Horse mackerel egg identification using DNA methodology

Abstract

Abstract Egg identification in plankton samples is usually needed for purposes of stock assessment. Until recently, only morphological characters were used for identifying the eggs of different fish species. Late developmental stages are easily distinguishable due to pigmentation as well as egg and oil globule size range. However, for early stages, these characteristics are difficult to be discriminated and may overlap with other species. European horse mackerel species ( Trachurus trachurus , T. mediterraneus , T. picturatus ) overlap significantly in their spawning areas in some European waters. Because of the fact that their eggs are morphologically similar, genetic methodologies are needed to identify eggs and larvae to species correctly. In the present study, formalin‐ and ethanol‐preserved eggs were tested to estimate the efficacy of genetic methodologies for species identification of eggs when different preservatives are used. A 370‐bp fragment of cytochrome b was successfully amplified followed by restriction fragment analysis with two restriction enzymes aiming to identify the eggs to species. Horse mackerel egg identification was accomplished with the maximum success in ethanol‐preserved eggs. However, it seems that various preservatives had different effects on the DNA quality of eggs as genetic identification was less successful in formalin‐preserved eggs. Preserving in ethanol a part of the eggs obtained in plankton surveys is suggested for purposes of accurate genetic identification, even if their morphological features are distorted in time.