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 ß), 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.

Saponin and chitosan-based oral vaccine against viral haemorrhagic septicaemia virus (VHSV) provides protective immunity in olive flounder (Paralichthys olivaceus).

Production losses of olive flounder (Paralichthys olivaceus) have increased owing to viral haemorrhagic septicaemia virus (VHSV) infection. In this study, we determined safe concentrations of orally administered saponin and chitosan by analysing serum enzyme (AST/ALT) levels as biochemical markers of hepatic injury. Furthermore, we demonstrated the efficacy, duration of protection, and safety of saponin and chitosan-based vaccines with inactivated VHSV (IV). Oral administration of saponin, chitosan, and their combination did not induce fish mortality at all tested concentrations (0.29, 1.45, and 2.9 mg/g of fish body weight/day) 10 days after administration. However, AST level was high at a dose >0.29 mg/g of fish body weight/day. Both saponin and chitosan were found to be safe and acceptable for vaccination studies at a dose of 0.29 mg/g of fish body weight/day. Administration of IV alone did not induce protection at 2 and 4 weeks post vaccination (wpv). Olive flounders administered saponin + IV and chitosan + IV vaccines had higher immunity against VHSV with relative percentage survival (RPS) of 12.5-7.5% and 0-20.1%, respectively; however, additional immunisation with combination of saponin + chitosan + IV clearly enhanced the protection with RPS values of 10-15%, 26.7%, 42.9%, and 37.5% at 4, 8, 12, and 20 wpv, respectively. Although the RPS value of oral immunisation was not comparable to that of injectable vaccines, the manufacturing process is simple and oral administration causes less stress to juvenile fish. To investigate the development of a protective immune response, olive flounder were re-challenged with VHSV (107.8 TCID50/fish) at 70 days postinfection; 100% of the previously unexposed fish died, whereas 80-100% of the previously immunised fish survived. Our results showed the possibility of developing preventive measures against VHSV using saponin and chitosan-based oral vaccines with inactivated virus.

Protective immunity induced by ankyrin repeat-containing protein-based DNA vaccine against rock bream iridovirus (RBIV) in rock bream (Oplegnathus fasciatus).

Rock bream iridovirus (RBIV) causes severe mass mortalities in rock bream (Oplegnathus fasciatus) and remains an unsolved problem in Korea aquaculture industry. In this study, we assessed the potential of ankyrin repeat (ANK)-containing proteins to induce protective immunity in RBIV-infected rock bream. Rock bream administered with ankyrin repeat-containing protein-based DNA vaccine (200 ng/fish) exhibited significant protection against at 4 and 8 weeks post vaccination to infected with 6.7 × 105 RBIV at 23°C; relative percent survival (RPS) of 60.04% and 40.1%, respectively. Furthermore, survivors from the first infection were strongly protected from RBIV (1.1 × 107) re-infection at 70 days post infection, as 100% RPS was observed and without clinical signs of RBIV diseases. Moreover, TLR3 (9.5-fold), TLR9 (5.2-fold), MyD88 (15.9-fold), Mx (55.5-fold), ISG15 (19.0-fold), PKR (24.2-fold), MHC class I (5.1-fold), perforin (6.5-fold), Fas (6.4-fold), Fas ligand (7.1-fold), caspase8 (5.0-fold), caspase9 (12.5-fold), and caspase3 (6.3-fold) responses were significantly elevated in the muscle (vaccine injection site) of ANK-based DNA vaccinated fish at 7 days post vaccination. However, inflammatory cytokines (IL1ß, IL8, and TNFα) were not enhanced in the vaccinated rock bream. Moreover, ANK gene may be a good candidate to detect RBIV infection or in revealing specific information to elucidate the pathogenic mechanisms underlying RBIV infection. In summary, ANK-based DNA vaccination in rock bream induced TLR- and IFN-mediated or apoptosis-related immune responses and suggest efficient preventive measures against RBIV.

Efficacy of saponin-based inactivated rock bream iridovirus (RBIV) vaccine in rock bream (Oplegnathus fasciatus).

Rock bream iridovirus (RBIV) causes severe mortality in rock bream (Oplegnathus fasciatus) for last two decades. In view of this constant threat of RBIV to the rock bream industry, we conducted the present study with the aim to develop a safe and efficient remedial measure against the virus. In this study, we evaluated the safety and potentiality of squalene, aluminium hydroxide and saponin adjuvants, singly or in combinations, which can be used for developing an efficient inactivated (IV) vaccine to protect rock bream from RBIV infection. The evaluation results demonstrated that saponin (Sa) has the required potential in enacting the antiviral immune response in the host and in providing protection against virus mediated lethality, without causing any adverted side-effects. The study further, showed that a single primary dose of Sa-adjuvanted IV vaccine can confer moderate protections in short (60.04% relative percent mortality (RPS) at 4 wpv) and medium (53.38% RPS at 8 wpv) term post RBIV challenge; whereas, the same vaccine when administered in a prime-boost strategy, it resulted enhanced 93.34% RPS post virus challenge at 4 and 8 wpv. The moderate to high survivability demonstrated by the Sa-adjuvanted IV vaccine, was substantiated by the significant (p < 0.05) upregulation of IL-1ß, Mx and PKR gene transcript. All surviving fish from the Sa-adjuvanted IV vaccine groups were strongly protected from re-infection with RBIV (1.1 × 107) at 70 days post infection (dpi). In conclusion, it can be inferred that, Sa-adjuvanted IV RBIV vaccine can be an efficient control measure to protect the rock bream aquaculture industry against the lethal RBIV virus.

Impact of Dark Excitons on the Population and Relaxation Kinetics of Two-Dimensional Biexcitons in [CH3(CH2)3NH3]2Pb1-xMnxBr4 (x = 0-0.09).

Two-dimensional (2D) semiconductors have emerged as an excellent platform for studying various excitonic matter under strong quantum and dielectric confinements. However, such effects can be seriously overestimated for Coulomb binding of two excitons to form a biexciton by a naive interpretation of the corresponding photoluminescence (PL) spectrum. By using 2D halide perovskite single crystals of [CH3(CH2)3NH3]2Pb1-xMnxBr4 (x = 0-0.09) as a model system, we investigated both population and relaxation kinetics of biexcitons as a function of excitation density, temperature, polarization, and Mn doping. We show that the biexciton is formed by binding of two dark excitons, which are partially bright, but they radiatively recombine to yield a bright exciton in the final state. This renders the spectral distance between the exciton peak and the biexciton peak as very different from the actual biexciton binding energy (ϕ) because of large bright-dark splitting. We show that Mn doping introduces paramagnetism to our 2D system and improves the biexciton stability as evidenced by increase in ϕ from 18.8 ± 0.7 to 20.0 ± 0.7 meV and the increase of the exciton-exciton capture coefficient C from 2.4 × 10-11 to 4.3 × 10-11cm2/ns within our doping range. The precisely determined ϕ values are significantly smaller than the previously reported ones, but they are consistent with the instability of the biexciton against thermal dissociation at room temperature. Our results demonstrate that electron-hole exchange interaction must be considered for precisely locating the biexciton level; therefore, the ϕ values should be reassessed for other 2D halide perovskites that even do not exhibit any dark exciton PL.

Orbit topology analyzed from π phase shift of magnetic quantum oscillations in three-dimensional Dirac semimetal.

With the emergence of Dirac fermion physics in the field of condensed matter, magnetic quantum oscillations (MQOs) have been used to discern the topology of orbits in Dirac materials. However, many previous researchers have relied on the single-orbit Lifshitz-Kosevich (LK) formula, which overlooks the significant effect of degenerate orbits on MQOs. Since the single-orbit LK formula is valid for massless Dirac semimetals with small cyclotron masses, it is imperative to generalize the method applicable to a wide range of Dirac semimetals, whether massless or massive. This report demonstrates how spin-degenerate orbits affect the phases in MQOs of three-dimensional massive Dirac semimetal, NbSb2 With varying the direction of the magnetic field, an abrupt π phase shift is observed due to the interference between the spin-degenerate orbits. We investigate the effect of cyclotron mass on the π phase shift and verify its close relation to the phase from the Zeeman coupling. We find that the π phase shift occurs when the cyclotron mass is half of the electron mass, indicating the effective spin gyromagnetic ratio as g s = 2. Our approach is not only useful for analyzing MQOs of massless Dirac semimetals with a small cyclotron mass but also can be used for MQOs in massive Dirac materials with degenerate orbits, especially in topological materials with a sufficiently large cyclotron mass. Furthermore, this method provides a useful way to estimate the precise g s value of the material.

Skyrmion Phase in MnSi Thin Films Grown on Sapphire by a Conventional Sputtering.

Topologically protected chiral skyrmions are an intriguing spin texture that has attracted much attention because of fundamental research and future spintronic applications. MnSi with a non-centrosymmetric structure is a well-known material hosting a skyrmion phase. To date, the preparation of MnSi crystals has been investigated by using special instruments with an ultrahigh vacuum chamber. Here, we introduce a facile way to grow MnSi films on a sapphire substrate using a relatively low vacuum environment of conventional magnetron sputtering. Although the as-grown MnSi films have a polycrystalline nature, a stable skyrmion phase in a broad range of temperatures and magnetic fields is observed via magnetotransport properties including phenomenological scaling analysis of the Hall resistivity contribution. Our findings provide not only a general way to prepare the materials possessing skyrmion phases but also insight into further research to stimulate more degrees of freedom in our inquisitiveness.

Signature of topological states in antiferromagnetic Sm-substituted Bi2Te3.

An antiferromagnetic topological insulator has been predicted to be preserved by breaking both time-reversal symmetry and primitive lattice translational symmetry. However, the topological surface state has often been observed to disappear in an antiferromagnetic phase because the doped magnetic impurity acts as an extrinsic defect. In this study, we report the experimental signature of topological surface states coexisting with antiferromagnetic order in Sm-doped Bi2Te3. We fabricate single crystals of SmxBi2-xTe3 with x = 0.004, 0.010, and 0.025, where the Curie-Weiss law is satisfied at low temperatures but is violated at high temperatures due to the influence of the high energy states of J multiplets of Sm. For x = 0.025, e xotic physical properties are observed, such as the antiferromagnetic phase with the Néel temperature TN = 3.3 K, multi-band Hall effect with two conduction channel, and anisotropic Shubnikov-de Haas oscillations. In the antiferromagnetic phase, we detect the signature of nontrivial topological surface states with surface electron density ns = 7.9 × 1011 cm-2 and its high mobility µs = 2,200 cm2/Vs, compared to nb = 2.0 × 1019 cm-3 and µb = 2.3 cm2/Vs for bulk electrons. These observations suggest that SmxBi2-xTe3 is a candidate creating the new stage for the potential application of topological antiferromagnetic spintronics.

Highly Reduced Saturation Magnetization in Epitaxially Grown Ferrimagnetic Heusler Thin Films.

The key of spintronic devices using the spin-transfer torque phenomenon is the effective reduction of switching current density by lowering the damping constant and the saturation magnetization while retaining strong perpendicular magnetic anisotropy. To reduce the saturation magnetization, particular conditions such as specific substitutions or buffer layers are required. Herein, we demonstrate highly reduced saturation magnetization in tetragonal D022 Mn3-x Ga thin films prepared by rf magnetron sputtering, where the epitaxial growth is examined on various substrates without any buffer layer. As the lattice mismatch between the sample and the substrate decreases from LaAlO3 and (LaAlO3)0.3(Sr2AlTaO6)0.7 to SrTiO3, the quality of Mn3-x Ga films is improved together with the magnetic and electronic properties. Especially, the Mn3-x Ga thin film epitaxially grown on the SrTiO3 substrate, fully oriented along the c axis perpendicular to the film plane, exhibits significantly reduced saturation magnetization as low as 0.06 µB, compared to previous results. By the structural and chemical analyses, we find that the predominant removal of Mn II atoms and the large population of Mn3+ ions affect the reduced saturation magnetization. Our findings provide insights into the magnetic properties of Mn3-x Ga crystals, which promise great potential for spin-related device applications.

Author Correction: Long-range chiral exchange interaction in synthetic antiferromagnets.

In the version of this Article originally published, the sentence 'D.-S.H. wrote the paper with K.L., J.H. and M.K.' in the author contributions was incorrect; it should have read 'D.-S.H. wrote the paper with K.L., J.H., M.-H.J. and M.K.' This has been corrected in the online versions of the Article.

Long-range chiral exchange interaction in synthetic antiferromagnets.

The exchange interaction governs static and dynamic magnetism. This fundamental interaction comes in two flavours-symmetric and antisymmetric. The symmetric interaction leads to ferro- and antiferromagnetism, and the antisymmetric interaction has attracted significant interest owing to its major role in promoting topologically non-trivial spin textures that promise fast, energy-efficient devices. So far, the antisymmetric exchange interaction has been found to be rather short ranged and limited to a single magnetic layer. Here we report a long-range antisymmetric interlayer exchange interaction in perpendicularly magnetized synthetic antiferromagnets with parallel and antiparallel magnetization alignments. Asymmetric hysteresis loops under an in-plane field reveal a unidirectional and chiral nature of this interaction, which results in canted magnetic structures. We explain our results by considering spin-orbit coupling combined with reduced symmetry in multilayers. Our discovery of a long-range chiral interaction provides an additional handle to engineer magnetic structures and could enable three-dimensional topological structures.

The Megalocytivirus RBIV Induces Apoptosis and MHC Class I Presentation in Rock Bream (Oplegnathus fasciatus) Red Blood Cells.

Rock bream iridovirus (RBIV) causes severe mass mortality in Korean rock bream (Oplegnathus fasciatus) populations. To date, immune defense mechanisms of rock bream against RBIV are unclear. While red blood cells (RBCs) are known to be involved in the immune response against viral infections, the participation of rock bream RBCs in the immune response against RBIV has not been studied yet. In this study, we examined induction of the immune response in rock bream RBCs after RBIV infection. Each fish was injected with RBIV, and virus copy number in RBCs gradually increased from 4 days post-infection (dpi), peaking at 10 dpi. A total of 318 proteins were significantly regulated in RBCs from RBIV-infected individuals, 183 proteins were upregulated and 135 proteins were downregulated. Differentially upregulated proteins included those involved in cellular amino acid metabolic processes, cellular detoxification, snRNP assembly, and the spliceosome. Remarkably, the MHC class I-related protein pathway was upregulated during RBIV infection. Simultaneously, the regulation of apoptosis-related proteins, including caspase-6 (CASP6), caspase-9 (CASP9), Fas cell surface death receptor (FAS), desmoplakin (DSP), and p21 (RAC1)-activated kinase 2 (PAK2) changed with RBIV infection. Interestingly, the expression of genes within the ISG15 antiviral mechanism-related pathway, including filamin B (FLNB), interferon regulatory factor 3 (IRF3), nucleoporin 35 (NUP35), tripartite motif-containing 25 (TRIM25), and karyopherin subunit alpha 3 (KPNA3) were downregulated in RBCs from RBIV-infected individuals. Overall, these findings contribute to the understanding of RBIV pathogenesis and host interaction.

Observation of Restored Topological Surface States in Magnetically Doped Topological Insulator.

The introduction of ferromagnetic order in topological insulators in general breaks the time-reversal symmetry and a gap is opened in topological surface bands. Various studies have focused on gap-opened magnetic topological insulators, because such modified band structures provide a promising platform for observing exotic quantum physics. However, the role of antiferromagnetic order in topological insulators is still controversial. In this report, we demonstrate that it is possible to restore the topological surface states by effectively reducing the antiferromagnetic ordering in Gd-substituted Bi2Te3. We successfully control the magnetic impurities via thermal treatments in ultra-high vacuum condition and observe apparent restoration of topological surface band dispersions. The microscopic mechanism of atomic rearrangements and the restoration process of topological surface states are unraveled by the combination of scanning tunneling microscopy measurements and density functional theory calculations. This work provides an effective way to control the magnetic impurities which is strongly correlated with topological surface states.

Temperature Dependence of the Dielectric Response and Critical Point Energies of Bi1.85Gd0.15Te3.

The complex dielectric function and band-edge critical point structures of Bi1.85Gd0.15Te3 are reported for temperatures from 28 to 300 K and energies from 0.74 to 6 eV, obtained on bulk Bi1.85Gd0.15Te3 by rotating-compensator spectroscopic ellipsometry. The critical point (CP) energies are determined using numerically calculated second energy derivatives of the data. At low temperature, eight CP structures are identified, while only four CPs are observed at room temperature. As temperature decreases, we also observe blue shifts and significantly enhanced CP structures relative to those obtained at room temperature. The temperature dependence of the CPs are determined by fitting the data to the temperature coefficient and a phenomenological expression that contains the Bose-Einstein statistical factor.

DNA vaccine encoding myristoylated membrane protein (MMP) of rock bream iridovirus (RBIV) induces protective immunity in rock bream (Oplegnathus fasciatus).

Rock bream iridovirus (RBIV) causes severe mass mortalities in rock bream (Oplegnathus fasciatus) in Korea. In this study, we investigated the potential of viral membrane protein to induce antiviral status protecting rock bream against RBIV infection. We found that fish administered with ORF008L (myristoylated membrane protein, MMP) vaccine exhibited significantly higher levels of survival compared to ORF007L (major capsid protein, MCP). Moreover, ORF008L-based DNA vaccinated fish showed significant protection at 4 and 8 weeks post vaccination (wpv) than non-vaccinated fish after infected with RBIV (6.7 × 105) at 23 °C, with relative percent survival (RPS) of 73.36% and 46.72%, respectively. All of the survivors from the first RBIV infection were strongly protected (100% RPS) from re-infected with RBIV (1.1 × 107) at 100 dpi. In addition, the MMP (ORF008L)-based DNA vaccine significantly induced the gene expression of TLR3 (14.2-fold), MyD88 (11.6-fold), Mx (84.7-fold), ISG15 (8.7-fold), PKR (25.6-fold), MHC class I (13.3-fold), Fas (6.7-fold), Fas ligand (6.7-fold), caspase9 (17.0-fold) and caspase3 (15.3-fold) at 7 days post vaccination in the muscle (vaccine injection site). Our results showed the induction of immune responses and suggest the possibility of developing preventive measures against RBIV using myristoylated membrane protein-based DNA vaccine.

Efficacy of algal Ecklonia cava extract against viral hemorrhagic septicemia virus (VHSV).

The inhibition efficacy of an extract from Ecklonia cava (E. cava) was studied to determine whether the extract and compounds exhibited inhibitory activity against VHSV in the fathead minnow (FHM) cell line and following oral administration to the olive flounder. Based on its low toxicity and effective concentration, the E. cava extract (Ext) and compounds (eckol and phlorofucofuroeckol A) were selected for further analysis. In the plaque reduction assay, simultaneous co-exposure of VHSV to Ext, eckol and phlorofucofuroeckol A showed a higher level of inhibition than the pre- and post-exposure groups. The antiviral activity in the FHM cell line was time-dependent and increased with the exposure time with the virus and Ext or the compounds. In the in vivo experiments, different Ext concentrations were orally administered to the olive flounder. In trial I, the relative percent survival (RPS) following oral administration of 500 and 50 µg/g/day of Ext was 31.25% and 12.50%, respectively. In trial II, the RPS for 1000, 500 and 50 µg/g/day of Ext was 31.57%, 0% and 0%, respectively. In trial III, the RPS after 1 and 2 weeks (1000 µg/g/day) of exposure to Ext was 26.31% and 31.57%, respectively. Oral administration of Ext (1000 µg/g/day) significantly induced inflammatory cytokine responses (IL-1ß, IL-6 and IFN-γ) at 1 and 2 days post-oral administration (dpa). Additionally, IFN-α/ß (7-12 dpa), ISG15 (2, 7 and 10 dpa) and Mx (7-12 dpa) were significantly activated in the olive flounder. In conclusion, we demonstrated an inhibitory ability of the E. cava extract and compounds against VHSV in the FHM cell line. Moreover, oral administration of the E. cava extract to the olive flounder enhanced antiviral immune responses and the efficacy of protection against VHSV, resulting in an anti-viral status in the olive flounder.

Innate immune responses against rock bream iridovirus (RBIV) infection in rock bream (Oplegnathus fasciatus) following poly (I:C) administration.

Poly (I:C) showed promise as an immunoprotective agents in rock bream against rock bream iridovirus (RBIV) infection. In this study, we evaluated the time-dependent virus replication pattern and antiviral immune responses in RBIV-infected rock bream with and without poly (I:C) administration. In the poly (I:C)+virus-injected group, virus copy numbers were more than 18.9-, 24.0- and 479.2-fold lower than in the virus only injected group at 4 (4.73 × 104 and 8.95 × 105/µl, respectively), 7 (3.67 × 105 and 8.81 × 106/µl, respectively) and 10 days post infection (dpi) (1.26 × 105 and 6.02 × 107/µl, respectively). Moreover, significantly high expression levels of TLR3 (8.6- and 7.7-fold, at 4 and 7 dpi, respectively) and IL1ß (3.6-fold at 2 dpi) were observed in the poly (I:C)+virus-injected group, but the expression levels were not significantly in the virus-injected group. However, IL8 and TNFα expression levels showed no statistical significance in both groups. Mx, ISG15 and PKR were significantly highly expressed from 4 to 10 dpi in the virus-injected group. Nevertheless, in the poly (I:C)+virus-injected group, Mx and ISG15 expression were significantly expressed from 2 dpi. In summary, poly (I:C) administration in rock bream induces TLR3, IL1ß, Mx and ISG15-mediated immune responses, which could be a critical factor for inhibition of virus replication.

Rock bream iridovirus (RBIV) replication in rock bream (Oplegnathus fasciatus) exposed for different time periods to susceptible water temperatures.

Rock bream iridovirus (RBIV) is a member of the Megalocytivirus genus that causes severe mortality to rock bream. Water temperature is known to affect the immune system and susceptibility of fish to RBIV infection. In this study, we evaluated the time dependent virus replication pattern and time required to completely eliminate virus from the rock bream body against RBIV infection at different water temperature conditions. The rock bream was exposed to the virus and held at 7 (group A1), 4 (group A2) and 2 days (group A3) at 23 °C before the water temperature was reduced to 17 °C. A total of 28% mortality was observed 24-35 days post infection (dpi) in only the 7 day exposure group at 23 °C. In all 23 °C exposure groups, virus replication peaked at 20 to 22 dpi (106-107/µl). In recovery stages (30-100 dpi), the virus copy number was gradually reduced, from 106 to 101 with faster decreases in the shorter exposure period group at 23 °C. When the water temperature was increased in surviving fish from 17 to 26 °C at 70 dpi, they did not show any mortality or signs of disease and had low virus copy numbers (below 102/µl). Thus, fish need at least 50 days from peaked RBIV levels (approximately 20-25 dpi) to inhibit the virus. This indicates that maintaining the fish at low water temperature (17 °C) for 70 days is sufficient to eradicate RBIV from fish body. Thus, RBIV could be eliminated slowly from the fish body and the virus may be completely eliminated under the threshold of causing mortality.

Gene expression regulation of the TLR9 and MyD88-dependent pathways in rock bream against rock bream iridovirus (RBIV) infection.

Rock bream iridovirus (RBIV), which is a member of the Megalocytivirus genus, causes severe mass mortalities in rock bream in Korea. To date, the innate immune defense mechanisms of rock bream against RBIV is unclear. In this study, we assessed the expression levels of genes related to TLR9 and MyD88-dependent pathways in RBIV-infected rock bream in high, low or no mortality conditions. In the high mortality group (100% mortality at 15 days post infection (dpi)), high levels of TLR9 and MyD88 expressions (6.4- and 2.4-fold, respectively) were observed at 8 d and then reduced (0.6- and 0.1-fold, respectively) with heavy viral loads at 10 dpi (2.21 × 107/µl). Moreover, TRAF6, IRF5, IL1ß, IL8, IL12 and TNFα expression levels showed no statistical significance until 10 dpi. Conversely, in the low mortality group (28% expected mortality at 35 dpi), TLR9, MyD88 and TRAF6 expression levels were significantly higher than those in the control group at several sampling points until 30 dpi. Higher levels of IRF5, IL1ß, IL8, IL12 and TNFα expression were also observed, however, these were not significantly different from those of the control group. In the no mortality group (0% mortality at 40 dpi), significantly higher levels of MyD88 (2 d, 4 d and 40 dpi), TRAF6 (2 dpi), IL1ß (4 dpi) and IL8 (2 d and 4 dpi) expression were observed. In summary, RBIV-infected rock bream induces innate immune response, which could be a major contributing factor to effective fish control over viral transcription. MyD88, TRAF6, IL1ß and IL8-related immune responses were activated in fish survivor condition (low or no mortality group). This is a critical factor for RBIV disease recovery; however, these immune responses did not efficiently respond in fish dead condition (high mortality group).

CpG ODN 1668 induce innate and adaptive immune responses in rock bream (Oplegnathus fasciatus) against rock bream iridovirus (RBIV) infection.

Rock bream iridovirus (RBIV) causes severe mass mortalities in rock bream in Korea. CpG ODN 1668 showed promise as immunoprotective agents against RBIV infection in rock bream. In this study, we assessed innate/adaptive-related gene expression patterns in RBIV-infected rock bream with and without CpG ODN 1668 administration to determine important immune defense related factors that may affect fish survival. In the CpG ODN 1668+virus-injected group, virus copies were more than 7.4- to 790591-fold lower than in the virus-injected group at 4 d (8.79 × 104 and 6.58 × 105/µl, respectively), 7 d (5.30 × 102 and 2.29 × 107/µl, respectively) and 10 dpi (7.79 × 101 and 6.16 × 107/µl, respectively). Furthermore, in the CpG ODN 1668+virus-injected group, significantly higher levels of MyD88 (6 h, 1 d, 4 d and 7 dpi), IL1ß (1 d, 2 d and 7 dpi) and perforin/granzyme (1 dpi) expression were observed, whereas these genes were not significantly expressed in the virus-injected group at that time points. Mx, ISG15 and PKR were significantly highly expressed at 4 d and 7 dpi and reduced when low viral loads at 10 dpi in the CpG ODN 1668+virus-injected group. Conversely, in the virus-injected group, Mx, ISG15 and PKR expression were significantly higher than the control group until 10 dpi. However, MHC class I, CD8, Fas, Fas ligand and caspases (3, 8 and 9) expression levels showed no statistically significant differences between virus- and CpG ODN 1668+virus-injected group. In summary, CpG ODN 1668 administration in fish induces innate immune response or cell death pathway, which could be a major contributing factor to effective fish control over viral transcription on 4 d to 10 dpi. Expression of MyD88, IL1ß, perforin and granzyme-related immune gene response is critical factor for inhibition of RBIV replication.