Innate immune and chronic heat stress responses in sturgeons: Advances and insights from studies on Russian sturgeons.

Chronic stress deteriorates the immune function of fish, thereby increasing their vulnerability to infections. However, the molecular and cellular mechanisms underlying stress-mediated immunosuppression and infection susceptibility in fish remain largely unknown. Understanding these mechanisms will contribute to improving fish welfare and their farm production. Herein, we review the challenges of sturgeon aquaculture in subtropical countries, where current climate change has giving rise to significant temperature increments during summer. This leads to the exposure of fish to stressful conditions during these months. Chronic heat stress deserves attention considering the rapid warming rate of the planet. It is already affecting wild fish populations, with disastrous consequences for sturgeons, which are one of the most endangered fish species in the world. In this context, we discuss the most recent advances through the studies on the effects of chronic heat stress on the innate immune components of sturgeons. To this end, we summarise the findings of studies focusing on the aquaculture of Russian sturgeons and observations made on other Acipenser species. Special attention is given to acute-phase proteins, as they might be valuable biomarkers of heat stress and infection, with applicability in monitoring the fish health status in farms.

Effects of lowering dietary protein content without or with increased protein-bound and feed-grade amino acids supply on growth performance, body composition, metabolism, and acute-phase protein of finishing pigs under daily cyclic heat stress.

This study investigated the effects of a low-protein diet with or without an increase in dietary protein and feed-grade amino acids (AAs) on the growth performance, body composition, metabolism, and serum acute-phase proteins of finishing pigs reared in thermoneutrality or cyclic heat stress conditions. A total of 90 gilts (67.7 ± 6.2 kg) were distributed in a 2 × 3 factorial arrangement (two ambient temperatures and three diets). Ambient temperatures (AT) were thermoneutral (TN, 22 °C for 24 h) and cyclic heat stress (CHS, 12 h to 35 °C and 12 h to 22 °C). The evaluated diets (D) were high crude protein (HP); low CP-free AA-supplemented diets (LPAAs); low CP-free AA-supplemented diets and digestible Lys level (+20%), and Lys:AA ratios above recommendations (LPAA+). The experimental period lasted 48 d (two experimental phases: days 0-27 and days 28-48, respectively). CHS pigs had higher skin temperature (P < 0.05) than TN pigs. Pigs in CHS had higher rectal temperature (P < 0.05) than TN pigs until day 38 but similar (P > 0.10) to TN pigs from 38 to 45 d. For the entire experiment, CHS pigs had lower (P < 0.05) final BW, average daily gain and daily feed intake, net energy intake, body lipid, bone mineral, lipid deposition, energy retention, Lys and CP intake, and nitrogen excretion than TN pigs. The level of CP intake impacted nitrogen excretion, nitrogen retention efficiency, and urea as pigs fed HP had the highest values, and pigs fed LPAA had the lowest values (P < 0.05). On day 27, CHS pigs had lower (P < 0.05) free triiodothyronine than TN pigs. LPAA+ pigs had lower (P < 0.05) insulin than LPAA. On day 48, CHS pigs had lower (P < 0.05) thyroxine, albumin, and lactate than TN pigs. On day 27, pigs fed LPAA+ had higher (P < 0.05) lactate than pigs fed HP or LPAA. Both AT and D were enough to stimulate the immune system as CHS pigs had lower (P < 0.05) transferrin and 23-kDa protein levels than TN pigs, and HP pigs had higher haptoglobin than LPAA on day 27. These results confirm the deleterious effects of high AT on performance, body composition, metabolism, and immune system stimulation in finishing pigs. These data also show that a diet with low levels of CP can be provided to pigs in CHS without affecting performance and body composition while reducing nitrogen excretion. However, the use of a diet with an AA level above the requirements obtained by increasing intact protein and free AA did not attenuate the impact of CHS on performance and body composition of pigs.

High ambient temperature and air humidity are the most important climatic factors that jeopardize pig production. Multiple strategies can be applied for pigs under heat stress, including recent research to improve understanding the use of nutrition to attenuate the impact of heat stress. Heat stress impairs digestion, absorption, and amino acid metabolism with changes in amino acid requirements. Updates on the nutritional assessment strategies by differing the diets by protein and amino acid content (protein-bound or feed-grade) seem to be efficient tools for pork producers as amino acids play a functional role in challenged pigs apart from the beneficial effects on performance.

A different transcriptional landscape sheds light on Russian sturgeon (Acipenser gueldenstaedtii) mechanisms to cope with bacterial infection and chronic heat stress.

Sturgeons are chondrostean fish of high economic value and critically endangered due to anthropogenic activities, which has led to sturgeon aquaculture development. Russian sturgeon (Acipenser gueldenstaedtii), the second most important species reared for caviar, is successfully farmed in subtropical countries, including Uruguay. However, during the Uruguayan summer, sturgeons face intolerable warmer temperatures that weaken their defences and favour infections by opportunistic pathogens, increasing fish mortality and farm economic losses. Since innate immunity is paramount in fish, for which the liver plays a key role, we used deep RNA sequencing to analyse differentially expressed genes in the liver of Russian sturgeons exposed to chronic heat stress and challenged with Aeromonas hydrophila. We assembled 149.615 unigenes in the Russian sturgeon liver transcriptome and found that metabolism and immune defence pathways are among the top five biological processes taking place in the liver. Chronic heat stress provoked profound effects on liver biological functions, up-regulating genes related to protein folding, heat shock response and lipid and protein metabolism to meet energy demands for coping with heat stress. Besides, long-term exposure to heat stress led to cell damage triggering liver inflammation and diminishing liver ability to mount an innate response to A. hydrophila challenge. Accordingly, the reprogramming of liver metabolism over an extended period had detrimental effects on fish health, resulting in weight loss and mortality, with the latter increasing after A. hydrophila challenge. To our knowledge, this is the first transcriptomic study describing how chronic heat-stressed sturgeons respond to a bacterial challenge, suggesting that liver metabolism alterations have a negative impact on the innate anti-bacterial response.

Effects of free and dipeptide forms of methionine supplementation on oxidative metabolism of broilers under high temperature.

Our previous studies have shown that methionine supplementation could help to attenuate the effects of heat stress on the metabolism of broiler chickens. Here we investigated for the first time the effects of methionine supplementation in the form of DL-methionyl-DL-methionine on broilers subjected to heat stress during the growth phase. Broilers were divided into two groups; one group was reared under thermoneutral conditions and the other under continuous heat stress (30 ±â€¯1 °C, 60% relative humidity). Both groups were subdivided into three dietary treatments: a methionine-deficient (MD) diet, a diet supplemented with free methionine (DL-M), and a diet supplemented with methionine dipeptide (DL-MM). Broilers raised under chronic heat stress had lower feed intake and weight gain than broilers raised under thermoneutral conditions (P < 0.05). There were no differences in animal performance between methionine-supplemented diets (DL-M and DL-MM). Heat-stressed birds had significantly higher heterophil/lymphocyte (H/L) ratio than thermoneutral birds. Under heat stress, broilers fed DL-M and DL-MM diets had lower H/L ratio than birds fed the MD diet. Higher concentrations of carbonylated proteins and lower concentration of reduced glutathione were observed in broilers raised under heat stress. In comparing heat-stressed broilers, we found that birds fed the DL-M diet had lower concentrations of thiobarbituric acid-reactive substances and carbonylated proteins than those fed the MD diet (P < 0.05). Higher expression of glutathione peroxidase (GPX) and glutathione synthetase (GSS) genes was observed in heat-stressed broilers (P < 0.05). Under heat stress, the MD diet increased GPX expression compared with other diets. Under thermoneutral conditions, the DL-M diet resulted in the highest GSS expression. There was a negative correlation between DNA methylation and GPX and GSS expression. Our results showed that supplementation of broiler diets with free methionine or methionine dipeptide may help attenuate the effects of heat stress through enhanced activation of genes related to the glutathione antioxidant system. Methionine effects were found for gene regulation, gene expression, and post-translational processing.

Modulating offspring responses: concerted effects of stress and immunogenic challenge in the parental generation.

The perception, processing and response to environmental challenges involves the activation of the immuno-neuroendocrine (INE) interplay. Concerted environmental challenges might induce trade-off when resource allocation to one trait occurs at the expense of another, also producing potential transgenerational effects in the offspring. We evaluated whether concerted challenges, in the form of an immune inoculum against inactivated Salmonella enteritidis (immune challenge, ICH) and a chronic heat stress (CHS) exposure on adult Japanese quail, modulate the INE responses of the parental generation and their offspring. Adults were inoculated and later exposed to a CHS along nine consecutive days. For the last 5 days of the CHS, eggs were collected for incubation. Chicks were identified according to their parental treatments and remained undisturbed. Induced inflammatory response, heterophil/lymphocyte (H/L) ratio and specific humoral response against sheep red blood cells (SRBC) were evaluated in both generations. Regardless of the ICH, stressed adults showed a reduced inflammatory response and an elevated H/L ratio compared with controls. In offspring, the inflammatory response was elevated and the specific SRBC antibody titres were diminished in those chicks prenatally exposed to CHS, regardless of the ICH. No differences were found in the H/L ratio of the offspring. Together, our results suggest that CHS exposure influences the INE interplay of adult quail, establishing trade-offs within their immune system. Moreover, CHS not only affected parental INE responses but also modulated their offspring INE responses, probably affecting their potential to respond to future challenges. The adaptability of the developmental programming of offspring would depend on the environment encountered.

Effects of Dietary Supplementation of Fumaric Acid on Growth Performance, Blood Hematological and Biochemical Profile of Broiler Chickens Exposed to Chronic Heat Stress

The aim of this study was to investigate the effects of dietary fumaric acid (FA) on the growth performance, hematological parameters and serum biochemistry in broilers under heat stress (HS) condition. A total number of 200 broilers were allocated to five treatments including the thermoneutral group, which was reared at standard ambient temperature and fed basal diet (CONT) and four heat-stressed groups, which were held at 32 ± 1ºC aged from 22 to 42 days and fed the basal diet supplemented with 0, 5, 10 and 15 g/kg FA, respectively. On day 42, body weight and feed intake were measured and feed conversion ratio was calculated. Blood was collected for the estimation of hematological and biochemical parameters. HS impaired the growth performance, but the addition of FA made a higher final body weight, average daily gain and European broiler index in comparison with HS group. The hemoglobin concentration and packed cell volume of HS broilers were significantly lower than those of the CONT birds. Birds administered 5g/kg FA diet had significantly (p 0.05) higher erythrocyte counts and hemoglobin concentration compared with birds in the HS group fed basal diet. Also, the HS group exhibited significant increase in glucose, triglyceride, total cholesterol, low-density lipoprotein cholesterol and creatinine when compared with CONT group. Dietary 5g/kg FA increased the serum total protein, albumin, globulin, total cholesterol, high-density lipoprotein cholesterol in comparison with the HS group. In conclusion, this study demonstrated the importance of using FA as feed additives to improve the growth performance of heat-stressed broilers through haematological and biochemical regulation.

Effects of Dietary Supplementation of Fumaric Acid on Growth Performance, Blood Hematological and Biochemical Profile of Broiler Chickens Exposed to Chronic Heat Stress

The aim of this study was to investigate the effects of dietary fumaric acid (FA) on the growth performance, hematological parameters and serum biochemistry in broilers under heat stress (HS) condition. A total number of 200 broilers were allocated to five treatments including the thermoneutral group, which was reared at standard ambient temperature and fed basal diet (CONT) and four heat-stressed groups, which were held at 32 ± 1ºC aged from 22 to 42 days and fed the basal diet supplemented with 0, 5, 10 and 15 g/kg FA, respectively. On day 42, body weight and feed intake were measured and feed conversion ratio was calculated. Blood was collected for the estimation of hematological and biochemical parameters. HS impaired the growth performance, but the addition of FA made a higher final body weight, average daily gain and European broiler index in comparison with HS group. The hemoglobin concentration and packed cell volume of HS broilers were significantly lower than those of the CONT birds. Birds administered 5g/kg FA diet had significantly (p 0.05) higher erythrocyte counts and hemoglobin concentration compared with birds in the HS group fed basal diet. Also, the HS group exhibited significant increase in glucose, triglyceride, total cholesterol, low-density lipoprotein cholesterol and creatinine when compared with CONT group. Dietary 5g/kg FA increased the serum total protein, albumin, globulin, total cholesterol, high-density lipoprotein cholesterol in comparison with the HS group. In conclusion, this study demonstrated the importance of using FA as feed additives to improve the growth performance of heat-stressed broilers through haematological and biochemical regulation.(AU)