Heat stress induces autophagy in pig ovaries during follicular development.

Hyperthermia or heat stress (HS) occurs when heat dissipation mechanisms are overwhelmed by external and internal heat production. Hyperthermia negatively affects reproduction and potentially compromises oocyte integrity and reduces developmental competence of ensuing embryos. Autophagy is the process by which cells recycle energy through the reutilization of cellular components and is activated by a variety of stressors. Study objectives were to characterize autophagy-related proteins in the ovary following cyclical HS during the follicular phase. Twelve gilts were synchronized and subjected to cyclical HS (n = 6) or thermal neutral (n = 6) conditions for 5 days during the follicular phase. Ovarian protein abundance of Beclin 1 and microtubule associated protein light chain 3 beta II were each elevated as a result of HS (P = 0.001 and 0.003, respectively). The abundance of the autophagy related (ATG)12-ATG5 complex was decreased as a result of HS (P = 0.002). Regulation of autophagy and apoptosis occurs in tight coordination, and B-cell lymphoma (BCL)2 and BCL2L1 are involved in regulating both processes. BCL2L1 protein abundance, as detected via immunofluorescence, was increased in both the oocyte (∼1.6-fold; P < 0.01) and granulosa cells of primary follicles (∼1.4-fold P < 0.05) of HS ovaries. These results suggest that ovarian autophagy induction occurs in response to HS during the follicular phase, and that HS increases anti-apoptotic signaling in oocytes and early follicles. These data contribute to the biological understanding of how HS acts as an environmental stress to affect follicular development and negatively impact reproduction.

Physiological mechanisms through which heat stress compromises reproduction in pigs.

Seasonal variations in environmental temperatures impose added stress on domestic species bred for economically important production traits. These heat-mediated stressors vary on a seasonal, daily, or spatial scale, and negatively impact behavior and reduce feed intake and growth rate, which inevitably lead to reduced herd productivity. The seasonal infertility observed in domestic swine is primarily characterized by depressed reproductive performance, which manifests as delayed puberty onset, reduced farrowing rates, and extended weaning-to-estrus intervals. Understanding the effects of heat stress at the organismal, cellular, and molecular level is a prerequisite to identifying mitigation strategies that should reduce the economic burden of compromised reproduction. In this review, we discuss the effect of heat stress on an animal's ability to maintain homeostasis in multiple systems via several hypothalamic-pituitary-end organ axes. Additionally, we discuss our understanding of epigenetic programming and how hyperthermia experienced in utero influences industry-relevant postnatal phenotypes. Further, we highlight the recent recognized mechanisms by which distant tissues and organs may molecularly communicate via extracellular vesicles, a potentially novel mechanism contributing to the heat-stress response.

The impact of in utero heat stress and nutrient restriction on progeny body composition.

We recently demonstrated that in utero heat stress (IUHS) alters future tissue accretion in pigs, but whether this is a conserved response among species, is due to the direct effects of heat stress (HS) or mediated by reduced maternal feed intake (FI) is not clear. Study objectives were to compare the quantity and rate of tissue accretion in rats exposed to differing in utero thermal environments while eliminating the confounding effect of dissimilar maternal FI. On d3 of gestation, pregnant Sprague-Dawley rats (189.0±5.9g BW) were exposed to thermoneutral (TN; 22.2±0.1°C; n=8), or HS conditions (cyclical 30 to 34°C; n=8) until d18 of gestation. A third group was pair-fed to HS dams in TN conditions (PFTN; 22.2±0.1°C; n=8) from d4 to d19 of gestation. HS increased dam rectal temperature (p=0.01; 1.3°C) compared to TN and PFTN mothers, and reduced FI (p=0.01; 33%) compared to TN ad libitum fed controls. Although litter size was similar (p=0.97; 10.9 pups/litter), pup birth weight was reduced (p=0.03; 15.4%) in HS compared to PFTN and TN dams. Two male pups per dam [n=8 in utero TN (IUTN); n=8 IUHS; n=8 in utero PFTN (IUPFTN)] were selected from four dams per treatment based on similar gestation length, and body composition was determined using dual-energy x-ray absorptiometry (DXA) on d26, d46, and d66 of postnatal life. Whole-body fat content increased (p=0.01; 11.2%), and whole-body lean tissue decreased (p=0.01; 2.6%) in IUPFTN versus IUTN and IUHS offspring. Whole-body composition was similar between IUHS and IUTN offspring. Epididymal fat pad weight increased (p=0.03; 21.6%) in IUPFTN versus IUHS offspring. In summary and in contrast to pigs, IUHS did not impact rodent body composition during this stage of growth; however, IUPFTN altered the future hierarchy of tissue accretion.

Porcine endometrial heat shock proteins are differentially influenced by pregnancy status, heat stress, and altrenogest supplementation during the peri-implantation period.

Heat stress (HS) deleteriously affects multiple components of porcine reproduction and is causal to seasonal infertility. Environment-induced hyperthermia causes a HS response (HSR) typically characterized by increased abundance of intracellular heat shock proteins (HSP). Gilts exposed to HS during the peri-implantation period have compromised embryo survival, however if (or how) HS disrupts the porcine endometrium is not understood. Study objectives were to evaluate the endometrial HSP abundance in response to HS during this period and assess the effect of oral progestin (altrenogest; ALT) supplementation. Postpubertal gilts (n = 42) were artificially inseminated during behavioral estrus (n = 28) or were kept cyclic (n = 14), and randomly assigned to thermal neutral (TN; 21 ± 1 °C) or diurnal HS (35 ± 1 °C for 12 h/31.6 ± 1 °C for 12 h) conditions from day 3 to 12 postestrus (dpe). Seven of the inseminated gilts from each thermal treatment group received ALT (15 mg/d) during this period. Using quantitative PCR, transcript abundance of HSP family A (Hsp70) member 1A (HSPA1A, P = 0.001) and member 6 (HSPA6, P < 0.001), and HSP family B (small) member 8 (HSB8, P = 0.001) were increased while HSP family D (Hsp60) member 1 (HSPD1, P = 0.01) was decreased in the endometrium of pregnant gilts compared to the cyclic gilts. Protein abundance of HSPA1A decreased (P = 0.03) in pregnant gilt endometrium due to HS, while HSP family B (small) member 1 (HSPB1) increased (P = 0.01) due to HS. Oral ALT supplementation during HS reduced the transcript abundance of HSP90α family class B member 1 (HSP90AB1, P = 0.04); but HS increased HSP90AB1 (P = 0.001), HSPA1A (P = 0.02), and HSPA6 (P = 0.04) transcript abundance irrespective of ALT. ALT supplementation decreased HSP90α family class A member 1 (HSP90AA1, P = 0.001) protein abundance, irrespective of thermal environment, whereas ALT only decreased HSPA6 (P = 0.02) protein abundance in TN gilts. These results indicate a notable shift of HSP in the porcine endometrium during the peri-implantation period in response to pregnancy status and heat stress.

Heat stress (HS) deleteriously affects multiple components of porcine reproduction and causes seasonal infertility. Environment-induced hyperthermia causes a HS response (HSR) typically characterized by increased abundance of intracellular heat shock proteins (HSP). Gilts exposed to HS during the peri-implantation period have compromised embryo survival, however if (or how) HS disrupts the porcine endometrium is not understood. Study objectives were to evaluate the endometrial HSP abundance in response to HS during this period and assess the effect of oral progestin (altrenogest; ALT) supplementation. We evaluated the abundance of HSP90, HSP70, HSP60 and HSPB in the porcine endometrium during the peri-implantation period. We demonstrate how a physiological event such as pregnancy and an environmental stressor such as HS, individually and in combination, alter the endometrial abundance of these HSP. Moreover, supplementation of pregnant gilts subjected to HS with ALT also altered the abundance of these HSP in the porcine endometrium.

Muscle Lipid Droplets: Cellular Signaling to Exercise Physiology and Beyond.

Conventionally viewed as energy storage depots, lipid droplets (LDs) play a central role in muscle lipid metabolism and intracellular signaling, as recognized by recent advances in our biological understanding. Specific subpopulations of muscle LDs, defined by location and associated proteins, are responsible for distinct biological functions. In this review, the traditional view of muscle LDs is examined, and the emerging role of LDs in intracellular signaling is highlighted. The effects of chronic and acute exercise on muscle LD metabolism and signaling is discussed. In conclusion, future directions for muscle LD research are identified. The primary focus will be on human studies, with inclusion of select animal/cellular/non-muscle studies as appropriate, to provide the underlying mechanisms driving the observed findings.

Methods for reproductive tract scoring as a tool for improving sow productivity.

Improving sow lifetime productivity is essential for maximizing farm profitability. Study objectives were to determine the accuracy for different vulva scoring methods in a commercial production system and to assess whether gilt reproductive tract scoring [evaluated by vulva width (VW)] prior to puberty could serve as useful gilt selection criteria. To accomplish this objective, 958 prepubertal replacement gilts in a commercial system were evaluated at approximately 15 wk of age. Gilt body weight (BW) was recorded in addition to 4 different methods to evaluate VW. Methods for VW assessment included digital caliper measurement (mm), visual evaluation and scoring by trained farm personnel [Farm Score (FS)], and 2 methods using scoring tools [Vulva Score Method A and B (VSA and VSB, respectively)] specifically calibrated from the VW distribution measured on gilts from previous studies. The VSA and FS methods assigned gilts to one of 3 categories (S, M, L, and 1, 2, 3, respectively) whereas VSB classified gilts vulvas using a 5-point scoring system (1 to 5). At 15 wk of age, a low proportion of variability in vulva size (27.8 ± 0.1 mm) could be explained by BW (62.2 ± 0.2 kg; R 2 = 0.05). All 3 scoring methods were effective in categorizing gilts based upon VW, as the measured VW size within methods differed by score (P < 0.01). The proportion of gilts achieving their first parity increased with score for VSA (64.7%, 73.2%, and 84.4%; P = 0.02), VSB (66.0%, 71.7%, 79.2%, 76.4%, and 84.2%; P = 0.02), and FS (67.2%, 75.0%, and 88.8%; P = 0.03), but VSA, VSB, and FS did not influence percentage of gilts achieving their second parity (P = 0.32, 0.29, and 0.30, respectively). Litter performance of gilts scored as M or L using VSA improved with an increased total born over 2 parities compared to those scored as S (23.96 vs. 26.38 pigs; P < 0.01) as well as born alive (21.13 vs. 23.05 pigs; P < 0.05). Results were similar for VSB, where scores 2 to 5 had greater total born (23.97 vs. 26.33 pigs; P < 0.01) and born alive (21.11 vs. 23.02 pigs; P < 0.05) through 2 parities compared to gilts scored 1. Using the FS method, total born pigs tended to be increased (P = 0.06) through 2 parities for gilts having a 2 or 3 vulva score compared to those scored as a 1. Collectively, assessing VW at approximately 15 wk of age may identify sows with improved productivity through 2 parities as breeding herd females.

Differentiating between the effects of heat stress and lipopolysaccharide on the porcine ovarian heat shock protein response1.

Heat stress (HS) negatively affects both human and farm-animal health and undermines efficiency in a variety of economically important agricultural variables, including reproduction. HS impairs the intestinal barrier, allowing for translocation of the resident microflora and endotoxins, such as lipopolysaccharide (LPS), from the gastrointestinal lumen into systemic circulation. While much is known about the cellular function of heat shock proteins (HSPs) in most tissues, the in vivo ovarian HSP response to stressful stimuli remains ill-defined. The purpose of this study was to compare the effects of HS or LPS on ovarian HSP expression in pigs. We hypothesized that ovarian HSPs are responsive to both HS and LPS. Altrenogest (15 mg/d) was administered per os for estrus synchronization (14 d) prior to treatment and three animal paradigms were used: (i) gilts were exposed to cyclical HS (31 ± 1.4 °C) or thermoneutral (TN; 20 ± 0.5 °C) conditions immediately following altrenogest withdrawal for 5 d during follicular development; (ii) gilts were subjected to repeated (4×/d) saline (CON) or LPS (0.1 µg/kg BW) i.v. infusion immediately following altrenogest withdrawal for 5 d; and (iii) gilts were subjected to TN (20 ± 1 °C) or cyclical HS (31 to 35 °C) conditions 2 d post estrus (dpe) until 12 dpe during the luteal phase. While no differences were detected for transcript abundances of the assessed ovarian HSP, the protein abundance of specific HSP was influenced by stressors during the follicular and luteal phases. HS during the follicular phase tended (P < 0.1) to increase ovarian protein abundance of HSP90AA1 and HSPA1A, and increased (P ≤ 0.05) HSF1, HSPD1, and HSPB1 compared with TN controls, while HS decreased HSP90AB1 (P = 0.01). Exposure to LPS increased (P < 0.05) HSP90AA1 and HSPA1A and tended (P < 0.1) to increase HSF1 and HSPB1 compared with CON gilts, while HSP90AB1 and HSPD1 were not affected by LPS. HS during the luteal phase increased (P < 0.05) abundance of HSPB1 in corpora lutea (CL), decreased (P < 0.05) CL HSP90AB1, but did not impact HSF1, HSPD1, HSP90AA1, or HSPA1A abundance. Thus, these data support that HS and LPS similarly regulate expression of specific ovarian HSP, which suggest that HS effects on the ovary are in part mediated by LPS.

Effects of dietary chromium propionate on growth performance, metabolism, and immune biomarkers in heat-stressed finishing pigs1.

Study objectives were to determine the effects of chromium (Cr) propionate (Cr propionate 0.04%; 0.5 g/kg of feed to deliver 200 parts per billion Cr/d; KemTRACE Cr, Kemin Industries, Inc., Des Moines, IA) on growth performance, metabolism, and health biomarkers in heat-stressed and nutrient-restricted pigs. Crossbred barrows (n = 96; 105 ± 1 kg BW) were enlisted in an experiment conducted in two replicates, blocked by initial BW, and randomly assigned to one of six dietary-environmental treatments: (i) thermoneutral (TN) and fed ad libitum a control diet (TNCtl), (ii) TN and fed ad libitum a Cr supplemented diet (TNCr), (iii) TN and pair-fed a control diet (PFCtl), (iv) TN and pair-fed a Cr supplemented diet (PFCr), (v) heat stress (HS) and ad libitum fed a control diet (HSCtl), or (vi) HS and ad libitum fed a Cr supplemented diet (HSCr). The study consisted of three experimental periods (P). During P0 (5 d), all pigs were housed in TN conditions (21.3 ± 0.1 °C, 56.8 ± 0.3% relative humidity [RH]) and fed the control diet ad libitum. During P1 (5 d), pigs were fed their respective dietary treatments ad libitum and kept in TN conditions. During P2 (35 d), HSCtl and HSCr-treated pigs were fed ad libitum and exposed to progressive cyclical HS conditions (27 to 31 °C, 50 ± 0.3% RH), while TNCtl, TNCr, PFCtl, and PFCr pigs remained in TN conditions and were fed ad libitum or pair-fed to their respective HSCtl and HSCr counterparts to eliminate the confounding effects of dissimilar feed intake. Overall, HS pigs had increased (P < 0.01) rectal temperature, skin temperature, and respiration rate (0.3 °C, 3.8 °C, and 32 breaths per minute, respectively) relative to TN pigs. Overall, HS decreased ADFI and ADG (20 and 21%, respectively; P < 0.01) compared with TN controls. Final BW tended to be increased in HSCr (2.7 kg, P = 0.06) compared with HSCtl pigs. Similarly, ADG tended to be increased during P2 in HSCr relative to HSCtl-treatment (0.77 vs. 0.72 kg/d; P = 0.10). There were no effects of Cr on most production parameters, but ADFI tended to be increased in Cr relative to Ctl-fed pigs (3.19 vs. 3.09 kg/d; P = 0.08). No effects of Cr supplementation were detected on circulating glucose, insulin, NEFA, cholesterol, triglycerides, or lipopolysaccharide binding protein. However, blood neutrophils were increased in HSCr (37%; P < 0.01) relative to HSCtl pigs. In summary, these results suggest Cr supplementation may benefit growth performance during HS.

Characterizing the acute heat stress response in gilts: II. Assessing repeatability and association with fertility.

Mitigating heat stress (HS) in swine production is important as it detrimentally affects multiple aspects of overall animal production efficiency. Study objectives were to determine if gilts characterized as tolerant (TOL) or susceptible (SUS) in response to HS maintain that phenotype later in life and if that phenotype influences reproductive ability during HS. Individual gilts identified as TOL (n = 50) or SUS (n = 50) from a prepubertal HS challenge were selected based on their rectal temperature (TR) during acute HS. The study consisted of 4 experimental periods (P). During P0 (2 d), all pigs were exposed to thermoneutral (TN) conditions (21.1 °C). During P1 (14 d), all gilts received Matrix (15 mg altrenogest per day) to synchronize estrus, and were maintained in TN conditions. During P2 (9 d), Matrix supplementation was terminated and gilts were subjected to diurnal HS with ambient temperatures set at 35 °C from 1000 to 2200 h and 21 °C from 2200 to 1000 h. Also during P2 gilts underwent estrus detection and artificial insemination. During P3 gilts were housed in TN conditions for 41 d at which they were sacrificed and reproductive tracts were collected. During the last 2 d of P1 and throughout the entirety of P2, TR and skin temperature (TS) were recorded. During P2, SUS had increased TR relative to TOL pigs during P2 (0.27 °C; P < 0.01). Overall, uterine wet weight, ovarian weight, corpora lutea (CL) count, and embryo survival were 5.6 ± 0.1 kg, 21.6 ± 0.3 g, 17.8 ± 0.3 CLs, and 79 ± 2%, respectively, and not influenced by prepubertal HS tolerance classification (P ≥ 0.37). Tolerant gilts had a longer return-to-estrus (6.1 vs. 5.5 d, respectively; P = 0.01) following altrenogest withdrawal and tended to have larger CL diameters (10.3 vs. 10.1 mm; P = 0.06) compared to SUS gilts. Fetal weight (25.4 vs. 23.6 g; P = 0.01) and fetal crown-rump length (74.8 vs. 72.8 mm; P < 0.01) were higher in gilts previously classified as SUS compared to those previously classified as TOL. Additionally, neither litter size nor the number of fetuses detected as a percentage of ovulations was influenced by classification. In summary, SUS gilts had a shorter return-to-estrus, increased fetus size, and tended to have smaller CL diameters compared to TOL gilts. Additionally, SUS gilts also retained their inability to maintain euthermia postpubertally relative to TOL gilts. In conclusion, there appeared to be little reproductive advantage of maintaining a lower TR during HS.

Characterization of the acute heat stress response in gilts: III. Genome-wide association studies of thermotolerance traits in pigs.

Heat stress is one of the limiting factors negatively affecting pig production, health, and fertility. Characterizing genomic regions responsible for variation in HS tolerance would be useful in identifying important genetic factor(s) regulating physiological responses to HS. In the present study, we performed genome-wide association analyses for respiration rate (RR), rectal temperature (TR), and skin temperature (TS) during HS in 214 crossbred gilts genotyped for 68,549 single nucleotide polymorphisms (SNP) using the Porcine SNP 70K BeadChip. Considering the top 0.1% smoothed phenotypic variances explained by SNP windows, we detected 26, 26, 21, and 14 genes that reside within SNPs explaining the largest proportion of variance (top 25 SNP windows) and associated with change in RR (ΔRR) from thermoneutral (TN) conditions to HS environment, as well as the change in prepubertal TR (ΔTR), change in postpubertal ΔTR, and change in TS (ΔTS), respectively. The region between 28.85 Mb and 29.10 Mb on chromosome 16 explained about 0.05% of the observed variation for ΔRR. The growth hormone receptor (GHR) gene resides in this region and is associated with the HS response. The other important candidate genes associated with ΔRR (PAIP1, NNT, and TEAD4), ΔTR (LIMS2, TTR, and TEAD4), and ΔTS (ERBB4, FKBP1B, NFATC2, and ATP9A) have reported roles in the cellular stress response. The SNP explaining the largest proportion of variance and located within and in the vicinity of genes were related to apoptosis or cellular stress and are potential candidates that underlie the physiological response to HS in pigs.

The effect of recovery from heat stress on circulating bioenergetics and inflammatory biomarkers.

Heat stress (HS) jeopardizes animal productivity and health. The intestinal barrier is sensitive to HS and heat-induced hyperpermeability plays a key role in its pathophysiology. However, the biology of recovery following HS is less understood. Thus, study objectives were to determine the temporal pattern of metabolic, inflammatory, and intestinal histological parameters during HS recovery. Female pigs (n = 32; 19.5 ± 0.5 kg BW) were sacrificed following exposure to 1 of 4 environmental treatments: 1) constant thermoneutral (TN) conditions (TNC; 24.2 ± 0.5°C), 2) no TN recovery post HS (0D), 3) 3 d of TN recovery post HS (3D), and 4) 7 d of TN recovery post HS (7D). The HS protocol was cyclical (33.6 ± 1.8 to 37.4 ± 2.1°C) and lasted for 3 d for all HS treatments. During the 3 d of HS, rectal temperature, skin temperature, and respiration rates were increased (1.3°C, 4.8°C, and 77 breaths/min, respectively; P < 0.01) and ADFI was decreased (27%; P < 0.01) compared to TNC pigs. Skin temperature tended to be decreased 0.6°C in 3D pigs during days 1-3 of recovery (P = 0.06) and was decreased 1.6 and 0.7°C during days 1-3 and 4-7 of recovery, respectively, in 7D pigs (P ≤ 0.03) compared to TNC. Relative to TNC pigs, ADFI remained 14% decreased during days 1-3 of recovery in both 3D and 7D pigs, and 17% decreased during days 4-7 in 7D pigs (P ≤ 0.01). Plasma glucose was decreased (10%; P = 0.03) for 0D and 3D relative to TNC pigs. Circulating lipopolysaccharide-binding protein was increased in 3D and 7D vs. TNC pigs (110 and 147%, respectively; P = 0.01) and tended to increase linearly with increasing recovery time (P = 0.08). Circulating tumor necrosis factor alpha was decreased (15%) in 0D pigs and increased linearly with advancing recovery time (P < 0.01). Jejunum and ileum villus height were reduced 17 and 11% in 0D vs. TNC pigs and increased linearly with progressive recovery time (P < 0.01). Jejunum and ileum mucosal surface areas were reduced 17 and 9% in 0D pigs and remained decreased in the jejunum while the ileum recovered to TNC levels by day 3 of recovery. Relative to TNC pigs, goblet cell area was similar in jejunum and colon of 0D pigs but was reduced in the ileum of 0D pigs and in jejunum, ileum, and colon of 3D and 7D relative to TNC pigs (P < 0.01). In summary, HS has deleterious effects on intestinal morphology that seem to improve with recovery time. In contrast, feed consumption remained suppressed and inflammatory biomarkers indicative of leaky gut increased following the heat load.

Effects of zinc amino acid complex on biomarkers of gut integrity and metabolism during and following heat stress or feed restriction in pigs.

Study objectives were to determine the effects of zinc (Zn) amino acid complex (Availa Zn, Zinpro Corporation, Eden Prairie, MN) on metabolism, biomarkers of leaky gut, and inflammation during and following heat stress (HS) and nutrient restriction. Crossbred gilts (n = 50; 50 ± 2 kg BW) were blocked by initial BW and randomly assigned to one of five treatments: 1) thermoneutral (TN) and ad libitum fed a control diet (TNCtl), 2) TN and pair-fed a control diet (PFCtl), 3) TN and pair-fed a Zn-supplemented diet (PFZn), 4) HS and ad libitum fed a control diet (HSCtl), and 5) HS and ad libitum fed a Zn-supplemented diet (HSZn). The study consisted of 3 experimental periods (P): during P1 (7 d), all pigs were fed their respective diets ad libitum and housed in TN conditions (20.84 ± 0.03 °C, 47.11 ± 0.42% relative humidity). During P2 (7 d), HSCtl and HSZn pigs were exposed to progressive cyclical HS conditions (27 to 30 °C, 41.9 ± 0.5% relative humidity), while TNCtl, PFCtl, and PFZn pigs remained in TN conditions and were fed ad libitum or pair-fed to their respective HSCtl and HSZn counterparts. During P3 (5 d; "recovery phase"), all pigs were housed in TN conditions and fed ad libitum. Pigs exposed to HS had overall increased rectal temperature, skin temperature, and respiration rate (0.33 °C, 3.76 °C, and 27 bpm, respectively; P < 0.01). Relative to TN controls, HS decreased ADFI and ADG (28 and 35%, respectively; P < 0.05), but these variables were unaffected by dietary treatment. Additionally, circulating insulin did not differ between HS and TN pigs (P = 0.41), but was decreased in PF relative to TN pigs (P < 0.01). During recovery, no differences were observed in rectal temperature or respiration rate across treatments, but HSZn pigs had decreased skin temperature relative to TN, PF, and HSCtl pigs (P < 0.01). During P3, no Zn effects were observed in production parameters; however, PF pigs had increased ADFI and ADG relative to TN and HS treatments (P < 0.01). During P3, circulating insulin was increased in pigs that were HS relative to TN and PF pigs (75%, P < 0.05). Interestingly, tumor necrosis factor alpha (TNFα) levels were decreased during P3 (P = 0.04) in Zn relative to Ctl-fed pigs. Circulating lipopolysaccharide-binding protein was not different among periods (P > 0.10). In summary, Zn reduced TNFα (regardless of HS), and the stimulatory effect of HS on insulin secretion is amplified during HS recovery.

Heat stress causes dysfunctional autophagy in oxidative skeletal muscle.

We have previously established that 24 h of environmental hyperthermia causes oxidative stress and have implicated mitochondria as likely contributors to this process. Given this, we hypothesized that heat stress would lead to increased autophagy/mitophagy and a reduction in mitochondrial content. To address this hypothesis pigs were housed in thermoneutral (TN; 20°C) or heat stress (35°C) conditions for 1- (HS1) or 3- (HS3) days and the red and white portions of the semitendinosus collected. We did not detect differences in glycolytic muscle. Counter to our hypothesis, upstream activation of autophagy was largely similar between groups as were markers of autophagosome nucleation and elongation. LC3A/B-I increased 1.6-fold in HS1 and HS3 compared to TN (P < 0.05), LC3A/B-II was increased 4.1-fold in HS1 and 4.8-fold in HS3 relative to TN, (P < 0.05) and the LC3A/B-II/I ratio was increased 3-fold in HS1 and HS3 compared to TN suggesting an accumulation of autophagosomes. p62 was dramatically increased in HS1 and HS3 compared to TN Heat stress decreased mitophagy markers PINK1 7.0-fold in HS1 (P < 0.05) and numerically by 2.4-fold in HS3 compared to TN and BNIP3L/NIX by 2.5-fold (P < 0.05) in HS1 and HS3. Markers of mitochondrial content were largely increased without activation of PGC-1α signaling. In total, these data suggest heat-stress-mediated suppression of activation of autophagy and autophagosomal degradation, which may enable the persistence of damaged mitochondria in muscle cells and promote a dysfunctional intracellular environment.

Heat stress increases insulin sensitivity in pigs.

Proper insulin homeostasis appears critical for adapting to and surviving a heat load. Further, heat stress (HS) induces phenotypic changes in livestock that suggest an increase in insulin action. The current study objective was to evaluate the effects of HS on whole-body insulin sensitivity. Female pigs (57 ± 4 kg body weight) were subjected to two experimental periods. During period 1, all pigs remained in thermoneutral conditions (TN; 21°C) and were fed ad libitum. During period 2, pigs were exposed to: (i) constant HS conditions (32°C) and fed ad libitum (n = 6), or (ii) TN conditions and pair-fed (PFTN; n = 6) to eliminate the confounding effects of dissimilar feed intake. A hyperinsulinemic euglycemic clamp (HEC) was conducted on d3 of both periods; and skeletal muscle and adipose tissue biopsies were collected prior to and after an insulin tolerance test (ITT) on d5 of period 2. During the HEC, insulin infusion increased circulating insulin and decreased plasma C-peptide and nonesterified fatty acids, similarly between treatments. From period 1 to 2, the rate of glucose infusion in response to the HEC remained similar in HS pigs while it decreased (36%) in PFTN controls. Prior to the ITT, HS increased (41%) skeletal muscle insulin receptor substrate-1 protein abundance, but did not affect protein kinase B or their phosphorylated forms. In adipose tissue, HS did not alter any of the basal or stimulated measured insulin signaling markers. In summary, HS increases whole-body insulin-stimulated glucose uptake.

Effects of heat stress on carbohydrate and lipid metabolism in growing pigs.

Heat stress (HS) jeopardizes human and animal health and reduces animal agriculture productivity; however, its pathophysiology is not well understood. Study objectives were to evaluate the direct effects of HS on carbohydrate and lipid metabolism. Female pigs (57 ± 5 kg body weight) were subjected to two experimental periods. During period 1, all pigs remained in thermoneutral conditions (TN; 20°C) and were ad libitum fed. During period 2, pigs were exposed to: (1) constant HS conditions (32°C) and fed ad libitum (n = 7), or (2) TN conditions and pair-fed (PFTN; n = 10) to minimize the confounding effects of dissimilar feed intake. All pigs received an intravenous glucose tolerance test (GTT) and an epinephrine challenge (EC) in period 1, and during the early and late phases of period 2. After 8 days of environmental exposure, all pigs were killed and tissue samples were collected. Despite a similar reduction in feed intake (39%), HS pigs tended to have decreased circulating nonesterified fatty acids (NEFA; 20%) and a blunted NEFA response (71%) to the EC compared to PFTN pigs. During early exposure, HS increased basal circulating C-peptide (55%) and decreased the insulinogenic index (45%) in response to the GTT. Heat-stressed pigs had a reduced T3 to T4 ratio (56%) and hepatic 5'-deiodinase activity (58%). After 8 days, HS decreased or tended to decrease the expression of genes involved in oxidative phosphorylation in liver and skeletal muscle, and ATGL in adipose tissue. In summary, HS markedly alters both lipid and carbohydrate metabolism independently of nutrient intake.

Gestational heat stress alters postnatal offspring body composition indices and metabolic parameters in pigs.

The study objectives were to test the hypothesis that heat stress (HS) during gestational development alters postnatal growth, body composition, and biological response to HS conditions in pigs. To investigate this, 14 first parity crossbred gilts were exposed to one of four environmental treatments (TNTN, TNHS, HSTN, or HSHS) during gestation. TNTN and HSHS dams were exposed to thermal neutral (TN, cyclical 18-22°C) or HS conditions (cyclical 28-34°C) during the entire gestation, respectively. Dams assigned to HSTN and TNHS treatments were heat-stressed for the first or second half of gestation, respectively. Postnatal offspring were exposed to one of two thermal environments for an acute (24 h) or chronic (five weeks) duration in either constant TN (21°C) or HS (35°C) environment. Exposure to chronic HS during their growth phase resulted in decreased longissimus dorsi cross-sectional area (LDA) in offspring from HSHS and HSTN treated dams whereas LDA was larger in offspring from dams in TNTN and TNHS conditions. Irrespective of HS during prepubertal postnatal growth, pigs from dams that experienced HS during the first half of gestation (HSHS and HSTN) had increased (13.9%) subcutaneous fat thickness compared to pigs from dams exposed to TN conditions during the first half of gestation. This metabolic repartitioning towards increased fat deposition in pigs from dams heat-stressed during the first half of gestation was accompanied by elevated blood insulin concentrations (33%; P = 0.01). Together, these results demonstrate HS during the first half of gestation altered metabolic and body composition parameters during future development and in biological responses to a subsequent HS challenge.