Ginseng Extract Ameliorates the Negative Physiological Effects of Heat Stress by Supporting Heat Shock Response and Improving Intestinal Barrier Integrity: Evidence from Studies with Heat-Stressed Caco-2 Cells, C. elegans and Growing Broilers.

Climatic changes and heat stress have become a great challenge in the livestock industry, negatively affecting, in particular, poultry feed intake and intestinal barrier malfunction. Recently, phytogenic feed additives were applied to reduce heat stress effects on animal farming. Here, we investigated the effects of ginseng extract using various in vitro and in vivo experiments. Quantitative real-time PCR, transepithelial electrical resistance measurements and survival assays under heat stress conditions were carried out in various model systems, including Caco-2 cells, Caenorhabditis elegans and jejunum samples of broilers. Under heat stress conditions, ginseng treatment lowered the expression of HSPA1A (Caco-2) and the heat shock protein genes hsp-1 and hsp-16.2 (both in C. elegans), while all three of the tested genes encoding tight junction proteins, CLDN3, OCLN and CLDN1 (Caco-2), were upregulated. In addition, we observed prolonged survival under heat stress in Caenorhabditis elegans, and a better performance of growing ginseng-fed broilers by the increased gene expression of selected heat shock and tight junction proteins. The presence of ginseng extract resulted in a reduced decrease in transepithelial resistance under heat shock conditions. Finally, LC-MS analysis was performed to quantitate the most prominent ginsenosides in the extract used for this study, being Re, Rg1, Rc, Rb2 and Rd. In conclusion, ginseng extract was found to be a suitable feed additive in animal nutrition to reduce the negative physiological effects caused by heat stress.

In vitro supplementation of trans-10, cis-12 conjugated linoleic acid ameliorated deleterious effect of heat stress on bovine oocyte developmental competence.

Environmental stresses, such as heat stress (HS), have been shown to have diverse effects on the developmental competence of oocytes. The aim of this study was to determine the effect of exogenous conjugated linoleic acid (CLA) supplementation in maturation medium on bovine oocyte maturation and developmental competence under HS condition. Accordingly, cumulus-oocyte complexes (COCs) were cultured at 41 °C and 38.5 °C for the first and second 12 h of maturation in the presence of 0 (PC), 50 (CLA50-HS) and 100 (CLA100-HS) µM CLA. Also, a group of COCs were cultured at 38.5 °C for 24 h of maturation without CLA supplementation as negative control (NC). Nuclear maturation, level of intracellular glutathione (GSH), reactive oxygen species (ROS) content, cleavage and blastocyst rates as well as relative expression of BAX, and BCL2 genes in blastocysts were investigated. Our finding for the PC and NC groups revealed that HS decreased the percentage of MII oocytes, cleavage and blastocyst rates (P < 0.05). Moreover, HS lead to an increase in ROS levels and relative expression of BAX gene, decreased the intracellular content of GSH and relative expression of BCL2 gene (P < 0.05). However, the cleavage and blastocyst rates tended to increase in the CLA-supplemented groups compared to PC group (p < 0.10). Also, ROS and GSH levels in the matured oocytes decreased and increased in the CLA50-HS group compared to the PC group (P < 0.05), respectively. The ratio of expression levels of BAX to BCL2 genes was not different between the PC and CLA50-HS groups (P > 0.05). These findings suggest that HS has undesirable effects on the maturation competence of bovine oocyte and subsequent embryo development while administration of CLA can ameliorate some of adverse effects of HS.

Effect of curcumin supplementation on TLR4 mediated non-specific immune responses in liver of laying hens under high-temperature conditions.

The liver performs a significant role in innate and adaptive immunity. Heat stress causes oxidative stress in liver tissues and reduces the immune responses of laying hens which can cause several diseases affecting poultry-production performance. Hepatic inflammation is a common trigger of liver disease, which is reflected by hepatic tissue damage leading to fibrogenesis and hepatocellular carcinoma. Dietary manipulation of curcumin has been proposed to ameliorate the immune status of chickens under heat stress. Thus, this study aimed to investigate the effect of curcumin supplementation on TLR4 mediated non-specific immune response in liver of laying hens under high-temperature conditions. Experimental groups contained two controls groups (high temperature and thermo-neutral control (HC and NC) fed basal diet) and three high-temperature curcumin treatments groups (HT100, HT200 and HT300). Laying hens in HC and HT groups exposed 6 h/day heat stress (32 ±â€¯1 °C). The results of present study showed that heat stress curcumin treatment group had reduced inflammatory responses (IL-6, IL-1ß, TNF-α) as compared to HC and NC group. Pathological lesions and DNA damage of immune tissues were decreased in heat stress curcumin supplementation as compared to HC and NC group. Furthermore, PCNA, TLR4 and its downstream gene expression as well as protein expression (TLR4, NF-κB and PCNA) were significantly down regulated in heat stress curcumin supplemented group as compared to HC and NC group. Therefore, it is concluded that heat stressed hens supplemented with dietary curcumin enhance the immunity of laying hens and combat stressful environmental conditions.

Experimental heat stress nephropathy and liver injury are improved by allopurinol.

An epidemic of chronic kidney disease (CKD) has been observed in Central America among workers in the sugarcane fields. One hypothesis is that the CKD may be caused by recurrent heat stress and dehydration, and potentially by hyperuricemia. Accordingly, we developed a murine model of kidney injury associated with recurrent heat stress. In the current experiment, we tested whether treatment with allopurinol (a xanthine oxidase inhibitor that reduces serum urate) provides renal protection against recurrent heat stress and dehydration. Eight-week-old male C57BL/6 mice were subjected to recurrent heat stress (39.5°C for 30 min, 7 times daily, for 5 wk) with or without allopurinol treatment and were compared with control animals with or without allopurinol treatment. Mice were allowed ad libitum access to normal laboratory chow (Harlan Teklad). Kidney histology, liver histology, and renal function were examined. Heat stress conferred both kidney and liver injury. Kidneys showed loss of proximal tubules, infiltration of monocyte/macrophages, and interstitial collagen deposition, while livers of heat-stressed mice displayed an increase in macrophages, collagen deposition, and myofibroblasts. Allopurinol provided significant protection and improved renal function in the heat-stressed mice. The renal protection was associated with reduction in intrarenal uric acid concentration and heat shock protein 70 expression. Heat stress-induced renal and liver injury can be protected with allopurinol treatment. We recommend a clinical trial of allopurinol for individuals developing renal injury in rural areas of Central America where the epidemic of chronic kidney disease is occurring.

Nanoselenium Supplementation of Heat-Stressed Broilers: Effects on Performance, Carcass Characteristics, Blood Metabolites, Immune Response, Antioxidant Status, and Jejunal Morphology.

An experiment was conducted to investigate the effects of dietary nanoselenium supplementation at 0, 0.6 and 1.2 mg/kg of diet on growth performance, serum biochemical parameters, immune response, antioxidant capacity, and jejunal morphology of 29-d-old male broilers subjected to heat stress at 37 ± 1°C for 14 d. Broilers were fed for 42 d on the experimental diets. The results showed that nanoselenium supplementation had no effect on growth performance, but it supplementation at the rate of 1.2 mg/kg diet decreased the serum concentration of cholesterol prior to the heat exposure. Further, dietary nanoselenium supplementation linearly increased the high-density lipoprotein cholesterol concentration, while linearly decreased those of low-density lipoprotein cholesterol and aspartate aminotransferase in the serum before applying heat stress. Compared with thermoneutral temperature, heat stress reduced body mass gain, feed intake, percentages of carcass, breast, leg, abdominal fat, bursa of Fabricius, thymus, antibody response against sheep red blood cells, serum concentration of protein, erythrocyte activities of glutathione peroxidase and superoxide dismutase, jejunal villus height, and villus height to crypt depth ratio, while increased feed conversion ratio, percentages of liver, gizzard, pancreas, gallbladder, heart, and the concentrations of aspartate aminotransferase and malondialdehyde. Dietary supplementation of nanoselenium linearly reduced the abdominal fat and liver percentages, while linearly increased the activity of glutathione peroxidase and villus height in heat-stressed broilers. Furthermore, the lower level of nanoselenium decreased the percentages of gizzard and heart in broilers under heat stress. The diet supplemented with 1.2 mg/kg nanoselenium improved feed conversion ratio and increased antibody response against sheep red blood cells, activity of superoxide dismutase, and villus height to crypt depth ratio, but decreased the serum concentrations of cholesterol, low-density lipoprotein cholesterol, and malondialdehyde in heat-stressed broilers. The results suggest that supplemental nanoselenium improved growth performance, internal organs health, immune response, and jejunal morphology by alleviating the oxidative stress induced by heat stress.

Mitigation of heat stress-related complications by a yeast fermentate product.

Heat stress results in a multitude of biological and physiological responses which can become lethal if not properly managed. It has been shown that heat stress causes significant adverse effects in both human and animals. Different approaches have been proposed to mitigate the adverse effects caused by heat stress, among which are special diet and probiotics. We characterized the effect of the yeast fermentate EpiCor (EH) on the prevention of heat stress-related complications in rats. We found that increasing the body temperature of animals from 37.1±0.2 to 40.6±0.2°C by exposure to heat (45°C for 25min) resulted in significant morphological changes in the intestine. Villi height and total mucosal thickness decreased in heat-stressed rats pre-treated with PBS in comparison with control animals not exposed to the heat. Oral treatment of rats with EH before heat stress prevented the traumatic effects of heat on the intestine. Changes in intestinal morphology of heat-stressed rats, pre-treated with PBS resulted in significant elevation of lipopolysaccharides (LPS) level in the serum of these animals. Pre-treatment with EH was effective in the prevention of LPS release into the bloodstream of heat-stressed rats. Our study revealed that elevation of body temperature also resulted in a significant increase of the concentration of vesicles released by erythrocytes in rats, pre-treated with PBS. This is an indication of a pathological impact of heat on the erythrocyte structure. Treatment of rats with EH completely protected their erythrocytes from this heat-induced pathology. Finally, exposure to heat stress conditions resulted in a significant increase of white blood cells in rats. In the group of animals pre-treated with EH before heat stress, the white blood cell count remained the same as in non-heated controls. These results showed the protective effect of the EH product in the prevention of complications, caused by heat stress.

Heat shock stress prior to hyperbaric oxygen exposure in rats.

INTRODUCTION: Heat shock proteins (HSPs) and nuclear factor-kappa B (NF-kappaB) have been established as important mediators in lung injury; however, their role in preventing pulmonary toxicity from hyperbaric oxygen (HBO) has not been evaluated. METHODS: We aimed to study the effects of heat shock (HS) injury on hyperbaric hyperoxic lung injury (HHLI) in a rat model and identify a mechanism of protection by evaluating HSP 27 and HSP 70 mRNA and protein levels, NF-kappaB p65, lung injury and oxidative parameters. By varying the times between HS and exposure to HBO, the pathways of interaction between HSPs and NF-kappaB will be further clarified. RESULTS: Our results showed that HS exposure increases the mRNA and protein levels of HSP 27 and HSP 70; HS induced 10-fold increases of HSP 27 (9.77 +/- 0.60) and HSP 70 (10.33 +/- 2.4) within the first 10 h compared to control animals. Lesion scores were higher for the first 16 h after HS, but decreased again after 31 h (N = 7 animals; 5 lesions scores). Protein nitration showed no significant differences between groups exposed to HS or HBO; similarly there was no difference with a combination of both treatments. DISCUSSION: HBO appears to attenuate the HS response by HSP 27 and HSP 70. Histopathology results suggest that HS might mitigate pathology in animals exposed to HS and HBO. No significant effect of HS on HBO-induced HHLI was observed in animals treated with both HS and HBO.

Performance, egg quality, and immune response of laying hens fed diets supplemented with mannan-oligosaccharide or an essential oil mixture under moderate and hot environmental conditions.

In total, 432 thirty-six-week-old laying hens were fed a basal diet supplemented with mannan-oligosaccharide (MOS) or an essential oil mixture (EOM) from 36 to 51 wk of age. Hens were divided into 3 equal groups replicated 6 times with 24 hens per replicate. No significant difference was observed among the dietary treatments in terms of performance indices. Different from the dietary manipulation, high environmental temperatures negatively influenced all of the laying performance traits except the feed conversion ratio in association with the diminished feed consumption. The MOS, and particularly the EOM, tended to alleviate the deleterious effect of heat stress on BW gain. Mortality was higher in MOS-fed hens than with other treatments. A supplementation diet with MOS or EOM provided increments in eggshell weight (P < 0.01). Relative albumen weight was significantly decreased (P < 0.05) in response to EOM or MOS supplementation; however, this was not the case in the yolk weight rate. The MOS decreased albumen height and Haugh unit (P < 0.05). High environmental temperatures hampered entire egg quality characteristics except for the eggshell breaking strength and egg yolk weight. These results indicated that heat stress adversely affected both productive performance and egg quality. As for the results of this study, neither MOS nor EOM was efficacious in improving efficiency of egg production and stimulating humoral immune response in laying hens reared under moderate and hot climatic conditions. However, the ameliorative effect exerted by MOS and EOM on eggshell characteristics is conclusive.

Dysregulation of hepatic iron with aging: implications for heat stress-induced oxidative liver injury.

Environmental heat stress is associated with an age-related increase in hepatic oxidative damage and an exaggerated state of oxidative stress. The purpose of this investigation was to evaluate the regulation of hepatic iron after heat stress. A secondary aim was to determine a potential role for iron in heat stress-induced liver injury. Hyperthermia-induced alterations in hepatic iron were evaluated in young (6 mo) and old (24 mo) Fischer 344 rats by exposing them to a two-heat stress protocol. Livers were harvested at several time points after the second heating and assayed for labile and nonheme iron. In the control condition, there was no difference in labile iron between age groups. Both labile iron and storage iron were not altered by hyperthermia in young rats, but both were increased immediately after heating in old rats. To evaluate a role for iron in liver injury, hepatic iron content was manipulated in young and old rats, and then both groups were exposed to heat stress. Iron administration to young rats significantly increased hepatic iron content and ferritin but did not affect markers of lipid peroxidation under control conditions or after heat stress. In old rats, iron chelation with deferoxamine prevented the increase in nonheme iron, labile iron, ferritin, and lipid peroxidation after heat stress. These results suggest that iron may play a role in hepatic injury after hyperthermia. Thus, dysregulation of iron may contribute to the gradual decline in cellular and physiological function that occurs with aging.

[Observation of sanxian xinli capsule on pathologic morphology in superheat mice].

OBJECTIVE: To investigate the change of pathologic morphology in superheat mice. METHOD: The superheat was used in the experinent. The pathologic morphology of thymus gland, spleen, liver and lung was observed by light microscope. RESULT: The histiocytes of thymus gland, spleen, liver and lung presented no changes in lesion, congestion and morphology. CONCLUSION: The Sanxian Xinli capsule have function of protecting the superheat mouse viscera.

Heat stress induced histopathology and pathophysiology of the central nervous system.

The number of reports on the effects of heat stress is still increasing on account of the temperature is one of the most encountered stressful factors on the different biological systems. Because the heat stress (HS) considered a model of thermal injury to the central nervous system (CNS), the purpose of this review was to assess the histopathological changes of HS on CNS. Also, this review emphasized that the heat stress may retard partially the degree of the postnatal neurogenesis and growth of CNS. Taken together, owing to one of the most important functions of heat shock protein is to protect the organisms from the deleterious effects of temperature, thus, it can be hypothesized that the formation of heat shock proteins may be related to the deleterious effect of HS. On the other hands, the alterations of neurotransmitters in the central nervous system might be involved in the physiological and biochemical responses that occur during heat stress. The hypothalamic monoaminergic systems play an important role in the thermoregulation through regulate the heat production and heat dissipation. In addition, the disturbance in the biochemical variables due to the high temperature may be the cause of the histopathological changes and the partial retardation in CNS and the reverse is true. Thus, further studies need to be done to emphasize this concept.

Hyperthermia-induced proteasome inhibition and loss of androgen receptor expression in human prostate cancer cells.

Prostate cancer is the second leading cause of death in men in western countries and is usually treated by surgery and/or radiotherapy. More recently, hyperthermia has been introduced into clinical trials investigating a possible effect in the first-line treatment of prostate cancer. However, the molecular mechanisms of hyperthermia are not completely understood. In this study, we investigated the effects of hyperthermia on proteasome function and its significance for signal transduction, cell death and androgen receptor (AR) expression in PC-3, LnCaP, and DU-145 human and TRAMP-C2 murine prostate cancer cells. Hyperthermia caused apoptosis and radiosensitization and decreased 26S proteasome activity in all three human cell lines to about 40% of untreated control cells. 20S proteasome activity was not affected by heat. Heat treatment inhibited constitutive and radiation-induced activation of nuclear factor kappaB caused by stabilization of IkappaB. Although stabilization of AR by proteasome inhibitors has been reported previously, AR protein levels in LnCaP cells decreased dramatically after heat. Our data suggest that inhibition of proteasome function and dependent signal transduction pathways might be a major molecular mechanisms of heat-induced apoptosis and radiosensitization. Hyperthermia abrogates AR expression in androgen-dependent cells and might thus promote malignant progression of prostate cancer.

Quercetin, apoptosis, heat shock.

The present study was designed to investigate the correlation between the expression level of Hsp27 and Hsp72 and induction of apoptosis in HeLa cells in response to quercetin treatment. Treatment of HeLa cells with quercetin or with 1hr period of hyperthermia (42 degrees) increased the number of apoptotic cells. Inhibition of the expression of Hsp72 and Hsp27 in tumour cells by anti-sense oligonucleotides, enhanced the induction of apoptosis by quercetin. Heat shock itself had little effect on apoptotic cell death in these cells, but when combined with quercetin treatment, caused a significant increase in the number of apoptotic cells. These results suggest that the reduction of Hsps expression in the HeLa cell line promotes the induction of apoptosis by quercetin.