Sampling design optimization for EROD measurements in fish

Activity of 7-ethoxyresorufin-O-deethylase (EROD) in fish is certainly the best-studied biomarker of exposure applied in the field to evaluate biological effects of contamination in the marine environment. Since 1991, a feasibility study for a monitoring network using this biomarker of exposure has been conducted along French coasts. Using data obtained during several cruises, this study aims to determine the number of fish required to detect a given difference between 2 mean EROD activities, i.e. to achieve an a priori fixed statistical power (l-beta) given significance level (alpha), variance estimations and projected ratio of unequal sample sizes (k). Mean EROD activity and standard error were estimated at each of 82 sampling stations. The inter-individual variance component was dominant in estimating the variance of mean EROD activity. Influences of alpha, beta, k and variability on sample sizes are illustrated and discussed in terms of costs. In particular, sample sizes do not have to be equal, especially if such a requirement would lead to a significant cost in sampling extra material. Finally, the feasibility of longterm monitoring is discussed.

Effect of temperature, food availability, and estradiol injection on gametogenesis and gender in the pearl oyster Pinctada margaritifera

The black-lip pearl oyster Pinctada margaritifera is a protandrous hermaphrodite species. Its economic value has led to the development of controlled hatchery reproduction techniques, although many aspects remain to be optimized. In order to understand reproductive mechanisms and their controlling factors, two independent experiments were designed to test hypotheses of gametogenesis and sex ratio control by environmental and hormonal factors. In one, pearl oysters were exposed under controlled conditions at different combinations of temperature (24 and 28°C) and food level (10,000 and 40,000 cells mL−1); whereas in the other, pearl oysters were conditioned under natural conditions into the lagoon and subjected to successive 17β-estradiol injections (100 μg per injection). Gametogenesis and sex ratio were assessed by histology for each treatment. In parallel, mRNA expressions of nine marker genes of the sexual pathway (pmarg-foxl2, pmarg-c43476, pmarg-c45042, pmarg-c19309, pmarg-c54338, pmarg-vit6, pmarg-zglp1, pmarg-dmrt, and pmarg-fem1-like) were investigated. Maximum maturation was observed in the treatment combining the highest temperature (28°C) and the highest microalgae concentration (40,000 cells mL−1), where the female sex tended to be maintained. Injection of 17β-estradiol induced a significant increase of undetermined stage proportion 2 weeks after the final injection. These results suggest that gametogenesis and gender in adult pearl oysters can be controlled by environmental factors and estrogens. While there were no significant effects on relative gene expression, the 3-gene-pair expression ratio model of the sexual pathway of P. margaritifera, suggest a probable dominance of genetic sex determinism without excluding a mixed sex determination mode (genetic + environmental)