Primers to block the amplification of symbiotic apostome ciliate 18S rRNA gene in a PCR-based copepod diet study

Pelagic copepods play an important role in the marine food web. However, a full understanding of the ecological status of this zooplankton group depends on the careful study of their natural diets. In previous PCR-based copepod diet studies, we found many apostome ciliates that live symbiotically under the exoskeleton of the copepods, and their sequences were often over-represented in the 18S rRNA gene （18S rDNA） libraries. As a first step to address this issue, we designed three apostome ciliate 18S rDNA blocking primers, and tested their blocking efficiency against apostome ciliate 18S rDNA under various PCR conditions. Using a semi-quantitative PCR method, we optimized the conditions to efficiently amplify the 18S rDNA of the prey while simultaneously excluding the symbiotic apostome ciliates. This technique will facilitate PCR-based diet studies of copepods and other zooplankton in their natural environments.

In situ diet of the copepod Calanus sinicus in coastal waters of the South Yellow Sea and the Bohai Sea

Copepods are a key trophic link between primary producers and predatory animals at higher trophic levels in the marine ecosystem. Knowledge of the in situ composition of the copepod diet is critical for the accurate evaluation of trophic relationships and energy transfer in marine food webs. In this study, we applied a PCR-based cloning technique developed previously to investigate the in situ diet of Calanus sinicus, an ecologically important largesized calanoid copepod that dominates in the shelf waters around China, Japan and Korea. Analyses of the 18S rDNA sequences obtained from the copepod diet revealed the diverse food composition of C. sinicus from two stations （Y19 in the South Yellow Sea and B49 in the Bohai Sea）. A total of 43 operational taxonomic units （OTUs） were detected, which belonged to 13 diverse lineages： Bacillariophyta, Dinoflagellata, Dictyochophyceae, Chrysophyta, Katablepharidophyta, Pelagophyceae, Apusozoa, Hydrozoa, Ctenophora, Echinodermata, Tunicata, Chaetognatha and marine fungi. The results indicate that during an algae bloom, C. sinicus can graze on the bloom causative species. When the abundance of phytoplankton in ambient water is relatively low, C. sinicus can choose eggs, larvae, or organic particles/detritus of various metazoans, especially hydrozoans and ctenophores, as alternative food sources. Our result suggests that C. sinicus is an omnivorous species, and its prey choice may depend on the food availability in the ambient waters.