Development of an acute toxicity test with the tropical marine amphipod Parhyale hawaiensis

Abstract

There is a lack of suitable tropical marine species for ecotoxicity tests. An attractive model organism for ecotoxicology is the marine amphipod Parhyale hawaiensis, which is already a model for genetic and developmental studies. This species is widespread, can tolerate changes in salinity, is easy to handle and is representative of circumtropical regions. The aim of this work was to describe standardized procedures for laboratory husbandry, define conditions for acute toxicity tests, and to provide acute toxicity test results for some reference toxicants. Culturing conditions for the organism in the laboratory were established in reconstituted seawater (30 ± 2 salinity), 24 ± 2 °C, photoperiod 12/12 h light/dark. Acute toxicity test procedures were developed for 96 h-exposure time, and organisms at ages <7 days. The miniaturized version of the test, based on 96-well microplates and 200 µL of exposure media provided consistent results compared to larger exposure volumes (80-mL vials protocol). Acute toxicity of Ag, Cd, Cu, Zn and ammonia determined for P. hawaiensis were consistent to previous results for other marine amphipods. We conclude that P. hawaiensis can be successfully cultured in standardized conditions and be effectively used in acute toxicity testing. Further development and use of this model will enable standardized and reproducible ecotoxicology investigations in understudied and vulnerable tropical marine ecosystems.

Toxicity responses for marine invertebrate species of brazilian occurrence

“…Culture and testing conditions for P. hawaiensis followed the protocol established by Artal et al (2018). Reconstituted seawater was used in the cultures and prepared with marine salt (Red Sea Salt, Red Sea®) dissolved in deionized water to achieve the desired salinity.…”

“…Additional cultures were maintained at salinity 5 for specific experiments and were maintained as described above. According to Artal et al (2018), two protocols of toxicity tests presented comparable results, one used vials of containing 25 mL of test solution and another using microplates with 200 µL of test solution in each well (miniaturized conditions). In this work we reported data with both protocols.…”

“…Silver nominal concentrations were 0.025, 0.05, 0.08, 0.1, 0.2, 0.25 mg L -1 for salinity 5, and 0.001, 0.01, 0,1, 0.25, 0.5, 1, 2, 4, 8, 10 mg L -1 for salinity 30. Zinc, cadmium and copper tests were conducted using a miniaturized testing approach develop by Artal et al (2018). Neonates were placed in 96-well microplates, with 32 replicas per concentration, 1 organism per well, in 200 µL of testing solution.…”

“…To address the urgent need to expand the available tropical marine species for use in toxicity testing, our laboratory has implemented culture conditions and developed toxicity tests for Parhyale hawaiensis (Artal et al, 2018), a deposit-feeding amphipod with circumtropical distribution and inhabits intertidal, mangrove and shallow waters of the Brazilian coast. P. hawaiensis was chosen because it is well established as a model in developmental and evolutionary studies (Kao et al, 2016) and recently successfully used in ecotoxicity tests (Artal et al, 2018). Another tropical and representative species is Nitocra sp., a harpacticoid copepod that inhabits region between mangrove swamps, and its culture was implemented in 1998 from organisms collected on sediment surface from intertidal zone of Spartina sp.…”

Viability of the marine amphipod Parhyale hawaiensis growth rate as an endpoint for chronic ecotoxicological tests

“…Parhyale hawaiensis has become a model for evolutionary and developmental biology research, but its potential for chronic toxicity assessment still unclear 1 . Thus, the aim of this study was to look for the minimum time interval that is possible to observe statistically significant growth to perform chronic testing, based on growth rate data to verify the application of this parameter as sublethal endpoint in chronic toxicity tests.…”