Selenium poisoning of fish by coal ash wastewater in Herrington Lake, Kentucky.

Selenium pollution from the E.W. Brown Electric Generating Station was investigated in Herrington Lake, KY. Coal ash wastewater is discharged as surface water overflow from ash disposal ponds into the lake via a National Pollutant Discharge Elimination System permit issued by the Kentucky Division of Water, but the permit does not restrict or limit the amount of selenium released. Unpermitted discharges occur from seeps and drainage through leaks in ash pond dams. Together, these discharges have resulted in selenium concentrations in water, sediment, benthic macroinvertebrates, and fish that are 2-9 times the level that is toxic for fish reproduction and survival. A large proportion (12.2%, or 25 times background) of juvenile largemouth bass (Micropterus salmoides, the only species examined) exhibited spinal and/or craniofacial malformations that are consistent with selenium poisoning. Teratogenic Deformity Index values indicated a 3.05% population-level impact on the bass fishery, with total selenium-induced mortality (including pre-swimup mortality) estimated to be in excess of 25% per year. These findings confirm that coal ash discharges into Herrington Lake are contributing selenium to the Lake that is poisoning fish.

Saddleback syndrome in European sea bass Dicentrarchus labrax (Linnaeus, 1758): anatomy, ontogeny and correlation with lateral-line, anal and pelvic fin abnormalities.

This study focused on the anatomy and ontogeny of saddleback syndrome (SBS) in reared European sea bass. The abnormality was detected at an unusually high frequency (12-94%) during a routine quality control in a commercial hatchery. Anatomically the abnormality was mainly characterized by the loss of 1-5 hard spines and severe abnormalities of the proximal pterygiophores (anterior dorsal fin), size reduction of a few lepidotrichia, missing lepidotrichia and/or lepidotrichia of poor ossification (posterior dorsal fin). SBS was significantly correlated with abnormalities of the anal and pelvic fins in all the examined populations. Moreover, in juvenile fish, SBS was significantly correlated with an abnormal lack of sectors of the lateral line. The examination of early larval samples revealed that SBS was ontogenetically associated with severe abnormalities of the primordial marginal finfold, which developed at the flexion stage (ca 8.5-11.0 mm SL). Histologically, these abnormalities were associated with extensive epidermal erosion. The results are discussed in respect of the critical ontogenetic period and the possible causative factors of SBS in European sea bass. It is suggested that causative factors acted during a wide ontogenetic period including flexion and metamorphosis phases.

Effect of an experimental oil spill on vertebral bone tissue quality in European sea bass (Dicentrarchus labrax L.).

In order to identify biomarkers of oil pollution in fish we tested the effects of an experimental Light Cycle Oil (LCO) exposure on vertebral bone of sea bass, Dicentrarchus labrax L. A total of 60 adult fish were acclimated for fifteen days, then twenty were collected as controls (Day 0) while 40 were exposed to a soluble fraction of LCO (1136 ng L(-1) of ten Polycyclic Aromatic Hydrocarbons, PAHs) for seven days. Twenty of them were sampled at the end of the exposure period and the twenty last after a recovery period of fourteen days in clean seawater. Vertebral abnormalities were counted and bone mineralization, total bone area and bone density profiles were established for several post-cranial and caudal vertebrae. In sea bass, seven days of LCO exposure did not affect the frequency and severity of the vertebral abnormalities. No significant differences were observed in bone density and bone repartition (parameters of bone area profiles) between unexposed (Day 0), exposed (D7) and decontaminated (D21) fish. In contrast, bone mineralization of the vertebrae decreased in contaminated sea bass, but in a reversible way, which confirms a previous study in trout showing that this parameter is an early stress indicator. Our results suggest that vertebral bone mineralization could be used as a biomarker of PAH pollution in sea bass. It would be interesting to check this new biomarker in other teleost species exposed to various xenobiotics.

Development of an oligo DNA microarray for the European sea bass and its application to expression profiling of jaw deformity.

BACKGROUND: The European sea bass (Dicentrarchus labrax) is a marine fish of great importance for fisheries and aquaculture. Functional genomics offers the possibility to discover the molecular mechanisms underlying productive traits in farmed fish, and a step towards the application of marker assisted selection methods in this species. To this end, we report here on the development of an oligo DNA microarray for D. labrax. RESULTS: A database consisting of 19,048 unique transcripts was constructed, of which 12,008 (63%) could be annotated by similarity and 4,692 received a GO functional annotation. Two non-overlapping 60mer probes were designed for each unique transcript and in-situ synthesized on glass slides using Agilent SurePrint technology. Probe design was positively completed for 19,035 target clusters; the oligo microarray was then applied to profile gene expression in mandibles and whole-heads of fish affected by prognathism, a skeletal malformation that strongly affects sea bass production. Statistical analysis identified 242 transcripts that are significantly down-regulated in deformed individuals compared to normal fish, with a significant enrichment in genes related to nervous system development and functioning. A set of genes spanning a wide dynamic range in gene expression level were selected for quantitative RT-PCR validation. Fold change correlation between microarray and qPCR data was always significant. CONCLUSIONS: The microarray platform developed for the European sea bass has a high level of flexibility, reliability, and reproducibility. Despite the well known limitations in achieving a proper functional annotation in non-model species, sufficient information was obtained to identify biological processes that are significantly enriched among differentially expressed genes. New insights were obtained on putative mechanisms involved on mandibular prognathism, suggesting that bone/nervous system development might play a role in this phenomenon.

Biochemical and toxicopathic biomarkers assessed in smallmouth bass recovered from a polychlorinated biphenyl-contaminated river.

Smallmouth bass (Micropterus dolomieu) were collected to quantify the nature and prevalence of biomarker responses, including biochemical indices, toxicopathic lesions and general health indices, among fish collected from polychlorinated biphenyl (PCB)-contaminated and nearby uncontaminated reaches of the Kalamazoo River, Michigan, USA. Blood and tissue samples (gill, liver, spleen, head kidney, trunk kidney, thyroid and gonads) were collected and preserved at necropsy for biochemical and histological analyses. The body condition factor and liver somatic index were significantly lower in fish collected from the downstream, contaminated site. Plasma vitellogenin was not detected in male fish collected from either site. Liver ethoxyresorufin-O-deethylase activity and liver and spleen superoxide dismutase activity were significantly depressed in fish collected from the downstream site. Significant toxicopathic lesions such as glycogen depletion, enhanced macrophage aggregates, hepatic foci of cellular alteration (i.e. preneoplastic lesions) and neoplasia were also detected in the liver of fish collected from the downstream site. This study indicates that many of the biochemical and histopathological biomarker responses were associated with liver and body tissue PCB concentrations. Taken together, the biomarkers of exposure and effect strongly suggest that fish within the downstream site are adversely affected by PCBs and other chemical stressors.