The mammalian target of rapamycin (mTOR) has been shown to be involved in lipopolysaccharide (LPS)-induced immune responses in many mammal cells. Here, we suggest that the mTOR pathway is involved in the intestinal inflammatory responses evoked by LPS treatment in chicken embryos. The intestinal tissue from Specific pathogen free chick embryos was cultured in the presence of LPS for 2 h. Secretory immunoglobulin A (sIgA) concentrations, messenger RNA (mRNA) expression of cytokines, and protein levels of nuclear factor-κB (NF-κB), mitogen-activated protein kinase (MAPK), mTOR and p70 ribosomal S6 kinase (p70S6K) were determined. The results showed that LPS treatment increased sIgA concentrations in a dose-dependent manner. The mRNA levels of interleukine (IL)-6, IL-8, IL-10, tumor necrosis factor-α and Toll-like receptor (TLR) 4 were upregulated by LPS treatment (P<0.05). Lipopolysaccharide increased the phosphorylation of Jun N-terminal kinase (JNK), p38 MAPK and NF-κB (P<0.05) while decreasing the phosphorylation level of mTOR (P<0.05). Supplementation of leucine at doses of 10, 20 and 40 mM dose-dependently decreased sIgA production. Leucine supplementation at 40 mM restored the phosphorylation level of mTOR and p70S6K while suppressing the phosphorylation levels of NF-κB (P<0.05) and partially down-regulating the phosphorylation of p38 MAPK and JNK. The transcription of IL-6 was significantly decreased by leucine supplementation. These results suggested that leucine could alleviate LPS-induced inflammatory responses by down-regulating NF-κB signaling pathway and evoking mTOR/p70S6K signaling pathway, which may involve in the regulation of the intestinal immune system in chicken embryos.

This study was conducted to determine the optimal space allowance for maximizing the growth performance of pigs at each of the following five growth stages (based on BW ranges): stage 1, 11 to 25 kg BW; stage 2, 25 to 45 kg BW; stage 3, 45 to 65 kg BW; stage 4, 65 to 85 kg BW; and stage 5, 85 to 110 kg BW. A total of 1590 crossbred (Landrace×Yorkshire×Duroc) pigs were assigned to one of four treatments at each growth stage, with three replicates each. Pen areas at each growth stage were 6, 11, 16, 19.5 and 20 m2 for stages 1 to 5, respectively. Space allowances for the four treatments at each growth stage were modified by varying the number of pigs per pen (22, 25, 28 and 31 pigs in T1, T2, T3 and T4, respectively). Blood samples were collected on the final day of each growth stage. The average daily gain (ADG) decreased significantly with decreased space allowances at all growth stages, except at stage 2. Average daily feed intake (ADFI) was not significantly affected by space allowances at stages 1 to 4; however, at stage 5, there was a linear effect of space allowance on ADFI. Thus, the feed conversion ratio showed results similar to those for ADG. Serum cortisol concentrations, indicating the level of stress response, increased as space allowances decreased. The highest serum cortisol concentrations were observed in T3 at stages 2 to 5. Serum tumor necrosis factor-α levels were significantly higher in association with a small space allowance than with at large space allowance at stages 2, 4 and 5. Serum interleukin-1β levels also increased in a significant linear manner at every growth stage in pigs reared at a low space allowance, except at stage 4 (P=0.068). This study found that limited space allowance decreases the growth performance of pigs and induces stress and inflammatory responses. We confirmed that no significant effect of space allowance on growth performance and serum cortisol concentrations are observed between T1 and T2 across all growth stages. We suggest that the optimal space allowances for pigs according to their BW are as follows: 0.24, 0.44, 0.64, 0.78 and 0.80 m2/pig for BWs of 11 to 25, 25 to 45, 45 to 65, 65 to 85 and 85 to 115 kg, respectively.

Global warming is a challenge to animal health, because of increased heat stress, with subsequent induction of immunosuppression and increased susceptibility to disease. Toll-like receptors (TLR) are pattern recognition receptors that act as sentinels of pathogen invasion and tissue damage. Ligation of TLRs results in a signaling cascade and production of inflammatory cytokines, which eradicate pathogens and maintain the health of the host. We hypothesized that the TLR signaling pathway plays a role in immunosuppression in heat-stressed pigs. We explored the changes in the expression of TLR2, TLR4 and the concentration of acute inflammatory cytokines, such as IL-2, IL-8, IL-12 and IFN-γ in Bama miniature pigs subjected to 21 consecutive days of heat stress, both in vitro and in vivo models. The results showed that heat stress induced the upregulation of cortisol in the plasma of pigs (P<0.05); TLR4 mRNA was elevated, but IL-2 was reduced in peripheral blood mononuclear cells (PBMC, P<0.05). The white blood cell count and the percentage of granulocytes (eosinophilic+basophilic) decreased significantly in heat-stressed pigs (P<0.05). In the in vitro model (PBMC heat shocked for 1 h followed by a 9 h recovery period), TLR2 and TLR4 mRNA expression also increased, as did the concentration of IL-12 in supernatants. However, IFN-γ was significantly reduced in PBMC culture supernatants (P<0.05). We concluded that a consecutive heat stress period elevated the expression of TLR2 and TLR4 in PBMC and increased the plasma levels of inflammatory cytokines. These data indicate that TLR activation and dysregulation of cytokine expression in response to prolonged heat stress may be associated with immunosuppression and increased susceptibility to antigenic challenge in Bama miniature pigs.