Biochemical Composition of Naupii Derived from Stored Non-Diapause and Diapause Copepod Eggs and the Biology of Diapausing Eggs

Mariculture of fish is needed to support our ever growing global population. Copepods are a natural and beneficial source of first feed for many marine fish species, but it can be difficult to hatch enough when the fish are ready to feed. Therefore, storage of copepod eggs for use at a later date increases the potential of nauplii to be used as a commercial food source. There are two types of eggs produced by copepods: diapause and non-diapause (or subitaneous). However, little is known about diapause or the effects and duration of storage, either under cold or warm temperature conditions, on the biochemical composition of nauplii. Little is also known about the development of embryos that enter diapause. Diapause requires a refractory period before development of the embryo continues. The refractory period can last for several months regardless of environmental conditions. I studied the effects of both cold storage and cold storage with the addition of antibiotics on non-diapause eggs of the copepod Acartia tonsa and duration of storage for diapause eggs produced by Centropages hamatus and compared those results to nauplii derived from non-diapause eggs. The organic components analyzed to determine if potential changes were occurring during storage were lipids, fatty acids, proteins, free amino acids, and carbohydrates as well as the percent hatch of the eggs and the dry weight of the resultant nauplii. To understand diapause, we utilized two stains, one to determine the number of nuclei present and another to determine intracellular pH of the diapausing eggs. Acartia tonsa eggs stored for up to15 days at 1°C did not indicate any change in the biochemical make-up of the resulting nauplii. The only change we observed was in the viability of the eggs, which decreased at a steady rate over time. The viability of the eggs quickly approached zero percent hatch beyond 15 days. The addition of the antibiotic oxytetracyclin at a 10% concentration did not change the naupliar biochemistry and did not increase viability over the storage time. Centropages hamatus eggs maintained a high level of viability over the course of 13 months of storage under anoxic conditions. The nauplii derived from the diapause eggs stored at 25°C had similar biochemical components regardless of the length of the storage period. My study indicates that storage of A. tonsa and C. hamatus may not affect the nutritional value of the nauplii for aquaculture purposes. We also determined that the embryos of C. hamatus stopped developing after ~7 cleavages. The diapausing embryos also maintained an intracellular pH similar to the surrounding water and acidified when beginning to develop. This is the first time the intracellular pH and only the third time the nuclei of a copepod diapausing egg has been determined. This information could allow future researchers to interrupt diapause and induce the eggs to hatch before the end of the refractory period.