Melting curve analysis for detection and identification of ghost parasitoids in host carcasses a month after host death

Incidence of parasitism is often underestimated because ‘ghost’ parasitoids (dead unemerged or those that have emerged leaving the host carcasses) are difficult to detect and identify. This study demonstrates use melting curve analysis (MCA) carcasses can identify DNA ghost even a month after death. The coccinellid hosts Cycloneda sanguinea, Eriopis connexa, Harmonia axyridis Hippodamia convergens were sampled from cole crops in 2017 2018 Midwest Brazil, reared observed daily for parasitoid emergence. Dead coccinellids held 30 days death before storage only with emergence analysed. Species-specific primers designed identification species during rearing: Dinocampus coccinellae, Homalotylus mirabilis, Ho. terminalis, Strongygaster triangulifera Phalacrotophora sp. temperatures (Tm) their amplicons used as positive controls MCA post-amplification qPCR. Detection was possible D. mirabilis limit detection (LOD) all <1 pg DNA, except Phalacroptopthora (LOD = 1.9 ng). Parasitoids detected 31 out 70 (44.3%) ranging 0% 71%, depending on species. Synthesis applications. work could be rapidly some (e.g. coccinellae sp.) up high sensitivity specificity. any field-collected and, species, it should lead more accurate estimates incidence parasitism. Our approach easily adapted applied other parasitoid–host interactions.