Epinephelus Coioides Research Articles

Fish ELOVL7a is involved in virus replication via lipid metabolic reprogramming

The elongation of very long chain fatty acids (ELOVL) proteins are key rate-limiting enzymes that catalyze fatty acid synthesis to form long chain fatty acids. ELOVLs also play regulatory roles in the lipid metabolic reprogramming induced by mammalian viruses. However, little is known about the roles of fish ELOVLs during virus infection. Here, a homolog of ELOVL7 was cloned from Epinephelus coioides (EcELOVL7a), and its roles in red-spotted grouper nervous necrosis virus (RGNNV) and Singapore grouper iridovirus (SGIV) infection were investigated. The transcription level of EcELOVL7a was significantly increased upon RGNNV and SGIV infection or other pathogen-associated molecular patterns stimulation in grouper spleen (GS) cells. Subcellular localization analysis showed that EcELOVL7a encoded an endoplasmic reticulum (ER) related protein. Overexpression of EcELOVL7a promoted the viral production and virus release during SGIV and RGNNV infection. Furthermore, the lipidome profiling showed that EcELOVL7a overexpression reprogrammed cellular lipid components in vitro, evidenced by the increase of glycerophospholipids, sphingolipids and glycerides components. In addition, VLCFAs including FFA (20:2), FFA (20:4), FFA (22:4), FFA (22:5) and FFA (24:0), were enriched in EcELOVL7a overexpressed cells. Consistently, EcELOVL7a overexpression upregulated the transcription level of the key lipid metabolic enzymes, including fatty acid synthase (FASN), phospholipase A 2α (PLA 2α), and cyclooxygenases −2 (COX-2), LPIN1, and diacylglycerol acyltransferase 1α (DGAT1α). Together, our results firstly provided the evidence that fish ELOVL7a played an essential role in SGIV and RGNNV replication by reprogramming lipid metabolism.

Slc43a2+ T cell metastasis from spleen to brain in RGNNV infected teleost.

The origin of T cells in the teleost's brain is unclear. While viewing the central nervous system (CNS) as immune privileged has been widely accepted, previous studies suggest that T cells residing in the thymus but not in the spleen of the teleost play an essential role in communicating with the peripheral organs. Here, we identified nine T cell subpopulations in the thymus and spleen of orange-spotted grouper (Epinephelus coioices) through single-cell RNA-sequencing analysis. After viral CNS infection with red-spotted grouper nervous necrosis virus (RGNNV), the number of slc43a2+ T cells synchronously increased in the spleen and brain. During the infection tests in asplenic zebrafish (tlx1▲ zebrafish model), no increase in the number of slc43a2+ T cells was observed in the brain. Single-cell transcriptomic analysis indicated that slc43a2+ T cells mature and functionally differentiate within the spleen and then migrate into the brain to trigger an immune response. This study suggests a novel route for T cell migration from the spleen to the brain during viral infection in fish.

The effects of four paralogous piscidin antimicrobial peptides on the chemotaxis, macrophage respiratory burst, phagocytosis and expression of immune-related genes in orange-spotted grouper (Epinephelus coicodes)

Antimicrobial peptides (AMPs) are an essential part of the vertebrate innate immune system. Piscidins are a family of AMPs specific in fish. In our previous investigation, we identified four paralogous genes of piscidins in the orange-spotted grouper (Epinephelus coicodes), which exhibited distinct activities against bacteria, fungi, and parasitic ciliated protozoa. Piscidins demonstrated their capability to modulate the expression of diverse immune-related genes; however, their precise immunoregulatory functions remain largely unexplored. In this study, we examined the immunomodulatory properties of putative mature peptides derived from four E. coicodes piscidins (ecPis1S, ecPis2S, ecPis3S, and ecPis4S) in head kidney leukocytes (HKLs) or monocytes/macrophages (MO/MΦ)-like cells isolated from E. coicodes. Our data demonstrate that E. coicodes piscidins exhibit immunomodulatory activities supported by multiple lines of evidence. Firstly, all four piscidins displayed chemotactic activities towards HKLs, with the most potent chemotactic activity observed in ecPis2S. Secondly, stimulation with E. coicodes piscidins enhanced respiratory burst and phagocytic activity in MO/MФ-like cells, with ecPis3S showing the highest efficacy in increasing phagocytosis of MO/MΦ-like cells. Thirdly, mRNA expression levels of chemokine receptors, Toll-like receptors, T cell receptors, and proinflammatory cytokines were modulated to varying extents by the four piscidins in E. coicodes HKLs. Overall, our findings indicate that the immunological activities of these four paralogous piscidins from E. coicodes are exhibited in a paralog-specific and concentration-dependent manner, highlighting their distinct and versatile immunomodulatory properties. This study makes a significant contribution to the field of fish AMPs immunology by elucidating the novel mechanisms through which members of the piscidin family exert their immunomodulatory effects. Moreover, it provides valuable insights for further exploration of fish immunomodulating agents.

Characterization and functional analysis of SOCS9 from orange-spotted grouper (Epinephelus coioides) during virus infection

The suppressor of cytokine signaling (SOCS) proteins family have twelve members including eight known mammalian SOCS members (CISH, SOCS1-7) and four new discovery members (SOCS3b, SOCS5b, SOCS8 and SOCS9) that is regarded as a classic feedback inhibitor of cytokine signaling. Although the function of the mammalian SOCS proteins have been well studied, little is known about the roles of SOCS in fish during viral infection. In this study, the molecular characteristics of SOCS9 from orange-spotted grouper (Epinephelus coioides, EcSOCS9) is investigated. The EcSOCS9 protein encoded 543 amino acids with typical SH2 (389-475aa) and SOCS_box (491-527aa), sharing high identities with reported fish SOCS9. EcSOCS9 was expressed in all detected tissues and highly expressed in kidney. After red-spotted grouper nervous necrosis virus (RGNNV) infection, the expression of EcSOCS9 was significantly induced in vitro. Furthermore, EcSOCS9 overexpression enhanced RGNNV replication, promoted virus-induced mitophagy that evidenced by the increased level of LC3-Ⅱ, BCL2, PGAM5 and decreased level of BNIP3 and FUNDC1. Besides, EcSOCS9 overexpression suppressed the expression levels of ATP6, CYB, ND4, ATP level and induced ROS level. The expression levels of interferon (IFN) related factors (IRF1, IRF3, IRF7, P53), inflammatory factors (IL1-β, IL8, TLR2, TNF-α) and IFN-3, ISRE, NF-κB, AP1 activities were also reduced by overexpressing EcSOCS9. These date suggests that EcSOCS9 impacts RGNNV infection through modulating mitophagy, regulating the expression levels of IFN- related and inflammatory factors, which will expand our understanding of fish immune responses during viral infection.

Orange-spotted grouper IFNh response to NNV or MSRV and its potential antiviral activities

Type I interferon (IFN) plays a crucial role in the antiviral immune response. Nervous necrosis virus (NNV) and Micropterus salmoides rhabdovirus (MSRV) are the most important viruses in cultured larvae and juveniles, causing great economic losses to fish farming. To better understand the antiviral activities and immunoregulatory role of IFN from orange-spotted grouper (Epinephelus coioides), EcIFNh was cloned from NNV infected sample. EcIFNh has an open reading frame (ORF) of 552 bp and encodes a polypeptide of 183 amino acids. Phylogenetic tree analysis showed that EcIFNh was clustered into the IFNh branch. The tissue distribution analysis revealed that EcIFNh was highly expressed in the liver and brain of healthy orange-spotted grouper. The mRNA levels of EcIFNh were significantly upregulated after poly (I:C) stimulation and NNV or MSRV infection. Furthermore, the promoter of EcIFNh was characterized and significantly activated by EcMDA5, EcMAVS, EcSTING, EcIRF3, and EcIRF7 in the luciferase activity assays. We found that EcIFNh overexpression resisted the replication of NNV and MSRV, while EcIFNh silencing facilitated NNV replication in GB cells. In addition, EcIFNh recombinant protein (rEcIFNh) enhanced the immune response by inducing the expression of ISGs in vivo and in vitro, suggesting the potential application of rEcIFNh for anti-NNV and anti-MSRV. Taken together, our research may offer the foundation for virus-IFN system interaction in orange-spotted grouper.

Functional properties of ATPIF1 in the orange-spotted grouper (Epinephelus coioides) in response to viral infection

ATP synthase inhibitory factor 1 (ATPIF1) can activate mitochondrial autophagic pathway and mediates immune response by regulating ATP synthase activity. However, the role of fish ATPIF1 on viral infection is still unknown. In this study, we identified an ATPIF1 homolog (Ec-ATPIF1) from orange-spotted grouper (Epinephelus coioides). Ec-ATPIF1 is mainly expressed in the kidney and liver. The expression of Ec-ATPIF1 was significantly up-regulated after RGNNV stimulation in vitro. Further experiments showed that overexpression of Ec-ATPIF1 inhibited the expression of viral genes (CP and RdRp) and intracellular ATP synthesis. Ec-ATPIF1 overexpression also promoted the expression of mitophagy related genes (PINK1, Parkin, BNIP3, NIX, FUNDC1, LC3), inflammation-related factors (IL-1β, IL-6, IL-8, IL-10, TNF-α, TLR2) and interferon pathway factors (IRF1, IRF3, IRF7, MX1, ISG15, ISG56, MDA5, TRIF). While the knockdown of Ec-ATPIF1 exhibited the opposite effects on the expression of viral genes and immune-related factors above. These data suggest that Ec-ATPIF1 can impact viral infection by regulating mitophagy, ATP synthesis, the expression of inflammatory factors and interferon pathway factors. These findings will be beneficial to better explore the immune regulatory mechanisms of fish respond to viral infection.

Untangling the Complexity of Small-Scale Fisheries: Building an Understanding of Grouper-Snapper Fisheries Dynamics in Saleh Bay, West Nusa Tenggara, Indonesia

Small-scale tropical fisheries are complex systems that utilize multiple fishing gears to target various species. In this study, we investigated small-scale reef and demersal fisheries in Saleh Bay, Indonesia, using catch and effort data from 2016 to 2019, where 57.7 percent of the catch was grouper (serranids) and snapper (lutjanids). Despite the complexity (75 documented species and eight fishing methods), this fishery is characterized by the catches of four dominant species: leopard coral grouper (Plectropomus leopardus), orange-spotted-grouper (Epinephelus coioides), spotted coral grouper (P. maculatus), and malabar blood snapper (Lutjanus malabaricus). The species caught varied among fishing methods. Over 90% of the catch was attributed to three primary fishing methods: bottom longline, speargun, and handline. Multivariate analyses found that fishing depth, season, and/or year significantly influenced the catch composition for each of these fishing methods. Fishing activities exhibit a temporal pattern influenced by monsoonal seasons. Results also suggest that fishers employ specialized fishing tactics by targeting high-value species to maximize their profits. This study recommends a management strategy of focusing on monitoring and managing the three main fishing gears and four important species during their peak seasons to reduce some of the complexity and management costs.

Dual-specificity phosphatase 1 inhibits Singapore grouper iridovirus replication via regulating apoptosis in Epinephelus coioides

The dual-specificity phosphatase (DUSP) family plays key roles in the maintenance of cellular homeostasis and apoptosis etc. In this study, the DUSP member DUSP1 of Epinephelus coioides was characterized: the length was 2371 bp including 281 bp 5′ UTR, 911 bp 3′ UTR, and a 1125 bp open reading frame encoding 374 amino acids. E. coioides DUSP1 has two conserved domains, a ROHD and DSPc along with a p38 MAPK phosphorylation site, localized at Ser308. E. coioides DUSP1 mRNA can be detected in all of the tissues examined, and the subcellular localization showed that DUSP1 was mainly distributed in the nucleus. Singapore grouper iridovirus (SGIV) infection could induce the differential expression of E. coioides DUSP1. Overexpression of DUSP1 could inhibit SGIV-induced cytopathic effect (CPE), the expressions of SGIV key genes, and the viral titers. Overexpression of DUSP1 could also regulate SGIV-induced apoptosis, and the expression of apoptosis-related factor caspase 3. The results would be helpful to further study the role of DUSP1 in viral infection.

Function analysis of fish PACT gene in response to virus infection

PACT (interferon-inducible double-stranded RNA-dependent protein kinase activator A) is a cellular protein which can activate PKR in dsRNA-independent manner. However, the role of PACT in fish virus infection remains largely unknown. In this study, a PACT homologue from grouper (Epinephelus coioides)(EcPACT) was cloned and characterized. The open reading frame of EcPACT has a full length of 924 bp and encodes a protein of 307 amino acids with a predicted molecular weight of 33.29 kDa. Similar to mammals, EcPACT contains three dsRBD domains. EcPACT shares 99.67 % homology with E. lanceolatus. Real-time fluorescence quantitative PCR results showed that EcPACT mRNA was widely expressed in all tissues and abundantly expressed in brain, blood, head kidney and kidney. In addition, SGIV and RGNNV infection significantly upregulated the transcript levels of EcPACT. Subcellular localization analysis showed that EcPACT was mainly distributed in the nucleus. Overexpression of EcPACT inhibited the replication of SGIV and RGNNV in vitro and positively regulated the expression of interferon (IFN) and pro-inflammatory factors. The results provide a better understanding of the relationship between PACT and viral infection in fish.

UNC93B1 facilitates the localization and signaling of TLR5M in Epinephelus coioides

Toll-like receptor 5 (TLR5), serving as a sensor of bacterial flagellin, mediates the innate immune response to actively engage in the host's immune processes against pathogen invasion. However, the mechanism underlying TLR5-mediated immune response in fish remains unclear. Despite the presumed cell surface expression of TLR5 member form (TLR5M), its trafficking dynamics remain elusive. Here, we have identified Epinephelus coioides TLR5M as a crucial mediator of Vibrio flagellin-induced cytokine expression in grouper cells. EcTLR5M facilitated the activation of NF-κB signaling pathway in response to flagellin stimulation and exerted a modest influence on the mitogen-activated protein kinase (MAPK)-extracellular regulated kinase (ERK) signaling. The trafficking chaperone Unc-93 homolog B1 (EcUNC93B1) participated in EcTLR5M-mediated NF-κB signaling activation and downstream cytokine expression. In addition, EcUNC93B1 combined with EcTLR5M to mediate its exit from the endoplasmic reticulum, and also affected its post-translational maturation. Collectively, these findings first discovered that EcTLR5M mediated the flagellin-induced cytokine expression primarily by regulating the NF-κB signaling pathway, and EcUNC93B1 mediated EcTLR5M function through regulating its trafficking and post-translational maturation. This research expanded the understanding of fish innate immunity and provided a novel concept for the advancement of anti-vibrio immunity technology.

DNA Barcoding of Six Commercially Important Groupers (Epinephelidae) from Langsa, Aceh, Indonesia

Groupers are among the fish groups that are difficult to recognize due to their high morphological similarities. Therefore, molecular techniques, particularly DNA barcoding, are extensively utilized to differentiate this fish group. This study aimed to analyze and validate six grouper species belonging to the Epinephelidae family that were harvested from Langsa district waters in Aceh province, Indonesia, based on DNA barcode data. It was conducted from June to December 2021, with the fish specimens collected from fishers at fish landing sites and the fish market in Langsa City, Aceh province. A total of 22 grouper sequences belonging to six species were generated, namely Epinephelus coioides, E. bleekeri, E. malabaricus, E. erythrurus, E. sexfasciatus and Mycteroperca poecilonotus (formerly Epinephelus poecilonotus). Genetic distance within these species ranged from 0.10 to 0.73% (average: 0.40%). Notably, E. malabaricus and E. coioides exhibited the closest genetic kinship (4.07%), while E. sexfasciatus and M. poecilonotus displayed the greatest genetic distance (19.33%). This study provides the first DNA reference for grouper in Langsa district, Indonesia, with significant implications for future sustainable grouper management.

PENDUGAAN STATUS STOK DAN INDIKATOR BIOEKONOMI PERIKANAN KERAPU DI TELUK SALEH, NUSA TENGGARA BARAT

One of the challenges in managing small-scale fisheries with limited data is the need for indicators to estimate the stock status and the total allowable catch for sustainable utilization of fish resources. This study aims to estimate the stock status, catch, and optimal fishing effort of the grouper fishery in Saleh Bay, West Nusa Tenggara Province by using the Length-based Spawning Potential Ratio analysis, production surplus model, and bioeconomic approach. Based on the results of this study, it is known that Plectopomus leopardus and Epinephelus coioides are in overexploited condition, while Plectropomus maculatus is in fully exploited status. This study also shows that with a bioeconomic approach, optimal conditions for grouper fisheries in these waters are achieved when the catch is 3,862.1 tonnes per year. The results of this study underline the importance of maximum economic yield-based management as a framework for a fishing quota-based system as an additional instrument in implementing sustainable fishery management in the future. The success of applying the quota system and the total allowable effort is determined by the accuracy of the catch data of small vessels around Saleh Bay under regulatory provisions, supervision and effective law enforcement, the socio-cultural conditions of local communities and the level of compliance with regulations.Keywords: fishing effort, grouper, overfished, quota system.

Molecular and Functional Characterization of α Chain of Interleukin-15 Receptor (IL-15Rα) in Orange-Spotted Grouper (Epinephelus coioides) in Response to Vibrio harveyi Challenge

Molecular and Functional Characterization of α Chain of Interleukin-15 Receptor (IL-15Rα) in Orange-Spotted Grouper (Epinephelus coioides) in Response to Vibrio harveyi Challenge

Effect of pH, osmolality, Na+ and K+ on spermatozoa motility of the orange-spotted grouper, Epinephelus coioides

ABSTRACT The present study investigated the biosensitivity of Epinephelus coioides spermatozoa under various ionic and osmolality conditions. Activation and sensitivity of sperm started at different pH (7, 7.5, 8, 8.5, and 9) level. The highest motility duration was obtained in pH 8.5 with a significant difference to other groups (P < .001). The E. coioides spermatozoa exposed to 600 mM NaCl solution, 300 mM KCl (in the medium containing 200 mM sucrose), and sucrose solutions with 1100 mosmol/kg osmolality had significantly higher duration of motility than those exposed to solutions with the lower amount of NaCl, KCl and osmolality. Any amount greater than 600 mM NaCl, 300 mM KCl and 1100 mosmol/kg osmolality showed a remarkable reduction in the motility of spermatozoa. Results also showed that the amount of 100 and 450 mM K+, and osmolality lower than 400 mosmol/kg had a significant inhibitory effect and prevented the activity of spermatozoa. The findings of this study showed that changes in K+, Na+ and osmolality of the external solutions affects the spermatozoa and its motility. The potential use of these ions could be applicable for the improvement of the spermatozoa quality of E. coioides during artificial reproduction.

Rab32, a novel Rab small GTPase from orange-spotted grouper, Epinephelus coioides involved in SGIV infection

Rab32 is a member of the Rab GTPase family that is involved in membrane trafficking and immune response, which are crucial for controlling pathogen infection. However, the role of Rab32 in virus infection is not well understood. In this study, we focused on the regulation of Rab32 on virus infection and the host immunity in orange-spotted grouper, Epinephelus coioides. EcRab32 encoded a 213-amino acid polypeptide, which shared a high sequence identity with other Rab32 proteins from fishes to mammals. In healthy orange-spotted grouper, the mRNA of EcRab32 was expressed in all the detected tissues, with the more expression levels in the head kidney, liver and gill. Upon SGIV infection, the expression of EcRab32 was significantly up-regulated in vitro, indicating its potential role in viral infection. EcRab32 was observed to be distributed in the cytoplasm as punctate and vesicle-like structures. EcRab32 overexpression was found to notably inhibit SGIV infection, while the interruption of EcRab32 significantly promoted SGIV infection. In addition, using single particle imaging analysis, we found that EcRab32 overexpression prominently reduced the attachment and internalization of SGIV particles. Furthermore, the results demonstrated that EcRab32 played a positive role in regulating the interferon immune and inflammatory responses. Taken together, these findings indicated that EcRab32 influenced SGIV infection by regulating the host immune response, providing an overall understanding of the interplay between the Rab32 and innate immunity.

An Integrated in silico and in vivo study of nucleic acid vaccine against Nocardia seriolae infection in orange-spotted grouper Epinephelus coioides

Nocardiosis in aquatic animals caused by Nocardia seriolae is a frequently occurring serious infection that has recently spread to many countries. In this study, DNA vaccines containing potential bacterial antigens predicted using the reverse vaccinology approach were developed and evaluated in orange-spotted groupers. In silico analysis indicated that proteins including cholesterol oxidase, ld-transpeptidase, and glycosyl hydroxylase have high immunogenicity and are potential vaccine candidates. In vitro assays revealed the mature and biological configurations of these proteins. Importantly, when compared to a control PBS injection, N. seriolae DNA-based vaccines showed significantly higher expression of IL1β, IL17, and IFNγ at 1 or 2 days, in line with higher serum antibody production and expression of other cellular immune-related genes, such as MHCI, CD4, and CD8, at 7 days post-immunization. Remarkably, enhanced immune responses and strong protective efficacy against a highly virulent strain of N. seriolae were recorded in DNA vaccine-cholesterol oxidase (pcD::Cho) injected fish, with a relative survival rate of 73.3%. Our results demonstrate that the reverse vaccinology approach is a valid strategy for screening vaccine candidates and pcD::Cho is a promising candidate that can boost both innate and adaptive immune responses and confer considerable protection against N. seriolae infection.

Grouper Rab1 inhibits nodovirus infection by affecting virus entry and host immune response

Rab1, a GTPase, is present in all eukaryotes, and is mainly involved in vesicle trafficking between the endoplasmic reticulum and Golgi, thereby regulating many cellular activities and pathogenic infections. However, little is known of how Rab1 functions in fish during virus infection. Groupers (Epinephelus spp.) are high in economic value and widely cultivated in China and Southeast Asia, although they often suffer from diseases. Red-spotted grouper nervous necrosis virus (RGNNV), a highly pathogenic RNA virus, is a major pathogen in cultured groupers, and causes huge economic losses. A series of host cellular proteins involved in RGNNV infection was identified. However, the impact of Rab1 on RGNNV infection has not yet been reported. In this study, a novel Rab1 homolog (EcRab1) from Epinephelus coioides was cloned, and its roles during virus infection and host immune responses were investigated. EcRab1 encoded a 202 amino acid polypeptide, showing 98% and 78% identity to Epinephelus lanceolatus and Homo sapiens, respectively. After challenge with RGNNV or poly(I:C), the transcription of EcRab1 was altered both in vitro and in vivo, implying that EcRab1 was involved in virus infection. Subcellular localization showed that EcRab1 was displayed as punctate structures in the cytoplasm, which was affected by EcRab1 mutants. The dominant negative (DN) EcRab1, enabling EcRab1 to remain in the GDP-binding state, caused EcRab1 to be diffusely distributed in the cytoplasm. Constitutively active (CA) EcRab1, enabling EcRab1 to remain in the GTP-binding state, induced larger cluster structures of EcRab1. During the late stage of RGNNV infection, some EcRab1 co-localized with RGNNV, and the size of EcRab1 clusters was enlarged. Importantly, overexpression of EcRab1 significantly inhibited RGNNV infection, and knockdown of EcRab1 promoted RGNNV infection. Furthermore, EcRab1 inhibited the entry of RGNNV to host cells. Compared with EcRab1, overexpression of DN EcRab1 or CA EcRab1 also promoted RGNNV infection, suggesting that EcRab1 regulated RGNNV infection, depending on the cycles of GTP- and GDP-binding states. In addition, EcRab1 positively regulated interferon (IFN) immune and inflammatory responses. Taken together, these results suggest that EcRab1 affects RGNNV infection, possibly by regulating host immunity. Our study furthers the understanding of Rab1 function during virus infection, thus helping to design new antiviral strategies.

How the luxR Gene Affects the Pathogenicity of Pseudomonas plecoglossicida and the Immune Response of Epinephelus coioides

This study aimed to investigate the effect of reduced expression of the luxR gene on the virulence of Pseudomonas plecoglossicida and the immune response of Epinephelus coioides. To achieve this, RNA interference (RNAi) was used to silence the luxR gene, and the pathogenicity of wild-type and luxR-RNAi strains of P. plecoglossicida, as well as the immune response of Epinephelus coioides to the infection of these two strains, were compared. The mutant strain with the highest silencing efficiency of 70.1% was selected for subsequent analysis. Silencing the luxR gene in the mutant strain resulted in a significant 30% reduction in mortality rates in artificially infected Epinephelus coioides compared to the wild-type strain. Transcriptome analysis revealed that the host transcriptome, particularly in the spleens of infected Epinephelus coioides, was markedly altered by the silencing of the luxR gene in the mutant strain. Tilapia infected with the luxR-RNAi strain exhibited altered immune defenses, with changes in gene expression primarily in the NOD-like receptor (NLR) signaling pathway. These results suggest that the luxR gene plays a crucial role in the host’s resistance to pathogen invasion, and reducing its expression could decrease quorum sensing (QS) signals while increasing the expression of the IL-1β gene in the host’s NLR pathway. This effect may lead to a pro-inflammatory response that enhances the immune response to infection. Further investigation of these mechanisms may lead to innovative approaches to treating bacterial infections.

Identification of circRNAs and circRNA-mRNA network of Epinephelus coioides during Singapore grouper iridovirus infection

Circular RNA (circRNA), one of the important non-coding RNA molecules with a closed-loop structure, plays a key regulatory role in cell processing. In this study, circRNAs of Epinephelus coioides, an important marine cultured fish in China, were isolated and characterized, and the network of circRNAs and mRNA was explored during Singapore grouper iridovirus (SGIV) infection, one of the most important double stranded DNA virus pathogens of marine fish. 10 g of raw data was obtained by high-throughput sequencing, and 2599 circRNAs were classified. During SGIV infection, 123 and 37 circRNAs occurred differential expression in spleen and spleen cells, indicating that circRNAs would be involved in the viral infection. GO annotation and KEGG demonstrated that circRNAs could target E. coioides genes to regulate cell activity and the activation of immune factors. The results provide some insights into the circRNAs mediated immune regulatory network during bony fish virus infection.

A fluorescence and colorimetric bifunctional probe for recognition of Al3+ and Cu2+ and its applications in cells and environmental water systems

A highly sensitive bifunctional probe (1) for Al3+ and Cu2+ by the reaction of 2-hydroxyl-1-naphthalene formaldehyde with indole-3-formyl hydrazine was synthesized. In THF-H2O (v/v = 3:7) solution, 1 only shows fluorescence enhanced recognition for Al3+ in 17 common metal ions, which shows that 1 can be used as a highly selective fluorescence enhancement probe for Al3+. When the concentration of Al3+ is 20.0 µM (1.0 equivalent), the fluorescence intensity of 1 is increased by 195 times, and the detection limit of 1 for Al3+ is calculated to be 3.8 nM, which shows that 1 is fluorescence probe for Al3+ with an ultra-high sensitivity. Trace Al3+ (For example, the WHO drinking water quality standard 7.4 μM) can be clearly observed by the fluorescence test paper covering 1 through the change of fluorescence color. In addition, the vivo-detection ability of 1 is verified by detected the Al3+ in living GS cells of epinephelus coioides. Furthermore, 1 can also be used as a high selective and sensitive colorimetric probe for Cu2+. In THF-H2O (v/v = 3:7) buffer solution (pH = 7.0), the addition of Cu2+ results in a 69 nm red shift in the UV–visible absorption spectrum of 1 solution, causing the color of 1 solution to change from colorless to yellow. In addition, 1 shows a low detection limit of 39.3 nM, a fast response time, excellent recyclability and anti-interference for Cu2+. Therefore, 1 can be used for fluorescence detection of Cu2+ in environmental water systems.