2 research outputs found
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Species analysis of copepod nauplii in Florida Bay using molecular techniques
Copepod nauplii and their ecological roles within their respective communities are frequently overlooked because they are undersampled and difficult to identify. Morphological characters are often insufficient to differentiate between genera much less species. A rapid molecular identification assay was developed to differentiate species of copepod nauplii based on hybridization to species-specific oligonucleotide probes. The assay was successfully used to describe the species composition and abundance of the naupliar assemblage at four stations in Florida Bay through bi-monthly sampling over the course of one year (September 2001 to July 2002). Regional and seasonal variations in species composition and abundance of the naupliar assemblage were investigated with respect to the responses of different species to changes in environmental conditions. Differences between the species composition of the naupliar assemblage and the species composition of the adult copepod assemblage were detected. The naupliar assemblage was found to be higher in species richness, with many of the species only recognized by a DNA sequence (unnamed species). Phylogenetic analysis was performed in an effort to identify the unnamed species, many of which are likely to be benthic or parasitic as adults and therefore, were not found in the adult planktonic assemblage. This study presents evidence that all developmental stages should be included in copepod community analyses to provide a comprehensive understanding of the contribution of the community to the overall ecosystem
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Rapid identification of adult and naupliar stages of copepods using DNA hybridization methodology
Larval stages of common marine invertebrates and their ecological roles within their respective communities are frequently ignored because they are hard to identify. Morphological characters are often insufficient to differentiate between genera, much less species. To overcome the obstacles associated with species identification of copepod larvae, we developed a microtiter plate-based hybridization assay. Species-specific probes based on rDNA sequences were bound to microplates and used to capture target DNA. A novel method of linking the probes to the plate with poly-T tail ensured the probes were positioned above the plate surface and available for hybridization; this significantly increased the sensitivity of the assay. Target DNA extracted from individual copepods was amplified with biotin-labeled primers. The labeled target DNA bound to the probe specific for that species and produced a colorimetric change in the assay. The assay can be rapidly performed on freshly caught or ethanol preserved samples and the results visually interpreted