RESUMEN
Study objectives were to determine the effects of mitoquinol (MitoQ, a mitochondrial-targeted antioxidant) on biomarkers of metabolism and inflammation during acute heat stress (HS). Crossbred barrows [nâ =â 32; 59.0â ±â 5.6 kg body weight (BW)] were blocked by BW and randomly assigned to 1 of 4 environmental-therapeutic treatments: 1) thermoneutral (TN) control (nâ =â 8; TNCon), 2) TN and MitoQ (nâ =â 8; TNMitoQ), 3) HS control (nâ =â 8; HSCon), or 4) HS and MitoQ (nâ =â 8; HSMitoQ). Pigs were acclimated for 6 d to individual pens before study initiation. The trial consisted of two experimental periods (P). During P1 (2 d), pigs were fed ad libitum and housed in TN conditions (20.6â ±â 0.8 °C). During P2 (24 h), HSCon and HSMitoQ pigs were exposed to continuous HS (35.2â ±â 0.2 °C), while TNCon and TNMitoQ remained in TN conditions. MitoQ (40 mg/d) was orally administered twice daily (0700 and 1800 hours) during P1 and P2. Pigs exposed to HS had increased rectal temperature, skin temperature, and respiration rate (+1.5 °C, +6.8 °C, and +101 breaths per minute, respectively; Pâ <â 0.01) compared to their TN counterparts. Acute HS markedly decreased feed intake (FI; 67%; Pâ <â 0.01); however, FI tended to be increased in HSMitoQ relative to HSCon pigs (1.5 kg vs. 0.9 kg, respectively; Pâ =â 0.08). Heat-stressed pigs lost BW compared to their TN counterparts (-4.7 kg vs. +1.6 kg, respectively; Pâ <â 0.01); however, the reduction in BW was attenuated in HSMitoQ compared to HSCon pigs (-3.9 kg vs. -5.5 kg, respectively; Pâ <â 0.01). Total gastrointestinal tract weight (empty tissue and luminal contents) was decreased in HS pigs relative to their TN counterparts (6.2 kg vs. 8.6 kg, respectively; Pâ <â 0.01). Blood glucose increased in HSMitoQ relative to HSCon pigs (15%; Pâ =â 0.04). Circulating non-esterified fatty acids (NEFA) increased in HS compared to TN pigs (Pâ <â 0.01), although this difference was disproportionately influenced by elevated NEFA in HSCon relative to HSMitoQ pigs (251 µEq/L vs. 142 µEq/L; Pâ <â 0.01). Heat-stressed pigs had decreased circulating insulin relative to their TN counterparts (47%; Pâ =â 0.04); however, the insulin:FI ratio tended to increase in HS relative to TN pigs (Pâ =â 0.09). Overall, circulating leukocytes were similar across treatments (Pâ >â 0.10). Plasma C-reactive protein remained similar among treatments; however, haptoglobin increased in HS relative to TN pigs (48%; Pâ =â 0.03). In conclusion, acute HS exposure negatively altered animal performance, inflammation, and metabolism, which were partially ameliorated by MitoQ.
Heat stress (HS) compromises animal health and productivity, and this causes major economic losses in almost every livestock sector. The negative consequences of HS are thought to originate from intestinal barrier dysfunction and subsequent immune activation. The underlying causes of lost intestinal integrity during HS are likely multifactorial; however, intestinal ischemia, increased accumulation of reactive oxygen species, and the ensuing epithelial oxidative damage might be potential causes. Mitochondria-targeted antioxidants, such as mitoquinol (MitoQ), are probably more effective than traditional dietary antioxidants (i.e., selenium, vitamin E) at alleviating oxidative stress, as they localize and accumulate within the mitochondria, potentiating their antioxidant activity. Thus, the present study aimed to investigate MitoQ's role during a thermal event in growing pigs. Herein, HS increased all body temperature indices, decreased feed intake (FI), and induced substantial body weight (BW) loss. Interestingly, the reduction in FI and BW was less dramatic in pigs receiving MitoQ. Changes in circulating metabolism and the acute phase response were observed due to the HS challenge; however, contrary to our expectations, these changes were not offset by MitoQ administration. Although our results suggest a positive MitoQ effect on growth performance, future studies are needed to corroborate the replicability of this response during HS.
Asunto(s)
Ubiquinona , Animales , Ubiquinona/análogos & derivados , Ubiquinona/farmacología , Ubiquinona/administración & dosificación , Masculino , Porcinos , Compuestos Organofosforados/farmacología , Compuestos Organofosforados/administración & dosificación , Antioxidantes/farmacología , Calor/efectos adversos , Respuesta al Choque Térmico/efectos de los fármacos , Enfermedades de los Porcinos/tratamiento farmacológico , Trastornos de Estrés por Calor/veterinaria , Trastornos de Estrés por Calor/tratamiento farmacológico , Distribución Aleatoria , Temperatura Corporal/efectos de los fármacosRESUMEN
Oxidative stress contributes to heat stress (HS)-mediated alterations in skeletal muscle; however, the extent to which biological sex mediates oxidative stress during HS remains unknown. We hypothesized muscle from males would be more resistant to oxidative stress caused by HS than muscle from females. To address this, male and female pigs were housed in thermoneutral conditions (TN; 20.8 ± 1.6°C; 62.0 ± 4.7% relative humidity; n = 8/sex) or subjected to HS (39.4 ± 0.6°C; 33.7 ± 6.3% relative humidity) for 1 (HS1; n = 8/sex) or 7 days (HS7; n = 8/sex) followed by collection of the oxidative portion of the semitendinosus. Although HS increased muscle temperature, by 7 days, muscle from heat-stressed females was cooler than muscle from heat-stressed males (0.3°C; P < 0.05). Relative protein abundance of 4-hydroxynonenal (4-HNE)-modified proteins increased in HS1 females compared with TN (P = 0.05). Furthermore, malondialdehyde (MDA)-modified proteins and 8-hydroxy-2'-deoxyguanosine (8-OHdG) concentration, a DNA damage marker, was increased in HS7 females compared with TN females (P = 0.05). Enzymatic activities of catalase and superoxide dismutase (SOD) remained similar between groups; however, glutathione peroxidase (GPX) activity decreased in HS7 females compared with TN and HS1 females (P ≤ 0.03) and HS7 males (P = 0.02). Notably, HS increased skeletal muscle Ca2+ deposition (P = 0.05) and was greater in HS1 females compared with TN females (P < 0.05). Heat stress increased sarco(endo)plasmic reticulum Ca2+ ATPase (SERCA)2a protein abundance (P < 0.01); however, Ca2+ ATPase activity remained similar between groups. Overall, despite having lower muscle temperature, muscle from heat-stressed females had increased markers of oxidative stress and calcium deposition than muscle from males following identical environmental exposure.NEW & NOTEWORTHY Heat stress is a global threat to human health and agricultural production. We demonstrated that following 7 days of heat stress, skeletal muscle from females was more susceptible to oxidative stress than muscle from males in a porcine model, despite cooler muscle temperatures. The vulnerability to heat stress-induced oxidative stress in females may be driven, at least in part, by decreased antioxidant capacity and calcium dysregulation.
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Respuesta al Choque Térmico , Músculo Esquelético , Estrés Oxidativo , Animales , Femenino , Masculino , Músculo Esquelético/metabolismo , Respuesta al Choque Térmico/fisiología , Factores Sexuales , Trastornos de Estrés por Calor/metabolismo , Trastornos de Estrés por Calor/fisiopatología , Porcinos , Modelos Animales de Enfermedad , Sus scrofaRESUMEN
Study objectives were to characterize the effects of citrulline (CIT) on physiological and intestinal morphology metrics during heat stress (HS) and feed restriction. Forty crossbred gilts (30â ±â 2 kg body weight [BW]) were assigned to one of five treatments: (1) thermoneutral (TN) fed ad libitum (AL) with control (CON) supplement (TNAL; nâ =â 8), (2) TN pair-fed (PF) with CON (PF-CON; nâ =â 8), (3) TN PF with CIT (PF-CIT; nâ =â 8), (4) HS AL with CON (HS-CON; nâ =â 8), and (5) HS AL with CIT (HS-CIT; nâ =â 8). During the period (P) 1 (7 d), pigs were in TN conditions (23.6 °C) and fed AL their respective supplemental treatments. During P2 (2.5 d), HS-CON and HS-CIT pigs were fed AL and exposed to cyclical HS (33.6 to 38.3 °C), while TNAL, PF-CON, and PF-CIT remained in TN and were fed either AL or PF to their HS counterparts. Citrulline (0.13 g/kg BW) was orally administered twice daily during P1 and P2. HS increased rectal temperature (Tr), skin temperature (Ts), and respiration rate (RR) relative to TN pigs (0.8 °C, 4.7 °C, and 47 breaths/min, respectively; Pâ <â 0.01). However, HS-CIT had decreased RR (7 breaths/min, Pâ =â 0.04) and a tendency for decreased Tr (0.1 °C, Pâ =â 0.07) relative to HS-CON pigs. During P2, HS pigs had decreased feed intake (22%; Pâ <â 0.01) and a tendency for decreased average daily gain (Pâ =â 0.08) relative to TNAL pigs, and by experimental design, PF pigs followed this same pattern. Circulating lipopolysaccharide-binding protein tended to be decreased (29%; Pâ =â 0.08) in PF relative to TNAL pigs and was increased (41%; Pâ =â 0.03) in HS compared to PF pigs. Jejunum villus height was decreased in PF relative to TNAL pigs (15%; Pâ =â 0.03); however, CIT supplementation improved this metric during feed restriction (16%; Pâ =â 0.10). Jejunum mucosal surface area decreased in PF (16%; Pâ =â 0.02) and tended to decrease in HS (11%; Pâ =â 0.10) compared to TNAL pigs. Ileum villus height and mucosal surface area decreased in HS compared to TNAL pigs (10 and 14%, respectively; Pâ ≤â 0.04), but both parameters were rescued by CIT supplementation (Pâ ≤â 0.08). Intestinal myeloperoxidase and goblet cell area remained similar among treatments and intestinal segments (Pâ >â 0.24). In summary, CIT supplementation slightly improved RR and Tr during HS. Feed restriction and HS differentially affected jejunum and ileum morphology and while CIT ameliorated some of these effects, the benefit appeared dependent on intestinal section and stressor type.
Heat stress (HS) negatively affects animal health and production efficiency and is a significant economic burden to global animal agriculture. Although the mechanisms responsible for reduced animal productivity during HS are complex and multifaceted, increasing evidence points to decreased intestinal barrier function as an important mediator of this response. Furthermore, HS causes a voluntary reduction in feed intake, and feed restriction independently induces gastrointestinal hyperpermeability. Loss of intestinal barrier integrity facilitates bacteria translocation across the epithelium into local and systemic circulation, thus initiating an immune response. Dietary citrulline has been shown to support gut health by improving intestinal barrier integrity and modulating intestinal inflammation. Therefore, the current study investigated the effects of citrulline supplementation on physiological and intestinal morphology parameters in heat-stressed and feed-restricted growing pigs. Herein, citrulline supplementation reduced respiration rate and rectal temperature in pigs exposed to the thermal load. Heat stress and feed restriction compromised small intestinal morphology, and while supplementing citrulline improved some of these parameters, the effects depended on the intestinal region and stressor type. Additional research is needed to evaluate the potential effects of citrulline supplementation on gut health during HS or nutrient restriction.
Asunto(s)
Alimentación Animal , Citrulina , Suplementos Dietéticos , Animales , Citrulina/farmacología , Citrulina/administración & dosificación , Suplementos Dietéticos/análisis , Femenino , Alimentación Animal/análisis , Porcinos/fisiología , Dieta/veterinaria , Privación de Alimentos , Calor , Intestinos/efectos de los fármacos , Intestinos/anatomía & histología , Intestinos/fisiología , Temperatura Corporal/efectos de los fármacos , Respuesta al Choque Térmico/efectos de los fármacosRESUMEN
The purpose of this investigation was to establish the role biological sex plays in circulating factors following heat stress (HS). Barrows and gilts (36.8â ±â 3.7 kg body weight) were kept in either thermoneutral (TN; 20.8â ±â 1.6 °C; 62.0%â ±â 4.7% relative humidity; nâ =â 8/sex) conditions or exposed to HS (39.4â ±â 0.6 °C; 33.7%â ±â 6.3% relative humidity) for either 1 (HS1; nâ =â 8/sex) or 7 (HS7; nâ =â 8/sex) d. Circulating glucose decreased as a main effect of the environment (Pâ =â 0.03). Circulating non-esterified fatty acid (NEFA) had an environmentâ ×â sex interaction (Pâ <â 0.01) as HS1 barrows had increased NEFA compared to HS1 gilts (Pâ =â 0.01) and NEFA from HS7 gilts increased compared to HS1 gilts (Pâ =â 0.02) and HS7 barrows (Pâ =â 0.04). Cortisol, insulin, glucagon, T3, and T4 were reduced as a main effect of environment (Pâ ≤â 0.01). Creatinine was increased in HS1 and HS7 animals compared to TN (Pâ ≤â 0.01), indicative of decreased glomerular filtration rate. White blood cell populations exhibited differential patterns based on sex and time. Neutrophils and lymphocytes had an environmentâ ×â sex interaction (Pâ ≤â 0.05) as circulating neutrophils were increased in HS1 barrows compared to TN and HS7 barrows, and HS1 gilts (Pâ ≤â 0.01) and HS7 barrows had less neutrophils compared to TN barrows (Pâ =â 0.01), whereas they remained similar in gilts. In contrast, barrow lymphocyte numbers were similar between groups, but in HS7 gilts they were decreased compared to TN and HS1 gilts (Pâ ≤â 0.04). In total, these data demonstrate that HS alters a host of circulating factors and that biological sex mediates, at least in part, the physiological response to HS.
Heat stress (HS) negatively impacts efficient pork production; however, the role of biological sex is largely unknown. The objective of this study was to determine the extent to which HS differentially impacted hematological parameters in barrows and gilts. To address this, 3-mo-old barrows and gilts were exposed to ambient temperature (TN) or constant HS for 1 or 7 d. Following the experimental period, blood was collected for analysis of hormones, metabolites, immune cells, and markers of organ damage. Overall, cortisol, insulin, glucagon, T3, and T4 were reduced following HS. Furthermore, 7 d of HS decreased circulating glucose, albeit slightly. Circulating fatty acids had a sex-specific response as HS1 barrows and HS7 gilts were increased compared to their environmental counterparts, though, these changes are minor compared to those expected with a similar feed restriction. HS caused immune system activation in barrows and gilts; however, circulating levels of specific white blood cells were time- and sex-dependent. Barrows appeared more resistant to HS-mediated kidney injury acutely; however, with continued heating, markers of kidney injury were similar between barrows and gilts. In total, these data suggest biological sex regulates some, but not all, aspects of HS-mediated biological changes in pigs.
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Ácidos Grasos no Esterificados , Animales , Femenino , Masculino , Porcinos/fisiología , Ácidos Grasos no Esterificados/sangre , Calor/efectos adversos , Factores Sexuales , Glucemia , Respuesta al Choque TérmicoRESUMEN
Zearalenone (ZEN), a nonsteroidal estrogenic mycotoxin, causes endocrine disruption and porcine reproductive dysfunction. Heat stress (HS) occurs when exogenous and metabolic heat accumulation exceeds heat dissipation. Independently, HS and ZEN both compromise swine reproduction; thus, the hypothesis investigated was two-pronged: that ZEN exposure would alter the ovarian proteome and that these effects would differ in thermal neutral (TN) and HS pigs. Pre-pubertal gilts (nâ =â 38) were fed ad libitum and assigned to either (TN: 21.0â ±â 0.1 °C) or HS (12 h cyclic temperatures of 35.0â ±â 0.2 °C and 32.2â ±â 0.1 °C). Within the TN group, a subset of pigs were pair-fed (PF) to the amount of feed that the HS gilts consumed to eliminate the confounding effects of dissimilar nutrient intake. All gilts orally received a vehicle control (CT) or ZEN (40 µg/kg/BW) resulting in six treatment groups: thermoneutral (TN) vehicle control (TC; nâ =â 6); TN ZEN (TZ; nâ =â 6); PF vehicle control (PC; nâ =â 6); PF ZEN (PZ; nâ =â 6); HS vehicle control (HC; nâ =â 7); or HS ZEN (HZ; nâ =â 7) for 7 d. When compared to the TC pigs, TZ pigs had 45 increased and 39 decreased proteins (Pâ ≤â 0.05). In the HZ pigs, 47 proteins were increased and 61 were decreased (Pâ ≤â 0.05). Exposure to ZEN during TN conditions altered sec61 translocon complex (40%), rough endoplasmic reticulum membrane (8.2%), and proteasome complex (5.4%), asparagine metabolic process (0.60%), aspartate family amino acid metabolic process (0.14%), and cellular amide metabolic process (0.02%) pathways. During HS, ZEN affected cellular pathways associated with proteasome core complex alpha subunit complex (0.23%), fibrillar collagen trimer (0.14%), proteasome complex (0.05%), and spliceosomal complex (0.03%). Thus, these data identify ovarian pathways altered by ZEN exposure and suggest that the molecular targets of ZEN differ in TN and HS pigs.
Zearalenone (ZEN) is an estrogenic mycotoxin that impairs fertility in swine. This study was designed to identify the ovarian molecular impacts of ZEN exposure in thermal neutral (TN) pre-pubertal pigs. Additionally, whether heat stress (HS) would affect the ovarian ZEN response was also queried. Using a mass spectrometry approach, proteins that are altered in the ovaries of TN and HS pigs were noted to include those involved with chemical detoxification, metabolism, and inflammation. These findings may be of use in developing mitigation strategies to improve fertility in swine exposed to ZEN via contaminated feeds.
Asunto(s)
Ovario , Proteoma , Zearalenona , Animales , Zearalenona/toxicidad , Femenino , Ovario/efectos de los fármacos , Ovario/metabolismo , Proteoma/efectos de los fármacos , Porcinos , Calor/efectos adversos , Respuesta al Choque Térmico/efectos de los fármacos , Estrógenos no Esteroides/farmacologíaRESUMEN
Heat stress (HS) occurs when exogenous and metabolic heat accumulation exceeds heat dissipation; a thermal imbalance that compromises female reproduction. This study investigated the hypothesis that HS alters the ovarian proteome and negatively impacts proteins engaged with insulin signaling, inflammation, and ovarian function. Prepubertal gilts (nâ =â 19) were assigned to one of three environmental groups: thermal neutral with ad libitum feed intake (TN; nâ =â 6), thermal neutral pair-fed (PF; nâ =â 6), or HS (nâ =â 7). For 7 d, HS gilts were exposed to 12-h cyclic temperatures of 35.0â ±â 0.2 °C and 32.2â ±â 0.1 °C, while TN and PF gilts were housed at 21.0â ±â 0.1 °C. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was performed on ovarian protein homogenates. Relative to TN gilts, 178 proteins were altered (Pâ ≤â 0.05, log2foldchangeâ ≥â 1) by HS, with 76 increased and 102 decreased. STRING gene ontology classified and identified 45 biological processes including those associated with chaperone protein refolding, cytoplasmic translational initiation, and immune activation; with a protein-protein interaction web network of 158 nodes and 563 edges connected based on protein function (FDRâ ≤â 0.05). Relative to PF, HS altered 330 proteins (Pâ ≤â 0.05, log2foldchangeâ ≥â 1), with 151 increased and 179 decreased. Fifty-seven biological pathways associated with protein function and assembly, RNA processing, and metabolic processes were identified, with a protein-protein interaction network of 303 nodes and 1,606 edges. Comparing HS with both the TN and PF treatments, 72 ovarian proteins were consistently altered by HS with 68 nodes and 104 edges, with biological pathways associated with translation and gene expression. This indicates that HS alters the ovarian proteome and multiple biological pathways and systems in prepubertal gilts; changes that potentially contribute to female infertility.
Heat stress impairs female fertility, yet the mechanisms underlying reduced fecundity remain unclear. This study investigated the ovarian proteomic changes resultant from heat stress in prepubertal gilts and discovered changes related to several important biological processes that could be responsible for reduced female fertility.
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Proteoma , Espectrometría de Masas en Tándem , Porcinos , Femenino , Animales , Cromatografía Liquida/veterinaria , Espectrometría de Masas en Tándem/veterinaria , Sus scrofa , Respuesta al Choque Térmico , CalorRESUMEN
The influence of systemic immune activation on whole-body calcium (Ca) trafficking and gastrointestinal tract (GIT) physiology is not clear. Thus, the study objectives were to characterize the effects of lipopolysaccharide (LPS) on Ca pools and GIT dynamics to increase understanding of immune-induced hypocalcemia, ileus, and stomach hemorrhaging. Twelve crossbred pigs [44â ±â 3 kg body weight (BW)] were randomly assigned to 1 of 2 intramuscular treatments: (1) control (CON; 2 mL saline; nâ =â 6) or (2) LPS (40 µg LPS/kg BW; nâ =â 6). Pigs were housed in metabolism stalls to collect total urine and feces for 6 h after treatment administration, at which point they were euthanized, and various tissues, organs, fluids, and digesta were weighed, and analyzed for Ca content. Data were analyzed with the MIXED procedure in SAS 9.4. Rectal temperature and respiration rate increased in LPS relative to CON pigs (1.4 °C and 32%, respectively; Pâ ≤â 0.05). Inflammatory biomarkers such as circulating alkaline phosphatase, aspartate aminotransferase, and total bilirubin increased in LPS compared with CON pigs whereas albumin decreased (Pâ ≤â 0.02). Plasma glucose and urea nitrogen decreased and increased, respectively, after LPS (43% and 80%, respectively; Pâ <â 0.01). Pigs administered LPS had reduced circulating ionized calcium (iCa) compared to CON (15%; Pâ <â 0.01). Considering estimations of total blood volume, LPS caused an iCa deficit of 23 mg relative to CON (Pâ <â 0.01). Adipose tissue and urine from LPS pigs had reduced Ca compared to CON (39% and 77%, respectively; Pâ ≤â 0.05). There did not appear to be increased Ca efflux into GIT contents and no detectable increases in other organ or tissue Ca concentrations were identified. Thus, while LPS caused hypocalcemia, we were unable to determine where circulating Ca was trafficked. LPS administration markedly altered GIT dynamics including stomach hemorrhaging, diarrhea (increased fecal output and moisture), and reduced small intestine and fecal pH (Pâ ≤â 0.06). Taken together, changes in GIT physiology suggested dyshomeostasis and alimentary pathology. Future research is required to fully elucidate the etiology of immune activation-induced hypocalcemia and GIT pathophysiology.
Lipopolysaccharide (LPS) activates the immune system and this is accompanied with hypocalcemia and altered gastrointestinal tract (GIT) physiology. The study objectives were to characterize whole-body calcium (Ca) trafficking and evaluate GIT dynamics during LPS-induced immune activation. Ca concentrations were analyzed after intramuscular LPS injection. Administering LPS caused marked alterations in metabolic and inflammatory biomarkers and GIT dynamics, characterized by increased lower GIT motility and stomach hemorrhaging. Circulating Ca and adipose tissue and urine Ca output were decreased after LPS. Ca concentrations in other tissues and GIT contents were not detectably different. Thus, we were unable to account for about 110 mg Ca following LPS. Where and how circulating Ca is partitioned during immune activation remains unclear.
Asunto(s)
Calcio , Tracto Gastrointestinal , Lipopolisacáridos , Animales , Femenino , Masculino , Calcio/metabolismo , Tracto Gastrointestinal/efectos de los fármacos , Tracto Gastrointestinal/metabolismo , Lipopolisacáridos/farmacología , Distribución Aleatoria , Porcinos , Enfermedades de los Porcinos/inducido químicamenteRESUMEN
Objectives were to examine the temporal pattern of intestinal mast cell dynamics and the effects of a mast cell stabilizer (ketotifen [Ket]) during acute heat stress (HS) in growing pigs. Crossbred barrows (nâ =â 42; 32.3â ±â 1.9 kg body weight [BW]) were randomly assigned to 1 of 7 environmental-therapeutic treatments: (1) thermoneutral (TN) control (TNCon; nâ =â 6), (2) 2 h HS control (2 h HSCon; nâ =â 6), (3) 2 h HSâ +â Ket (2 h HSKet; nâ =â 6); (4) 6 h HSCon (nâ =â 6), (5) 6 h HSKet (nâ =â 6), (6) 12 h HSCon (nâ =â 6), or (7) 12 h HSKet (nâ =â 6). Following 5 d of acclimation to individual pens, pigs were enrolled in two experimental periods (P). During P1 (3 d), pigs were housed in TN conditions (21.5â ±â 0.8 °C) for the collection of baseline measurements. During P2, TNCon pigs remained in TN conditions for 12 h, while HS pigs were exposed to constant HS (38.1â ±â 0.2 °C) for either 2, 6, or 12 h. Pigs were euthanized at the end of P2, and blood and tissue samples were collected. Regardless of time or therapeutic treatment, pigs exposed to HS had increased rectal temperature, skin temperature, and respiration rate compared to their TNCon counterparts (1.9 °C, 6.9° C, and 119 breaths/min; Pâ <â 0.01). As expected, feed intake and BW gain markedly decreased in HS pigs relative to their TNCon counterparts (Pâ <â 0.01). Irrespective of therapeutic treatment, circulating corticotropin-releasing factor decreased from 2 to 12 h of HS relative to TNCon pigs (Pâ <â 0.01). Blood cortisol increased at 2 h of HS (2-fold; Pâ =â 0.04) and returned to baseline by 6 h. Plasma histamine (a proxy of mast cell activation) remained similar across thermal treatments and was not affected by Ket administration (Pâ >â 0.54). Independent of Ket or time, HS increased mast cell numbers in the jejunum (94%; Pâ <â 0.01); however, no effects of HS on mast cell numbers were detected in the ileum or colon. Jejunum and ileum myeloperoxidase area remained similar among treatments (Pâ >â 0.58) but it tended to increase (12%; Pâ =â 0.08) in the colon in HSCon relative to TNCon pigs. Circulating lymphocytes and basophils decreased in HSKet relative to TN and HSCon pigs (Pâ ≤â 0.06). Blood monocytes and eosinophils were reduced in HS pigs relative to their TNCon counterparts (Pâ <â 0.01). In summary, HS increased jejunum mast cell numbers and altered leukocyte dynamics and proinflammatory biomarkers. However, Ket administration had no effects on mast cell dynamics measured herein.
Heat stress (HS) affects various physiological, metabolic, and endocrine parameters, ostensibly due to reduced intestinal barrier integrity and the ensuing immune response. Evidence indicates that generalized "stress" may be a critical component of HS-induced leaky gut, a mechanism likely mediated by mast cells. Mast cell activation has been extensively associated with various stress-related intestinal inflammatory conditions; however, its contribution to intestinal barrier dysfunction during HS remains unclear. Thus, this study was designed to evaluate mast cell dynamics during an acute HS challenge and to assess the effects a mast cell stabilizer on biomarkers of intestinal inflammation. Herein, HS induced a rapid increase in circulating cortisol, increased jejunum mast cell numbers, and altered metabolism, leukocyte dynamics, and proinflammatory biomarkers. Contrary to our hypothesis, HS did not alter circulating histamine (a biomarker of mast cell activation), and mast cell stabilization did not affect mast cell numbers nor altered histamine concentrations. Altogether, our observations support a connection between HS and intestinal mast cell infiltration that may contribute to the pathophysiology of intestinal dysfunction during a heat load.
Asunto(s)
Trastornos de Estrés por Calor , Enfermedades de los Porcinos , Porcinos , Animales , Dieta , Mastocitos , Respuesta al Choque Térmico , Temperatura Cutánea , Recto , Calor , Trastornos de Estrés por Calor/veterinariaRESUMEN
Independently, both heat stress (HS) and zearalenone (ZEN) compromise female reproduction, thus the hypothesis that ZEN would affect phenotypic, endocrine, and metabolic parameters in pigs with a synergistic and/or additive impact of HS was investigated. Prepubertal gilts (n = 6-7) were assigned to: thermoneutral (TN) vehicle control (TC; n = 6); TN ZEN (40 µg/kg; TZ; n = 6); pair-fed (PF; n = 6) vehicle control (PC; n = 6); PF ZEN (40 µg/kg; PZ; n = 6); HS vehicle control (HC; n = 7); and HS ZEN (40 µg/kg; HZ; n = 7) and experienced either constant 21.0 ± 0.10 °C (TN and PF) or 35.0 ± 0.2 °C (12 h) and 32.2 ± 0.1 °C (12 h) to induce HS for 7 d. Elevated rectal temperature (P < 0.01) and respiration rate (P < 0.01) confirmed induction of HS. Rectal temperature was decreased (P = 0.03) by ZEN. Heat stress decreased (P < 0.01) feed intake, body weight, and average daily gain, with absence of a ZEN effect (P > 0.22). White blood cells, hematocrit, and lymphocytes decreased (P < 0.04) with HS. Prolactin increased (P < 0.01) in PC and PZ and increased in HZ females (P < 0.01). 17ß-estradiol reduced (P < 0.01) in HC and increased in TZ females (P = 0.03). Serum metabolites were altered by both HS and ZEN. Neither HS nor ZEN impacted ovary weight, uterus weight, teat size or vulva area in TN and PF treatments, although ZEN increased vulva area (P = 0.02) in HS females. Thus, ZEN and HS, independently and additively, altered blood composition, impacted the serum endocrine and metabolic profile and increased vulva size in prepubertal females, potentially contributing to infertility.
Asunto(s)
Zearalenona , Porcinos , Femenino , Animales , Zearalenona/toxicidad , Sus scrofa , Respuesta al Choque Térmico , Ingestión de Alimentos , Frecuencia Respiratoria , CalorRESUMEN
Prolonged exposure to heat can lead to environment-induced heat stress (EIHS), which may jeopardize human health, but the extent to which EIHS affects cardiac architecture and myocardial cell health are unknown. We hypothesized EIHS would alter cardiac structure and cause cellular dysfunction. To test this hypothesis, 3-mo old female pigs were exposed to thermoneutral (TN; 20.6 ± 0.2 °C; n = 8) or EIHS (37.4 ± 0.2 °C; n = 8) conditions for 24 h, hearts were removed and dimensions measured, and portions of the left ventricle (LV) and right ventricle (RV) were collected. Environment-induced heat stress increased rectal temperature 1.3 °C (P < 0.01), skin temperature 11 °C (P < 0.01) and respiratory rate 72 breaths per minute (P < 0.01). Heart weight and length (apex to base) were decreased by 7.6% (P = 0.04) and 8.5% (P = 0.01), respectively, by EIHS, but heart width was similar between groups. Left ventricle wall thickness was increased (22%; P = 0.02) and water content was decreased (8.6%; P < 0.01) whereas in RV, wall thickness was decreased (26%; P = 0.04) and water content was similar in EIHS compared to TN. We also discovered ventricle-specific biochemical changes such that in RV EIHS increased heat shock proteins, decreased AMPK and AKT signaling, decreased activation of mTOR (35%; P < 0.05), and increased expression of proteins that participate in autophagy. In LV, heat shock proteins, AMPK and AKT signaling, activation of mTOR, and autophagy-related proteins were largely similar between groups. Biomarkers suggest EIHS-mediated reductions in kidney function. These data demonstrate EIHS causes ventricular-dependent changes and may undermine cardiac health, energy homeostasis, and function.
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Proteínas Quinasas Activadas por AMP , Trastornos de Estrés por Calor , Animales , Femenino , Humanos , Trastornos de Estrés por Calor/veterinaria , Proteínas de Choque Térmico , Respuesta al Choque Térmico , Proteínas Proto-Oncogénicas c-akt , Porcinos , Serina-Treonina Quinasas TOR , Ventrículos Cardíacos/fisiopatologíaRESUMEN
INTRODUCTION: The effects of lipopolysaccharides (i.e., endotoxin; LPS) on metabolism are poorly defined in lactating dairy cattle experiencing hyperlipidemia. OBJECTIVES: Our objective was to explore the effects of acute intravenous LPS administration on metabolism in late-lactation Holstein cows experiencing hyperlipidemia induced by intravenous triglyceride infusion and feed restriction. METHODS: Ten non-pregnant lactating Holstein cows (273 ± 35 d in milk) were administered a single bolus of saline (3 mL of saline; n [Formula: see text] 5) or LPS (0.375 [Formula: see text]g of LPS/kg of body weight; n [Formula: see text] 5). Simultaneously, cows were intravenously infused a triglyceride emulsion and feed restricted for 16 h to induce hyperlipidemia in an attempt to model the periparturient period. Blood was sampled at routine intervals. Changes in circulating total fatty acid concentrations and inflammatory parameters were measured. Plasma samples were analyzed using untargeted lipidomics and metabolomics. RESULTS: Endotoxin increased circulating serum amyloid A, LPS-binding protein, and cortisol concentrations. Endotoxin administration decreased plasma lysophosphatidylcholine (LPC) concentrations and increased select plasma ceramide concentrations. These outcomes suggest modulation of the immune response and insulin action. Lipopolysaccharide decreased the ratio of phosphatidylcholine to phosphatidylethanomanine, which potentially indicate a decrease in the hepatic activation of phosphatidylethanolamine N-methyltransferase and triglyceride export. Endotoxin administration also increased plasma concentrations of pyruvic and lactic acids, and decreased plasma citric acid concentrations, which implicate the upregulation of glycolysis and downregulation of the citric acid cycle (i.e., the Warburg effect), potentially in leukocytes. CONCLUSION: Acute intravenous LPS administration decreased circulating LPC concentrations, modified ceramide and glycerophospholipid concentrations, and influenced intermediary metabolism in dairy cows experiencing hyperlipidemia.
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Hiperlipidemias , Insulinas , Animales , Bovinos , Ceramidas , Ácido Cítrico , Emulsiones/farmacología , Endotoxinas/farmacología , Ácidos Grasos , Femenino , Glicerofosfolípidos , Hidrocortisona/farmacología , Hiperlipidemias/inducido químicamente , Insulinas/farmacología , Lactancia , Lipidómica , Lipopolisacáridos/farmacología , Lisofosfatidilcolinas/farmacología , Metaboloma , Metabolómica , Fosfatidilcolinas , Fosfatidiletanolamina N-Metiltransferasa/farmacología , Proteína Amiloide A Sérica , TriglicéridosRESUMEN
Heat stress (HS) and Zearalenone (ZEN) exposure affect growth, production efficiency, and animal welfare; and, under extreme situations, both can be lethal. Given that both HS and ZEN independently cause oxidative stress, we hypothesized that simultaneous exposure to HS and ZEN would cause greater oxidative stress in porcine skeletal muscle than either condition, alone. To address this hypothesis, crossbred, prepubertal gilts were treated with either vehicle control (cookie dough) or ZEN (40 µg/kg) and exposed to either thermoneutral (TN; 21.0 °C) or 12-h diurnal HS conditions (night: 32.2 °C; day: 35.0 °C) for 7 d. Pigs were euthanized immediately following the environmental challenge and the glycolytic (STW) and oxidative (STR) portions of the semitendinosus muscle were collected for analysis. In STR, malondialdehyde (MDA) concentration, a marker of oxidative stress, tended to increase following ZEN exposure (P = 0.08). HS increased CAT (P = 0.019) and SOD1 (P = 0.049) protein abundance, while ZEN decreased GPX1 protein abundance (P = 0.064) and activity (P = 0.036). In STR, HS did not alter protein expression of HSP27, HSP70, or HSP90. Conversely, in STW, MDA-modified proteins remained similar between all groups. Consistent with STR, ZEN decreased GPX1 (P = 0.046) protein abundance in STW. In STW, ZEN decreased protein abundance of HSP27 (P = 0.032) and pHSP27 (P = 0.0068), while HS increased protein expression of HSP70 (P = 0.04) and HSP90 (P = 0.041). These data suggest a muscle fiber type-specific response to HS or ZEN exposure, potentially rendering STR more susceptible to HS- and/or ZEN-induced oxidative stress, however, the combination of HS and ZEN did not augment oxidative stress.
Heat stress (HS) and Zearalenone (ZEN), a toxic feed contaminant, affect growth, production efficiency, and animal welfare, and can cause death. As HS and ZEN independently increase oxidative stress, an imbalance of free radical production and clearance, and the likelihood of ZEN contamination during heat events, we hypothesized concomitant exposure would induce oxidative stress in pig skeletal muscle more than either agent alone. To address this, female pigs were treated with a placebo or low dose of ZEN and exposed to ambient temperature or a mild cyclic HS designed to mimic environmental conditions (hot days, cooler nights) for 7 d. Following these treatments, fast- and slow-twitch muscles were collected for analysis. In slow-twitch muscle, we observed increased markers of oxidative stress in pigs exposed to ZEN primarily driven by HS and ZEN treated pigs. Additionally, ZEN reduced antioxidant abundance and enzymatic activity regardless of the environment. Conversely, HS and/or ZEN did not cause oxidative stress in fast-twitch muscle, although ZEN altered antioxidant abundance. Although a mild HS and ZEN dose was used, oxidative stress markers were altered, suggesting that slow-twitch muscle is susceptible to HS- and ZEN-mediated changes. These data raise the possibility that more severe HS exposures and higher ZEN doses may compromise muscle health.
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Trastornos de Estrés por Calor , Enfermedades de los Porcinos , Zearalenona , Animales , Femenino , Proteínas de Choque Térmico HSP27/metabolismo , Proteínas HSP70 de Choque Térmico/metabolismo , Trastornos de Estrés por Calor/metabolismo , Trastornos de Estrés por Calor/veterinaria , Respuesta al Choque Térmico , Calor , Músculo Esquelético/metabolismo , Sus scrofa , Porcinos , Enfermedades de los Porcinos/metabolismo , Zearalenona/toxicidadRESUMEN
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.
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Proteínas de Choque Térmico , Respuesta al Choque Térmico , Animales , Suplementos Dietéticos , Endometrio/metabolismo , Femenino , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Respuesta al Choque Térmico/fisiología , Embarazo , Sus scrofa/metabolismo , Porcinos , Acetato de Trembolona/análogos & derivadosRESUMEN
Heat stress (HS) compromises almost every aspect of animal agriculture including reproduction. In pigs, this infecundity is referred to as seasonal infertility (SI), a phenotype including ovarian dysfunction. In multiple species, HS-induced hyperprolactinemia has been described; hence, our study objectives were to characterize and compare HS effects on circulating prolactin (PRL) and ovarian Janus kinase/signal transducer and activator of transcription (JAK-STAT) signaling during the follicular (FOL) or luteal (LUT) phases of the estrous cycle in postpubertal gilts. Gilts were estrus synchronized using altrenogest and environmental treatments began immediately after altrenogest withdrawal. For the FOL study: postpubertal gilts were allocated to constant thermoneutral (TN; n = 6; 20 ± 1.2 °C) or cyclical HS (n = 6; 25 to 32 ± 1.2 °C) conditions for 5 d. In the LUT study: postpubertal gilts were assigned to either TN (n = 7; 20 ± 2.6 °C) or cyclical HS (n = 7; 32 to 35 ± 2.6 °C) conditions from 2 to 12 days postestrus (dpe). Blood was collected by jugular venipuncture for PRL quantification on day 5 in the FOL and on day 0 and day 12 in the LUT gilts. Ovaries and corpora lutea (CL) were obtained from euthanized FOL and LUT gilts on day 5 and day 12, respectively. Western blotting was performed to quantify prolactin receptor (PRLR) and JAK/STAT pathway protein abundance. In the FOL phase, no difference (P = 0.20) in circulating PRL between thermal groups was observed. There was no effect (P ≥ 0.34) of HS on PRLR, signal transducer and activator of transcription 3 (STAT3), signal transducer and activator of transcription 5α (STAT5α), and phosphorylated signal transducer and activator of transcription α/ß tyrosine 694/699 (pSTAT5α/ßTyr694/699) abundance and Janus kinase 2 (JAK2), phosphorylated janus kinase 2 tyrosine 1007/1008 (pJAK2Tyr1007/1008), STAT1, phosphorylated signal transducer and activator of transcription 1 tyrosine 701 (pSTAT1Tyr701), phosphorylated signal transducer and activator of transcription 1 serine 727 (pSTAT1Ser727), and phosphorylated signal transducer and activator of transcription 3 tyrosine 705 (pSTAT3Tyr705) were undetectable in FOL gilt ovaries. Ovarian pSTAT5α/ßTyr694/699 abundance tended to moderately increase (4%; P = 0.07) in FOL gilts by HS. In the LUT phase, circulating PRL increased progressively from 2 to 12 dpe, but no thermal treatment-induced difference (P = 0.37) was noted. There was no effect (P ≥ 0.16) of HS on CL abundance of PRLR, pJAK2Tyr1007/1008, JAK2, STAT1, pSTAT1Tyr701, pSTAT1Ser727, pSTAT3Tyr705, STAT5α, or pSTAT5α/ßTyr694/699. In LUT phase, CL STAT3 abundance was increased (11%; P < 0.03) by HS. There was no impact of HS (P ≥ 0.76) on levels of pJAK2Tyr1007/1008 and pSTAT5α/ßTyr694/699 in LUT gilts; however, the CL pSTAT3Tyr705:STAT3 ratio tended to be decreased (P = 0.10) due to HS. These results indicate an HS-induced estrous cycle-stage-dependent effect on the ovarian JAK/STAT pathway, establishing a potential role for this signaling pathway as a potential contributor to SI.
Heat stress (HS) negatively affects reproduction in pigs, though the precise mechanisms are not understood. This study determined if HS impacts the JAK-STAT signaling pathway in the ovary during two stages of the estrous cycle: follicular and luteal. While circulating prolactin hormone level was unchanged, there were changes to some aspects of ovarian JAK-STAT signaling that could be involved in infertility induced in pigs during HS.
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Trastornos de Estrés por Calor , Enfermedades de los Porcinos , Animales , Femenino , Trastornos de Estrés por Calor/metabolismo , Trastornos de Estrés por Calor/veterinaria , Respuesta al Choque Térmico , Janus Quinasa 2/metabolismo , Janus Quinasa 2/farmacología , Quinasas Janus/metabolismo , Quinasas Janus/farmacología , Ovario/metabolismo , Prolactina/metabolismo , Receptores de Prolactina/genética , Factores de Transcripción STAT/metabolismo , Factores de Transcripción STAT/farmacología , Factor de Transcripción STAT1/metabolismo , Factor de Transcripción STAT1/farmacología , Factor de Transcripción STAT3/metabolismo , Factor de Transcripción STAT3/farmacología , Transducción de Señal , Porcinos , Enfermedades de los Porcinos/metabolismo , Tirosina/metabolismoRESUMEN
Heat stress (HS) mitigation strategies are critically needed to combat the substantial economic effects on animal agriculture. The manifestations of seasonal infertility include delayed puberty onset, reduced conception rates, decreased litter size, and increased wean to estrus interval. To assess the effects of HS during early gestation and evaluate the benefit of supplemental altrenogest (ALT) as a mitigation strategy, 30 crossbred postpubertal gilts (157â ±â 11 kg body weight) were subjected to estrous synchronization via 14 d oral administration of ALT. Artificial insemination during estrus was performed, and gilts were then placed into one of four treatment groups: HS (35â ±â 1 °C for 12 h/31.60â ±â 1 °C for 12 h) with (HSALT, nâ =â 7) or without (HSCON, nâ =â 7) 15 mg/d ALT supplementation or thermal neutral (TN; 20â ±â 1 °C) conditions with (TNALT, nâ =â 8) or without (TNCON, nâ =â 8) 15 mg/d ALT supplementation until 12 d post-estrus (dpe). Administrating ALT occurred at 0600 hours from 3 to 12 dpe, and rectal temperatures (TR) and respiration rates (RR) were recorded. Blood was collected via jugular venipuncture on 0, 4, 8, and 12 dpe. Gilts were euthanized humanely at 12 dpe followed by the collection of ovarian tissue, and uterine flushing for conceptus collection. In HS compared with TN gilts, RR and TR were increased (Pâ <â 0.01) but unaffected by ALT supplementation. Feed intake was reduced (Pâ <â 0.01) by HS but unaltered by the ALT treatment. Corpora lutea (CL) weight was reduced (Pâ <â 0.01) in HSCON gilts when compared with TNCON and HSALT gilts despite progesterone concentrations in serum and luteal tissue not being affected by treatment (Pâ ≥â 0.10). CL diameter was reduced (Pâ ≤â 0.05) in HSALT gilts compared with other treatments. Interleukin-1ß (IL1B) uterine flush concentration was not affected (Pâ >â 0.20) by environment or ALT supplementation, although moderate (Pâ =â 0.06) interaction between environment and ALT existed, as IL1B concentration in TNALT was increased (Pâ =â 0.03) compared with TNCON gilts. While environment did not affect conceptus development (Pâ =â 0.90), ALT supplementation advanced conceptus elongation (Pâ <â 0.01). Collectively, these data demonstrate that HS may affect luteal development before pregnancy establishment, and ALT increases conceptus elongation by 12 dpe.
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Estro , Acetato de Trembolona , Animales , Femenino , Tamaño de la Camada , Embarazo , Porcinos , Temperatura , Acetato de Trembolona/análogos & derivados , Acetato de Trembolona/farmacologíaRESUMEN
The effects of selenium (Se) yeast supplementation on performance, blood biochemical and antioxidant parameters, and milk Se content and speciation were evaluated. Thirty-six mid-lactation Holstein dairy cows were randomly assigned to 1 of 3 treatments: 1) control (basal diet containing Se at 0.11 mg/kg DM), 2) basal diet + 0.5 mg supplemental Se/kg DM (SY-0.5), and 3) basal diet + 5 mg supplemental Se/kg DM (SY-5). Selenium was supplemented as Se yeast. The trial consisted of a 1-week pretrial period and an 8-week experimental period. Milk somatic cell score decreased with SY-5 supplementation (P < 0.05), but other performance parameters were not affected (P > 0.05). The serum Se concentration increased with the increasing levels of Se yeast supplementation (P < 0.05), however, blood biochemical parameters showed few treatment effects. The antioxidant capacity of dairy cows was improved with Se yeast supplementation reflected in increased serum glutathione peroxidase activity (P < 0.05) and total antioxidant capacity (P = 0.08), and decreased malondialdehyde concentration (P < 0.05). Milk total Se concentration increased with Se dose (P < 0.05). Also, the selenomethionine concentration increased with Se dose from 13.0 ± 0.7 µg/kg in control to 33.1 ± 2.1 µg/kg in SY-0.5 and 530.4 ± 17.5 µg/kg in SY-5 cows (P < 0.05). Similarly, selenocystine concentration increased from 15.6 ± 0.9 µg/kg in control and 18.9 ± 1.1 µg/kg in SY-0.5 to 22.2 ± 1.5 µg/kg in SY-5 cows (P < 0.05). In conclusion, Se yeast is a good organic Se source to produce Se-enriched cow milk with increased Se species including selenomethionine and selenocystine. The results can provide useful information on milk Se species when a high dose Se yeast was supplemented in the cow diet.
RESUMEN
Multiparous, lactating Holstein cows (n = 32) were randomly assigned to one of two dietary treatments [TMR with rumen-protected Met (RPM) or TMR without RPM (CON)], and within each dietary treatment group cows were randomly assigned to one of two environmental treatment groups in a split-plot crossover design. In phase 1 (9 d), all cows were fed ad libitum and in thermoneutral conditions (TN). In phase 2 (9 d), group 1 (n = 16) was exposed to a heat stress (HS) challenge (HSC). Group 2 cows (n = 16) were pair-fed (PFTN) to HSC counterparts and remained in TN. After a 21-d washout period, the study was repeated (period 2) and the environmental treatments were inverted relative to treatments from phase 2 of period 1, while dietary treatments remained the same for each cow. During phase 1, cows in RPM had greater plasma Met concentration compared with cows in CON (59 and 30 µM, respectively; P < 0.001). Cows in PFTN had a greater decrease (P < 0.05) in plasma insulin than cows in HSC at 4 h (-2.7 µIU/mL vs. -0.7 µIU/mL) and 8 h (-7.7 µIU/mL vs. -0.4 µIU/mL) during phase 2. Compared with cows in PFTN, cows in HSC had an increase (P < 0.05) in plasma serum amyloid A (-59 µg/mL vs. +58 µg/mL), serum haptoglobin (-3 µg/mL vs. +33 µg/mL), plasma lipopolysaccharide binding protein (-0.27 and +0.11 µg/mL), and plasma interleukin-1ß (-1.9 and +3.9 pg/mL) during phase 2. In conclusion, HSC elicited immunometabolic alterations; however, there were limited effects of RPM on cows in HSC.
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Enfermedades de los Bovinos , Trastornos de Estrés por Calor , Animales , Bovinos , Dieta/veterinaria , Suplementos Dietéticos , Femenino , Trastornos de Estrés por Calor/veterinaria , Respuesta al Choque Térmico , Lactancia , Metionina , Leche , RumenRESUMEN
Porcine pregnancy establishment and maintenance are dependent on the formation of functional corpora lutea (CL). Manganese (Mn) is critical for CL function as it is a cofactor for Mn superoxide dismutase and enzymes involved in cholesterol synthesis. Previously, we have shown that luteal Mn content increased and luteal progesterone (P4) concentration decreased in the CL of gilts fed diets supplemented with an Mn-amino acid complex (Availa-Mn; Zinpro Corporation) compared with controls fed Mn sulfate. Importantly, serum P4 increased from 0 (estrus onset) to 12 d post estrus (dpe), as expected, but P4 abundance in circulation was not affected by dietary Mn source (P = 0.15). We hypothesized that a more bioavailable Mn source (which results in increased luteal Mn content) would alter the luteal proteome and abundance of mRNA associated with steroid biogenesis during the mid-luteal phase of the estrous cycle. Postpubertal gilts (n = 32) were assigned to one of the four gestation diets. The control diet (CON) contained 20 ppm of supplemental Mn in the form of Mn sulfate. Three additional diets included 20 (TRT1), 40 (TRT2), or 60 (TRT3) ppm of supplemental Mn in the form of a Mn-amino acid complex instead of Mn sulfate. Dietary treatment began at estrus synchronization (approximately 20 d before estrus) and continued through 12 dpe when gilts were euthanized and tissues were collected. Protein and total RNA extracts from the CL were used for proteomic analysis via label-free liquid chromatography with tandem mass spectrometry to assess global protein abundance and quantitative real-time polymerase chain reaction (qRT-PCR) to assess specific mRNA abundance, respectively. Compared with CON, 188, 382, and 401 proteins were differentially abundant (P < 0.10) in TRT1, TRT2, and TRT3, respectively. Gene Ontology enrichment software revealed that proteins involved in P4 signaling and cholesterol synthesis were downregulated in CL of gilts fed Mn-amino acid complex compared with controls. Quantitative RT-PCR showed that relative transcript abundance of genes encoding steroidogenic enzymes (CYP11A1 and StAR) in CL tissue was decreased in gilts from TRT2 compared with CON (P = 0.02), but TRT1 and TRT3 were not affected (P ≥ 0.30). Collectively, these data support our hypothesis that a more bioavailable dietary Mn source may influence luteal function by altering the abundance of protein and mRNA involved in steroidogenesis.
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Manganeso , Proteómica , Aminoácidos , Animales , Cuerpo Lúteo , Suplementos Dietéticos , Femenino , Embarazo , Progesterona , PorcinosRESUMEN
BACKGROUND: Heat stress (HS) occurs when body heat accumulation exceeds heat dissipation and is associated with swine seasonal infertility. HS contributes to compromised oocyte integrity and reduced embryo development. Autophagy is a potential mechanism for the oocyte to mitigate the detrimental effects of HS by recycling damaged cellular components. METHODS: To characterize the effect of HS on autophagy in oocyte maturation, we utilized an in vitro maturation (IVM) system where oocytes underwent thermal neutral (TN) conditions throughout the entire maturation period (TN/TN), HS conditions during the first half of IVM (HS/TN), or HS conditions during the second half of IVM (TN/HS). RESULTS: To determine the effect of HS on autophagy induction within the oocyte, we compared the relative abundance and localization of autophagy-related proteins. Heat stress treatment affected the abundance of two well described markers of autophagy induction: autophagy related gene 12 (ATG12) in complex with ATG5 and the cleaved form of microtubule-associated protein 1 light chain 3 beta (LC3B-II). The HS/TN IVM treatment increased the abundance of the ATG12-ATG5 complex and exacerbated the loss of LC3B-II in oocytes. The B-cell lymphoma 2 like 1 protein (BCL2L1) can inhibit autophagy or apoptosis through its interaction with either beclin1 (BECN1) or BCL2 associated X, apoptosis regulator (BAX), respectively. We detected colocalization of BCL2L1 with BAX but not BCL2L1 with BECN1, suggesting that apoptosis is inhibited under the HS/TN treatment but not autophagy. Interestingly, low doses of the autophagy inducer, rapamycin, increased oocyte maturation. CONCLUSIONS: Our results here suggest that HS increases autophagy induction in the oocyte during IVM, and that artificial induction of autophagy increases the maturation rate of oocytes during IVM. These data support autophagy as a potential mechanism activated in the oocyte during HS to recycle damaged cellular components and maintain developmental competence.
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Autofagia/fisiología , Respuesta al Choque Térmico/fisiología , Oocitos/fisiología , Oogénesis/fisiología , Animales , Autofagia/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Femenino , Respuesta al Choque Térmico/efectos de los fármacos , Inmunosupresores/farmacología , Oocitos/efectos de los fármacos , Oogénesis/efectos de los fármacos , Sirolimus/farmacología , PorcinosRESUMEN
Study objectives were to determine the effects of dietary live yeast (Saccharomyces cerevisiae strain CNCM I-4407; ActisafHR+; 0.25g/kg of feed; Phileo by Lesaffre, Milwaukee, WI) on growth performance and biomarkers of metabolism and inflammation in heat-stressed and nutrient-restricted pigs. Crossbred barrows (n = 96; 79 ± 1 kg body weight [BW]) were blocked by initial BW and randomly assigned to one of six dietary-environmental treatments: 1) thermoneutral (TN) and fed ad libitum the control diet (TNCon), 2) TN and fed ad libitum a yeast containing diet (TNYeast), 3) TN and pair-fed (PF) the control diet (PFCon), 4) TN and PF the yeast containing diet (PFYeast), 5) heat stress (HS) and fed ad libitum the control diet (HSCon), or 6) HS and fed ad libitum the yeast diet (HSYeast). Following 5 d of acclimation to individual pens, pigs were enrolled in two experimental periods (P). During P1 (7 d), pigs were housed in TN conditions (20 °C) and fed their respective dietary treatments ad libitum. During P2 (28 d), HSCon and HSYeast pigs were fed ad libitum and exposed to progressive cyclical HS (28-33 °C) while TN and PF pigs remained in TN conditions and were fed ad libitum or PF to their HSCon and HSYeast counterparts. Pigs exposed to HS had an overall increase in rectal temperature, skin temperature, and respiration rate compared to TN pigs (0.3 °C, 5.5 °C, and 23 breaths per minute, respectively; P < 0.01). During P2, average daily feed intake (ADFI) decreased in HS compared to TN pigs (30%; P < 0.01). Average daily gain and final BW decreased in HS relative to TN pigs (P < 0.01); however, no differences in feed efficiency (G:F) were observed between HS and TN treatments (P > 0.16). A tendency for decreased ADFI and increased G:F was observed in TNYeast relative to TNCon pigs (P < 0.10). Circulating insulin was similar between HS and TN pigs (P > 0.42). Triiodothyronine and thyroxine levels decreased in HS compared to TN treatments (~19% and 20%, respectively; P < 0.05). Plasma tumor necrosis factor-alpha (TNF-α) did not differ across treatments (P > 0.57) but tended to decrease in HSYeast relative to HSCon pigs (P = 0.09). In summary, dietary live yeast did not affect body temperature indices or growth performance and had minimal effects on biomarkers of metabolism; however, it tended to improve G:F under TN conditions and tended to reduce the proinflammatory mediator TNF-α during HS. Further research on the potential role of dietary live yeast in pigs during HS or nutrient restriction scenarios is warranted.