PCR-based gut content analysis of insect predators: using ribosomal ITS-1 fragments from prey to estimate predation frequency

We used polymerase chain reaction to determine whether Ostrinia nubilalis (Hubner) (Lepidoptera: Crambidae) DNA was present in the guts of larvae and adult males and females of the generalist predator Coleomegilla maculata De Geer (Coleoptera: Coccinellidae). The predators were fed Ostrinia nubilalis egg masses and allowed to digest at either 20 degreesC or 27 degreesC for time spans ranging from 0 to 12 h. Four primer pairs, specific for 0. nubilalis were developed, using a nuclear ribosomal RNA sequence including part of the 18S gene, the complete internal transcribed spacer (ITS-1) region and part of the 5.8S gene. These primers amplified four sequences that were 492,369,256 and 150 base pairs long. We found a significant negative effect of time since feeding on the number of bands that could be detected. The shortest fragment was detected for the longest time after feeding (up to 12 h). We found no effect of predator weight, sex, developmental stage, or meal size on the time course over which bands of varying lengths could be detected.