Rock Bream Research Articles

Improving effectiveness of environmental enrichment: the role of light intensity in rock bream (Oplegnathus fasciatus) rearing

Environmental enrichment is a potential approach to enhance the fitness and survival of hatchery fish stocked for conservation purposes. Its effectiveness presumably depends not only on ecologically relevant features of physical enrichment but also on light condition applied. Despite the importance, few studies have reported on the role of light in enriched rearing. This study sought to optimize the enriched rearing of juvenile rock bream (Oplegnathus fasciatus) particularly regarding their physiological and behavioral performance by regulating light intensity. Juveniles were reared in either standard tanks (no enrichment) or enriched tanks with polyethylene ropes simulating drifting algae at three different light intensities (400, 800 or 1600 lux) for 32 days. Our findings revealed that cortisol levels of the fish increased with light intensity even in the presence of the physical structure. No significant effect of light intensity was detected on growth. Compared to standard fish, fish reared with enrichment showed higher levels of caudal fin damage and cortisol at all the light intensities, suggesting increased chronic stress; in addition, the fish displayed higher shelter-seeking behavior. These results highlight the importance of maintaining low light intensities to minimize chronic stress in the fish. Moreover, adjustments to the enriched rearing condition are recommended to mitigate adverse effects on physiological aspects before incorporated into practical implementation in stock enhancement programs.

Characterization of IgD and IgT with their expressional analysis following subtype II megalocytivirus vaccination and infection in rock bream (Oplegnathus fasciatus)

In this study, heavy chain genes of IgD and IgT were sequenced and characterized their gene expression in rock bream (Oplegnathus fasciatus). Rock bream (RB)-IgD cDNA is 3319 bp in length and encodes a leader region, variable domains, a μ1 domain, and seven constant domains (CH1–CH7). A membrane-bound (mIgT) and secretory form (sIgT) of RB-IgT cDNAs are 1902 bp and 1689 bp in length, respectively, and encode a leader region, variable domains, four constant domains (CH1–CH4) and C-terminus. Their predicted 3D-structure and phylogenetic relation were similar to those of other teleost. In healthy fish, RB-IgD and mIgT gene expressions were higher in major lymphoid organs and blood, while RB-sIgT gene was more highly expressed in midgut. IgT expressing cells were detected in melano-macrophage centers (MMC) of head kidney in immunohistochemistry analysis. Under immune stimulation in vitro, RB-IgD and IgT gene expressions were upregulated in head kidney and spleen cells by bovine serum albumin or a rock bream iridovirus (RBIV) vaccine. In vivo, their expressions were significantly upregulated in head kidney, blood, and gill upon vaccination. Especially, RB-mIgT gene expression in head kidney and blood was upregulated at day 3 after vaccination while upregulated at earlier time point of day 1 by challenge with RBIV. This may suggest that memory cells might be produced during the primary response by vaccination and rapidly proliferated by secondary immune response by viral infection. RB-sIgT gene expression was highly upregulated in peripheral blood in vaccinated fish after viral infection, indicating that IgT plays an important role in systemic immune response as well as mucosal immune system. Our findings provide information on the role of RB-IgT in adaptive immunity during vaccination and viral infection in the vaccinated fish.

Effect of rock bream iridovirus (RBIV) contained tissue intake on rock bream (Oplegnathus fasciatus) mortality and blood cell distribution

Rock bream (Oplegnathus fasciatus) is one of the highly priced cultured marine fish in Korea. Rock bream iridovirus (RBIV) outbreaks in aquaculture farms may involve environmental factors, co-infection with other pathogenic microorganisms and grounded (raw) fish feed. This study evaluated the effects of RBIV-containing tissue intake on mortality and oral transmission in rock bream. Virus-containing tissues administered to rock bream [50 mg (1.53 × 108/major capsid protein, MCP gene copies) to 2400 mg (7.34 × 109)] held at 23 °C lead to 100 % mortality by 27 days post administration. Interestingly, the mortality rates were not viral dose- or concentration dependent. Further, high MCP gene copy numbers were observed in the gill, liver, intestine, stomach, spleen, heart, kidney, brain and muscle tissues (viral load range of 3.03 × 106 to 4.01 × 107/mg, average viral load 1.70 × 107/mg) of dead rock bream. Moreover, a high viral load was detected in the intestine and stomach, where the virus was directly administered. This indicated that the intake of RBIV-containing tissue feed weakens the intestinal mucosal immunity and increases viral load in the intestine. Moreover, the levels of complete blood cell count (CBC) indicators, such as red blood cell (RBC), hemoglobin (HGB) and hematocrit (HCT) significantly decreased from 15 dpi with red blood cell distribution width (RDW), and white blood cells (lymphocyte, monocyte and granulocyte) significantly increased from the initial to later stage of infection. These results highlight the significance of blood-mediated indicators against RBIV infection in rock bream. We demonstrate the existence of an oral transmission route for RBIV in rock bream. Our findings indicate that pathogen-containing feed is an important risk factor for disease outbreaks in rock bream.

Evaluation of Formalin-Inactivated Vaccine Efficacy against Red Seabream Iridovirus (RSIV) in Laboratory and Field Conditions.

Red seabream iridovirus (RSIV) is a major cause of marine fish mortality in Korea, with no effective vaccine available since its first occurrence in the 1990s. This study evaluated the efficacy of a formalin-killed vaccine against RSIV in rock bream under laboratory and field conditions. For the field trial, a total of 103,200 rock bream from two commercial marine cage-cultured farms in Southern Korea were vaccinated. Farm A vaccinated 31,100 fish in July 2020 and monitored them for 18 weeks, while farm B vaccinated 30,700 fish in August 2020 and monitored them for 12 weeks. At farm A, where there was no RSIV infection, the vaccine efficacy was assessed in the lab, showing a relative percentage of survival (RPS) ranging from 40% to 80%. At farm B, where natural RSIV infections occurred, cumulative mortality rates were 36.43% in the vaccinated group and 80.32% in the control group, resulting in an RPS of 54.67%. The RSIV-infectious status and neutralizing antibody titers in serum mirrored the cumulative mortality results. This study demonstrates that the formalin-killed vaccine effectively prevents RSIV in cage-cultured rock bream under both laboratory and field conditions.

Infection dynamics and shedding kinetics of RSIV in red sea bream (Pagrus major) using immersion and cohabitation challenge models

Red sea bream iridovirus (RSIV) poses a significant threat to global aquaculture and wild fish populations, leading to substantial economic and ecological losses. Despite its recognition as a major viral disease, our understanding of the dynamics of RSIV infection and transmission in naturalistic conditions remains limited. Therefore, the present study aimed to investigate the kinetics, shedding, and interspecies transmission of RSIV in red sea bream (Pagrus major) through immersion and cohabitation challenge models. The primary objectives were to (1) elucidate the pathogenicity of RSIV in red sea bream through naturalistic immersion challenges, (2) analyze the viral load and shedding dynamics in RSIV-infected fish, and (3) assess the risk of horizontal transmission to other species. Our results indicated that red sea bream exposed to high doses of RSIV (101–107 RSIV copies/mL) experienced rapid cumulative mortality, particularly at 25 °C. Notably, red sea bream infected by immersion at 25 and 15 °C showed a significant increase in RSIV shedding into seawater when the viral load exceeded approximately 102.48 RSIV copies/mg, highlighting a critical threshold for understanding disease spread in aquaculture settings. Histopathological analysis revealed that the spleen and kidney were primarily affected, with significant lesion scores correlating with viral loads. We observed a strong correlation between viral load and histopathological infection grades, particularly in these primary tissues. Furthermore, cohabitation experiments highlighted the risk of RSIV transmission to other fish species, including rock bream (Oplegnathus fasciatus) and flathead grey mullet (Mugil cephalus), indicating the possibility of horizontal spread in multi-species farming systems. These findings emphasize the importance of continuous surveillance and robust biosecurity measures in aquaculture systems to manage RSIV infection effectively. Overall, this study contributes to understanding RSIV dynamics in red sea bream, offering insights into the impact of this virus in aquaculture environments, particularly concerning viral shedding and transmission.

Molecular characterization, antibacterial and immunoregulatory activities of liver-expressed antimicrobial peptide 2 in black rockfish, Sebastes schlegelii

LEAP2 (liver expression antimicrobial peptide 2), is an antimicrobial peptide widely found in vertebrates and mainly expressed in liver. LEAP2 plays a vital role in host innate immunity. In teleosts, a number of LEAP2 homologs have been reported, but their in vivo effects on host defense are still limited. In this study, a LEAP2 homolog (SsLEAP2) was identified from black rockfish, Sebastes schlegelii, and its structure, expression as well as biological functions were analyzed. The results showed that the open reading frame of SsLEAP2 is 300 bp, with a 5′- untranslated region (UTR) of 375 bp and a 3’ - UTR of 238 bp. The deduced amino acid sequence of SsLEAP2 shares the highest overall identity (96.97%) with LEAP2 of Sebastes umbrosus. SsLEAP2 possesses conserved LEAP2 features, including a signal peptide sequence, a prodomain and a mature peptide, in which four well-conserved cysteines formed two intrachain disulphide domain. The expression of SsLEAP2 was highest in liver and could be induced by experimental infection with Listonella anguillarum, Edwardsiealla piscicida and Rock bream iridovirus C1 (RBIV-C1). Recombinant SsLEAP2 (rSsLEAP2) purified from Escherichia coli was able to bind with various Gram-positive and Gram-negative bacteria. Further analysis showed that rSsLEAP2 could enhance the respiratory burst activity, and induce the expression of immune genes including interleukin 1-β (IL-1β) and serum amyloid A (SAA) in macrophages; additionally, rSsLEAP2 could also promote the proliferation and chemotactic of peripheral blood lymphocytes (PBLs). In vivo experiments indicated that overexpression of SsLEAP2 could inhibit bacterial infection, and increase the expression level of immune genes including IL-1β, tumor necrosis factor ligand superfamily member 13B (TNF13B) and haptoglobin (HP); conversely, knock down of SsLEAP2 promoted bacterial infection and decreased the expression level of above genes. Taken together, these results suggest that SsLEAP2 is a novel LEAP2 homolog that possesses apparent antibacterial activity and immunoregulatory property, thus plays a critical role in host defense against pathogens invasion.

Comparison of Genetic Diversity between Hatchery‐Reared and Wild Rock Bream (Oplegnathus fasciatus) Based on Microsatellite Markers and Mitochondrial COI Sequences

Hatchery release of rock bream (Oplegnathus fasciatus) is common in the Zhoushan Sea region of the East China Sea as an efficient method for stock enhancement. Clarifying the genetic effects of farm‐raised populations on wild ones is important for understanding the health of fish stocks. Thus, this study collected six rock bream stocks (two wild, three hatchery‐reared, and one parental) for genetic diversity analysis using microsatellite markers and mitochondrial sequences. Although the results of mitochondrial sequence analysis showed that 61.26% of the genetic diversity resided among groups, indicating a high degree of genetic differentiation among wild, hatchery‐reared, and parental groups, the results of microsatellite analysis showed that 30% of the genetic diversity resided within the populations, indicating no obvious genetic differentiation among different groups. Our study suggests that though releasing hatchery‐reared rock bream for stock enhancement may not noticeably decrease the genetic diversity of wild populations in the Zhoushan Sea in the short term, long‐term genetic evaluations shall be taken to monitor genetic diversity in wild rock bream and ensure that the populations remain healthy.

Targeting the Major Capsid Protein of Rock Bream Iridovirus by Trapa Natans Phytochemical Compounds; a Molecular Docking Approach

Asian rock bream aquaculture is challenged by iridovirus, a pathogen with no effective vaccines or antiviral agents. The present study aimed to identify natural antiviral compounds from Trapa natans, an aquatic plant, with potential to target the activities of major capsid protein in rock bream iridovirus (RBIVmcp). The investigation commenced by constructing a novel 3D homology structure for RBIVmcp using I-TASSER server, followed by refinement and validation through galaxy refiner, ModRefiner, PDBSum, ProSA, and ERRAT servers. Active binding regions of refined protein structures were employed for molecular docking simulations using PyRx and AutoDock Vina tools. Leveraging IMPPAT and PubChem servers, a total of 13 phytochemical compounds from Trapa natans were identified in silico and subjected to docking analysis against RBIVmcp. Among them, Ellagic acid, Galactoarabinan, and Thiamine exhibited the most favorable docking scores of -5.1 kcal/mol, -6.4 kcal/mol, and -5.9 kcal/mol, respectively. Assessment of compounds' toxicity and ADME properties by Swiss-ADME and ProTox-II servers indicated their non-hepatotoxic, non-carcinogenic, non-immunotoxic, and non-mutagenic nature within environments. These findings suggest aforementioned compounds might act as promising antiviral agents against RBIV by interacting with its major capsid protein. However, in vitro and in vivo studies are essential to validate their efficacy.

Environmental enrichment improves behaviors rather than the growth and physiology of rock bream Oplegnathus fasciatus.

Environmental enrichment has the potential to improve the welfare and post-release survival of hatchery fish stocked for conservation purposes. However, the effectiveness of environmental enrichment is partly dependent on the fish species, life stage, and specific enrichment structure used. To enhance the effectiveness of environmental enrichment, it is crucial to focus on characteristic differences in enrichment structures, such as type and level. This study investigated how differences in enrichment type and level affected physiological and behavioral aspects of the welfare of pre-release juvenile rock bream Oplegnathus fasciatus by evaluating growth performance, basal and stressed cortisol levels, antioxidant enzyme activities, and exploratory behaviors regarding anxiety and flexibility. Fish were reared for 4 weeks in different enrichment treatments: barren, low-level cover structure, high-level cover structure, low-level interference structure (LI), and high-level interference structure (HI). The results revealed that fish reared with the LI treatment showed less anxiety and greater flexibility with respect to exploratory behaviors, without oxidative damage being detected. Despite exhibiting less anxiety as well, fish reared in the HI treatment had oxidative damage, indicated by lower superoxide dismutase activity, compared to those in the barren treatment. In addition, none of these enrichment structures enhanced growth performance or mitigate chronic and acute stress responses. Overall, the low-level interference structure may be more favorable in promoting the behavioral welfare of the fish. Application of this type and level of enrichment may increase the survival of the hatchery fish after release, which is critical to stocking success.

Abnormalities in red blood cell production and pathogenesis of anemia in the progression of rock bream iridovirus (RBIV)

Rock bream iridovirus (RBIV), belonging to Megalocytivirus, causes severe mortality in rock bream. Almost all deaths associated with RBIV are accompanied by splenic enlargement and anemia. Although red blood cells (RBCs) are involved in the immune response against viral infections, their involvement in rock bream has not yet been studied in terms of the immune response against RBIV. In this study, the viral replication patterns, blood characteristics and anemia-related factors were evaluated in rock bream post RBIV infection. The virus-infected RBCs of rock bream demonstrated similarities in the expression levels of hemoglobins (HGB) (α and β), cytokine-dependent hematopoietic cell linker (CLNK) and hematopoietic transcription factor GATA (GATA), with significantly decreasing levels from 4 days post infection (dpi) to 17 (dpi), when the viral replication was at its peak. This suggests that the expression of blood-related genes is inadequate for HGB synthesis and RBC production, thereby causing anemia leading to death. Moreover, the levels of complete blood cell count (CBC) indicators, such as RBCs, HGB and hematocrit (HCT), significantly decreased from 10 to 17 dpi. This phenomenon suggests that blood-related gene expression and/or RBC-, HGB- and HCT-related levels are critical factors in RBIV-induced anemia and disease progression. These results highlight the significance of blood-mediated immune responses against RBIV infection in rock bream. Understanding blood-related gene levels to identify blood-related immune response interactions in rock bream will be useful for development of future strategies in controlling RBIV diseases in rock bream.

Red sea bream iridovirus infection downregulates inflammation-related genes in the spleen of rock bream (Oplegnathus fasciatus).

This study investigated the kinetics of red sea bream iridovirus and host gene expression during infection in rock bream (Oplegnathus fasciatus), a species highly sensitive to the virus. After intraperitoneal injection of the viral solution at 104 TCID50/fish, the viral genome copy number in the spleen was 104.7 ± 0.2 and 105.9 ± 0.4 copies/μg DNA at 3 and 5 days post-injection (dpi), respectively. Using transcriptomic analyses via MiSeq, viral gene transcripts were detected at 3 and 5 dpi. Six genes including RING-finger domain-containing protein and laminin-type epidermal growth factor-like domain genes were significantly expressed at 5 dpi. Further, 334 host genes were differentially expressed compared with those before infection. Genes were clustered into four groups based on their expression profiles. Interferon-stimulated genes were more prevalent in groups showing upregulation at 5 dpi and 3 and 5 dpi. In contrast, the group showing downregulation at 3 dpi included inflammation-related genes, such as granzyme and eosinophil peroxidase genes. Downregulation of certain inflammation-related genes may contribute to the susceptibility of this fish to the virus.

Isolation and identification of a megalocytivirus strain (SKIV-TJ) from cultured spotted knifejaw (Oplegnathus punctatus) in China and its pathogenicity analysis

The spotted knifejaw (Oplegnathus punctatus) has recently emerged as a highly economically significant farmed fish in China. However, due to increasing environmental pollution and breeding density, a range of infectious diseases, including the iridovirus pathogen, have begun to spread widely. In this study, we isolated and identified a strain of Megalocytivirus, SKIV-TJ, from cultured spotted knifejaw in Tianjin, China. We observed significant cytopathic effects (CPE) in SKIV-TJ-infected spotted knifejaw brain (SKB) cells, and electron microscopy showed numerous virus particles in the cytoplasm of SKB cells 6 days post-infection. The annotated complete genome of SKIV-TJ (GenBank accession number ON075463) contained 112,489 bp and 132 open reading frames. Based on the multigene association evolutionary tree using 26 iridovirus core genes, SKIV-TJ was found to be most closely related to Rock bream iridovirus (RBIV). Cumulative mortality of spotted knifejaw infected with SKIV-TJ reached 100% by day 9. A transcriptomic analysis were conducted and a total of 5517 differentially expressed genes were identified, including 2757 upregulated genes and 2760 downregulated genes. The upregulated genes were associated with viral infection and immune signaling pathways. Our findings provide a valuable genetic resource and a deeper understanding of the immune response to SKIV infection in spotted knifejaw.

Characterization of Early Gonadal Differentiation and Induction of Sex Reversal in the Rock Bream Oplegnathus fasciatus.

Rock bream (Oplegnathus fasciatus) is a typical fish that has a unique multiple sex chromosome system (♀X1X1X2X2/♂X1X2Y). We examined the early gonadal development in rock bream via continuous histological observations of the gonads at 40-120days post hatching (dph). The fish was identified as a typical gonochorist, and female gonads were found to differentiate earlier than male gonads. The ovarian cavity of the female was initially observed at 80 dph, whereas the efferent duct of the male was not observed until 100 dph. Immunofluorescence with the vasa-antibody revealed that germ cells were predominantly distributed around the ovarian cavity in females and on the edge of the gonad in males during the early stages of sex differentiation. Sex reversal was induced via the oral administration of letrozole (LTZ), 17α-methyltestosterone (MT), and 17β-estradiol (E2), respectively, during the labile period of gonadal development. LTZ and MT induced 100% masculinization of genotype-females, whereas E2 induced only 50-60% feminization of genotype-males. Such findings suggest that the fish retained high sexual plasticity despite the existence of the neo-Y chromosome. MT and E2 had negative effect on fish growth, whereas LTZ did not exert such side effect. LTZ and MT could accelerate gonadal development in sex-reversed genotype-males, whereas E2 inhibited gonadal development in genotype-females of rock bream. These findings provide a basis for further research on the mechanisms of sex determination and differentiation in fishes with X1X2Y sex chromosome system and provide a sex reversal protocol for rock bream.

수온에 따른 참돔이리도바이러스(RSIV)의 감수성 품종에 대한 병원성 분석

Red sea bream iridovirus (RSIV) causes serious economic losses in Korea by causing mass mortality in rock bream fish during the season of high water temperatures (25~30℃). More than 30 additional species of cultured marine fish are listed as susceptible. Among them are major aquaculture species, including rock bream, red sea bream, black porgy, rockfish, and olive flounder. In this study, the effect of water temperature on the pathogenicity of RSIV in these fish species was investigated. RSIV was found to be pathogenic to rock bream at high temperatures (28℃) but not at low temperatures (18℃). In the histopathological investigation of the RSIV-infected rock bream at 18℃, only lymphocyte infiltration was seen, however after 14 days of infection at 28℃, development of inclusion body-bearing cells was observed. However, neither low nor high water temperatures showed RSIV genome copies, high cumulative mortality, or inclusion body-bearing cells in RSIV-infected black porgy, rockfish, or olive flounder. These results may contribute to selective quarantine measures based on pathogenicity and water temperature distinctions.

Red Sea Bream Iridovirus Kinetics, Tissue Tropism, and Interspecies Horizontal Transmission in Flathead Grey Mullets (Mugil cephalus).

Red sea bream iridovirus (RSIV) causes significant economic losses in the aquaculture industry. We analyzed the pathogenicity of RSIV in flathead grey mullets (Mugil cephalus), the correlation of histopathological lesions, and interspecies horizontal transmission, through immersion infection and cohabitation challenges. Flathead grey mullets, which were challenged by immersion infection, exhibited mortality at 14 and 24 days after RSIV exposure. Viral shedding in seawater peaked 2-3 days before or after the observed mortality. Specific lesions of RSIV were observed in the spleen and kidney, and the correlation between histopathological grade and viral load was the highest in the spleen. In a cohabitation challenge, flathead grey mullets were the donors, and healthy rock bream, red sea bream, and flathead grey mullets were the recipients. Viral shedding in seawater was the highest in flathead grey mullet and rock bream at 25 °C, with 106.0 RSIV copies L/g at 14 dpi. No mortality was observed in any group challenged at 15 °C, and no RSIV was detected in seawater after 30 dpi. The virus shed from RSIV-infected flathead grey mullets caused horizontal transmission through seawater. These findings suggest that rapid decision-making is warranted when managing disease in fish farms.

Impact of Red Sea Bream Iridovirus Infection on Rock Bream (Oplegnathus fasciatus) and Other Fish Species: A Study of Horizontal Transmission.

Red sea bream iridovirus (RSIV) causes significant economic losses in aquaculture. Here, we analyzed the pathogenicity, viral shedding, and transmission dynamics of RSIV in rock bream (Oplegnathus fasciatus) by employing immersion infection and cohabitation challenge models. Rock bream challenged by immersion exposure exhibited 100% mortality within 35 days post RSIV exposure, indicating that the viral shedding in seawater peaked after mortality. At 25 °C, a positive correlation between the viral loads within infected rock bream and virus shedding into the seawater was observed. Specific RSIV lesions were observed in the spleen and kidney of the infected rock bream, and the viral load in the spleen had the highest correlation with the histopathological grade. A cohabitation challenge mimicking the natural transmission conditions was performed to assess the virus transmission and determine the pathogenicity and viral load. The RSIV-infected rock breams (donors) were cohabited with uninfected rock bream, red sea bream (Pagrus major), and flathead grey mullet (Mugil cephalus) (recipients) at both 25 °C and 15 °C. In the cohabitation challenge group maintained at 15 °C, no mortality was observed across all experimental groups. However, RSIV was detected in both seawater and the recipient fish. Our results provide preliminary data for further epidemiological analyses and aid in the development of preventive measures and management of RSIVD in aquaculture.

A Comparative Study on Growth Performance, Body Composition, and Liver Tissue Metabolism Rearing on Soybean Lecithin-Enriched Artemia Nauplii and Microdiet in Rock Bream (Oplegnathus fasciatus) Larvae.

This study is aimed at establishing optimal soybean lecithin (SL) enrichment protocols in Artemia nauplii and at comparing the growth performance, body composition, and liver tissue metabolism in rock bream (Oplegnathus fasciatus) larvae reared on SL-enriched Artemia nauplii or SL-enriched microdiet (MD). The enrichment protocol experiment results indicated 12 h enrichment, and 10 g SL/m3 seawater could obtain desirable results. Rock bream larvae (25 days posthatching (dph)) were fed Artemia nauplii or MD for 30 days with three replicates. At stage 1 (larval 25-40 dph), significantly higher growth performance was observed in larvae fed the live prey (P < 0.05). Conversely, at stage 2 (41-55 dph), feeding with MD significantly increased larval standard length, and specific growth rate compared with those of larvae fed live prey. Larvae fed a MD showed decreased lipolysis-related lipase activity as well as decreased amino acid catabolism-related alanine aminotransferase and aspartate aminotransferase enzyme activities in liver tissue. RNA sequencing revealed that feeding with the MD primarily increased the expression of lipogenesis-related genes and protein translation-related gene expression in the liver tissue. Notably, feeding with MD significantly increased ribosome biogenesis-related genes as well as mitochondria synthesis-related gene expression, indicating a high protein anabolism rate and high energy production in liver tissue. In conclusion, 10 g SL/m3 seawater and 12 h could effectively enrich SL in Artemia nauplii. Retard weaning onto MD led to lower growth performance, which was likely due to the diversity of lipid and protein metabolism.

Structural insights on the selective interaction of the histidine-rich piscidin antimicrobial peptide Of-Pis1 with membranes

Of-Pis1 is a potent piscidin antimicrobial peptide (AMP), recently isolated from rock bream (Oplegnathus fasciatus). This rich in histidines and glycines 24-amino acid peptide displays high and broad antimicrobial activity and no significant hemolytic toxicity against human erythrocytes, suggesting low toxicity. To better understand the mechanism of action of Of-Pis1 and its potential selectivity, using NMR and CD spectroscopies, we studied the interaction with eukaryotic and procaryotic membranes and membrane models. Anionic sodium dodecyl sulfate (SDS) and lipopolysaccharide (LPS) micelles were used to mimic procaryotic membranes, while zwitterionic dodecyl phosphocholine (DPC) was used as eukaryotic membrane surrogate. In an aqueous environment, Of-Pis1 adopts a flexible random coil conformation. In DPC and SDS instead, the N-terminal region of Of-Pis1 forms an amphipathic α-helix with the non-polar face in close contact with the micelles. Slower solvent exchange and higher pKas of the histidine residues in SDS than in DPC suggest that Of-Pis1 interacts more tightly with SDS. Of-Pis1 also binds tightly and structurally perturbs LPS micelles. Of-Pis1 interacts with both Escherichia coli and mammalian cell membranes, but only in the presence of Escherichia coli membranes it populates the helical conformation. Furthermore, ligand-based NMR experiments support a tighter and more specific interaction with bacterial than with eukaryotic membranes. Overall, these data clearly show the selective interaction of this broadly active AMP with bacterial over eukaryotic membranes. The conformational information is discussed in terms of Of-Pis1 amino acid sequence and composition to provide insights useful to design more potent and selective AMPs.

Red Sea Bream Iridovirus (RSIV) Kinetics in Rock Bream (Oplegnathus fasciatus) at Various Fish-Rearing Seawater Temperatures.

Simple SummaryRed sea bream iridoviral disease (RSIVD) generates serious economic losses by causing mass mortality events of rock bream during the season with high water temperature in the Republic of Korea and other Asian countries. However, very few studies have investigated RSIV kinetics in rock bream under various rearing water temperatures. In this paper, we investigated the viral load shedding of RSIV into seawater after artificially infecting rock bream (Oplegnathus fasciatus) with the virus. Overall, our data suggest that the viral load shedding of RSIV into seawater varies depending on water temperature and virus inoculation concentration. Our results reveal the potential of non-invasive virus detection approaches, such as the utilization of environmental DNA in fish farms. In addition, we showed that the quantitative analysis of seawater viruses can indirectly improve our understanding of disease progression in fish, potentially contributing to enhanced disease control.Red sea bream iridoviral disease (RSIVD) causes serious economic losses in the aquaculture industry. In this paper, we evaluated RSIV kinetics in rock bream under various rearing water temperatures and different RSIV inoculation concentrations. High viral copy numbers (approximately 103.7–106.7 RSIV genome copies/L/g) were observed during the period of active fish mortality after RSIV infection at all concentrations in the tanks (25 °C and 20 °C). In the group injected with 104 RSIV genome copies/fish, RSIV was not detected at 21–30 days post-infection (dpi) in the rearing seawater. In rock bream infected at 15 °C and subjected to increasing water temperature (1 °C/d until 25 °C) 3 days later, the virus replication rate and number of viral copies shed into the rearing seawater increased. With the decrease in temperature (1 °C/d) from 25 to 15 °C after the infection, the virus replicated rapidly and was released at high loads on the initial 3–5 dpi, whereas the number of viral copies in the fish and seawater decreased after 14 dpi. These results indicate that the number of viral copies shed into the rearing seawater varies depending on the RSIV infection level in rock bream.

Effects of Soybean Lecithin on Growth Performance, Intestine Morphology, and Liver Tissue Metabolism in Rock Bream (Oplegnathus fasciatus) Larvae

Investigations have demonstrated a strong and positive association between dietary intact phospholipid (PL) inclusion and aquatic larval growth, nevertheless, the precise molecular mechanism underlying PL inclusion on growth performance has not been well elucidated. This study aimed to investigate the effects of dietary soybean lecithin (SL) inclusion on growth performance, liver metabolism, resistance to hypoxia stress, and potential molecular mechanisms in rock bream (Oplegnathus fasciatus) larvae. Four types of equal-protein and equal-lipid content microdiets (MDs) were formulated with graded levels of SL to achieve phospholipid levels of (PLs, dry matter) 3.84% (SL0), 6.71% (SL4), 9.38% (SL8), and 12.21% (SL12). Rock bream larvae (25 days post-hatching) were fed the respective MDs for 30 days with three replicates. We found that dietary SL inclusion promoted growth performance, survival rate, and stress resistance to hypoxia stress. The increased dietary SL inclusion improved intestinal structure, as shown by the increased perimeter ratio, muscular thickness, and mucosal fold height of the mid-intestinal tissue. Moreover, a high SL inclusion diet (SL12) increased the activity of the key lipolysis-related enzyme (lipase [LP]) in liver tissue but decreased the activity of amino acid catabolism-related enzymes (aspartate aminotransferase [AST] and alanine aminotransferase [ALT]). RNA sequencing results in liver tissue revealed that the SL12 diet increased the transcriptional level of fatty acid activation-related genes (acsl6 and acsbg2), phospholipid catabolism-related genes (acat2, lpin2, and crls), and amino acid synthesis-related genes (gs, csb, aldh18a1, and oct), but decreased the expression of amino acid catabolism-related gene gprt2. Notably, the SL12 diet significantly increased the expression of ribosome biogenesis-related genes (pes1, nop56, nop58, and rpf2) in liver tissue. The ribosome protein-related pathways were the most enriched pathways mapped in the GO database. Collectively, this study demonstrated the necessity of dietary SL for survival, growth performance, promotion of mid-intestinal morphology, and hypoxia stress during the rock bream larval stage. The SL-induced growth performance promotion was likely attributed to increasing nutrient acquisition by intestinal morphology improvement and to increasing SL catabolism and thereby sparing amino acids for protein synthesis.