A.P.A. Monteiro

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 calfs 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 β-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.

Effects of late-gestation heat stress on immunity and performance of calves.

Lactating cows that experience heat stress will have reduced dry matter intake and milk yield and shift metabolism, which ultimately reduces the efficiency of milk production. Dry cows that are heat stressed similarly experience lower intake, reduced mammary growth, and compromised immune function that ultimately results in a poorer transition into lactation and lower milk yield in the next lactation. A recent focus in our laboratory is on the effects of late gestation, in utero heat stress on calf survival and performance. We have completed a series of studies to examine preweaning growth and health, and later reproductive and productive responses, in an attempt to quantify acute and persistent effects of in utero heat strain. Late gestation heat stress results in calves with lower body weight at birth, shorter stature at weaning, and failure to achieve the same weight or height at 12 mo of age observed in calves from dams that are cooled when dry. A portion of the reduced growth may result from the lower immune status observed in calves heat stressed in utero, which begins with poorer apparent efficiency of immunoglobulin absorption and extends to lower survival rates through puberty. Heat-stressed calves, however, have permanent shifts in metabolism that are consistent with greater peripheral accumulation of energy and less lean growth relative to those from cooled dams. Comparing reproductive performance in calves heat stressed versus those cooled in utero, we observe that the cooled heifers require fewer services to attain pregnancy and become pregnant at an earlier age. Tracking the milk production in calves that were heat stressed in utero versus those cooled in late gestation revealed a significant reduction of yield in the first lactation, approximately 5 kg/d through 35 wk of lactation, despite similar body weight and condition score at calving. These observations indicate that a relatively brief period of heat stress in late gestation dramatically alters the health, growth, and ultimate performance of dairy calves. Thus, it is critical to effectively manage heat stress of dry cows to avoid negative effects on the calf.

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 β-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 β-hydroxybutyrate immediately after birth. These results suggest that maternal heat stress desensitizes a calfs stress response and alters the fetal development by reducing the secretion of insulin-like growth factor-I, prolactin, and insulin.

Short communication: Effect of heat stress on markers of autophagy in the mammary gland during the dry period.

Heat stress (HT) during the dry period compromises mammary gland (MG) growth, thus negatively affecting subsequent milk yield. Cooling during the late dry period, when mammary tissue proliferates, is a common management practice. However, it neglects MG involution during the early dry period, a process that is accomplished by both apoptosis and autophagy. Our objective was to evaluate the effect of HT on MG autophagy during the early dry period. Holstein cows were dried off ~45d before expected calving and randomly assigned to 1 of 2 treatments: HT or cooling (CL). All cows were housed in the same free stall barn during the dry period, but only the stall area for CL cows was equipped with soakers and fans. Rectal temperature and respiration rate were measured daily during the dry period. Mammary gland biopsies were collected from each cow 3d before dry-off and on d 3, 7, 14, and 22±2 after dry-off. Autophagy in the MG was determined by measuring protein expression of 2 autophagic markers, autophagy-related protein 7 and microtubule-associated protein light chain 3 (LC3). The average temperature-humidity index during the dry period was 77.7, which indicated that HT and CL cows were exposed to significant heat stress. However, the cooling system effectively alleviated heat strain in CL cows by decreasing the rectal temperature (39.0 vs. 39.4°C) and respiration rate (47.3 vs. 71.2 breaths per minute) relative to HT cows. Protein expression of autophagy-related protein 7, a marker for early autophagosome formation, did not change within or between groups. In contrast, protein expression of LC3-II, a marker of autophagosomes, and its precursor LC3-I showed a dynamic expression pattern in MG from CL cows during the early dry period. Relative to HT cows, MG from CL cows displayed higher expression of LC3-I and LC3-II on d 7 and lower expression of LC3-II on d 14 and 22 after dry-off. Collectively, our data provide a possible mechanistic explanation for the impairment of MG capacity in HT dairy cows. Heat stress-related perturbations of autophagic activity may compromise the regenerative MG involution that is necessary for optimal cell proliferation.

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 β-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.

Short communication: Repeated mammary tissue collections during lactation do not alter subsequent milk yield or composition

Mammary biopsy collection (MB) is a valuable approach for studying mammary gland biology, but it is unclear if repeated MB impair the performance of lactating dairy cows. The objective of this trial was to examine the effect of repeated MB during lactation on udder health, dry matter intake (DMI), and lactation performance of lactating dairy cows. Sixty-four multiparous, mid-lactation Holstein cows were enrolled in a 29-wk trial, and 32 cows were randomly selected for repeated MB. The MB and non-MB (NMB) cows had similar parity (2.6 ± 0.9) and days in milk (96.5 ± 56.3 d) at enrollment. All animals were housed in the same barn and managed in the same manner. Cows were milked 3 times daily with milk yield recorded at each milking. Milk composition was measured weekly and DMI recorded daily. Three MB were performed per cow: 1 wk after enrollment and at 15 and 24 wk. The first and third MB were performed on the left rear quarter, whereas the second MB was on the right rear quarter. The MB were performed based on previously described procedures using a rotating stainless steel cannula with a retractable blade connected to a cordless drill, with appropriate sedation and antiseptic treatment after each MB. After MB, udder health, surgical wound healing, and presence of blood in milk were visually examined at each milking. Blood was cleared from milk 3.86 ± 2.0 d after MB. During the experiment, 4 rear quarters of MB cows and 5 rear quarters from NMB cows were diagnosed and treated for clinical mastitis. No differences were observed in DMI, milk yield, somatic cell score, or milk concentration and yields of fat, protein, lactose, and solids-not-fat between MB and NMB. In conclusion, lactating cows recover rapidly from MB, and repeated MB have no long-term effects on DMI, milk yield and composition, or udder health of lactating dairy cows.