Evaluating food web interactions among microcrustaceans and insect in a tropical shallow lake using DNA-based protocol.

We developed species-specific primers of five microcrustacean preys, Ceriodaphnia richardi, Diaphanosoma cf. brevireme, Daphnia gessneri, Simocephalus serrulatus, Thermocyclops decipiens and Mesocyclops sp., to analyze food-web interactions involving their two insect predators Rheumatobates crassifemur and Martarega uruguayensis distributed in a tropical shallow lake. We designed internal primers of the COI gene (177-282 bp), and tested them, by means of PCR, for specificity and sensitivity. In our tests for specificity, all primers successfully amplified the DNA target but were species-specific failing to amplify the biomarker from any of the other species tested, even in a mixed DNA sample, including predators' DNA. In tests for sensitivity, primers successfully amplified zooplankton biomarkers from low concentration of DNA extractions and also from digestive tract of predators, even after many hours of ingestion. This technique provides a framework as an efficient tool for evaluation of food-web research in natural aquatic environments, where it is impossible to observe if predation occurs. Furthermore, this technique provides an effective solution for the identification of zooplankton species from the predator's digestive tract, where morphological identification alone is sometimes difficult because predators do not consume the prey but feeds using extra-oral digestion, such is the case of heteropterans.