Identification of transcriptional effects of ethynyl oestradiol in male plaice (Pleuronectes platessa) by suppression subtractive hybridisation and a nylon macroarray

Suppression subtractive hybridisation (SSH) was used to generate cDNA libraries representing genes differentially expressed in response to ethynyl oestradiol (EE2) exposure in liver from male plaice (Pleuronectes platessa) previously analysed for vitellogenin (VTG) induction. Characterisation of the cDNA clones identified many as VTG (2 genes) and zona radiata proteins (ZRP) (3 genes), but 40 encoded other proteins, with more than half cryptic. Further analysis identified 85 non-redundant clones suitable for array on nylon membrane. Radiolabelled cDNAs were prepared from hepatic mRNA from EE2 treated plaice (0 and 21 days) and hybridised with the arrayed clones. Analysis of the data showed that 11/17 novel, 21/22 VTG, 13/14 ZRP, 2/2 liver aspartic proteinase (LAP) and 8/10 other mRNAs were up-regulated by EE2 exposure.     

Petroleum contamination and biochemical alterations in oysters (Crassostrea gigas) and plaice (Pleuronectes platessa) from bays impacted by the Amoco Cadiz crude oil spill

Following the Amoco Cadiz crude oil spill in March 1978, marine animals in coastal and estuarine environments along the north shore of Brittany, France became heavily contaminated with crude oil. The resident benthic fauna in the oil-impacted area which survived the spill were severely stressed by the pollution. The objectives of our investigation were to document long-term trends in petroleum hydrocarbon contamination and sublethal stress in oysters and plaice from the estuaries, Aber Benoit and Aber Wrac'h. Oysters in the estuaries accumulated aliphatic and aromatic petroleum hydrocarbons to high levels and remained heavily contaminated for the duration of the 27 month investigation. Plaice, on the other hand, retained only low concentrations of petroleum hydrocarbons in muscle and liver tissue. Nevertheless, the oysters exhibited very few histopathologic and biochemical responses to the oil, whereas the plaice from the estuaries exhibited a variety of sublethal histopathologic and biochemical alterations.     

The effect of salinity on growth and weight loss of juvenile plaice (Pleuronectes platessa, L): An experimental test

Previous population estimates of the 0+ plaice (Pleuronectes platessa L.) in the Firth of Forth, east central Scotland, did not take account of the Forth estuary west of the Forth bridges. Previous work found plaice in the estuary grew as fast as, or faster than, the outer firth plaice. It was hypothesised that salinity may affect growth rates of early 0+ plaice. This hypothesis was tested in a laboratory experiment, by exposing juvenile plaice to three different, but naturally — experienced by the juveniles, salinities; 25, 30 and 35. Plaice fed a minimum ration did not grow in length. Mean weight decreased at all three salinities, however, the lowest weight loss was found at the lowest salinity (25) and the highest weight loss was found at the highest salinity (35). The minimum feeding ration was halted and plaice were then fed ad libitum. Consumption rates were not significantly different during the ad libitum feeding, while significant differences in mean weight change were found between the highest and lowest salinities.     

The distribution and population dynamics of O-group plaice (Pleuronectes platessa L.) on nursery grounds in the Firth of Forth

Midsummer (1 August) population estimates of about 2 million O-group plaice (Pleuronectes platessa L.) were derived for sandy bays around the Firth of Forth in 1979–1980. This is an order of magnitude less than similar estimates made for the Clyde Sea Area in 1973–1974. Autumn population estimates of 0·4–1·0 million fish were comparable to estimates by the Ministry of Agriculture, Fisheries and Food for the area between the Scottish border and Flamborough Head (2·3 million for 1970 and 1973) which represented 4·8% (1973) to 5·3% (1970) of the total number of O-group fish on the English east coast.Largo Bay was the most important nursery area holding 25% of the total population. It is particularly well situated to receive newly metamorphosed plaice carried in water currents along the north side of the Forth from the spawning ground off Fife Ness. Plaice in the Forth are mainly distributed on fine to medium sandy beaches (186–480 μm), the mean number per haul in midsummer (D) being correlated with the median diameter (m.d. in μm) of the low water sediments by the equation: D=−45·7666+0·2327 m.d. (n=11,r=0·68,P<0·02 but>0·01).The shallow inshore water in sandy bays in the outer Firth was well mixed and more marine than estuarine (27·7–35·0‰). The correlation coefficient between fish density and water temperature was low, while that with salinity (S‰) was: D=6·1618+0·2238S (n=23,r=0·62,P<0·005).Regression analysis demonstrated that the relationship between the instantaneous mortality rate (Z) and the initial population density (D_p) was: Z×100=0·7480+0·0546d_p (n=12,r=0·87,P<0·001).The mean mortality rate for the O-group plaice in the Forth nursery areas was 53% month⁻¹.     

Modelling the spatial distribution of plaice (Pleuronectes platessa), sole (Solea solea) and thornback ray (Raja clavata) in UK waters for marine management and planning

Species distribution maps are needed for ecosystem-based marine management including the development of marine spatial plans. If such maps are based on predictive models then modelling procedures should aim to maximise validation success, and any uncertainty in the predictions needs to be made explicit. We developed a predictive modelling approach to produce robust maps of the distributions of selected marine species at a regional scale. We used 14 years of survey data to map the distributions of plaice, sole and thornback ray in three hydrographic regions comprising parts of the Irish Sea, Celtic Sea and the English Channel with the help of the hybrid technique regression kriging, which combines regression models with geostatistical tools. For each species–region combination we constructed logistic Generalized Linear Models (GLMs) based on presence–absence data using the environmental variables: depth, bottom temperature, bed shear stress and sediment type, as predictors. We selected GLMs using the mean squared error of prediction (MSEP) estimated by cross-validation then conducted a geostatistical analysis of the residuals to incorporate spatial structure in the predictions. In general, we found that species occurrence was positively related to shallow areas, a bed shear stress of between 0 and 1.5 N/m², and the presence of sandy sediment. Predicted species occurrence probabilities were in good agreement with survey observations. This modelling framework selects environmental models based on predictive ability and considers the effect of spatial autocorrelation on predictions, together with the simultaneous presentation of observations, associated uncertainties, and predictions. The potential benefit of these distribution maps to marine management and planning is discussed.     

o,p′-DDT induction of vitellogenesis and its inhibition by tamoxifen in Nile tilapia (Oreochromis niloticus)

In order to investigate the mechanism by which o,p′-DDT disrupts endocrine functioning of Nile tilapia in vivo, the estrogenicity of o,p′-DDT was investigated in conjunction with 17β-estradiol (E₂) and tamoxifen. Mature, male tilapia were treated intraperitoneally with o,p′-DDT (60 mg/kg, one dose) or E₂ (5 mg/kg, four doses) in the presence or absence of tamoxifen (5 mg/kg, six doses) for 12 days and then plasma vitellogenin (Vtg) (measured as alkaline-labile phosphorous), E₂, and testosterone (T) were measured. Vtg levels were increased dramatically by E₂ (1744±171 μg/ml) and moderately by o,p′-DDT (82±15 μg/ml) compared with controls (23±3.5 μg/ml). Tamoxifen alone had no effect on Vtg production, but inhibited both E₂ and o,p′-DDT stimulated vitellogenesis. T levels were reduced with E₂ administration (1688±383 pg/ml) and declined further with the combined treatment of E₂ and tamoxifen (281±70 pg/ml), compared with controls (6558±1438 pg/ml). Tamoxifen or o,p′-DDT alone did not affect T levels, but their combined treatment did (2069±647 pg/ml). The results of this study suggest that o,p′-DDT is weakly estrogenic in male tilapia, and that this activity may be mediated through the estrogen receptor.     

Biochemical and histological responses in mussel (Mytilus edulis) exposed to North Sea oil and to a mixture of North Sea oil and alkylphenols

Several environmental chemicals are suspected to be responsible for adverse health effects on the reproductive system in various organisms. During this work, environmentally relevant concentrations of North Sea oil were used alone or in combination with alkylphenols and additional PAH to study the effect on vitellogenin-like protein expression and gonadal development in mussels. North Sea oil (0.5 ppm) induced the expression of phospho-proteins in both sexes indicating that some compounds are oestrogen-mimics. This induction was not seen in samples dosed with the mixture but signs of toxic effects were observed in the gonads. Indeed, numerous degenerating ovarian follicles in females and foci, similar to vertebrate melanomacrophage centres, were observed in testes.     

Use of suppressive subtractive hybridization and cDNA arrays to discover patterns of altered gene expression in the liver of dihydrotestosterone and 11-ketotestosterone exposed adult male largemouth bass (Micropterus salmoides)

In this study male largemouth bass (LMB) were exposed to the naturally occurring androgens, dihydrotestosterone (DHT) or 11-ketotestosterone (11-KT) in order to identify genes that are differentially regulated by these steroid hormones. Using subtractive hybridization on livers of fish treated with DHT against vehicle control, many novel LMB genes were cloned. These genes were added to our gene library and arrayed. Six genes were up-regulated and five were down-regulated by both androgens. But, each androgen also regulated specific genes. One gene that was identified as a potential androgen marker was spermidine-spermine-N¹-acetyltransferase that was up-regulated by both androgens. Determining which genes are responsive to natural androgens will help to identify biochemical pathways that are impacted.