Liver and spleen transcriptome reveals that Oreochromis aureus under long-term salinity stress may cause excessive energy consumption and immune response.

To investigate the physiological responses of Oreochromis aureus to salinity fluctuations at the molecular level. We used RNA-seq to explore the differentially expressed genes (DEGs) in the liver and spleen of O. aureus at 0, 3, 7 and 11 ppt (parts per thousand) salinity levels. Herein, De novo assembly generated 71,009 O. aureus unigenes, of which 34,607 were successfully mapped to the four major databases. A total of 120 shared DEGs were identified in liver and spleen transcripts, of which 83 were up-regulated and 37 were down-regulated. GO and KEGG analysis found a total of 26 significant pathways, mainly including energy metabolism, immune response, ion transporters and signal transduction. The trend module category of DEGs showed that the genes (e.g., FASN, ODC1, CD22, MRC, TRAV and SLC7 family) involved in the change-stable-change (1) and the constant-change categories (2) were highly sensitive to salinity fluctuations, which were of great value for further study. Based on these results, it would help provide basic data for fish salinity acclimation, and provide new insights into evolutionary response of fish to various aquatic environments in the long-term stress adaptation mechanism.

Expression analysis of immune-associated genes in hemocytes of mud crab Scylla paramamosain under low salinity challenge.

To gain knowledge on the immune response in Scylla paramamosain under low salinity challenge, S. paramamosain we investigated digital gene expression (DEG) in S. paramamosain hemocytes using the deep-sequencing platform Illumina Hiseq XTen. A total of 97,257 high quality unigenes with mean length 786.59 bp were found to be regulated by low salinity challenge, among which 93 unigenes were significantly up regulated, and 71 were significantly down regulated. Functional categorization and pathways analysis of differentially expressed genes revealed that immune signaling pathway including cAMP and cGMP signaling pathway were affected in low salinity stress. Cellular immunity-related genes including low-density lipoprotein receptor-related protein 6 (LRP6) and xanthine dehydrogenase (XDH) were down-regulated, indicating phagocytosis and oxygen dependent mechanism of phagocyte were suppressed in low salinity stress; Humoral immunity-related genes serine proteases and serpins 3 were up- and down-regulated, respectively, suggest that the proPO system was influenced by low salinity significantly; Moreover, processes related to immune response including carbohydrate metabolism, protein synthesis and lipid transport were found differentially regulated, implying the integrity of the immune response in low salinity stress. This study gained comprehensive insights on the immune mechanism of S. paramamosain at low salinity stress at the molecular level. The findings provide a theoretical basis for understanding immune mechanisms of S. paramamosain under low salinity stress, and technical reference for evaluating physiological adaptation in fresh water environment.

Immunocompetence of Penaeus monodon under acute salinity stress and pathogenicity of Vibrio harveyi with respect to ambient salinity.

Salinity is an important environmental factor which usually goes unnoticed in shrimp growout systems. In the present study an attempt was done to analyse the physiological and immunological responses of Penaeus monodon to Vibrio harveyi infection under acute salinity stress. Shrimps were challenged with V. harveyi under 5‰ salinity stress, 35‰ salinity stress and optimal salinity (15‰) conditions by intramuscular injection. A control was maintained without bacterial challenge. Haemolymph was collected from shrimps soon after salinity change before V. harveyi challenge (post salinity change day (PSD) 0), on post challenge day (PCD) 2, 7 and 10. Immune variables viz., total haemocyte count (THC), phenol oxidase activity (PO), Nitroblue tetrazolium salt (NBT) reduction, alkaline phosphatase activity (ALP), acid phosphatase activity (ACP) and metabolic variables viz., total protein (TP), total carbohydrates (TC), total free amino acids (TFAA), total lipids (TL), glucose (Gl) and cholesterol (Ch) were then determined. One way ANOVA followed by Duncan's multiple comparison of the means revealed significant differences between shrimps subjected to different salinity conditions and then challenged with V. harveyi (P < 0.05). Post challenge survival was significantly higher in shrimps maintained at 15‰ and acute salinity stress reduced the survival rate. However, pathogenicity of V. harveyi was slightly higher at 35‰ than at 5‰. At the onset of salinity stress, parameters viz., THC, NBT and ACP significantly reduced and TC increased at 5‰ and 35‰. Following V. harveyi challenge, an overall increase could be observed in metabolic variables on PCD2 and immune variables on PCD2 and 7 in P. monodon maintained at 15‰ compared to the control. Even though an increase in immune response and metabolic variables could be obtained on post challenge days in shrimps under salinity stress compared to the control, it was considerably low in comparison with shrimps maintained at 15‰. Regression analysis proved that NBT, ALP, TP and TL could be proposed as potential stress indicators to evaluate shrimp health status. In brief, it may be concluded from the study that acute salinity changes evoke physiological responses that affect the immunocompetence and metabolic performance of P. monodon against V. harveyi challenge, thereby increasing the susceptibility to infection. Moreover higher salinity enhanced the pathogenicity of V. harveyi.

Gill transcriptomes reveal expression changes of genes related with immune and ion transport under salinity stress in silvery pomfret (Pampus argenteus).

Salinity is a major ecological factor in the marine environment, and extremely important for the survival, development, and growth of fish. In this study, gill transcriptomes were examined by high-throughput sequencing at three different salinities (12 ppt as low salinity, 22 ppt as control salinity, and 32 ppt as high salinity) in an importantly economical fish silvery pomfret. A total of 187 genes were differentially expressed, including 111 up-regulated and 76 down-regulated transcripts in low-salinity treatment group and 107 genes differentially expressed, including 74 up-regulated and 33 down-regulated transcripts in high-salinity treatment group compared with the control group, respectively. Some pathways including NOD-like receptor signaling pathway, cytokine-cytokine receptor interaction, Toll-like receptor pathway, cardiac muscle contraction, and vascular smooth muscle contraction were significantly enriched. qPCR analysis further confirmed that mRNA expression levels of immune (HSP90A, IL-1ß, TNFα, TLR2, IP-10, MIG, CCL19, and IL-11) and ion transport-related genes (WNK2, NPY2R, CFTR, and SLC4A2) significantly changed under salinity stress. Low salinity stress caused more intensive expression changes of immune-related genes than high salinity. These results imply that salinity stress may affect immune function in addition to regulating osmotic pressure in silvery pomfret.

Effect of ammonia stress on immune variables of Pacific white shrimp Penaeus vannamei under varying levels of pH and susceptibility to white spot syndrome virus.

Of late, Pacific white shrimp Penaeus vannamei culture has intensified globally and is a major contributor to the cultured shrimp produced worldwide. Intensification of its culture has led to elevated ammonia concentration during grow-out. Ammonia toxicity is a function of water pH, temperature, salinity and beyond the optimum range, creates stress to cultured aquatic species which can reduce growth, increase susceptibility to diseases and eventually mortality. The present study was aimed at quantifying the toxic effect of total ammonia nitrogen (TAN) (1, 3, 6 & 9 mg/l) and pH levels (6, 8 & 10) individually and in combination on median survival (50% lethal time) of shrimp (8 g) after exposure for 14 days followed by post-stress challenge with white spot syndrome virus (WSSV) for 9 days. Mortality risk factor and the toxicity effect on the immune variables were evaluated. Individual stressors showed a risk factor of 1-13 times, whereas combined treatments considerably increased the risk of dying compared to control. Low survival (15%) was observed in pH6TAN9 and pH10TAN3 treatments and was substantiated by prominent histological obliteration in gills of shrimp. The cumulative mortality in post-stress WSSV challenged trials was 1-5 times and 1-35 times in individual and combination treatments, respectively compared to control. The study revealed that variations in ammonia and pH beyond the optimal range significantly influence the non-specific immune mechanisms in P.vannamei and increases the susceptibility to WSSV especially in combination treatments.