Sample Collection
Collection of EW was carried out at the Bangkok Aquaculture Farm Company, Nakhon Sri Thammarat Province, Thailand. Female shrimp possessing mature ovaries (stage IV) were individually placed in a 500-L plastic tank. In order to exclude sperm factors in the EW, only shrimp that had not been inseminated were used in this study. The shrimp were held firmly over a 500-ml container and allowed to spawn their eggs into filtered seawater. After swirling gently to settle the eggs to the bottom, seawater without spawned eggs was collected and designated as EW. This EW was centrifuged (10 000 × g, 15 min, 4°C) to remove particulates, and kept at −80°C until use. Proteins in the EW were quantified with a Bradford's reagent