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MB Sample ID: SA245787
| Local Sample ID: | bivalves_21C_C125_02 |
| Subject ID: | SU002545 |
| Subject Type: | Invertebrate |
| Subject Species: | Ruditapes philippinarum |
| Taxonomy ID: | 129788 |
| Height Or Height Range: | length: 3.81 ± 0.42 cm; and width: 3.06 ± 0.51 cm |
| Gender: | Male and female |
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Collection:
| Collection ID: | CO002538 |
| Collection Summary: | Ruditapes philippinarum clams were collected at the Ria de Aveiro, a shallow coastal system located on the Northwest Atlantic coast of Portugal. Individuals of similar size (length: 3.81 ± 0.42 cm; and width: 3.06 ± 0.51 cm) were selected. For depuration and acclimation to laboratory conditions, all clams were maintained in artificial seawater for 10 days (salinity: 30 ± 1, Tropic Marin® SEA SALT, from Tropic Marine Center), under continuous aeration, constant temperature (17 ± 1 ºC) and a natural photoperiod. Artificial seawater was renewed every 2-3 days and clams were fed every 2-3 days with Algamac Protein Plus (150,000 cells/animal/day) after the 3rd day. After depuration, the organisms were subjected to a chronic toxicity test for 28 days, consisting of exposure to five different EE2 concentrations (Sigma-Aldrich, purity ≥ 98%, MW = 296.40 g/mol, 1 mg/L stock solution in ultrapure water): 0 (control group), 5, 25, 125 and 625 ng/L. To assess the effects of a warming scenario on the impacts of EE2, the experiments were carried out at 17 ± 1 °C (control; mean temperature of sampling area during September: 16 - 19 °C) and at 21 ± 1 °C (worst-case climate change scenario, IPCC, 2021). The aquaria were placed in distinct climatic rooms for each temperature. To reach 21 °C, the temperature was raised by 2 °C, every 2–3 days. For each concentration level and temperature, 12 samples were considered: 4 individuals per aquarium and 3 aquaria per treatment. In each aquarium, a total of 3 L of artificial seawater (salinity: 30 ± 1), continuous aeration, and a natural photoperiod were used. The exposure medium for each condition was renewed weekly, after which EE2 concentration levels were re-established. Mortality was checked daily and found to be null. At the end of the 28-day exposure period, the clams were frozen in liquid nitrogen and stored at – 80 ºC. |
| Collection Protocol Filename: | Bivalves_Experimental_Procedure.docx |
| Sample Type: | Tissue |
| Storage Conditions: | -80℃ |
