This experiment aimed to study the effects on liver tissue structure and immune performance of grass carp Ctenopharyngodon idellus when the common reed Phragmites australis is in its diet, instead of wheat Triticum aestivum L. Seventy-two healthy grass carps (145.52 ± 2.56 g) were randomly divided into three groups according to their body weight. Fishes in each group were fed an essential diet with 0% (control group), 50% and 100% (test group) common reed, instead of wheat, respectively. After feeding for 41 days, the changes of serum biochemical indices, liver tissue structure and immune related indices of grass carp were detected. The results showed that, compared with the control group, the activities of serum alanine transaminase (ALT) and aspartate transaminase (AST) in the substitution groups were significantly increased (p < 0.05), but still at a normal level. The contents of total protein, albumin and globulin did not change significantly (p > 0.05). Compared with the control group, the liver cells of grass carp in the substitution groups had clear boundaries, tight arrangement and less vacuolation. The contents of serum interleukin-1 (IL-1) and complement 3 (C3) in the 100% substitution group were significantly higher than those in the control group (p < 0.05), and the contents of liver IL-1 and total complement (CH50) in the 100% substitution group were significantly higher than those in the control group (p < 0.05). The contents of IL-1, C3 in serum and IL-1, CH50 in liver in the 50% substitution group were significantly higher than those in the control group (p < 0.05). The mRNA relative expression levels of C3, IL-1, MHC-I and interferon (IFN) in the head-kidney, kidney, liver and spleen of grass carp were significantly affected by feeding the grass carp with different common reed substitution ratios (p < 0.05). In summary, common reed, instead of wheat, in feed can improve the liver tissue structure, and increase the non-specific immune response level, of grass carp.
Interferon regulatory factors (IRFs) have been confirmed as vital transcription factors for the immune system, which play a certain role in anti-infection defense, immune regulation, hematopoietic cell development, and maturation of the immune system. In this study, the cDNA of Interferon regulatory factor 5 (IRF5) was cloned from black-spotted frog, Pelophylax nigromaculatus, and termed PnIRF5. The results indicated, the full-length cDNA of PnIRF5 was 2090 bp, and a putative protein of 504 amino acids was encoded. The results of protein domain prediction suggested that PnIRF5 exhibited a DNA-binding domain (DBD), a middle region (MR), an IRF association domain (IAD), a virus activated domain (VAD), as well as two nuclear localization signals (NLSs). Phylogenetic analysis clustered PnIRF5 into the amphibian IRF5 subgroup in vertebrate IRF5 group. The results of qPCR show that PnIRF5 mRNA was expressed in all examined tissues, with higher levels were identified in the kidney, intestine, and lung. The gene expression of PnIRF5 was analyzed in kidney, spleen, and liver of black-spotted frog after challenged with Elizabethkingia miricola, and the maximum expression level of PnIRF5 in liver, spleen, and kidney were 1.80, 2.33 and 2.88 times that at 0 days post challenged, respectively. As indicated by the results, the PnIRF5 protein may take on critical significance in protecting black-spotted frog from E. miricola infection, and they may play a certain role in the immune response of black-spotted frog against bacteria.
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