Carryover effects of maternal late-gestation heat stress on granddaughters' growth and mammary gland development.

Maternal (F0) exposure to late-gestation heat stress reduces their daughter's (F1) mammary gland fat pad (FP) mass, parenchyma (PAR) mass, and epithelial cell proliferation when evaluated at birth and weaning, and the daughters go on to produce less milk in their first lactation. Herein, we investigated the effect of maternal late-gestation heat stress on whole-body growth and mammary development of their granddaughters (F2). Multiparous F0 cows had access to heat abatement (n = 41, shade, and active cooling via fans and water soakers) or not (n = 41, shade only) for the last 56 d of gestation during a subtropical summer. Consequently, the F1 daughters, born to F0 cows, were heat-stressed (HTF1, n = 36) or cooled (CLF1, n = 37) in utero during the last 2 mo of gestation. All F1 heifers were raised as an identically managed cohort until first calving. The F2 granddaughters, born to HTF1 (HTF2, n = 12) or CLF1 (CLF2, n = 17), were raised as an identically managed cohort until 70 d of age. Dry matter intake, BW, hip height, wither height, chest girth, head circumference, mammary gland teat length, and left-right and front-rear teat distances were measured. Average daily gain was calculated for the preweaning period (0-49 d). Mammary ultrasounds were performed on d 21, 49, and 70 (n = 9/group) on the rear left and right quarters to quantify PAR and FP areas. Mammary biopsies were collected for histological evaluation of epithelial structures (hematoxylin and eosin staining), and to quantify cells positive for estrogen receptor, α subunit (ERα), cell proliferation (Ki67), and apoptosis (terminal deoxynucleotidyl transferase dUTP nick-end labeling, TUNEL). Heifer growth from birth to d 49 was similar between CLF2 and HTF2 for all parameters evaluated. Distances between teats and teat length were not different between groups. On d 70, CLF2 heifers tended to have a greater average PAR (right and left quarters) relative to HTF2 heifers. Although the left FP was smaller in HTF2 heifers relative to CLF2 heifers, the average FP was not different. The lumenal and nonlumenal epithelial structures in the PAR of HTF2 heifers were significantly smaller than those of CLF2 heifers. In addition, HTF2 heifers had a reduced percentage of proliferating cells in the epithelial and stromal compartments and a greater percentage of apoptotic cells, particularly in the stroma. The percentage of ERα positive cells was significantly reduced in HTF2 heifers. In summary, although HTF2 heifers' DMI was similar and they grew at the same rate as CLF2 heifers throughout the preweaning phase, their mammary glands had smaller PAR areas with fewer epithelial structures characterized by reduced cell turnover and lower ERα expression. These early changes in the microstructure and cellular turnover of the mammary gland may partly explain the reduction in lactation performance relative to CLF2 counterparts at maturity.

Programming effects of intrauterine hyperthermia on adrenal gland development.

Maternal heat stress during late pregnancy can lead to intrauterine hyperthermia and affect fetal hypothalamic-pituitary-adrenal axis development and function. Herein, we investigated the effects of chronic environmental heat stress exposure of Holstein cows in the last 2 mo of gestation on their offspring's adrenal gland histomorphology and transcriptome. Cows in their last 54 ± 5 d of gestation were either heat stressed (housed under the shade of a freestall barn) or provided heat stress abatement via active cooling (via water soakers and fans) during a subtropical summer (temperature-humidity index >68). Respiration rate (RR) and skin temperature (ST) were elevated in heat-stressed dams relative to the cows with access to heat abatement (23 breaths/min and 2°C higher for RR and ST, respectively). Heifers born to heat-stressed cows experienced heat stress in utero (HS), whereas heifers born to actively cooled cows did not (CL). The adrenal gland was harvested from 6 heifers per group that were euthanized at birth (d 0; n = 12) or 1 wk after weaning (d 63; n = 12). Circulating cortisol was measured from blood samples collected weekly throughout the preweaning period. At d 63, heifers that experienced HS while developing in utero had heavier adrenal glands, with a greater total tissue surface area and thickness of the zona glomerulosa (ZG), fasciculata (ZF), and reticularis (ZR), compared with CL heifers. In addition, the adrenal gland of HS heifers had fewer cells in the ZG, more and larger cells in the ZF, and larger cells in the ZR, relative to CL heifers. Although no changes in circulating cortisol were observed through the preweaning period, the transcriptomic profile of the adrenal tissue was altered by fetal exposure to hyperthermia. Both at birth and on d 63, approximately 30 pathways were differentially expressed in the adrenal glands of HS heifers relative to CL. These pathways were associated with immune function, inflammation, prolactin signaling, cell function, and calcium transport. Upstream regulators significantly activated or inhibited in the adrenal glands of heifers exposed to intrauterine hyperthermia were identified. Maternal exposure to heat stress during late gestation caused an enlargement of their offspring's adrenal glands by inducing ZG and ZF cell hypertrophy, and caused gene expression changes. These phenotypic, histological, and molecular changes in the adrenal gland might lead to alterations in stress, immune, and metabolic responses later in life.

Long-term growth, feed efficiency, enteric methane emission, and blood metabolite responses to in utero hyperthermia in Holstein heifers.

Dairy producers are experiencing production and animal welfare pressures from the increasing frequency and severity of heat stress events due to global climate change. Offspring performance during the preweaning and lactating periods is compromised when exposed to heat stress during late gestation (in utero). However, knowledge of the lingering effects of in utero heat stress on yearling dairy heifers is limited. Herein, we investigated the long-term effects of in utero heat stress on heifer growth, feed efficiency, and enteric methane emissions in postpubertal heifers. During the last 56 d of gestation, 38 pregnant cows carrying heifer calves were exposed to either heat stress (IUHT; n = 17) or artificial cooling (IUCL; n = 21). At 18 ± 1 mo of age, the resulting IUCL and IUHT heifers were enrolled in the present 63-d study. Heifers were blocked by weight and randomly assigned to 3 pens with Calan gates. Body weights were recorded on 3 consecutive days at the start and end of the trial and used to calculate ADG. Body condition score, hip width, body length, and chest girth were measured at the start and end of the study. All heifers were fed a TMR comprised of 46.6% oatlage, 44.6% grass/alfalfa haylage, 7.7% male-sterile corn silage, 0.3% urea, and 0.8% mineral/vitamin supplement (on a DM basis). The TMR and refusal samples were obtained daily, composited weekly, and dried to calculate DMI. During the study, each pen had access to a GreenFeed unit for 8 ± 1d to measure CH4 and CO2 gas fluxes. During the last 3 d of measuring CH4 and CO2 fluxes, fecal samples were collected, composited by animal, dried, and analyzed to calculate NDF, OM, and DM digestibility. On the last day of fecal sampling, blood samples were also collected via coccygeal venipuncture, and GC time-of-flight MS analysis was performed. Residual feed intake (RFI; predicted DMI - observed DMI), and feed conversion efficiency (FCE; DMI/ADG) were calculated to estimate feed efficiency. No differences were found in initial or final BW, hip width, chest girth, or BCS; however, IUCL heifers were longer in body length compared with IUHT heifers. Dry matter intake, ADG, RFI, and FCE were similar between IUHT and IUCL heifers. In utero heat-stressed and IUCL heifers produced similar amounts of CH4 and CO2, and no differences were found in the number of GreenFeed visits or latency to approach the GreenFeed. The concentrations of 6 blood metabolites involved in lipogenic pathways were different between in utero treatments. In conclusion, in utero heat stress does not seem to have long-term effects on feed efficiency or methane emissions during the postpubertal growing phase; however, IUCL heifers maintained a body-length advantage over their IUHT counterparts and differed in concentrations of several candidate metabolites that encourage further exploration of their potential function in key organs, such as the liver and mammary gland.

Intravenous infusion of 5-hydroxytryptophan to mid-lactation Holstein cows transiently affects milk production and circulating amino acid concentrations.

In dairy cows, the lactating mammary glands synthesize serotonin, which acts in an autocrine-paracrine manner in the glands and is secreted into the periphery. Serotonin signaling during lactation modulates nutrient metabolism in peripheral tissues such as adipose and liver. We hypothesized that the elevation of circulating serotonin during lactation would increase nutrient partitioning to the mammary glands, thereby promoting milk production. Our objective was to elevate circulating serotonin via intravenous infusion of the serotonin precursor 5-hydroxytryptophan (5-HTP) to determine its effects on mammary supply and extraction efficiency of AA, and milk components production. Twenty-two multiparous mid-lactation Holstein cows were intravenously infused with 5-HTP (1 mg/kg body weight) or saline, in a crossover design with two 21-d periods. Treatments were infused via jugular catheters for 1 h/d, on d 1 to 3, 8 to 10, and 15 to 17 of each period, to maintain consistent elevation of peripheral serotonin throughout the period. Milk and blood samples were collected in the last 96 h of each period. Whole-blood serotonin concentration was elevated above saline control for 96 h after the last 5-HTP infusion. Dry matter intake was decreased for cows receiving 5-HTP, and on average they lost body weight over the 21-d period, in contrast to saline cows who gained body weight. Milk production and milk protein yield were lower in cows receiving 5-HTP during the 3 infusion days, but both recovered to saline cow yields in the days after. Although milk fat yield exhibited a day-by-treatment interaction, no significant difference occurred on any given day. Milk urea nitrogen concentration was lower in 5-HTP cows on the days following the end of infusions, but not different from saline cows on infusion days. Meanwhile, plasma urea nitrogen was not affected by 5-HTP infusion. Circulating concentrations of AA were overall transiently decreased by 5-HTP, with concentrations mostly returning to baseline within 7 h after the end of 5-HTP infusion. Mammary extraction efficiency of AA was unaffected by 5-HTP infusion. Overall, both lactation performance and circulating AA were transiently reduced in cows infused with 5-HTP, despite sustained elevation of circulating serotonin concentration.

Effects of thermal stress on calf welfare.

Cold and heat stress present welfare challenges for dairy calves. The consequences of thermal stress on biological functioning have been well documented, and many housing and management strategies have been evaluated to mitigate those detrimental impacts. In cold weather, mitigation strategies have largely focused on nutritional interventions or limiting heat loss with resources such as bedding or jackets. In hot weather, heat abatement strategies such as supplemental shade, increased environmental air exchange through passive ventilation, and forced air movement through mechanical ventilation have been evaluated. Recently in Wisconsin's continental climate, our group evaluated how 2 aspects of calf welfare-the needs for thermal comfort and social contact (i.e., pair or group housing vs. individual housing)-may align or conflict in winter and summer, respectively. In both seasons, calves pair-housed in outdoor hutches preferred social proximity. When 2 calves shared a hutch, the heat load was greater than for a single calf, which may be beneficial for thermal comfort in winter. In summer, the potential detriments from the additional heat load of 2 calves was mitigated with passive hutch ventilation, which calves preferred. Nonetheless, knowledge gaps remain regarding the impacts of thermal stress on calves' affective states, and much remains unknown about their preferences and motivations for specific thermal stress mitigation resources. Future research to address these gaps could improve our understanding of calf welfare and inform best practices for calf management.

Exploring the contribution of mammary-derived serotonin on liver and pancreas metabolism during lactation.

During lactation, the murine mammary gland is responsible for a significant increase in circulating serotonin. However, the role of mammary-derived serotonin in energy homeostasis during lactation is unclear. To investigate this, we utilized C57/BL6J mice with a lactation and mammary-specific deletion of the gene coding for the rate-limiting enzyme in serotonin synthesis (TPH1, Wap-Cre x TPH1FL/FL) to understand the metabolic contributions of mammary-derived serotonin during lactation. Circulating serotonin was reduced by approximately 50% throughout lactation in Wap-Cre x TPH1FL/FL mice compared to wild-type mice (TPH1FL/FL), with mammary gland and liver serotonin content reduced on L21. The Wap-Cre x TPH1FL/FL mice had less serotonin and insulin immunostaining in the pancreatic islets on L21, resulting in reduced circulating insulin but no changes in glucose. The mammary glands of Wap-Cre x TPH1FL/FL mice had larger mammary alveolar areas, with fewer and smaller intra-lobular adipocytes, and increased expression of milk protein genes (e.g., WAP, CSN2, LALBA) compared to TPH1FL/FL mice. No changes in feed intake, body composition, or estimated milk yield were observed between groups. Taken together, mammary-derived serotonin appears to contribute to the pancreas-mammary cross-talk during lactation with potential implications in the regulation of insulin homeostasis.

Actively ventilating calf hutches using solar-powered fans: Effects on hutch microclimate and calf thermoregulation.

Although active ventilation via fans is an effective and widely adopted heat abatement method for use with adult dairy cattle, it has yet to be investigated in outdoor hutch-housed dairy calves despite most US calves being raised in such systems. We investigated a solar-powered fan system for outdoor calf hutches and its effect on hutch microclimate and calf thermoregulation. During summer, a 3 × 3 Latin square was replicated 4 times (n = 12 preweaning heifers) with 4-d exposure periods to minimally (CON; rear windows closed), passively (PASS; rear windows opened), or actively (ACT; solar-powered fan, activated at dry bulb temperature [Tdb] > 21°C) ventilated hutch systems. Hutch microclimate and calf thermoregulation were evaluated either continuously (Tdb, humidity, rectum surface temperature, and behavior) or after a daily 30-min inside restriction (air speed, air particle number, noise level, respiration, and sweating rate, and skin and rectal temperature). Active ventilation substantially increased hutch air speed relative to PASS and CON (1.76 vs. 0.19 vs. 0.05 m/s). However, PASS hutches had the lowest INT Tdb (27.2 vs. 26.4 vs. 27.8°C), whereas ACT INT Tdb was reduced at 0900 and 1000 h relative to CON but not PASS. Similarly, ACT reduced calf respiration rates and lowered rectum surface temperature at 0800 and 0900 h when compared with CON but not PASS. The lack of strong ACT influence on calf outcomes over PASS could partially be explained by the decreased proportion of time ACT calves spent inside their hutch (48.7 vs. 67.3 vs. 64.1% of each hour). Overall, ACT improved hutch microclimate and calf responses relative to CON but not PASS. Either ACT or PASS ventilation may be sufficient to provide heat abatement to continental hutch-housed calves.