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

Débora P. Paula, David A. Andow

Research output: Contribution to journalArticlepeer-review

Abstract

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

Original languageEnglish (US)
Pages (from-to)1552-1561
Number of pages10
JournalMethods in Ecology and Evolution
Volume12
Issue number9
DOIs
StatePublished - May 17 2021

Bibliographical note

Funding Information:
We thank Dr. Valmir A. Costa (Instituto Biológico de São Paulo‐Brazil) and Professor José Roberto Pujol Luz (Universidade de Brasília‐Brazil) for taxonomic determinations, all the farmers that allowed the host sampling in 2017 and 2018 on their properties, and all undergraduate students, researchers and professors who participated in the host sampling in the field. We also thank Embrapa for the financial support by the grant number SEG 12.13.12.005.00.00. The authors confirm they have no conflict of interest.

Publisher Copyright:
© 2021 British Ecological Society

Keywords

  • Coccinellidae
  • Dinocampus coccinellae
  • Harmonia axyridis
  • Homalotylus
  • arthropods
  • melting curve analysis
  • qPCR

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