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Many species in aquatic environments face increased exposure to oncogenic pollution due to anthropogenic environmental change which can lead to higher cancer prevalence. The mechanistic relationship connecting environmental pollution and cancer is multi-factorial and poorly understood, and the specific mechanisms are so far still uncharacterized. One potential mediator between pollutant exposure and cancer is oxidative damage to DNA. We conducted a study in the field with two flatfish species, European flounder ( Platichthys flesus L.) and common dab ( Limanda limanda L. ) with overlapping distribution and similar ecological niche, to investigate if the link between oncogenic pollutants and cancer described in ecotoxicological literature could be mediated by oxidative DNA damage. This was not the case for flounders as neither polycyclic aromatic hydrocarbon (PAH) bile metabolites nor metallic trace element concentrations were related to oxidative DNA damage measurements. However, dabs with higher PAH concentrations did exhibit increased oxidative damage. High oxidative DNA damage also did not predict neoplasm occurrence, rather, healthy individuals tended to have higher oxidative damage measurements compared to fishes with pre-neoplastic tumours. Our analyses showed that flounders had lower concentrations of PAH bile metabolites, suggesting that compared to dab this species is less exposed or better at eliminating these contaminants.

The stress hormone cortisol, together with antioxidants, melatonin (Mel) and its biologically active metabolites, 5-methoxykynuramines, including AFMK, set up a local stress response system in mammalian skin. Our in vitro study of the European flounder (Platichthys flesus) was designed to examine whether Mel and AFMK would respond to cortisol while a glucocorticoid is added to the incubation medium. The concentrations of cortisol in the incubation medium mimic plasma cortisol levels seen in fish exposed to different types of stresses such as handling, confinement, high density, food-deprivation or air-exposure. We measured Mel and AFMK in skin explants and culture media using high-performance liquid chromatography (HPLC) with fluorescence detection. We also analysed melanosome response (dispersion/aggregation) in the explants subjected to the different treatments. Cortisol stimulated the release of Mel and AFMK from skin explants in a dose-dependent manner. Melanosome dispersion and a darkening of the skin explants were observed after incubation with cortisol. This study is the first to demonstrate the interrelationship between cortisol and Mel/AFMK in fish skin. Our data strongly suggest that the cutaneous stress response system (CSRS) is present in fish. The question remains whether Mel, AFMK or cortisol are synthetized locally in fish skin and/or transported by the bloodstream. The presence of the CSRS should be taken into account during elaboration of new indicators of fish welfare both in aquaculture and in the wild., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Juvenile flounders (Platichthys flesus), collected in two estuaries with similar temperature regimes (the heavily polluted Seine and the moderately contaminated Vilaine), were submitted to a common garden experiment. After an acclimation period, both populations were challenged by a thermal stress (9-24°C for 15days, then maintenance at 24°C for 19days). The condition factor of the Vilaine fish increased in both conditions, while it decreased for the heated Seine flounders after 34days. The expression of genes related to the energetic metabolism was measured in the liver. The expression levels for ATP-F0 and COII were significantly reduced for heated vs. standard fish from both estuaries, while a decrease of the 12S expression was detected only in heated vs. standard fish from the Seine estuary. Thus, it is suggested that highly contaminated fish from Seine could display a lower tolerance to thermal stress, compared to moderately contaminated fish from Vilaine., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

This study represents the first attempt to assess genotoxicity and cytotoxicity effects in herring (Clupea harengus membras), flounder (Platichthys flesus), and cod (Gadus morhua callarias) caught at 47 study stations, located close to chemical munition dumpsites in the Gotland Basin, the Baltic Sea. Herring sampled from stations located in the center of chemical munition dumpsites exhibited the highest levels of micronuclei (MN) and total genotoxicity (ΣGentox), which is defined as the sum of frequencies of such nuclear abnormalities as micronuclei, nuclear buds, nuclear buds on the filament, and bi-nucleated erythrocytes with nucleoplasmic bridges. Exceptionally high and high ΣGentox risks were determined for flounder (89.47%), herring (79.31%), and cod (50%) caught at the stations located close to the chemical munition dumpsites. [ABSTRACT FROM AUTHOR]

Human‐generated pressures are continuing to have a detrimental effect on diadromous fishes, resulting in legislative initiatives to conserve and manage these species. Field studies to inform these initiatives focus almost exclusively on larger marine, estuarine, and inland waters, neglecting the role of small coastal streams and tributaries in population recovery. As an example of the potential contribution of very small coastal streams to diadromous fish recruitment, we report here on the densities and distribution of European eel Anguilla anguilla L. and flounder Platichthys flesus (L.) in one such stream, "La Warenne," in northeast France in October 1998 and 1999. Both species were found mainly in downstream sites in both years—flounder were captured in low numbers only, but mean CPUE values for eel were similar to the highest densities reported in the ICES WGEEL database for larger rivers of Great Britain, France and Spain in the Interreg Atlantic and North Sea areas. This small dataset illustrates that small coastal streams may represent an important, but overlooked, role in conservation strategies for European eel, and possibly flounder, indicating the need for further field studies and investigations of existing data sources to evaluate the contributions to diadromous fish recruitment of these poorly studied water courses. [ABSTRACT FROM AUTHOR]

• The effect of seawater to freshwater transfer (and vice versa) on UI and ion channel expression in the CNSS was examined. • An involvement of UI in adapt transfer from seawater to freshwater in flounder. • Ion channel mRNA expression was lower in CNSS from freshwater-adapted flounder as opposed to seawater. The caudal neurosecretory system (CNSS) is a part of stress response system, a neuroendocrine structure unique to fish. To gain a better understanding of the physiological roles of CNSS in fluid homeostasis, we characterized the tissue distribution of urotensin I (UI) expression in European flounder (Platichthys flesus), analyzed the effect chronic exposure to seawater (SW) or freshwater (FW), transfer from SW to FW, and reverse transfer on mRNA levels of UI, L-type Ca2+ channels and Ca-activated K+ channels transcripts in CNSS. The tissue distribution demonstrated that the CNSS is dominant sites of UI expression, and UI mRNA level in fore brain appeared greater than other non-CNSS tissues. There were no consistent differences in CNSS UI expression or urophysis UI content between SW- and FW-adapted fish in July and September. After transfer from SW to FW, at 8 h CNSS UI expression was significantly increased, but urophysis UI content was no significantly changes. At 24 h transfer from SW to FW, expression of CNSS UI was no apparent change and urophysis UI content was reduced. At 8 h and 24 h after transfer from FW to SW UI expression and urophysis UI content was no significantly effect. The expression of bursting dependent L-type Ca2+ channels and Ca-activated K+ channels in SW-adapted fish significantly decreased compared to those in FW-adapted. However, there were no differences in transfer from SW to FW or from FW to SW at 8 h and 24 h. Thus, these results suggest CNSS UI acts as a modulator in response to osmotic stress and plays important roles in the body fluid homeostasis. [ABSTRACT FROM AUTHOR]

Abstract The study explores the fine scale spatial and temporal distribution of the entire demersal fish and flatfish assemblages in a brackish ecosystem with a special focus on the abiotic and biotic drivers influencing the abundance of the three commercially and ecologically important flatfish species, namely flounder (Platichthys flesus), plaice (Pleuronectes platessa) and dab (Limanda limanda). Using a 12 year time series obtained by a summer survey in the western Baltic Sea, at least three distinct demersal fish species assemblages were identified. Their spatial distribution was relatively stable over time and could be linked to environmental gradients. The demersal fish assemblages were dominated by few species and differed primarily in the percentage of common species. Both, the demersal and the flatfish assemblage were structured according to different habitat preferences. While there were species-specific differences, boosted regression tree modelling (BRT) showed that interannual fluctuations explained a large percentage of the variance in flatfish abundance whereby salinity, water temperature and sediment type were identified as the most important abiotic drivers. Dab was mainly influenced by sediment type and high salinity, while for flounder the main driver was water temperature. Plaice was also impacted by salinity, but was primarily influenced by biotic variables. The availability of benthic prey organisms in the area was verified as biotic driver for flatfish, especially for plaice. The results underline the importance of accounting for fine scale spatial heterogeneity in marine spatial planning in the Baltic Sea and comparable brackish water ecosystems and highlight the importance of considering benthic prey availability and diet analyses in demersal fish monitoring programs. [ABSTRACT FROM AUTHOR]

Frequencies of eight nuclear abnormalities (NAs) reflecting environmental genotoxicity and cytotoxicity, were examined in 739 specimens of herring ( Clupea harengus ), flounder ( Platichthys flesus ) and cod ( Gadus morhua ) collected between 2009 and 2017 at 50 study stations located in the Gulf of Gdansk and the southern part of the Gdansk Deep (the Baltic Sea). The highest levels of geno-cytotoxicity were recorded in fish caught at stations located in close proximity to chemical and conventional munitions dumping sites or in zones polluted by Chemical Warfare Agents (CWAs). The values of NAs were significantly higher (up to 51-fold compared with the reference level) in herring caught at seven stations and in cod from one station located close to the munitions dumping sites. Exceptionally high total genotoxicity (∑Gtox) risk was found for flounder collected from 18 (72.0%) stations, herring caught at 12 (38.7%) stations and cod caught at four (17.4%) studied stations. [ABSTRACT FROM AUTHOR]

Marine teleosts inhabiting the brackish Baltic Sea have adapted to the less saline water with activation of spermatozoa at low salinity hypo‐osmotic conditions but with shorter longevity and lower swimming speed that affect the fertilization capacity. Aiming to elucidate if the fertilization capacity may be maintained by increasing the number of spermatozoa produced, testis size for the euryhaline flounder Platichthys flesus with external fertilization was assessed along a salinity gradient; with spawning at a salinity of c. 7, 10–18 and 30–35. Fulton's condition factor K = 0.881 ± 0.085 (mean ± S.D.), 0.833 ± 0.096 and 0.851 ± 0.086, for fish spawning at salinities of c. 7, 10–18 and 30–35, respectively, with no difference between areas, i.e. analysed fish were in similar nutritional condition. A general linear model, with testes dry mass as the dependent variable and somatic mass as covariate resulted in a significant difference between areas–populations with larger testes for P. flesus spawning at a salinity of c. 7 but no difference between fish spawning at a salinity of 10–18 and 30–35. The result suggests that adaptation by increasing the number of spermatozoa produced may be a key mechanism for marine teleosts spawning in areas with low salinities to sustain the fertilization capacity as shown here for the euryhaline P. flesus. [ABSTRACT FROM AUTHOR]

Biological indices, biomarkers (EROD activity, DNA SB, 1-OH pyrene), and organochlorine contaminants were investigated in flounder collected in the Gulf of Gdańsk during March–December 2011 to describe their seasonal variability and interrelations. Univariate and multivariate statistics were used to evaluate the relations. The EROD activity positively correlated with DNA SB, both negatively correlated with CF, GSI, and HSI, and there was a moderate positive correlation for EROD and DNA SB with 1-OH pyrene. EROD highest activity corresponded to a resting stage of gonad development. DNA SB, highest during spawning, gradually decreased until late autumn. The PCBs, DDTs, HCB, HCHs, dieldrin, and heptachlor levels in muscle tissue were quantified on a wet and lipid basis. In each case, their levels decreased after spawning, fluctuated over the study period indicating that their accumulation was pronouncedly controlled by chemical-specific properties, their levels in prey, and lipid dynamics. [ABSTRACT FROM AUTHOR]

Two populations of flounder ( Platichtys flesus ) with different life history traits inhabit the brackish water Baltic Sea. Both types share feeding areas in coastal waters during summer-autumn but utilise different habitats for spawning in spring, namely offshore spawning with pelagic eggs and coastal spawning with demersal eggs respectively. Fecundity regulation by atresia was assessed as prevalence (portion of fish with atresia) and intensity (calculated as the average intensity of atresia in these fish) during the reproductive cycle following start of gonad development in the autumn up to spawning in spring, and evaluated in relation to fish condition (Fulton's condition factor reflecting energy reserves of the fish) and feeding incidence of the respective population. Peaking in winter (December–February), fecundity regulation was significantly higher for coastal spawning flounder than for flounder spawning offshore. For coastal spawners, the prevalence was 45–90% with an intensity of 6.4–9.3% vs. 0–25% and an intensity of 2.1–3.4% for offshore spawners during winter. Further, fecundity regulation ceased prior to spawning for offshore spawners but continued for coastal spawners. For coastal spawners, the prevalence was 12–29% and an intensity of 2.5–6.1% during spawning. The change in fish condition was strongly related to feeding incidence and differed between populations. As feeding ceased, condition of offshore spawners decreased during winter up to spawning, whereas condition of coastal spawners decreased during autumn but was maintained as feeding started again prior to spawning. Thus, habitat utilisation according to spawning strategy affects the timing of fecundity down-regulation reflecting availability of resources, namely limited food resources in deep areas and higher availability in coastal areas. Offshore spawning flounder display characteristics typical for a capital spawner with ceasing of feeding and oocyte down-regulation well before spawning, whereas coastal spawning flounder can be characterised as intermediate between a capital and income spawner with feeding prior to and during spawning along with continuous fecundity-regulation. [ABSTRACT FROM AUTHOR]

The Baltic Sea is considered as one of the marine areas most exposed to human impacts. A variety of chemical contaminants pose a threat to the habitants. Female flounder ( Platichthys flesus ) collected from three locations in the southern Baltic Sea in February 2010 were examined for biomarkers of exposure to genotoxic agents (DNA damage), AhR-active contaminants (ethoxyresorufin- O -deethylase, EROD activity), and somatic condition indexes. Organochlorine contaminants (OCs) and polycyclic aromatic hydrocarbon (PAH) metabolites were also measured in individual flounder to evaluate the biological responses in the context of contaminant burden. The genotoxicity, mildly exceeding a background level, revealed a significant relationship with mono-ortho substituted PCB ( m-o PCB). Hepatic EROD activity was highly induced, yet showed no association with any of the contaminants measured other than biliary 1-OH pyrene normalized to pigment absorbance. Significant negative relationships were observed for lipid-based OCs and the gonado-somatic index (GSI) as well as for Ʃ m-o PCB concentrations and the condition factor (CF). Principal component analysis (PCA) revealed an overall connection between somatic condition indexes, biomarkers, and chemical variables. Of the three locations, flounder inhabiting the Gulf of Gdańsk had the greatest contaminant burden and appeared to be the most affected. Of great concern is the reduced GSI in this location which can be attributed to the effects of contaminants and warrants further investigation. [ABSTRACT FROM AUTHOR]

Contamination levels and profiles of 7 polychlorinated-p-dioxins, 10 polychlorinated furans (PCDD/Fs) and 12 dioxin-like polychlorinated biphenyls (dl-PCBs) were investigated in juvenile European flounder (Platichthys flesus) captured in different nursery areas in the northeastern Atlantic coast across its geographical distribution range. The toxic equivalent concentrations (WHO-TEQfish) were also determined in order to evaluate which P. flesus population was more exposed to dioxin-like toxicity. Juveniles caught in the Sørfjord (Norway) showed the lowest WHO-TEQfish concentration (0.052pg WHO-TEQfishg−1wet weight) whereas the highest value was observed in fish from the Wadden Sea (The Netherlands; 0.291pg WHO-TEQfishg−1ww), mainly due to the greater contribution of 2,3,7,8-tetrachlorodibenzo-p-dioxin, the most toxic congener. Nonetheless, when comparing the results with existent tissue residue-based toxicity benchmarks, no adverse effects resulting from PCDD/Fs and dl-PCBs are expected to occur in flounder from the studied systems. [ABSTRACT FROM AUTHOR]

Highlights: [•] Flounders were exposed to contaminated sediments in mesocosms for 7months. [•] Little differential accumulation of contaminants was found by chemical analysis. [•] 1-Hydroxypyrene and DNA damage increased in exposed fish. [•] Metabolic and transcriptional changes were detected in fish livers. [•] Molecular alterations were consistent with chronic PAHs exposure. [Copyright &y& Elsevier]

Abstract: The data on environmental genotoxicity and cytotoxicity levels as well as on genotoxicity risk in flounder (Platichthys flesus), herring (Clupea harengus) and cod (Gadus morhua) collected in 2010–2012 at 42 stations located in chemical munitions dumping areas of the southern Baltic Sea are presented. The frequency of micronuclei, nuclear buds and nucleoplasmic bridges in erythrocytes was used as genotoxicity endpoint and the induction of fragmented-apoptotic, bi-nucleated and 8-shaped erythrocytes as cytotoxicity endpoint. The most significantly increased geno-cytotoxicity levels were determined in fish collected near known chemical munitions dumpsites. Extremely high genotoxicity risk for flounder were identified at 21 out of 24 stations, for herring at 29 out of 31 and for cod at 5 out of 10 stations studied. The reference level of genotoxicity was not recorded at any of the stations revealing that in the sampling area fish were affected generally. [Copyright &y& Elsevier]

Highlights: [•] The two estuaries differed markedly in POP levels and profiles. [•] PCA showed a clear distinction between the examined sites and marine regions. [•] The dominant contaminants, PCBs and DDTs, exceeded the PBDE levels by 4–42 fold. [•] POP profiles in fish were not quite closely reflective of those in sediments. [•] None of the POPs exceeded EAC, except for CB-118 in the Vistula River estuary. [ABSTRACT FROM AUTHOR]

Baltic Sea flounder were recently split into two species, the offshore spawner Platichthys flesus and coastal spawner Platichthys solemdali. The two species can only be distinguished based on egg and sperm characteristics and via genetic analyses, which limits the species identification methods of larvae and juveniles to molecular techniques. We investigated whether otolith chemistry could be used as an additional tool to identify flounder to species level. We tested for species-specific differences in otolith multi-elemental signatures and spatial consistency of those differences for the early life stages of flounder in three areas of the central Baltic Sea (ICES SD 24–28), where the distribution of both species overlaps. Otolith chemistry signatures obtained through maternal transfer (i.e. core chemistry) and signatures that reflect the post-hatching phase were not significantly different between species. Species-specific differences at the sub-regional scale were only found for the Latvian coastal survey area for multiple elements (Ba, Cu, Mg, Pb, Sr and Zn), but were insufficiently distinct for reliable species discrimination. Geographic classification of age-0 juveniles to survey area was more successful than classification to species, which was reflected by a spatial trend in otolith Sr:Ca that followed the salinity gradient and higher Mn:Ca and I:Ca for Latvian individuals. Otolith chemistry of early life flounder from the Baltic Sea reflects spatial variability in environmental conditions but does not differentiate between the two flounder species in sympatric habitats. • Otolith chemistry of age-0 Baltic Sea flounder reflects the salinity gradient • The two recently evolved Baltic Sea flounder species cannot be discriminated using age-0 otolith chemistry • Latvian age-0 flounder have more distinct signatures with higher manganese and iodine [ABSTRACT FROM AUTHOR]

Abstract: The ability of 17α-ethinylestradiol (EE2) to elevate vitellogenin levels were investigated in male flounder Platichthys flesus and vitellogenin concentrations in flounders from the Danish coastal environment were determined. Male flounders were exposed to 17α-ethinylestradiol (EE2) via food or water. Average vitellogenin concentrations in the control fish ranged between 25 and 100 ng mL− 1. Exposure to 5.1, 8.1 and 16.8 ng EE2 L− 1 in water and 500 and 5000 ng EE2 kg− 1 body weight (bw) every second day in the food increased the plasma vitellogenin concentration in a concentration and time dependent manner, whereas exposure to 2.7 ng EE2 L− 1 in water for 21 d and 5 and 50 ng EE2 kg− 1 bw for 12 days in the food did not. EE2 could be detected in liver and testes (but not in muscle) after exposure to 8.1 and 16.8 ng EE2 L− 1 in the water and 5000 ng EE2 kg− 1 bw in the food; the highest concentration was 6 ng g− 1 wet weight in liver. The majority of the male flounders collected from nine coastal Danish sites from 1999 to 2004 had vitellogenin concentrations below 100 ng mL− 1, and only at two sites moderate estrogenic inputs were indicated. [Copyright &y& Elsevier]

Highlights: [•] Transcriptional responses of flounder liver to brominated flame retardants were found. [•] These responses were more sensitive to exposure than enzymatic biomarkers. [•] Mitochondrial oxidative phosphorylation and apoptotic transcripts were dose responsive. [•] Vitellogenin and CYP1A mRNA expression were induced at the highest concentration. [•] Exposure to brominated flame retardants adversely affects key biological pathways. [ABSTRACT FROM AUTHOR]

Abstract: In 1988, epizootics of ulcer disease in the flatfish flounder in the Dutch Wadden Sea were reported near freshwater drainage sluices of IJsselmeer Lake, locally affecting up to 38.9% of fish. Other diseases such as fin rot and lymphocystis were less frequent, but followed a similar pattern. Results of follow-up surveys in the Wadden Sea in 1994–2005 confirm previous findings and also show significantly elevated ulcer prevalences at other smaller drainage works. The most likely stress factors that contributed to the development of the epizootics at these sites include osmotic stress, adverse water quality conditions including chemical contaminants, nutritional deficiencies, and obstruction to fish migration. It was shown that discharges of IJsselmeer Lake freshwater in 1988–96 had a wide effect on the prevalence and distribution of ulcers and lymphocystis in the western Wadden Sea. A general reduction in disease prevalence in flounder in the entire Dutch Wadden Sea was observed during 1988–2005, which was most likely due to a general improvement in water quality and locally improved habitat conditions for flounder near drainage sluices. Ulcer prevalences outside the two IJsselmeer Lake sluices (Den Oever and Kornwerderzand) declined in this period from approximately 30% to 10% for medium-sized fish. Other skin diseases have also displayed a downward trend at both sites in recent years, with prevalences falling sharply to below 1%. Elsewhere in the Wadden Sea and the Ems–Dollard estuary, disease prevalences have declined towards natural background levels (<1%). It is concluded that skin diseases, especially ulcers, are useful indicators of environmental quality in the Wadden Sea. [Copyright &y& Elsevier]

Highlights: [•] Female flounder were sampled from the mouths of the two rivers and corresponding open sea sites. [•] Several differences in biomarkers between the sampling sites were found. [•] Salinity and pollution seem to be the main factors that affect the biomarkers. [•] Effects of chronic pollution were found in the mouths of both rivers. [ABSTRACT FROM AUTHOR]

Highlights: [•] Described for the first time the existence of daily rhythms of CNSS hormone expression and secretion in fish. [•] Compared daily profiles of plasma cortisol, osmolality, electrolytes, glucose and lactate in flounder. [•] Revealed the CNSS may co-ordinate daily changes in energy mobilization, nutritional behavior in flounder. [ABSTRACT FROM AUTHOR]

Abstract: Condition indices and metal bioaccumulation of early life stages of juvenile flounder (5–10cm) were determined in three anthropogenic estuaries (the Scheldt, Seine and Loire) and compared to a reference site (the Canche). Significant correlations were found between metal concentrations in sediment and (i) fish liver for Cd, Cu, Ni, Pb, V and Zn and (ii) fish gills for Cd and Mn. Metal accumulation in juvenile flounder from the three anthropogenic estuaries coincided with significantly lower Fulton’s K indices (from 0.99±0.03 to 1.06±0.01mgmm−3) compared to those from the Canche estuary (from 1.02±0.01 to 1.13±0.01mgmm−3). This discrepancy in fish condition index increased with fish size and therefore, strongly depends on the time juvenile spend in estuary. Muscle lipid contents and Triacylglycerol to Sterol ratios were significantly lower in fish collected in the Scheldt (lipid content: 21.3±3.6%), Seine (17.9±19.8%) and Loire (19.5±2.4%) estuaries compared to those originating from the Canche (38.3±4.6%). This study highlights that combined measures of both fish metal contents and condition indices gives a relevant assessment of juvenile fish health growing in anthropogenic estuaries. [Copyright &y& Elsevier]

Physiological and genetic responses of age 0+ Platichthys flesus were investigated in the eutrophicated and moderately contaminated Vilaine estuary, during a 6-month survey. The main objective of this study was to explore the biological responses of fishes during their juvenile period in an estuarine system in order to detect a possible selective pressure induced by the environmental stress. Our results showed a general convergence in physiological responses along the survey: an increase in genotoxicity was associated with an increase in mRNA expression of ATPase and betaine homocysteine methyltransferase. These results could suggest an increase of cellular damage, energetic request, and detoxification rate related to the growing exposure time to stress. Considering the aging of the cohort, the genetic characteristics of the Vilaine flounder cohort came closer to the one observed in a highly stressed system, the Seine estuary, suggesting a potential selective pressure mainly induced by the chemical stress. [ABSTRACT FROM AUTHOR]

Abstract: Flounder (Platichthys flesus) is a temperate marine fish that is well adapted to the brackish waters of the Baltic Sea. There are two sympatric flounder populations in the Baltic Sea, pelagic and demersal spawners, which differ in their spawning habitat and egg characteristics. In the present study, pelagic spawning flounder of the central Baltic Sea was studied. We examined whether variations in hydrological regime can explain fluctuations in flounder early life stages that have occurred over the past 30years (1970–2005). Using generalized additive modeling to explain the abundance of flounder eggs and larvae in a Latvian ichthyoplankton dataset, we evaluate the hypothesis that the available reproductive volume, defined as the water column with dissolved oxygen larger than 1ml/l and salinity between 10.6 and 12PSU, affects the survival of flounder ichthyoplankton and determines recruitment success. Both reproductive volume and spawning stock biomass were significant factors determining flounder ichthyoplankton abundance. Different measures of water temperature did not contribute significantly to the variability of eggs or larvae. However, recruitment did not correlate to the supply of larvae. The findings presented in this study on the relationship between flounder reproduction, spawning stock biomass and reproductive volume, as well as the lack of correlation to recruitment, are valuable for the understanding of flounder ecology in the Baltic Sea, and for developing the management of the species. [Copyright &y& Elsevier]

The spermatologic characteristics of European flounder (Platichthys flesus luscus) were determined. Flounder were collected during the spawning season and sperm of eight males was extracted by abdominal massage. Body weight and total length of the flounder were measured, volume, motility, duration of motility, spermatocrit, density, and pH of the sperm were determined, and correlations between the spermatologic characteristics and weight and length were investigated. Mean values were 0.7±0.16 ml for sperm volume, 87.5±3.66% for motility, 22.0±1.49 min for duration of motility, 94.0±1.22% for spermatocrit, 2.7±0.16 x 109/ml for density, and 6.9±0.05 for pH. Body length and sperm volume had positive correlations with body weight (p<0.01), but the correlation between length and sperm volume was negative (p<0.01). Likewise, the correlations between spermatocrit and total length, sperm volume, and density were negative (p<0.05). [ABSTRACT FROM AUTHOR]

Abstract: Concentrations of seven polybrominated diphenyl ether (PBDE) congeners were examined in flounder (Platichthys flesus) and sediment in three southern Baltic Sea sites, representing a range of exposure conditions, in order to evaluate spatial differences in PBDE contamination. Additionally, PBDEs were measured in muscle, liver, and gonads of flounder from one of the sites in order to examine inter-tissue distribution. Mean muscle Σ7PBDE levels, in the range of 10–21 ng g−1 lipid, showed inter-site differences attributed to the distance from the Gulf of Gdańsk, and were overall lower than reported earlier in herring, sprat, and salmon. Biota sediment accumulation factors (BSAFs) for Σ7PBDE and individual BDE congeners, in the range of 0.5–24.5, were generally consistent with predicted models for persistent hydrophobic halogenated contaminants. Wet weight (wet wt) PBDE levels in muscle and liver, but not in gonads, were related to tissue lipid content and did not correlate with the fish length and weight. These tissues differed in PBDE levels and profiles as a result of varying lipid content and presumably lipid composition and congener-specific physico-chemical properties. [Copyright &y& Elsevier]

Abstract: Environmental genotoxicity was investigated at 82 locations encompassing different regions of the Baltic Sea. Micronuclei (MN) analysis was performed in erythrocytes of 1892 specimens of flounder Platichthys flesus, herring Clupea harengus and eelpout Zoarces viviparus, three of the most common native fish species of the Baltic Sea collected in 2009–2011. MN background levels in fish were determined using data obtained in 2001–2011 from 107 Baltic sites. Extremely high genotoxicity risk zones were found for flounder at 11 stations out of 16 in 2009 and 33 stations of 41 in 2010–2011, for herring, at 5 of 18 stations in 2009 and 20 of 43 stations in 2010–2011, in eelpout only at one out of 29 stations. The sampling stations were restricted mainly to the southern and eastern Baltic Sea offshore zones and in most of them, MN frequencies in flounder and herring significantly exceeded the reference and background levels of micronuclei. This is a first attempt to evaluate the background MN responses, as well as low, high and extremely high genotoxicity risk levels for native fish species. [Copyright &y& Elsevier]

The monthly distribution of flounder, Platichthys flesus luscus, in the east of the Black Sea was examined by analyzing data from research surveys conducted from January to December in 2003. Although the flounder was distributed widely in depths of 10 to 80 m in the east of the Black Sea, the most specimens were found in coastal waters (<40 m deep) and within a temperature range of 8-17°C during survey periods, and a depth preference of juvenile flounder was below 10 m in the area. The average CPUE at a depth of 10-40 m was higher than at other depth layers. The variation in CPUE was not significant monthly, but significant depending on depths. Length frequency distributions of flounder were similar for all seasons, but mean sizes were statistically different in depths. The spawning of the flounder took place under 80 m deep in the area. [ABSTRACT FROM AUTHOR]

Abstract: Diablo (or SMAC) is a protein released from mitochondria following apoptotic stimuli and inhibits the actions of Inhibitors of Apoptosis (IAP) proteins. IAPs regulate the activity of caspases and NFkB, the primary executioners of apoptosis and of inflammation, respectively. Thus, Diablo is important for the regulation of cellular responses to damage. In Northern Europe, statutory governmental marine monitoring programs measure various biomarkers in flounder to indicate biological effects of pollutant exposure. More recently transcriptomic techniques have been applied in flounder to gain a more comprehensive understanding of pollutant effects, and to discover novel biomarkers. In most of these studies utilising flounder, Diablo was amongst the most highly increased transcripts identified. The aim of this study was to further examine piscine Diablo, at the gene level and mRNA level, after exposure to prototypical pollutants, and in flounder caught from polluted environments. The results show that two genes encoding Diablo exist in fish species, and in flounder one of these genes is increased in liver after exposure to polyaromatic hydrocarbons and polychlorinated biphenyls, and also in livers from fish living on contaminated estuarine sediments. Therefore, Diablo measurement has potential as a biomarker of pollutant exposure, and could indicate damaging effects of chemical contaminants. [Copyright &y& Elsevier]

Flounder (Platichthys flesus), collected in late fall of 2009 from four coastal sites in the southern Baltic Sea including the Gulf of Gdańsk (GoG), were investigated for a suite of biomarkers of contaminant effects. The biomarkers included liver histopathologies, which were diagnosed and assessed using commonly applied lesion categories, the size and density of melano-macrophage aggregates (MMAs), expression of proliferating cell nuclear antigen (PCNA) and of cytochrome P450 1A (CYP1A) in liver as well as the size and density of MMAs and density of Perls'-positive cells in the spleen. The prevalence of liver lesions differed among the sites. Most frequently occurring were non-specific and early toxicopathic non-neoplastic lesions. Mean MMA size was in the range of 264–519μm2 and 717–2137μm2 in liver and spleen, respectively, and density was in the range of 6–13 and 15–26MMAmm−2, respectively. Mean density of PCNA-positive hepatocytes was in the range of 300–1281cells mm−2. These histomorphometrical biomarkers correlated positively with the muscle Hg, Σ7PCB, and ΣDDT residues and negatively with the indices of general liver condition. They showed significant differences between the sites, which were in line with the spatial prevalence of liver lesions and CYP1A induction. Overall, the biomarker responses were more pronounced in the GoG sites in comparison to those outside the Gulf, which confirms some earlier results and broadens the knowledge of contaminant effects in the Polish coastal zone of the Baltic Sea. [Copyright &y& Elsevier]

A total of 1926 flounder (Platichthys flesus luscus Pallas, 1811) were caught monthly by bottom trawl in the south-eastern Black Sea from August 1995 to August 1996. Length-weight relationships, age, growth, mortality, condition factor, gonadosomatic index and sex ratio of the samples were investigated. Individuals were composed of 43 (2.23 %) juveniles, 932 (48.39 %) females and 951 (49.38 %) males. Total length and weight of the specimens ranged from 4.5 to 38.2 cm and from 1.61 to 613.96 g. Age distribution of the samples was from 0 to VI. The length-weight relationship was estimated as W=0.012*L2.974 for all individuals, and von Bertalanffy growth parameters were computed as L∞=43.87 cm, W∞=916.87 g, k=0.23 year-1 and t0=-0.59 year. The total (Z), natural (M) and fishing mortality (F) rates, and the exploitation ratio (E) for all individuals were estimated as Z=0.39, M=0.18, F=0.21 and E=0.53, respectively. As a result, it was concluded that there has not been an important fishing pressure on flounder population in the south-eastern Black Sea. The spawning period of the flounder in the south-eastern Black Sea was estimated between October and December. [ABSTRACT FROM AUTHOR]

The article discusses a study of migrations, site fidelity and homing ability of brand-marked flounder Platichthys flesus in the Tamar estuary, South West England from 1976 to 1980. An otter trawl or beam trawls were used to capture flounder and the freeze-branding technique was employed to mark the fish. Majority of female flounder caught at T2 and T9 stations toward the end of the spawning season were females. During the summer, five recaptures of marked flounder were from Plymouth Sound. Results show that the ability of the Tamar fish to home within the estuary and Plymouth Sound is better than their ability to locate their spawning grounds.

Abstract: Two sympatric flounder populations with different reproductive strategies, offshore spawning at 10–20psu producing pelagic eggs and coastal spawning at 5–7psu with demersal eggs respectively, inhabit the brackish water Baltic Sea. Salinity governs the reproductive success by irregular saline water inflows and hence stock abundance and distribution. The potential fecundity (the standing stock of vitellogenic oocytes in the pre-spawning ovary) was assessed for fish sampled at five locations (two for offshore spawners and three for coastal spawners) along the salinity gradient (ICES SD 25, 27/28, 28 and 29). Multiple linear regression analysis resulted in r 2-values of 0.698–0.894 for the respective sampling location with somatic weight or total fish length as the main predictor, and otolith weight (proxy for age) and oocyte density (by gravimetric counting) as additional predictors. Analysis by univariate regressions using GLM revealed significantly higher fecundity for coastal spawning- than offshore spawning flounder (∼69% more oocytes for an intermediate sized fish) but no intra-population differences. Similarly, gonad dry weight was significantly higher for coastal spawning flounder. Further, growth estimations indicated higher growth in offshore spawning- than in coastal spawning flounder suggesting that the coastal spawning population allocate relatively more resources into reproductive growth than somatic, potentially a result of strong selection for high fecundity of flounder producing demersal eggs due to poor egg survival for this spawning strategy. Earlier studies on fecundity of Baltic flounder are reviewed and discussed with consideration of inter-population differences questioning the idea of in general higher fecundity of flounder in the Baltic Sea. [ABSTRACT FROM AUTHOR]

The occurrence of epizoic ciliates on the juvenile flounder, Platichthys flesus L., 1758, collected monthly by fishing net in Sinop's Sarıkum Lagoon Lake during the period from May 2003 to April 2004, was investigated. Five epizoic ciliate species, Trichodina jadranica (Raabe, 1958) Haider, 1964; Trichodina domerguei Wallengren, 1897; Riboscyphidia sp.; Ambiphrya sp.; and Vorticella sp., were found after examination of 296 juvenile flounder. Among these, T. jadranica was the dominant species. The prevalence and mean intensity values of the 5 epizoic ciliates were given as pooled data for 2 groups, namely mobiline ciliates and sessiline ciliates. Mobiline ciliates were commonly found on the gills of juvenile flounders, whereas sessiline ciliates were found only on fins. The highest prevalence and mean intensity levels of mobiline ciliates were recorded in the winter and spring seasons. Sessiline ciliates were absent in winter. The largest-sized juvenile flounder among the established 6 length classes had a higher intensity of mobiline ciliates. This study is the first on the epizoic ciliate fauna present on the juvenile flounder in Turkey. While T. jadranica is a new parasite record, the juvenile flounder is a new host record for T. domerguei in Turkey. [ABSTRACT FROM AUTHOR]

Abstract: In this study, new DNA markers were explored for the flounder Platichthys flesus. cDNA and genomic sequences of the genes encoding the glyceraldehyde-3-phosphate-deshydrogenase (GAPDH), the cytosolic creatine kinase (CK), the prostaglandin D synthase (PGDS) and the betaine homocysteine methyltransferase (BHMT) were characterized. The tumour suppressor p53 gene structure was already described. A PCR–SSCP (Single Strand Conformation Polymorphism) analysis was finally conducted to study the genetic polymorphism of different populations of flounders collected along the French Atlantic coast. Four highly contaminated French estuaries (Seine, Vilaine, Loire and Gironde) were sampled and compared to a reference estuary (Ster) to explore possible selective effect of the environment on specific allelic frequencies. Our results showed that two loci p53 and PGDS, could be potential markers of chemical stress: p53A allele frequency increased in contaminated systems compared to the reference system. In the Vilaine estuary, PGDS polymorphism could be related to pesticide stress. [Copyright &y& Elsevier]

Age reading is a major source of uncertainty in the assessments of fish stocks. The paper presents preliminary results of alternative approaches to estimating the age (year-class) structure of Baltic flounder, Platichthys flesus. The size structure of catches and the weight distribution of otoliths were analysed using Bhattacharya.s method to distinguish between different cohorts (year classes). The traditional counting of annual growth zones on otoliths yielded the greatest number of age groups (12). Age determination by Bhattacharya.s analysis of otolith weight frequencies enabled to distinguish between 7 cohorts. Bhattacharya.s analysis of length frequencies of the same material allowed distinguishing 5 or 6 different groups. Theoretical weight distributions of otoliths, calculated by Bhattacharya.s method for age groups 1 and 6, were not significantly different from the corresponding distributions of weights of otoliths obtained by the method of counting annual rings. The importance of random selection of individuals for the determination of the age structure of catch by annual rings of otoliths is emphasized. [ABSTRACT FROM AUTHOR]

We intended to estimate if shallow inshore waters of the Gulf of Gdańsk can be considered important feeding grounds for the fishes under investigation. We also examined whether competition for food exists between the investigated fishes which co-exist. The study was carried out at Sopot and Chańupy stations from summer 2001 to autumn 2003. The results showed the high importance of Neomysis integer, which exclusively lives in coastal waters, in the diet of all three investigated fish species. It indicates that they utilize this zone as a feeding ground. The obtained dataset also led to the conclusion that fish trophic niches overlap by about 40%. [ABSTRACT FROM AUTHOR]

European flounder responses to the chemical stress were assessed by a comparative approach on four estuaries displaying contrasted patterns of contamination. The contamination typology of the estuaries was investigated by individual measurements of contaminants in fish. Molecular and physiological responses were studied by gene expression, genotoxicity, neurotoxicity and growth rate. Fishes in contaminated estuaries were characterized by high levels of bioaccumulated contaminants, slow energetic metabolism and reduced growth rate, in contrast to the fish responses in the reference site. A seasonal effect was highlighted for contaminated flounder populations, with high PCB levels, high genotoxicity and elevated detoxification rate in summer compared with winter. [ABSTRACT FROM AUTHOR]

The objective of this study was to investigate the pattern of enzymatic activities, environmental genotoxicity and cytotoxicity in flounder, Platichthys flesus, from the Polish coastal area of the Baltic Sea. Fish were sampled in different contaminated sites in the Gulf of Gdansk and in a reference area outside the gulf. The activities of acetylcholinesterase (AChE), glutathione S-transferase (GST), catalase (CAT), alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatine kinase (CK), lactate dehydrogenase (LDH), were studied, as well as the frequency of micronuclei, nuclear buds and fragmentedapoptotic cells. A higher mean activity level of muscular AChE and a lower activity level of hepatic GST were evident in samples taken from the reference site, relative to those found in the gulf. Modeled CAT activity (in both liver and gill tissue), blood plasma LDH and CK activities were all significantly higher in flounder collected at locations within the Gulf of Gdansk than at the reference site. No statistically significant alterations were observed in the activities of ALT and AST in the blood plasma of flounder in this study. Fish collected from a location at the mouth of the Vistula River showed the highest hepatic GST and CAT, the highest gill CAT activity, and the highest frequency of blood micronuclei, nuclear buds and fragmented-apoptotic cell inductions, as well as the lowest level of blood plasma CK. The present study confirms that compared to fish from the reference area, flounder from the Gulf of Gdansk clearly demonstrate a different enzyme activity, genotoxicity and cytotoxicity biomarker response pattern. [ABSTRACT FROM AUTHOR]

Abstract: The brackish, bloom-forming cyanobacterium Nodularia spumigena produces a peptide called nodularin, which may induce liver damage in fish. In the summer of 2007, nodularin was detected in liver tissue of European flounder caught in Swedish waters of Öresund, within the upper salinity limit for N. spumigena. Nodularin concentrations ranging between 22 and 557μgkg−1 liver (d.w.) were detected in fish liver. Nodularin was not detected in blue mussels (Mytilus edulis). Although N. spumigena blooms can occur in the area, the cyanobacteria were only present in very small amounts in 2007. Results suggested that nodularin accumulated in flounder livers during the summer of 2006, when vast N. spumigena blooms were observed in Öresund, and persisted over several months. Nodularin has previously been shown to induce oxidative stress in mice, crustaceans and mollusks but work on the potential negative effects of nodularin on fish is still scarce. To examine the dynamics of nodularin induced oxidative stress in liver tissue of flounder, the differential responses of the antioxidant enzymes glutathione-S-transferase catalase (CAT) and the formation of malondialdehyde (MDA) were monitored during 14 days in flounder exposed to an intraperitoneal injection of nodularin (0, 2, 10 and 50μgnodularinkg−1 body weight). The activities of GST and CAT in the liver decreased significantly in the 50μgnodularinkg−1 exposure after 7 days, but were restored to control levels after an additional 10 days of recovery. The results suggested that nodularin induced oxidative stress in terms of decreased GST and CAT activity, which can result in increased vulnerability of the cell to reactive oxygen species (ROS). No significant changes could be found in MDA levels between the treatments. Thus, the antioxidant defense system presumably managed to prevent oxygen mediated toxicity as seen by the unchanged levels of MDA. Alteration of the enzymatic defense system may increase energetic costs, thus reducing fish growth and survival. The present study also suggests that oxidative stress biomarkers can be used in fish to detect early responses to nodularin. [Copyright &y& Elsevier]

Abstract: Genomic technologies offer opportunities to gain a more global assessment of the health status of an organism through an understanding of the functional pathways that are responding to pollutant exposure. We have developed a 13,000 clone cDNA toxicogenomics microarray for Platichthys flesus, the European flounder (EU-GENIPOL Project). We aimed to distinguish the origins of flounder taken from six sampling sites of different pollution status in Northern Europe according to their hepatic gene expression profile using bioinformatic approaches. To determine which gene expression differences may relate to pollutant impact, we have completed complementary laboratory exposures of flounder to selected toxicants and determined the associated gene expression profiles. Using multivariate variable selection coupled with a statistical modelling procedure (GALGO) we can predict geographical site but the accuracy is limited to specific sites. The search space for a combination of genes that effectively predicts class membership is very large, however, by combining the signatures derived from acute laboratory exposure to individual chemicals to limit the search space, a very accurate model for classification of all the different environmental sites was achieved. The final model utilised the expression profiles of 16 clones and validation with a qPCR array comprising these genes correctly assigned the site of origin for fish obtained from three of the sites in an independent sampling. These data would imply that the gene expression fingerprints obtained with these arrays are primarily attributable to variations in chemical pollutant responses at the different sites, indicating their potential utility in environmental impact assessment. [Copyright &y& Elsevier]

Abstract: The temporal transcriptomic responses in liver of Platichthys flesus to model environmental pollutants were studied over a 16-day time span after intraperitoneal injection with cadmium chloride (50μg/kg in saline), 3-methylcholanthrene (25mg/kg in olive oil), Aroclor 1254 (50mg/kg in olive oil), tert-butyl-hydroperoxide (5mg/kg in saline), Lindane (25mg/kg in olive oil), perfluoro-octanoic acid (100mg/kg in olive oil) and their vehicles, olive oil (1ml/kg) or saline (0.9%). Statistical, gene ontology and supervised analysis clearly demonstrated the progression from acute effects, biological responses to and recovery from the treatments. Key biological processes disturbed by the individual treatments were characterised by gene ontology analyses and individual toxicant-responsive genes and pathways were identified by supervised analyses. Responses to the polyaromatic and chlorinated aromatic compounds showed a degree of commonality but were distinguishable and they were clearly segregated from the responses to the pro-oxidants cadmium and the organic hydroperoxide, as well as from the peroxisomal proliferator, perfluoro-octanoic acid. This study demonstrated the utility of the microarray technique in the identification of toxicant-responsive genes and in discrimination between modes of toxicant action. [Copyright &y& Elsevier]

Abstract: Previous studies have shown roles for cortisol and prolactin in osmoregulatory adaptation to seawater and freshwater, respectively, in euryhaline fish. This study of the European flounder investigated the potential for these hormones to modulate activity of the caudal neurosecretory system (CNSS), which is thought to be involved in physiological adaptation to changing external salinity. Superfusion of isolated CNSS with either cortisol or prolactin (10 μM; 15 min) led to changes in firing activity in neuroendocrine Dahlgren cells, recorded extracellularly. Cortisol evoked a modest increase in overall firing activity, with the response delayed by 4 h after treatment. The response to prolactin was short latency, continued to build up over the subsequent 4-h wash period, and comprised increased firing activity together with recruitment of previously silent Dahlgren cells. Immunoreactivity for glucocorticoid and prolactin receptors was localised to Dahlgren cells. The CNSS expression level for glucocorticoid-2 receptor mRNA, measured by Q-PCR, was significantly lower in fish fully acclimated to freshwater, compared to seawater. No differences were seen between these two states for prolactin receptor mRNA expression. These results provide evidence for a modulatory action of both hormones on the neurosecretory function of the CNSS. [Copyright &y& Elsevier]

Selection of substrata representing habitats with or without filamentous vegetation by juvenile flounder Platichthys flesus was investigated in laboratory experiments and a field study to assess the effect of the increasing occurrence of ephemeral filamentous macroalgae in shallow nursery areas and its potential effect on flounder recruitment. In the laboratory experiments, 79% of juvenile flounder (24–99 mm total length, LT) preferred bare sand substrata in comparison to substrata with simulated filamentous vegetation. Substratum preference did not change with fish size. Field sampling using a drop trap (1 m2) in a vegetated (>50% coverage of ephemeral macroalgae) habitat in June, July and August showed small flounder (mean LT= 37·3 mm) were caught more often in samples with less vegetation. In contrast, larger flounder (mean LT= 57·2 and 79·7 mm in July and August, respectively) were more evenly distributed in relation to the distribution of vegetation. The results of this study indicate that the increase in ephemeral macroalgae observed in recent years potentially reduces the quantity and quality of suitable nursery habitats for juvenile flounder, which could have detrimental consequences to the overall recruitment of flounder in coastal areas. [ABSTRACT FROM AUTHOR]

Abstract: During the late 17th century (1675–1696), which represents part of the coldest period of the Little Ice Age (known also as the Late Maunder Minimum), fishing took place at more than 20 localities along the coast of the Gulf of Riga. Tax records of the Riga Treasury College indicate that herring (Clupea harengus membras), flounder (Platichthys flesus) and eelpout (Zoarces viviparus) were the major target species at this time. The mean annual catch estimated as landing to market of these three major species was 122tonnes (range 44–230), with an average herring contribution of 73%. Fish catches fluctuated substantially both at temporal (seasonal, annual) and spatial (sub-regional) scales. We suggest that the herring fishery was affected by climatic conditions: during the period of very severe winters (i.e., 1685–1696) landings peaked in the warm summer months and were significantly lower compared to the earlier period of generally higher climate variability and less severe winters (1675–1683). Socio-economic drivers were presumably responsible for the dynamics of the flounder and eelpout fisheries. [Copyright &y& Elsevier]

Nakhlé, K., Cossa, D., Claisse, D., Beliaeff, B., and Simon, S. 2007. Cadmium and mercury in Seine Estuary flounders and mussels: the results of two decades of monitoring. – ICES Journal of Marine Science, 64: 929–938. The flounder (Platichthys flesus) is a flatfish that inhabits marine coastal environments, especially estuaries. It is an alternative quantitative biological indicator to the common marine mussel (Mytilus spp.), which is currently used as a sentinel species to monitor chemical contamination in numerous monitoring programmes. Findings from two decades of monitoring cadmium (Cd) and mercury (Hg) using both sentinel species in the Seine Estuary (France) are reported. For comparison, time-series of water concentrations for the same two metals at the mouth of the River Seine are given. Cd concentrations in the liver of the fish and in the soft tissue of mussels show similar temporal trends, consistent with the major temporal variations of Cd concentrations recorded in river water and with changes in industrial discharge of Cd (phosphogypsum waste) within the Seine Estuary. On the other hand, Hg concentrations in the muscles of flounders show temporal variations with no link to that observed in mussels or fluvial Hg contributions, which are in fact nearly covariant. It is concluded that optimization of the use of flounders as sentinel organisms for monitoring temporal trends of metal contamination in estuarine environments requires in-depth knowledge of its ecology within the area studied. An adapted sampling strategy based on this knowledge should provide results that are easier to interpret. [ABSTRACT FROM AUTHOR]