Effect of chlorogenic acid on intestinal inflammation, antioxidant status, and microbial community of young hens challenged with acute heat stress.

Heat stress in poultry is deleterious to productive performance. Chlorogenic acid (CGA) exerts antibacterial, anti-inflammatory, and antioxidant properties. This study was conducted to evaluate the effects of dietary supplemental CGA on the intestinal health and cecal microbiota composition of young hens challenged with acute heat stress. 100-day-old Hy-line brown pullets were randomly divided into four groups. The control group (C) and heat stress group (HS) received a basal diet. HS + CGA300 group and HS + CGA600 group received a basal diet supplemented with 300- and 600-mg/kg CGA, respectively, for 2 weeks before heat stress exposure. Pullets of HS, HS + CGA300 , and HS + CGA600 group were exposed to 38°C for 4 h while the control group was maintained at 25°C. In this study, dietary CGA supplementation had effect on mitigate the decreased T-AOC and T-SOD activities and the increasing of IL-1ß and TNFα induced by acute heat stress. Dietary supplementation with 600 mg/kg CGA had better effect on increasing the relative abundance of beneficial bacterial genera, such as Rikenellaceae RC9_gut_group, Ruminococcaceae UCG-005, and Christensenellaceae R-7_group, and deceasing bacteria genera involved in inflammation, such as Sutterella species. Therefore, CGA can ameliorate acute heat stress damage through suppressing inflammation and improved antioxidant capacity and cecal microbiota composition.

Nutritional manipulation to combat heat stress in poultry - A comprehensive review.

Global warming and climate change adversely affect livestock and poultry production sectors under tropical and subtropical conditions. Heat stress is amongst the most significant stressors influencing poultry productivity in hot climate regions, causing substantial economic losses in poultry industry. These economic losses are speculated to increase in the coming years with the rise of global temperature. Moreover, modern poultry strains are more susceptible to high ambient temperature. Heat stress has negative effects on physiological response, growth performance and laying performance, which appeared in the form of reducing feed consumption, body weight gain, egg production, feed efficiency, meat quality, egg quality and immune response. Numerous practical procedures were used to ameliorate the negative impacts of increased temperature; among them the dietary manipulation, which gains a great concern in different regions around the world. These nutritional manipulations are feed additives (natural antioxidants, minerals, electrolytes, phytobiotics, probiotics, fat, and protein), feed restriction, feed form, drinking cold water and others. However, in the large scale of poultry industry, only a few of these strategies are commonly used. The current review article deliberates the different practical applications of useful nutritional manipulations to mitigate the heat load in poultry. The documented information will be useful to poultry producers to improve the general health status and productivity of heat-stressed birds via enhancing stress tolerance, oxidative status and immune response, and thereby provide recommendations to minimize production losses due to heat stress in particular under the growing global warming crisis.

Heat stress influence the microbiota and organic acids concentration in beef cattle rumen.

The objective of this study was to evaluate the effect of heat stress on meta-taxonomic and metabolic profiles of prokaryotes in beef cattle rumen. Six pure-breed Nellore heifers with ruminal cannulas were used in the study. Six treatments were tested in a 6 × 6 Latin Square with six periods of 21days. The treatments were evaluated in a 2 × 2 + 2 factorial arrangement, consisting of 4 combinations: two temperatures conditions (thermoneutral, TN: 24 °C; and heat stress, HS: 34 °C) and two dietary energy concentration [low-energy (37% non-fibrous carbohydrates - NFC, 12 Mcal of metabolizable energy per kg of dry matter) or high-energy concentration (50.5% NFC, 18.49 Mcal of metabolizable energy per kg of dry matter)] plus two additional treatments with animals maintained in TN conditions but with your intake restricted (TN-RI) to the same of the heifers in HS with the two dietary energy concentration. The meta-genome was sequenced by MiSeq Sequencing System platform, and the DNA sequences were analysed using Geneious 10.2.3 software. The metabolic profile was evaluated by liquid and gas chromatography. Animals under HS presented lower (P = 0.04) prokaryote richness than animals under TN conditions. The genera Flavonifractor (1.4%), Treponema (0.6%) and Ruminococcus (0.9%) showed the lowest (P < 0.04) and Carnobacterium (7.7%) the highest (P = 0.02) relative abundance when the animals were submitted to HS, in relation to animals in TN. A total of 49 different metabolites were identified in the ruminal samples. The concentration of isobutyric acid (4.32 mM) was highest in bovine rumen under HS conditions. Heat stress influenced the microbiota and concentration of some organic acids in beef cattle rumen. There was a reduction in the richness of rumen in cattle under heat stress, but the diversity of prokaryotes was not affected.

Influence of aerobic fitness on gastrointestinal barrier integrity and microbial translocation following a fixed-intensity military exertional heat stress test.

PURPOSE: Exertional-heat stress adversely disrupts gastrointestinal (GI) barrier integrity, whereby subsequent microbial translocation (MT) can result in potentially serious health consequences. To date, the influence of aerobic fitness on GI barrier integrity and MT following exertional-heat stress is poorly characterised. METHOD: Ten untrained (UT; VO2max = 45 ± 3 ml·kg-1·min-1) and ten highly trained (HT; VO2max = 64 ± 4 ml·kg-1·min-1) males completed an ecologically valid (military) 80-min fixed-intensity exertional-heat stress test (EHST). Venous blood was drawn immediately pre- and post-EHST. GI barrier integrity was assessed using the serum dual-sugar absorption test (DSAT) and plasma Intestinal Fatty-Acid Binding Protein (I-FABP). MT was assessed using plasma Bacteroides/total 16S DNA. RESULTS: UT experienced greater thermoregulatory, cardiovascular and perceptual strain (p < 0.05) than HT during the EHST. Serum DSAT responses were similar between the two groups (p = 0.59), although Δ I-FABP was greater (p = 0.04) in the UT (1.14 ± 1.36 ng·ml-1) versus HT (0.20 ± 0.29 ng·ml-1) group. Bacteroides/Total 16S DNA ratio was unchanged (Δ; -0.04 ± 0.18) following the EHST in the HT group, but increased (Δ; 0.19 ± 0.25) in the UT group (p = 0.05). Weekly aerobic training hours had a weak, negative correlation with Δ I-FABP and Bacteroides/total 16S DNA responses. CONCLUSION: When exercising at the same absolute workload, UT individuals are more susceptible to small intestinal epithelial injury and MT than HT individuals. These responses appear partially attributable to greater thermoregulatory, cardiovascular, and perceptual strain.

The effect of heat stress on intestinal integrity and Salmonella invasion in broiler birds.

The intestinal mucosa works as a barrier to protect the internal environment of the animal from bacteria and bacterial toxins found in the gut lumen. Heat stress may harm this function. Therefore, we designed the current experiment to investigate the effect of heat stress on intestinal integrity, physiological and immunological responses and Salmonella invasion in broiler chickens. At 26 days of age, 72 birds were randomly distributed into 3 treatments, with 8 replicates per treatment and 3 birds per replicate. The three treatments were control treatment; kept at thermoneutral environmental conditions (20 ± 2°C), chronic heat stress treatment (exposed to 30 ± 2°C; 24h/day) and acute heat stress treatment (exposed to 35 ±2°C from 09:00 to 13:00 and kept at 20 ± 1°C from 13:00 to 09:00). The heat stress exposure was conducted for 10 successive days. Compared with the control treatment, birds subject to chronic and acute heat stress had reduced (P < 0.05) body weight and body gain and increased (P < 0.05) feed conversion ratio. However, feed intake and mortality rate were only increased (P < 0.05) in the acute heat stress treatment. Rectal temperature and Δ rectal temperature (°C/h) increased (P < 0.05) sharply during the first 2 days of exposure followed by gradual decreases until a plateau was achieved. Heat-stressed birds had increased (P < 0.05) serum concentrations of corticosterone, endotoxin lipopolysaccharide and the systemic inflammatory cytokine: TNF-α and IL-2, as well as a higher (P < 0.05) prevalence of Salmonella spp. in meat and livers, as compared with control treatment. It can be concluded that heat stress impaired intestinal integrity which resulted in increased intestinal permeability to endotoxin, translocation of intestinal pathogens (Salmonella spp.) and serum inflammatory cytokines. Therefore, avoiding thermal dysfunction of intestinal barrier is a significant factor in maintaining welfare, immune status and meat safety of broiler birds.

Metabolomics revealed diurnal heat stress and zinc supplementation-induced changes in amino acid, lipid, and microbial metabolism.

Heat stress (HS) dramatically disrupts the events in energy and nutrient metabolism, many of which requires zinc (Zn) as a cofactor. In this study, metabolic effects of HS and Zn supplementation were evaluated by examining growth performance, blood chemistry, and metabolomes of crossbred gilts fed with ZnNeg (no Zn supplementation), ZnIO (120 ppm ZnSO4), or ZnAA (60 ppm ZnSO4 + 60 ppm zinc amino acid complex) diets under diurnal HS or thermal-neutral (TN) condition. The results showed that growth performance was reduced by HS but not by Zn supplementation. Among measured serum biochemicals, HS was found to increase creatinine but decrease blood urea nitrogen (BUN) level. Metabolomic analysis indicated that HS greatly affected diverse metabolites associated with amino acid, lipid, and microbial metabolism, including urea cycle metabolites, essential amino acids, phospholipids, medium-chain dicarboxylic acids, fatty acid amides, and secondary bile acids. More importantly, many changes in these metabolite markers were correlated with both acute and adaptive responses to HS. Relative to HS-induced metabolic effects, Zn supplementation-associated effects were much more limited. A prominent observation was that ZnIO diet, potentially through its influences on microbial metabolism, yielded different responses to HS compared with two other diets, which included higher levels of short-chain fatty acids (SCFAs) in cecal fluid and higher levels of lysine in the liver and feces. Overall, comprehensive metabolomic analysis identified novel metabolite markers associated with HS and Zn supplementation, which could guide further investigation on the mechanisms of these metabolic effects.

Identification of diagnostic biomarkers and metabolic pathway shifts of heat-stressed lactating dairy cows.

Controlling heat stress (HS) is a global challenge for the dairy industry. However, simple and reliable biomarkers that aid the diagnoses of HS-induced metabolic disorders have not yet been identified. In this work, an integrated metabolomic and lipidomic approach using (1)H nuclear magnetic resonance and ultra-fast LC-MS was employed to investigate the discrimination of plasma metabolic profiles between HS-free and HS lactating dairy cows. Targeted detection using LC-MS in multiple reaction monitoring mode was used to verify the reliability of the metabolites as biomarker candidates. Overall, 41 metabolites were identified as candidates for lactating dairy cows exposed to HS, among which 13 metabolites, including trimethylamine, glucose, lactate, betaine, creatine, pyruvate, acetoacetate, acetone, ß-hydroxybutyrate, C16 sphinganine, lysophosphatidylcholine (18:0), phosphatidylcholine (16:0/14:0), and arachidonic acid, had high sensitivity and specificity in diagnosing HS status, and are likely to be the potential biomarkers of HS dairy cows. All of these potentially diagnostic biomarkers were involved in carbohydrate, amino acid, lipid, or gut microbiome-derived metabolism, indicating that HS affected the metabolic pathways in lactating dairy cows. Further research is warranted to evaluate these biomarkers in practical applications and to elucidate the physiological mechanisms of HS-induced metabolic disorders. BIOLOGICAL SIGNIFICANCE: Heat stress (HS) annually causes huge losses to global dairy industry, including animal performance decrease, metabolic disorder and health problem. So far, physiological mechanisms underlying HS of dairy cows still remain elusive. To our best knowledge, this is the first attempt to elucidate the HS-induced metabolic disorders of dairy cows using integrated (1)H NMR and LC-MS-based metabolic study. The results not only provided potential diagnostic biomarkers for HS lactating dairy cows, but also significantly explored the related physiological mechanisms of metabolic pathway shifts induced by HS environment. This work offers comprehensive insights into the global metabolic alterations of dairy cows exposed to HS and provides a new perspective for further study.

Oral administration of Bacillus subtilis strain BSB3 can prevent heat stress-related adverse effects in rats.

AIMS: To determine the efficacy of Bacillus subtilis strain in prevention of heat stress-related complications in rats. METHODS AND RESULTS: Male Sprague-Dawley rats weighing 250-300 g were treated by oral gavage with B. subtilis BSB3 strain or PBS twice a day for 2 days. Half of the rats of each group were exposed to heat stress (45°C, relative humidity 55% for 25 min), while the remaining rats were placed at identical conditions but at 25°C. Bacterial translocation, histological changes in the intestine, cytokines profile, serum lipopolysaccharide level (LPS), as well as vesiculation of erythrocytes were analysed and compared between groups. Adverse effects of heat stress (morphological changes in intestine, bacterial translocation, elevated levels of LPS and IL-10, increased vesiculation of erythrocytes) were observed only in rats not protected with B. subtilis strain and exposed to heat. All registered parameters in rats pretreated with bacilli and exposed to heat were similar to control groups. CONCLUSIONS: Bacillus subtilis BS3B strain was effective in the prevention of complications related to heat stress in rats. SIGNIFICANCE AND IMPACT OF THE STUDY: This work will contribute towards better understanding of probiotics' mechanisms of action and will bring new approaches to characterize and use of beneficial bacteria.

Effects of long-term heat stress in an experimental model of avian necrotic enteritis.

Stressful conditions are predisposing factors for disease development. Heat stress is one of the most important stressors in poultry production. The reemergence of some previously controlled diseases [e.g., avian necrotic enteritis (NE)] has been extensively reported. The combination of bacterial infection and certain environmental factors have been reported to trigger the disease. The aim of this study was to analyze the effects of long-term heat stress (35 ± 1°C) on the development of NE in broiler chickens. For this purpose, 60 male broiler chickens were divided into the following 6 groups: control group (C), heat stressed control group (C/HS35), thioglycolate group (T), thioglycolate heat-stressed group (T/HS35), infected group (I), and infected heat-stressed group (I/HS35). The poultry of groups I and I/HS35 were experimentally infected with Clostridium perfringens via their feed from 15 to 21 d of life. Heat stress (35 ± 1°C) was constantly applied to the birds of the stressed groups from 14 to 21 d of life. The infected and heat-stressed broiler chickens presented a trend toward a decrease in gross lesion scores and significantly lower microscopic scores of necrosis in the duodenum and jejunum (P < 0.05), lower fusion of villi in the duodenum (P < 0.05), and lower congestion scores in the jejunum and ileum (P < 0.05) in relation to infected and non-heat-stressed chickens. Broilers of I/HS35 group also exhibited small number of heterophils in the duodenum and jejunum compared with those of the I group (P < 0.05). Furthermore, the duodenum and jejunum of infected and heat-stressed broilers showed lower number of clostridia on the intestinal mucosa (P < 0.05). Data were discussed in light of a heat stress induced reduction on intestinal inflammation via a decrease in heterophil migration to the intestinal mucosa, which in turn might have reduced tissue damage during inflammation, hence preventing the development of a more severe form of NE.

Haemodynamics of lambs grazing perennial ryegrass (Lolium perenne L.) either infected with AR6 novel, wild-type endophyte, or endophyte-free.

AIM: To compare vasoconstriction of the auricular artery and characteristics of blood flow in the carotid arteries between lambs grazing perennial ryegrass that was either infected with the AR6 novel endophyte (AR6), wild-type endophyte or endophyte-free. METHODS: Sixteen Coopworth lambs, mean 23.7 (SD 1.8) kg, were randomly assigned to graze three, 0.10-ha pastures of perennial ryegrass (cultivar Extreme) located in Lincoln, New Zealand, that were infected with either the AR6 novel endophyte (n=5) or wild-type endophyte (n=6) or were endophyte-free (n=5), for 18 days until 16 March 2009. Lambs on AR6 pasture were then switched to endophyte-free pasture, and those on endophyte-free were switched to AR6 pasture, for 18 days. Lambs continued grazing the wild-type ryegrass during both phases of the study. Colour Doppler ultrasonography was used to monitor cross-sectional area of lumina in the auricular and carotid arteries as measurements of vasoconstriction, and to measure pulsatility indices, heart rate, systolic and diastolic velocities, and mean velocity in the carotid artery. Urine was sampled once during each phase, to measure the concentration of urinary alkaloids. A heat challenge (32°C) was imposed on the last day of the experiment, to determine treatment effects on rectal temperature and respiration rate. RESULTS: Vasoconstriction was detected in the auricular arteries of lambs grazing AR6 and wild-type pastures. Areas of lumina declined linearly over 9 days in lambs that were switched from endophyte-free to AR6 pastures (p<0.05), and areas increased linearly in lambs that were switched from AR6 to endophyte-free pasture (p<0.001). Resistance to blood flow in the carotid arteries decreased linearly in the 9 days after lambs were switched from AR6 to endophyte-free pasture(p<0.05), and tended to increase after lambs on endophyte-free pasture were switched to AR6 (p<0.10). The concentrations of urinary alkaloids decreased after switching lambs from AR6 to endophyte-free pasture, and increased after switching from endophyte-free to AR6 pasture (p<0.05). The concentrations of urinary alkaloids of lambs on wild-type pasture were similar between the conditioning and experimental phases. There were no treatment effects on rectal temperature and respiration rate during the heat challenge. CONCLUSIONS: These results indicated that management approaches are needed to overcome potential vulnerabilities to heat stress for sheep grazing AR6 perennial ryegrass. Furthermore, following grazing such pasture, lambs will need to graze endophyte-free ryegrass for >18 days, to completely clear ergot alkaloids from their vasculature, assuming that complete clearance can be achieved.

Glycinebetaine alleviates the inhibitory effect of moderate heat stress on the repair of photosystem II during photoinhibition.

Transformation with the bacterial gene codA for choline oxidase allows Synechococcus sp. PCC 7942 cells to accumulate glycinebetaine when choline is supplemented exogenously. First, we observed two types of protective effect of glycinebetaine against heat-induced inactivation of photosystem II (PSII) in darkness; the codA transgene shifted the temperature range of inactivation of the oxygen-evolving complex from 40-52 degrees C (with half inactivation at 46 degrees C) to 46-60 degrees C (with half inactivation at 54 degrees C) and that of the photochemical reaction center from 44-55 degrees C (with half inactivation at 51 degrees C) to 52-63 degrees C (with half inactivation at 58 degrees C). However, in light, PSII was more sensitive to heat stress; when moderate heat stress, such as 40 degrees C, was combined with light stress, PSII was rapidly inactivated, although these stresses, when applied separately, did not inactivate either the oxygen-evolving complex or the photochemical reaction center. Further our studies demonstrated that the moderate heat stress inhibited the repair of PSII during photoinhibition at the site of synthesis de novo of the D1 protein but did not accelerate the photodamage directly. The codA transgene and, thus, the accumulation of glycinebetaine alleviated such an inhibitory effect of moderate heat stress on the repair of PSII by accelerating the synthesis of the D1 protein. We propose a hypothetical scheme for the cyanobacterial photosynthesis that moderate heat stress inhibits the translation machinery and glycinebetaine protects it against the heat-induced inactivation.

Effects of two probiotic Lactobacillus strains on jejunal and cecal microbiota of broiler chicken under acute heat stress condition as revealed by molecular analysis of 16S rRNA genes.

We examined the effects of probiotic Lactobacillus strains of Lactobacillus agilis JCM 1048 and Lactobacillus salivarius subsp. salicinius JCM 1230 on jejunal and cecal microbiota of broiler chicken under heat stress condition using terminal restriction fragment length polymorphism (T-RFLP) analysis. The jejunal bacterial community was limited to a few bacterial groups, mostly Lactobacillus spp. A relatively abundant and higher prevalence of Lactobacillus spp. were observed in the jejunal and cecal microbiota of the probiotic chickens compared with those of the control chickens under heat stress condition. In general, the probiotic strains did not significantly affect the abundance of L. agilis and L. salivarius in chicken intestine but clearly contributed to increasing their prevalence in the probiotic chickens. The probiotic Lactobacillus strains enriched the diversity of Lactobacillus flora in chicken jejunum and cecum by increasing the abundance and prevalence of Lactobacillus spp. inhabiting the intestine. The richness of Lactobacillus species tended to be similar among the jejunal and cecal microbiota. The bacterial community of cecum was complex and age-dependent. The major components of the cecal microbiota were clostridia and lactobacilli. The Clostridium subcluster XIVa was the most predominant group in chicken cecum. Probiotic Lactobacillus strains restored the microbial balance and maintained the natural stability of indigenous bacterial microbiota following heat stress-induced changes.