Assessment of sustainable baits for passive fishing gears through automatic fish behavior recognition.

Low-impact fishing gear, such as fish pots, could help reduce human's impact on coastal marine ecosystems in fisheries but catch rates remain low and the harvest of resources used for baiting increases their environmental cost. Using black seabreams (Spondyliosoma cantharus) as target species in the Bay of Biscay, we developed and assessed the efficiency of biodegradable biopolymer-based baits (hereafter bio-baits) made of cockles (Cerastoderma edule) and different biopolymer concentrations. Through a suite of deep and machine learning models, we automatized both the tracking and behavior classification of seabreams based on quantitative metrics describing fish motion. The models were used to predict the interest behavior of seabream towards the bait over 127 h of video. All behavior predictions categorized as interested to the bait were validated, highlighting that bio-baits have a much weaker attractive power than natural bait yet with higher activity after 4 h once natural baits have been consumed. We also show that even with imperfect tracking models, fine behavioral information can be robustly extracted from video footage through classical machine learning methods, dramatically lifting the constraints related to monitoring fish behavior. This work therefore offers new perspectives both for the improvement of bio-baits and automatic fish behavior recognition.

Risk assessment of wild fish as environmental sources of red sea bream iridovirus (RSIV) outbreaks in aquaculture.

Red sea bream iridovirus (RSIV) causes substantial economic damage to aquaculture. In the present study, RSIV in wild fish near aquaculture installations was surveyed to evaluate the risk of wild fish being an infection source for RSIV outbreaks in cultured fish. In total, 1102 wild fish, consisting of 44 species, were captured from 2 aquaculture areas in western Japan using fishing, gill nets, and fishing baskets between 2019 and 2022. Eleven fish from 7 species were confirmed to harbor the RSIV genome using a probe-based real-time PCR assay. The mean viral load of the RSIV-positive wild fish was 101.1 ± 0.4 copies mg-1 DNA, which was significantly lower than that of seemingly healthy red sea bream Pagrus major in a net pen during an RSIV outbreak (103.3 ± 1.5 copies mg-1 DNA) that occurred in 2021. Sequencing analysis of a partial region of the major capsid protein gene demonstrated that the RSIV genome detected in the wild fish was identical to that of the diseased fish in a fish farm located in the same area in which the wild fish were captured. Based on the diagnostic records of RSIV in the sampled area, the RSIV-infected wild fish appeared during or after the RSIV outbreak in cultured fish, suggesting that RSIV detected in wild fish was derived from the RSIV outbreak in cultured fish. Therefore, wild fish populations near aquaculture installations may not be a significant risk factor for RSIV outbreaks in cultured fish.

Trophic assessment of red seabream (Pagrus major) as a possible bioindicator of human activities in the South Sea of Korea using stable C and N isotopes.

To assess the impact of sand mining on resource utilization by the red seabream (Pagrus major) and the trophic structure of fish assemblages two years after mining activities, we compared stable isotope ratios (δ13C and δ15N) and isotopic niches between aggregated mining and control sites in April and August 2022. Our results showed no spatial differences in the δ13C and δ15N values of red seabream between the sand mining and control sites, suggesting that the mining did not affect their dietary resources. Furthermore, the considerable overlap among fish consumers suggested that the fish food web in mining areas has trophic functions similar to those in natural habitats after mining activities. Overall, our study enhances our understanding of ecosystem conservation and the ecological-based management of coastal areas.

Bioaccumulation, Bioindication and Health Risk Assessment of Heavy Metals in Cape Horse Mackerel (Trachurus trachurus) and Slinger Seabream (Chrysoblephus puniceus) in the Durban Basin and Cape Vidal, South Africa.

The bioaccumulation of heavy metals (HMs) in marine fish is a growing global concern due to potential human health risks. The study analyzed HM in the muscle tissue, gills, and gut of adult male and female cape horse mackerel and slinger seabream caught in the polluted Durban Basin and pristine Cape Vidal from April 2018 to February 2019. Results revealed interspecific, spatial, and organ-specific variability in HM levels. In the Durban Basin, slinger seabream had bioaccumulation (in mg/kg) of As (2.3 ± 0.2), Cr (2.6 ± 0.2), Ni (2.0 ± 0.1), and Pb (4.1 ± 0.3) while cape horse mackerel had Ni (1.6 ± 0.2), Pb (4.7 ± 0.6), and Zn (52 ± 3.01) exceeding World Health Organization (WHO) regulatory limits. Metal pollution index (MPI) values were also higher in Durban Basin (> 5.13) than Cape Vidal (< 3.32) for both species' muscles. Liver and gills of slinger seabream and gut of cape horse mackerel exhibited higher HM accumulation patterns proportionate to the environmental concentrations, indicating the bioindicative potential of HM pollution by the two species. Risk assessment indicated that both fish species had target hazard quotient > 1 for Cr, and target cancer risk < 10-4 for Pb, implying significant potential non-carcinogenic and carcinogenic health risks associated with fish consumption from the Durban Basin. The study recommends daily consumption limits of 16 g/day for slinger seabream and 14 g/day for cape horse mackerel to ensure health safety. The findings contribute to the understanding of HM pollution in the Durban Basin and provide important information for decision-makers and policymakers in developing effective strategies to mitigate and manage HM contamination in fish populations.

Po-210 activity concentrations in wild and farmed fish from the Aegean Sea and Sea of Marmara and dose assessment to consumers.

Polonium (210Po) is the major contributor (with approximately 90%) to the radiation dose from radionuclides contained in the human diet, and it is mostly associated with seafood. This study presents 210Po activity concentrations in the tissues of 16 fish species from the Aegean Sea and Sea of Marmara. Among all species investigated, the highest 210Po activity concentration was 4450 ± 33 Bq kg-1 dry weight (dw) in the digestive tract of anchovy (Engraulis encrasicolus), and the lowest 210Po activity concentration was 1.3 ± 0.6 Bq kg-1 (dw) in the muscle tissue of the thornback ray (Raja clavata). Significant differences in 210Po concentrations were consistently found among the tissues of fish (P < 0.05). In general, the prominent accumulation of 210Po was observed in the digestive tract and liver while the muscle tissue generally displayed the lower concentrations. Polonium concentrations in the internal organs, such as muscle and liver, were related to the feeding ecology of fish and thus are a consequence of 210Po transfer in the food chain rather than 210Po uptake from water. The average 210Po concentration in fish filet was 54.1 Bq kg-1 dw and to attain the recommended limit for the annual committed effective dose (1 mSv year-1) would require the consumption of 1024 kg of mixed fish filet in 1 year, which is unlikely to happen. The highest 210Po activity concentration in the edible part of fish (filet) was determined in the anchovy (E. encrasicolus) but to reach the 1 mSv year-1 limit would require still the consumption of 7.1 kg year-1 of anchovy filet. Similar size specimens of wild and farmed fish, Dicentrarchus labrax and Sparus aurata, were analyzed to assess the differences in 210Po concentrations. Polonium concentrations in the wild fish were several-fold higher than in farmed specimens, these ones fed with fish feed with 210Po content lower than natural food in the sea. Therefore, the current trend of increasing the consumption of seafood from aquaculture seems to be reducing the radiation exposure to 210Po in the human diet that is considered beneficial to public health.

Ecological risk assessment of aquatic organisms induced by heavy metals in the estuarine waters of the Pearl River.

With the rapid economic development of China's coastal areas and the growth of industry and population, the problem of heavy metal contamination in estuarine waters is increasing in sensitivity and seriousness. In order to accurately and quantitatively describe the current status of heavy metal contamination and identify sensitive aquatic organisms with high ecological risks, five heavy metals in eight estuaries of the Pearl River were monitored at monthly intervals from January to December in 2020, and the ecological risks of aquatic organisms induced by heavy metals were evaluated using Risk quotients (RQ) and species sensitivity distributions (SSD) methods. The results showed that the concentrations of As, Cu, Pb, Hg and Zn in estuaries of the Pearl River were (0.65-9.25) µg/L, (0.07-11.57) µg/L, (0.05-9.09) µg/L, (< 0.40) µg/L and (0.67-86.12) µg/L, respectively. With the exception of Hg in Jiaomen water, the other heavy metals in each sampling site met or exceed the water quality standard of Grade II. The aquatic ecological risks of As, Pb and Hg were generally low in the waters of the Pearl River estuary, but individual aquatic organisms are subject to elevated ecological risks due to Cu and Zn. The content of Zn has a lethal effect on the crustaceans Temora Stylifera, and the content of Cu has a serious impact on the mollusks Corbicula Fluminea and has a certain impact on the crustaceans Corophium sp. and the fish Sparus aurata. Heavy metal levels and joint ecological risks (msPAF) in the Humen, Jiaomen, Hongqimen, and Hengmen estuaries were slightly higher than in other estuaries, and the Yamen estuary had the lowest contration of heavy metals and ecological risk. Research findings can serve as a basis for formulating water quality standards for heavy metals and for protecting aquatic biodiversity in the Pearl River Estuary.

Bioaccumulation and human health risk assessment of some heavy metals in sediments, Sparus aurata and Salicornia europaea in Güllük Lagoon, the south of Aegean Sea.

In the current study, it was aimed to determine the sediment in Sparus aurata (Linnaeus 1758) tissues and Salicornia europaea L. Cu, Zn Fe, Hg, Cd, Pb and Mn concentration distributions in Güllük Lagoon which is located in the south of Aegean Sea and under the influence of anthropogenic activities by means of metal pollution and health indices and to investigate the effects of these metals on public health. The concentration range of Cu, Zn, Fe, Hg, Cd, Pb and Mn in sediment samples was determined as 12.16-26.00, 1.62-2.03, 7.77-8.36, 7.52-16.15, 0.071-0.40, 7.99-13.74 and 12.11-12.63 mg kg-1, respectively. Cu and Hg concentrations in sediment were found to be higher than sediment quality guidelines standards. In addition, according to the enrichment factor (EF), Hg, Cd and Cu were found to show above moderate enrichment. Cu, Zn, Fe, Hg, Cd and Mn concentrations in S. aurata muscle tissue were 1.31 ± 2.30, 1.01 ± 0.24, 3.43 ± 0.75, 2.79 ± 0.85, 0.01 ± 0.01 and 1.80 ± 1.12 mg kg-1, respectively. S. europaea heavy metals (HMs) concentrations were determined as Cu = 10.97 ± 3.20, Zn = 0.74 ± 0.62, Fe = 5.69 ± 0.22, Hg = 9.62 ± 8.84, Cd = 0.53 ± 0.33, Pb = 0.22 ± 0.26 and Mn = 8.61 ± 0.14 mg kg-1. It was seen that Hg in S. aurata muscle tissue and Hg in S. europaea concentrations exceeded the limit values determined for consumption purposes. Target hazard quotient (THQ) and total target hazard quotient (TTHQ) values were found to be < 1 for S. aurata and S. europaea. When all these results were considered, it was determined that the metals that could pose a potential ecological and health risk were Hg, Cd and Cu in the study area.

Chronic effects of environmental concentrations of antifoulant diuron on two marine fish: Assessment of hormone levels, immunity, and antioxidant defense system.

The presence and toxicity of waterborne diuron in aquatic environments pose a severe threat to non-target organisms. However, the chronic impact of diuron in marine fish has been poorly investigated. In this study, we report the chronic effects (30 and 60 days) of environmentally relevant concentrations of diuron (0.1, 1, and 10 µg L-1) on economically important marine fish, red seabream (Pagrus major), and black rockfish (Sebastes schlegelii) by evaluating several parameters, including hormone levels, immunity, hepatic function, and antioxidant defense. Significant decreases in 17ß-estradiol and 11-ketotestosterone levels and gonadosomatic index were observed on day 60 in fish exposed to 10 µg L-1 diuron. Parameters of immunity, such as alternative complement activity, lysozyme activity, and total immunoglobulin levels, were significantly lowered by 60-day exposure to 10 µg L-1 diuron in both fish. Significant decreases in the hepatic enzyme activities of alanine transaminase and aspartate transaminase were observed with an induction of cortisol on day 60 in fish exposed to 10 µg L-1 diuron. Intracellular malondialdehyde and glutathione levels were significantly increased by 10 µg L-1 diuron at day 60 with an increase in the enzymatic activities of catalase and superoxide dismutase. Overall, black rockfish were more sensitive to diuron than red seabream. These results suggest that consistent exposure to environmentally relevant concentrations of diuron is detrimental to the reproduction, immunity, and health of marine fish.

Microplastic ingestion evidence by economically important farmed fish species from Turkey.

Microplastic pollution and its potential impacts on humans become a global concern. This study is the first study examining the microplastic ingestion in the commercially important species Oncorhynchus mykiss Rainbow trout (Walbaum, 1792), Sparus aurata Gilthead seabream Linnaeus, 1758, and Dicentrarchus labrax European seabass (Linnaeus, 1758) from Turkey. The occurrence frequency of microplastic (MP)in the gastrointestinal tract (GIT) was varied between 50 and 63 %. The highest MP abundance in the GIT was estimated in Rainbow trout (1.2 MPs individual-1) followed by European seabass (0.95 MPs individual-1) and Gilthead seabream (0.8 MPs individual-1). Most of the extracted microplastic particles were black (61 %) and blue (27 %) in color and fiber (80 %) in shape. Major identified polymers were polyethylene (25 %), polyester (20 %), polyamide (10 %). Since fish consumption is an important route for MPs, results light up the danger potential for humans. This study will fill the information gap in Turkey and show the necessity of protection measures in aquaculture industry.

Microplastics in fillets of Mediterranean seafood. A risk assessment study.

Microplastics (MPs) are considered as emergent threat to human health. No complete data still exists on MPs presence in fish tissue and their transmission to humans. The present study aims to detect and quantify the presence of MPs (<3 µm) in several edible seafood (Sardina pilchardus, wild and farmed Sparus aurata, Mullus surmuletus, Solea solea and musselMytilus galloprovincialis) from the south coast of Mediterranean Sea. MPs were detected through an innovative extraction method coupled to the SEM-EDX technology. The Estimated Daily Intakes (EDIs) for adults and children for each species were calculated. The higher median level (IQR) of MPs (9.09E+04) was found inM. surmuletus. Conversely, the lower median (IQR) level was observed in S. pilchardus (7.04E+04). The smallest and biggest median (IQR)diameter of MPs (1.8 and 2.5 µm) were identified in M. galloprovincialisand S. solea, respectively. The highest EDIs (25.50E+03; 48.09E+03) arefor ingestion of farmedS. auratarespectively for adults and children. Instead, the lowest EDIs (2.37E+02; 4.48E+02) are due to M. galloprovincialisingestion for adults and children, respectively. Our data should be carefully considered in view of the direct exposure of humans to plastic particles under 3 µm through seafood consumption to better manage the related risks.

A multifaceted assessment of the effects of polyethylene microplastics on juvenile gilthead seabreams (Sparus aurata).

Plastic pollution has become a major environmental and societal concern in the last decade. From larger debris to microplastics (MP), this pollution is ubiquitous and particularly affects aquatic ecosystems. MP can be directly or inadvertently ingested by organisms, transferred along the trophic chain, and sometimes translocated into tissues. However, the impacts of such MP exposure on organisms' biological functions are yet to be fully understood. Here, we used a multi-diagnostic approach at multiple levels of biological organization (from atoms to organisms) to determine how MP affect the biology of a marine fish, the gilthead seabream, Sparus aurata. We exposed juvenile seabreams for 35 days to spherical 10-20 µm polyethylene primary MP through food (Artemia salina pre-exposed to MP) at a concentration of 5 ± 1 µg of MP per gram of fish per day. MP-exposed fish experienced higher mortality, increased abundance of several brain and liver primary metabolites, hepatic and intestinal histological defects, higher assimilation of an essential element (Zn), and lower assimilation of a non-essential element (Ag). In contrast, growth and muscle C/N isotopic profiles were similar between control and MP-exposed fish, while variable patterns were observed for the intestinal microbiome. This comprehensive analysis of biological responses to MP exposure reveals how MP ingestion can cause negligible to profound effects in a fish species and contributes towards a better understanding of the causal mechanisms of its toxicity.

Fishmeal replacement by periphyton reduces the fish in fish out ratio and alimentation cost in gilthead sea bream Sparus aurata.

Aquaculture threatens natural resources by fishing down the sea to supply fishmeal. Alternative protein sources in aquafeeds can provide a solution, particularly those that are waste from other operations and thereby reduce feed production costs. Toward this goal, we examined the waste biomass of marine periphyton from biofilters of an integrated multi-trophic aquaculture (IMTA) system as a replacement for fishmeal in diets of gilthead seabream (Sparus aurata). Four isoproteic (41%) and isolipidic (16.7%) aquafeeds were formulated with increased content of periphyton and a corresponding decrease in fishmeal from 20 to 15, 10, or 0%. The growth and biochemical content of seabream fingerlings (initial body weight 10 g) were examined over 132 days. Replacing 50% of fishmeal by waste periphyton improved feed conversion ratio (1.2 vs. 1.35 in the control diet) without harming fish growth. The complete replacement of fishmeal with periphyton resulted in 15% slower growth but significantly higher protein content in the fish flesh (59 vs. 52% in the control diet). Halving fishmeal content reduced feed cost by US$ 0.13 kg-1 feed and saved 30% in the cost of conversion of feed to fish biomass (US$ 0.58 kg-1 produced fish vs. $0.83 in the control diet). Finally, the total replacement of fishmeal by waste periphyton in the diet reduced the fish in-fish out ratio to below 1 (0.5-0.9) as compared to 1.36 in the control diet. Replacing fishmeal with on-farm produced periphyton minimizes aquaculture footprint through the removal of excess nutrients in effluents and the use of waste biomass to reduce the 'fish in' content in aquafeeds and fish production costs. The present study demonstrates the great practical potential of this dual use of marine periphyton in enhancing the circular economy concept in sustainable fish production.

Assessment of the effect of long-term exposure to microplastics and depuration period in Sparus aurata Linnaeus, 1758: Liver and blood biomarkers.

The constant increase in plastic pollution has attracted great attention in recent years due to its potential detrimental effects on organisms and ecosystems. While the consequences of ingestion of large plastic litter are mostly understood, the impacts resulting from a long-term exposure and a recovery period of microplastics (MPs) are still limited. The aims were to monitor oxidative stress, detoxification and inflammatory biomarkers in liver, plasma and erythrocytes of Sparus aurata exposed during 90 days to low-density polyethylene (LDPE)-MPs enriched diet (10% by weight) followed by 30 days of depuration. Exposure to LDPE-MPs progressively activates the antioxidant and detoxification system and induces an inflammatory response in liver and plasma, whereas no significant changes were observed in erythrocytes. The plasma activities of catalase, myeloperoxidase (MPO), lysozyme and the levels of malondialdehyde (MDA) as maker of lipid peroxidation significantly increased after exposure to LDPE-MPs for 90 days compared to the control group. The activities of all antioxidant enzymes - catalase, superoxide dismutase, glutathione peroxidase and glutathione reductase-, the detoxification enzyme glutathione s-transferase, MPO, the production of reactive oxygen species and the levels of MDA were also significantly increased in liver after MPs exposure. Additionally, all these biomarkers tended to recover during the depuration period, most of them reaching similar levels to those of the control group. In conclusion, the ingestion of a diet containing LDPE-MPs for 90 days induced a progressive increase in oxidative stress and inflammation biomarkers in liver and plasma of S. aurata but not in erythrocytes, which tended to regain control values when not exposed to MPs for 30 days. The present study contributes to a better understanding of the toxic effects of MPs in S. aurata and highlights the usefulness of plasma that can be obtained in a minimally invasive way to monitor these effects.

Selenium and mercury balance in sea bream obtained from different living environments in Turkey: a risk assessment for the consumer health.

The selenium health benefit value (Se-HBV) is the criterion developed for revealing nutritional values or the risks related to mercury (Hg) toxicity which can emerge in relation to consumption of fish. In this study, the conditions of Se and Hg elements in the muscular tissues of sea bream (Sparus aurata) living in different environments in Mugla province were examined. In addition, a benefit/risk assessment was performed in terms of consumer health with the calculation of Se-HBV. In this context, muscular tissues of sea bream obtained from wild species (WS), offshore marine cages species (OS), and soil ponds species (SS) were dissolved with microwave wet digestion and the concentrations of Se and Hg (mg/kg wet weight) were determined by using inductively coupled plasma mass spectroscopy (ICP-MS). According to the obtained results, the highest mean Se level was determined in SS (0.43 ± 0.02 mg/kg) followed by WS (0.35 ± 0.03 mg/kg) and OS (0.34 ± 0.02 mg/kg). The highest mean Hg concentration was detected in WS (0.09 ± 0.02 mg/kg) followed by OS and SS as 0.05 ± 0.01 mg/kg respectively. As a result of risk assessment conducted with Se-HBV, positive values were determined for each one of three sea bream groups.

Heavy metal concentrations in wild and farmed gilthead sea bream from southern Mediterranean Sea-human health risk assessment.

Arsenic, cadmium, copper, lead, and zinc concentrations in the edible part of wild and farmed Sparus aurata from Algerian coastal were determined. The highest concentrations of cadmium (0.0078 mg kg-1 wet weight), copper (0.98 mg kg-1 w.w), and zinc (5.1 mg kg-1 w.w) were recorded in farmed sea bream, whereas the highest arsenic (5.02 mg kg-1 w.w) and lead (0.006 mg kg-1 w.w) levels were registered in wild one. The statistical analysis (Friedman test, p < 0.05) indicated that the origin of fish (wild, farmed in a cage, and farmed in raceway) has relevance to the distribution of metal. Estimated weekly intake of inorganic arsenic, cadmium, and lead for a 72.5-kg person consuming Sparus aurata from Algeria does not exceed 1% of the WHO/JECFA recommended rate for all metals, certainly due to the limited consumption of seafood products by the local population. The total target hazard index is far less than "one" 1, and the carcinogenic risk for arsenic exceeds the acceptable value of 10-5. Based on the result of this study, the potential risk to human health from the consumption of contaminate farmed sea bream (in the cage) should be considered.

Assessment of the effects of sex and harvesting season on lipid and fatty acid composition of Sparidae species.

In the present study, the lipid amount and fatty acid profile of different Sparidae species, including gilthead seabream, juvenile gilthead seabream, annular seabream, white seabream, common two-banded seabream were evaluated. Fish were seasonally collected from Köycegiz Lagoon (Mugla, South Western Turkey) from June 2018 to June 2019 and after collection, the sex of each specimen was recorded. According to the results of the study, the highest lipid amount was found in female annular seabream individuals as 8.09 ± 0.78% in November and the lowest lipid amount was found in male juvenile gilthead seabream as 0.98 ± 0.12% in March. Palmitic acid and oleic acid were determined as the most abundant SFA and MUFA for all species, respectively. The highest value of DHA, which was the predominant PUFA was assessed as 15.33 ± 0.26% in female white seabream in November whereas the lowest value (3.83 ± 0.36%) was found in gilthead seabream in December. The n-6/n-3 ratio was determined between 0.27 ± 0.00 (for male common two-banded seabream in July)-1.20 ± 0.03 (for male gilthead seabream in December) and it followed within the range of healthy values for all species. As a conclusion, it was found that values of lipid and fatty acid profiles among the examined Sparidae species vary among the season of collection. The results of the study gave the seasonal nutritional values of four economically-important Sparidae species that being rich in healthy polyunsaturated fatty acids such as DHA, have beneficial in human nutrition.

Alterations of stool metabolome, phenome, and microbiome of the marine fish, red sea bream, Pagrus major, following exposure to phenanthrene: A non-invasive approach for exposure assessment.

The present study aimed to assess the impact of phenanthrene (Phe) on fish health by addressing the alteration of fecal characteristics, in lieu of collecting biomarkers that often involves injurious or even fatal sampling of organisms. The marine fish red sea bream, Pagrus major, was exposed to Phe at a concentration of 18 µg/L for 16 days followed by depuration for 13 days. We collected feces from Phe-exposed or control (Phe-free) fish and then analyzed the fecal metabolite profile (metabolome), carbon utilization of microbiota (phenome), and bacterial 16s rRNA gene sequence (microbiome). Along with the increase in physiological stress markers (SOD and EROD) in serum and liver, we noted the possible role of intestine as a Phe reservoir. Furthermore, abnormal fecal appearance (green coloration) and remarkable changes in fecal characteristics were observed. These changes include alterations of cholesterol and putrescine metabolism and the enhanced utilization of putrescine as a carbon source. Phe also altered the microbial community, with an increase in Phe-degrading bacteria such as Pseudomonas. Interestingly, these enteric impairments were ameliorated by depuration. Taken together, our findings suggest that these alterations in feces were associated with adaptive responses to environmentally relevant Phe exposure scenarios, and that stool samples are potential candidates for exposure assessment in fish.

Helping decision making for reliable and cost-effective 2b-RAD sequencing and genotyping analyses in non-model species.

High-throughput sequencing has revolutionized population and conservation genetics. RAD sequencing methods, such as 2b-RAD, can be used on species lacking a reference genome. However, transferring protocols across taxa can potentially lead to poor results. We tested two different IIB enzymes (AlfI and CspCI) on two species with different genome sizes (the loggerhead turtle Caretta caretta and the sharpsnout seabream Diplodus puntazzo) to build a set of guidelines to improve 2b-RAD protocols on non-model organisms while optimising costs. Good results were obtained even with degraded samples, showing the value of 2b-RAD in studies with poor DNA quality. However, library quality was found to be a critical parameter on the number of reads and loci obtained for genotyping. Resampling analyses with different number of reads per individual showed a trade-off between number of loci and number of reads per sample. The resulting accumulation curves can be used as a tool to calculate the number of sequences per individual needed to reach a mean depth ≥20 reads to acquire good genotyping results. Finally, we demonstrated that selective-base ligation does not affect genomic differentiation between individuals, indicating that this technique can be used in species with large genome sizes to adjust the number of loci to the study scope, to reduce sequencing costs and to maintain suitable sequencing depth for a reliable genotyping without compromising the results. Here, we provide a set of guidelines to improve 2b-RAD protocols on non-model organisms with different genome sizes, helping decision-making for a reliable and cost-effective genotyping.

Assessment of Gill Pathological Responses in Yellowfin Sea Bream (Acanthopagrus Latus) Under Aeromonas Hydrophila Exposure.

Bacterial diseases in cultured fish are considered the main problem with aquaculture system in Iran. The gills are multifunctional organs responsible for respiration, osmoregulation, nitrogenous waste excretion, and acid-base balance. Moreover, they are very sensitive to water contamination. Aeromonas hydrophila (A. hydrophila) is an opportunist pathogen responsible for a wide range of diseases in different species of fish. The gill histological alterations were used to assess the effects of A. hydrophila exposure on yellowfin sea bream, Acanthopagrus latus (A. latus). In this regard, 90 A. latus were exposed to the concentrations of A. hydrophila (103 and 106 CFU/ml) for three weeks. The most histopathological alterations in the gill of the exposed fish included hypertrophy and hyperplasia of the epithelial cells, lamellar fusion, club shaping of gill lamellae, lifting of the epithelium and edema of lamellae with large sub-epithelial space, blood congestion, and hypertrophy and hyperplasia of the mucosal cells. The histopathological alterations were observed in the gill of fish exposed to higher levels of A. hydrophila (106 CFU/ml) consisted of aneurysm and hemorrhage with blood congestion. According to the obtained results of this study, A. hydrophila could cause severe histopathological changes in the gill of A. latus and decrease gas change capability in yellowfin sea bream. Furthermore, the findings of the present study suggested that histopathological changes of the gill provide helpful information about the environmental conditions and as particular biomarkers may help the evaluation of fish general health.

Long-term exposure to waterborne nonylphenol alters reproductive physiological parameters in economically important marine fish.

Low concentrations of nonylphenol (NP) in aquatic environment can induce drastic effects on the endocrine system in animals. In this study, we examined the modulatory effects of NP on reproductive and physiological parameters in juveniles of the red seabream and black rockfish following waterborne NP exposure (0, 1, 10, and 50 µg L-1) for 60 days. In red seabream exposed to 50 µg L-1 NP, plasma levels of 17ß-estradiol (E2) and 11-ketotestosterone (11-KT) were significantly lower at 30 and 60 days, while E2 levels were slightly higher in 10 µg L-1-exposed individuals at day 30. Similarly, significantly lower levels of E2 and 11-KT were observed in 10 and 50 µg L-1-exposed black rockfish at 60 days, whereas the E2 level was higher in 1 µg L-1-exposed individuals at day 30. After exposure to NP, plasma and mRNA levels of vitellogenin (VTG) were significantly higher in both species at 30 and 60 days, similar to the inducible effects from synthetic estrogen. Plasma cortisol levels were significantly elevated by relatively higher concentrations of NP (10 and 50 µg L-1) at 30 and 60 days. Finally, 60 days of exposure of 50 µg L-1 NP significantly decreased the gonadosomatic index (GSI) and increased the hepatosomatic index (HSI) in both species. The results obtained from this study provide an evidence of the endocrine disrupting potential of waterborne NP on early stages of economically important marine fish. The NP-triggered endocrine modulation can induce effects on the development of reproductive and metabolic organs in fish species.