Preservation of Field-Collected Mosquito Blood Meals

Lawrence E Reeves; Nathan D Burkett-Cadena

doi:10.1101/pdb.prot108290

Preservation of Field-Collected Mosquito Blood Meals

Cold Spring Harb Protoc. 2023 Jul 17. doi: 10.1101/pdb.prot108290. Online ahead of print.

Authors

Lawrence E Reeves¹, Nathan D Burkett-Cadena²

Affiliations

¹ University of Florida, Institute of Food and Agricultural Sciences, Department of Entomology and Nematology, Florida Medical Entomology Laboratory, Vero Beach, Florida 32962, USA lereeves@ufl.edu.
² University of Florida, Institute of Food and Agricultural Sciences, Department of Entomology and Nematology, Florida Medical Entomology Laboratory, Vero Beach, Florida 32962, USA.

PMID: 37460150
DOI: 10.1101/pdb.prot108290

Abstract

All PCR- and DNA-based blood meal analyses require host DNA from a mosquito blood meal to be effectively preserved between the time when the specimen is collected and the extraction of DNA. As soon as a mosquito ingests blood from a host animal, digestion of host cells and cellular components within the blood meal by enzymes in the mosquito midgut begins to degrade the host DNA templates that are the targets of polymerase chain reaction (PCR) amplification. Without effective preservation, host DNA is typically undetectable by PCR 48 h after feeding, because of digestion. Preservation methods for mosquito blood meals vary in their efficacy, and the logistics of fieldwork can limit the options for preservation of blood meals and maintenance of the integrity of host DNA. This protocol describes a method of blood meal preservation that is effective, convenient, and amenable to fieldwork in remote locations where cryopreservation at -20°C or -80°C may not be feasible. It uses a Flinders Technology Associates (FTA) card, which is a chemically treated card that lyses cells and allows nucleic acids to be preserved. This method is also expected to preserve the DNA or RNA of pathogens present within the engorged mosquito abdomen, including RNA viruses.