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 (i.e., housed under the shade of a free stall barn) or provided heat-stress abatement via active cooling (i.e., 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 bpm and 2 ◦C higher for RR and ST, respectively). Heifers born to heat-stressed cows experienced heat stress in utero (HS), while 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 one week after weaning (d 63; n = 12). Circulating cortisol was measured from blood samples collected weekly throughout the pre-weaning 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 in utero HS heifers had less 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 pre-weaning 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 in utero 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.

Prepartum heat stress in dairy cows increases postpartum inflammatory responses in blood of lactating dairy cows.

Uterine diseases and heat stress (HS) are major challenges for the dairy cow. Heat stress alters host immune resilience, making cows more susceptible to the development of uterine disease. Although HS increases the incidence of uterine disease, the mechanisms by which this occurs are unclear. We hypothesize that evaporative cooling (CL) to alleviate HS in prepartum cows has carry-over effects on postpartum innate immunity. Nulliparous pregnant Holstein heifers were assigned to receive either forced CL that resulted in cool conditions (shade with water soakers and fans; n = 14) or to remain under HS conditions (barn shade only; n = 16) for 60 d prepartum. Postpartum, all cows were housed in a freestall barn equipped with shade, water soakers, and fans. Respiratory rate and rectal temperature during the prepartum period were greater in HS heifers compared with CL heifers, indicative of HS. Although milk production was decreased in HS cows compared with CL cows, the incidence of uterine disease and content of total or pathogenic bacteria in vaginal mucus on d 7 or d 21 postpartum was not affected by treatment. Whole blood was collected on d 21 and subjected to in vitro stimulation with lipopolysaccharide. Lipopolysaccharide-induced accumulation of IL-1ß, IL-10, and MIP-1α was greater in blood collected from HS cows compared with CL cows. Our results imply that prepartum HS during late pregnancy has carry-over effects on postpartum innate immunity, which may contribute to the increased incidence of uterine disease observed in cows exposed to prepartum HS.

Use of a commercial feed supplement based on yeast products and microalgae with or without nucleotide addition in calves.

The use of feed additives with antioxidant and immune response modulatory activity could be a useful strategy in suckling calves to reduce morbidity and mortality. This strategy is based on several feed additives tested for these purposes. The aim of the paper is the examination of a commercial feed additive for adult cows for use in calves, with and without nucleotide supplementation. Seventy-five Holstein Friesian male calves were divided in 3 groups, with each calf randomly assigned to a group according to birth order. All calves received 2 L of pooled colostrum within 2 h of birth. The commercial feed supplement group was orally administered with 5 g/head of Decosel (dried brewer's yeast lysate (Saccharomyces cerevisiae), brewer's yeast walls (Saccharomyces cerevisiae), diatoms, spirulina, barley flour, calcium carbonate; Agroteam srl, Torrimpietra, Italy) and the nucleotides + commercial feed supplement group was orally administered with 5 g/head of an additive containing 2.5 g of Decosel and 2.5 g of nucleotides once daily from birth to 25 d. The control group was orally administered 20 mL of fresh water/head once daily. Calves that received the supplement and the nucleotides showed lower rates of protein and metabolizable energy conversion, with longer villi and greater crypt depth in duodenum. Moreover, the commercial feed supplement alone increased antioxidant capacity [2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and ferric-reducing antioxidant power] in plasma some activity of antioxidant liver enzymes, and peripheral blood mononuclear cell viability after in vitro concanavalin A and H2O2 stimuli. Dietary supplementation with a commercial feed supplement containing yeast products (yeast cell walls and hydrolyzed yeast) and microalgae enhanced the redox balance and gut morphology in calves, allowing calves to improve their immune response, increasing resistance to stress. Moreover, these beneficial effects were strongly potentiated when dietary nucleotides were added to the supplement.

MILK Symposium Introduction: Dairy production in developing countries.

In low- and middle-income countries (LMICs), dairy production is highly valued, and demand for milk is projected to continue to increase markedly over the next few decades. This presents a tremendous opportunity to improve the nutrition, health, incomes, and livelihoods of millions of people with the high-quality protein and bioavailable micronutrients in dairy products. However, low dairy consumption levels, due to low affordability, accessibility, and availability, still typify several LMICs. This is caused by inadequate feeding, management, and genetics; poor transport, cooling, and processing infrastructure; unconducive policy environments; and sociocultural and demographic factors. Strategies to address some of these factors were presented at the MILK Symposium hosted by the Feed the Future Innovation Lab for Livestock Systems during the 2019 American Dairy Science Association Annual Meeting. The papers presented are full manuscripts in this Special Issue of the Journal of Dairy Science. They address the importance of dairy products for human health, strategies to address feed, management, health, and food safety challenges in dairy production systems, and sustainability of dairy production in LMICs. They collectively show how strategic interventions can lead to marked improvements in dairy production in developing countries. These will ultimately contribute to meeting the growing global demand for milk and to achievement of the United Nations Sustainable Development Goals.

MILK Symposium review: Improving control of mastitis in dairy animals in Nepal.

Dairy animals are an important source of income, food, and nutritional security, and improvements in the productivity of dairy animals substantially improve the wellbeing of smallholder dairy farmers. As in other developing countries, dairy animals are key for rural livelihoods in Nepal but often suffer from mastitis-a production disease causing economic losses to farmers, challenges to the dairy processing industry, and possible health hazards to consumers. Studies show that the prevalence of subclinical mastitis in Africa and Asia typically exceeds 50%, threatening animal wellbeing, farmers, dairy processors, and consumers. We conducted a study in Nepal to develop a technology training package to control mastitis in dairy animals. Following identification of knowledge gaps, a technology package consisting of (1) developing good husbandry practices, implementing mastitis detection and control technologies; and (2) training technicians and farmers was implemented. A strategy was subsequently established to provide feedback to farmers in dairy cooperatives on the subclinical mastitis status of their cows. The package was applied in the mid-western region of Nepal. Six months after implementation, we observed a reduction in subclinical mastitis prevalence: from 55% (baseline) to 28% (endline; n = 432) in dairy cows and from 78% to 18% (n = 216) in buffalo. These positive study outcomes strongly suggest that the mastitis technology training package should be scaled across smallholder farmers within and beyond Nepal to control mastitis in dairy animals.

MILK Symposium review: Identifying constraints, opportunities, and best practices for improving milk production in market-oriented dairy farms in Sri Lanka.

Dairy is the most important subsector in the Sri Lankan livestock industry, due to the need to address the growing demand for fresh milk and milk products, and because of its potential influence on the rural economy. The USDA Food for Progress program awarded a 4.5-year Market-Oriented Dairy project to International Executive Service Corps, a not-for-profit organization based in Washington, DC. The objective of the Market-Oriented Dairy project is to support Sri Lanka's dairy sector and catalyze sustainable growth by strengthening the dairy sector through better technological, financial, and management practices benefiting all stakeholders and consumers along the dairy value chain. The University of Florida is working with International Executive Service Corps as technical experts in conducting dairy value chain assessments, identifying gaps and challenges in dairy management practices, extension services, milk quality management standards, and artificial insemination services. Assessment of the dairy value chain in 2018 identified a lack of good quality and quantity of feed, along with poor dairy management practices and ineffective extension services as major constraints to improving dairy productivity in Sri Lanka. In addition, lack of national milk quality standards that are consistent with international benchmarks and inadequate cooling facilities are significant challenges to improving milk quality. The nutritional status of cows is not suitable for optimal reproductive performance, compromising the success of artificial insemination in Sri Lanka. Based on these findings, we developed a dairy assessment tool and provided comprehensive training sessions targeting extension agents, veterinarians, and farmers to promote best practices in dairy management. Beyond training, however, industry support for standardization and monitoring of milk and feed quality are needed, providing opportunities for private investment to support the dairy industry. Similar opportunities are available for forage production and delivery to producers. The broader aim of the Market-Oriented Dairy project intervention is to reduce Sri Lanka's dependency on imported milk and contribute toward the goal of a safe, self-sufficient fresh milk supply.

Effect of heat stress during the early and late dry period on mammary gland development of Holstein dairy cattle.

Dry period heat stress impairs subsequent milk yield. Our objective was to evaluate the effect of heat stress or cooling during the early and late dry period on mammary gland gene expression and microstructure. Cows were dried off ∼45 d before expected parturition and randomly assigned to 1 of 2 treatments: heat stress (HT, n = 39) or cooling (CL, n = 39) during the first 21 d of the dry period. On d 22, cows were switched or remained on HT and CL and this yielded 4 treatments: heat stress during the entire dry period (HTHT, n = 18); cooling during the entire dry period (CLCL, n = 20); HT for the first 21 d dry, then CL until calving (HTCL, n = 21); or CL for the first 21 d dry, then HT until calving (CLHT, n = 19). Data were analyzed in 2 periods: first 21 d dry (early dry period) and from 22 d until calving (late dry period) and analyzed using PROC MIXED or GLM in SAS (SAS Institute Inc., Cary, NC). Mammary biopsies (5-8 cows/treatment) were collected at -3, 3, 7, 14, and 25 d relative to dry-off to evaluate mammary gland gene expression and histology [i.e., cellular apoptosis (terminal deoxynucleotidyl transferase dUTP nick end labeling) and proliferation (Ki67)]. Mammary alveoli number and connective tissue were visualized by hematoxylin and eosin and Mason's trichrome staining, respectively. During the early dry period, CL upregulated expression of CASP3, IGF1R, HSP90, HSF1, BECN1, ATG3, ATG5, and PRLR-LF relative to HT. However, in the late dry period, CLHT treatment upregulated expression of CASP3, CASP8, HSP70, HSP90, PRLR-LF, STAT5, CSN2, and ATG3 relative to CLCL. During the early dry period, cows exposed to HT had reduced mammary and stroma cell apoptosis and proliferation relative to CL. In addition to these findings, cows exposed to HT had lower connective tissue 3 d after dry-off relative to CL. However, in the late dry period, HTHT cows had higher connective tissue relative to CLCL. Also, in the early dry period, cows exposed to HT had greater alveoli number relative to CL, and HT decreased expression of genes related to autophagy and apoptosis in the early dry period, consistent with a delay in involution with HT. Thus, cows exposed to HT have extended involution with delayed apoptosis and autophagy signaling. Also, HT compromises mammary gland cell proliferation and leads to higher connective tissue later in the dry period. These results provide evidence that heat stress impairs overall mammary gland turnover during the dry period, which then affects secretory activity and productivity in the next lactation.

Effect of heat stress during early, late, and entire dry period on dairy cattle.

Cooling during the entire dry period abates the negative effects of heat stress postpartum, yet the temporal relationship of cooling (i.e., early or late dry period) to performance is unknown. We evaluated the effect of heat stress early, late, and for the entire dry period on subsequent performance. Cows were selected based on mature-equivalent milk yield and dried off 45 d before expected calving. Cows were blocked by parity, previous 305-d mature equivalent milk yield, and body weight (BW) and randomly assigned to cooling (shade, fans, and soakers; CL) or heat stress (shade; HT). Treatments included CL (n = 20) or HT (n = 18) during the entire dry period, HT during the first 3 wk dry and then CL until calving (HTCL, n = 21), or CL during the first 3 wk dry period and then HT until calving (CLHT, n = 19). Heat stress increased rectal temperature (RT; CL, 38.8; HT, 39.1 ± 0.04°C) and respiration rate (RR; CL, 52.9; HT, 70.5 ± 1.9 breaths/min) during the early dry period. In the late dry period, HT increased RT and RR relative to CL cows (RT = CL, 38.7; HT, 39.1; CLHT, 39.1; HTCL, 38.9 ± 0.05°C; RR = CL, 47; HT, 64; CLHT, 66; HTCL, 53 ± 2.1 breaths/min). During the early dry period, HT decreased dry matter intake (CL, 11.8; HT, 10.5 ± 0.35 kg/d) but dry matter intake did not differ among treatments during late dry period (HT, 10.7; HTCL, 11.1; CL, 11.2; CLHT, 10.1 ± 0.55 kg/d). Cows exposed to prepartum cooling during the entire dry period had increased dry matter intake compared with cows exposed to heat stress during the late dry period (CL vs. CLHT, 11.2 ± 0.55 and 10.1 ± 0.55 kg/d, respectively). Heat stress at any time reduced gestation length compared with cows under prepartum cooling during the entire dry period (CL, 277 vs. HT, 274; CLHT, 273; and HTCL, 274 ± 1.17 d). Dry period length decreased by approximately 4 d if cows were exposed to HT at any time. During the early dry period, HT decreased BW, whereas CL increased BW relative to that at dry-off (CL, 6.9; HT, -9.4 ± 3.7 kg). In the late dry period, we detected no differences in BW gain among treatments, but cows exposed to prepartum cooling for the entire dry period tended to have increased BW gain compared with HT and HTCL. Prepartum cooling during the early or late dry period alone partially rescued milk yield only in the first 3 wk of lactation (CL, 32.9; HT, 26.6; CLHT, 29.7; HTCL, 30.7 ± 1.37 kg/d). Cooling for the entire dry period increased milk yield up to 30 wk into lactation compared with all other treatments. Thus, HT at any time during the dry period compromises performance of cows after calving.

Liver proteomic analysis of postpartum Holstein cows exposed to heat stress or cooling conditions during the dry period.

Heat stress negatively affects cow performance, compromises immune function, and increases susceptibility to metabolic disorders, particularly during the dry period and as cows transition from gestation to lactation. Metabolic adaptations of the liver are critical for successful transition, yet it is unclear how heat stress affects metabolic pathways within the liver at the proteomic level. The objective of this study was to investigate the liver proteome of postpartum cows that were cooled or heat stressed during the dry period to gain insight into how protein expression is altered by prior heat stress and may contribute to performance and disease outcomes. During the dry period, cows were either housed in shaded barns with fans and water soakers [cooled group (CL); n = 5] or in shaded barns lacking these cooling devices [heat-stressed group (HT); n = 5]. Liver biopsies were collected at 2 d postpartum, and protein content was analyzed by label-free quantitative shotgun proteomics (nanoscale liquid chromatography coupled to tandem mass spectrometry). In the most comprehensive bovine liver proteomics analysis completed to date, we identified 3,270 proteins, 75 of which were differentially expressed between HT and CL cows (fold change ±1.2). The top pathways differing between HT and CL cows were oxidative phosphorylation, mitochondrial dysfunction, farnesoid X receptor/retinoid X receptor (FXR/RXR) activation, and the methylmalonyl pathway. Cooling cows during the dry period likely improves ATP production, reduces oxidative stress, and prevents excessive accumulation of hepatic triglycerides and cholesterol, which may contribute to greater milk yield and lower susceptibility to transition-related diseases.

Effects of feeding an immunomodulatory supplement to heat-stressed or actively cooled cows during late gestation on postnatal immunity, health, and growth of calves.

Heat stress during late gestation negatively affects the physiology, health, and productivity of dairy cows as well as the calves developing in utero. Providing cows with active cooling devices, such as fans and soakers, and supplementing cows with an immunomodulating feed additive, OmniGen-AF (OG; Phibro Animal Health Corporation), improves immune function and milk yield of cows. It is unknown if maternal supplementation of OG combined with active cooling during late gestation might benefit the developing calf as well. Herein we evaluated markers of innate immune function, including immune cell counts, acute phase proteins, and neutrophil function, of calves born to multiparous dams in a 2 × 2 factorial design. Dams were supplemented with OG or a bentonite control (NO) beginning at 60 d before dry off and exposed to heat stress with cooling (CL) or without active cooling (HT) during the dry period (∼46 d). At birth, calves were separated from their dams and fed 6.6 L of their dams' colostrum in 2 meals. Calf body weight and rectal temperature were recorded, and blood samples were collected at birth (before colostrum feeding) and at 10, 28, and 49 d of age. Calves born to either CL dams or OG dams were heavier at birth than calves born to HT or NO dams, respectively. Concentrations of serum amyloid A were higher in the blood of calves born to OG dams relative to NO and for HT calves relative to CL calves. In addition, calves born to cooled OG dams had greater concentrations of plasma haptoglobin than calves born to cooled control dams. Neutrophil function at 10 d of age was enhanced in calves born to cooled OG dams and lymphocyte counts were higher in calves born to OG dams. Together these results suggest that adding OG to maternal feed in combination with active cooling of cows during late gestation is effective in mitigating the negative effects of in utero heat stress on postnatal calf growth and immune competence.

Effect of nutritional immunomodulation and heat stress during the dry period on subsequent performance of cows.

Heat stress in dairy cows during the dry period impairs milk yield in the next lactation. Feeding OmniGen-AF (OG; Phibro Animal Health Corp., Teaneck, NJ) to lactating cows during heat stress may increase dry matter intake (DMI) and lowers respiration rate (RR) and rectal temperature (RT), but the effects in dry cows are not known. We hypothesized that OG supplementation before, during, and after the dry period (approximately 160 d total) would overcome the effects of heat stress and improve cow performance in the next lactation. Cows were randomly assigned to OG or control (placebo) treatments for the last 60 d in milk (DIM), based on mature-equivalent milk yield in the previous lactation. Cows were dried off 45 d before expected calving and randomly assigned to heat stress (HT) or cooling (CL) treatments. Thus, cows received dietary supplementation during late lactation before they were exposed to either CL or HT. After dry-off, treatment groups included heat stress with placebo (HT, only shade, 56 g/d of placebo, n = 17), HT with OG supplementation (HTOG, 56 g/d of OG, n = 19), cooling with placebo (CL, shade, fans, and soakers, 56 g/d of placebo, n = 16), and CL with OG supplementation (CLOG, 56 g/d of OG, n = 11). After parturition, all cows were kept under the same CL system and management, and all cows continued to receive OG or control treatment until 60 DIM. Cooling cows during the dry period reduced afternoon RT (CL vs. HT; 38.9 ± 0.05 vs. 39.3 ± 0.05°C) and RR (CL vs. HT; 45 ± 1.6 vs. 77 ± 1.6 breaths/min). Respiration rate was also decreased by OG supplementation under HT conditions (HTOG vs. HT; 69.7 ± 1.6 vs. 77.2 ± 1.6 breaths/min). An interaction was observed between OG supplementation and HT; HTOG cows tended to have lower morning RT compared with HT cows. During the dry period, OG reduced DMI relative to control cows. Birth weight was greater in calves from CL cows (CL vs. HT; 40.6 ± 1.09 vs. 38.7 ± 1.09 kg). No differences were detected among treatments in hematocrit, total protein, and body condition score. Cows offered CLOG, CL, and HTOG treatments had greater body weight during the dry period (794.9 ± 17.9, 746.8 ± 16.7, and 762.9 ± 14.9 kg, respectively) than HT cows (720 ± 16.2 kg). Gestation length was approximately 4 d longer for CL cows compared with HT cows. Cows offered CLOG, CL, and HTOG treatments produced more milk (41.3 ± 1.6, 40.7 ± 1.6, and 40.5 ± 1.6 kg/d, respectively) than HT treatment (35.9 ± 1.6 kg/d). Body weight after parturition and DMI were evaluated up to 60 DIM and averaged 661.5 ± 15.8 and 19.4 ± 0.7 kg/d, respectively, with no differences observed among treatments. These results confirm that exposure of dry cows to heat stress negatively affects milk yield in the subsequent lactation. Active cooling of dry cows and OG supplementation can reduce the negative effects of heat stress in the dry period on subsequent performance.

Elevated maternal cortisol leads to relative maternal hyperglycemia and increased stillbirth in ovine pregnancy.

In normal pregnancy, cortisol increases; however, further pathological increases in cortisol are associated with maternal and fetal morbidities. These experiments were designed to test the hypothesis that increased maternal cortisol would increase maternal glucose concentrations, suppress fetal growth, and impair neonatal glucose homeostasis. Ewes were infused with cortisol (1 mg·kg(-1)·day(-1)) from day 115 of gestation to term; maternal glucose, insulin, ovine placental lactogen, estrone, progesterone, nonesterified free fatty acids (NEFA), ß-hydroxybutyrate (BHB), and electrolytes were measured. Infusion of cortisol increased maternal glucose concentration and slowed the glucose disappearance after injection of glucose; maternal infusion of cortisol also increased the incidence of fetal death at or near parturition. The design of the study was altered to terminate the study prior to delivery, and post hoc analysis of the data was performed to test the hypothesis that maternal metabolic factors predict the fetal outcome. In cortisol-infused ewes that had stillborn lambs, plasma insulin was increased relative to control ewes or cortisol-infused ewes with live lambs. Maternal cortisol infusion did not alter maternal food intake or plasma NEFA, BHB, estrone, progesterone or placental lactogen concentrations, and it did not alter fetal body weight, ponderal index, or fetal organ weights. Our study suggests that the adverse effect of elevated maternal cortisol on pregnancy outcome may be related to the effects of cortisol on maternal glucose homeostasis, and that chronic maternal stress or adrenal hypersecretion of cortisol may create fetal pathophysiology paralleling some aspects of maternal gestational diabetes.

In utero hyperthermia in late gestation derails dairy calf early-life mammary development.

Prenatal hyperthermia has immediate and long-term consequences on dairy cattle growth, immunity, and productivity. While changes in the molecular architecture are reported in the mature mammary gland (MG), any influence on early-life mammary development is unknown. Herein, we characterize the impact of late-gestation in utero heat stress on heifer mammary gross and cellular morphology at early-life developmental stages (i.e., birth and weaning). During summer, pregnant dams were exposed to environmental heat stress (shade of a free-stall barn) or offered active cooling (shade, fans, and water soakers) for 54 ± 5 d before parturition (avg. temperature-humidity index = 79). Heifer calves born to these dams were either in utero heat-stressed (IU-HT; n = 36) or in utero cooled (IU-CL; n = 37) and were managed as a single cohort thereafter. A subset of heifers was euthanized at birth (d0; n = 8/treatment; 4.6 ± 2.3 h after birth) and after weaning (d63; n = 8/treatment; 63.0 ± 1.5 d) to harvest the whole MG. An ultrasound of rear mammary parenchyma (MPAR) was taken prior to d63 and correlated to harvested MPAR cross-sectional area and weight. Portions of mammary fat pad (MFP) and MPAR were preserved for compositional and histological analysis, including ductal structure number and cross-sectional area, connective tissue area, and adipocyte number and cross-sectional area. Cellular proliferation in MPAR was assessed via Ki-67 immunohistochemistry. Relative to IU-CL heifers, the MGs of IU-HT heifers were shorter in length at d0 and d63 (P ≤ 0.02). There were moderate correlations between d63 ultrasound and harvest measures. The IU-HT heifers had reduced MG and MFP mass at d0 and d63 (P ≤ 0.05), whereas MPAR mass was reduced only at d0 (P = 0.01). IU-HT heifers had greater MPAR protein and DNA content at d63 (P ≤ 0.04), but there were no MFP compositional differences (P ≥ 0.12). At d0, IU-HT heifers had fewer MPAR ductal structures (P ≤ 0.06), but there were no differences at d63. Yet, MPAR luminal and total ductal structure cross-sectional areas of IU-HT heifers were reduced at both d0 and d63 (P ≤ 0.01). The MFP adipocytes of IU-HT heifers were smaller at d0 (P ≤ 0.01), but differences were not detected at d63. The IU-HT heifers had diminished MPAR total, stromal, and epithelial cellular proliferation at both d0 and d63 (P < 0.01). Prenatal hyperthermia derails dairy calf early-life mammary development with potential carry-over consequences on future synthetic capacity.

Late-gestation in utero heat stress in dairy cattle negatively affects the mammary microstructure and milk yield at maturity, but investigation into early-life windows of mammary development is needed to fully characterize the lifelong consequences of intrauterine heat stress on the mammary gland (MG). The present study quantified mammary gross morphology and mammary fat pad and parenchyma composition, tissue microstructure, and cellular proliferation at birth and after weaning from heifers exposed to late-gestation prenatal hyperthermia. The whole MGs and fat pads of in utero heat-stressed heifers are lighter across early life relative to in utero cooled heifers. The mammary parenchyma is smaller at birth with stunted ductal development and cellular proliferation at birth and after weaning. These impairments may limit later mammary epithelial development and impact long-term productivity.

Effect of calving season on metritis incidence and bacterial content of the vagina in dairy cows.

Heat stress and uterine diseases, including metritis and endometritis, both reduce milk yields and reduce reproductive performance. Bacterial growth is promoted by elevated temperature while heat stress reduces host immune cell function, but it is not known whether increased environmental temperature promotes uterine disease by altering host immunity or bacterial growth. We hypothesize that seasonal variations in environmental temperature influence metritis incidence in the dairy cow independent of bacterial prevalence in the reproductive tract. To investigate how environmental temperature may impact metritis incidence, records of 3507 calvings in Florida over a 5-year period were evaluated. The incidence of metritis increased from 21.1% in the cool season (October through March) to 24.2% during the warm season (April through September, P < 0.05). To elucidate a link between environmental temperature and uterine disease, 102 cows were enrolled during the warm season (September 2017; n = 51) and cool season (February-March 2018; n = 51). Cows were maintained on pasture during the dry period and moved to free stall barns with fans and water soakers immediately prior to calving and remained in that environment after calving. Vaginal mucus was collected and scored on days 7 (to evaluate metritis) and 21 (to evaluate endometritis) postpartum to evaluate the incidence of uterine disease and quantify bacterial content and species using qPCR. Daily milk yield for the first 60 DIM was reduced during the warm season compared with the cool season (32.6 ± 1.62 vs 37.23 ± 1.60 kg, P < 0.05) consistent with effects of prepartum heat stress. Interestingly, more cows had persistent uterine disease on both d 7 and d 21 in the warm season compared with the cool season (58.0 vs 29.4%, P < 0.05). Regardless of calving season the total bacterial content in the vagina was greater on d 7 compared to d 21. While metritis incidence was increased in the warm season, the vaginal content of total bacteria, Escherichia coli, Trueperella pyogenes, Fusobacterium necrophorum and Prevotella melaninogenica were similar during the cool season and the warm season. Our data suggests that prepartum heat stress related to season of calving increased the incidence of metritis and persistence of uterine disease in the dairy cow independent of vaginal bacteria content. The possibility that prepartum heat stress perturbs host immune function and increases the risk of metritis when cows are exposed to an equivalent number of pathogenic bacteria requires further investigation.

Dry Period Heat Stress Impacts Mammary Protein Metabolism in the Subsequent Lactation.

Dry period heat stress impairs subsequent milk production, but its impact on milk protein content and yield is inconsistent. We hypothesize that dairy cow exposure to dry period heat stress will reduce milk protein synthesis in the next lactation, potentially through modified amino acid (AA) transport and compromised mTOR signaling in the mammary gland. Cows were enrolled into heat-stressed (dry-HT, n = 12) or cooled (dry-CL, n = 12) treatments for a 46-day dry period then cooled after calving. Milk yield and composition and dry matter intake were recorded, and milk, blood, and mammary tissue samples were collected at 14, 42, and 84 days in milk (DIM) to determine free AA concentrations, milk protein fractions, and mammary AA transporter and mTOR pathway gene and protein expression. Dry matter intake did not significantly differ between treatments pre- or postpartum. Compared with dry-CL cows, milk yield was decreased (32.3 vs. 37.7 ± 1.6 kg/day) and milk protein yield and content were reduced in dry-HT cows by 0.18 kg/day and 0.1%. Further, dry-HT cows had higher plasma concentrations of glutamic acid, phenylalanine, and taurine. Gene expression of key AA transporters was upregulated at 14 and 42 DIM in dry-HT cows. Despite minor changes in mTOR pathway gene expression, the protein 4E-BP1 was upregulated in dry-HT cows at 42 DIM whereas Akt and p70 S6K1 were downregulated. These results indicate major mammary metabolic adaptations during lactation after prior exposure to dry period heat stress.

Endocrine Signals Altered by Heat Stress Impact Dairy Cow Mammary Cellular Processes at Different Stages of the Dry Period.

Hormonal alterations occurring under late gestation heat stress may disturb mammary gland remodelling, resulting in a reduced milk yield during the subsequent lactation. We investigated the effects of an altered endocrine environment on mammary gene expression at different stages of the dry period. Mammary gland biopsies from in vivo-cooled (CL) or heat-stressed (HT) cows were collected at d 3 and 35 relative to dry-off and divided into explants. Explants were incubated in vitro for 24 h in one of three media: Basal: no prolactin or estrogen; CL-mimic: Basal + low prolactin + high 17ß-estradiol, or HT-mimic: Basal + high prolactin + low 17ß-estradiol. Real time qPCR was used to quantify gene expression. We established that late-gestation heat stress changes the expression of prolactin and oestrogen receptors, downregulates genes involved in apoptosis, autophagy and proliferation at d 3 and upregulates genes related to those cellular processes at d 35. Moreover, compared with in vivo treatments, we showed that the expression of fewer genes was impacted by in vitro treatments which aimed to mimic the hormonal response of cows exposed to a different environment. Further research will continue to uncover the mechanisms behind the production impairments caused by late-gestation heat stress.

Investigating the Use of Dry Matter Intake and Energy Balance Prepartum as Predictors of Digestive Disorders Postpartum.

One objective was to evaluate the association of dry matter intake as a percentage of body weight (DMI%BW) and energy balance (EB) prepartum and postpartum, and energy-corrected milk (ECM) postpatum with digestive disorders postpartum. For this, ANOVA was used, and DMI%BW, EB, and ECM were the outcome variables, and left displaced abomasum (LDA), indigestion, and other digestive disorders (ODDZ) were the explanatory variables. The main objective was to evaluate prepartum DMI%BW and EB as predictors of digestive disorders. For this, logistic regression was used, and LDA, indigestion, and ODDZ were the outcome variables and DMI%BW and EB were the explanatory variables. Data from 689 cows from 11 experiments were compiled. Left displaced abomasum was not associated with prepartum DMI%BW or EB. Postpartum data were normalized to the day of the event (day 0). Cows that developed LDA had lesser postpartum DMI%BW on days -24, -23, -12, -7 to 0 and from days 1 to 8, 10 to 12, and 14 and 16, lesser postpartum EB from days -7 to -5, -3 to 0, and 12, and lesser postpartum energy-corrected milk on days -19, -2, -1, 0, 7, 9, 10, 15, and 17 relative to diagnosis than cows without LDA. Cows that developed indigestion had lesser prepartum DMI%BW and EB than cows without indigestion, and lesser postpartum DMI%BW on days -24, -1, 0, 1, and 2, and greater DMI%BW on day 26, lesser ECM on days -24, -2, -1, 0, 1, and 2 relative to diagnosis. Postpartum EB was not associated with indigestion postpartum. Cows that developed ODDZ had lesser prepartum DMI%BW on day -8 and from days -5 to -2, lesser prepartum EB on day -8 and from days -5 to -2, and lesser postpartum DMI%BW than cows without ODDZ. Each 0.1 percentage point decrease in the average DMI%BW and each Mcal decrease in the average EB in the last 3 days prepartum increased the odds of having indigestion by 9% each. Cutoffs for DMI%BW and EB during the last 3 days prepartum to predict indigestion were established and were ≤1.3%/day and ≤0.68 Mcal/day, respectively. In summary, measures of prepartum DMI%BW and EB were associated with indigestion and ODDZ postpartum and were predictors of indigestion postpartum, although the effect sizes were small.

Effect of colostral volume, interval between calving and first milking, and photoperiod on colostral IgG concentrations in dairy cows.

OBJECTIVE: To identify cow and management factors associated with colostral IgG concentration in dairy cows. DESIGN: Prospective observational study. ANIMALS: 81 multiparous Holstein-Friesian cows from a single herd. PROCEDURES: Serum was obtained at the start of the nonlactating period, and cows were assigned to 1 of 4 photoperiod groups: natural day length (n = 22 cows), long days (16 h of light/d [21]) or short days (8 h of light/d [20]) for the entire nonlactating period, or natural day length followed by short days for the last 21 days of the nonlactating period (18). Serum and colostrum were collected at the first milking after calving. Regression analysis was used to investigate associations between colostral IgG concentration and the interval between calving and first milking, colostral volume, photoperiod, length of the nonlactating period, and season of calving. RESULTS: Colostral IgG concentration decreased by 3.7% during each subsequent hour after calving because of postparturient secretion by the mammary glands. The interval between calving and first milking and the colostral volume were significantly and negatively associated with colostral IgG concentration, with the former effect predominating. Photoperiod had no effect on colostral IgG concentration or volume. Serum protein concentration at calving correlated poorly with colostral IgG concentration. CONCLUSIONS AND CLINICAL RELEVANCE: Dairy producers should harvest colostrum as soon as possible after calving to optimize transfer of passive immunity in neonatal calves. Photoperiod can be manipulated without adversely affecting colostral IgG concentration.

Heat Stress Impacts Immune Status in Cows Across the Life Cycle.

Heat stress has a myriad of effects on dairy cattle across the life cycle. Whereas, the most commonly recognized impacts are associated with production responses, emerging evidence indicates that heat stress profoundly alters the immune response of calves and cows, from the prenatal stage through lactation. For example, in utero heat stress reduces passive immune transfer regardless of colostrum source, relative to normothermic conditions in late gestation. Dry cows exposed to heat stress have lower immunoglobulin responses to ovalbumin vaccination, but this effect dissipates with cooling following parturition. Conversely, cows under heat stress when dry exhibit carryover effects on the innate arm of the immune system in early lactation. In this paper we review the effects of heat stress throughout the life cycle of the dairy cow, with particular emphasis on the impact of heat stress during late gestation on the cow and the developing fetus, both before and after parturition. In addition, the impact of altered immune status under heat stress on other physiological systems, especially those supporting milk production, are considered. Finally, management interventions to prevent and reverse the effect of heat stress are presented.

In Utero Heat Stress Programs Reduced Performance and Health in Calves.

Heat stress during late gestation adversely impacts the developing calf. Calves that experience heat stress are born at a lower bodyweight and those deficits persist at least until puberty. In utero heat stress reduces passive transfer and calf survival. Late gestation heat stress programs a phenotype with lower milk yield, relative to herd mates born to cooled dams, in the first lactation and subsequent lactations.