Search Results (10)

Managing fishery resources is crucial to ensure the marine environment continues to provide diverse goods and services. To overcome difficulties of classical methods used for fish stock management, molecular tools have shown potential to address this issue assessing both targeted and non-targeted species. This study aims to evaluate the spatiotemporal diversity of fish using 12S rRNA gene eDNA metabarcoding sequencing in the Berlengas archipelago and compare two seawater eDNA sampling sources: samples collected by fishermen during their activities and those collected by our research team. The results indicated that autumn presented the highest diversity and that the area around Berlenga Island was the richest area, increasing biodiversity across the region. Fisher-collected samples were generally less diverse than those by the research team but detected species typical of deeper and open-ocean habitats, validating this sampling method. Our study also highlighted eDNA’s role in monitoring fish species by detecting unexpected species for the region, such as Atlantic salmon (Salmo salar) and Atlantic cod (Gadus morhua), while cautioning against false positives like orange clownfish (Amphiprion percula) and blue tilapia (Oreochromis aureus). Future optimisation of our eDNA sampling methodology could better refine marine ecosystem dynamics around the UNESCO Biosphere Reserve of the Berlengas Archipelago, Portugal. Full article

Aquaculture of ornamental marine fish is often conducted in recirculating aquaculture systems (RAS) using artificial seawater. Considering the cost of salts to produce artificial seawater (salinity 35‰), we investigated the effect of different salinities (5, 15, 25, and 35‰) on survival, growth, and oxidative stress responses of juvenile clownfish Amphiprion ocellaris. All fish died when reared at salinity 5‰, but at all other salinities survival was ≥95% in the other treatment groups. There was no influence of salinity on growth and oxidative stress responses of clownfish reared at salinities 15, 25, and 35‰, except for the activity of glutathione S-transferase (GST) of fish reared at 25‰, which was significantly lower compared to those reared in salinity 35‰. The salinity of home aquariums is usually 35‰, so even though clownfish can be reared in brackish water, they need to be transferred to full strength seawater (35‰) in order to be commercialized. Therefore, we also evaluated the responses of acute transference of fish reared at 15 to salinity 35‰. There were no mortalities associated with acute salinity transference and no oxidative damage was observed either. The total capacity against peroxyl radicals (ACAP) was immediately increased after fish were placed in salinity 35‰, and remained high after 168 h (7 days), helping fish to deal with oxidative threats. In conclusion, it is possible to rear juvenile clownfish at 15‰ without harming growth or inducing oxidative stress, possibly reducing costs of water salinization. They can be transferred from brackish water to salinity 35‰ immediately before going to the retail market, with no mortality or oxidative damage. Full article

Triphenyltin (TPT), a kind of persistent pollutant, is prevalent in the aquatic environment and could pose a threat to coral reef fish. However, little is known about the toxicity of TPT on coral reef fish, especially regarding the representative characteristics of body coloration. Therefore, this study chose the clownfish (Amphiprion ocellaris) in order to investigate the effects of TPT exposure on its carotenoid-based body coloration under the environmentally relevant concentrations (0, 1, 10 and 100 ng/L). After TPT exposure for 60 d, the carotenoid contents were decreased and histological damage in the liver was found, shown as nuclear pyknosis and shift, lipid deposition and fibrotic tissue hyperplasia. Liver transcriptomic analysis showed that TPT exposure interfered with oxidative phosphorylation and fatty acid metabolism pathways, which related to carotenoids uptake and metabolism. Furthermore, TPT exposure led to oxidative damage in the liver, which is responsible for the changes in the antioxidant capacity of enzymes, including GSH, MDA, POD, CAT and T-SOD. TPT exposure also affected the genes (Scarb1, CD36, Stard3 and Stard5) related to carotenoid absorption and transport, as well as the genes (GstP1 and Bco2) related to carotenoid deposition and decomposition. Taken together, our results demonstrate that TPT influenced carotenoid-based coloration in coral reef fish by disrupting carotenoid metabolism, which complements the ecotoxicological effects and toxic mechanisms of TPT and provides data for the body color biology of coral reef fishes. Full article

In previous studies, natural killer lysin (NK-lysin) emerged as a crucial antimicrobial peptide (AMP) discharged by NK cells and CTLs. The sequence of NK-lysin was cloned and discovered in some fishes, but its function remains unclear. In our study, we obtained a copy of NK-lysin from the spleen of the healthy clownfish (Amphiprion ocellaris; AoNK-lysin) through cloning and proceeded to investigate its potential functions and activities. The findings showed that the AoNK-lysin gene’s open reading frame (ORF) had a length of 465 base pairs (bp) and encoded 154 amino acids (aa), which included a saposin B domain and six cysteine residues that are highly conserved, forming three intrachain disulfide bonds to carry out antimicrobial activity. The AoNK-lysin gene was widely present in different tissues, with the skin showing the highest expression, followed by the eye, intestine, and muscle. Additionally, the expression of AoNK-lysin was significantly upregulated in the immune organs (spleen, gill, intestine, and head kidney) of A. ocellaris after being challenged by Singapore group iridovirus (SGIV). Furthermore, a 399 base pair cDNA sequence that encodes the fully developed peptide of AoNK-lysin was successfully inserted into a secretion plasmid called pPIC9K. Subsequently, a significant amount of the recombinant AoNK-lysin protein was efficiently manufactured using the Pichia pastoris expression system. The antibacterial test demonstrated that the AoNK-lysin protein significantly suppressed the growth of various pathogens, particularly Streptococcus agalactiae, Streptococcus iniae, Salmonella typhi, Shigella sonnei, Pseudomonas aeruginosa, and Aeromonas caviae. The minimal inhibitory concentration (MIC) was found to be 7.81 μg/mL. Further analysis of antiviral assays showed all the viral mRNA of SGIV to be significantly reduced after AoNK-lysin protein stimuli in FHM cells. Collectively, these discoveries indicate that AoNK-lysin exhibits features of both direct pathogen-killing abilities and inhibited virus replication. Full article

This paper is part of a series of studies aimed at understanding the potential exploitation of the biomass of the polychaete worm Sabella spallanzanii (Gmelin, 1791), which is obtained as a by-product of an innovative Integrated Multi-Trophic Aquaculture (IMTA) system. IMTA systems are designed according to an ecosystem approach with the aim to reduce marine monoculture impact while further increasing production via exploitation of valuable by-products. S. spallanzanii can remove large amounts of suspended matter by filtering large volumes of water per hour and performs well as an extractive organism under IMTA; however, it currently lacks any economic value, thus hindering its sustainable large-scale implementation. However, S. spallazanii has the potential to become competitive as a newcomer in fish bait, as an ornamental organism, and in fish feed markets. Notably, sabella meal has already been successfully tested as an attractant in an innovative fish feed. Here, we refer to the use of sabella meal as the main component (60%) in the formulation of a novel aquarium fish feed. Following the biochemical analysis of farmed sabella meal, the experimental feed was formulated by adding spirulina (25%) and dry garlic (15%) in such proportion as to be isoproteic and isoenergetic to the commercial control feed. After preliminary observations of the palatability of sabella meal for several tropical fish species, the novel experimental feed was tested on ocellaris clownfish, Amphiprion ocellaris (Cuvier, 1830), by evaluating their growth response in a 70-day feeding trial. The fish seemed to enjoy the experimental feed at least as much as the control, and both the control and treatment groups showed no significant differences in weight gain (p = 0.46), specific growth rate (p = 0.76), and feed conversion ratio (p = 0.48), reinforcing the suitability of S. spallanzanii as a viable source of animal proteins to be employed in the fish feed industry in a circular economy perspective. Full article

Lymphocystic disease affects over 150 species of marine and freshwater fish worldwide. In this study, the lymphocystis pathogen was found in 2 (Amphiprion ocellaris and Amphiprion clarkii) of the 9 species of clownfish. Detection of lymphocystis disease virus (LCDV) was based on histopathological study, electron microscope observation of virus particles and gene sequence analysis from the MCP region. Infected A. ocellaris hosts showed sparse, multifocal, white, stiff, papilloma-like nodules on the body, skin, gills and fins; while, on A. clarkia, nodules were found on the operculum skin. Histopathologic study showed lymphocystic cells with an irregular nucleus, enlarged cytoplasm and intracytoplasmic inclusion bodies surrounded by the cell membrane. The viral particle presents virions 180–230 nm in diameter, hexagonal in shape with an inner dense nucleoid under transmission electron micrographs (TEM). From the ML polygenetic tree, the clownfish LCVD genotype was closely related to the LCDV strain from paradise fish, Macropodus opercularis (KJ408271) (pairwise distance: 92.5%) from China, then followed by the strain from Spain (GU320726 and GU320736) (pairwise distance: 90.8–90.5%), Korea (AB299163, AB212999, AB213004, and AB299164) (pairwise distance: 91.5–80.5%) and lastly Canada (GU939626) (pairwise distance: 83%). This is the first report of lymphocystis disease in A. clarkii in Taiwan. Full article

Sea anemones are predatory marine invertebrates and have diverse venom arsenals. Venom is integral to their biology, and is used in competition, defense, and feeding. Three lineages of sea anemones are known to have independently evolved symbiotic relationships with clownfish, however the evolutionary impact of this relationship on the venom composition of the host is still unknown. Here, we investigate the potential of this symbiotic relationship to shape the venom profiles of the sea anemones that host clownfish. We use transcriptomic data to identify differences and similarities in venom profiles of six sea anemone species, representing the three known clades of clownfish-hosting sea anemones. We recovered 1121 transcripts matching verified toxins across all species, and show that hemolytic and hemorrhagic toxins are consistently the most dominant and diverse toxins across all species examined. These results are consistent with the known biology of sea anemones, provide foundational data on venom diversity of these species, and allow for a review of existing hierarchical structures in venomic studies. Full article

Clownfish can be an excellent research model for investigating the socially-controlled sexual development of sequential hermaphrodite teleosts. However, the molecular cascades underlying the social cues that orchestrate the sexual development process remain poorly understood. Here, we performed a comparative transcriptomic analysis of gonads from females, males, and nonbreeders of Amphiprion clarkii, which constitute a complete social group, allowing us to investigate the molecular regulatory network under social control. Our analysis highlighted that the gonads of nonbreeders and males exhibited high similarities but were far from females, both in global transcriptomic profiles and histological characteristics, and identified numerous candidate genes involved in sexual development, some well-known and some novel. Significant upregulation of cyp19a1a, foxl2, nr5a1a, wnt4a, hsd3b7, and pgr in females provides strong evidence for the importance of steroidogenesis in ovarian development and maintenance, with cyp19a1a playing a central role. Amh and sox8 are two potential key factors that may regulate testicular tissue development in early and late stages, respectively, as they are expressed at higher levels in males than in females, but with slightly different expression timings. Unlike previous descriptions in other fishes, the unique expression pattern of dmrt1 in A. clarkii implied its potential function in both male and female gonads, and we speculated that it might play promoting roles in the early development of both testicular and ovarian tissues. Full article

This study aimed to analyse the integrated system of red seaweed Kappaphycus alvarezii and clownfish Amphiprion ocellaris. Experiments were performed in a 45 L aquarium under the following controlled conditions: temperature 22 °C, dissolved oxygen 5 mg/L, salinity 30, and photoperiod 12 h light. There were 10 fish per aquarium, fed with commercial food. Four treatments were tested (three replicates per treatment): (1) only fish (control); (2) fish with 250 g seaweed; (3) fish with 500 g seaweed; and (4) fish with 750 g seaweed. Weekly seaweed and fish were weighed and measured, and ammonia (NH₃), nitrite (NO₂), nitrate (NO₃⁻), phosphate (PO₄³⁻), and phosphorus (P) were analyzed. The total weight (g) (TW) of fish differed over time (F = 0.00) and between treatments (F = 2.84; p = 0.03). Higher TW (±0.75 g) occurred in treatment 3 and after 14 days did not significantly differ from the end of the experiment (p < 0.01). The total length of fish (cm) did not differ significantly over time (F = 0.64; p = 0.66) or between treatments (F = 0.34; p = 0.79). The relative growth rate of seaweed (RGR) did not differ over time (p > 0.12). There was a significant interaction between the nutrients present in the water by treatment and time (F = 3.59; p = 0.00). The nutrient removal efficiency (NRE) of NH₃ and NO₂ was higher in treatments 3 and 1, respectively, reaching ±30% at 14 days (p < 0.001), and from the 21st day of the experiment, the values were close to zero, with no significant difference between treatments up to 35 days (p > 0.90 for both). The NRE of NO₃⁻ increased by 2% at 28 days of the experiment, with no significant difference between treatments (p > 0.07). The NRE of P was ±25% at 14 days and was not different between treatments 1 and 2. The NRE of PO₄³⁻ reached 14% at 21 days for treatments 1, 2 and 3 (p < 0.001). The control showed lower values for the NRE of NH3, NO₃⁻, P, and PO₄³⁻ compared to the treatments, but the NRE of NO₂ increased in 14 days. It is concluded that a greater abundance of K. alvarezii helps in water quality and contributes to the total weight of clownfish between 14 and 21 days. Full article

French Polynesia is experiencing increasing coral bleaching events in shallow waters triggered by thermal anomalies and marine heatwaves linked to climate change, a trend that is replicated worldwide. As sea surface thermal anomalies are assumed to lessen with depth, mesophotic deep reefs have been hypothesized to act as refuges from anthropogenic and natural disturbances, the ‘deep reef refugia hypothesis’ (DRRH). However, evidence supporting the DRRH is either inconclusive or conflicting. We address this by investigating four assumptions of the DRRH focusing on the symbiotic association between anemones and anemonefish. First, we compare long-term temperature conditions between shallow (8 m) and mesophotic sites (50 m) on the island of Moorea from 2011–2020. Second, we compare the densities of the orange-fin anemonefish, Amphiprion chrysopterus between shallow and mesophotic (down to 60 m) reefs across three archipelagos in French Polynesia. Finally, we compare the percentage of anemone bleaching, as well as anemonefish reproduction, between shallow and mesophotic reefs. We found that the water column was well mixed in the cooler austral winter months with only a 0.19 °C difference in temperature between depths, but in the warmer summer months mixing was reduced resulting in a 0.71–1.03 °C temperature difference. However, during thermal anomalies, despite a time lag in warm surface waters reaching mesophotic reefs, there was ultimately a 1.0 °C increase in water temperature at both 8 and 50 m, pushing temperatures over bleaching thresholds at both depths. As such, anemone bleaching was observed in mesophotic reefs during these thermal anomalies, but was buffered compared to the percentage of bleaching in shallower waters, which was nearly five times greater. Our large-scale sampling across French Polynesia found orange-fin anemonefish, A. chrysopterus, in mesophotic zones in two high islands and one atoll across two archipelagos, extending its bathymetric limit to 60 m; however, orange-fin anemonefish densities were either similar to, or 25–92 times lower than in shallower zones. Three spawning events were observed at 50 m, which occurred at a similar frequency to spawning on shallower reefs at the same date. Our findings of thermal anomalies and bleaching in mesophotic reefs, coupled with mainly lower densities of anemonefish in mesophotic populations, suggest that mesophotic reefs show only a limited ability to provide refugia from anthropogenic and natural disturbances. Full article