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Nodularin (NODLN) is a cyclic pentapeptide hepatotoxin produced by the cyanobacterium Nodularia spumigena, which occurs regularly in the Baltic Sea during the summer season. In this study flounder ( Platichthys flesus L.) was orally exposed to NODLN either as a single dose or as three repeated doses 3 days apart. Liver and bile samples of the fish were taken 4 days after the last dose. Liver glutathione- S-transferase (GST) activity was also measured and the histopathology of the liver was investigated. The liver of the exposed fish was analyzed by liquid chromatography-mass spectrometry for NODLN concentration. The content of NODLN-like compounds in the bile was analyzed by enzyme-linked immunosorbent assay. NODLN exposure caused slightly incoherent liver architecture and degenerative cell changes in both groups. The mean liver GST activity was significantly higher in the repeatedly dosed flounders than in the singly dosed flounders or in the control. In conclusion, the significantly lower NODLN concentration and the increased GST activity in the liver of the repeatedly dosed flounders compared to the singly dosed flounders suggest that NODLN is rapidly detoxificated. The absence of NODLN glutathione conjugates and the low concentrations of NODLN-like compounds in the bile indicate that detoxification products disintegrate or they are rapidly excreted. [ABSTRACT FROM AUTHOR]

Abstract: Several flatfishes spawn in oceanic waters and pelagic larvae are transported inshore to settle in the nursery areas, usually estuaries, where they remain during their juvenile life. Nursery areas appear as extremely important habitats, not only for juveniles but also for the earlier planktonic larval fish. Yet, the majority of nursery studies tend to focus only on one development stage, missing an integrative approach of the entire early life that fishes spent within a nursery ground. Thus, the present study assessed the influence of environmental parameters on the dynamics of the larval and juvenile flatfishes, throughout their nursery life in the Lima Estuary. Between April 2002 and April 2004, fortnightly subsurface ichthyoplankton samples were collected and juveniles were collected from October 2003 until September 2005. Larval assemblages comprised nine flatfish species, while only six were observed among the juvenile assemblages. Solea senegalensis and Platichthys flesus were the most abundant species of both fractions of the Lima Estuary flatfishes. Larval flatfish assemblages varied seasonally, without relevant differences between lower and middle estuary. Platichthys flesus dominated the spring samples and summer and autumn periods were characterized by an increase of overall abundance and diversity of larval flatfishes, mainly S. senegalensis, associated with temperature increase and reduced river flow. On the contrary, during the winter abundance sharply decreased, as a consequence of higher river run-off that might compromised the immigration of incompetent marine larvae. Juvenile flatfishes were more abundant in the middle and upper areas of the estuary, but the species richness was higher near the river mouth. Sediment type, distance from the river mouth, salinity, temperature and dissolved oxygen were identified as the main environmental factors structuring the juvenile flatfish assemblages. Juveniles were spatially discrete, with the most abundant species S. senegalensis and P. flesus associated with the middle and upper estuary, while the remaining species were associated with the lower estuarine areas. The larval fraction exhibited distinct dynamics from the juvenile estuarine flatfish community. Larval flatfishes showed a strong seasonal structure mainly regulated by biological features as the spawning season and also by seasonal variations of water characteristics. On the other hand, juvenile flatfishes were markedly controlled by site specific characteristics such as sediments structure, distance from the river mouth and salinity regime. The present study emphasized the idea that the environmental control varies throughout the ontogenetic development, stressing the importance of integrating all the early life of a species in flatfish nursery studies. [Copyright &y& Elsevier]

Kijewska, A., Burzyński, A., and Wenne, R. 2009. Molecular identification of European flounder (Platichthys flesus) and its hybrids with European plaice (Pleuronectes platessa). – ICES Journal of Marine Science, 66: 902–906. European flounder (Platichthys flesus) and plaice (Pleuronectes platessa) are commercially important marine fish species inhabiting the continental shelf waters of Europe. Morphological similarity between the two makes it difficult to identify their hybrids, so species misclassification can generate errors in defining stocks in terms of their conservation and management. Flounder and plaice populations from the North Sea and the Baltic Sea were studied. Mitochondrial DNA (mtDNA) was used to confirm the morphological species identification. The set of molecular markers, two mitochondrial (cytochrome b and D-loop) and two nuclear (the ribosomal marker ITS and parathyroid hormone-related protein gene), was constructed to identify the two flatfish species and their hybrids. “Pure” flounder (P. flesus) were observed in the Bay of Gdańsk, Baltic Sea, and off the coast of Denmark in the North Sea. The fishing area near Bornholm in the Baltic is rich in P. flesus × P. platessa hybrids. The length difference of the amplified D-loop fragment was used for species identification. The characteristics of heteroplasmy in the control region (D-loop) can be useful as a population marker in the European flounder. Our studies demonstrate the utility of mtDNA polymorphism combined with nuclear molecular markers for correct identification of the morphologically similar and hybridized European flounder and plaice. [ABSTRACT FROM PUBLISHER]

Management measures to facilitate the recovery of fish stocks can lead to shifts in traditional fishing patterns and target species. In the Baltic Sea, drastic reductions in catch quota for cod (Gadus morhua) force mixed demersal trawl fisheries to avoid cod bycatch and focus on flatfish species. This study developed and tested a simple selection concept that aims to avoid cod bycatch in flatfish-directed trawl fisheries by removing a section of the top panel from the extension piece of the trawl (roofless concept). Sea trials testing the performance of a baseline roofless design, and two designs intended to enhance escape reactions of cod were conducted during two sea cruises. Analysis of the resulting catch data revealed that applying the baseline roofless design consistently reduced cod bycatch by ∼75%. Catches of the target species plaice (Pleuronectes platessa) and flounder (Platichthys flesus) were reduced by less than 15%; however, we estimated that catch losses of the two flatfish species could be balanced by increasing fishing effort to ∼8% and ∼12%, respectively. None of the two roofless designs, intended to enhance escape reactions on cod, improved the performance of the baseline design. Under the scenario of fishery choke caused by limited cod quotas, we estimated that the use of the baseline roofless concept could increase fishing possibilities for flatfish by more than 300%. [ABSTRACT FROM AUTHOR]

Abstract: Survival and growth of the earliest life-stages is considered a key factor in determining the abundance of many marine fish species. For flatfishes, the availability of high quality nursery areas is essential for successful recruitment. Regarding the Baltic Sea, there are large gaps in knowledge on factors that influence the distribution of flatfishes during this sensitive stage. To identify the characteristics of important nursery areas in the Baltic for flounder (Platichthys flesus) and turbot (Psetta maxima), a field survey with push net sampling was conducted in the northern Baltic proper during autumn 2006. The sampling stations were stratified to cover several different habitat types defined by substrate and wave exposure. Apart from density of young-of-the-year (YOY) flatfishes, a number of ecological characteristics of the habitat were recorded. Physical habitat variables included substrate type, salinity, depth, turbidity, vegetation and habitat structure. Variables describing biotic processes, such as prey availability and abundance of competitors, were also sampled. The relationships between the spatial distribution of species and these ecological characteristics were fitted to presence/absence data of juvenile flatfish using generalized additive models (GAM). The best habitat descriptors for flounder in order of contribution were: substrate, habitat structure, salinity, wave exposure and occurrence of filamentous algae. Positive effects of increasing wave exposure, salinity and structure were detected while a high cover of filamentous algae had a negative effect. Sand and gravel were preferred over soft and stony substrates. For turbot the best habitat descriptors in order of contribution were: occurrence of filamentous algae, substrate and turbidity. Turbot showed a preference for areas with a low cover of filamentous algae, high turbidity and sandy substrate. Prey availability and abundance of competitors were not included in the models, indicating that the distribution of flatfishes at the scales studied (tens of kilometres) is mainly governed by physical habitat properties. These results constitute the basis for future efforts on mapping of essential flatfish habitats in the Baltic Sea. [Copyright &y& Elsevier]

Salmonid alphavirus (SAV) is the aetiological agent of pancreas disease (PD), a serious viral disease in salmonids. For several decades, SAV was known to infect salmonid species only, until SAV was detected using real-time PCR in several species of wild-caught flatfishes in Scotland in 2010. The presence of SAV in wild flatfishes has been confirmed by further surveys from Ireland and Scotland. The role of flatfishes in SAV-spread and epizootiology has not been elucidated, and no experimental challenges have been conducted to examine virus tissue tropism, virulence and pathology in flatfishes. Wild-caught flatfishes (common dab; Limanda limanda , European plaice; Pleuronectes platessa , European flounder; Platichthys flesus and lemon sole; Microstomus kitt) were either intramuscularly (i.m.) or intraperitoneally (i.p.) challenged with SAV3 or exposed to SAV3 through cohabitation with i.p. injected salmon. SAV-infections were seen in i.m. and i.p. injected dab and i.p. injected salmon but did not result in a transmissible infection in dab although several routes of entry were assessed (oral route not tested). SAV was detected in several tissues of eight common dab (not from cohabitants), with high SAV-levels in pancreas. No viraemia was detected in the SAV-positive common dab and no virus shedding were detected in the tanks. However, pathology in exocrine pancreas and hearts consistent with SAV-replication were seen. This is the first study reporting SAV-induced pathology in a non-salmonid species. The results from the present challenge study supports evidence for common dab being susceptible hosts for SAV. The study also demonstrates that flatfishes are less susceptible to SAV3-infection than salmon. • SAV-infections were seen in i.m. and i.p. injected common dab, but not in cohabitants. • SAV was detected in several tissues of eight common dab, with high levels in pancreas. • No viraemia was detected in the injected SAV-positive common dab, and no virus shedding were detected in these tanks. • The study supports that pleuronectids (common dab) are susceptible hosts for SAV. • First report of SAV-induced pathology in a non-salmonid species [ABSTRACT FROM AUTHOR]

Several fundamental requirements must be met so that NMR-based metabolomics and the related technique of metabonomics can be formally adopted into environmental monitoring and chemical risk assessment. Here we report an intercomparison exercise which has evaluated the effectiveness of ¹H NMR metabolomics to generate comparable data sets from environmentally derived samples. It focuses on laboratory practice that follows sample collection and metabolite extraction, specifically the final stages of sample preparation, NMR data collection (500, 600, and 800 MHz), data processing, and multivariate analysis. Seven laboratories have participated from the U.S.A., Canada, U.K., and Australia, generating a total of ten data sets. Phase 1 comprised the analysis of synthetic metabolite mixtures, while Phase 2 investigated European flounder (Platichthys flesus) liver extracts from clean and contaminated sites. Overall, the comparability of data sets from the participating laboratories was good. Principal components analyses (PCA) of the individual data sets yielded ten highly similar scores plots for the synthetic mixtures, with a comparable result for the liver extracts. Furthermore, the same metabolic biomarkers that discriminated fish from clean and contaminated sites were discovered by all the laboratories. PCA of the combined data sets showed excellent clustering of the multiple analyses. These results demonstrate that NMR-based metabolomics can generate data that are sufficiently comparable between laboratories to support its continued evaluation for regulatory environmental studies. [ABSTRACT FROM AUTHOR]

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: Maintenance of diversity of life in the sea and supporting sustainable use of marine resources require an ecosystem approach. The principles of differentiation of large geographical units on the ecosystem basis that could be applied in research and for the assessment and management of ecosystems and resources in the Baltic Sea have not been satisfactorily formulated up to now. Long-term data series collected by the authors and literature sources were used to subdivide the Baltic Sea into regional ecosystem units. Ten regions (Kattegat, Belts and the Sound, Arkona, Southwest, Eastern and Northwest of the Baltic Proper, Gulf of Riga, Gulf of Finland, Bothnian Sea, Bothnian Bay) were differentiated, some of them including sub-regions. They were grouped into three macroregions (the Transition Area, Baltic Proper and Large Gulfs). Hydrodynamic and geomorphological characters and the areas of local populations of organisms developed as a result of adaptation of species to their habitats constituted the main basis of the division. Circular cyclonic density-dependent currents in the deeps facilitate levelling of the conditions and integrate ecosystems in the regions. They are also responsible for the establishment of the regional depots of nutrients. The nutrients for the biological production are supplied mainly via the zones of intense vertical mixing (high-energy zones) to the euphotic layer. The regions differ in essential environmental conditions. In the process of adaptation to their habitats fishes (herring Clupea harengus membras, sprat Sprattus sprattus balticus, cod Gadus morhua callarias, flounder Platichthys flesus trachurus etc.) and other organisms have developed infraspecific units. The ecosystems and infraspecific units of exploited species should be assessed and managed separately if their vital parameters significantly differ from one another. [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]

Despite the recent discovery of significant genetic structuring in a large number of marine organisms, the evolutionary significance of these often minute genetic differences are still poorly understood. To elucidate the adaptive relevance of low genetic differentiation among marine fish populations, we studied expression differences of osmoregulatory and stress genes in genetically weakly differentiated populations of the European flounder (Platichthys flesus), distributed across a natural salinity gradient. Flounders were maintained in a long-term reciprocal transplantation experiment mimicking natural salinities in the North Sea and the Baltic Sea. Applying real-time quantitative PCR and microarray analysis we studied expression of four candidate genes (hsp70, angiotensinogen, Na/K-ATPase-α and 5-aminolevulinic acid synthase (ALAS)) in gill, kidney and liver tissues. Genes involved in osmoregulative processes (Na/K-ATPases-α and angiotensinogen) showed highly plastic but similar expression in the two populations dependent on environmental salinity. However, we observed a unique sixfold up-regulation of hsp70 in kidney tissue of flounder from the North Sea following long-term acclimation to Baltic salinities. Similarly, significant differences between North Sea and Baltic flounders in expression of ALAS in response to different salinities were found in gill and liver tissue. These findings strongly suggest that gene expression in flounders is shaped by adaptation to local environmental conditions. This identification of adaptive differences in high gene flow marine organisms adds a new dimension to our current understanding of evolutionary processes in the sea and is of paramount importance for identification, protection and sustainable management of marine biodiversity.Heredity (2008) 101, 247–259; doi:10.1038/hdy.2008.54; published online 18 June 2008 [ABSTRACT FROM AUTHOR]

Physiological mechanisms involved in acclimation to variable salinity and oxygen levels and their interaction were studied in European flounder. The fish were acclimated for 2 weeks to freshwater (1‰ salinity), brackish water (11‰) or full strength seawater (35‰) under normoxic conditions (water Po2 = 158 mmHg) and then subjected to 48 h of continued normoxia or hypoxia at a level ( Po2 = 54 mmHg) close to but above the critical Po2. Plasma osmolality, [Na+] and [Cl−] increased with increasing salinity, but the rises were limited, reflecting an effective extracellular osmoregulation. Muscle water content was the same at all three salinities, indicating complete cell volume regulation. Gill Na+/K+-ATPase activity did not change with salinity, but hypoxia caused a 25% decrease in branchial Na+/K+-ATPase activity at all three salinities. Furthermore, hypoxia induced a significant decrease in mRNA levels of the Na+/K+-ATPase α1-subunit, signifying a reduced expression of the transporter gene. The reduced ATPase activity did not influence extracellular ionic concentrations. Blood [Hb] was stable with salinity, and it was not increased by hypoxia. Instead, hypoxia decreased the erythrocytic nucleoside triphosphate content, a common mechanism for increasing blood O2 affinity. It is concluded that moderate hypoxia induced an energy saving decrease in branchial Na+/K+-ATPase activity, which did not compromise extracellular osmoregulation. [ABSTRACT FROM AUTHOR]

Abstract: Residues of polybrominated diphenylethers (PBDEs), extensively applied as flame retardants, are widely spread in the aquatic environment and biota. The present study investigates effects of the environmentally relevant lower brominated diphenylethers in two fish species in vivo under controlled laboratory conditions. Euryhaline flounder (Platichthys flesus) and freshwater zebrafish (Danio rerio) were exposed to a range of concentrations of a commercial pentabromodiphenylether mixture, DE-71. Chemical analysis of exposed fish showed a pattern of PBDE congeners that was very similar to that in wild fish. The resulting range included environmentally relevant, as well as higher levels. Animals were investigated histopathologically with emphasis on endocrine and reproductive organs. In zebrafish, hatching of embryos and larval development were assessed. Biochemical parameters were investigated in flounder as markers for suggested dioxin-like activity (ethoxyresorufin-O-deethylase=EROD), and activation of endogenous estrogen synthesis (gonad aromatase activity). Thyroid hormones were analyzed in plasma in both species. Benchmark analysis using internal PBDE concentrations showed a mild dose-dependent decrease of hepatic EROD and ovarian aromatase activities, and plasma thyroxin levels in flounder, and an increase of plasma thyroid hormone levels in zebrafish. These trends did not result in statistically significant differences from control fish, and major histopathological changes were not observed. Reproduction in zebrafish appeared to be the most sensitive parameter with statistically significantly reduced larval survival and non-significant indications for decreased egg production at internal levels that were more than 55 times the highest environmental recordings. The present results indicate limited risk for endocrine or reproductive effects of current environmental PBDE contamination in fish. [Copyright &y& Elsevier]

The article presents a study on the use of fin rays to investigate many aspects of fish on the West Coast of Ireland. Pleuronectes platessa and flounder were collected from beaches in Galway and Kerry during the autumns of 2005 and 2006 through beach seining and push netting. It was found that there was no significant difference in fin-ray counts for both plaice and flounder. A positive relationship was noted between the anal rays for flounder and the numbers of dorsal.

We found significant population structure and isolation by distance among samples of flounder (Platichthys flesus) in the Baltic, Kattegat and Skagerrak seas using microsatellite genetic markers. This pattern was almost entirely due to a difference between flounder that have demersal spawning in the northern Baltic, as compared to pelagic spawners in the southern Baltic and on the west coast of Sweden. Among demersal spawners we found neither genetic differentiation nor any isolation by distance among sampling sites. We speculate that demersal flounder are descendants of a population that colonized the Baltic previous to pelagic spawners. The demersal flounder may thus have had longer time to adapt to the low salinity in the Baltic, and accordingly display egg characteristics that make it possible to reproduce at the low salinity levels in the northern Baltic. Among pelagic spawners significant isolation by distance was detected. Pelagic spawners have previously been shown to display clinal variation in egg size, which allows them to float also at the moderate salinity levels up to the region north of the island Bornholm. Management units for harvesting should ideally be based on true biological populations, and for the commercially important flounder up to 15 different management stocks in the Baltic have been suggested. We could not find a population genetic foundation for such a high number of management units, and our data suggest three management units: the northern Baltic (demersal populations), southern Baltic with the Öresund straits and the most northwestern sampling sites (Skagerrak, Kattegat and North Sea).Heredity (2008) 101, 27–38; doi:10.1038/hdy.2008.22; published online 7 May 2008 [ABSTRACT FROM AUTHOR]

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]

The GENIPOL cDNA microarray, comprising some 14 000 elements representing 3336 unique expressed sequence tag clusters, was developed for studies on pollutant chemical effects on hepatic gene expression in the sentinel flatfish species Platichthys flesus, the European flounder. This array derived from probes obtained by targeted cloning of xenobiotic metabolising and responsive genes, subtractive suppressive hybridization of pollutant-treated fish and subtracted normalized cDNA libraries of treated fish has been successfully utilized for diagnosing effects of prototypical pollutants, endocrine disruptors and analysis of environmental effects. In the latter context, it has been recommended by the International Council for Exploration of the Sea that the array is tested in European monitoring programmes for assessment of the biological effects of contaminants. Sequence homology is sufficiently great between P. flesus and other pleuronectid flatfish to enable satisfactory use with other species, including American winter flounder and Japanese flounder, plaice, Atlantic halibut and sole. Elucidation of the effects of pollutants requires knowledge of other affectors of gene expression, and in this context the observed effects of a bacterial infection have been studied. The ‘infection’ was experimentally applied by challenge with a vaccine containing Aeromonas salmonicida, the bacterial agent causing furunculosis in flounder and salmonids. These results show that, despite its currently limited gene coverage, application of this array is not confined to toxicology. [ABSTRACT FROM AUTHOR]

This study represents the first large-scale investigation of IPNV in Scottish wild marine fish. Kidney samples were taken from 30 627 fish comprising 37 species and 45 isolations were made from nine different species, illustrating these as reservoirs of IPNV in Scottish waters. The estimated prevalence of IPNV in the Scottish marine environment was low at 0.15% (90% confidence intervals, (CI) of 0.11–0.19%). This was significantly greater in fish caught less than 5.0 km from IPN-positive fish farms in Shetland, at 0.58% (90% CI of 0.45–0.77%). This prevalence persisted and did not significantly decrease over the 16-month period of study. The estimated prevalence of IPNV for each positive species was less than 1% with the statistically non-significant exceptions of flounder, Platichthys flesus (L.), at 12.5% (90% CI of 0.64–47.06%) and saithe, Pollachius virens (L.), at 1.11% (90% CI of 0.49–2.19%). The 45 isolates were titrated and all but two were below the detection limit of the test (<55 PFU g−1). Titres of 3.8 × 102 PFU g−1 and 2.8 × 101 PFU g−1 were calculated from common dab, Limanda limanda (L.), and saithe, respectively. This study provides evidence that clinical outbreaks of IPN in farmed Atlantic salmon may cause a localized small increase in the prevalence of IPNV in wild marine fish. [ABSTRACT FROM AUTHOR]

Freezing of adult flounder Platichthys flesus and turbot Psetta maxima for 24 h resulted in mean decrease of total length ( LT) of 2·1 and 1·3% and a reduced body mass ( M) of 3·1 and 1·3%, respectively. Increasing the freezing time to 25 days increased the shrinkage in turbot but not in flounder. The LT and MT conversion equations for the different species and time spent frozen are given. The differences between species could be explained by the higher volume to surface area relationship in turbot compared with flounder. [ABSTRACT FROM AUTHOR]

Little is known about local adaptations in marine fishes since population genetic surveys in these species have typically not applied genetic markers subject to selection. In this study, we used a candidate gene approach to investigate adaptive population divergence in the European flounder (Platichthys flesus L.) throughout the northeastern Atlantic. We contrasted patterns of genetic variation in a presumably neutral microsatellite baseline to patterns from a heat-shock cognate protein gene, Hsc70. Using two different neutrality tests we found that the microsatellite data set most likely represented a neutral baseline. In contrast, Hsc70 strongly deviated from neutral expectations. Importantly, when estimating standardized levels of population divergence (FST′), we also found a large discrepancy in the patterns of structuring in the two data sets. Thus, samples grouped according to geographical or historical proximity with regards to microsatellites, but according to environmental similarities with regards to Hsc70. The differences between the data sets were particularly pronounced in pairwise comparisons involving populations in the western and central Baltic Sea. For instance, the genetic differentiation between geographically close Baltic Sea and North Sea populations was found to be 0.02 and 0.45 for microsatellites and Hsc70 respectively. Our results strongly suggest adaptive population divergence and indicate local adaptations at the DNA level in a background of high levels of gene flow, typically found in many marine fish species. Furthermore, this study highlights the usefulness of the candidate gene approach for demonstrating local selection in non-model organisms such as most marine fishes.Heredity (2007) 99, 592–600; doi:10.1038/sj.hdy.6801055; published online 12 September 2007 [ABSTRACT FROM AUTHOR]

The histopathology, ultrastructure, and immunohistochemistry of the alimentary canal of flounder Platichthys flesus (L.), naturally infected with the nematode Anisakis simplex s.l. (Rudolphi 1809) from the River Forth (Scotland), were investigated and described. Eight of the 16 flounders were infected with A. simplex s.l. larvae (L3); parasites were encapsulated by serosa on the external surface of the host's digestive tract (intensity of infection 1-8 parasites per host), although nematode larvae were found encysted under the peritoneal visceral serosa of the host spleen and liver and, occasionally, in the liver parenchyma (intensity of infection 3-10 parasites per host). In all sites, different structural elements were recognized within the capsule surrounding larvae. Among the epithelial cells of the intestine of 5 flounders with larvae encysted on external surface of the gut, the presence of several rodlet cells (RCs) was observed. Furthermore, often the occurrence of macrophage aggregates (MAs) was noticed in infected liver and spleen, mainly around the parasite larvae. Eight neuropeptide antisera were tested with immunohistochemistry methods on gut sections of 4 P. flesus infected with extraintestinal nematodes. Sections from the gut of 5 uninfected flounder were used for comparative purposes. In the tunica mucosa of parasitized P. flesus, several endocrine epithelial cells were immunoreactive to anti-CCK-39 (cholecystokinin 39) and -NPY (neuropeptide Y) sera; furthermore, in the myenteric plexus, a high number of neurons were immunoreactive to antibombesin, -galanin, and several to -NPY and -PHI (peptide histidine isoleucine) sera. [ABSTRACT FROM AUTHOR]

Population structure was previously believed to be very limited or absent in classical marine fishes, but recently, evidence of weakly differentiated local populations has been accumulating using noncoding microsatellite markers. However, the evolutionary significance of such minute genetic differences remains unknown. Therefore, in order to elucidate the relationship between genetic markers and adaptive divergence among populations of marine fishes, we combined cDNA microarray and microsatellite analysis in European flounders ( Platichthys flesus). We demonstrate that despite extremely low levels of neutral genetic divergence, a high number of genes were significantly differentially expressed between North Sea and Baltic Sea flounders maintained in a long-term reciprocal transplantation experiment mimicking natural salinities. Several of the differentially regulated genes could be directly linked to fitness traits. These findings demonstrate that flounders, despite little neutral genetic divergence between populations, are differently adapted to local environmental conditions and imply that adaptation in gene expression could be common in other marine organisms with similar low levels of population subdivision. [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]

A survey was undertaken to identify metazoan ectoparasite species on the European flounder, Platichthys flesus (Linnaeus, 1758), in 4 different locations off the north-central Portuguese coast. Parasites of 7 different taxa were found: Caligus diaphanus, caligus sp., and Lepeophtheirus pectoralis (Copepoda: Caligidae); Acanthochondria cornuta (Copepoda: Chondracanthidae); Holobomolochus Confusus (Copepoda: Bomolochidae); Nerocila orbignyi (Isopoda: Cymothoidae); and praniza larvae (Isopoda: Gnathiidae). Lernaeocera branchialis, a common European flounder parasite in the North and Baltic Seas, was not observed among the surveyed fish. Caligus diaphanus, Caligus sp., and Nerocila orbignyi are new host records. The high prevalence and intensity values recorded for L. pectoralis and A. cornuta suggest that both parasite species are common to the European flounder along the north-central Portuguese coast. In contrast, infection levels with respect to the other parasite taxa were, in most cases, comparatively lower, thereby indicating that they only occur occasionally among flounders in the surveyed area. [ABSTRACT FROM AUTHOR]

An extensive cyanobacterial bloom in the southern Gulf of Finland was observed from the end of June to the middle of July 2002 with a subsequent decaying phase of algae during 2.3 weeks after the bloom collapse. Aphanizomenon was the predominating genus in offshore areas, reaching in abundance the absolute maximum of the period 1997.2004. Mainly dead mass of phytoplankton driven to the coastal waters by wind affected significantly the abundance of 1-year-old flounder. Catch per unit effort of commercial size flounder decreased at several national fishing rectangles that were exposed to strongest cyanobacterial blooms according to satellite images. Strong cyanobacterial blooms were followed by a decrease in the abundance indices of the flounder spawning stock in Estonian waters of the Gulf of Finland. [ABSTRACT FROM AUTHOR]

A number of evolutionary mechanisms have been suggested for generating low but significant genetic structuring among marine fish populations. We used nine microsatellite loci and recently developed methods in landscape genetics and coalescence-based estimation of historical gene flow and effective population sizes to assess temporal and spatial dynamics of the population structure in European flounder ( Platichthys flesus L.). We collected 1062 flounders from 13 localities in the northeast Atlantic and Baltic Seas and found temporally stable and highly significant genetic differentiation among samples covering a large part of the species’ range (global FST = 0.024, P<0.0001). In addition to historical processes, a number of contemporary acting evolutionary mechanisms were associated with genetic structuring. Physical forces, such as oceanographic and bathymetric barriers, were most likely related with the extreme isolation of the island population at the Faroe Islands. A sharp genetic break was associated with a change in life history from pelagic to benthic spawners in the Baltic Sea. Partial Mantel tests showed that geographical distance per se was not related with genetic structuring among Atlantic and western Baltic Sea samples. Alternative factors, such as dispersal potential and/or environmental gradients, could be important for generating genetic divergence in this region. The results show that the magnitude and scale of structuring generated by a specific mechanism depend critically on its interplay with other evolutionary mechanisms, highlighting the importance of investigating species with wide geographical and ecological distributions to increase our understanding of evolution in the marine environment. [ABSTRACT FROM AUTHOR]

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]

Abstract: The extent to which biological systems interact in fish from multi-contaminant areas needs to be understood for full interpretation of monitoring data. This study investigates the interaction between two biomarkers, ethoxyresorufin-O-deethylase (EROD) activity and plasma vitellogenin (VTG) in the European flounder (Platichthys flesus). Flounder were exposed to several waterborne EROD inducers and estrogenic chemicals on their own and in binary combinations. Each experimental exposure was for 10 days. The estrogenic chemicals suppressed PAH-mediated EROD induction. Ethynylestradiol (EE2) and nonylphenol (NP) had threshold concentrations of EROD inhibition similar to those at which they induced VTG production. Estradiol (E2), however, showed an ability to suppress EROD at a concentration much lower than that at which VTG was induced. This established that, although EE2 is a more potent VTG inducer than E2, it is less potent in its ability to inhibit EROD activity. The PAH, dibenz[a,h]anthracene (DbA), showed no effect on the VTG induction caused by EE2 and E2. A small effect was noted with NP at threshold concentrations for VTG induction. Archived data on flounder hepatic EROD activity collected during estuarine monitoring were reassessed in light of the project findings. It is hypothesised that published EROD monitoring data may be an underestimation of effects if it is assumed that estrogen-mediated MFO suppression is occurring in wild populations. A greater understanding of system interaction and other factors, including genetics, that influence biomarker response to contaminants would be required to interpret biomarker monitoring data. [Copyright &y& Elsevier]

The article presents environmental sciences and pollution research into the spatial distribution and seasonal changes in the concentration of polycyclic aromatic hydrocarbon (PAH) metabolites within the bile fluids of Platichthys flesus flounder and Limanda limanda dab caught in the North Sea and Baltic Sea in the period 1997-2004. Two metabolites, 1-hydroxypyrene and 1-hydroxyphenanthrene were determined by means of high performance liquid chromatography (HPLC). PAHs are part of international environmental programs because they lead to increased levels of tumors in fish liver.

Lernaeocera branchialis is a penellid copepod, the adults of which are commonly found on gadoids in the North Atlantic and North Sea. L. branchialis has a severe negative impact on wild gadoids and is a potentially serious pathogen of farmed cod, Gadus morhua. This paper describes an experimental infection by cohabiting the intermediate host, the flounder, Platichthys flesus, heavily infected with juvenile stages of L. branchialis, with hatchery-reared cod in order to study the pathology of the early stages of infection of the final host. Early stages of L. branchialis induce extensive local gill hyperplasia, large intravascular thrombus formation and a moderate cellular immune response in the cardiac and branchial tissues of G. morhua. The parasite appears to migrate within the lumen of the afferent branchial artery, ventral aorta and in turn, that of the bulbus arteriosus. These findings are consistent with the presence of a haematophagous, voluminous and rapidly invasive parasite. [ABSTRACT FROM AUTHOR]

Nodularin (NODLN) is a cyanobacterial hepatotoxin that may cause toxic effects at very low exposure levels. The NODLN-producing cyanobacterium Nodularia spumigena forms massive blooms in the northern Baltic Sea, especially during the summer. We analyzed liver and muscle (edible meat) samples from common eider (Somateria mollissima), roach (Rutilus rutilus L.), and flounder (Platichthys flesus L.) for NODLN-R by liquid chromatography/mass spectrometry (LC-MS) and enzyme-linked immunosorbent assay (ELISA). Thirty eiders, 11 roach, and 15 flounders were caught from the western Gulf of Finland between September 2002 and October 2004. Eiders from April to June 2003 were found dead. The majority of samples were analyzed by LC-MS and ELISA from the same sample extracts (water:methanol:n-butanol, 75:20:5, v:v:v). Nodularin was detected in 27 eiders, nine roach, and eight flounders. Eider liver samples contained NODLN up to approximately 200 μg/kg dry weight and muscle samples at approximately 20 μg/kg dry weight, roach liver samples 20 to 900 μg NODLN/kg dry weight and muscle samples 2 to 200 μg NODLN/kg dry weight, and flounder liver samples approximately 5 to 1,100 μg NODLN/kg dry weight and muscle samples up to 100 μg NODLN/kg dry weight. The NODLN concentrations found in individual muscle samples of flounders, eiders, and roach (1-200 μg NODLN/kg dry wt) indicate that screening and risk assessment of NODLN in Baltic Sea edible fish and wildlife are required for the protection of consumer's health. [ABSTRACT FROM AUTHOR]

Effects of pesticide exposure on the European flounder, Platichthys flesus, were investigated using a suppression subtractive hybridization method (SSH) to identify up- and down-regulated genes after a 30-day exposure to herbicides (a cocktail of atrazine, diuron, and isoproturon, and a single herbicide, glyphosate). A total of 256 expressed gene sequences were identified as having the potential for being differentially expressed, of which 116 could be identified by homology with databased sequences. The metabolic functions with which they are associated include energy production, general metabolism, signaling, transport, immune system, and structure. Expression of 14 of these genes was analyzed in liver, muscle, and gills by reverse transcriptase-polymerase chain reaction (RT-PCR) under experimental conditions (0, 15, and 30 days of exposure) and under field conditions (sampling in two estuaries displaying different levels of pesticide contamination). This study provides a first basis for studying the response of fish to pesticide exposure and allows the characterization of new potential genetic markers of pesticide contamination in the field. [ABSTRACT FROM AUTHOR]

The effects of cyclic and constant hydrostatic pressure on the norepinephrine and epinephrine content in discrete regions of the brain of European flounder Platichthys flesus were studied. After a 14 day exposure to cyclic hydrostatic pressure with a tidal period of 12·4 h and with a maximum peak of 800 kPa (range 200–800 kPa of absolute hydrostatic pressure), fish showed a highly significant decrease in norepinephrine content when compared to control animals held at constant atmospheric pressure. No changes were detected in fish submitted to a constant hydrostatic pressure of 800 kPa. [ABSTRACT FROM AUTHOR]

Abstract: The accumulation of 4-tert-octylphenol and the associated estrogenic effects were studied after a single pulse exposure to flounder Platichthys flesus. 4-tert-octylphenol was administered orally in a single dose of 50mgkg−1 and tissue (liver, muscle and testis) and plasma concentrations of 4-tert-octylphenol as well as plasma vitellogenin were measured 3, 6, 12, 18, 24, 48, 72, 144 and 216h after administration of the dose. Concentrations of 4-tert-octylphenol in plasma and tissues were determined by Liquid Chromatography Mass Spectrometry (LC–MS). 4-tert-octylphenol was detectable in liver, testis, muscle and plasma 3h post administration and an accumulation was observed in liver, muscle and plasma up to 12h and in testis 18h post administration, respectively. The maximum concentrations of 4-tert-octylphenol in liver, muscle and testis were 67, 3.2 and 6.8μgg−1, respectively. An increase in plasma vitellogenin levels was seen 48h post administration and the vitellogenin level continued to increase (from <100ngml−1 to 1.4mgml−1) until the end of the experiment 9 days after the administration of 4-tert-octylphenol. [Copyright &y& Elsevier]

One prerequisite for a successful stocking programme is the choice of an appropriate release site, which would ensure good growth, survival and recruitment to the local fishery. The influence of different salinity regimes on habitat quality for juvenile flounder,Platichthys flesus(L.), was examined in Danish inshore waters using enclosures to study growth and survival. Three locations were chosen: Virksund (V)– constant low salinity at 10‰; Harre Vig (HV)– constant high salinity at 24‰; and Hjerk Nor– variable salinity, 0–25‰. Fish movement was examined, using a diffusion model, at the first two sites. At HV the fish had unrestricted dispersal opportunity resulting in average individual movement of 45 m day−1. Fish at V were restricted in dispersal in one direction because of a dam, about which they would concentrate under certain hydrographical conditions. Growth was highest at the locations with stable salinity and mortality highest at the location with variable salinity. [ABSTRACT FROM AUTHOR]

Experiments performed on isolated intestinal segments from the marine teleost fish, the European flounder (Platichthys flesus), revealed that the intestinal epithelium is capable of secondary active HCO3- secretion in the order of 0.2–0.3 μmol·cm-2h-1 against an apparent electrochemical gradient. The HCO3- secretion occurs via anion exchange, is dependent on mucosal Cl-1, results in very high mucosal HCO3-concentrations, and contributes significantly to Cl- and fluid absorption. This present study was conducted under in vivo-like conditions, with mucosal saline resembling intestinal fluids in vivo. These conditions result in a transepithelial potential of -16.2 mV (serosal side negative), which is very different from the -2.2 mV observed under symmetrical conditions. Under these conditions, we found a significant part of the HCO3 secretion is fueled by endogenous epithelial CO2 hydration mediated by carbonic anhydrase because acetazolamide (10-4 M) was found to inhibit HCO3- secretion and removal of serosal CO2 was found not to influence HCO3- secretion. Reversal of the epithelial electrochemical gradient for Cl-(removal of serosal Cl-) and elevation of serosal HCO3- resulted in enhanced HCO3- secretion and enhanced Cl- and fluid absorption. Cl- absorption via an anion exchange system appears to partly drive fluid absorption across the intestine in the absence of net Na+ absorption. [ABSTRACT FROM AUTHOR]

The length and weight distribution of flounder ( Platichthys flesus luscus Pallas, 1811) were 14.2-34.1 cm and 28.879- 611.0 g, respectively. The samples were distributed between the 0 and VII age groups. Von Bertalanffy growth parameters for flounder were Lt = 41.241[1-e-0.161(t+3.240)] for females, Lt = 33.567[1-e-0.181(t+3.525)] for males and Lt = 40.577[1-e-0.142(t+3.105)] for the sexes combined. Reproduction took place from December to March but peaked in December. Two recruitment periods were determined for flounder: the first was January to July, and the second August to November. The proportions of recruitment in the first and second periods were 68.13% and 31.87%, respectively. [ABSTRACT FROM AUTHOR]

A study was carried out to investigate the diet and feeding strategies of age 0 year juvenile flounderPlatichthys flesusin two different micro-tidal habitats in the nutrient enriched Mariager Fjord on the Danish east coast. Juvenile flounder and benthic macrofauna were sampled monthly from June to October 1999 in a bare sand habitat and a habitat covered by filamentous and mat forming macroalgae. The presence of the‘opportunistic’ macroalgae created a shift in the dominance of surface dwelling prey such as epifaunal amphipods to more infaunal groups such as oligochaetes and polychaetes. The diet of the flounder varied considerably between the two habitats mainly reflecting prey availability relative to their abundance, prey spatial distribution, habitat structure and ontogenetic prey shifts as a function of total length. Flounder in the vegetated site fed on a diverse diet of copepods, polychaetes and oligochaetes, whereas those caught in the bare sand site fed primarily on the amphipodCorophium volutatorwhich was numerically dominant at this site. During the growth season, two diet shifts were observed: from copepods early in the season to macrofauna organisms and, later in the season, the inclusion of more hyperbenthic prey such asMysideaspp;Idoteaspp. and the common gobyPomatoschistus microps. [ABSTRACT FROM AUTHOR]

Instantaneous growth rates of 0-group European flounder,Platichthys flesus, obtained from two 9-day enclosure experiments were used to assess the relative quality of three eelgrass and three bare sand habitats in the Bay of Aarhus, Denmark. Measurements of temperature, dissolved oxygen, salinity, sediment structure and biomass of benthic macrofauna were made during the experiments. Gut content analysis was performed after retrieval of the fish. Growth rates of flounder at two bare sand sites were higher or equal to growth rates at the three eelgrass sites, and at a muddy bare sand site in both experiments. The variation in gut fullness of flounder between vegetated and bare sand sites followed the pattern observed in growth rates, but diverged from the spatial variation in prey abundance, which indicates that there were differences in the feeding efficiency between sites. Overall, habitat complexity, i.e. the presence or absence of vegetation seems to be the most important factor determining the growth rates and, by corollary, habitat quality for juvenile European flounder in the Bay of Aarhus. [ABSTRACT FROM AUTHOR]

This study examined the electrical firing activity of neuroendocrine Dahlgren cells in the caudal neurosecretory system (CNSS) of the euryhaline flounder in vivo. Intracellular recordings revealed generally similar activity patterns and membrane properties to those previously reported in vitro. To investigate the potential role of the CNSS in osmoregulatory adaptation, extracellular, multiunit, recordings compared the activity patterns of Dahlgren cells in fully seawater- and freshwater-adapted fish. The proportion of cells showing bursting (as opposed to phasic or tonic) activity was greater in seawater- than in freshwater-adapted fish, as was the Correlation Index, a measure of the degree of correlation between firing activities of cells recorded simultaneously from the same preparation. Acute transfer of fish from seawater to freshwater gill perfusion led to recruitment of previously silent Dahigren cells and a reduction in Correlation Index; freshwater to seawater transfer increased the Correlation Index. Severing the spinal cord anterior to the CNSS led to an increase in overall Dahlgren cell activity. Electrical stimulation of branchial nerve branches providing input to the brainstem, or tactile (pinch) stimulation of lips or fins, led to a reduction in CNSS activity lasting up to 500 s, indicating the presence of descending modulatory pathways from the brain. These results are consistent with a role for CNSS neuropeptides, urotensins, in supporting survival in a hypertonic, seawater, environment. [ABSTRACT FROM AUTHOR]

Intersex (ovotestis) is the condition whereby an individual possesses oocytes, at varying degrees of severity, within the normal testicular tissue and is regarded as an endpoint of endocrine disruption in male fish. The occurrence of intersex has been linked with exposure to endocrine-disrupting compounds present in the environment, which are thought to mimic the actions of the female sex hormone 17-b-oestradiol. Using stained histological sections of archived cases of intersex from European flounder (Platichthys flesus), this study developed a grading method to assess the extent of the condition. This was termed the ovotestis severity index (OSI). The OSI utilizes the distribution pattern (focal, diffuse, cluster, zonal) and type of oocyte (previtellogenic, cortical alveolar, vitellogenic) to assign a severity rating to each intersex fish. A high OSI refers to fish with testicular material almost completely replaced with mature (vitellogenic) oocytes, and lower scores indicate those fish with small numbers of immature (previtellogenic) oocytes. The development of this index provides pathologists with a robust tool for the grading of the intersex condition in European flounder and potentially in other fish species. [ABSTRACT FROM AUTHOR]

The neuroendocrine Type 1 Dahlgren cells of the caudal neurosecretory system of the flounder display characteristic bursting activity, which may increase secretion efficiency. The firing activity pattern in these cells was voltage-dependent; when progressively depolarized, cells moved from silent (approximately−70 mV), through bursting and phasic to tonic firing (< −65 mV). Brief (10 s) evoked bursts of spikes were followed by a slow after-depolarization (ADP; amplitude up to 10 mV, duration 10–200 s), which was also voltage-dependent and could trigger a prolonged burst. The ADP was significantly reduced in the absence of external Ca2+ ions or the presence of the L-type Ca2+ channel blocker, nifedipine. BayK 8644 (which increases L-type channel open times) significantly increased ADP duration, whereas the Ca2+-activated nonselective cation channel blocker, flufenamic acid, had no effect. Pharmacological blockade of Ca2+-activated K+ channels, using apamin and charybdotoxin, increased the duration of both ADP and evoked bursts. However, action potential waveform was unaffected by either apamin/charybdotoxin, nifedipine, BayK 8644 or removal of external Ca2+. The short duration (approximately 100 ms), hyperpolarization-activated, postspike depolarizing afterpotentials (DAP), were significantly reduced by nifedipine. We propose that long duration ADPs underlie bursts and that short duration DAPs play a role in modulation of spike frequency. [ABSTRACT FROM AUTHOR]

Commercially farmed European flounder (Platichthys flesus) were treated with benzo[a]pyrene or cadmium under controlled laboratory conditions. In addition, animals were collected from the polluted Tyne and relatively clean Alde estuaries in the UK. Potential differentially expressed genes were isolated from liver tissue by means of suppressive, subtractive hybridisation. A total of 284 different mRNAs were identified as having the potential for being differentially expressed, of which 84 could be tentatively identified by homology with database sequences. These clones will form part of a flounder cDNA array for validation and future applications, with full identification being pursued for those expressed sequence tags for which differential expression can be confirmed by array technology. [Copyright &y& Elsevier]

The induction of hepatic cytochrome P450 1A (CYP 1A) is an important step in the response to contaminants such as polycyclic aromatic hydrocarbons (PAHs), and has been used as a biomarker of exposure in fish. Several consensus response elements have been identified, including eight potential xenobiotic response elements (XREs) in the promoter region of the European flounder cytochrome P450 1A gene. However not all of these sequences are necessarily active. To help elucidate the molecular regulation of this important gene, site directed mutagenesis and dual-luciferase reporter gene assays were employed to characterize the consensus transcription factor binding sites of the CYP 1A 5′ flanking region.Mutation of response elements situated -1103, -859, -709 and -172 bases upstream of the transcription start site reduced the induction to 2.75, 1.51, 3.25 and 3.05 fold, respectively, compared with the full-length promoter (4.0-fold induction) on exposure to the PAH 3-methylcholanthrene (3MC) (1.0 μM). These results indicate that four out of eight different XREs are functional in the control of CYP 1A in the flounder. The activity of these response elements adds to the evidence for considerable diversity in vertebrate CYP 1A regulation. [Copyright &y& Elsevier]

Vitellogenin (VTG) plasma concentrations were measured in flounder (Platicthys flesus L.) caught in two neighbouring UK estuaries between autumn 1997 and summer 2000. As found in previous studies, male and immature female flounder that were caught in the Mersey, but not in the Dee, had abnormally elevated concentrations of VTG in their plasma—evidence of the presence of natural and synthetic estrogens and/or xenoestrogens in the Mersey. The present study reveals a marked seasonal cycle in VTG concentrations in males. In late spring and summer, male flounder in the Mersey have relatively low concentrations of VTG (μg mL-1 range). However, in mid-winter, mean concentrations were as high as 5 mg mL-1. There is also evidence that there was a lessening in the degree of estrogenic endocrine disruption between 1996–1997 (the period of the initial studies on the Mersey) and 1998–2000 (the period covered by the present study). Only 0.5% of the fish contained ovotestes (intersex condition). [Copyright &y& Elsevier]

Hepatic microsomes of red mullet (Mullus barbatus) and flounder (Platichthys flesus) were preincubated in the presence of a concentration range of the antifouling agent tributyltin (TBT) chloride, and the interactions of TBT with cytochrome P450 and uridine diphosphate-glucuronyl transferase systems were investigated. The enzyme systems were examined in terms of cytochrome P4501A (CYP1A)-catalyzed 7-ethoxyresorufin O-deethylase (EROD) activity and benzo[a]pyrene (BaP) metabolism and in terms of glucuronidation of testosterone and 17β-estradiol, respectively. Ethoxyresorufin O-deethylase and BaP hydroxylase (BPH) activities of both fish species were progressively inhibited by increasing concentrations of TBT, and the effects were more pronounced for EROD than for BPH (maximal inhibition at 100 µM TBT for EROD and 250-500 µM TBT for BPH). Hydroxylated metabolites of BaP (3-hydroxy-, 7,8-dihydrodiol, and 9,10-dihydrodiol), representing 95% of the total metabolites formed, were reduced up to 75 % in the presence of 100 to 500 µM TBT, whereas the formation of other metabolites was less affected. This may alter BaP toxicity and carcinogenicity. Overall, the results were consistent with a specific inhibitory effect of TBT on CYP1A in the two fish species. Additionally, the conjugation of testosterone was significantly inhibited (20%) at low TBT doses (5 µM), with no effect on the glucuronidation of estradiol. [ABSTRACT FROM AUTHOR]

Plasma vitellogenin (VTG) concentrations and the presence of the ovo-testis (intersex) condition have been recorded in male flounder (Platichthys flesus) captured from several United Kingdom (UK) estuaries since 1996 as part of the endocrine disruption in the Marine Environment (EDMAR) project and earlier programs. It has been confirmed that plasma VTG concentrations in male flounder have remained elevated in several UK estuaries (e.g., Tees, Mersey, and Tyne) throughout the period covered by this study. However, the time-series data indicate that plasma VTG, a measure of environmental estrogen contamination, has decreased in fish captured from several estuaries, especially those of the Tyne and Mersey. Shorter time-series data sets from the Forth and Clyde estuaries also suggest a decrease in estrogen contamination at these sites. Trends associated with specific point sources of estrogenic contamination show site-specific patterns. For instance, plasma VTG levels in male flounder captured near the Howdon sewage treatment outfall (Tyne) have shown a steady decline to near baseline levels in 2001, while the plasma of male fish captured at a site adjacent to the Dabholm Gut discharge in the Tees estuary have shown little evidence of a sustained decline. The occurrence of the intersex condition was detected at a low but consistent prevalence through the study period, with the majority of cases recorded in fish captured from the Tyne and Mersey estuaries. The data set does not allow conclusions to be drawn about any temporal trends associated with this condition. The significance of the findings and possible mitigating influences are discussed in terms of the impacts on wild fish and the role of effluent treatment in reducing these. [ABSTRACT FROM AUTHOR]