Rock Bream Research Articles

Amoebic gill disease outbreak in marine fish cultured in Korea.

In 2015, 6.7-60% mortality was observed in black seabream ( Acanthopagrus schlegelii), rock bream ( Oplegnathus fasciatus), and gray mullet ( Mugil cephalus) farmed in the southern coast of Korea. On examination, numerous amoebae were found on the gills of these 3 fish species with detection rate of 100%. Some rock bream and gray mullet were coinfected with bacteria ( Pseudomonas anguilliseptica, Vibrio tapetis, or Vibrio anguillarum). Histologic examination revealed extensive hyperplastic epithelium and lamellar fusion in the gills. Numerous amoebae were seen between gill filaments. The amoebae collected from the 3 fish species had specific 630 bp of a partial 18S rRNA gene fragment for Neoparamoeba perurans. Phylogenetic analysis based on partial 18S rRNA gene nucleotide sequences revealed that these Korean amoeba isolates belonged to the N. perurans group. Based on our results, black seabream, rock bream, and gray mullet were added as new hosts for N. perurans.

A proto-type galectin-2 from rock bream (Oplegnathus fasciatus): Molecular, genomic, and expression analysis, and recognition of microbial pathogens by recombinant protein

A β-galactoside binding lectin, designated as galectin-2, was identified and characterized from rock bream Oplegnathus fasciatus (OfGal-2). The cDNA of OfGal-2 comprised of 692 bp with a coding sequence of 396 bp, encoding a putative polypeptide of 131 amino acids. Gene structure analysis of OfGal-2 revealed a four exon-three intron organization. A single carbohydrate-binding domain containing all seven important residues for carbohydrate binding was located in the third exon, which formed a carbohydrate-binding pocket. Homology screening and sequence analysis demonstrated that OfGal-2 is an evolutionarily conserved proto-type galectin. OfGal-2 transcripts were detected in several healthy fish tissues, with the highest level observed in the intestine, followed by the liver. The expression of OfGal-2 was elevated upon the injection of various mitogenic stimulants and pathogens in a time-dependent manner. Upregulated expression in the liver after tissue injury suggested its role as a damage-associated molecular pattern. Recombinant OfGal-2 protein had hemagglutinating potential and possessed affinity towards lactose and galactose. Moreover, the recombinant protein agglutinated and bound potential pathogenic bacteria and a ciliate. The results of this study indicate that the galectin-2 from rock bream has a potential role in immunity, particularly in the recognition of invading pathogens.

EFFECTS OF DIFFERENT SALINITY LEVELS ON PHYSIOLOGICAL AND HEMATOLOGICAL RESPONSE OF ROCK BREAM Oplegnathus fasciatus

Rock bream Oplegnathus fasciatus is one of economically important marine fish species in East Asia. However, lack of information about the salinity tolerance of rock bream related to its physiological response made this issue were needed to be studied. The present study was conducted to determine the effects of different salinity levels on physiological and hematological response of rock bream in order to obtain its salinity tolerance. Twelve rock breams (total length: 26.9 ± 0.6 cm, body weight: 477.3 ± 61.9 g) were used for experiments. Four experimental groups with three replications were conducted to measure the effects of salinity (5, 15, 25, and 35 practical salinity unit (psu)) on physiological and hematological response of rock bream. Fish were stocked into the chamber inside the closed recirculation system. At the end of each experiment, blood samples were collected. The study revealed that lower salinity exposure had tendency to decrease the physical and chemical properties of blood in rock bream. The value of Na+, Cl-, Ca, Mg, and osmolality showed tendency to decrease with lowering salinity, while cortisol and glucose showed tendency to increase from 35 psu to low salinity environment, indicating the enhancement of fish stress and resulted in fish mortality at 5 psu. The lowest cortisol value was 76.3 ng/mL in 25 psu, and the highest value was 188.8 ng/mL in 5 psu. Meanwhile, the lowest glucose value was 35.3 mg/dL and the highest value was 166.7 mg/dL (P<0.05). Results indicate that rock bream could tolerate lower salinity up to 15 psu.

Identification of a gene encoding a membrane-anchored toll-like receptor 5 (TLR5M) in Oplegnathus fasciatus that responds to flagellin challenge and activates NF-κB

Toll-like receptor 5 (TLR5) recognizes bacterial flagellin and induces the downstream signaling through the myeloid differentiation primary response gene 88 (MyD88) protein to produce proinflammatory cytokines. In this study, we describe a TLR5 membrane form (OfTLR5M) and its adaptor protein MyD88 (OfMyD88) in rock bream, Oplegnathus fasciatus. Both Oftlr5m (6.7 kb) and Ofmyd88 (3.7 kb) genes displayed a quinquepartite structure with five exons and four introns. Protein structure of OfTLR5M revealed the conventional architecture of TLRs featured by an extracellular domain with 22 leucine rich repeats (LRR), a transmembrane domain and an endodomain with TIR motif. Primary OfTLR5M sequence shared a higher homology with teleost TLR5M. The evolutional analysis confirmed that TLR5 identified in the current study is a membrane receptor and the data further suggested the co-evolution of the membrane-anchored and soluble forms of TLR5 in teleosts. Inter-lineage comparison of gene structures in vertebrates indicated that the tlr5m gene has evolved with extensive rearrangement; whereas, the myd88 gene has maintained a stable structure throughout the evolution. Inspection of 5′ flanking region of these genes disclosed the presence of several transcription factor binding sites including NF-κB. Quantitative real-time PCR (qPCR) detected Oftlr5m mRNA in eleven tissues with the highest abundance in liver. In vivo flagellin administration strongly induced the transcripts of both Oftlr5m and Ofmyd88 in gills and head kidney tissues suggesting their ligand-mediated upregulation. In a luciferase assay, HEK293T cells transiently transfected with Oftlr5m and Ofmyd88 demonstrated a higher NF-κB activity than the mock control, and the luciferase activity was intensified when cells were stimulated with flagellin. Collectively, our study represents the genomic, evolutional, expressional and functional insights into a receptor and adaptor molecules of teleost origin that are involved in flagellin sensing.

Improved growth rate and disease resistance in olive flounder, Paralichthys olivaceus, by probiotic Lactococcus lactis WFLU12 isolated from wild marine fish

The use of probiotics is a strategy employed to improve host health status and to prevent infectious diseases. The current study was aimed at investigating the diversity of lactic acid bacteria (LAB) and Bacillus species in the gastrointestinal tracts of wild marine fishes, as well as the beneficial effects of Lactococcus lactis WFLU12 as a host-derived probiotics in olive flounder. In marine fishes, wild olive flounder and rock bream were shown to be good sources of LAB and Bacillus isolation, respectively. Some isolates, including the strain WFLU12, have shown stronger inhibitory activity against various aquatic bacterial pathogens and more tolerance to low pH and bile acids compared to some strains isolated from sources other than marine. Lc. lactis WFLU12 was found to confer to olive flounder protection against streptococcosis caused by Streptococcus parauberis through competitive exclusion and increased innate immune responses. Interestingly, the natural infection rate in the probiotic fed group (33%=10/30) was significantly lower than that in the control group (60%=18/30). None of the nisin Z and colicin V-producing probiotic-fed fish were naturally infected by S. parauberis during the feeding period. In addition, more importantly, this promising probiotic strain significantly promoted fish growth along with better feed conversion and specific growth rate. This study demonstrates that the use of host-derived probiotics can offer a significant advantage in terms of optimum survival and function in the gastrointestinal tract of the intended host. Statement of relevanceIn this study, host-derived probiotic strain outperforms elimination of pathogen through competitive exclusion in the gastrointestinal tract and increased innate immune responses. More importantly, this promising probiotic strain significantly promoted fish growth along with better feed conversion. This study will provide insight into how optimal probiotics should be selected and developed. It might facilitate the replacement of commercial fish probiotic products originated from terrestrial sources with host-derived probiotics in the near future.

Evaluation of the Dietary Effect of Hallabong Peel Oil on Growth, Hematological, and Immune Gene Expression in Rock Bream, Oplegnathus fasciatus Challenged with Edwardsiella tarda

In the present study we evaluated the dietary effect of Hallabong peel oil (HPO) on growth, disease resistance, and immune gene expression of rock bream, Oplegnathus fasciatus challenged with Edwardsiella tarda after a 4 week feeding trial with 5 treatments: control-C, probiotic–P, HPO (0.1%), HPO (0.5%), and P+HPO, diets. All fish groups were assessed for growth performance, innate immune parameters, serum biochemical profile, and immune gene expression in head kidney on 2nd, and 4th week, and 1st, 3rd and 7th day post infection with Edwardsiella tarda. Fish fed the HPO enriched diets showed increased growth performance with significantly decreased (P&gt;0.05) mortality compared with the control and probiotic diet groups. The positive effects of HPO enriched diet were also found in all assessed innate immune and biochemical parameters which included increased respiratory burst and lysozyme activity, with significantly increased erythrocyte and leukocytes counts, increased serum protein, decreased glucose, triglycerides, cholesterol level in serum compared with control diet fed fish. Moreover, the probiotic bacterial count in the intestine of fish was enhanced with the HPO diet and the P+HPO diet compared to fish fed the probiotic diet. The head kidney of HPO enriched diet fed fish showed up-regulated expression of inflammatory cytokines genes such as TNFα, IL-1β, and FST, after 4th week of feeding trial which was increased ~2 to 3 times on 1dpi and 3 dpi. These results indicate that limonene rich (91.26%), HPO enriched diets enhance growth and immunity and enhance disease resistance of Oplegnathus fasciatus challenged against E. tarda.

Dietary Effect of Probiotic Bacteria, Bacillus amyloliquefaciens-JFP2 on Growth and Innate Immune Response in Rock Bream Oplegnathus fasciatus, Challenged with Streptococcus iniae

The present study was conducted to evaluate the use of Bacillus amyloliquefaciens as a probiotic source in the diet of rock bream, Oplegnathus fasciatus. A total of 210 fingerlings (25.4 ± 0.13 g) were divided into two groups, three replicates each; control (C) and probiotic (P) group (35×3=105 fish per group). C group was fed a basal diet without probiotic while P group was fed with B. amyloliquefaciens spores at a concentration of 1.4 ×106 colony-forming units per gram (CFU/g) of feed. After 90 days, P group fish showed significant improvements in body weight (BW), weight gain (WG), specific growth rate (SGR) and food conversion ratio (FCR) when compared with C group fish. Also, in bi-weekly assessments, serum protein and glucose level showed a significant increase (p&lt;0.05) in probiotic fed fish compared with the control group. Similar increase in serum antioxidant and lysozyme activity was found in the probiotic fed fish group. However, there was a significant decrease in triglyceride and total cholesterol along with increased alanine aminotransferase (ALT) and aspartate amonotransferase (AST) levels in probiotic diet fed fish compared to the control. After seventy days the fish were challenged with Streptococcus iniae and mortality of the probiotic fed fish was significantly lower (p&lt;0.05) when compared with the control. These results demonstrate the benefit of incorporation of B. amyloliquefaciens as a feed supplement to improve the health status of Oplegnathus fasciatus challenged with Streptococcus iniae.

Dietary Effect of Probiotic Bacteria, Bacillus amyloliquefaciens-JFP2 on Growth and Innate Immune Response in Rock Bream Oplegnathus fasciatus, Challenged with Streptococcus iniae

The present study was conducted to evaluate the use of Bacillus amyloliquefaciens as a probiotic source in the diet of rock bream, Oplegnathus fasciatus. A total of 210 fingerlings (25.4 ± 0.13 g) were divided into two groups, three replicates each; control (C) and probiotic (P) group (35×3=105 fish per group). C group was fed a basal diet without probiotic while P group was fed with B. amyloliquefaciens spores at a concentration of 1.4 ×106 colony-forming units per gram (CFU/g) of feed. After 90 days, P group fish showed significant improvements in body weight (BW), weight gain (WG), specific growth rate (SGR) and food conversion ratio (FCR) when compared with C group fish. Also, in bi-weekly assessments, serum protein and glucose level showed a significant increase (p&lt;0.05) in probiotic fed fish compared with the control group. Similar increase in serum antioxidant and lysozyme activity was found in the probiotic fed fish group. However, there was a significant decrease in triglyceride and total cholesterol along with increased alanine aminotransferase (ALT) and aspartate amonotransferase (AST) levels in probiotic diet fed fish compared to the control. After seventy days the fish were challenged with Streptococcus iniae and mortality of the probiotic fed fish was significantly lower (p&lt;0.05) when compared with the control. These results demonstrate the benefit of incorporation of B. amyloliquefaciens as a feed supplement to improve the health status of Oplegnathus fasciatus challenged with Streptococcus iniae.

Evaluation of the Dietary Effect of Hallabong Peel Oil on Growth, Hematological, and Immune Gene Expression in Rock Bream, Oplegnathus fasciatus Challenged with Edwardsiella tarda

In the present study we evaluated the dietary effect of Hallabong peel oil (HPO) on growth, disease resistance, and immune gene expression of rock bream, Oplegnathus fasciatus challenged with Edwardsiella tarda after a 4 week feeding trial with 5 treatments: control-C, probiotic–P, HPO (0.1%), HPO (0.5%), and P+HPO, diets. All fish groups were assessed for growth performance, innate immune parameters, serum biochemical profile, and immune gene expression in head kidney on 2nd, and 4th week, and 1st, 3rd and 7th day post infection with Edwardsiella tarda. Fish fed the HPO enriched diets showed increased growth performance with significantly decreased (P&gt;0.05) mortality compared with the control and probiotic diet groups. The positive effects of HPO enriched diet were also found in all assessed innate immune and biochemical parameters which included increased respiratory burst and lysozyme activity, with significantly increased erythrocyte and leukocytes counts, increased serum protein, decreased glucose, triglycerides, cholesterol level in serum compared with control diet fed fish. Moreover, the probiotic bacterial count in the intestine of fish was enhanced with the HPO diet and the P+HPO diet compared to fish fed the probiotic diet. The head kidney of HPO enriched diet fed fish showed up-regulated expression of inflammatory cytokines genes such as TNFα, IL-1β, and FST, after 4th week of feeding trial which was increased ~2 to 3 times on 1dpi and 3 dpi. These results indicate that limonene rich (91.26%), HPO enriched diets enhance growth and immunity and enhance disease resistance of Oplegnathus fasciatus challenged against E. tarda.

Development of PCR method for detecting Kudoa iwatai (Myxozoa: Multivalvulida) from rock bream Oplegnathus fasciatus.

We developed a PCR assay targeting the 28S rDNA of Kudoa iwatai (Multivalvulida: Myxozoa) and investigated the prevalence of infection in rock bream Oplegnathus fasciatus, which is commercially an important aquaculture species in Korea, with this assay. Detection limit of the PCR assay was 2.5fg/μl with plasmid DNA and 8.6×103 spores/ml with purified spores, respectively. This PCR assay did not amplify DNA of other Kudoa species (Kudoa septempunctata, Kudoa lateolabracis, Kudoa thyrsites) tested. Sliced muscles of whole body from 318 rock bream (wild and cultured) were examined by this PCR assay and also with the naked eyes. All of the wild fish did not produce amplicons nor did harbor visible Kudoa cysts (0/70). Three of the cultured fish were PCR-positive and also harbored visible Kudoa cysts (3/248, 1.2%). The sequences of amplicons (574bp) were 100% identical with those of the K. iwatai already registered in Genbank. When the visceral organs of these three fish were examined, visible cysts were not found, but one stomach sample was found to be PCR-positive. There was no difference in the prevalence of infection estimated by PCR assay and the presence of visible Kudoa cysts in our samples. This is thought to be because the development of K. iwatai is already completed and only mature Kudoa cysts existed in our samples.

Expression Analysis of Lily Type Lectin Isotypes in the Rock Bream, Oplegnathus fasciatus: in the Tissue, Developmental Stage and Viral Infection.

ABSTRACTLectins belong to the pattern-recognition receptors (PRRs) class and play important roles in the recognition and elimination of pathogens via the innate immune system. Recently, it was reported that lily-type lectin-1 is involved when a pathogen attacks in the early immune response of fish. However, this study is limited to information that the lectin is involved in the innate immune response against viral infection. In the present study, the lily-type lectin-2 and -3 of Oplegnathus fasciatus (OfLTL-2 and 3) have been presented to be included B-lectin domain and two D-mannose binding sites in the amino acid sequence that an important feature for the fundamental structure. To investigate the functional properties of OfLTLs, the tissue distribution in the healthy rock bream and temporal expression during early developmental stage analysis are performed using quantitative real-time PCR. OfLTL-2 and 3 are predominantly expressed in the liver and skin, but rarely expressed in other organ. Also, the transcripts of OfLTLs are not expressed during the early developmental stage but its transcripts are increased after immune-related organs which are fully formed. In the challenge experiment with RBIV (rock bream iridovirus), the expression of OfLTLs was increased much more strongly in the late response than the early, unlike previously known. These results suggest that OfLTLs are specifically expressed in the immune-related tissues when those organs are fully formed and it can be inferred that the more intensively involved in the second half to the virus infection.

Establishment of rock bream Oplegnathus fasciatus embryo (RoBE-4) cells with cytolytic infection of red seabream iridovirus (RSIV)

Red seabream iridovirus (RSIV) is a member of genus Megalocytivirus in the family Iridoviridae. RSIV infection causes significant economic losses of marine-fishes in East Asian countries. Grunt fin (GF) cell line has been commonly used for culturing RSIV. However, it is not suitable for definite evaluation of infectivity titer of RSIV because cells infected with RSIV are not completely cytolysed. Thus, we established a new cell line, RoBE-4, from rock bream (Oplegnathus fasciatus) eyed-egg embryos in this study. Morphologically, RoBE-4 cells were fibroblastic-like. They have been stably grown over two-years with 60 passages using Leibovitz’s L-15 medium containing 10% (v/v) fetal bovine serum. RoBE-4 cells infected with RSIV exhibited cytopathic effects (CPE) with cell rounding. They were cytolysed completely after ≥2 weeks of culture. Numerous RSIV particles with icosahedral morphology of approximately 122nm in diameter were observed in cytoplasmic area of infected RoBE-4 cells. The RSIV-suceptibility and amount of extracellular RSIV released by RoBE-4 cells were 100-fold higher than those by GF cells. RSIV cultured with RoBE-4 cells was highly virulent to rock bream in infection experiments. Therefore, using RoBE-4 cells instead of GF cells will enable accurate and sensitive measurement of RSIV infectivity. In addition, RoBE-4 cells might be used to produce RSIV vaccine in the future with significant reduction in cost.

Production and characterization of monoclonal antibodies against recombinant ORF 049L of rock bream iridovirus

Abstract Iridovirus has been detected in more than 20 fish species, caused severe systemic diseases. Rock bream iridovirus (RBIV) is a known causative agent of epizootics in cultured Oplegnathus fasciatus in China. In this study, monoclonal antibodies (mAbs) against the recombinant RBIV protein ORF 049L were employed to further elucidate the pathogenesis of the virus. Briefly, mAbs against recombinant ORF 049L were prepared. Western blot analysis was performed to assess the ability of the mAbs to bind to the recombinant protein, and the sensitivity of mAbs to RBIV-infected rock bream was analyzed by dot blot analysis, an indirect immunofluorescence assay, and one-step PCR analysis. Eight mAbs against recombinant ORF 049L were developed, of which three (3F8, 2H11, and 2B7) were sensitive to RBIV-infected rock bream. These mAbs provide useful tools to analyze the location and replication in vivo of structural and functional proteins, and the pathogenesis of RBIV in aquatic organisms.

Cloning and expression analysis of innate immune genes from red sea bream to assess different susceptibility to megalocytivirus infection.

As suggested by the Office International des Epizooties (OIE), fishes belonging to the genus Oplegnathus are more sensitive to megalocytivirus infection than other fish species including red sea bream (Pagrus major). To assess the roles of the innate immune response to these different susceptibilities, we cloned the genes encoding inflammatory factors including IL-8 and COX-2, and the antiviral factor like Mx from red sea bream for the first time and performed phylogenetic and structural analysis. Analysed expression levels of IL-1β, IL-8 and COX-2 and the antiviral factor like Mx genes performed with invivo challenge experiment showed no difference in inflammatory gene expression or respiratory burst activity between red sea bream androck bream (Oplegnathus fasciatus). However, the Mx gene expression levels in red sea bream weremarkedly higher than those in rock bream, suggesting the importance of type I interferon (IFN)-induced proteins, particularly Mx, during megalocytivirus infection, rather than inflammation-related genes. The invitro challenge experiments using embryonic primary cultures derived from both fish species showed no difference in cytopathic effects (CPE), viral replication profiles, and inflammatory and Mx gene expression pattern between the two fish species.

The optimum dietary docosahexaenoic acid level based on growth and non-specific immune responses in juvenile rock bream,Oplegnathus fasciatus

A feeding trial was conducted to determine the optimum level and effect of incremental dietary levels of docosahexaenoic acid (DHA, 22:6n-3) on growth and non-specific immune responses in juvenile rock bream, Oplegnathus fasciatus. A basal diet without DHA supplementation was used as a control, and six other diets were prepared by supplementing with 4, 8, 12, 16, 20 or 40 g kg−1 DHA. These diets included no eicosapentaenoic acid and/or arachidonic acid contents. The actual DHA concentrations of the diets were 1, 4.8, 8.9, 13.1, 17.6, 21.2 and 41.4 g kg−1 diet (DHA1.0, DHA4.8, DHA8.9, DHA13.1, DHA17.6, DHA21.2 and DHA41.4 respectively). At the end of feeding trial, final body weight, weight gain, specific growth rate and feed efficiency of fish fed the DHA13.1, DHA17.6, DHA21.2 and DHA41.4 diets were significantly higher than those fed the other diets (P < 0.05). The broken-line analysis of weight gain indicates that the optimum dietary DHA level is 11.9 g kg−1. Fish fed DHA1.0 had the highest hepatosomatic index, an increase in plasma cholesterol, triglyceride, low-density lipoprotein and aspartate aminotransferase levels, as well as a decrease in high-density lipoprotein. Superoxide dismutase activity of fish fed DHA13.1 and DHA17.6 diets was significantly higher than that of fish fed DHA1.0, DHA4.8 and DHA8.9 diets. Fish fed the DHA17.6, DHA21.2 and DHA41.4 diets showed significantly higher lysozyme activity than those fish fed DHA1.0, DHA4.8 and DHA8.9 diets. Therefore, the optimum dietary DHA level could be greater than 11.9 g kg−1 but less than 13.1 g kg−1 in diet.

Molecular cloning and expression analysis of a new lily-type lectin in the rock bream, Oplegnathus fasciatus

A new lily-type lectin RbLTL was identified from rock bream (Oplegnathus fasciatus) and its expression analysed. In this study, a new lily-type lectin gene (RbLTL) was cloned from rock bream using expressed sequence tag (EST) analysis. The full-length RbLTL cDNA was encoding a 117-amino acid protein. The deduced amino acid sequence of RbLTL contained all of the conserved features crucial for its fundamental structure, including B-lectin domain and three d-mannose binding sites. RbLTL mRNA was predominately expressed in the gills, with reduced expression noted in intestine tissue. Expression analysis of time series sampled fertilized eggs revealed that expression gradually increased 1, 3, 12, and 24 h: However, expression decreased at 36 h. RbLTL expression was differentially up-regulated in rock bream gills challenged with Streptococcus iniae, Edwardsiella tarda and RSIV. Our results revealed that novel rock bream lily-type lectin may be an important molecule involved in pattern recognition and pathogen elimination in the innate immunity of rock bream.

Complement factor D homolog involved in the alternative complement pathway of rock bream (Oplegnathus fasciatus): Molecular and functional characterization and immune responsive mRNA expression analysis.

The complement system serves conventional role in the innate defense against common invading pathogens. Complement factor D (CfD) is vital to alternative complement pathway activation in cleaving complement factor B. This catalytic reaction forms the alternative C3 convertase that is crucial for complement-mediated pathogenesis. In this study, rock bream (Oplegnathus fasciatus) CfD (OfCfD) was characterized and OfCfD mRNA expression was investigated. OfCfD encodes 277amino acids (aa) for a 30-kDa polypeptide. A domain analysis of the deduced OfCfD aa sequence showed a single serine protease trypsin superfamily domain, a serine active region, three active sites, and three substrate-binding sites. Pairwise sequence comparisons indicated that OfCfD has the highest identity (84.5%) with Oreochromis niloticus CfD. The phylogenetic tree revealed a common ancestral origin of CfD members, with fish CfD distinct from other vertebrate orthologs. The structural arrangement of the OfCfD gene (2451bp) contained five exons interrupted by four introns. A spatial transcriptional analysis indicated that OfCfD transcripts constitutively expressed in all of the examined rock bream tissues, and that they were highest in the spleen and liver. In addition, OfCfD transcripts were immunologically upregulated by lipopolysaccharide (LPS) (12h p.i.), Streptococcus iniae (12h p.i.), rock bream iridovirus (RBIV) (6-12h p.i.), and poly I:C (6h p.i.) in spleen tissue. OfCfD is a trypsin protease and its recombinant protein showed strong protease activity similar to that of trypsin, indicating its catalytic function in the alternative pathway. Together, our findings suggest that OfCfD might be involved in immune responses in rock bream.

Cooperation of erythrocytes with leukocytes in immune response of a teleost Oplegnathus fasciatus

The most abundant cell type in the blood of mammals and fish is erythrocyte. Unlike mammalian erythrocyte, fish erythrocyte is nucleated. The functional differentiation of teleost erythrocyte is insufficient compared with that of mammals. Therefore, fish erythrocyte may have different functions from that of mammals. Functional interaction between erythrocyte and leukocyte was confirmed by the cDNA microarray newly constructed in this study to investigate characterization of rock bream erythrocyte. In this study, different immune related genes of erythrocytes were annotated by microarray analysis. Microarray analysis showed that a total of 338 genes were up-regulated in co-cultured erythrocytes with leukocytes by LPS stimulation when comparing to erythrocytes stimulated LPS. Many genes in erythrocyte of rock bream were up-regulated in presence of leukocytes, suggesting that erythrocytes interact with leukocytes to trigger expression of various genes associated with the immune system. Our results provide valuable information that direct and indirect immunological function of fish erythrocyte.

Analysis of immune gene expression in rock bream spleen after rock bream iridovirus infection by transcriptome sequencing

The results of amputation free survival (AFS) of a cohort of 887 caucasian patients is shown.The data is based on further analyses of data presented in Jalkanen et al. (2016) [1]. The 36-month amputation free survival of patients divided in new crural vessel disease classification (Crural Index), aortoiliac TASC II classification, femoropopliteal TASC II classification and most severe segment is presented. Also, in depth demographic data is presented for each Crural Index group Jalkanen et al., 2016 [1].

Effects of various LED light spectra on antioxidant and immune response in juvenile rock bream, Oplegnathus fasciatus exposed to bisphenol A.

Bisphenol A (BPA) is a monomer used in plastics and plasticizers. As an environmental toxin included in industrial wastewater, it contaminates the aquatic environment and is known to cause endocrine disruption in fish. Particular wavelengths of light-emitting diodes (LEDs) are known to affect the endocrine regulation of fish. The present study aimed to investigate the effects of green and red LED light on the antioxidant and immune systems in juvenile rock bream (Oplegnathus fasciatus) exposed to BPA. We used green and red LED exposure at two intensities (0.3 and 0.5W/m(2)) for 1, 3, and 5 days. We measured liver mRNA expression and plasma levels of antioxidant enzyme superoxide dismutase (SOD) and caspase-3. Furthermore, we measured plasma levels of hydrogen peroxide (H2O2), lipid peroxidation (LPO), melatonin, and immunoglobulin M (IgM). DNA damage and apoptotic activity were measured using comet and terminal transferase dUTP nick end labeling (TUNEL) assays, respectively. We found that SOD, H2O2, and LPO increased significantly, whereas melatonin and IgM decreased significantly, suggesting that BPA induces oxidative stress and reduces immune function. Likewise, both DNA damage and apoptotic activity increased following BPA exposure. However, we found that exposure to green LED light effectively reduced the detrimental effects induced by BPA, including decreasing DNA damage, apoptotic activity, SOD mRNA expression, and plasma levels of SOD, H2O2, and LPO. Likewise, the plasma levels of melatonin and IgM increased. Thus, our results indicate that green light conditions effectively reduces oxidative stress and promotes the immune function in juvenile rock bream.