Effect of lairage time prior to slaughter on stress in pigs: a path analysis.

BACKGROUND: Pre-slaughter process during transportation, handling, and lairage causes stress in pigs, affecting animal welfare and meat quality. Therefore, lairage factors are important for relieving stress. A total of 24 LYD (Landrace × Yorkshire × Duroc) barrows were used to investigate the effect of 6 and 20 h lairage time (LT) on cortisol, serotonin, and catecholamine in blood and physiological factors in muscle, and to verify the causal relationship between these factors. RESULTS: The results revealed that cortisol was increased (0.064 ± 0.007 µg/ml), and epinephrine (0.020 ± 0.002 µg/ml) and norepinephrine (1.518 ± 0.071 µg/ml) were lower at a LT of 20 h than those at 6 h, and there was no significant effect on the muscle and carcass characteristic factors. In addition, cortisol and norepinephrine showed a negative correlation (r = -50,346, p = 0.0121), epinephrine and glycogen had a positive correlation (r = 0.4417, p = 0.0307), and serotonin and heat shock protein 70 (HSP70) were positively correlated (r = 0.4715, p = 0.0200). Path analysis indicated that the increase in LT had a direct effect on cortisol, epinephrine, and norepinephrine, and an indirect effect on muscle glycogen. CONCLUSION: This study confirmed the effect of the increase in LT from 6 to 20 h in the lairage room on the stress response of pigs. These findings support the legal requirements that advocate for shorter lairage times, in alignment with enhanced animal welfare standards.

COPA3 peptide supplementation alleviates the heat stress of chicken fibroblasts.

Heat stress inhibits cellular proliferation and differentiation through the production of reactive oxygen species. Under stress conditions, antioxidant drugs promote stable cellular function by reducing the stress level. We sought to demonstrate 9-mer disulfide dimer peptide (COPA3) supplementation stabilizes fibroblast proliferation and differentiation even under heat stress conditions. In our study, fibroblasts were assigned to two different groups based on the temperature, like 38°C group presented as Control - and 43°C group presented as Heat Stress-. Each group was subdivided into two groups depending upon COPA3 treatment, like 38°C + COPA3 group symbolized Control+ and the 43°C + COPA3 group symbolized as Heat Stress+. Heat stress was observed to decrease the fibroblast viability and function and resulted in alterations in the fibroblast shape and cytoskeleton structure. In contrast, COPA3 stabilized the fibroblast viability, shape, and function. Moreover, heat stress and COPA3 were found to have opposite actions with respect to energy production, which facilitates the stabilization of cellular functions by increasing the heat tolerance capacity. The gene expression levels of antioxidant and heat shock proteins were higher after heat stress. Additionally, heat stress promotes the mitogen-activated protein kinase/ extracellular signal-regulated kinase-nuclear factor erythroid 2-related factor 2 (MAPK/ERK-Nrf2). COPA3 maintained the MAPK/ERK-Nrf2 gene expressions that promote stable fibroblast proliferation, and differentiation as well as suppress apoptosis. These findings suggest that COPA3 supplementation increases the heat tolerance capacity, viability, and functional activity of fibroblasts.

HSP expression depends on its molecular construction and different organs of the chicken: a meta-analysis.

Heat shock proteins (HSPs) expression protect the cell from stress, this expression varies on tissue and stress level. Here, we investigated the structure and functional expression of HSPs in different chicken organs using meta-analysis. A total of 1253 studies were collected from three different electronic databases from January 1, 2015 to February 1, 2022. Of these studies, 28 were selected based on the specific criteria for this meta-analysis. The results for the expression of HSPs and the comparative expression of HSPs (HSP90, HSP70, and HSP60) in different chicken organs (brain, heart, liver, muscle, and intestine) were analyzed using the odds ratio or the random-effects model (REM) at a confidence interval (CI) of 95%. Compared to the thermoneutral groups, heat stress groups exhibited a significant (P < 0.01) change in their HSP70 expression in the chicken liver (8 trials: REM = 1.41, 95% CI: 0.41, 4.82). The expression of different HSPs in various chicken organs varied and the different organs were categorized according to their expression levels. HSP expression differed among the heart, liver, and muscle of chickens. HSPs expression level depends on the structure and molecular weight of the HSPs, as well as the type of tissue.

Transcriptome-wide analysis reveals gluten-induced suppression of small intestine development in young chickens.

Wheat gluten is an increasingly common ingredient in poultry diets but its impact on the small intestine in chicken is not fully understood. This study aimed to identify effects of high-gluten diets on chicken small intestines and the variation of their associated transcriptional responses by age. A total of 120 broilers (Ross Strain) were used to perform two animal experiments consisting of two gluten inclusion levels (0% or 25%) by bird's age (1 week or 4 weeks). Transcriptomics and histochemical techniques were employed to study the effect of gluten on their duodenal mucosa using randomly selected 12 broilers (3 chicks per group). A reduction in feed intake and body weight gain was found in the broilers fed a high-gluten containing diet at both ages. Histochemical photomicrographs showed a reduced villus height to crypt depth ratio in the duodenum of gluten-fed broilers at 1 week. We found mainly a significant effect on the gene expression of duodenal mucosa in gluten-fed broilers at 1 week (289 differentially expressed genes [DEGs]). Pathway analyses revealed that the significant DEGs were mainly involved in ribosome, oxidative phosphorylation, and peroxisome proliferator-activated receptor (PPAR) signaling pathways. These pathways are involved in ribosome protein biogenesis, oxidative phosphorylation and fatty acid metabolism, respectively. Our results suggest a pattern of differential gene expression in these pathways that can be linked to chronic inflammation, suppression of cell proliferation, cell cycle arrest and apoptosis. And via such a mode of action, high-gluten inclusion levels in poultry diets could lead to the observed retardation of villi development in the duodenal mucosa of young broiler chicken.

Meta-Analysis and Systematic Review of the Thermal Stress Response: Gallus gallus domesticus Show Low Immune Responses During Heat Stress.

Heat stress, which affects broiler growth performance and immunity, is a major concern in the poultry industry. This meta-analysis aimed to demonstrate the significant effect of heat stress on broiler mass gain and immunoglobulin levels, which regulates the mortality rate of broilers. A total of 2,585 studies were downloaded from PubMed, Web of Science, and Google Scholar from January 1, 2015, to September 1, 2021. Eventually, 28 studies were selected based on specific criteria. The results for body mass gain, total mass of immune organs (thymus, spleen, and bursa of Fabricius), immunoglobulin (IgA, IgG, and IgM) levels, and mortality rate were analyzed using odds ratio or the random-effects model (REM) at a confidence interval (CI) of 95%. Compared to the control, heat stress significantly decreased body mass gain (10 trials: REM = 1.35, 95% CI: 1.21, 1.50). Compared to that in the control, heat stress significantly increased immunoglobulin levels: IgA (7 trials: REM = 1.69, 95% CI: 0.90, 3.16), IgG (6 trials: REM = 1.24, 95% CI: 0.85, 1.81), IgM (8 trials: REM = 0.69, 95% CI: 0.44, 1.08), and heat stress also increased the broiler mortality rate (6 trials: REM = 0.06, 95% CI: 0.01, 0.27). However, there were no significant changes in the immune organs between the control and heat-stressed groups. In conclusion, heat stress remarkably alters the mass gain and immunoglobulin levels of broilers, which may be a cause of the high mortality rate.

Cortisol differentially affects the viability and myogenesis of mono- and co-cultured porcine gluteal muscles satellite cells and fibroblasts.

Cortisol is a ubiquitously expressed stress hormone. In this study, we investigated the effects of exogenous cortisol on porcine gluteal muscles primary cultured satellite cells and fibroblasts. Satellite cells and fibroblasts were mono-or co-cultured, and cells in each type of culture were categorized into the control and cortisol-treated (treatment) groups. We selected 28 µmol mL-1 cortisol for treatment based on their efficacy. Cortisol treatment reduced viability of monocultured satellite cells and fibroblasts. In both monocultured and co-cultured cells, the nucleus in the treatment group was damaged than that control group. Moreover, the total cell cycle duration was shorter in the treatment group than the control group. PAX-7 expression was upregulated in the control group of co-cultured satellite cells and fibroblasts than those remaining groups. Moreover, MyoD expression was downregulated in the cortisol treated group of both mono-and co-cultured satellite cells compared with that in the control group. In co-cultured fibroblasts, MyoD and MyoG expression was upregulated than those remaining groups. The Cyto-C expression was upregulated in the treatment group compared to the control mono-and co-cultured both cells. These results suggest that the selected experimental dose of cortisol reduced cell viability and myogenesis-related gene expression in the monoculture compared to that in the co-culture of satellite cells and fibroblasts.

Altered relationship between gluconeogenesis and immunity in broilers exposed to heat stress for different durations.

This study determined the relationship between inflammation and gluconeogenesis level in broilers in different durations of heat stress. A total of 240 Ross 308 broilers were offered control and heat stress temperature from 21 to 35 d post-hatch, each experimental group had 8 replications, and each replication obtained 15 broilers. The temperature in the control (Ctrl) group and heat stress group were maintained at 24 ± 1°C and 34 ± 1°C, respectively throughout the experimental period. Based on the duration of heat stress, the heat stress group was divided into 2 subgroups, like, 7-d heat stress (28-day-old broiler) designated ST group and 14-d heat stress (35-day-old broiler) designated the LT group. The ad libitum commercial feed and fresh water were provided to all experimental broilers during the experiment. The growth performance of experimental broilers was calculated at 35 d. However, the liver and blood samples were collected from the Ctrl group in 21 d, as well as these samples were collected from the heat stress ST and LT groups in 28-d and 35-d, respectively. Obvious gene expression of immunity, gluconeogenesis, glycogenolysis, and glycogenesis, as well as glucose-6-phosphate dehydrogenase and adenosine triphosphate was determined in the liver sample. The blood glucose concentration and histopathology of the liver was also examined in the different grouped broilers. Body weight, weight gain, and feed intake significantly decreased in the 35-d heat stress group than the Ctrl group. However, the feed conversion ratio increased at the 35-d heat stress group than the Ctrl group. The amount of glucose-6-phosphate dehydrogenase was significantly higher in ST and LT groups than Ctrl, whereas the blood glucose level was downregulated in the LT group. The amount of adenosine triphosphate was significantly decreased in the LT group than the Ctrl and ST groups. Heat stress acts as an impediment to the general relation between gluconeogenesis and immunity, as well as changes cellular structure. This experiment contributed to the establishment of a relationship between gluconeogenesis and immunity, which affects the growth performance of broilers during heat stress.

Early Heat Exposure Effects on Proteomic Changes of the Broiler Liver under Acute Heat Stress.

As environmental temperatures continue to rise, heat stress (HS) is having a negative effect on the livestock industry. In order to solve this problem, many studies have been conducted to reduce HS. Among them, early heat exposure has been suggested as a method for reducing HS in poultry. In this study, we analyzed proteomics and tried to identify the metabolic mechanisms of early heat exposure on acute HS. A total of 48 chicks were separated into three groups: CC (control groups raised at optimum temperature), CH (raised with CC but exposed acute HS at the 35th day), and HH (raised with CC but exposed early heat at the fifth day and acute HS at the 35th day). After the whole period, liver samples were collected for proteomic analysis. A total of 97 differentially expressed proteins were identified by acute HS. Of these, 62 proteins recovered their expression levels by early heat exposure. We used these 62 proteins to determine the protective effects of early heat exposure. Of the various protein-related terms, we focused on the oxidative phosphorylation, fatty acid metabolism, carbohydrate metabolism, and energy production metabolism. Our findings suggest the possibility of early heat exposure effects in acute HS that may be useful in breeding or management techniques for producing broilers with high heat resistance.

Early heat exposure effect on the heat shock proteins in broilers under acute heat stress.

The effects of early heat conditioning on the acute heat stress response in broilers were investigated via the growth performance, dopamine, serotonin, and corticosterone and the expression of heat shock proteins (HSP) and heat shock factors. One-day-old chicks (n = 144) were divided into 3 groups in a 35-d experiment (48 chicks per each group). Group 1 (C) was treated with an optimum temperature, group 2 (CH) was treated with 40°C ± 1°C on day 35 (5 h), and group 3 (HH) was treated with 40°C ± 1°C on day 5 (24 h) and day 35 (5 h). On day 7, the body weight gain was lower (P < 0.05) in HH than in C and CH. On day 35, the heat-treated groups (CH and HH) had lower weight gains than the C group (P < 0.05), whereas the feed conversion ratio was lower in HH (P < 0.05). Serum corticosterone was higher in CH than in C, but HH and C did not differ (P < 0.05). Liver HSP70 protein expression was higher in CH than HH and C (P < 0.05), which did not differ, and HSP40 protein expression was higher in CH than C (P < 0.05). These results suggest that early heat conditioning may reduce acute heat stress on broiler.

Modulatory effect of heat stress on viability of primary cultured chicken satellite cells and expression of heat shock proteins ex vivo.

Satellite cells promote muscle repairing and muscle growth. Thereby the intention of the present study was to investigate the beneficial effects of heat stress at different time intervals on chicken satellite cells' viability. Satellite cells were isolated from 1-day-old chicks and treated at two different temperatures (37 °C and 41 °C) for various time periods (6 h, 12 h, 24 h, 48 h, and 72 h). Both temperatures significantly increased cell viability after 24 h and 48 h. After 12 h, cell viability was significantly increased at 41 °C compared to 37 °C. However, more apoptotic cells were observed at end of the experiment of 41 °C compared to 37 °C. In addition, more live cells were found at early of experimental period at 41 °C than 37 °C. Additionally, protein and mRNA expression of HSP70, HP60 and HSP47 were significantly upregulated throughout the experimental period at temperature of 41 °C compared to those at 37 °C. These results indicate that cell viability and expression of heat stress related proteins/genes are induced by high temperature of 41 °C via heat stress pathway whereas activation of heat stress related proteins/genes are lower at 37 °C. Thus, 41 °C can trigger satellite cells' viability essential for better cell survival than 37 °C at early incubation time.

Chronic heat stress regulates the relation between heat shock protein and immunity in broiler small intestine.

Chronic heat stress is considered to decrease the immune functions which makes negative effect on broiler growth performance. Here, we investigated the relationship between chronic heat stress, growth performance, and immunity in the small intestine of broilers. The study included two groups (control and heat stressed group) with eight replications per group. Ten broilers of 20-day aged were allocated in each replication. On day 35, the treatment group was subdivided into two groups based on their body weights (heavy and low body weight). Although, there was only the control and treatment group on day 28. The growth performance decreased and expression of heat shock protein 70 (HSP70), HSP60, and HSP47 increased on days 28 and 35 in the chronic heat stress group as compared with those in the control group. The expression levels of HSPs were significantly higher in the low body weight group than in the control group. The genes HSP70 and HSP60 were significantly associated with pro- and anti-inflammatory cytokines in the small intestine of the broilers of the treatment group. Thus, HSP70 and HSP60 activated the adaptive immunity in the small intestines of the broilers from the treatment group to allow adaptation to chronic heat stress environment.

Direct exposure to mild heat stress stimulates cell viability and heat shock protein expression in primary cultured broiler fibroblasts.

Fibroblasts produce collagen which is mainly essential for repairing tissue damage and maintaining the structural integrity of tissues. However, studies have given scientific evidence about harmful effect of thermal manipulation in fibroblast. Therefore, the aim of this study was to determine the mild heat stress temperature which increased broiler fibroblast viability. The experiment was divided into two groups (37 °C and 41 °C), and each group was divided into five subgroups based on different incubation times (6 h, 12 h, 24 h, 48 h, and 72 h) with three replications. In experimental group (41 °C), fibroblast viability increased significantly in 12 h but decreased in 72 h compared with control (37 °C). At 41 °C, live cell increased significantly in 24 h and then declined in 48 h as well as 72 h than control. Moreover, the S phase lengthened in shorter incubation time of experimental group compared with control. Protein and mRNA (HSP70, HSP60, and HSP47) expressions were significantly higher at 41 °C compared with 37 °C, but at the end of the experiment, HSP expression level was higher in both groups. Finally, this study recommended 41 °C as a mild heat stress temperature for increasing broiler fibroblast viability.

Acute Heat Stress Induces the Differential Expression of Heat Shock Proteins in Different Sections of the Small Intestine of Chickens Based on Exposure Duration.

In this study, we examined the protein and gene expression of heat shock proteins (HSPs) in different sections of the small intestine of chickens. In total, 300 one-day-old Ross 308 broiler chicks were randomly allocated to the control and treatment groups. The treatment group was divided into four subgroups, according to the duration of acute heat exposure (3, 6, 12, and 24 h). The influence of heat stress on the protein and gene expression of HSP70, HSP60, and HSP47 in different sections of the small intestine of chickens was determined. The protein expression of HSP70 and HSP60 was significantly higher at 6 h in the duodenum and jejunum and 12 h in the ileum. The HSP47 protein expression was significantly higher at 3 h in the duodenum and ileum and at 6 h in the jejunum. The gene expression levels of HSP70, HSP60, and HSP47 were significantly higher at the 3 h treatment group than the control group in the duodenum, jejunum, and ileum. The glutamate pyruvate transaminase and glutamate oxaloacetate transaminase levels were significantly higher at 12 and 24 h in the serum of the blood. Acute heat stress affected the expression of intestinal proteins and genes in chickens, until the induction of heat tolerance.

Effects of In Ovo Supplementation with Nanonutrition (L-Arginine Conjugated with Ag NPs) on Muscle Growth, Immune Response and Heat Shock Proteins at Different Chicken Embryonic Development Stages.

The aim of the study was to analyze the in ovo injection of inorganic and organic synthesized silver nanoparticles (Ag NPs) using Brassica oleracea L. var. capitate F. rubra (BOL) conjugation with L-Arginine (L-Arg) on the immune, muscle growth, survivability and hatchability of broiler chickens. The conjugation of L-Arg (100 µg) with 1000 µg of Ag NPs synthesized by (BOL)-extract and L-Arg (100 µg) conjugated with 100 µg of Ag NPs inorganic synthesized were injected into fertile eggs at 8 d, 14 d and 18 d of incubation. Survival and hatching rate were significantly improved in the dose of L-Arg (100 µg) with 1000 µg (BOL-Ag NPs) and L-Arg (100 µg) with 100 µg (C-Ag NPs) on 14 d injection whereas it was decreased on 8 d or 18 d injection. Moreover, the protein expression of muscle development markers such as myogenin and myoD were significantly uprelated in 14 d of incubation whereas the heat shock proteins (HSPs), such as HSP-60 and HSP-70, were significantly upregulated in 18 d incubation. In addition, the liver function marker of serum glutamate oxaloacetate transaminase (SGOT) and serum glutamate pyruvate transaminase (SGPT) were significantly decreased and the immunoglobulin (IgM) levels were increased in a 14 d incubation period in serum at the same concentration.

Effects of white, yellow, and blue colored LEDs on milk production, milk composition, and physiological responses in dairy cattle.

Light emitting diode (LED) is more energy efficient than incandescent or fluorescent light. This study was to evaluate effects of different colored LEDs on milk production, milk composition, and physiology of Holstein cow. According to milk production and parity, cows (n = 186) were allotted to four treatments: control (natural daylight), white, yellow, and blue LED groups. Of these, 40 cows that had passed 57 day-in-milk were used. Yellow and blue LED groups demonstrated greater rates of decline in milk production than control and white LED groups. At the finish point, milk fat, protein, and lactose contents were the lowest in the blue LED group, whereas milk-urea-nitrogen levels were the highest in the yellow and blue LED groups. Extended exposure to blue LED light lowered antioxidant enzyme activity and insulin-like growth factor-1 levels. Prolactin concentrations were higher in the white and blue LED groups than in the control. Cortisol level was the highest in the blue LED group among the groups. Nonesterified fatty acid levels in the yellow and blue LED groups decreased to the greatest extent compared to the start point. These results suggest that blue LED light can decrease milk production and generate more stress than white and yellow LED lights.

Heat Treatment at an Early Age Has Effects on the Resistance to Chronic Heat Stress on Broilers.

This study was conducted to investigate the effects of early heat conditioning on growth performance, liver-specific enzymes (GOT and GPT), neuro-hormones (dopamine and serotonin), stress hormones (corticosterone), and the expression of HSPs (heat shock proteins), HSFs (heat shock factors), and pro-inflammatory cytokines under chronic high temperature. Broilers were raised with commercial feed and supplied with water ad libitum under conventional temperature. We separated the broilers into three groups: the control without any heat exposure (C), chronic heat-stressed group (CH), and early and chronic heat-stressed group (HH). At 5 days of age, the HH group was exposed to high temperatures (40 °C for 24 h), while the remaining groups were raised at a standard temperature. Between days 6 and 20, all three groups were kept under optimal temperature. From 21 to 35 days, the two heat-stressed groups (CH and HH) were exposed to 35 °C. Groups exposed to high temperature (CH and HH) showed significantly lower body weight and feed intake compared to the control. GOT and GPT were lower expressed in the CH and HH groups than the control group. In addition, the protein expressions of HSPs were down-regulated by chronic heat stress (CH and HH groups). The gene expressions of HSP60 and HSF3 were significantly down-regulated in the CH and HH groups, while HSP70 and HSP27 genes were up-regulated only in the HH group compared with the control group. The expression of pro-inflammatory cytokine genes was significantly up-regulated in the HH group compared with the control and CH groups. Thus, exposure of early Heat stress (HS) to broilers may affect the inflammatory response; however, early heat exposure did not have a positive effect on chronic HS of liver enzymes and heat shock protein expression.