Identification of the Distribution and Species Composition of the Anopheles crucians Cryptic Species Complex in Lowndes County, GA, USA, and the Prevalence of Dirofilaria immitis within the An. crucians Complex in Lowndes County, GA, USA

Anopheles crucians s.l. includes several understudied cryptic mosquito species. This species complex is comprised of six morphologically indistinguishable species. Female mosquitoes were obtained from 14 collection sites in Lowndes County, GA, USA during the 2014 trapping season. The internal transcribed spacer two (ITS2) gene was amplified in individual mosquitoes and used to identify them at the species level. Of the six species within the cryptic species complex, only three species (A, C, and D) were identified within Lowndes County, GA, USA. Sympatry was observed between the species at different collection sites, with four sites having all three species in sympatry and other sites having only two species in sympatry. Anopheles crucians s.l. has been implicated as a vector of Dirofilaria immitis, as well as other pathogens, but its competency has not been fully confirmed. The collected mosquitoes were screened for the presence of D. immitis using a polymerase chain reaction (PCR) protocol to amplify the 16S rRNA gene from dog heartworm within the host mosquitoes. The heads and thoraces were separated from the abdomens of the mosquitoes, and pools of each were screened for D. immitis DNA by PCR. Dirofilaria immits DNA was detected in a single pool of An. crucians abdomens. Future studies could be aimed toward screening the An. crucians cryptic species complex for competency in vectoring other pathogens and arboviruses. TABLE OF CONTENTS Chapter I: INTRODUCTION 1 | Materials and Methods 6 | Specimen Collection 6 | DNA Extraction 6 | PCR Amplification 7 | Results 7 | Discussion 8 | Chapter II: INTRODUCTION 11 | Materials and Methods 13 | Specimen Collection 13 | DNA Extraction 14 | PCR Amplification 15 | Statistical Analysis 15 | Results 16 | Discussion 16 | Chapter III: CONCLUSION 19 | APPENDIX A: Figures for Chapters 1-3 28 | APPENDIX B: Tables for Chapters 1-3 40 |Chambers, Eric W.Blackmore, Mark S.Gosnell, Donna L.M.S.Biolog