 49±3 postpartum for determination of estrous cyclicity. All cows were enrolled in a timed AI program and inseminated on the first day of the breeding season: on average, 86 d postpartum. Overall, 37.5% (359/957) of the cows presented at least 1 clinical disease and 59.0% (455/771) had at least 1 subclinical health problem. Prevalence of individual diseases was 8.5% for calving problems, 5.3% for metritis, 15.0% for clinical endometritis, 13.4% for subclinical endometritis, 15.3% for mastitis, 2.5% for respiratory problems, 4.0% for digestive problems, 3.2% for lameness, 20.0% for elevated NEFA concentration, 35.4% for subclinical ketosis, and 43.3% for subclinical hypocalcemia. Clinical and subclinical diseases had additive negative effects on reproduction, delaying resumption of estrous cyclicity and reducing pregnancy per AI (P/AI). Occurrence of multiple diseases further reduced reproductive efficiency compared with a single disease. Individually, subclinical hypocalcemia, elevated NEFA concentration, metritis, and respiratory and digestive problems reduced estrous cyclicity by d 49 postpartum. Elevated NEFA concentration, calving problem, metritis, clinical and subclinical endometritis, and digestive problems reduced P/AI on d 65 after AI. Moreover, calving problems and clinical endometritis increased the risk of pregnancy loss between gestation d 30 and 65. Serum concentrations of Ca and NEFA were negatively correlated, and both were associated with prevalence of uterine diseases. In conclusion, periparturient diseases were highly prevalent in seasonally calving grazing dairies and affected cows had delayed resumption of estrous cyclicity, reduced P/AI, and increased risk of pregnancy loss.