Stability characterizations of feed-based bivalent vaccine containing inactivated Streptococcus agalactiae and Aeromonas hydrophila against streptococcosis and Aeromonas infections in red hybrid tilapia (Oreochromis sp.).

Feed-based bivalent vaccine (FBBV) containing killed whole organism (KWO) of Streptococcus agalactiae and Aeromonas hydrophila with 10% palm oil was previously proved to improve red hybrid tilapia's (Oreochromis sp.) immunity against streptococcosis and Aeromonas infections. This study characterized the FBBV's stability following the preparatory process and storage. The FBBV was prepared, and the KWO's stability was determined microscopically and molecularly. The efficacy of FBBV stored at room temperature (25 ± 2 °C) for 0, 30 and 60 days was investigated in red hybrid tilapia. The results indicated the addition of palm oil was not affecting the KWO's structure and helping in the FBBV's pelletization. In 1 g of FBBV contained 1.5 × 109 CFU/g of S. agalactiae and 4.9 × 109 CFU/g of A. hydrophila, respectively, even after 60 days of storage at room temperature. The KWO's structure in FBBV was not affected following in vitro acidic tolerance analysis, as noted from light and electron microscopies. The FBBV's carbohydrate, energy, moisture, total protein and total ash contents remained stable at 95% after 60 days of storage at room temperature, while the KWO's concentration was slightly reduced to 83.3% for S. agalactiae (1.25 × 109 CFU/g) and 80.6% for A. hydrophila (3.85 × 109 CFU/g), respectively. Fish vaccinated with FBBV that was stored for 0, 30 and 60 days did not show any significant differences (p ≥ 0.05) in the relative percent survival when challenged with pathogenic Streptococcus spp. and Aeromonas spp. These findings suggested that the FBBV is a stable vaccine, which underscores its potential application as aquatic vaccines in aquaculture.

Streamlined boiling lysis DNA extraction for Gram-positive aquaculture pathogen Streptococcus agalactiae.

Accurate genetic analysis is essential for the detection of pathogens as it necessitates suitable DNA extraction methods tailored to specific microorganisms such as Gram-positive bacteria. This study examined several commercial and simplified DNA extraction methods for their suitability in isothermal downstream applications. Extracted DNA was assessed using spectrophotometry, electrophoresis, polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) while its stability was inspected after five months of storage. The findings revealed variations in DNA yield, purity and integrity among the extraction methods. While extraction kits demonstrated high yield and purity, the in-house extraction techniques showed incoherent correlation between yield and purity, yet showed promise for a streamlined extraction process. The DNA acquired from all methods yielded positive amplification in PCR and LAMP. DNA extracted by kits exhibits prolonged stability than those obtained via boiling lysis. Both methods offer distinct advantages, with commercial kits providing longer stability and high-quality DNA while boiling lysis stands out for its simplicity, with shorter handling and processing periods. This study emphasizes selecting ideal extraction methods for Streptococcus agalactiae, in the prospect of aquaculture settings. In particular, successful LAMP reaction suggests that boiled extracts are feasible enough for detection, and suited for point-of-care (POC) testing where prompt detection of aquatic pathogens is often critical. Ultimately, the choice of method should be contemplated on a case-by-case basis such as the study goals, intended settings, and type of samples.

Expression of immuno-transcriptome response in red hybrid tilapia (Oreochromis sp.) hindgut following vaccination with feed-based bivalent vaccine.

Streptococcosis and aeromoniasis are the main obstacles to sustainable tilapia production. Vaccination offered an effective method to control microbial infections. Previously, a feed-based bivalent vaccine (FBBV) containing killed whole organisms of Streptococcus agalactiae and Aeromonas hydrophila mixed with 10% palm oil was successfully developed, which provided good protection against streptococcosis and aeromoniasis in Oreochromis sp. However, the mechanisms of immunities in vaccinated fish still need clarification. Here, the hindgut transcriptome of vaccinated and control fish was determined, as the gut displays higher affinity towards antigen uptake and nutrient absorption. The efficacy of FBBV to improve fish immunity was evaluated according to the expression of immune-related genes in the vaccinated fish hindgut throughout the 8-week experimental period using RT-qPCR. The vaccinated fish hindgut at week 6 was further subjected to transcriptomic analysis due to the high expression of immune-related genes and contained killed whole organisms. Results demonstrated the expression of immune-related genes was in correlation with the presence of killed whole organisms in the vaccinated fish hindgut. Transcriptomic analysis has allowed the prediction of robust immune-related pathways, including innate and adaptive immunological responses in vaccinated fish hindgut than control fish. Pathways related to the regulation of lipid metabolism and modulation of the immune system were also significantly enriched (p ≤ .05). Overall, results offer a fundamental study on understanding the immunological response in Oreochromis sp. following vaccination with the FBBV pellet and support further application to prevent bacterial diseases in aquaculture.

Intraperitoneal Injection of the Porphyromonas gingivalis Outer Membrane Vesicle (OMV) Stimulated Expressions of Neuroinflammatory Markers and Histopathological Changes in the Brains of Adult Zebrafish.

Porphyromonas gingivalis is the major pathogenic bacteria found in the subgingival plaque of patients with periodontitis, which leads to neuroinflammation. The bacteria destroy periodontal tissue through virulence factors, which are retained in the bacteria's outer membrane vesicles (OMV). This study aimed to determine the real-time effect of an intraperitoneal injection of P. gingivalis OMV on the production and expression of inflammatory markers and histopathological changes in adult zebrafishes' central nervous systems (CNS). Following the LD50 (21 µg of OMV), the zebrafish were injected intraperitoneally with 18 µg of OMVs, and the control group were injected with normal saline at seven different time points. Brains of experimental zebrafish were dissected at desired time points for colorimetric assays, ELISA, and histology. This study discovered that nitric oxide and PGE2 were significantly increased at 45 min, while IL-1ß and IL-6 were expressed at subsequent 12 h and 24 h time points, respectively. Histopathological changes such as blood coagulation, astrocytosis, edema, spongiosis, and necrosis were observed between the 6hour and 24 h time points. The two apoptotic enzymes, caspases 3 and 9, were not expressed at any point. In summary, the OMV-induced neuroinflammatory responses and histopathological changes in adult zebrafish were time-point dependent. This study will enrich our understanding of the mechanism of P. gingivalis OMVs in neuroinflammation in a zebrafish model, most especially the timing of the expression of inflammatory mediators in relation to observable changes in brain tissues.

Natural Concurrent Infection of Vibrio harveyi and V. alginolyticus in Cultured Hybrid Groupers in Malaysia.

In September 2016, a marine fish farm operator in Selangor, Malaysia, reported a disease outbreak affecting juvenile hybrid groupers (Camouflage Grouper Epinephelus polyphekadion × Tiger Grouper E. fuscoguttatus). The average daily mortality was 120 fish, resulting in a cumulative mortality rate of 29% within 10 d. The affected hybrid groupers displayed lethargy, excessive mucus production, rotten fins, congestion of livers and kidneys, and enlargement of spleens. Microscopically, general congestion of the brains and internal organs was evident. Vibrio harveyi and V. alginolyticus were successfully isolated from the diseased fish. The isolated pathogens were found to be sensitive to oxytetracycline and tetracycline, but resistant towards ampicillin and vancomycin. Experimental infections using the isolated V. harveyi (108  CFU/mL), V. alginolyticus (108  CFU/mL), and concurrent infection by V. harveyi (108  CFU/mL) and V. alginolyticus (108  CFU/mL) in juvenile Asian Seabass Lates calcarifer resulted in 60, 100, and 100% mortality, respectively, within 240 h postinfection. The experimentally infected Asian Seabass demonstrated similar clinical signs and histopathological changes as the naturally infected hybrid groupers. However, concurrently infected fish demonstrated severe clinical signs and histopathological changes compared with single infections. These results suggest that both isolates of Vibrio are pathogenic to fish and responsible for the disease outbreak. However, concurrent infection involving V. alginolyticus and V. harveyi leads to a more devastating impact to the cultured fish. This is the first report of concurrent Vibrio infection in cultured marine fish in Malaysia.

Histological features of the gastrointestinal tract of wild Indonesian shortfin eel, Anguilla bicolor bicolor (McClelland, 1844), captured in Peninsular Malaysia.

This study was conducted to record the histological features of the gastrointestinal tract of wild Indonesian shortfin eel, Anguilla bicolor bicolor (McClelland, 1844), captured in Peninsular Malaysia. The gastrointestinal tract was segmented into the oesophagus, stomach, and intestine. Then, the oesophagus was divided into five (first to fifth), the stomach into two (cardiac and pyloric), and the intestine into four segments (anterior, intermediate, posterior, and rectum) for histological examinations. The stomach had significantly taller villi and thicker inner circular muscles compared to the intestine and oesophagus. The lamina propria was thickest in stomach, significantly when compared with oesophagus, but not with the intestine. However, the intestine showed significantly thicker outer longitudinal muscle while gastric glands were observed only in the stomach. The histological features were closely associated with the functions of the different segments of the gastrointestinal tract. In conclusion, the histological features of the gastrointestinal tract of A. b. bicolor are consistent with the feeding habit of a carnivorous fish.

Red hybrid tilapia (Oreochromis sp.) hindgut RNA-seq data after oral vaccination with a feed-based bivalent vaccine.

Previous studies have proven that red hybrid tilapia (Oreochromis sp.) vaccinated with a feed-based bivalent vaccine incorporating the formalin-killed whole organisms Streptococcus agalactiae and Aeromonas hydrophila mixed with 10 % palm oil showed good protection against streptococcosis and aeromoniasis diseases. However, the molecular mechanisms related to the induction of fish's immunological responses after vaccination are poorly investigated. Therefore, a transcriptomic study using the hindgut of red hybrid tilapia after vaccination was conducted, as the gut plays a role in antigen uptake and nutrient absorption. The transcriptome dataset has the potential to provide an understanding of the early induction of immunological responses in red hybrid tilapia after vaccination. Here, the vaccinated and control red hybrid tilapia's hindgut ribonucleic acid sequencing (RNA-seq) dataset was presented, which are available in the National Center for Biotechnology Information (NCBI) database with accession number PRJNA1014699.

Genome Characterization and Phylogenetic Analysis of Scale Drop Disease Virus Isolated from Asian Seabass (Lates calcarifer).

Scale drop disease virus (SDDV), a double-stranded DNA virus in the family Iridoviridae, has been reported widely in southeast Asian countries as a causative agent of scale drop syndrome (SDS) in Asian seabass. SDS has resulted in high mortality and significant economic losses to the aquaculture industry. This study demonstrated the use of metagenomic methods to investigate bacterial and viral communities present in infected fish tissues and recover a complete genome of the causative agent named SDDV TH7_2019. Characterization of the TH7_2019 genome revealed a genome size of 131 kb with 134 putative ORFs encoding viral proteins potentially associated with host apoptosis manipulation. A comparative genome analysis showed a high degree of amino acid identity across SDDV strains, with variations in number of repeat sequences and mutations within core genes. Phylogenetic analyses indicate a close relationship among SDDV genomes. This research enhances our understanding of the genetic diversity and evolutionary relationship of SDDV, contributing valuable insights for further development of effective control strategies of SDDV.

Evaluating the Intestinal Immunity of Asian Seabass (Lates calcarifer, Bloch 1790) following Field Vaccination Using a Feed-Based Oral Vaccine.

This study describes the levels of gut lysozyme and IgM, the number, size and density of gut-associated lymphoid tissue (GALT) regions, and the lymphocyte population in Asian seabass following field oral administration of a feed-based vaccine. Fish in a grow-out farm were selected and divided into two groups; Group 1 was vaccinated at week 0, 2, and 6, while Group 2 was not vaccinated. Samplings were done at 2-week intervals when the fish were observed for clinical signs, and gross lesions were recorded. The intestinal tissue and gut lavage fluid were collected. GALT regions (numbers, size, density and population of lymphocytes) were analyzed. Clinical signs such as abnormal swimming pattern and death, and gross lesions including scale loss, ocular opacity, and skin ulceration were observed in both groups. At the end of the study, the incidence rate between both groups were significantly different (p < 0.05). The gut IgM level and lysozyme activity, lymphocyte population, number, size and density of GALT regions of Group 1 were significantly (p < 0.05) higher than Group 2. Therefore, this study concludes that the feed-based vaccine reduces the incidence of vibriosis by stimulating the gut immunity of the vaccinated fish with an enhanced GALT region, specific IgM production against Vibrio harveyi, and lysozyme responses.

Distribution, Prevalence, and Antibiotic Susceptibility Profiles of Infectious Noncholera Vibrio Species in Malaysia.

Background: The noncholera Vibrio spp. which cause vibriosis are abundantly found in our water ecosystem. These bacteria could negatively affect both humans and animals. To date, there is a paucity of information available on the existence and pathogenicity of this particular noncholera Vibrio spp. in Malaysia in comparison to their counterpart, Vibrio cholera. Methods: In this study, we extracted retrospective data from Malaysian surveillance database. Analysis was carried out using WHONET software focusing noncholera Vibrio spp. including Vibrio parahaemolyticus, Vibrio vulnificus, Vibrio fluvialis, Vibrio alginolyticus, Vibrio hollisae (Grimontia hollisae), Vibrio mimicus, Vibrio metschnikovii, and Vibrio furnissii. Results: Here, we report the first distribution and prevalence of these species isolated in Malaysia together with the antibiotic sensitivity profile based on the species. We found that V. parahaemolyticus is the predominant species isolated in Malaysia. Noticeably, across the study period, V. fluvialis is becoming more prevalent, as compared to V. parahaemolyticus. In addition, this study also reports the first isolation of pathogenic V. furnissii from stool in Malaysia. Conclusion: These data represent an important step toward understanding the potential emergence of noncholera Vibrio spp. outbreaks.

Fish-associated Streptococcus agalactiae ST283: first human cases reported from Malaysia.

In South East Asia, Streptococcus agalactiae ST283 causes sepsis in healthy adults. Raw freshwater fish consumption is the only known risk factor. These two case reports are the first from Malaysia. Although they cluster with Singapore ST283, the epidemiology is complicated by the flow of people and fish across borders.

Immunogenicity and Efficacy of a Feed-Based Bivalent Vaccine against Streptococcosis and Motile Aeromonad Septicemia in Red Hybrid Tilapia (Oreochromis sp.).

Streptococcosis and motile Aeromonad septicemia (MAS) are the main bacterial diseases in tilapia culture worldwide, causing significant economic losses. Vaccination is an effective method of preventing diseases and contributes to economic sustainability. This study investigated the immuno-protective efficacy of a newly developed feed-based bivalent vaccine against streptococcosis and MAS in red hybrid tilapia. The feed-based bivalent vaccine pellet was developed by incorporating the formalin-killed S. agalactiae and A. hydrophila antigens into a commercial feed pellet with palm oil as the adjuvant. The bivalent vaccine was subjected to feed quality analyses. For immunological analyses, 900 fish (12.94 ± 0.46 g) were divided into two treatment groups in triplicate. Fish in Group 1 were unvaccinated (control), while those in Group 2 were vaccinated with the bivalent vaccine. The bivalent vaccine was delivered orally at 5% of the fish's body weight for three consecutive days on week 0, followed by boosters on weeks 2 and 6. Lysozyme and enzyme-linked immunosorbent assays (ELISAs) on serum, gut lavage, and skin mucus were performed every week for 16 weeks. Lysozyme activity in vaccinated fish was significantly (p ≤ 0.05) higher than in unvaccinated fish following vaccination. Similarly, the IgM antibody levels of vaccinated fish were significantly (p ≤ 0.05) higher after vaccination. The bivalent vaccine provided high protective efficacy against S. agalactiae (80.00 ± 10.00%) and A. hydrophila (90.00 ± 10.00%) and partial cross-protective efficacy against S. iniae (63.33 ± 5.77%) and A. veronii (60.00 ± 10.00%). During the challenge test, fewer clinical and gross lesions were observed in vaccinated fish compared with unvaccinated fish. Histopathological assessment showed less severe pathological changes in selected organs than the unvaccinated fish. This study showed that vaccination with a feed-based bivalent vaccine improves immunological responses in red hybrid tilapia, and thus protects against streptococcosis and MAS.

Phenotypic and genotypic comparison of pathogenic group B Streptococcus isolated from human and cultured tilapia (Oreochromis species) in Malaysia.

Group B Streptococcus (GBS) is a major cause of several infectious diseases in humans and fish. This study was conducted to compare human and fish-derived GBS in terms of their antimicrobial susceptibility, serotype, virulence and pili genes and sequence type (ST), and to determine whether there is a potential linkage of zoonotic transmission in Malaysia. GBS isolated from humans and fish had similar phenotypic characteristics and differed in virulence gene profile, antimicrobial susceptibility, serotype and sequence type. Fish GBS isolates had lower genetic diversity and higher antibiotic susceptibility than human isolates. We report a rare detection of the potentially fish-adapted ST283 in human GBS isolates. Both human and fish ST283 shared several phenotypic and genotypic features, including virulence and pilus genes and antimicrobial susceptibility, illustrating the value of monitoring GBS within the One Health scope. In this study, two human GBS ST283 isolates belonging to the variant common in fish hosts were identified, raising awareness of the zoonotic potential between the different species in Malaysia.

Toxicity and teratogenicity effects of valproic acid on zebrafish (Danio rerio) embryos in relation to autism spectrum disorder.

Valproic acid (VPA) is a widely prescribed antiepileptic drug with various medicinal efficacies. Accumulated evidence implied that prenatal exposure to VPA is highly associated with autism spectrum disorder (ASD). In this study, the zebrafish were exposed to a set of VPA concentrations (0, 5, 10, 20, 40, 80, 160, 320, 640, 1280, and 2560 µM) at 5 h post fertilization (hpf) to 120 hpf. The adverse effects of VPA were extensively studied through the evaluations on the mortality, heartbeats, spontaneous tail coiling, and hatching rate. Morphological observations were conducted at 120 hpf, following the exposure termination. Basic locomotor responses and anxiety-like behavioral alterations evaluated for behavioral impairments are the hallmark feature of ASD. The exposure to VPA at teratogenic concentrations reduced the aforementioned parameters in a dose-dependent manner (p ≤ .05). At the selected non-teratogenic concentrations of VPA, the treated larvae demonstrated profound alterations of basic locomotor responses. No significant changes of anxiety and thigmotactic behaviors were observed on the VPA-treated fish compared to the control (p ≥ .005). This study depicted that embryonic zebrafish exposure to VPA produced significant toxicity and teratogenicity effects as well as the alterations of basic behavioral responses. Overall, this study provides a fundamental insight of the toxicity effects at morphological and behavioral levels to facilitate the understanding of ASD mechanisms at different molecular levels.

Polystyrene microplastics induce gut microbiome and metabolome changes in Javanese medaka fish (Oryzias javanicus Bleeker, 1854).

Microplastics (MPs) have become emerging pollutants of public health concern, due to their impact on aqua-terrestrial ecosystems and integration into the food web, with evidence of human exposure and unrevealed health implications. There is a paucity of information regarding the effects of MPs exposure on the gut system using metagenomic and metabolomic approaches. In this study, Javanese medaka fish was exposed to 5 µm beads of polystyrene microplastics (PS-MPs) suspensions, at concentrations of 100 µg/L (MP-LOW), 500 µg/L (MP-MED), and 1000 µg/L (MP-HIGH), for a duration of 21 days, and evaluated for gut microbiome and metabolome responses. The results revealed a significant reduction (p < 0.05) in richness and diversity of the gut microbiome in the MP-HIGH group, and identification of 7 bacterial genera as differential features by the Linear discriminant analysis Effect Size (LEfSe). The gut metabolic profile revealed upregulation of 9 metabolites related to energy metabolism, via tricarboxylic acid cycle (TCA), creatine pathway, and urea cycle, as determined by the pathway analysis. Furthermore, positive correlation was found between the genus Aeromonas and glucose, lactate, and creatine metabolites. The study revealed that PS-MPs exposure resulted in altered bacterial microbiome and metabolic disorder related to energy metabolism. It further provided additional data on gut bacterial genera and metabolites associated with MPs toxicity in aquatic organism, which will inevitably enable its future health risks assessment in animals and possibly humans.

The Burden of Microplastics Pollution and Contending Policies and Regulations.

The relationship between humans and plastics has become intricate due to their versatile nature and low production cost. Plastics generation has surpassed that of other manufactured products, which, coupled with the prevailing poor waste management systems, makes it a serious problem for the terrestrial and aquatic environments as its final destination. Their extensive presence has continued to pose a significant threat, not only to the aquatic ecosystem but also to the approximately 3 billion people relying on it for their livelihood. Even more disturbing were the recent findings of these plastics in food and drinking water and the evidence of human exposure, the long-term health effects of which are largely unknown. This ubiquitous phenomenon has over time put plastics under critical observation, leading to the development of many local and international policies, resolutions, and directives aimed at addressing and reversing the menace. This review provided the first snapshot of the global and local governance strategies currently aimed at mitigating plastic pollution, their limitations, and future directions. The findings of the review revealed several aspects of microplastics (MPs) pollution to be overlooked in policy formulation, a laxity in policy implementation, and an apparent lack of indices to ascertain the impact of the regulations. Furthermore, there is currently no regulation on MPs contamination of food and drinking water and an apparent lack of funding for research into the health effects of plastics and their alternatives. This, therefore, necessitates the need for a well-coordinated approach at international and national levels to scale up these policies in all countries and translate them from paper to measurable, holistic, and realizable actions that will address all forms of plastic pollution.

Toxicity of Zinc Oxide Nanoparticles on the Embryo of Javanese Medaka (Oryzias javanicus Bleeker, 1854): A Comparative Study.

(1) Background: Zinc oxide nanoparticles (ZnO NPs) are widely applied in various human products. However, they can be extremely toxic for aquatic organisms, particularly fish. This research was conducted to determine the LC50 of ZnO NPs on the embryos of Javanese medaka (Oryzias javanicus) in ultra-pure, deionized, and dechlorinated tap water; (2) Methods: The experiments were conducted in a completely randomized design (CRD) with three replicates for six treatments for acute (0.100, 0.250, 0.500, 1.00, 5.00, and 10.00 mg/L) exposures for each type of water; (3) Results: The LC50 of ZnO NPs at 96 h was determined as 0.643 mg/L in ultra-pure water, 1.333 mg/L in deionized water, and 2.370 in dechlorinated tap water. In addition to concentration-dependent toxicity, we also observed time-dependent toxicity for ZnO NPs. In addition, the sizes of ZnO NPs increased immediately after dispersion and were 1079 nm, 3209 nm, and 3652 nm in ultra-pure, deionized, and dechlorinated tap water. The highest concentration of measured Zn2+ in exposure concentrations was found in ultra-pure water, followed by deionized and dechlorinated tap water suspensions. Furthermore, Javanese medaka showed high sensitivity to acute exposure of ZnO NPs in all types of water.

Economic contribution and attitude towards alien freshwater ornamental fishes of pet store owners in Klang Valley, Malaysia.

Malaysia is one of the top ten countries in the world that produce freshwater ornamental fishes. This industry can offer better livelihood opportunities to many poor households. However, most of the produced ornamental fishes are alien to Malaysia. In this study, we explore the contribution of alien freshwater fishes to the income of ornamental fish store owners and their attitude towards alien freshwater fishes within Klang Valley, Malaysia. Using a structured questionnaire, we surveyed 70 pet stores out of which 54 (81.42%) store owners responded. Most of the pet store owners were male (72%), Chinese (83%), and the highest educational level was at the secondary level (79%). Most of the pet store owners reported a monthly income of RM 2001-RM 5000 (78%) and were married (73%). Using Chi-square (χ2) test, significant relationships (p < 0.05) existed between the attitude of store owners towards alien ornamental fish species versus educational level (χ2 = 16.424, p = 0.007) and contribution of alien ornamental fishes to the pet store owners' income (χ2 = 27.266, p = 0.003). Fish sales as the main income source also related significantly with the impact of fish selling business on income level (χ2 = 10.448, p = 0.007). This study showed that the ornamental fish sales contributed over half of the income (51-100%) from the businesses of store owners. Almost half of the respondents (42%) reported that alien ornamental fish was the highest contributor to their income from the ornamental fish sale. While the mismanagement of alien ornamental fishes could give various negative ecological impacts, the socio-economic benefits of these fishes cannot be denied.

Reproductive Toxicity of 3,4-dichloroaniline (3,4-DCA) on Javanese Medaka (Oryziasjavanicus, Bleeker 1854).

Compound 3,4-dichloroaniline (3,4-DCA) is a metabolite of several urea herbicides and intermediate chemical of several industrial products. Moreover, 3,4-DCA has been frequently detected in aquatic ecosystems around the world. This aniline is more toxic than the parent chemicals, and it affects non-target organisms. This study evaluated a 21-day reproductive response of an emerging aquatic vertebrate model, Javanese medaka (Oryzias javanicus), exposed to 3,4-DCA. Fecundity and gonads histopathology were observed. The spawning rate and fertilisation reduced significantly in the highest exposed-group (250 µg/L). Gonadosomatic index (GSI) was significantly low in females exposed to 250 µg/L. No substantial structural alteration of male gonads. However, oocyte development and ovarian cell structure were disrupted in 250 µg/L exposed females. The gonadal developmental was not affected in the males; however, a significant reduction in the developmental of female gonads was observed at 250 µg/L. These results show that 3,4-DCA interfere with the reproduction of Javanese medaka through fecundity and alteration of gonadal tissues.

Estimating the Economic Loss Due to Vibriosis in Net-Cage Cultured Asian Seabass (Lates calcarifer): Evidence From the East Coast of Peninsular Malaysia.

This study aims to estimate the economic loss due to vibriosis in the production of Asian seabass in floating net-cages on the east coast of Peninsular Malaysia. Asian seabass has contributed significantly to Malaysia's economic activities and food security. However, its production can be hindered by the occurrence of diseases, such as vibriosis, causing severe economic losses to farmers. A questionnaire-based survey was conducted on 14 small-scale monoculture Asian seabass net-cage farms. Using a stochastic bioeconomic model and inputs from the survey, existing literature, and expert opinion, the economic losses were determined. Moreover, this model considered the prevalence of Vibrio spp. at a farm on the east coast and the risk posed by its infection from hatcheries. The results showed that 71.09% of Asian seabass simulated in the stochastic model survived. The mortality rate due to vibriosis and other causes was at 16.23 and 12.68%, respectively. The risk posed by Vibrio spp. infection from hatcheries contributed to 2.77% of the increase in Asian seabass mortality. The stochastic model estimated that the total cost of producing a tail of Asian seabass was €2.69 per kilogram. The economic loss of vibriosis was estimated at €0.19 per tail per kilogram, which represents 7.06% of the total production cost of Asian seabass per kilogram. An increase in the prevalence of clinical vibriosis and vibriosis case fatality rate at 42 and 100%, respectively, will lead to an increase in the cost of grow-out Asian seabass by €0.29 per tail from the default value. An increase in pellet price per kilogram by €1.38 and feed conversion ratio pellet by 0.96 will consequently increase the cost of grow-out Asian seabass by €2.29 per tail and €0.82 per tail, respectively. We find that the occurrence of Vibrio spp. infection at the hatchery level can contribute to an increased risk in the mortality of Asian seabass during the grow-out phase. Hence, we also need to focus on the control and prevention of vibriosis infection from hatcheries.