ALBERT

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Ying-Yu Tang, Qiu-Ning Liu, Cheng Wang, Ting-Ting Yang, Bo-Ping Tang, Chun-Lin Zhou, Li-Shang Dai〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉〈em〉Procambarus clarkii〈/em〉 is one of the most important aquatic invertebrates in China and has high commercial value. However, aquaculture has suffered great economic loss due to outbreaks of infectious diseases in 〈em〉P〈/em〉. 〈em〉clarkii〈/em〉. To identify red swamp crayfish related proteins involved in the response to bacterial infection, we analysed immune-related proteins following lipopolysaccharide (LPS) stimulation by quantitative proteomics. The proteome of the hepatopancreas of 〈em〉P〈/em〉. 〈em〉clarkii〈/em〉 challenged with LPS and phosphate-buffered saline was analysed to evaluate the immune response. Based on liquid chromatography coupled with tandem mass spectrometry, 16 upregulated and 29 downregulated proteins were identified. A Gene Ontology analysis demonstrated 5 biological process, 11 cellular component, and 6 molecular function subcategories. The Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis indicated that the identified proteins were mainly involved in metabolism, phagosome, and ribosome. Real-time quantitative reverse transcription-PCR revealed that eight immune-related genes were upregulated after LPS stimulation compared to the control. Taken together, the data enhance our understanding of the immune response of crayfish to LPS.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Mahmoud A.O. Dawood, Safaa E. Abdo, Mahmoud S. Gewaily, Eman M. Moustafa, Moustafa S. SaadAllah, Marwa F. AbdEl-kader, Awatef H. Hamouda, Amira A. Omar, Rasha A. Alwakeel〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The protective role of β-glucan (BG) on liver function, histopathology, immune and antioxidant related gene expressions in Nile tilapia exposed to subacute deltamethrin (DLM) was investigated for 30 days. Fish (28.18 ± 1.34 g) of the 1st and 2nd groups fed the control diet, while the 3rd and 4th groups fed BG at 0.5 g/kg and the 2nd and 4th groups were exposed to DLM (15 μg/L) in rearing water. DLM-treated fish displayed a considerable increase in blood biochemical parameters (creatinine, urea and bilirubin) as well as hepatic enzymes (ALP, AST and ALT) (〈em〉P〈/em〉 〈 0.05). Blood total protein, globulin, albumin, WBCs, RBCs, Hb, phagocytic index, phagocytic and lysozyme activities were significantly decreased in fish subjected to DLM (〈em〉P〈/em〉 〈 0.05). Fish fed BG showed significantly the lowest cortisol and glucose levels, while fish exposed to DLM without feeding BG showed the highest cortisol and glucose levels (〈em〉P〈/em〉 〈 0.05) after 15 and 30 days. Additionally, DLM toxicity caused downregulation in antioxidant (〈em〉CAT〈/em〉 and 〈em〉GPx〈/em〉) and immune (〈em〉IL-1β〈/em〉 and 〈em〉IL-8〈/em〉) related gene expressions, while and 〈em〉IFN-γ〈/em〉, 〈em〉HSP70〈/em〉 and 〈em〉CASP3〈/em〉 were upregulated. The histopathological examination of Nile tilapia exposed to DLM revealed damage in gills, intestine, spleen and liver which confirmed the toxic effects. Conversely, BG presented protective effects and restored the above-mentioned parameters when fish exposed to DLM and fed BG. Thus, BG supplementation exhibited defensive effects against DLM toxicity in Nile tilapia through improving blood biochemical responses, immune, and antioxidant related gene expressions as well as histopathological effects.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Junjie Nie, Zhixue Yu, Defu Yao, Fan Wang, Chunhua Zhu, Kaihui Sun, Jude Juventus Aweya, Yueling Zhang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The histone deacetylase, sirtuin 6 (SIRT6), plays an essential role in the regulation of oxidative stress, mitochondrial function and inflammation in mammals. However, the specific role of SIRT6 in invertebrate immunity has not been reported. Here, we characterized for the first time, a sirtuin 6 homolog in 〈em〉Litopenaeus vannamei〈/em〉 (〈em〉Lv〈/em〉SIRT6), with full-length cDNA of 2919 bp and 1536 bp open reading frame (ORF) encoding a putative protein of 511 amino acids, which contains a typical SIR2 domain. Sequence and phylogenetic analysis revealed that 〈em〉Lv〈/em〉SIRT6 shares a close evolutionary relationship with SIRT6 from invertebrates. Real-time quantitative PCR analysis of 〈em〉Lv〈/em〉SIRT6 transcripts revealed that they were ubiquitously expressed in shrimp and induced in hepatopancreas and hemocytes upon challenge with 〈em〉Vibrio parahaemolyticus〈/em〉, 〈em〉Streptococcus iniae〈/em〉, lipopolysaccharide (LPS), and white spot syndrome virus (WSSV), suggesting the involvement of 〈em〉Lv〈/em〉SIRT6 in shrimp immune response. Moreover, knockdown of 〈em〉Lv〈/em〉SIRT6 decreased mitochondrial membrane potential and increased total ROS level in hemocytes, especially upon 〈em〉V. parahaemolyticus〈/em〉 challenge. Depletion of 〈em〉Lv〈/em〉SIRT6 also increased hemocytes apoptosis in terms of decreased expression of pro-survival 〈em〉Lv〈/em〉Bcl-2, but increased expression of pro-apoptotic 〈em〉Lv〈/em〉Bax and 〈em〉Lv〈/em〉Cytochrome C, coupled with high 〈em〉Lv〈/em〉Caspase3/7 activity. Shrimp were rendered more susceptible to 〈em〉V. parahaemolyticus〈/em〉 infection upon 〈em〉Lv〈/em〉SIRT6 knockdown. Taken together, our present data suggest that 〈em〉Lv〈/em〉SIRT6 plays an important role in shrimp immune response by modulating hemocytes ROS production and apoptosis during pathogen challenge.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Lang Lang, Minnan Bao, Weixin Jing, Wei Chen, Lan Wang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉 〈p〉Toll is essential in innate immune system which is important for defense against bacterial, fungal and viral infections in invertebrates. Our previous study showed that cadmium (Cd) could change the expression pattern of ShToll3 in the epithelium (gills and midgut from the freshwater crab 〈em〉Sinopotamon henanense〈/em〉) infected by 〈em〉Aeromonas hydrophila〈/em〉. To investigate the diverse innate immune roles of crustacean homolog Tolls, in this study, we cloned 〈em〉Shtoll1〈/em〉 from 〈em〉S. henanense〈/em〉. The full-length cDNA of 〈em〉Shtoll1〈/em〉 was 4746 bp, with an ORF of 3033 bp encoding a putative protein of 111 amino acids, a 5′-untranslated region of 255 bp and a 3′-untranslated region of 1713 bp. Phylogenetic analysis showed that ShToll1 was clustered into the group of DmToll1, DmToll 4 and DmToll 5. In addition, the tissue distribution results showed that 〈em〉Shtoll1〈/em〉 was expressed widely in different tissues, with the highest expression in heamocytes. Besides, 〈em〉Shtoll1〈/em〉 expressions were upregulated in heamocytes and hepatopancreas after 〈em〉A. hydrophila〈/em〉 infection. At the same time, the increase of 〈em〉Shtoll1〈/em〉 expressions were examined in heamocytes in response to Cd exposure and 〈em〉A. hydrophila〈/em〉 infection in combination. Through western blotting and immunohistochemical analysis, the ShToll1 expressions in heamocytes were increased in response to 〈em〉A. hydrophila〈/em〉 and Cd independently as well as in combination. Moreover, the mRNA level of three antimicrobial peptides (AMPs) 〈em〉alf5, alf6,〈/em〉 and 〈em〉c-lys〈/em〉, which possibly responded to Cd and 〈em〉A. hydrophila〈/em〉 stimulation through 〈em〉Shtoll1,〈/em〉 were analyzed.〈/p〉 〈p〉Thus, we conclude that Cd expand the susceptibility of ShToll1 to 〈em〉A. hydrophila〈/em〉 infection in heamocytes. This suggest that ShToll1 may contribute to the innate immune defense of 〈em〉S. henanense〈/em〉 against 〈em〉A. hydrophila〈/em〉 and Cd in heamocytes.〈/p〉 〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 13 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology〈/p〉 〈p〉Author(s): A. Panigrahi, R.R. Das, M.R. Sivakumar, A. Saravanan, C. Saranya, N.S. Sudheer, K.P. Kumaraguru Vasagam, P. Mahalakshmi, S. Kannappan, G. Gopikrishna〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Effect of bio-augmentation of 〈em〉Bacillus spp〈/em〉 in biofloc on growth, survival and immunity in Indian white shrimp 〈em〉Penaeus indicus〈/em〉 was evaluated. Nine 〈em〉Bacillus〈/em〉 strains were isolated and screened individually as well as in the form of a consortia. To maintain a C:N ratio of 12:1 a blend of carbohydrate sources was used. Bio-augmentation with bacterial consortium and 〈em〉Virgibacillus sp.〈/em〉 produced improved growth and immunity. Shrimp survival ranged from 80 to 95% among treatments. Production was higher (35%) in the biofloc tanks with an average body weight (ABW) of 10.89 ± 1.2 g. On evaluating the immune responses, it was found that trypsin significantly (P 〈 0.05) enhanced Prophenoloxidase (PO) activity in 〈em〉Lysinibacillus〈/em〉, 〈em〉Bacillus cereus〈/em〉, 〈em〉Bacillus licheniformis〈/em〉 and 〈em〉Bacillus subtilis〈/em〉 bio-augmented groups. Laminarin induced PO activity was observed in groups supplemented with 〈em〉Oceanobacillus sp.〈/em〉, 〈em〉Bacillus sp.〈/em〉and 〈em〉Bacillus megaterium〈/em〉. The lysozyme (LZ) activity was significantly (P 〈 0.05) higher in 〈em〉B. cereus〈/em〉 and Microbial Consortia (MC), while other treatments were less effective. Total hemocyte count (THC) significantly (P 〈 0.05) increased in all treatment groups compared to the control. Hyaline hemocyte (HH) count was significantly (P 〈 0.05) higher in the control group (14.43%). Semi granular hemocytes (SGH) was higher in groups treated with 〈em〉Lysinibacillus, Bacillus sp., B. licheniformis〈/em〉 and 〈em〉B. subtilis.〈/em〉 The granular hemocyte (GH) count was significantly (P 〈 0.05) higher in 〈em〉Virgibacillus sp.〈/em〉, 〈em〉B. cereus〈/em〉, 〈em〉B.megaterium〈/em〉 and 〈em〉Oceanobacillus sp.〈/em〉 The biofloc alone (BF), treated and augmented with 〈em〉B. megaterium〈/em〉 significantly (P 〈 0.05) increased phagocytic activity. Highly significant phagocytic index (PI) was observed in bio-augmented groups, BF and MC. The relative expression levels of immune genes were found to be significantly up-regulated in shrimps grown in bio-augmented groups. Enhanced immunological parameters implies that bio-augmentation of biofloc with 〈em〉Bacillus spp.〈/em〉 improved immunity in shrimps. Hence, bio-augmentation of probiotics in biofloc may be useful in improving culture conditions to produce 〈em〉P. indicus.〈/em〉〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Jessica L. Piesz, Sarah E. Barker, Ian R. Bricknell〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The ectoparasite, 〈em〉Lepeophtheirus salmonis〈/em〉 (Kroyer 1837)〈em〉,〈/em〉 is effective at avoiding elimination from its host, Atlantic salmon, 〈em〉Salmo salar〈/em〉 L., by inhibiting the recruitment of immune cells to the site of attachment. In other ectoparasitic arthropods, numerous factors have been identified that bind or neutralize chemokines preventing their interaction with receptors on the surfaces of immune cells. To determine if 〈em〉L. salmonis〈/em〉 is utilizing a similar mechanism of immune modulation, the chemotactic activity of peripheral blood leukocytes (PBL) to leukotriene B4 (LTB4) and the secreted/excreted products (SEPs) of the sea louse were investigated 〈em〉in vitro〈/em〉. The results showed that incubation of LTB4 with SEPs reduced leukocyte migration compared to LTB4 immune stimulation alone. Data suggests that one of the mechanisms 〈em〉L. salmonis〈/em〉 may be using to regulate immune cell recruitment in Atlantic salmon is by inhibiting or neutralizing the activity of chemokines.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Li-Jie Huo, Ming-Chong Yang, Jin-Xing Wang, Xiu-Zhen Shi〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉ATPase Inhibitory Factor 1 (IF1) is a mitochondrial protein that functions as a physiological inhibitor of F〈sub〉1〈/sub〉F〈sub〉0〈/sub〉-ATP synthase. In the present study, a mitochondrial ATPase inhibitor factor 1 (〈em〉Mj〈/em〉ATPIF1) was identified from kuruma shrimp (〈em〉Marsupenaeus japonicus〈/em〉), which was demonstrated to participate in the viral immune reaction of white spot syndrome virus (WSSV). 〈em〉Mj〈/em〉ATPIF1 contained a mitochondrial ATPase inhibitor (IATP) domain, and was widely distributed in hemocytes, heart, hepatopancreas, gills, stomach, and intestine of shrimp. 〈em〉MjATPIF1〈/em〉 transcription was upregulated in hemocytes and intestines by WSSV. WSSV replication decreased after 〈em〉MjATPIF1〈/em〉 knockdown by RNA interference and increased following recombinant 〈em〉Mj〈/em〉ATPIF1 protein injection. Further study found that 〈em〉Mj〈/em〉ATPIF1 promoted the production of superoxide and activated the transcription factor nuclear factor kappa B (NF-κB, Dorsal) to induce the transcription of WSSV RNAs. These results demonstrate that 〈em〉Mj〈/em〉ATPIF1 benefits WSSV replication in kuruma shrimp by inducing superoxide production and NF-κB activation.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Muting Yan, Zhihao Liu, Kaihang Xu, Wenjing Wang, Lanfen Fan, Han Gong〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Previous studies have indicated that white spot syndrome virus (WSSV) infection induces apoptosis in many shrimp organs. However, the mechanism by which WSSV causes host apoptosis remains largely unknown. In this study, we demonstrated the function of wsv152, the first mitochondrial protein identified as encoded by WSSV. Glutathione S-transferase pulldown and co-immunoprecipitation analysis revealed that wsv152 interacts with the shrimp mitochondrial protein cytochrome 〈em〉c〈/em〉 oxidase 5a (COX5a), a subunit of the COX complex. We also found that wsv152 expression significantly increased the rate of apoptosis, suggesting a role of wsv152 in WSSV-induced apoptosis in shrimp. Knockdown of wsv152 〈em〉in vivo〈/em〉 led to downregulation of several apoptosis-related shrimp genes, including cytochrome 〈em〉c〈/em〉, apoptosis-inducing factor and caspase-3. Suppression of wsv152 also resulted in significant reductions in the number of WSSV genome copies in tissues and in the mortality of WSSV-infected shrimp. Together, these results suggest that wsv152 targets host COX5a and is associated with the expression profiles of apoptosis-related shrimp genes. Wsv152 is likely also involved in WSSV-induced apoptosis, thereby facilitating virus infection and playing a complex role in WSSV pathogenesis.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Sen Xu, Ming Jing, Wen-Ying Liu, He Dong, De-Min Kong, Ya-Ru Wang, Han-Han Zhang, Zhen Yue, You-Jie Li, Fei Jiao, Shu-Yang Xie〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉L-type lectins (LTLs) belong to the lectin family and are characterized by a conserved structural motif in their carbohydrate recognition domain. LTLs are homologous to leguminous lectins. In this study, we identified and functionally characterized an LTL from kuruma shrimp 〈em〉Marsupenaeus japonicus〈/em〉. We designated this LTL as 〈em〉Mj〈/em〉LTL2. 〈em〉Mj〈/em〉LTL2 contains a signal peptide, a Lectin_leg domain, a coiled coil, and transmembrane domain. 〈em〉Mj〈/em〉LTL2 is distributed in hemocytes, heart, hepatopancreas, gill, stomach, and intestine; higher expression levels are seen in hemocytes and the hepatopancreas than in other tissues. 〈em〉Mj〈/em〉LTL2 was upregulated following challenge of shrimp with 〈em〉Vibrio anguillarum〈/em〉 and white spot syndrome virus (WSSV). 〈em〉Mj〈/em〉LTL2 can agglutinate several bacteria without Ca〈sup〉2+〈/sup〉. In addition, 〈em〉Mj〈/em〉LTL2 could bind to several Gram-positive and -negative bacteria by binding to their lipopolysaccharide and peptidoglycan. However, 〈em〉Mj〈/em〉LTL2 could not enhance the clearance of 〈em〉V. anguillarum〈/em〉 in vivo. In the presence of WSSV infection, 〈em〉Mj〈/em〉LTL2 knockdown by RNA interference resulted in a 7-day lower cumulative mortality of 〈em〉M. japonicus〈/em〉. Moreover, less 〈em〉VP19, VP24, VP26,〈/em〉 and 〈em〉VP2〈/em〉8 mRNAs were extracted from the hemocytes of 〈em〉Mj〈/em〉LTL2 knockdown shrimp than from the control. These results suggest that 〈em〉Mj〈/em〉LTL2 is involved in immune responses in shrimp.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Yiyi Yan, Xingchen Huo, Taoshan Ai, Jianguo Su〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉As one of the most important fish in freshwater aquaculture, gibel carp (〈em〉Carassius auratus gibelio〈/em〉) is easily susceptible to Cyprinid herpesvirus 2 (CyHV-2). Immersion vaccination has attracted many researchers due to its simple operation in preventing infectious diseases. However, the unavoidable disadvantage is that the immersion vaccine must be used with adjuvants to get a better performance. In this study, gibel carps were vaccinated by a 60 min bath in a β-propiolactone-inactivated Cyprinid herpesvirus 2, mixed with DTT, β-glucan, anisodamine and scopolamine, respectively. After immunization, the fishs were challenged by CyHV-2 in 2 weeks. By analyzing pathological section, we found that β-glucan, anisodamine and scopolamine groups protected the gibel carp compared to the control group, which was consistent with the trend of survival rate. Specifically, β-glucan group in serum appeared best on lysozyme, TSOD and complement C3. Real time quantitative RT-PCR results demonstrated that in both spleen and head kidney tissues, mRNA expressions of typical Th1 immune response cytokines IL-2 and IFN-γ2 in β-glucan group and anisodamine group were significantly higher than other groups and the level of immunoglobulins related to systemic immunity (IgM) and mucosal immunity (IgZ) were also enhanced in the immune period. DTT group slightly affected immune gene and serum enzyme activity, while did not show an adjuvant effect on survival rate. In addition, four adjuvant groups could obviously inhibit CyHV-2 replication. This study explored and proved the good efficiency of β-glucan or anisodamine as immersion immune adjuvant and also provided reference for improving the efficiency of immersion immunity.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Jing Wang, Wenjing Hong, Fei Zhu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Astakine is a crucial factor in the proliferation and differentiation of hematopoietic stem cells and is directly involved in hematopoiesis in crustaceans. To assess the role of Astakine in the innate immune system of 〈em〉Scylla paramamosain〈/em〉, the immune responses in healthy and Astakine-inhibited 〈em〉S. paramamosain〈/em〉 were investigated in the present study. The RNA transcripts of 〈em〉Astakine〈/em〉 were widely distributed in all examined tissues, with significantly higher levels of expression in hemocytes of both healthy and challenged 〈em〉S. paramamosain〈/em〉 with 〈em〉Vibrio alginolyticus〈/em〉 and WSSV. When 〈em〉Astakine〈/em〉 was knocked down by RNA interference technology, immune-related genes, including 〈em〉Janus kinase〈/em〉, 〈em〉prophenoloxidase〈/em〉, 〈em〉hemocyanin〈/em〉, 〈em〉β-actin〈/em〉, 〈em〉myosin II essential light chain-like protein〈/em〉, 〈em〉signal transducer and activator of transcription〈/em〉, 〈em〉Relish〈/em〉, and 〈em〉C-type-lectin〈/em〉, were significantly down-regulated in hemocytes. The levels of phenoloxidaseactivity (PO), total hemocyte counts (THC) and hemocyte proliferation decreased significantly in hemocytes of Astakine-dsRNA treated 〈em〉S. paramamosain〈/em〉. After being challenged with 〈em〉V. alginolyticus〈/em〉 and WSSV, the THC decreased significantly and the levels of hemocyte apoptosis increased significantly in Astakine-dsRNA treated 〈em〉S. paramamosain〈/em〉 in comparison with those in infected groups without Astakine-dsRNA treatment. After being challenged with WSSV, the WSSV copies were significantly lower in Astakine-dsRNA treated groups than those in the WSSV infection group, which suggested that knockdown of 〈em〉Astakine〈/em〉 was not conductive to WSSV replication and this might be associated with the decreasing THC. The results of survival analysis showed that the survival rate of 〈em〉V. alginolyticus〈/em〉 or WSSV infected 〈em〉S. paramamosain〈/em〉 decreased significantly following 〈em〉Astakine〈/em〉 knockdown. These results suggested that RNA interference of 〈em〉Astakine〈/em〉 might weaken the resistance of 〈em〉S. paramamosain〈/em〉 to 〈em〉V. alginolyticus〈/em〉 or WSSV infection. The weaken resistivity after knockdown 〈em〉Astakine〈/em〉 might be related to the changes of important immune-related gene expression, THC, PO activity, proliferation and apoptosis of hemocytes.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Yunkun Li, Jiabei Han, Jiayu Wu, Dong Li, Xixi Yang, Anqi Huang, Guixian Bu, Fengyan Meng, Fanli Kong, Xiaohan Cao, Xingfa Han, Xiaofu Pan, Shiyong Yang, Xianyin Zeng, Xiaogang Du〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Quantification real-time PCR (qRT-PCR) is a common method in analysis of gene expression, but the stable reference genes for the normalization analysis have not been appreciated before identifying expression pattern of genes in teleost fishes. In this study, we selected eight candidate reference genes (〈em〉18S〈/em〉, 〈em〉Actin〈/em〉, 〈em〉EF-1α〈/em〉, 〈em〉40S〈/em〉, 〈em〉B2M〈/em〉, 〈em〉TUBA〈/em〉, 〈em〉UBCE〈/em〉 and 〈em〉GAPDH〈/em〉) basing on transcriptome analysis and the traditional housekeeping genes, and analyzed the stability of the reference genes in spleen, head kidney and head kidney leukocytes (HKL) after pathogen challenge in 〈em〉Schizothorax prenanti〈/em〉 (〈em〉S. prenanti〈/em〉). Three common programs (geNorm, NormFinder and Bestkeeper) were used to evaluate the stability of the candidate reference genes. Two reference genes, 〈em〉Actin〈/em〉 and 〈em〉EF-1α〈/em〉 presented higher stability, while 〈em〉18S〈/em〉 and 〈em〉GAPDH〈/em〉 were the lower stable genes, both in 〈em〉in vitro〈/em〉 and 〈em〉in vivo〈/em〉. An important immune gene, 〈em〉toll-like receptor 22a〈/em〉 (〈em〉TLR22a〈/em〉), was selected to validate the stability of the proposed reference genes (〈em〉Actin〈/em〉 and 〈em〉EF-1α〈/em〉) across different experiment treatments. The results reveal that 〈em〉Actin〈/em〉 and 〈em〉EF-1α〈/em〉 are quite suitable reference genes for the normalization analysis. Otherwise, using the most stable gene 〈em〉Actin〈/em〉 to validate the reliable of transcriptome data showed the high correlation between the fold change of transcriptome data and qRT-PCR data. In conclusion, our study not only acquired the suitable reference gene for the qRT-PCR assay under specific experiment condition, but also provided a comprehensive method to evaluate and validate the reference gene based on transcriptome analysis in teleost fishes.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Xiang Sun, Peng Jin, Qin Liu, Qiyao Wang, Yuanxing Zhang, Xiaohong Liu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Inactivated vaccines are often applied with adjuvants in commercial fish farming. Although some mineral or non-mineral oil adjuvants show efficient improvement with inactivated vaccines, but sometimes bring side effects such as tissue adhesion and granulomatous lesion at the injection site. CpG ODN is a novel type of soluble adjuvant which has been proved to possess excellent advantages in fish vaccine development. In this study, we designed a tandem sequence of CpG ODN synthesized in plasmid pcDNA 3.1, and an inactivated 〈em〉Vibrio anguillarum〈/em〉 vaccine developed in our previous work was chosen for determining the efficiency of the CpG-riched plasmids (pCpG) as an adjuvant. Results showed that pCpG we designed can offer higher immunoprotection with the vaccine. Interestingly, even below the minimum immune dosage of the vaccine, a high RPS of 84% was observed once the vaccine was administrated with the pCpG. Serum specific antibody titer, superoxide dismutase and total protein were enhanced and some immune genes related to both innate and adaptive immune response were upregulated, implying an effective auxiliary function of the pCpG. Totally, our study suggested that the pCpG is a potential and available adjuvant for turbot vaccine development.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Guangben Wei, Shuanghu Cai, Yuanzhi Wu, Shaohong Ma, Yucong Huang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉〈em〉Vibrio harveyi〈/em〉 is the pathogen causing vibriosis in marine-cultured animals, leading to massive deaths in farmed grouper around the world. It is urgent to develop an effective vaccine to prevent vibriosis. In the previous study, we developed a 〈em〉V. harveyi〈/em〉 formalin-killed cells vaccine (FKC), and sought an effective adjuvant for enhancing the immune efficacy of vaccine. In this study, we aimed to evaluate the immune responses and protective effect of FKC combined with chitosan oligosaccharide (COS) or 〈em〉Astragalus〈/em〉 polysaccharides (APS) in the pearl gentian grouper〈em〉♀Epinephelus fuscoguttatus × ♂E. lanceolatus〈/em〉. The results indicated the vaccine triggered a remarkably higher expression levels of IL-1β, IL-16, TNF-α, MHC-Iα and IgM in the kidney and spleen of groupers post-vaccination. Antibody titers, lysozyme, catalase, superoxide dismutase and total protein were significantly elevated in the vaccinated fish compared with those in the control. The experimental groupers were challenged intraperitoneally by 〈em〉V. harveyi〈/em〉 at 35 d post-vaccination, and the relative percentage of survival (RPS) of group FKC + COS, FKC + APS, COS, APS and FKC were 80%, 72%, 52%, 47% and 55%, respectively. These results demonstrated COS and APS was the potential adjuvants for FKC against 〈em〉V. harveyi〈/em〉 in aquaculture.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Chin-Cheng Chien, Tzu-Yung Lin, Chia-Chun Chi, Chun-Hung Liu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The purpose of this study was to profile the mechanisms of action of probiotic, 〈em〉Bacillus subtilis〈/em〉 E20 in activating the immunity of white shrimp, 〈em〉Litopenaeus vannamei〈/em〉. Two groups of shrimp were studied. One group was fed a control diet without probiotic supplementation and the other was fed a probiotic-containing diet at a level of 10〈sup〉9〈/sup〉 cfu kg diet〈sup〉−1〈/sup〉. After the 8-week feeding regimen, the metabolite composition in the hepatopancreas of shrimp were investigated using 〈sup〉1〈/sup〉H nuclear magnetic resonance (〈sup〉1〈/sup〉H NMR) based metabolomic analysis. Results from the 〈sup〉1〈/sup〉H NMR analysis revealed that 16 hepatopancreatic metabolites were matched and identified among groups, of which 2 metabolites, creatinine and glutamine were significantly higher in probiotic group than in the control group. This result was confirmed by the reverse-phase high-performance liquid chromatography (RP-HPLC) and spectrophotometric analysis. Transcriptome analysis indicated the expressions of 10 genes associated with antioxidant enzymes, pattern recognition proteins and antimicrobial molecules, more active expression in the shrimp fed a diet supplemented with probiotic as compared to that of shrimp in control. In addition, the expressions of 4 genes involved with hexosamine biosynthesis pathway (HBP) and UDP-N-acetylglucosamine-peptide N-acetylglucosaminyltransferase for protein 〈em〉O-〈/em〉glycosylation were also higher in hepatopancreas of probiotic-treated shrimp than in shrimp fed a control diet. Western blot and enzyme-linked immunosorbent assay showed that heat shock factor 1, heat shock protein 70, and protein 〈em〉O〈/em〉-glycosylation in hepatopancreas were higher in probiotic group than the control group. These findings suggest that probiotic, 〈em〉B. subtilis〈/em〉 E20 promotes the digestibility of glutamine in the diet, and that the increased glutamine in shrimp can be used as fuel for immune cells or may be used to regulate immune molecule expressions and protein 〈em〉O〈/em〉-glycosylation via the HBP to increase protein 〈em〉O〈/em〉-glycosylation, thereby improving the health of shrimp.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Hourong Liu, Chengwen Song, Junhao Ning, Yuan Liu, Zhaoxia Cui〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Variable lymphocyte receptors (VLRs) play an important role via their antigen-special reorganization in jawless vertebrates (agnathans) adaptive immune response. In the present study, the open reading frame (ORF) of 〈em〉Eriocheir sinensis〈/em〉 VLRA (designated as EsVLRA) was identified. EsVLRA comprised a 799-amino-acid polypeptide with one LRR_NT domain, thirteen LRR domains and one LRR_CT domain, which showed a high domain consistency of the VLR genes in lamprey (〈em〉Petromyzon marinus〈/em〉). The transcript of EsVLRA was detected in all examined tissues with the highest level detected in hepatopancreas. Notably, the expression of EsVLRA in hepatopancreas, gonads, gill and intestine of male crabs was significantly higher than that in females. The recombinant EsVLRA exhibited strong bacteria-binding activity rather than antibacterial activity, suggesting its crucial role in immune recognition. Furthermore, 6 h earlier response and a significantly higher peak of EsVLRA mRNA expression was observed after challenge with live 〈em〉Vibrio parahaemolyticus〈/em〉 (240.6-fold, 〈em〉P〈/em〉 〈 0.01, crabs receive secondary challenge after 〈em〉V. parahaemolyticus〈/em〉 vaccine to the carbs only receive twice PBS injection, N = 6), compared with those only received first injection with formalin-inactivated 〈em〉V. parahaemolyticus〈/em〉 (39.7-fold, 〈em〉P〈/em〉 〈 0.01, challenge 6 h to vaccination 12 h). The findings of this study together demonstrated that EsVLRA plays an important role in the immune system of 〈em〉E. sinensis〈/em〉, serving as a pattern recognition receptor and involving in the immune priming.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Iris Barjhoux, Damien Rioult, Alain Geffard, Melissa Palos Ladeiro〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Immunotoxicity analysis receives a strong interest in environmental 〈em〉a priori〈/em〉 and 〈em〉a posteriori〈/em〉 risk assessment procedures considering the direct involvement of the immune system in the health status of organisms, populations and thus ecosystems. The freshwater mussel 〈em〉Dreissena polymorpha〈/em〉 is an invasive species widely used in ecotoxicology studies and biomonitoring surveys to evaluate the impacts of contaminants on aquatic fauna. Bivalve hemocytes are the immunocompetent cells circulating in the open circulatory system of the organism. However, there is nowadays no consensus on a protocol to evaluate the immunocompetent state of this particular cell type using flow cytometry. Wild species such as 〈em〉D. polymorpha〈/em〉 present several technical barriers complicating their analyze including (i) the quality and the purity of the hemolymph sample, (ii) the controversial characterization of hemocyte subpopulations and their diversity, (iii) the quantity of biological material, and (iv) the high inter-individual variability of hemocyte responses. The present work proposes several technical and analytical improvements to control the above-mentioned issues. The inclusion of sedimentation and cell detachment steps in the pre-analytical phase of the protocol substantially ameliorate the quality of the hemolymph sample as well as the accuracy of the cytometric measurements, by selecting the analyzed cells on their adhesion ability and by increasing the concentration of the analyzed events. The development of an effective triple-labeling procedure including the cellular probe Hoechst® 33342, the membrane impermeant dye propidium iodide and yellow-green fluorescent microspheres allowed the simultaneous analysis of cytotoxicity and phagocytosis activity in hemocytes. It also significantly enhanced the accuracy of hemocyte endpoint measurements by eliminating non-target events from the analysis and allowing relevant gating strategies. Finally, the use of pooled samples of hemolymph noticeably reduced inter-sample variability while providing more plasticity in the experimental design and improving the discriminating potency between treatments. The developed protocol is suitable for 〈em〉ex vivo〈/em〉 exposure of hemocyte in a chemical/environmental toxicity assessment as well as for 〈em〉in vivo〈/em〉 exposure in the laboratory or 〈em〉in situ〈/em〉 biomonitoring surveys with few adaptations.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S1050464819312318-fx1.jpg" width="500" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Limei Qiu, Hao Chen, Zhi Zhou, Huan Zhang, Rui Liu, Qilin Yi, Chuanyan Yang, Lei Gao, Lingling Wang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Oyster 〈em〉Crassostrea gigas〈/em〉, is considered as a useful environmental indicator since it is widely distributed along the intertidal zone whereby it tends to accumulate cadmium and is always exposed to various pathogen agents. However, its molecular responses to both cadmium and pathogen stimulation remain unclear. In the present study, transcriptome data of hemocytes from oysters were analyzed to reveal specific molecular responses of oyster to cadmium or cadmium/bacteria stimulation. A total of 21591, 22872 and 20107 genes were detected in the BLANK, Cd24h and Cd/Bac24h group, respectively. Among them, there were 685 differentially expressed genes collected in the comparison of Cd24h versus BLANK. GO analysis of these genes found that sixteen terms into the Molecular Function category displayed transporter activities, and were all over-enrichment by cadmium exposure, whereas twelve terms into Biological Process category involved mainly in metabolic process of the various cellular components and two terms into Cellular Component category were all under-enrichment. The 330 immune responsive genes were shared by two gene lists of CdBac24h versus BLANK and CdBac24h versus Cd24h, and seven out of thirty terms in GO analysis were related to the immune process. Further annotation of these genes from the KEGG database revealed fourteen pathways, including two nervous system related pathways, arachidonic acid pathway, four immune pathways, MAPK cascade and other four cell signaling pathways, and two energy related pathways. Twenty-two differentially expressed genes were identified to responsive to both cadmium exposure and bacteria stimulation, but in different expression patterns, suggesting that bilateral responsive genes, such as alkaline phosphatase and sodium and chloride-dependent glycine transporter gene, could be candidate biomarkers for early warning of cadmium pollution. The present results collectively indicated that a profound neuro-endocrine-immune regulatory network was activated in response to cadmium and bacteria stimulation in oyster 〈em〉C. gigas〈/em〉, and the expression pattern of some cadmium responsive genes may be either reversed or strengthened by bacteria stimulation. The results provide knowledge on the transcriptomic response profile of oyster after short-term cadmium exposure and bacteria stimulation, which would be useful for future studies on stress response mechanism of mollusc, and some cadmium-bacteria responsive genes may be explored as potential biomarkers for monitoring marine pollution.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Lei Liu, Yunfei Dang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Mannose-binding lectin (MBL) is a crucial pattern recognition receptor in the host innate immune system. Previously, we reported the biological function of 〈em〉Ctenopharyngodon idella〈/em〉 MBL (CiMBL) in initiating the lectin pathway of the complement system. In the present study, we further explored its biological function including the agglutinating ability, binding capacity and protective role 〈em〉in vitro〈/em〉 and 〈em〉in vivo〈/em〉. After 〈em〉Aeromonas hydrophila〈/em〉 infection, western blot analysis revealed that the CiMBL were fluctuated and expressed in the serum and major immune-related tissues. The result of quantitative PCR (qPCR) showed that the recombinant CiMBL (rCiMBL) significantly inhibited the mRNA expression of interleukin-1β (〈em〉IL-1β〈/em〉) and tumor necrosis factor-α (〈em〉TNF-α〈/em〉) in liver, spleen and hepatic cells. Due to rCiMBL bound to 〈span〉d〈/span〉-mannose, 〈span〉d〈/span〉-galactose, 〈span〉d〈/span〉-glucose, N-acetyl-〈span〉d〈/span〉-glucosamine (GlcNAc), lipopolysaccharide (LPS), peptidoglycan (PGN) and Agar in the presence of Ca〈sup〉2+〈/sup〉, herein gram-positive (〈em〉Staphylococcus aureus〈/em〉 and 〈em〉Micrococcus luteus〈/em〉) and gram-negative (〈em〉A. hydrophila〈/em〉 and 〈em〉Vibrio anguillarum〈/em〉) bacteria were agglutinated by rCiMBL in a Ca〈sup〉2+〈/sup〉-dependent manner. More importantly, rCiMBL enhanced the survival rate of grass carp following bacterial infection. Overall, the results provide an evidence that CiMBL can protect grass carp against 〈em〉A. hydrophila〈/em〉 infection in aquaculture.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Chao Zhang, Xueying Cao, Kaiqiang Wang, Xiaoling Dai, Ruidong Zhang, Zhuoxing Zhang, Xin Huang, Qian Ren〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Transcription factor activator protein 1 (AP1) plays an irreplaceable role in the response to a variety of external stimulants, such as cellar stress, bacterial and viral infections, and inflammatory cytokines. In this study, we identified a novel 〈em〉AP1〈/em〉 gene from 〈em〉Macrobrachium nipponense〈/em〉 and named it 〈em〉MnAP1〈/em〉, which has a full length of 1747 bp contains an 882 bp open reading frame, and encodes a protein with 293 amino acids. The MnAP1 protein contains Pfam and bZIP domains. 〈em〉MnAP1〈/em〉 is widely distributed in hemocytes, heart, hepatopancreas, gill, stomach, and intestinal tissues. The expression levels of 〈em〉MnAP1〈/em〉 in the gills and stomach were significantly upregulated after 〈em〉Vibrio parahaemolyticus〈/em〉 and 〈em〉Staphylococcus aureus〈/em〉 attacks. We studied the relationship between 〈em〉MnAP1〈/em〉 and the transcripts of antimicrobial peptides (AMPs) in gills through RNA interference. Interestingly, the regulatory effects of 〈em〉MnAP1〈/em〉 on the expression of different AMPs were different. We found that the expression levels of crustins, including 〈em〉Cru1〈/em〉, 〈em〉Cru3,〈/em〉 and 〈em〉Cru4〈/em〉 in the gills were evidently decreased, whereas the synthesis of 〈em〉Cru5〈/em〉 and anti-lipopolysaccharide factors (〈em〉ALF〈/em〉3 and 〈em〉ALF4〈/em〉) were obviously increased. We further explored the effect of 〈em〉MnAP1〈/em〉 on the expression of transcription factor relish from 〈em〉M. nipponense〈/em〉. The result showed that the knockdown of 〈em〉MnAP1〈/em〉 can remarkably upregulate the expression of 〈em〉MnRelish〈/em〉. Relish as a member of the nuclear factor κB family that regulates the expression of AMPs in the innate immunity of crustacean. Hence, we also detected the expression levels of 〈em〉Cru5〈/em〉, 〈em〉ALF3〈/em〉, and 〈em〉ALF4〈/em〉 in the gills of 〈em〉MnRelish〈/em〉-silenced prawns. The Data showed that the expression levels of these three AMPs were evidently reduced after 〈em〉MnRelish〈/em〉 silencing. Our results indicated that 〈em〉MnAP1〈/em〉 plays a positive role in regulating the expression of AMPs, promotes the JNK/AP1 signaling pathway, and exerts a negative regulatory effect on the synthesis of AMPs by inhibiting the transcription of NF-κB factor in the innate immunity of 〈em〉M. nipponense〈/em〉.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): Shuo Li, Gaixiang Hao, Yu Feng, Jiafang Li, Nan Wang, Jinsheng Sun〈/p〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): Chang-Sheng Zhao, Di-An Fang, Dong-Po Xu〈/p〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): Tianxu Zhang, Zhenguang Yan, Xin Zheng, Shuping Wang, Juntao Fan, Zhengtao Liu〈/p〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): Yulema Valero, Elena Chaves-Pozo, Alberto Cuesta〈/p〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): José María García Beltrán, Daniel González Silvera, Cristóbal Espinosa Ruiz, Vittorio Campo, Latifeh Chupani, Caterina Faggio, María Ángeles Esteban〈/p〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): Fei Zhu, Xiongchao Ma〈/p〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): Zhen Gan, Jun Cheng, Jing Hou, Hongli Xia, Wenjie Chen, Liqun Xia, Pin Nie, Yishan Lu〈/p〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): V. Stefi Raju, Purabi Sarkar, R. Pachaiappan, Bilal Ahamad Paray, Mohammad K. Al-Sadoon, Jesu Arockiaraj〈/p〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): Ran Li, Qinghao Meng, Jinwei Huang, Shen Wang, Jinsheng Sun〈/p〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): Ramasamy Harikrishnan, Gunapathy Devi, Bilal Ahmad Paray, Mohammad K. Al-Sadoon, Abdul Rahman Al-Mfarij, Hien Van Doan〈/p〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): Xiao-bo Yan, Xiao-hui Dong, Bei-ping Tan, Shuang Zhang, Shu-yan Chi, Qi-huiYang, Hong-yu Liu, Yuan-zhi Yang〈/p〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): Genmei Lin, Min Zheng, Dong Gao, Shizhu Li, Wenyu Fang, Jing Huang, Jingui Xie, Jingxiong Liu, Yijing Liu, Zhaohong Li, Jianguo Lu〈/p〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Jin-Nian Li, Yu-Ting Zhao, Shou-Lin Cao, Hong Wang, Jia-Jun Zhang〈/p〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Xiaohong Tan, Zhenzhu Sun, Chaoxia Ye〈/p〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): Hung Bao Tran, Putri Nurhanida Rizky, Stanley Rob McDaniel Padgett, Yen-Hung Lee, Hso-Chi Chaung, Ta-Chih Cheng〈/p〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): Jiu-Le Wang, Hai-Qing Wang, Ze-Quan Mo, Ling Zhou, Xiao-Chun Luo, An-Xing Li, Yan-Wei Li, Xue-Ming Dan〈/p〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): Keqiang Wei, Yue Wei, Changxia Song〈/p〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): T.D.W. Kasthuriarachchi, Qiang Wan, Jehee Lee〈/p〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): Ghasem Mohammadi, Ghasem Rashidian, Seyed Hossein Hoseinifar, Saeid Shahbazi Naserabad, Hien Van Doan〈/p〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): Chan Zhu, Dingji Shi, Shengyu Liao, Peimin He, Rui Jia〈/p〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): Arpita Nepal, Deanna L. Wolfson, Balpreet Singh Ahluwalia, Ingvill Jensen, Jorunn Jørgensen, Dimitar B. Iliev〈/p〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Sheng-Wei Luo, Zi-Ye Luo, Teng Yan, Kai-Kun Luo, Ping-Hui Feng, Shao-Jun Liu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Hepcidin, a multifunctional hormone oligopeptide, not only exhibits a regulatory role in iron metabolism, but also participates in the regulation of teleostean immunity. In this study, ORF sequence of WR-hepcidin was 258 bp and encoded 85 amino acid residues. Tissue-specific analysis revealed that the highest expression of WR-hepcidin was observed in liver. 〈em〉Aeromonas hydrophila〈/em〉 challenge can sharply increased WR-hepcidin mRNA expression in liver, trunk kidney and spleen. The purified WR-hepcidin fusion peptide can directly bind to 〈em〉A. hydrophila〈/em〉 and 〈em〉Streptococcus agalactiae〈/em〉, reduce the relative bacterial activity, limit bacterial growth and attenuate their dissemination to tissues 〈em〉in vivo〈/em〉. In addition, the treatment of WR-hepcidin fusion protein can diminish the production of pro-inflammatory cytokines. These results indicated that WR-hepcidin can play a negative regulatory role in bacteria-stimulated pro-inflammatory cytokines production and MyD88-IRAK4 activation.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): Khajida Ben Othmen, Walid Elfalleh, José María García Beltrán, M. Ángeles Esteban, Mansour Haddad〈/p〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): Can Zhang, Long-Feng Lu, Zhuo-Cong Li, Xiao-Yu Zhou, Yu Zhou, Dan-Dan Chen, Shun Li, Yong-An Zhang〈/p〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): Yidi Kong, Min Li, Jiaxin Tian, Linhui Zhao, Yuanhuan Kang, Lei Zhang, Guiqin Wang, Xiaofeng Shan〈/p〉

Description: 〈p〉Publication date: April 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 99〈/p〉 〈p〉Author(s): Seyed Hossein Hoseinifar, Meysam Shakouri, Hien Van Doan, Shafigh Shafiei, Morteza Yousefi, Mojtaba Raeisi, Samira Yousefi, Ramasamy Harikrishnan, Miriam Reverter〈/p〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Xing-Wei Liu, Lin Feng, Wei-Dan Jiang, Pei Wu, De-Min Yang, Ling Tang, Sheng-Yao Kuang, He-Qun Shi, Xiao-Qiu Zhou, Yang Liu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The present study evaluated the effects of (2-Carboxyethyl)dimethylsulfonium Bromide (Br-DMPT) supplementation on the intestinal immune function and potential mechanisms of on-growing grass carp (〈em〉Ctenopharyngodon idella〈/em〉) by feeding fish (initial weight 216.49 ± 0.29 g) five diets with gradational Br-DMPT (0–520 mg/kg diet) concentrations for 60 days and then infecting them with 〈em〉Aeromonas hydrophila〈/em〉 for 14 days. Our results firstly indicated that compared with the control group, appropriate Br-DMPT supplementation increased the number of beneficial bacteria 〈em〉Lactobacillus〈/em〉 and 〈em〉Bifidobacterium〈/em〉 and enteritis resistance, decreased the number of detrimental bacteria 〈em〉Aeromonas〈/em〉 and 〈em〉E. coli〈/em〉, and relieved the intestinal histopathological symptoms of fish. In addition, compared with the control group, appropriate Br-DMPT supplementation (1) increased lysozyme (LZ) and acid phosphatase (ACP) activities, as well as complement 3 (C3), C4 and immunoglobulin M (IgM) content; (2) upregulated the mRNA levels of anti-microbial substance: liver expressed anti-microbial peptide (LEAP) -2A, LEAP-2B, hepcidin, β-defensin-1 and Mucin2; (3) partially downregulated the mRNA levels of pro-inflammatory cytokines [interleukin 1β (IL-1β), IL-6, IL-8, IL-12p40, IL-15, IL-17D, tumour necrosis factor α (TNF-α) and interferon γ2 (IFN-γ2)] by inhibiting [IKKβ/IκBα/(NF-κBp65 and c-Rel)] signalling; and (4) partially upregulated the mRNA levels of anti-inflammatory cytokines [IL-4/13A, IL-10, IL-11, transforming growth factor (TGF)-β1] by activating [TOR/(S6K1 and 4E-BP)] signalling. The aforementioned results indicated that appropriate amount of Br-DMPT exerted a positive effect on the regulation of intestinal immune function in fish. Finally, based on enteritis morbidity, the IgM content and the lysozyme activity in the PI, the appropriate levels of Br-DMPT supplementation for on-growing grass carp were established as 295.43, 301.73 and 320.36 mg/kg diet, respectively.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Yan-li Su, Guang-hua Wang, Jing-jing Wang, Bing Xie, Qin-qin Gu, Dong-fang Hao, Hong-mei Liu, Min Zhang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉At present, several reports have indicated that the C-terminal peptides of tissue factor pathway inhibitor 1 (TFPI-1) were active antibacterial peptides. However, the functions of TFPI-1 C-terminal peptides in teleost are still very limited. In this study, a C-terminal peptide, TC26 (with 26 amino acids), derived from common carp (〈em〉Cyprinus carpio〈/em〉) TFPI-1, was synthesized and investigated for its antibacterial spectrum, action mechanism, as well as the 〈em〉in vivo〈/em〉 effects on bacterial invasion. Our results showed that TC26 was active against Gram-positive bacteria 〈em〉Micrococcus luteus〈/em〉 and 〈em〉Staphylococcus aureus〈/em〉, as well as Gram-negative bacterium 〈em〉Vibrio vulnificus〈/em〉. TC26 treatment facilitated the bactericidal process of erythromycin by enhancing the out-membrane permeability of 〈em〉V. vulnificus〈/em〉. During the bactericidal process, TC26 killed the target bacterial cells 〈em〉Vibrio vulnificus〈/em〉, by destroying cell membrane integrity, penetrating into the cytoplasm and inducing degradation of genomic DNA and total RNA. 〈em〉In vivo〈/em〉 study showed that administration of turbot with TC26 before bacterial infection significantly reduced pathogen dissemination and replication in tissues. These results indicated that TC26 is a novel and active antibacterial peptide and may play a vital role in fighting pathogenic infection in aquaculture.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Yingying Zhao, Xiaochen Zhu, Zhibin Han, Yazhao Zhang, Tengfei Dong, Yingdong Li, Jing Dong, Hua Wei, Xiaodong Li〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Chinese mitten crab 〈em〉Eriocheir sinensis〈/em〉 is probably the most important freshwater cultured crab in China. A tiny minority of brownish-orange individuals have been discovered in the long period of artificial breeding history of 〈em〉E. sinensis〈/em〉s. Those mutants are usually accompanied with slow growth rate, low molting frequency and poor survival rate, which may be the results of growth defects and immunodeficiency. To better understand the relationship between body color determination and the immune system as well as the related genes expression in 〈em〉E. sinensis〈/em〉s, we performed the whole-body transcriptome analysis in different color of first stage zoea (ZI) larvae using next-generation sequencing (NGS) technology. We randomly assembled 175.40 and 177.52million clean reads from the wild and mutant ZIs, respectively. Finally, we identified 7153 differentially expressed genes (DEGs) (〈em〉p〈/em〉 〈 0.05), with 5194 up-regulated and 1959 down-regulated. A total of 13 KEGG pathways related to immune system were detected among 248 pathways. Except the first whole-body RNA sequencing of color-specific transcriptomes for 〈em〉E. sinensis〈/em〉, this study will offer a better understanding of the underlying molecular mechanisms of interaction between color determination and the immune system.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Shu-wen He, Xue Du, Guang-hua Wang, Jing-jing Wang, Bing Xie, Qin-qin Gu, Min Zhang, Han-jie Gu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Cathepsin K belongs to the family of cysteine cathepsins. It is well known that the cysteine cathepsins participate in various physiological processes and host immune defense in mammals. However, in teleost fish, the function of cathepsin K is very limited. In the present study, a cathepsin K homologue (SsCTSK) from the teleost black rockfish (〈em〉Sebastes schlegelii〈/em〉) was identified and examined at expression and functional levels. In silico analysis showed that three domains, including signal peptide, cathepsin propeptide inhibitor I29 domain, and functional domain Pept_C1, are existed in SsCTSK. SsCTSK also possesses a peptidase domain with three catalytically essential residues (Cys25, His162 and Asn183). Phylogenetic profiling indicated that SsCTSK was evolutionally close to the cathepsin K of other teleost fish. Expression of 〈em〉SsCTSK〈/em〉 occurred in multiple tissues and was induced by bacterial infection. Purified recombinant SsCTSK (rSsCTSK) exhibited apparent maximal peptidase activity at 45 °C, and its enzymatic activity was remarkably declined in the presence of the cathepsin inhibitor E−64. Moreover, rSsCTSK possesses the ability to bind with PAMPs and bacteria. Finally, knockdown of 〈em〉SsCTSK〈/em〉 expression facilitated bacterial invasion in black rockfish. Collectively, these results indicated that SsCTSK functions as a cysteine protease and may serves as a target for pathogen manipulation of host defense system.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Xinghai Zhu, Huan Liao, Zujing Yang, Cheng Peng, Wei Lu, Qiang Xing, Xiaoting Huang, Jingjie Hu, Zhenmin Bao〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs), are crucial sensors with a conserved structure in cytoplasm, inducing the production of cytokines, chemokines and host restriction factors which mediate a variety of intracellular activities to interfere with distinct PAMPs (pathogen-associated molecular patterns) for eliminating pathogens in innate immune system. Although 〈em〉RLR〈/em〉 genes have been investigated in most vertebrates and some invertebrates, the systematic identification and characterization of 〈em〉RLR〈/em〉 genes have not been reported in scallops. In this study, four 〈em〉RLR〈/em〉 genes (〈em〉PY-10413.4〈/em〉, 〈em〉PY-10413.5〈/em〉, 〈em〉PY-443.7〈/em〉 and 〈em〉PY-443.8〈/em〉, designated 〈em〉PyRLRs〈/em〉) were identified in Yesso scallop (〈em〉Patinopecten yessoensis〈/em〉) through whole-genome scanning through 〈em〉in silico〈/em〉 analysis, including two pairs of tandem duplicate genes located on the same scaffold (〈em〉PY-10413.4〈/em〉 and 〈em〉PY-10413.5〈/em〉, 〈em〉PY-443.7〈/em〉 and 〈em〉PY-443.8〈/em〉, respectively). Phylogenetic and protein structural analyses were performed to determine the identities and evolutionary relationships of these genes. The expression profiles of 〈em〉PyRLRs〈/em〉 were determined in all developmental stages, in healthy adult tissues, and in mantles that simulated ocean acidification (OA) exposure (pH = 6.5 and 7.5) at different time points (3, 6, 12 and 24 h). Spatiotemporal expression patterns suggested the functional roles of 〈em〉PyRLRs〈/em〉 in all stages of development and growth of the scallop. Regulation expressions revealed 〈em〉PY-10413.4〈/em〉 and 〈em〉PY-10413.5〈/em〉 with one or two CARD(s) (caspase activation and recruitment domain) were up-regulated expressed at most time points, whereas 〈em〉PY-443.8〈/em〉 and 〈em〉PY-10413.4〈/em〉 without CARD were significantly down-regulated at each time points, suggesting functional differentiations in the two pairs of 〈em〉PyRLRs〈/em〉 based on the structural differences in response to OA. Collectively, this study demonstrated gene duplication of RLR family genes and provide primary analysis for versatile roles in the response of the bivalve innate immune system to OA challenge.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Mingming Zhang, Jiawei Wang, Qirui Sun, Hu Zhang, Peng Chen, Qiang Li, Yinan Wang, Guo Qiao〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Along with rapid offshore and onshore wind power development in modern society, extremely low frequency electromagnetic fields (ELF-EMF) is produced extensively in the habits of aquatic organisms. However, the biological effects of ELF-EMF on aquatic organisms are almost sparse. In this study, 〈em〉Onchidium struma〈/em〉 without shell was chosen to aim whether ELF-EMF can elicit immune response of mollusk based on immune-related enzyme activities and gene expression through high-throughput transcriptome sequencing. Three experimental groups, i.e. ELF-EMF unexposed control group (C), ELF-EMF (50 Hz, 100 μT) exposed E1 group, and ELF-EMF (50 Hz, 500 μT) exposed E2 group, were set, and coelomocytes were collected to analyze. The results showed that total coelomocyte and spherulocyte density in E1 group increased significantly compared to groups C and E2 (〈em〉P〈/em〉 〈 0.05). There were no significant differences on amoebocyte and chromatocyte density among groups C, E1 and E2. ELF-EMF exposure could significantly increase immune-related enzyme activities in coelomic fluid of 〈em〉O. struma〈/em〉, including acidic phosphatase, alkaline phosphatase, antioxidative capacity, catalase, superoxide dismutase, and polyphenol oxidase (〈em〉P〈/em〉 〈 0.05). A total of 54.32 Mb and 55.27 Mb raw reads with average length of 1520 bp were obtained from coelomocytes of 〈em〉O. struma〈/em〉 in unexposed and exposed groups, respectively. There were 341 differentially expressed genes (DGEs) between unexposed and exposed groups, including 209 up-regulated and 132 down-regulated unigenes. All the DGEs were allocated to 14 Kyoto Encyclopedia of Genes and Genomes pathways, and five pathways were associated with immune response, including TLR/TNF/NOD-like receptor/MAPK/Fc epsilon RI signaling pathways. Altogether, short-term (to one week) exposure of 〈em〉O. struma〈/em〉 to lower luxy density ELF-EMF (〈500 μT) could elicit the immune response, and antioxidant system is recommended as indicators of immunological effects. Hopefully, this study will further provide insights into exploring biomarker for evaluation of the effect of ELF-EMF exposure on aquatic organisms regarding to field density, frequency and exposure duration, and provide good guidance for exploitation and utilization of renewable energy.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 11 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology〈/p〉 〈p〉Author(s): Libing Zheng, Jiaying Qiu, Jia Chen, Wei-qiang Zheng, Ying Pan〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉〈em〉Cryptocaryon irritans〈/em〉 infection could cause huge economic losses to the marine fish industry. 〈em〉Larimichthys crocea〈/em〉, a special economic species in China, suffered from the threat of serious infection, and 〈em〉L. crocea〈/em〉 could enhance the level of piscidin 5-like to defense against the infection. This study set out to observe the main histopathological changes of some key tissues caused by infection, and determineed how an ectoparasite affected the expression of piscidin-5 like in its hosts. Pathological changes and immune response were assessed using histological and in situ hybridization (ISH) technologies. The infection induced inflammation occurring, especially in the gill where epithelium cells swell, hyperplasia, necrosis shedding adjacent to the parasites attachment sites. Infected hepatic cells grown big vacuoles in the cytoplasm. The boundary between red pulp and white pulp turned indistinct, splenic corpuscle lost the normal structure, the number and size of melano-macrophage centers increased apparently in the infected spleen. The whole structure of head kidney became loose. Immunostaining with RNA probes against piscidin 5-like showed subpopulations of mast cells (MCs) were positive. Piscidin 5-like-positive MCs existed mainly in the head kidney where they distributed around melano-macrophage center, followed in the gill located at different positions they also distributed in the margin of spleen, and randomly and sparsely existed in the liver. After being infected by 〈em〉C. irritans〈/em〉, the gill arch arose positive MCs groups, and they also migrated to spleen, while the positive staining deepen in other detected tissues. Therefore, organism enhanced the expression level through improving expression ability of positive MCs, or increasing the number of positive MCs.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Ali Khani Oushani, Mehdi Soltani, Najmeh Sheikhzadeh, Mehdi Shamsaie Mehrgan, Houman Rajabi Islami〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉In this study, rainbow trout 〈em〉Oncorhynchus mykiss〈/em〉 weighing 27.75 ± 0.34 g were orally subjected to eight experimental diets each in three replicates containing varying amounts of chitosan and nano-chitosan (0.05, 0.5 and 5 g kg〈sup〉−1〈/sup〉) loaded in clinoptilolite (14.28 g kg〈sup〉−1〈/sup〉) for 70 days; and the growth and immune responses were evaluated. Results showed that growth parameters in fish fed diets chit + clin2, chit + clin3, nchit + clin1, nchit + clin2 and nchit + clin3 were significantly higher than in fish fed the control diet. All feeds, except chit + clin3, and nchit + clin3, significantly increased the total protein level. Feeds containing chit + clin2, nchit + clin1, and nchit + clin2 significantly elevated serum lysozyme activity compared with the control group. All treatments, except chit + clin3, and nchit + clin3 exhibited higher serum immunoglobulin (Ig) level than control one. In contrast, diet nchit + clin1 significantly unregulated the expression of Ig M gene in fish head-kidney compared to other groups. Additionally, all feeds, except clinoptilolite, and nchit + clin3, significantly improved the serum complement activity. Diets chit + clin2, nchit + clin1, and nchit + clin2 also significantly elevated antibacterial activity against 〈em〉Yersinia ruckeri〈/em〉 compared with the control diet. Expression of inducible nitric oxide synthase (iNOS) gene in fish fed diets clinoptilolite, chit + clin1, chit + clin3, nchit + clin1, nchit + clin2, and nchit + clin3 was significantly higher than the control diet. All diets, except clinoptilolite, increased IL-1β gene expression compared to the control group. Present results suggest that diets supplemented with nchit + clin, especially at 0.05 g kg〈sup〉−1〈/sup〉 nano-chitosan inclusion, could improve growth performance and immune parameters of rainbow trout.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Naphakorn Srichaiyo, Sudaporn Tongsiri, Seyed Hossein Hoseinifar, Mahmoud A.O. Dawood, Maria Ángeles Esteban, Einar Ringø, Hien Van Doan〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The present study addressed the possible effects of fishwort 〈em〉(Houttuynia cordata)〈/em〉 powder (FWP) on Nile tilapia's skin mucus parameter, serum immune response, and growth performance. Three hundred twenty tilapia fingerlings (average weight of 39.06 ± 0.16 g) were divided into four treatments and fed four levels of FWP; 0, 5, 10, and 20 g kg〈sup〉−1〈/sup〉 for 72 days. Completed randomised design of the four replications was applied and revealed that fish fed FWP significantly improved skin mucus lysozyme activity (SMLA). The highest value (〈em〉P〈/em〉 〈 0.05) was recorded in fish fed 10 g kg〈sup〉−1〈/sup〉 FWP. However, no significant difference in SMLA was observed by feeding the fish 5 and 20 g kg〈sup〉−1〈/sup〉 FWP. Significant (〈em〉P〈/em〉 〈 0.05) enhanced skin mucus peroxidase activity (SMPA) was observed in fish fed 10 g kg〈sup〉−1〈/sup〉 FWP, but no significant difference in SMPA was detected between FWP supplemented diets (5 and 20 g kg〈sup〉−1〈/sup〉 FWP) and the control group. Regarding serum immunity, dietary administration of FWP showed significantly (〈em〉P〈/em〉 〈 0.05) improved serum lysozyme, peroxidase, alternative complement (ACH50), and phagocytosis 〈em〉vs〈/em〉. the control. The highest values of serum immunity (〈em〉P〈/em〉 〈 0.05) were recorded in fish fed 10 g kg〈sup〉−1〈/sup〉 FWP. However, no significance in respiratory burst activity was observed. Similarly, no significant difference in growth performance, feed conversion ratio, and survival rate was observed in fish fed FWP compared to the control. In summary, diets supplemented with FWP (10 g kg〈sup〉−1〈/sup〉) increased the serum and mucosal immunity; however, no FWP supplementations had effects on Nile tilapia growth and survival rate.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Xifeng Qiao, Pingchao Li, Jianan He, Zeshu Yu, Jiaxing Chen, Liangge He, Xue Yu, Haoran Lin, Danqi Lu, Yong Zhang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Scavenger receptors play a central role in defending against infectious diseases in mammals. However, the function of SRECII remains unknown in teleost fish. In this study, type F scavenger receptor expressed by endothelial cells-II (SRECII) cDNA sequence was first identified from 〈em〉Epinephelus coioides〈/em〉, named 〈em〉Ec〈/em〉SRECII, which contained an N-terminal signal peptide, eight EGF/EGF-like cysteine-rich motifs and a C-terminal low-complexity region. The gene location maps revealed that 〈em〉Ec〈/em〉SRECII has the conservation of synteny among selected species. Subcellular localization showed that 〈em〉Ec〈/em〉SRECII was mainly located in the cytoplasm in HEK293T cells and GS cells. In healthy 〈em〉E. coioides〈/em〉, 〈em〉Ec〈/em〉SRECII mRNA was highly expressed in spleen, skin, gill, thymus and head kidney. The relative 〈em〉Ec〈/em〉SRECII mRNA expression after 〈em〉Vibrio parahaemolyticus〈/em〉 infection was significantly up-regulated at 12 h in spleen, head kidney and thymus, but downregulated at 1 d in skin and reduced at 3 d and 1 w in spleen. Furthermore, overexpression of 〈em〉Ec〈/em〉SRECII activated NF-κB and IFN-β signaling pathway 〈em〉in vitro〈/em〉. Taken together, these results indicated that 〈em〉Ec〈/em〉SRECII could be as the potential pathogen recognition receptor for involving in bacterial infection by regulating innate immunity responses in 〈em〉E. coioides〈/em〉.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Yuyuan Wu, Junjun He, Gaoyou Yao, Haiying Liang, Xuemin Huang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Proteins in the tumor necrosis factor receptor (TNFR) superfamily play significant roles in many physiological and pathological events, such as inflammation, apoptosis, autoimmunity, and organogenesis. Here, two TNFR gene homologs (〈em〉PmTNFR1〈/em〉 and 〈em〉PmTNFR5〈/em〉) were identified in the pearl oyster 〈em〉Pinctada fucata martensii.〈/em〉 The predicted PmTNFR1 and PmTNFR5 protein sequences were 406 and 533 amino acids long, respectively, and both possessed motifs characteristic of the TNFR family, including a TNFR homology domain (CRD), a transmembrane domain (TM), and death domains. However, the predicted amino acid sequences of PmTNFR1 and PmTNFR5 had low identity (~16–23%) with sequences of vertebrate TNFR family proteins. Furthermore, PmTNFR5 had a death domain at the C-terminal, indicating that this protein may be a novel member of the TNFR superfamily. Constitutive 〈em〉PmTNFR1〈/em〉 and 〈em〉PmTNFR5〈/em〉 mRNA expression was detected in all six pearl oyster tissues tested, with comparatively greater transcript abundance in the hepatopancreas and gill. The gene expression levels of 〈em〉PmTNFR1〈/em〉 and 〈em〉PmTNFR5,〈/em〉 as well as those of downstream signaling molecules related to the NF-κB pathway (〈em〉RIP, TRAF2, TRAF3, IKK,〈/em〉 and 〈em〉NF-κB〈/em〉), were quantified in the gill after LPS challenge and in the hemocytes after nucleus insertion surgery using real-time PCR (qRT-PCR). We found that all genes were significantly upregulated at 6 h and 12 h post-injection, as well as at 15 d post-insertion. We used RNAi to inhibit the expression of the 〈em〉PmTNFR1〈/em〉 and 〈em〉PmTNFR5〈/em〉 genes. We then quantified the expression levels of 〈em〉PmTNFR1〈/em〉 and 〈em〉PmTNFR5〈/em〉, as well as downstream genes, using qRT-PCR. We found that RNAi inhibition of 〈em〉PmTNFR1〈/em〉 and 〈em〉PmTNFR5〈/em〉 downregulated the downstream genes (〈em〉RIP〈/em〉, 〈em〉TRAF2, TRAF3, IKK,〈/em〉 and 〈em〉NF-κB〈/em〉). Therefore, our results suggested that 〈em〉PmTNFR1〈/em〉 and 〈em〉PmTNFR5〈/em〉 mediate the NF-κB signaling pathway, and are closely related to immune defense, particularly allograft immunity, in the pearl oyster 〈em〉P. fucata martensii〈/em〉.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Qiuhua Li, Yuhong Chen, Libing Xu, Yusheng Yang, Qiao Wen, Li Gu, Jingqun Ao, Xinhua Chen〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Granulocyte colony-stimulating factor (GCSF) is a growth factor that drives the proliferation and differentiation of granulocytes and monocytes/macrophages. Currently, two copies of GCSF, named GCSFa and GCSFb, have been identified in teleost fish, but data on the functions and signal pathways of these fish GCSFs are still limited. In the present study, a GCSFa homologue (〈em〉Lc〈/em〉GCSFa) was identified from large yellow croaker (〈em〉Larimichthys crocea〈/em〉). The open reading frame (ORF) of 〈em〉Lc〈/em〉GCSFa is 636 bp long and encodes a protein of 211 amino acids (aa), with a 19-aa signal peptide and a typical IL-6 domain, conserved in fish GCSF sequences. The phylogenetic analysis showed that 〈em〉Lc〈/em〉GCSFa clustered with other fish GCSFa homologues. 〈em〉Lc〈/em〉GCSFa was constitutively expressed in all tissues tested and significantly up-regulated in head kidney and spleen by 〈em〉Vibrio alginolyticus〈/em〉 or poly(I:C). 〈em〉Lc〈/em〉GCSFa transcripts were also detected in primary head kidney leucocytes (PKL), primary head kidney macrophages (PKM), and primary head kidney granulocytes (PKG), and significantly up-regulated in PKL and PKG by LPS or poly(I:C)〈em〉.〈/em〉 These data indicated that 〈em〉Lc〈/em〉GCSFa may be involved in the immune responses induced by bacterium and virus. The recombinant 〈em〉Lc〈/em〉GCSFa protein (r〈em〉Lc〈/em〉GCSFa) produced in 〈em〉Pichia pastoris〈/em〉 promoted the proliferation of PKL both 〈em〉in vivo〈/em〉 and 〈em〉in vitro〈/em〉. Furthermore, r〈em〉Lc〈/em〉GCSFa significantly increased both transcription and phosphorylation levels of the signal transducers and activators of transcription (STAT) proteins (〈em〉Lc〈/em〉STAT3 and 〈em〉Lc〈/em〉STAT5) in PKL, which are required for the GCSF-dependent proliferation. These results showed that 〈em〉Lc〈/em〉GCSFa may promote the proliferation of PKL via the activation of 〈em〉Lc〈/em〉STAT3 and 〈em〉Lc〈/em〉STAT5, suggesting a conserved role across vertebrate GCSFs.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Yun Sun, Susu Ding, Mingwang He, Anzhu Liu, Hao Long, Weiliang Guo, Zhenjie Cao, Zhenyu Xie, Yongcan Zhou〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉〈em〉Vibrio harveyi〈/em〉, a severe pathogen infects different kinds of sea animals, causes huge economic loss in aquaculture industry. In order to control the Vibriosis disease caused mainly by 〈em〉V〈/em〉. 〈em〉harveyi〈/em〉 and other 〈em〉Vibrio spp〈/em〉., the best solution lies in developing corresponding efficient vaccines. In this study, we have cloned and analysed a putative antigen TssJ from the T6SS of 〈em〉V〈/em〉. 〈em〉harveyi〈/em〉, which has the potential as a vaccine against infection. The sequence analysis and western blotting experiments indicated that TssJ anchored in outer membrane and there were several antigenic determinants existed on its extracellular region. Two forms of universal vaccines, subunit vaccine and DNA vaccine, were developed based on TssJ and applied in 〈em〉Trachinotus ovatus〈/em〉. The results showed that both of the two vaccines could generate a moderate protection in fish against 〈em〉V. harveyi〈/em〉. The relative percentage survival (RPS) of subunit vaccine and DNA vaccine were 52.39% and 69.11%, respectively. Immunological analysis showed both subunit vaccine and DNA vaccine enhanced acid phosphatase, alkaline phosphatase, superoxide dismutase, and lysozyme activities. Specific serum antibodies against TssJ in the fish vaccinated with subunit vaccine was much higher than that in the DNA vaccine group. Several immune-related genes, i.e., IL10, C3, MHC Iα, MHC IIα, and IgM, were induced both by the two forms of vaccines. TNFα and Mx were only upregulated in the DNA vaccine group. However, the induction levels of these genes induced by DNA vaccine were higher than subunit vaccine. All these findings suggested that TssJ from 〈em〉V. harveyi〈/em〉 had a potential application value in vaccine industry.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Yuan Ren, Jialin Zhang, Yinan Wang, Jing Chen, Chunlei Liang, Ruijun Li, Qiang Li〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Coelomic fluid contains a population of coelomocytes, enzymes, nutrients and kinds of molecules that could be essential for 〈em〉Apostichopus japonicus〈/em〉 live. The coelom and polian vesicle are the main tissues that hold the most coelomic fluid in the animal, but whether there exists any immunological difference of the coelomic fluid from the two tissues remains unknown. In this study, we first extracted the coelomic fluid both from the coelom and polian vesicle, and compared their non-specific immune factors. It was found that the ACP and AKP activities in the polian vesicle were significantly higher than those in the coelom, but it was contrary for the SOD and CAT. Meanwhile, the expression levels of several immune-related genes including AjC3-2, AjMKK3/6, AjTLR3 and AjToll in the polian vesicle were significantly lower than those in the coelom. Besides, the early changes of non-specific immune factors were further monitored after eviscerated. During 7 days post evisceration, the immunoenzymes activities of ACP, AKP, SOD and CAT were decreased first and then recovered gradually in the coelomic fluid from the coelom. In the polian vesicle, the ACP and AKP activities showed a similar trend with the coelom, while the SOD and CAT activities showed a transitory increase during 2 h post evisceration (hpe) to 12 hpe. Moreover, the expression profiles of the immune-related genes in the coelom reached the peak at 3 days post evisceration (dpe), while their expression levels in the polian vesicle reached the peak at 7 dpe. All the results suggested that the immunocompetence of coelomic fluid differed in the coelom and polian vesicle, and thus may exert their respective immunological functions. It was likely that the respond speed in the coelom would be faster than that in the polian vesicle after evisceration. Our data will provide a basis for better understanding of the immune defense mechanism of 〈em〉A. japonicus〈/em〉.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Yuan Fang, Xiao-yan Xu, Yubang Shen, Jiale Li〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉〈em〉Ctenopharyngodon idella〈/em〉 growth arrest and DNA damage-inducible 45 ab (〈em〉CiGadd45ab〈/em〉) is a subtype of the Gadd45a gene of the Gadd45 family in grass carp. There is increasing evidence that microRNAs (miRNAs) are involved in the regulation of inflammatory and apoptotic responses. However, little is known about the regulatory effects of miRNAs on 〈em〉CiGadd45ab〈/em〉 expression. In the present study, 〈em〉CiGadd45ab〈/em〉 was identified as a target gene of miR-23a-3p and miR-23a-5p, based on miRNA expression profiling and a dual-luciferase reporter assay. In addition, miR-23a-3p and miR-23a-5p were both confirmed to be involved in the inflammatory response following infection with 〈em〉Aeromonas hydrophila〈/em〉 by targeting 〈em〉CiGadd45ab〈/em〉. Transfection with miR-23a-3p and miR-23a-5p mimics and inhibitor altered proinflammatory gene expression and apoptosis rate, thereby suggesting that miRNAs regulate immune response and anti-apoptosis by targeting 〈em〉CiGadd45ab〈/em〉 in grass carp. Our results provide a theoretical basis for exploring the molecular mechanisms by which miR-23a-3p and miR-23a-5p target CiGadd45ab to regulate inflammation and apoptosis against bacterial infection in grass carp.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): C. Flores-Kossack, R. Montero, B. Köllner, K. Maisey〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉 〈p〉In Chile, the salmon and trout farmed fishing industries have rapidly grown during the last years, becoming one of the most important economic sources for the country. However, infectious diseases caused by bacteria, virus, mycoses and parasites, result in losses of up to 700 million dollars per year for the Chilean aquaculture production with the consequent increase of antibiotic and antiparasitic usage. After 30 years of its first appearance, the main salmon health problem is still the salmonid rickettsial septicaemia (SRS), which together with other disease outbreaks, reveal that vaccines do not provide acceptable levels of long-lasting immune protection in the field. On the other hand, due to the large dependence of the industry on salmonids production, the Chilean government promoted the Aquaculture diversification program by 2009, which includes new species such as 〈em〉Merluccius australis〈/em〉, 〈em〉Cilus gilberti〈/em〉 and 〈em〉Genypterus chilensis〈/em〉, however, specific research regarding the immune system and vaccine development are issues that still need to be addressed and must be considered as important as the farm production technologies for new fish species. Based on the experience acquired from the salmonid fish farming, should be mandatory an effort to study the immune system of the new species to develop knowledge for vaccination approaches, aiming to protect these aquaculture species before diseases outbreaks may occur.〈/p〉 〈p〉This review focuses on the current status of the Chilean aquaculture industry, the challenges related to emerging and re-emerging microbial pathogens on salmonid fish farming, and the resulting needs in the development of immune protection by rational designed vaccines. We also discussed about what we have learn from 25 years of salmonid researches and what can be applied to the new Chilean farmed species on immunology and vaccinology.〈/p〉 〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Seong Don Hwang, Kwang-Min Choi, Jee Youn Hwang, Mun-Gyeong Kwon, Ji-Min Jeong, Jung Soo Seo, Bo-Yeong Jee, Chan-Il Park〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Calpains (〈em〉CAPNs〈/em〉) belong to the papain superfamily of cysteine proteases, and they are calcium-dependent cytoplasmic cysteine proteases that regulate a variety of physiological processes. We obtained the sequence of 〈em〉CAPN3〈/em〉 from an NGS-based analysis of 〈em〉Pagrus major〈/em〉 (〈em〉PmCAPN3〈/em〉) and confirmed the conserved molecular biological properties in the predicted amino acid sequence. The amino acid sequence and predicted domains of 〈em〉CAPN3〈/em〉 were found to be highly conserved in all of the examined species, and one catalytic domain and four calcium binding sites were identified. In healthy 〈em〉P. major〈/em〉, the 〈em〉PmCAPN3〈/em〉 mRNA was most abundantly expressed in the muscle and skin, and ubiquitously expressed in the other tissues used in the experiment. After artificial infections with fish pathogens, significant changes in its expression levels were found in immune-related tissues, most of showed upregulation. In particular, the highest level of expression was found in the liver, a tissue associated with protease activity. Taken together, these results suggest a physiological activity for 〈em〉PmCAPN3〈/em〉 in 〈em〉P. major〈/em〉 and reveal functional possibilities that have not yet been reported in the immune system.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Yanping Qin, Yuehuan Zhang, Xingyou Li, Zohaib Noor, Jun Li, Zihua Zhou, Duo Xu, Zehui Zhao, Zhiming Xiang, Ziniu Yu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Caspase 3 plays an important role in apoptotic pathways and contributes to maintaining the homeostasis of the immune system in organisms. The structure, functions, and characteristics of caspase 3 have been extensively investigated in many species, but the research is scarce when it comes to bivalves, particularly oysters. In this study, we identified and cloned a previously unknown caspase 3 gene, named 〈em〉Ch〈/em〉Cas 3, in 〈em〉C. hongkongensis〈/em〉. The full-length cDNA of 〈em〉Ch〈/em〉Cas 3 was 1562 bp and included a 175 bp 5′-untranslated region (UTR), a 141 bp 3′-UTR and a 1245 bp open reading frame (ORF) that encoded a polypeptide of 415 amino acids. Similar to caspase 3 in other species, 〈em〉Ch〈/em〉Cas 3 has a pro-domain, a conserved cysteine active site, a large p20 subunit and a small p10 subunit. Our findings demonstrated the expression of 〈em〉Ch〈/em〉Cas 3 in all the eight tissues via tissue-specific expression assays with the highest expression in haemocytes. 〈em〉Ch〈/em〉Cas 3 was confirmed to be expressed throughout the larval development stages, and fluorescence from pEGFP-N1-〈em〉Ch〈/em〉Cas 3 was found to be distributed throughout the entire HEK293T cell. In addition, the relative expression of 〈em〉Ch〈/em〉Cas 3 significantly enhanced in hemocytes post bacterial stimulation, and overexpression of 〈em〉Ch〈/em〉Cas 3 led to upregulation of the transcriptional activity of NF-κB and p53 reporter genes in HEK293T cells, which indicated that it was involved in innate immune responses. Finally, the apoptosis rate of the haemocytes declined considerably compared with that of the control group after the expression of 〈em〉Ch〈/em〉Cas 3 was successfully silenced by dsRNA, corroborating its sentinel role in apoptosis. This study provides comprehensive underpinning evidences, affirming caspase 3 crucial role against bacterial infection and apoptosis in 〈em〉C. hongkongensis〈/em〉.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Zhuojun Han, Yi Zhou, Xiaojin Zhang, Jinpeng Yan, Jun Xiao, Yongju Luo, Huifang Zheng, Huan Zhong〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Ghrelin is a peptide hormone secreted by gastrointestinal tract which regulates multiple physiological processes such as appetite, metabolism, growth and gonad development in fish. In the present study, the effects of ghrelin on hybrid tilapia infected with 〈em〉Aeromonas hydrophila〈/em〉 are elucidated. Juvenile hybrid tilapia fish (20.0 ± 5.0 g) were intraperitoneally injected with 0, 0.1, 1.0, or 10.0 ng/g ghrelin/body weight synthetic ghrelin alone or in combination with 〈em〉A. hydrophila〈/em〉 (0.5 × 10〈sup〉6〈/sup〉 CFU). At 10 days post treatment, the survival rate in the group that received 1.0 ng/g ghrelin/body weight ghrelin in combination with 〈em〉A. hydrophila〈/em〉 was higher (66.66%) than that of the Ah group (13.33%) that received 〈em〉A. hydrophila〈/em〉 alone. In tilapia that received ghrelin injections, reactive oxygen species (ROS) levels tended to increase at 5 h, while injection of 10.0 ng/g ghrelin/body weight ghrelin resulted in a significant decrease in ROS levels at 10 h. No changes in serum immune or antioxidant-related indicators were observed in fish injected with 〈em〉A. hydrophila〈/em〉 compared to controls. However, ghrelin injection decreased Albumin (ALB), glutathione peroxidase (GSH-Px), lysozyme (LZM) and superoxide dismutase (SOD). Histological analysis showed that ghrelin injection alleviated the pathological changes in liver and spleen caused by 〈em〉A. hydrophila〈/em〉 infection. Overall, the expression of HSP70, IL-1β, and TGF-β in the liver tended to upregulate compared to the control. In the kidney, HSP70, IL-1β and TGF-β levels were increased, and TNF-α expression levels were decreased compared to the control. The HSP70 level in the spleen was decreased, and IL-1β, TGF-β, and TNF-α were expressed at significantly higher levels in the spleen in the tilapia that received ghrelin injections. Taken together, our results indicate that injection with 1.0 ng/g ghrelin/body weight ghrelin may effectively protect juvenile hybrid tilapia against 〈em〉A. hydrophila〈/em〉 infection by improving hematological indicators, maintaining normal histology and regulating cytokine gene expression.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Chin-Chyuan Chang, Hsin-Wei Kuo, Chang-Chi Liu, Winton Cheng〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Tyramine (TA), a biogenic monoamine, plays various important physiological roles including immunological regulation in invertebrates. In this study, the effects of TA on the regulation of immune resistance, carbohydrate metabolism and biogenic monoamine, as well as its signaling pathway in 〈em〉Macrobrachium rosenbergii〈/em〉 were determined. Results showed that total haemocyte count, hyaline cells, semigranular cells, and phenoloxidase activity per 50 μL of haemolymph and per granulocyte (the sum of semigranular and granular cells) at 0.5 h as well as phagocytic activity and clearance efficiency to 〈em〉Lactococcus garvieae〈/em〉 at 1 h of prawn injected with TA at 1 nmol prawn〈sup〉−1〈/sup〉 significantly increased, but the significantly decreased plasma lysozyme activity, phagocytic activity, clearance efficiency, and haemolymph glucose and dopamine were observed in prawn injected with TA at 10 nmol prawn〈sup〉−1〈/sup〉 for 0.5 h. Respiratory bursts and haemolymph lactate in two TA-injection treatments at 0.5 h and 0.5–1 h, respectively, were significantly higher than those of the saline control, and in addition, TA depressed dopamine release in a dose-dependent manner after 0.5 h of TA injection. All the examined parameters returned to control levels after prawn injected with TA for 2 h. The inhibited effect of TA (at 10 nmol prawn〈sup〉−1〈/sup〉 injection) on the phagocytic activity and clearance efficiency to pathogens was blocked by prazosin (an α1 adrenoceptors antagonist). For prawn received TA for 1 h then challenged with 〈em〉Lactococcus garvieae〈/em〉 at 2 × 10〈sup〉5〈/sup〉 colony-forming units prawn〈sup〉−1〈/sup〉, the survival ratio of TA 1 nmol prawn〈sup〉−1〈/sup〉-injected prawn significantly increased by 20%, compared to the saline-challenged control or TA 10 nmol prawn〈sup〉−1〈/sup〉-injected prawn after 144 h of challenge. These results suggested that the level of dopamine release suppression regulated by TA resulted in the immunoenhancing or immunosuppressive effects in prawn, and the signaling pathways of TA in mediating immune function were through octopamine (OA)/TA receptors.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 3 January 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology〈/p〉 〈p〉Author(s): Yuebo Zhao, Kaini Zheng, Fei Zhu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Troponin C (TnC) is one member of the EF-hand superfamily. In many species, this gene had been identified and related functions had been elucidated. The TnC gene was still blank in the 〈em〉Scylla paramamosain〈/em〉. We obtained the TnC gene for the first time in the 〈em〉S. paramamosain〈/em〉. And we systematically analyzed the possible role of this gene in the innate immunity of 〈em〉S. paramamosain〈/em〉 while infected with white spot syndrome virus (WSSV) or 〈em〉Vibrio alginolyticus〈/em〉. The full-length 1427 bp sequence of TnC contains a 453 bp open reading frame (ORF) for encoding a 151 amino acid protein. Detection of tissue specificity of gene expression showed that the TnC was primarily expressed in muscle tissue. The expression of TnC was successfully inhibited by RNA interference technology, and several immune genes were affected. The activity of phenoloxidase and superoxide dismutase increased, and the total hemocytes counts increased after RNAi of TnC. It was found that after infection with 〈em〉V. alginolyticus〈/em〉 and WSSV, the expression of TnC in hemocytes decreased. Infected with 〈em〉V. alginolyticus〈/em〉 and WSSV, the cumulative mortality and apoptotic rate of hemocytes increased after silencing the TnC gene. Our results indicate that TnC takes participate in the innate immunity of 〈em〉S. paramamosain〈/em〉 and may plays a different role in the antiviral and antibacterial immune response.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Junzhi Zhang, Qiangde Liu, Yuning Pang, Xiang Xu, Kun Cui, Yanjiao Zhang, Kangsen Mai, Qinghui Ai〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Apart from mitigating endoplasmic reticulum (ER) stress, vast studies have demonstrated the crucial role of inositol-requiring transmembrane kinase and endonuclease 1α (IRE1α) - spliced X-box binding protein 1 (XBP1s) signaling pathway in inflammatory response in mammals. In addition, palmitic acid (PA)-induced inflammation has been verified in large yellow croaker (〈em〉Larimichthys crocea〈/em〉). However, whether the IRE1α-XBP1s signaling pathway is involved in inflammatory response caused by PA remains poorly studied in fish. The present study was aimed at elucidating the role of the IRE1α-XBP1s signaling pathway in inflammatory response induced by PA in primary hepatocytes from large yellow croaker. In the present study, the full-length cDNA of 〈em〉ire1α〈/em〉 and 〈em〉xbp1s〈/em〉 were cloned and comprised 3793 bp and 1789 bp with an open reading frame of 3279 bp and 1170 bp, encoding 1093 and 390 amino acids, respectively. IRE1α protein possessed a protein kinase and endoribonuclease domain and XBP1s protein possessed a basic-leucine zipper domain. The IRE1α protein and XBP1s protein located to the ER membrane and nucleus respectively. The 〈em〉ire1α〈/em〉 and 〈em〉xbp1s〈/em〉 were widely transcribed in various tissues with the higher level in intestine, liver, adipose and head kidney. The ER stress-inducing agent tunicamycin (Tm) and PA treatment significantly activated the IRE1α-XBP1s signaling pathway and increased the pro-inflammatory genes expression including tumor necrosis factor α (〈em〉tnfα〈/em〉), interleukin 6 (〈em〉il-6〈/em〉) and interleukin 1β (〈em〉il-1β〈/em〉) (〈em〉P〈/em〉 〈 0.05). When KIRA6, the IRE1α kinase inhibitor, was used to block the IRE1α-XBP1s signaling pathway, the Tm and PA-induced pro-inflammatory genes expression was significantly suppressed (〈em〉P〈/em〉 〈 0.05). These data indicated that the IRE1α-XBP1s signaling pathway was involved in the PA-induced inflammatory response in large yellow croaker.〈/p〉〈/div〉 〈/div〉