The impact of selected abiotic factors on Artemia hatching process through real-time observation of oxygen changes in a microfluidic platform.

Current studies on abiotic impacts on Artemia, a crustacean which is widely used in aquaculture, and ecotoxicology, often focus on endpoint analysis (e.g., hatching rates, survival). Here, we demonstrate that a mechanistic understanding can be obtained through measurement of oxygen consumption in real-time over an extended time period in a microfluidic platform. The platform enables high level control of the microenvironment and direct observation of morphological changes. As a demonstration, temperature and salinity are chosen to represent critical abiotic parameters that are also threatened by climate change. The hatching process of Artemia consists of four different stages: hydration, differentiation, emergence, and hatching. Different temperatures (20, 35, and 30 °C) and salinities (0, 25, 50, and 75 ppt) are shown to significantly alter the duration of hatching stages, metabolic rates, and hatchability. Specifically, the metabolic resumption of dormant Artemia cysts was significantly enhanced at higher temperatures and moderate salinity, however, the time needed for this resumption was only dependent on higher temperatures. Hatchability was inversely related to the duration of the differentiation stage of hatching, which persisted longer at lower temperatures and salinities. The current approach of investigation of metabolism and corresponding physical changes can be employed to study hatching processes of other aquatic species, even those with low metabolic rate.