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Endoplasmic reticulum stress (ERS) is closely related to the occurrence and progression of metabolic liver disease. The treatment targeting glucose-regulated protein 94 (GRP94) for liver disease has gotten much attention, but the specific effect of GRP94 on hepatocyte apoptosis is still unclear. So far, all the studies on GRP94 have been conducted in mice or rats, and little study has been reported on pigs, which share more similarities with humans. In this study, we used low-dose (LD) and high-dose (HD) tunicamycin (TM) to establish ERS models on piglet livers and immortalized porcine hepatic stellate cells (HSCs). On the piglet ERS model we found that ERS could significantly (p < 0.01) stimulate the secretion and synthesis of insulin-like growth factor (IGF-1), IGF-1 receptor (IGF-1R), and IGF-binding protein (IGFBP)-1 and IGFBP-3; however, with the increase in ERS degree, the effect of promoting secretion and synthesis significantly (p < 0.01) decreased. In addition, the ubiquitin protein and ubiquitination-related gene were significantly increased (p < 0.05) in the LD group compared with the vehicle group. The protein level of Active-caspase 3 was significantly increased (p < 0.01) in the HD group, however, the TUNEL staining showed there was no significant apoptosis in the piglet liver ERS model. To explore the biofunction of ER chaperone GRP94, we used shRNA to knock down the expression of GRP94 in porcine HSCs. Interestingly, on porcine HSCs, the knockdown of GRP94 significantly (p < 0.05) decreased the secretion of IGF-1, IGFBP-1 and IGFBP-3 under ERS, but had no significant effect on these under normal condition, and knockdown GRP94 had a significant (p < 0.01) effect on the UBE2E gene and ubiquitin protein from the analysis of two-way ANOVA. On porcine HSCs apoptosis, the knockdown of GRP94 increased the cell apoptosis in TUNEL staining, and the two-way ANOVA analysis shows that knockdown GRP94 had a significant (p < 0.01) effect on the protein levels of Bcl-2 and Caspase-3. For CCK-8 assay, ERS had a significant inhibitory(p < 0.05) effect on cell proliferation when treated with ERS for 24 h, and both knockdown GRP94 and ERS had a significant inhibitory(p < 0.05) effect on cell proliferation when treated with ERS for 36 h and 48 h. We concluded that GRP94 can protect the cell from ERS-induced apoptosis by promoting the IGF-1 system and ubiquitin. These results provide valuable information on the adaptive mechanisms of the liver under ERS, and could help identify vital functional genes to be applied as possible diagnostic biomarkers and treatments for diseases induced by ERS in the future.

Objectives Transport can be a stressful experience for domestic cats. The objective of this study was to evaluate the effect of a new synthetic feline facial pheromone product on relieving stress in domestic cats during short-distance transport. Methods A randomised, blinded, placebo-controlled pilot study was conducted. Prior to baseline screening, cat owners were trained via online meetings in the assessment of stress-related behaviours and overall stress visual analogue scoring. All assessments were completed 30 mins after the start of transport. A total of 150 domestic cats exhibiting stress-related behaviours during baseline screening were recruited and randomly assigned to either the treatment group (n = 75) or the placebo group (n = 75). For the placebo-controlled intervention, the product and placebo, which were identical in appearance, were provided to the treatment and placebo groups, respectively. Fifteen minutes after spraying the carriers with the product or placebo, cat owners were required to take their cat out and complete the same assessments (baseline screening). Results After the intervention, the performance of the stress-related behaviours curling, lack of motion and meowing during transport outside the home was significantly reduced in the treatment group compared with the placebo group ( P 2.94 cm, cats in the treatment group had significantly lower post-intervention stress VAS scores compared with those in the placebo group ( P Conclusions and relevance A synthetic feline facial pheromone product can alleviate short-distance transport-related stress in domestic cats with higher stress scores. This finding will benefit domestic cats during transport outside of the home.

Non-nutritive oral behaviors, especially tongue rolling, are prevalent in the stabled cow population. These behaviors mean that the environment or management process might not suit the cows, suggesting low welfare. However, few researches have reported the physiological indicators or production performance of dairy cows with the stereotyped behavior. This study aimed to determine physical conditions, daily activity, rumen fermentation, and milk production of cows with tongue-rolling behavior. Three hundred and fifty nine Holstein cows in the same barn and lactation stage were subjected to scan sampling behavior observations 126 times for 7 days. Ten cows with high-frequency tongue-rolling behavior (TON) and 10 cows without abnormal oral behavior (CON) were selected for further study. Serum sample, ruminal fluid, milk sample, and behavior record video of TON and CON cows were collected. TON cows had more drinking behavior and more stable lying behavior than the CON cows during the daytime. The body condition score of the TON cows decreased, while the milk yield, yield of milk fat, protein, and lactose in the study period increased. The TON cows had lower ruminal fluid pH, acetate/propionate ratio, and total volatile acid. The bacterial diversity in the ruminal fluid was not different between the two groups. Compared to CON cows, the TON cows had a higher level of serum stress indicators, such as cortisol, thyroid hormone, and norepinephrine, which positively correlated to the frequency of tongue-rolling behavior. Meanwhile, the TON cows had a higher level of lactate dehydrogenase, serum glucose, total triglyceride, total cholesterol, and Interleukin 6. Overall, it means they suffer from higher levels of stress and have higher energy metabolism for a long time when cows show tongue-rolling behavior. TON cows had suffered a higher stress level and had higher energy metabolic status for a long time. The TON cows might have better heat tolerance to the thermal environment by more lying and drinking time. Our data revealed the changes in milk production, physiological stress indicators of dairy cows with high-frequency tongue rolling behavior, which will provide essential knowledge for the in-depth understanding of tongue rolling behavior in dairy cows.

Ammonia (NH3) is a harmful gas with irritating odor, and higher NH3 concentration was found in intensive poultry houses. Although the toxicity of NH3 is well known, little attention has been given to the mechanism of NH3 poisoning in chicken immune organs. To investigate NH3-caused inflammatory damage of broiler spleens, in this study, a broiler model for NH3 poisoning was established, and 3 levels (including microRNA [miRNA], mRNA, and protein) were performed using qRT-PCR and western blot. The results indicated that NH3 exposure caused inflammatory damage using microstructure observation; decreased 2 inflammation-related miRNAs (miR-133a and miR-6615), 2 cytokines secreted by T helper cells 1 (Th1), and heme oxygenase-1 (HO-1); and increased 2 target genes (LOC101747543 and mothers against decapentaplegic homolog 7 [SMAD7]) of the 2 miRNAs, 7 inflammation-related factors, 3 cytokines secreted by Th2, and 5 heat shock proteins (HSPs) in broiler spleens. Our study suggested that Th1/Th2 imbalance, nuclear factor-κB (NF-κB) pathway, and compensatory response of HSPs were involved in NH3-caused inflammatory damage in broiler spleens; there was immunotoxic effect in excess NH3 on broilers. For the first time, we discovered miR-6615 and LOC101747543 may be involved in the mechanism of broiler spleen inflammatory damage caused by NH3 via the NF-κB pathway, and further mechanism needs to be investigated. This study provides new insights for NH3 toxicity identification and risk assessment in animal husbandry production practice.

Background: Lead (Pb) is a toxic environmental pollutant and can exerts toxicity in kidneys. It is known that selenium (Se) has an antagonistic effect on Pb poisoning. However, biological events during the process were not well understood in chicken kidneys.Methods: One hundred and eighty male Hyline chickens (7-day-old) were randomly divided into the control group (offering standard diet and potable water), the Se group (offering Na2SeO3-added standard diet and potable water), the Pb group (offering standard diet and (CH3OO)2Pb-added potable water), and the Pb+Se group (offering Na2SeO3-added standard diet and (CH3OO)2Pb-added potable water). On 30th, 60th, and 90th days, kidneys were removed to perform the studies of histological structure, oxidative stress indicators, cytokines, heat shock proteins, and autophagy in the chicken kidneys.Results: The experimental results indicated that Pb poisoning changed renal histological structure; decreased catalase, glutathione-s-transferase, and total antioxidative capacity activities; increased hydrogen peroxide content; induced mRNA and protein expression of heat shock proteins; inhibited interleukin (IL)-2 mRNA expression, and induced IL-4 and IL-12β mRNA expression; inhibited mammalian target of rapamycin mRNA and protein expression, and induced autophagy-related gene mRNA and protein expression in the chicken kidneys. Supplement of Se mitigated the above changes caused by Pb.Conclusion: Our research strengthens the evidence that Pb induced oxidative stress, inflammation, heat shock response, and autophagy and Se administration alleviated Pb poisoning through mitigating oxidative stress in the chicken kidneys.

This study was undertaken to identify metabolite differences in plasma of dairy cows with a normal or high calving body condition score (CBCS), using untargeted liquid chromatography-mass spectrometry (LC-MS) metabolomics. Sixteen multiparous dairy cows were assigned to one of two groups based on CBCS (0 to 5 scale): Normal group (NBCS, 3.25 &le, BCS &le, 3.5, n = 8), and high BCS group (HBCS, BCS &ge, 4, n = 8). Plasma samples were collected for metabolomics analysis and evaluation of biomarkers of lipid metabolism (nonesterified fatty acid (NEFA) and &beta, hydroxybutyrate (BHB)), and cytokines (leptin, adiponectin, tumor necrosis factor&ndash, &alpha, (TNF-&alpha, ) and interleukin 6 (IL-6)). A total of 23 differential metabolites were identified, and functional analyses were performed using the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Among these metabolites, the concentrations of six lysophosphatidylcholines and one phosphatidylethanolamine, were lower in the HBCS group than in the NBCS group (p &lt, 0.01). Furthermore, these metabolites were involved in these four pathways, among others: glycerophospholipid metabolism, retrograde endocannabinoid signaling, autophagy, and glycosylphosphatidylinositol (GPI)-anchor biosynthesis (p &lt, 0.05). In addition, plasma concentrations of leptin (p = 0.06) and TNF-&alpha, (p = 0.08) tended to be greater while adiponectin (p = 0.09) lower in HBCS cows than in NBCS cows. The concentrations of NEFA, BHB, or IL-6 did not differ between NBCS and HBCS groups. More importantly, based on the results of the Spearman&rsquo, s correlation analysis, the seven important metabolites were negatively correlated with indices of lipid metabolisms, proinflammatory cytokines, and leptin, but positively correlated with adiponectin. These results demonstrate that CBCS has a measurable impact on the plasma metabolic profile, even when NEFA and BHB are not different. In addition, the identified differential metabolites were significantly correlated to lipid metabolism and inflammation in the over-conditioned fresh cows, which are expected to render a metabolic basis for the diseases associated with over-conditioned dry cows.

Aim. To evaluate the role of heat shock protein 70 (HSP70) on the MAPK pathway activation with quercetin treatment and its protection against small intestine impairments of heat stressed rats. Methods. Forty-eight male Sprague-Dawley rats aged 6 weeks were randomized to three groups (n=16/group), namely, control (CON), heat stress (HS), and heat stress + quercetin (HQ). The experiment lasted for 14 days with daily 50 min of heat stress treatment (43°C) for the HS and HQ groups. Rats of HQ group were intragastrically given 0.5 ml quercetin solution (50 mg/kg body weight) before the heat stress treatment. Half of the animals were sacrificed on day 7 and the rest on day 14 for tissue sampling. Intestinal morphology, small intestine morphology and permeability, protein expression of HSP70, phosphorylation of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (MAPK), and caspase-3 activity were examined. Results. Heat stress caused morphological damage to the small intestine and increased intestinal permeability. HSP70 expression and MAPK activity in the small intestine were increased by heat stress. Inhibition of HSP70 by quercetin did not change intestinal permeability compared with the HS group but aggravated intestinal injury and affected the activation of MAPKs and caspase-3. Conclusions. HSP70 may modulate stress-activated signaling and acts in a protective manner via MAPK signaling. Affecting HSP70 protective mechanisms could be useful for protection against heat stress-induced injury in rat small intestine.

Aldehyde dehydrogenase 2 (ALDH2) has both dehydrogenase and esterase activity; its dehydrogenase activity is closely related to the metabolism of aldehydes produced under oxidative stress (OS). In this review, we recapitulate the enzyme activity of ALDH2 in combination with its protein structure, summarize and show the main mechanisms of ALDH2 participating in metabolism of aldehydes in vivo as comprehensively as possible; we also integrate the key regulatory mechanisms of ALDH2 participating in a variety of physiological and pathological processes related to OS, including tissue and organ fibrosis, apoptosis, aging, and nerve injury-related diseases. On this basis, the regulatory effects and application prospects of activators, inhibitors, and protein post-translational modifications (PTMs, such as phosphorylation, acetylation, S-nitrosylation, nitration, ubiquitination, and glycosylation) on ALDH2 are discussed and prospected. Herein, we aimed to lay a foundation for further research into the mechanism of ALDH2 in oxidative stress-related disease and provide a basis for better use of the ALDH2 function in research and the clinic.

Heat stress (HS) has an adverse effect on meat quality; however, the underlying molecular mechanisms altering meat quality due to muscle responses to stress remain unclear. Sixteen castrated male crossbreeds between Landrace × Yorkshire sows and Duroc boars (79.00 ± 1.50 kg body weight) were exposed to either thermal neutral (22 °C, n = 8) or HS (30 °C, n = 8) conditions for 3 weeks. Subsequently, the longissimus dorsi (LD) muscle of all pigs was assayed for meat quality parameters and proteome analysis.; Results: HS decreased post mortem (24 h) pH and intramuscular fat, changed ultimate L*, a* and b* values and increased drip loss and shear force. Proteome analysis of the LD was conducted by two-dimensional gel electrophoresis and mass spectrometry. A total of 23 differentially expressed proteins were identified, of which three were verified by western blotting analysis. The identified proteins were involved in six types of biological process: carbohydrate metabolism, myofibrillar and cytoskeleton structure, stress response, antioxidant and detoxification, calcium binding and cellular apoptosis. Interestingly, HS induced higher levels of heat shock protein, antioxidants and calcium binding proteins, which are involved in the mechanisms of defense and homeostasis.; Conclusion: The results indicate that HS-induced changes in the expression of myofibrillar proteins, glucose and energy metabolism-related proteins, heat shock protein and antioxidant enzymes might, at least partly, contribute to increase in meat tenderness. These findings will provide the foundation for developing future mitigating solutions and preventative therapies to reduce the detrimental effects of chronic HS on muscle function, metabolism and meat quality. © 2017 Society of Chemical Industry.; © 2017 Society of Chemical Industry.

Lead (Pb) is a toxic heavy metal and can harm organisms by inducing apoptosis. Selenium (Se), an essential trace element for humans and animals, can alleviate heavy metal toxicity. The aim of our study is to investigate alleviative effect of Se on Pb-induced apoptosis via endoplasmic reticulum (ER) stress in chicken kidneys. One hundred and eighty male chickens were randomly divided into four groups at 7 days of age and were fed with commercial diet (containing 0.49 mg/kg Se) and drinking water, Na2SeO3-added commercial diet (containing 1 mg/kg Se) and drinking water, the commercial diet and (CH3OO)2Pb-added drinking water (containing 350 mg/L Pb), and Na2SeO3-added commercial diet (containing 1 mg/kg Se) and (CH3OO)2Pb-added drinking water (containing 350 mg/L Pb), respectively. On the 30th, 60th, and 90th days of the experiment period, 15 chickens in each group were euthanized and the kidneys were collected. Following contents were performed: kidney ultrastructure; nitric oxide (NO) content; inducible nitric oxide synthase (iNOS) activity; relative messenger RNA (mRNA) and protein expression of iNOS, ER-related genes (glucose-regulated protein (GRP)78, GRP94, activating transcription factor (ATF)4, ATF6, and iron-responsive element (IRE)), and apoptosis-related genes (caspase-3 and B cell lymphoma-2 (Bcl-2)); and caspase-12 protein expression. The results indicated that Pb changed kidney ultrastructural structure; decreased Bcl-2 mRNA and protein expression; and increased NO content, iNOS activity, relative mRNA and protein expression of iNOS, ER-related genes, and caspase-3 and caspase-12 protein expression. Se attenuated above changes caused by Pb. Pb had time-dependent manners on NO content, GRP78, GRP94, ATF4, IRE, and caspase-3 mRNA expression. Se attenuated Pb-induced apoptosis via ER stress in the chicken kidneys.

The aim of this study was to investigate ameliorative effect of selenium (Se) on lead (Pb)-induced inflammatory damage in chicken testes. One hundred eighty 7-day-old male chickens were randomly assigned into the control group, the Se group, the Pb group, and the Pb/Se group. Lead acetate was added in drinking water (350 mg/L Pb). Sodium selenite was added in the standard commercial diet (1 mg/kg Se). On the 30th, 60th, and 90th days of the experiment, 15 chickens of each group were euthanized. Inductively coupled plasma mass spectrometry, hematoxylin and eosin staining, real-time quantitative PCR, and Western blot were used. The results indicated that excess Pb increased nitric oxide content; inducible nitric oxide synthase (iNOS) activity; nuclear factor-kappa B (NF-κB), tumor necrosis factor-α, cyclooxygenase-2, prostaglandin E synthases, and iNOS mRNA levels in a time-dependent manner; NF-κB, iNOS, heat shock protein (HSP) 60, HSP70, and HSP90 protein levels; and Pb concentration. Excess Pb decreased Se concentration and induced histological changes. Se-alleviated Pb caused all of the above changes. Se improved Pb-caused inflammatory damage by decreasing the expression of inflammatory factors and heat shock proteins in the chicken testes. Our results provided theoretical basis of an alleviative effect of Se on Pb-induced bird testis damage.

We investigated the effect of different farrowing systems on the behavior of sows and piglets. In total, 22 hybrid sows (Yorkshire &times, Landrace) were randomly assigned into three farrowing systems, farrowing crate (FC), free farrowing pen with sloping walls (FFS), and free farrowing pen with sloping walls and nest materials (FFSN). The FFS and FFSN sows were more active, and exhibited less abnormal behaviors compared with the FC sows before and after parturition. FFS and FFSN piglets were more active compared with FC piglets. The increased activity of the FFS and FFSN sows might encourage the activity of their piglets, resulting in more proximity to sow behavior of their piglets. Providing nest materials improved the investigative behavior of sows and piglets. The total mortality of piglets in the free farrowing systems tended to be higher compared with the farrowing crate system. In conclusion, free farrowing system has beneficial effects on the behavior of sows and piglets but extra care in management needs to be taken to avoid piglet loss. Adding nest material in the farrowing pens is suggested to enrich the behavioral pattern of sows and piglets.

Early cow&ndash, calf separation management induced various welfare problems for dairy calves. We mimicked the maternal licking by manually brushing right after the Holstein female calves were born and during their first week of life, termed artificial grooming (AG). The behavior of these treated calves (AG, n = 17) was compared with the calves without artificial grooming (Con, n = 16) during daily behavioral observation around evening milk feeding and in the open field test (OFT) and novel human test (NHT). The number of calves ingesting starter on day six was recorded. The AG calves were observed to be more active and perform more oral behavior compared with the Con calves around evening milk feeding. In the OFT and NHT, the AG calves were again more active than the Con calves. Moreover, the AG calves tended to be less cautious and had more human interactions than the Con calves in the NHT. There tended to be a higher percentage of AG calves ingesting starter on day 6. In conclusion, artificial grooming during early life could boost the activity and the human affinity of female calves and it might advance their starter diet ingestion.

Previous studies suggest that there exists a lag relationship between daily milk yield and heat stress. The values of heat stress indicators (e.g., temperature-humidity index and ambient temperature) before test day have a simple correlation with daily milk yield on test day. However, the simple correlation might not be the best description because daily milk yield and heat stress indicators have a nature of time series in common, and their correlations are cross correlations that could be affected by autocorrelations. We hope to give a more reliable estimation on the lag relationship of daily milk yield via excluding autocorrelations with transfer function modeling. In this study, we found a lag relationship between daily milk yield and heat stress indicators based on transfer function modeling. Heat stress indicators included ambient temperature and temperature-humidity index. The daily milk yield data from 123 cows were obtained during a consecutive 63-d period (July 10-September 10, 2016). The mean daily milk yield (MY) and the maximum daily ambient temperature (TA_max) satisfied the stationary hypothesis, and the cross correlation between them was calculated. Before excluding autocorrelation, MY at 0 to 4 d after test day had significant cross correlations with TA_max on test day. After excluding the influence of autocorrelations, MY at 1 to 3 d after the test day had significant cross correlations with TA_max on test day. This result suggested that MY would respond to TA_max 1 d after the test day. In addition, the strength of cross correlations between MY and TA_max decreased from 1 to 3 d in sequence, implying a declining lag response of MY that would last for 3 d. The transfer function model for MY and TA_max is written as: MYt = 16.90 + 0.74MYt- 1 - 0.25TA_maxt- 1 + Nt, where Nt is white noise. This model can be used to track and predict the dynamic response of MY to TA_max.

Heat stress (HS) negatively affects meat quality by affecting material and energy metabolism, and exploring the mechanism underlying the muscle response to chronic HS in finishing pigs is important for the global pork industry. This study investigated changes in the metabolic profiles of the longissimus dorsi (LD) muscle of finishing pigs under high temperature using ultra-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-ESI-QTOF-MS) and multivariate data analysis (MDA). Castrated male DLY pigs (Duroc &times, Landrance &times, Yorkshire pigs, n = 24) from 8 litters were divided into three treatment groups: constant optimal ambient temperature at 22 &deg, C and ad libitum feeding (CR, n = 8), constant high ambient temperature at 30 &deg, C and ad libitum feeding (HS, n = 8), and constant optimal ambient temperature 22 &deg, C and pair-feeding to the control pigs (PF, n = 8). The metabolic profile data from LD muscle samples were analyzed by MDA and external search engines. Nine differential metabolites (L-carnosine, acetylcholine, inosinic acid, L-carnitine, L-anserine, L-&alpha, glycerylphosphorylcholine, acetylcarnitine, thiamine triphosphate, and adenosine thiamine diphosphate) were involved in antioxidant function, lipid metabolism, and cell signal transduction, which may decrease post mortem meat quality and play important roles in anti-HS. Four metabolites (L-carnosine, acetylcholine, inosinic acid, and L-carnitine) were verified, and it was indicated that the muscle L-carnitine content was significantly lower in HS than in CR (p &lt, 0.01). The results show that constant HS affects the metabolites in the LD muscle and leads to coordinated changes in the endogenous antioxidant defense and meat quality of finishing pigs. These metabonomics results provide a basis for researching nutritional strategies to reduce the negative effects of heat stress on livestock and present new insights for further research.

Heat stress (HS) is a main environmental challenge affecting the animal welfare and production efficiency in pig industry. In recent years, numerous reports have studied the alterations in gene expressions and protein profiles in heat-stressed pigs. However, the use of metabolome to unravel adaptive mechanisms of finishing pig in response to chronic HS have not yet been elucidated. We aimed to investigate the effects of chronic HS on serum metabolome in finishing pigs, and to identify the biomarkers of heat stress. Pigs (n = 8 per treatment) were exposed to either thermal neutral (TN, 22 &deg, C) or heat stress (HS, 30 &deg, C) conditions for three weeks. Serum metabonomics of TN- and HS-treated pigs were compared using the GC-MS approach. Metabonomics analysis revealed that twenty-four metabolites had significantly different levels in TN compared to HS (variable importance in the projection values &gt, 1 and p &lt, 0.05). These metabolites are involved in carbohydrate, amino acid, fatty acid, amines metabolism, and gut microbiome-derived metabolism. Three serum monoses (glucose, mannose 2, and galactose) and 6-phosphogluconic acid were decreased, indicating insufficient source of fuel for energy supply, resulting in negative energy balance (NEB) in heat-stressed pigs. Increased levels of non-esterified fatty acid (myristic acid, palmitic acid, and linoleic acid) and short-chain fatty acids (3-hydroxybutanoic acid and maleic acid) suggested fat decomposition compensating for energy shortage, which was an adaptive response to NEB. Increased concentrations of fluorine, lyxose 1, and D-galacturonic acid were significantly correlated with the levels of acute phase proteins (HP, LBP, &alpha, 2-HSG, and Lysozyme), suggesting acute phase response in HS-stressed pigs. These metabolites are expected to be novel biomarkers of chronic HS in pigs, yet the use of which awaits further validation.

Cadmium (Cd)-caused water environment pollution has become a matter of concern. Gill is an organ with respiratory and mucosal immune functions, and is also one of the organs directly attacked by pollutants. It was found that excess Cd could cause Cd accumulation and gill injury in carp. However, the mechanism of Cd-caused damage in common carp gills is still unclear. Oxidative stress, immunosuppression, and apoptosis took part in the mechanism of poisoning caused by some harmful substances. The aim of the study was to investigate complex molecular mechanism of apoptotic injury caused by Cd in common carp gills. Hence, in this study, we established a Cd poisoning model to explore whether excess Cd can induce apoptosis through observing histomorphology and apoptotic cells; and determining mineral elements, oxidative stress-related factors, immune-related, and apoptosis-related genes in common carp gills. Fifty-four fish were randomly separated into the control group and the Cd group and were cultured for 45 days. The water of the control group was drinking water and the water of the Cd group was CdCl2-added drinking water (0.26 mg/L Cd). In our results, we found that excess Cd increased Cd level, decreased the levels of essential mineral elements (Cu, Fe, Zn, and Mn), damaged mitochondria, and increased apoptotic cells in common carp gills, meaning that excess Cd caused Cd accumulation and apoptotic injury via mitochondrion in common carp gills. Furthermore, we found that Cd inhibited anti-apoptosis-related gene Bcl-2 and stimulated pro-apoptosis-related genes (JNK, FoxO3a, PUMA, Bax, Apaf-1, Caspase-9, and Caspase-3) on 15th, 30th, and 45th days. Above data meant that Cd exposure caused apoptosis via mitochondrion and JNK-FoxO3a-PUMA pathway in common carp gills. In addition, in our experiment, Cd treatment increased oxidants (H2O2 and MDA) and decreased antioxidants (CAT, GPx, GST, SOD, T-AOC, and GSH), indicating that Cd caused oxidative stress via oxidation/antioxidation imbalance. Meanwhile, compared to the control group, T-help 17 (Th17) cell-related factors (IL-17, TNF-α, and RORγ) were up-regulated, regulatory T (Treg) cell-related factors (IL-10 and Foxp3) were down-regulated, and IL-17/IL-10, TNF-α/IL-10, and RORγ/Foxp3 were increased in Cd-exposed group; meaning that excess Cd induced immunosuppression via the imbalance of Th17/Treg cells. Taken together, our findings indicated that JNK-FoxO3a-PUMA pathway and mitochondrion participated in oxidative stress and immunosuppression-mediated apoptosis caused by Cd in common carp (Cyprinus carpio L.) gills. Our data provided new perspectives on the negative effects of heavy metal pollutants on fish.

Lead (Pb) is a toxic metal and can damage chicken organs. Selenium (Se) is an essential micronutrient and has antagonistic effect on heavy metal toxicity. However, the toxic effect of Pb on inflammatory factors and selenoproteins, and antagonistic effect of Se on Pb toxicity in chicken hearts remain unclear. In the present study, a chicken model to study Pb and Se was established. Seven-day-old male chickens were randomly divided into the control, the +Se group, the +Pb group, and the Se + Pb group. The feeding program of Pb and Se was as follows: lead acetate was added to drinking water and sodium selenite was added to the standard diet. Relative mRNA levels of four inflammatory factors (nuclear factor-κB (NF-κB), tumor necrosis factor-α, cyclooxygenase-2, and prostaglandin E synthases) and 25 selenoproteins (including glutathione peroxidase 1 (Gpx1), Gpx2, Gpx3, and Gpx4, thioredoxin reductase 1 (Txnrd1), Txnrd2, Txnrd3, iodothyronine deiodinase 1 (Dio1), Dio2, Dio3, selenoprotein n1 (Sepn1), selenoprotein K (Selk), Sels, Sepw1, Selt, Selh, Selm, 15 kDa selenoprotein, Seli, Selu, Selpb, Sepp1, Selo, Sepx1, selenophosphate synthetase 2 (SPS2)) were detected on the 30th, 60th, and 90th days in chicken hearts. The results indicated that Pb poisoning increased mRNA levels of inflammatory factors, decreased mRNA levels of selenoproteins, and caused histological changes in the chicken hearts. Se alleviated Pb-caused all the above changes in the chicken hearts. Our results suggested that Se supplementation alleviated Pb-induced inflammatory injury through inhibiting the NF-κB signaling pathway and stimulating selenoproteins in the chicken hearts.

Milk production and time effects are considered related to heat stress but they have not yet been combined in predictive models. In two parts, this study aimed to develop new models to predict heat stress (rectal temperature and respiration rate) of lactating dairy cows by inputting predictors, including ambient temperature (Ta), relative humidity (RH), wind speed (WS), milk yield (MY), and time blocks. In the first part of the study, we built the quantitative foundation for the second part, including the regression relation between respiration rate and rectal temperature (to convert predicted respiration rate to predicted body temperature), as well as between rectal temperature and respiration rate when heat stress was triggered (to recognize whether herds were under stress). In the second part, we built models that combined the abovementioned predictors to predict respiration rate. In part I, data were obtained from 45 high-producing Holstein cows within a Ta range of 9.5 to 30.8°C. We found a very strong correlation between mean respiration rate (MRR) and mean rectal temperature (MRT), where MRT = 0.021 × MRR + 37.6 (R2 = 0.925), suggesting that for each 4.8 breaths per minute (bpm) increase of MRR, MRT would be expected to increase by 0.1°C. Rectal temperature was determined to be 38.6°C when heat stress was triggered, which corresponded to a respiration rate of 48 bpm. In part II, data were obtained in 3 stalls within a Ta range of 6.9 to 33.3°C over 3 time blocks, all of which were the 90 min preceding milking (0630-0800, 1230-1400, and 1830-2000 h). We found a nonlinear response of MRR to Ta, which could be linearized by the quadratic term of Ta. The response of MRR was the highest in the 0630-0800 h block, followed by 1230-1400 h, and finally 1830-2000 h. We proposed a model combining 3 time blocks (R2 = 0.836): MRR in 0630-0800 h was determined to 56.28 + (-3.40 + 0.11 × Ta + 0.02 × RH) × Ta - 0.21 × RH - 2.82 × WS + 0.62 × MY; MRR in 1230-1400 h and 1830-2000 h were 4.6 and 10.3 bpm lower than that in 0630-0800 h, respectively (reducing the intercept of the expression in 0630-0800 h). Compared with temperature-humidity index equations, the proposed model performed better at suppressing prediction error, and had better sensitivity and accuracy in recognizing whether heat stress was triggered.

The microbiome has emerged as a new player on behavior, physiology and stress because of its significant effects on the brain-gut axis. The aim of this study was to increase our understanding of brain-gut function in dairy cows. We investigated the effects of a heat-stress (HS) environment and individual differences of heat sensitivity (IH) on bovine physiological characteristics and microbial composition. Results indicate that both HS and IH increased rectal temperature (RT) (P P P P

The objectives of the research reported in this Research Communication were to compare the variation of hind quarter skin surface temperature pre- and post- milking in dairy cows and to determine the optimal time to capture images by infrared thermography for improving the sensitivity and specificity of mastitis detection in dairy cows. Hind quarter infrared images of 102 Holstein dairy cows were captured from the caudal view by an infrared camera pre-milking and post-milking. The udder skin surface temperature was measured with the help of the image processing software. No significant difference was found between the left and right quarter skin surface temperature pre- and post- milking. The hind quarter skin surface temperature pre-milking was not significantly influenced by milk yield, but exhibited a rising trend along with the increase of milk yield. The hind quarter skin surface temperature post-milking was significantly influenced by milk yield. This leads us to conclude that the sensitivity and specificity of IRT in mastitis detection may be influenced by milk yield and it may be better to capture the infrared images of cow udders pre-milking.

Acute heat stress (HS) negatively affects intestinal integrity and barrier function. In contrast, chronic mild HS poses a distinct challenge to animals. Therefore, this study integrates biochemical, histological and proteomic approaches to investigate the effects of chronic HS on the intestine in finishing pigs. Castrated male crossbreeds (79.00±1.50 kg BW) were subjected to either thermal neutral (TN, 21 °C; 55%±5% humidity; n=8) or HS conditions (30 °C; 55%±5% humidity; n=8) for 3 weeks. The pigs were sacrificed after 3 weeks of high environmental exposure and the plasma hormones, the intestinal morphology, integrity, and protein profiles of the jejunum mucosa were determined. Chronic HS reduced the free triiodothyronine (FT3) and GH levels. HS damaged intestinal morphology, increased plasma d-lactate concentrations and decreased alkaline phosphatase activity of intestinal mucosa. Proteome analysis of the jejunum mucosa was conducted by 2D gel electrophoresis and mass spectrometry. Fifty-three intestinal proteins were found to be differentially abundant, 18 of which were related to cell structure and motility, and their changes in abundance could comprise intestinal integrity and function. The down-regulation of proteins involved in tricarboxylic acid cycle (TCA cycle), electron transport chain (ETC), and oxidative phosphorylation suggested that chronic HS impaired energy metabolism and thus induced oxidative stress. Moreover, the changes of ten proteins in abundance related to stress response and defense indicated pigs mediated long-term heat exposure and counteracted its negative effects of heat exposure. These findings have important implications for understanding the effect of chronic HS on intestines.

The insulin-like activity of insulin-like growth factor-1 (IGF-1) is heavily blunted by IGF binding proteins (IGFBPs, including IGFBP-1 and IGFBP-3). The effects of heat stress (HS) on the IGF-1 and IGFBPs in growing pigs are still not clear. This study aimed to investigate the alterations of IGF-1 and IGFBPs under chronic HS in growing pigs. Twenty-seven growing large white barrows with similar body weight were collected from nine litters and were assigned into three treatments. The litter effect is balanced in all treatments. Treatments were: 1) thermal-neutral (TN) group (23 °C), 2) chronic HS group (33 °C), and 3) pair-fed in TN condition (PFTN). The experiment lasted for 21 days. Compared with TN controls, decreased FI, lower average body weight gain, higher rectal temperature and increased respiration rates were observed in HS pigs. On D7, increased plasma insulin concentration and insulin:glucose ratio were observed in HS pigs compared to TN controls. A overall elevation of plasma IGF-1:IGFBP-3 ratio was detected in HS pigs compared with PFTN and TN counterparts. Besides, hepatic IGF-1 gene expression of HS pigs was 50% higher than TN counterparts. PFTN pigs, however, had no differences in plasma IGF-1:IGFBP-3 ratio and hepatic IGF-1 gene expression, compared with TN pigs. PFTN pigs increased plasma IGFBPs concentration and hepatic IGFBPs gene expression, compared with TN controls. However, no differences in plasma IGFBPs concentration and hepatic IGFBPs gene expression were observed between TN and HS group. Liver IRS-1 gene and protein expressions of HS pigs tended to be increased compared with TN controls, while PFTN pigs still kept liver IRS-1 gene and protein expressions in normal level. High temperature (33 °C), independent of feed intake reduction, increased IGF-1 but did not affect IGFBPs levels in growing pigs.

Heat stress (HS) has an adverse effect on meat quality; however, the underlying molecular mechanisms altering meat quality due to muscle responses to stress remain unclear. Sixteen castrated male crossbreeds between Landrace × Yorkshire sows and Duroc boars (79.00 ± 1.50 kg body weight) were exposed to either thermal neutral (22 °C, n = 8) or HS (30 °C, n = 8) conditions for 3 weeks. Subsequently, the longissimus dorsi (LD) muscle of all pigs was assayed for meat quality parameters and proteome analysis.HS decreased post mortem (24 h) pH and intramuscular fat, changed ultimate L*, a* and b* values and increased drip loss and shear force. Proteome analysis of the LD was conducted by two-dimensional gel electrophoresis and mass spectrometry. A total of 23 differentially expressed proteins were identified, of which three were verified by western blotting analysis. The identified proteins were involved in six types of biological process: carbohydrate metabolism, myofibrillar and cytoskeleton structure, stress response, antioxidant and detoxification, calcium binding and cellular apoptosis. Interestingly, HS induced higher levels of heat shock protein, antioxidants and calcium binding proteins, which are involved in the mechanisms of defense and homeostasis.The results indicate that HS-induced changes in the expression of myofibrillar proteins, glucose and energy metabolism-related proteins, heat shock protein and antioxidant enzymes might, at least partly, contribute to increase in meat tenderness. These findings will provide the foundation for developing future mitigating solutions and preventative therapies to reduce the detrimental effects of chronic HS on muscle function, metabolism and meat quality. © 2017 Society of Chemical Industry.

The objective of this study was to assess the effects of constant high ambient temperatures on meat quality, antioxidant capacity, and carnosine expression in longissimus dorsi muscle of finishing pigs. Castrated 24 male DLY (crossbreeds between Landrace×Yorkshire sows and Duroc boars) pigs were allocated to one of three treatments: constant ambient temperature at 22°C and ad libitum feeding (CON, n = 8); constant high ambient temperature at 30°C and ad libitum feeding (H30, n = 8); and constant ambient temperature at 22°C and pair-fed with H30 (PF, n = 8). Meat quality, malondialdehyde (MDA) content, antioxidant capacity, carnosine content, and carnosine synthetase (CARNS1) mRNA expression in longissimus dorsi muscle were measured after three weeks. The results revealed that H30 had lower pH24 h, redness at 45 min, and yellowness at 24 h post-mortem (p

Heat stress affects muscle development and meat quality in food animals; however, little is known regarding its regulatory mechanisms at the epigenetic level, such as via DNA methylation. In this study, we aimed to compare the DNA methylation profiles between control and heat-stressed pigs to identify candidate genes for skeletal muscle development and meat quality. Whole-genome bisulfite sequencing was used to investigate the genome-wide DNA methylation patterns in the longissimus dorsi muscles of the pigs. Both groups showed similar proportions of methylation at CpG sites but exhibited different proportions at non-CpG sites. A total of 57,147 differentially methylated regions were identified between the two groups, which corresponded to 1,422 differentially methylated genes. Gene ontogeny and KEGG pathway analyses indicated that these were mainly involved in energy and lipid metabolism, cellular defense and stress responses and calcium signaling pathways. This study revealed the global DNA methylation pattern of pig muscle between normal and heat stress conditions. The result of this study might contribute to a better understanding of epigenetic regulation in pig muscle development and meat quality.

Heat stress (HS) negatively affects human health, animal welfare, and livestock production. We analyzed the hepatic proteomes of finishing pigs subjected to chronic heat stress (HS), thermal neutral (TN), and restricted feed intake conditions, identifying differences between direct and indirect (via reduced feed intake) HS. Twenty-four castrated male pigs were randomly allocated to three treatments for three weeks: (1) thermal neutral (TN) (22 °C) with ad libitum feeding; (2) chronic HS (30 °C) with ad libitum feeding; and (3) TN, pair-fed to HS intake (PF). Hepatic proteome analysis was conducted using two-dimensional gel electrophoresis and mass spectrometry. Both HS and PF significantly reduced liver weight (p < 0.05). Forty-five hepatic proteins were differentially abundant when comparing HS with TN (37), PF with TN (29), and HS with PF (16). These proteins are involved in heat shock response and immune defense, oxidative stress response, cellular apoptosis, metabolism, signal transduction, and cytoskeleton. We also observed increased abundance of proteins and enzymes associated with heat shock response and immune defense, reduced the redox state, enhanced multiple antioxidant abilities, and increased apoptosis in HS liver. Heat-load, independent of reduced feed intake, induced an innate immune response, while food restriction caused stress and cellular apoptosis. Our results provide novel insights into the effects of chronic HS on liver.

The aim of the current study was to investigate the effect of a permanent heat exposure during 21 days on pig performance, nutrient digestibility, physiological response and key enzyme of skeletal muscle energy metabolism. Twenty-four male finishing pigs (crossbreed castrates, 79.0 ± 1.50 kg body weight) were allocated to three groups (n = 8): (1) Control (ambient temperature (AT) 22°C, ad libitum feeding), (2) Group HE (AT 30°C, ad libitum feeding) and (3) Group PF (AT 22°C, pair-fed to Group HE). The permanent heat exposure decreased feed intake (p < 0.01), daily body weight gain (p < 0.05) and the digestibility of gross energy, dry matter, crude protein and ash (p < 0.05); rectal temperature and respiration rate were significantly increased (p < 0.01). The levels of plasma cortisol, creatine kinase and lactate dehydrogenase were also significantly increased in Group HE (p < 0.05). Furthermore, the heat exposure changed intracellular energy metabolism, where the AMP-activated protein kinase was activated (p = 0.02). This was combined with changes in parameters of glycolysis such as an accumulation of lactic acid (p = 0.02) and a drop of pH24 h (p = 0.02), an increase of hexokinase and pyruvate kinase activity (p < 0.01) and, finally, the maturation process of post mortem muscle was influenced. Due to pair-feeding it was possible to evaluate the effects of heat exposure, which were not dependent on reduced feed intake. Such effects were, e.g., reduced nutrient digestibility and changed activities of several enzymes in muscle and blood serum.