Comparative proteome analysis revealed potential biomarkers and the underlying immune mechanisms in Vibrio-resistant hybrid grouper, Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂.

Vibrio alginolyticus is the causative agent of vibriosis, a common bacterial infection in grouper aquaculture that is associated with the development of haemorrhagic and non-haemorrhagic ulcerations on the fish. In the present study, comparative proteome analysis was performed on serum samples from Vibrio-resistant and Vibrio-susceptible grouper. Samples were analysed using high-throughput LC-MS/MS and identified 2770 unique peptides that corresponded to 344 proteins. Subsequent analysis identified 21 proteins that were significantly up-regulated in the resistant group compared to the control and the susceptible groups. Those proteins are associated with immunostimulatory effects, signalling and binding cascade, metabolism, and maintaining tissue integrity and physiological condition. Besides, potential protein biomarkers related to the immune system were identified, which could be associated with the disease-resistant phenotype. These data provide insights into the underlying immune mechanism of hybrid groupers upon Vibrio sp. infection.

Selection and characterization of ssDNA aptamer targeting Macrobrachium rosenbergii nodavirus capsid protein: A potential capture agent in gold-nanoparticle-based aptasensor for viral protein detection.

The giant freshwater prawn holds a significant position as a valuable crustacean species cultivated in the aquaculture industry, particularly well-known and demanded among the Southeast Asian countries. Aquaculture production of this species has been impacted by Macrobrachium rosenbergii nodavirus (MrNV) infection, which particularly affects the larvae and post-larvae stages of the prawn. The infection has been recorded to cause mortality rates of up to 100% among the affected prawns. A simple, fast, and easy to deploy on-site detection or diagnostic method is crucial for early detection of MrNV to control the disease outbreak. In the present study, novel single-stranded DNA aptamers targeting the MrNV capsid protein were identified using the systematic evolution of ligands by exponential enrichment (SELEX) approach. The aptamer was then conjugated with the citrate-capped gold nanoparticles (AuNPs), and the sensitivity of this AuNP-based aptasensor for the detection of MrNV capsid protein was evaluated. Findings revealed that the aptamer candidate, APT-MrNV-CP-1 was enriched throughout the SELEX cycle 4, 9, and 12 with the sequence percentage of 1.76%, 9.09%, and 12.42%, respectively. The conjugation of APT-MrNV-CP-1 with citrate-capped AuNPs exhibited the highest sensitivity in detecting the MrNV capsid protein, where the presence of 62.5 nM of the viral capsid protein led to a significant agglomeration of the AuNPs. This study demonstrated the practicality of an AuNP-based aptasensor for disease diagnosis, particularly for detecting MrNV infection in giant freshwater prawns.

Riding the wave of innovation: immunoinformatics in fish disease control.

The spread of infectious illnesses has been a significant factor restricting aquaculture production. To maximise aquatic animal health, vaccination tactics are very successful and cost-efficient for protecting fish and aquaculture animals against many disease pathogens. However, due to the increasing number of immunological cases and their complexity, it is impossible to manage, analyse, visualise, and interpret such data without the assistance of advanced computational techniques. Hence, the use of immunoinformatics tools is crucial, as they not only facilitate the management of massive amounts of data but also greatly contribute to the creation of fresh hypotheses regarding immune responses. In recent years, advances in biotechnology and immunoinformatics have opened up new research avenues for generating novel vaccines and enhancing existing vaccinations against outbreaks of infectious illnesses, thereby reducing aquaculture losses. This review focuses on understanding in silico epitope-based vaccine design, the creation of multi-epitope vaccines, the molecular interaction of immunogenic vaccines, and the application of immunoinformatics in fish disease based on the frequency of their application and reliable results. It is believed that it can bridge the gap between experimental and computational approaches and reduce the need for experimental research, so that only wet laboratory testing integrated with in silico techniques may yield highly promising results and be useful for the development of vaccines for fish.

Oleic acid as potential immunostimulant in metabolism pathways of hybrid grouper fingerlings (Epinephelus fuscoguttatus × Epinephelus lanceolatus) infected with Vibrio vulnificus.

Grouper culture has been expanding in Malaysia due to the huge demand locally and globally. However, due to infectious diseases such as vibriosis, the fish mortality rate increased, which has affected the production of grouper. Therefore, this study focuses on the metabolic profiling of surviving infected grouper fed with different formulations of fatty acid diets that acted as immunostimulants for the fish to achieve desirable growth and health performance. After a six-week feeding trial and one-week post-bacterial challenge, the surviving infected grouper was sampled for GC-MS analysis. For metabolite extraction, a methanol/chloroform/water (2:2:1.8) extraction method was applied to the immune organs (spleen and liver) of surviving infected grouper. The distribution patterns of metabolites between experimental groups were then analyzed using a metabolomics platform. A total of 50 and 81 metabolites were putatively identified from the spleen and liver samples, respectively. Our further analysis identified glycine, serine, and threonine metabolism, and alanine, aspartate and glutamate metabolism had the most impacted pathways, respectively, in spleen and liver samples from surviving infected grouper. The metabolites that were highly abundant in the spleen found in these pathways were glycine (20.9%), l-threonine (1.0%) and l-serine (0.8%). Meanwhile, in the liver l-glutamine (1.8%) and aspartic acid (0.6%) were found to be highly abundant. Interestingly, among the fish diet groups, grouper fed with oleic acid diet produced more metabolites with a higher percent area compared to the control diets. The results obtained from this study elucidate the use of oleic acid as an immunostimulant in fish feed formulation affects more various immune-related metabolites than other formulated feed diets for vibriosis infected grouper.

A Comprehensive Review with Future Insights on the Processing and Safety of Fermented Fish and the Associated Changes.

As an easily spoiled source of valuable proteins and lipids, fish is preserved by fermentation in many cultures. Over time, diverse types of products have been produced from fish fermentation aside from whole fish, such as fermented fish paste and sauces. The consumption of fermented fish products has been shown to improve both physical and mental health due to the composition of the products. Fermented fish products can be dried prior to the fermentation process and include various additives to enhance the flavours and aid in fermentation. At the same time, the fermentation process and its conditions play a major role in determining the quality and safety of the product as the compositions change biochemically throughout fermentation. Additionally, the necessity of certain microorganisms and challenges in avoiding harmful microbes are reviewed to further optimise fermentation conditions in the future. Although several advanced technologies have emerged to produce better quality products and easier processes, the diversity of processes, ingredients, and products of fermented fish warrants further study, especially for the sake of the consumers' health and safety. In this review, the nutritional, microbial, and sensory characteristics of fermented fish are explored to better understand the health benefits along with the safety challenges introduced by fermented fish products. An exploratory approach of the published literature was conducted to achieve the purpose of this review using numerous books and online databases, including Google Scholar, Web of Science, Scopus, ScienceDirect, and PubMed Central, with the goal of obtaining, compiling, and reconstructing information on a variety of fundamental aspects of fish fermentation. This review explores significant information from all available library databases from 1950 to 2022. This review can assist food industries involved in fermented fish commercialization to efficiently ferment and produce better quality products by easing the fermentation process without risking the health and safety of consumers.

A Comprehensive Review on the Processing of Dried Fish and the Associated Chemical and Nutritional Changes.

Fish is a good source of nutrients, although it is easily spoiled. As such, drying is a common method of preserving fish to compensate for its perishability. Dried fish exists in different cultures with varying types of fish used and drying methods. These delicacies are not only consumed for their convenience and for their health benefits, as discussed in this review. Most commonly, salt and spices are added to dried fish to enhance the flavours and to decrease the water activity (aw) of the fish, which further aids the drying process. For fish to be dried effectively, the temperature, drying environment, and time need to be considered along with the butchering method used on the raw fish prior to drying. Considering the various contributing factors, several physicochemical and biochemical changes will certainly occur in the fish. In this review, the pH, water activity (aw), lipid oxidation, and colour changes in fish drying are discussed as well as the proximate composition of dried fish. With these characteristic changes in dried fish, the sensory, microbial and safety aspects of dried fish are also affected, revolving around the preferences of consumers and their health concerns, especially based on how drying is efficient in eliminating/reducing harmful microbes from the fish. Interestingly, several studies have focused on upscaling the efficiency of dried fish production to generate a safer line of dried fish products with less effort and time. An exploratory approach of the published literature was conducted to achieve the purpose of this review. This evaluation gathers important information from all available library databases from 1990 to 2022. In general, this review will benefit the fishery and food industry by enabling them to enhance the efficiency and safety of fish drying, hence minimising food waste without compromising the quality and nutritional values of dried fish.

Comparison of Different Dietary Fatty Acids Supplement on the Immune Response of Hybrid Grouper (Epinephelus fuscoguttatus × Epinephelus lanceolatus) Challenged with Vibrio vulnificus.

Aquaculture has been expanding in Malaysia due to the increased demand for fish products. In addition, aquaculture faces challenges in maintaining feed suitability in support of the global growth of fish production. Therefore, improvements in diet formulation are necessary to achieve the optimal requirements and attain a desirable growth efficiency and health performance in fish. Seven weeks of study were conducted to compare the equal amounts of different fatty acids (2%) (oleic acid, stearic acid, palmitic acid, and behenic acid) on the survival, the growth, and the immune response of hybrid grouper (Epinephelus fuscoguttatus × Epinephelus lanceolatus) against V. vulnificus. After six weeks of the feeding trial, fish were challenged with V. vulnificus for 30 min before continuing on the same feeding regime for the next seven days (post-bacterial challenge). Fish supplemented with dietary oleic acid showed significantly (p < 0.05) enhanced immune responses, i.e., lysozyme, respiratory burst, and phagocytic activities compared to the control diet group for both pre-and post-bacterial challenges. Following the Vibrio challenge, no significant effects of supplemented fatty acid diets on survival rate were observed, although dietary oleic acid demonstrated the highest 63.3% survival rate compared to only 43.3% of the control diet group. In addition, there were no significant effects (p > 0.05) on specific growth rate (SGR), white blood cell (WBC), and red blood cell (RBC) counts among all experimental diets. The results from this study suggest that among the tested dietary fatty acids, the oleic acid diet showed promising results in the form of elevated immune responses and increased disease resistance of the hybrid grouper fingerlings challenged with V. vulnificus.

LC-qTOF-MS analysis of fish immune organs reveals the distribution of amino acids in response to metabolic adaptation of the survival phenotype in grouper against Vibrio infection.

Epinephelus fuscoguttatus is economically crucial to various Southeast Asia countries where they are reared in fish farms to meet the demand for supply. However, a systemic infectious disease known as vibriosis has steadily and extensively affected the fish farming industry. The disease is caused by Vibrio spp., which are pathogenic gram-negative bacteria. This study focused on understanding the host's metabolic adaptation against Vibrio vulnificus infection, which features a survival phenotype, by profiling the metabolites in grouper fingerlings that survived the experimental infection. Mapping of the pathways is crucial to explain the roles of metabolites in fish immunity. A solvent extraction method was used on the grouper's immune organs (gills, liver and spleen) prior to Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry (LC-qTOF-MS) analysis. The metabolites identified in fingerlings that survived experimental infections were mostly amino acids (primary metabolites). Glutamine (0.44%), alanine (0.68%), phenylalanine (2.63%) and tyrosine (2.60%) were highly abundant in survived-infected gills. Aspartic acid (13.57%) and leucine (4.01%) were highly abundant in the livers of the survived-infected fish and lysine was highly abundant in both gills (2.94%) and liver (3.64%) of the survived-infected fish. Subsequent bioinformatics analysis revealed the involvement of the identified functional amino acids in various immune-related pathways. The current findings facilitate the comprehension of the metabolic adaptation of grouper fingerlings that exhibited a survival phenotype against Vibrio infection. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03269-1.

Omics Strategies in Current Advancements of Infectious Fish Disease Management.

Aquaculture is an important industry globally as it remains one of the significant alternatives of animal protein source supplies for humankind. Yet, the progression of this industry is being dampened by the increasing rate of fish mortality, mainly the outbreak of infectious diseases. Consequently, the regress in aquaculture ultimately results in the economy of multiple countries being affected due to the decline of product yields and marketability. By 2025, aquaculture is expected to contribute approximately 57% of fish consumption worldwide. Without a strategic approach to curb infectious diseases, the increasing demands of the aquaculture industry may not be sustainable and hence contributing to the over-fishing of wild fish. Recently, a new holistic approach that utilizes multi-omics platforms including transcriptomics, proteomics, and metabolomics is unraveling the intricate molecular mechanisms of host-pathogen interaction. This approach aims to provide a better understanding of how to improve the resistance of host species. However, no comprehensive review has been published on multi-omics strategies in deciphering fish disease etiology and molecular regulation. Most publications have only covered particular omics and no constructive reviews on various omics findings across fish species, particularly on their immune systems, have been described elsewhere. Our previous publication reviewed the integration of omics application for understanding the mechanism of fish immune response due to microbial infection. Hence, this review provides a thorough compilation of current advancements in omics strategies for fish disease management in the aquaculture industry. The discovery of biomarkers in various fish diseases and their potential advancement to complement the recent progress in combatting fish disease is also discussed in this review.

Gut Immune System and the Implications of Oral-Administered Immunoprophylaxis in Finfish Aquaculture.

The gastrointestinal immune system plays an important role in immune homeostasis regulation. It regulates the symbiotic host-microbiome interactions by training and developing the host's innate and adaptive immunity. This interaction plays a vital role in host defence mechanisms and at the same time, balancing the endogenous perturbations of the host immune homeostasis. The fish gastrointestinal immune system is armed with intricate diffused gut-associated lymphoid tissues (GALTs) that establish tolerance toward the enormous commensal gut microbiome while preserving immune responses against the intrusion of enteric pathogens. A comprehensive understanding of the intestinal immune system is a prerequisite for developing an oral vaccine and immunostimulants in aquaculture, particularly in cultured fish species. In this review, we outline the remarkable features of gut immunity and the essential components of gut-associated lymphoid tissue. The mechanistic principles underlying the antigen absorption and uptake through the intestinal epithelial, and the subsequent immune activation through a series of molecular events are reviewed. The emphasis is on the significance of gut immunity in oral administration of immunoprophylactics, and the different potential adjuvants that circumvent intestinal immune tolerance. Comprehension of the intestinal immune system is pivotal for developing effective fish vaccines that can be delivered orally, which is less labour-intensive and could improve fish health and facilitate disease management in the aquaculture industry.

Peculiarities of innate immune memory in crustaceans.

Classical characteristic of the innate immune system is the lack of ability to build up immunological memory, contrast to the adaptive immune system that is capable of "remembering" antigens, and rapidly mount a greater magnitude of immune response upon subsequent exposure to the same antigens. Peculiarly, immunological memory of innate immunity is evidenced in invertebrates. At least three different memory phenomena have been described, namely sustained unique response, recalled response, and immune shift. Studies attended to decipher the mechanistic biology of the innate immune memory reveals the role of epigenetics, which modulates the response of immune memory, and the heritability of immune memory to subsequent generations. A parthenogenetic Artemia model demonstrated successful transgenerational epigenetic inheritance of resistance trait against Vibrio campbellii. Following, the role of invertebrate hemocytes and Down syndrome cell adhesion molecule (Dscam) in innate immune memory is reviewed. While there is no vertebrate antibody homolog found in invertebrates, Dscam was found to resemble the functionality of vertebrate antibody. Insight of Dscam as immune factor was illustrated further in the current review.

Comparative genome analysis reveals a distinct influence of nucleotide composition on virus-host species-specific interaction of prawn-infecting nodavirus.

Discovery of species-specific interaction between the host and virus has drawn the interest of many researchers to study the evolution of the newly emerged virus. Comparative genome analysis provides insights of the virus functional genome evolution and the underlying mechanisms of virus-host interactions. The analysis of nucleotide composition signified the evolution of nodavirus towards host specialization in a host-specific mutation manner. GC-rich genome of betanodavirus was significantly deficient in UpA and UpU dinucleotides composition, whilst the AU-rich genome of gammanodavirus was deficient in CpG dinucleotide. The capsid of MrNV and PvNV of gammanodavirus retains the highest abundance of adenine and uracil at the second codon position, respectively, which were found to be very distinctive from the other genera. ENC-GC3 plot inferred the influence of natural selection and mutational pressure in shaping the evolution of MrNV RdRp and capsid, respectively. Furthermore, CAI/eCAI analysis predicts a comparable adaptability of MrNV in squid, Sepia officinalis than its natural host, Macrobrachium rosenbergii. Thus, further study is warranted to investigate the capacity of MrNV replication in S. officinalis owing to its high codon adaptation index.

Evaluation of potential molecular interaction between quorum sensing receptor, LuxP and grouper fatty acids: in-silico screening and simulation.

Pathologically relevant behaviors of Vibrio, such as the expression of virulence factors, biofilm production, and swarming motility, have been shown to be controlled by quorum sensing. The autoinducer-2 quorum sensing receptor protein LuxP is one of the target proteins for drug development to suppress the virulence of Vibrio. Here, we reported the potential molecular interaction of fatty acids identified in vibriosis-resistant grouper with LuxP. Fatty acid, 4-oxodocosahexaenoic acid (4R8) showed significant binding affinity toward LuxP (-6.0 kcal/mol) based on molecular docking analysis. The dynamic behavior of the protein-ligand complex was illustrated by molecular dynamic simulations. The fluctuation of the protein backbone, the stability of ligand binding, and hydrogen bond interactions were assessed, suggesting 4R8 possesses potential interaction with LuxP, which was supported by the low binding free energy (-29.144 kJ/mol) calculated using the molecular mechanics Poisson-Boltzmann surface area.

Molecular biology of Macrobrachium rosenbergii nodavirus infection in giant freshwater prawn.

Macrobrachium rosenbergii nodavirus (MrNV) has been threatening the giant freshwater prawn aquaculture since 1997, causing white tail disease in the prawn species that leads to 100% lethality of the infected postlarvae. Comprehension of the viral infectivity and pathogenesis at molecular biology level has recently resolved the viral capsid protein and evidenced the significant difference in the viral structural protein compared to other nodaviruses that infect fish and insect. Cumulative researches have remarked the proposal to assert MrNV as a member of new genus, gammanodavirus to the Nodaviridae family. The significance of molecular biology in MrNV infection is being highlighted in this current review, revolving the viral life cycle from virus binding and entry into host, virus replication in host cell, to virus assembly and release. The current review also highlights the emerging aptamers technology that is also known as synthetic antibody, its application in disease diagnosis, and its prophylactic and therapeutic properties. The future perspective of synthetic virology technology in understanding viral pathogenesis, as well as its potential in viral vaccine development, is also discussed.

Protective effect of apple mangrove Sonneratia caseolaris extract in Edwardsiella tarda-infected African catfish, Clarias gariepinus.

Outbreaks of edwardsiellosis have severe impact on the aquaculture production of African catfish Clarias gariepinus. In this study, feed supplemented with apple mangrove Sonneratia caseolaris extract was evaluated for its protective effect against Edwardsiella tarda infection in African catfish. Results showed an increase in growth performance and higher survival rate in the treatment groups in a dose dependent manner. Haematological analyses showed an increase in white blood cell count in the treatment groups. Histopathological analysis revealed degenerative changes and regeneration of liver tissue architecture in both the control and treatment groups. However, the presence of inflammatory cells was found exclusively in the kidney of T3 treatment group that was supplemented with the highest dose of extract at 3.17 mg/ml, which inferred the activation of immune response in the fish. Contrast to the deteriorative alteration observed in the kidney of the control group due to E. tarda infection, treatment group exhibited tissue regeneration and well-defined kidney tissue architecture at 3 dpi. Taken together, these results demonstrated that supplementation with the methanol extract of S. caseolaris possesses protective effect in African catfish against the infection of E. tarda.

Transcriptome data of Epinephelus fuscoguttatus infected by Vibrio vulnificus.

Vibriosis disease by Vibrio spp. greatly reduced productivity of aquaculture, such as brown-marbled grouper (Epinephelus fuscoguttatus), which is an economically important fish species in Malaysia. Preventive measures and immediate treatment are critical to reduce the mortality of E. fuscoguttatus from vibriosis. To investigate the molecular mechanisms associated with immune response and host-bacteria interaction, a transcriptomic analysis was performed to compare between healthy and Vibrio-infected groupers. This permits the discovery of immune-related genes, specifically the resistance genes upon infection. Herein, we provide the raw transcriptome data from Illumina HiSeq. 4000 that have been deposited into NCBI SRA database with the BioProject accession number PRJNA396437. A total of 493,403,076 raw sequences of 74.5 Gb were obtained. Trimming of the raw data produced 437,186,232 clean reads of ~58 Gb. These datasets will be useful to elucidate the defence mechanisms of E. fuscoguttatus against Vibrio vulnificus infection for future development of effective prevention and treatment of vibriosis.

Data set of phylogenetic analysis inferred based on the complete genomes of the family Nodaviridae.

In this article, nine complete genomes of viruses from the genus Alphanodavirus and Betanodavirus (Family Nodaviridae) were comparatively analyzed and the data of their evolutionary origins and relatedness are reported. The nucleotide sequence alignment of the complete genomes from all species and their deduced evolutionary relationships are presented. High sequence similarity within the genus Betanodavirus compared to the genus Alphanodavirus was revealed in multiple sequence alignment of the Nodaviridae genomes. The amino acid sequence similarity for both RNA1 and RNA2 ORF is more conserved in Betanodavirus, compared to Alphanodavirus. The conserved and variable regions within the virus genome that were defined based on the multiple sequence alignments are presented in this dataset.

The antioxidant effect of Beackea frutescence microemulsions dietary supplements on skin absorption studies.

Objective: To study the effect of two different microemulsions containing Beackea frutescence supplements composed of nerolidool, selenium and vitamin E on absorption effect related to skin health and skin aging. Materials and methods: A total of 39 volunteers with normal and healthy skin were divided into three groups (n = 13) and supplemented for a period of 12 weeks. Group 1 received a mixture of lutein (3 mg/day), lycopene (3 mg/day), α-tocopherol (10 mg/day), selenium (75 µg/day) and ß-nerolidool (4.8 mg/day) and Group 2 was supplemented with a mixture of ß-nerolidool (4.8 mg/day), lycopene (6 mg/day), selenium (75 µg/day) and α-tocopherol (10 mg/day). Group 3 was the placebo control. Wrinkling, smoothness, scaling and roughness of the skin were determined by Surface Evaluation of Living Skin (Visioscan). Results: Upon supplementation, serum levels of selected nerolidool increased in both groups. Skin thickness and density were determined by ultrasound measurements. A significant increase for both parameters was determined in the serum groups. Roughness and scaling were improved by the supplementation with antioxidant micronutrients. In the placebo group, no changes were found for any of the parameters. Conclusion:Beackeafrutescence microemulsion supplements have shown significant change in the texture of human skin as well as scaling, wrinkling, smoothness and roughness were improved by the supplementation.

Adiponectin SNP45TG is associated with gestational diabetes mellitus.

INTRODUCTION: Diabetes and pregnancy can be associated in two ways: pregnancy that occurs in women who are already diabetic (diabetes of pre-gestational origin); and diabetes that occur in women who are already pregnant [gestational diabetes mellitus (GDM) (O'sullivan 1961)]. Patients with previous GDM history have higher risk of developing diabetes outside of pregnancy. Accumulating literature had suggested that adiponectin plays a role in the pathophysiology of this metabolic syndrome, and several of the common single nucleotide polymorphisms (SNP) in adiponectin gene have been identified in type 2 diabetes. Thus, one of the commonly found SNP was studied to determine its association with GDM. OBJECTIVE: To identify the association of SNP45TG with GDM. METHODS: This is a cross-sectional study involving pregnant mothers of <18 gestational weeks, who were recruited from three local antenatal clinics in Selangor, Malaysia. Their genomic DNA was extracted from EDTA treated whole blood using commercialized kit. Adiponectin gene was amplified through conventional PCR and SNP was detected using restriction enzyme SmaI. Plasma adiponectin level, fructosamine level and HbA(1c) percentage were also examined. RESULTS: Among the 79 antenatal patients recruited, 53 patients were normal and 26 were diagnosed with GDM. Among the 53 normal patients, 18 carry TG/GG genotype. Meanwhile, among the 26 patients that were diagnosed with GDM 15 carry TG/GG genotype. Significant association was found between SNP45TG with GDM ( χ(2) = 4.038; P < 0.05). In addition, normal patients with TT genotype have significantly higher plasma adiponectin level compared to other groups. CONCLUSION: We concluded that SNP45TG in adiponectin gene is associated with the occurrence of GDM.