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Intermittent mild cold stimulation improves the immunity and cold resistance of spleens in broilers

ABSTRACT: In order to investigate the effect of intermittent mild cold stimulation (IMCS) on immune function of spleens and adaptability to cold stress in broilers, 400 healthy 1-day-old Ross-308 chickens were divided into 5 groups: CC (control) reared in normal thermal environment from 1 to 49 d; CS3, CS4, CS5, and CS6 (treatments) raised at 3°C below the temperature of CC for 3, 4, 5, or 6 h at 1-d intervals from 15 to 35 d, respectively. Subsequently, CS3-6 was raised at 20°C from 36 to 49 d. At 50 d, all groups were exposed to acute cold stress (ACS) for 12 h. The spleen immunity index at 22, 29, 36, 43, and 49 d, expression levels of toll-like receptors (TLRs), cytokines and immunoglobulins at 22, 43, and 49 d and heat shock proteins (HSPs) before and after ACS at 50 d were examined. The spleen index of broilers aged 22 to 49 d did not differ between CS and CC (P > 0.05), and the spleen index of CS5 was higher than that of CS3 at 49 d (P < 0.05). The mRNA levels of TLR5, TLR15, TLR21, and IL-2 in CS3, TLR3, TLR4, TLR15, TLR21, IL-2, IL-6, and IFN-ϒ in CS4, TLR1, TLR3, TLR4, TLR21, IL-2, IFN-a, IFN-ϒ, IgA, and IgG in CS6, but all TLRs, immunoglobulins and cytokines except IFN-ϒ in CS5 differential expressed stably compared with CC at 43 and 49 d (P < 0.05). Compared with Pre-ACS, the mRNA levels of HSP60, HSP70, and HSP90 were upregulated in CS after ACS (P < 0.05). Except for HSP90 mRNA and HSP70 protein in CS6, and HSP90 protein in CS3, the levels of HSPs after ACS in all treatment groups were higher than those in CC (P < 0.05), and the highest HSPs levels after ACS were found in CS5. We concluded that IMCS could enhance immunity of spleens and adaptability to ACS in broilers, besides CS5 was the optimal program.

SUBMITTER: Xue G

PROVIDER: S-EPMC8554259 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:A total of 240 healthy 1-day-old Ross 308 male broilers were randomly divided into 3 groups (CS0 group, CS3 group, and CS6 group), with 5 replicates in each group and 16 broilers in each replicate, in order to evaluate the effects of intermittent mild cold stimulation (IMCS) on the intestinal immune function and anti-cold stress ability of broilers after acute cold stress. The mRNA expression levels of cytokines and Toll-like receptors (TLRs) in the duodenum and jejunum were detected at the end of cold stimulation (36 d), 2 wk after recovery (50 d), and after acute cold stress (Y6). In addition, the mRNA and protein expression levels of heat shock proteins (HSPs) were measured before and after acute cold stress. The experimental data were statistically processed using 1-way ANOVA and Duncan's multiple comparisons. The results showed that the mRNA expression levels of IL2, IL8, IFN γ, TLR7, and TLR21 in the duodenum and IL2 and IFN γ in jejunum were significantly higher in the CS6 group than in the CS0 and CS3 groups at 36 d (P < 0.05). All TLR levels in the jejunum were significantly lower in the CS3 group than in the CS0 and CS6 groups at 36 d (P < 0.05). After 6 h of acute cold stress, in the duodenum, the mRNA expression levels of IL6 and IL8 were significantly decreased in the CS0 and CS6 groups compared to levels at 50 d (P < 0.05), while levels in the CS3 group remained stable (P > 0.05). Compared with 50 d, the expression level of HSP mRNA in the jejunum in the CS3 group was relatively stable compared to that in the CS0 and CS6 groups after acute cold stress (P > 0.05). At the protein level, the HSP60 expression level in the duodenum and HSP40, HSP60, and HSP70 expression levels in the jejunum were significantly higher in the CS3 group than in the CS0 and CS6 groups after acute cold stress (P < 0.05). In conclusion, cold stimulation training at 3℃/3 h lower than the conventional feeding temperature can improve the intestinal immune function and anti-stress ability of broilers.

Project description:Improving immune function is an important indicator for establishing cold adaptation in broilers. In the study, to explore the effects and molecular mechanisms of intermittent and mild cold stimulation (IMCS) on the immune function of broilers, CIRP and TRPM8, induced by cold stimulation, as well as the NF-κB and MAPK pathways which play an important role in immune response, were selected to investigate. A total of 192 one-day-old broilers (Ross 308) were selected and randomly divided into the control group (CC) and the cold stimulation group (CS). The broilers in CC were raised at normal feeding temperature from d 1 to 43, while the broilers in CS were subjected to cold stimulation from day 15 to 35, with a temperature 3 °C below that of the CC group for 5 h, at 1 d intervals. The results showed that IMCS had little effect on the broiler hearts, and the myocardial structure was not damaged. On d 22, IMCS significantly increased the mRNA levels of CIRP, TRPM8, P65, P38, COX-2, TNF-α, IFN- γ, IL-6, IL-10, and the protein levels of CIRP, P65, P38, IL-1β and iNOS in the hearts, and the levels of CIRP and all cytokines in the serum (P ≤ 0.05). The mRNA and protein levels of IκB-α were significantly reduced (P ≤ 0.05). On d 36, the mRNA levels of TRPM8, P65, ERK, and IL-10 in the hearts and the content of COX-2 in the serum in CS were increased significantly (P ≤ 0.05), while the mRNA levels of IκB-α, P38, and IL-1β were decreased significantly (P ≤ 0.05). On d 43, IMCS significantly upregulated the mRNA levels of TRPM8, IFN- γ, IL-4, IL-6, IL-10, and the protein levels of IκB-α, P38, and the levels of iNOS, TNF-α, IL6 and IL10 in the serum (P ≤ 0.05); whereas it significantly downregulated CIRP, JNK, P38, iNOS, TNF-α mRNA levels, and CIRP, P65, ERK, JNK, IL1β and iNOS protein levels (P ≤ 0.05). Therefore, IMCS can enhance broiler immune function through co-regulation of CIRP and TRPM8 on the NF-κB and MAPK pathways, which facilitate the cold adaptation in broilers.

Project description:To investigate the potential protective effect of prior cold stimulation on broiler intestine induced by acute cold stress (ACS). A total of 384 one-day-old broilers were divided into control (CON), ACS, cold stimulation Ⅰ (CS3+ACS), and cold stimulation Ⅱ (CS9+ACS) groups. Broilers in CON and ACS groups were reared normally, and birds in CS3+ACS and CS9+ACS groups were reared at 3℃ and 9℃ below CON group for 5 h, respectively, on alternate days from d 15 to 35. Broilers in ACS, CS3+ACS, and CS9+ACS groups were subjected to 10℃ for 24 h on d 43. Eventually, small intestine tissues were collected for histopathological observation and indexes detection. The results showed that intestinal tissues in all ACS-broilers exhibited inflammatory cell infiltrates, microvilli disruption, reduced villus length in jejunum and increased crypt depth in jejunum and ileum. Whereas these phenomena were relatively light in CS3+ACS group. Compared to CON group, mRNA expression of the TLR4/MyD88/NF-κB pathway-related genes (TLR4, MyD88, NF-κBp65, COX-2, iNOS, PTGEs, TNF-α), Th1/Th17-derived cytokines (IL-1β, IL-2, IL-8, IL-12, IFN-γ, IL-17), and HSPs (HSP40, HSP60, HSP70, HSP90) was upregulated (P < 0.05), and that of Th2-deviated cytokines (IL-4, IL-6, IL-10, IL-13) and IκBα was downregulated (P < 0.05) in small intestine in almost all ACS-broilers. Compared to ACS group, mRNA expression of most of the TLR4/MyD88/NF-κB pathway-related genes, Th1/Th17-derived cytokines, and HSPs was downregulated and that of Th2-derived cytokines was upregulated in CS3+ACS group (P < 0.05). Protein expression levels of TLR4, MyD88, p-p65/p65, p-IκBα/IκBα, IKK, TNF-α, IL-1β, IL-10, and HSPs were similar to their mRNA expression. The concentration of sIgA and activities of CAT, SOD, and GSH-px were decreased and MDA and H2O2 were increased in ACS and CS9+ACS groups compared to CON group (P < 0.05). Therefore, cold stress caused oxidative stress and inflammation, leading to gut immune dysfunction; while mild cold stimulation at 3℃ below normal rearing temperature alleviated cold stress-induced intestinal injure and dysfunction by modulating the TLR4/MyD88/NF-κB pathway in broilers.

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Intermittent mild cold stimulation improves the immunity and cold resistance of spleens in broilers

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