RT Journal Article SR Electronic T1 Novel Insights into Malaria Vector Surveillance in Madagascar Using a Quadrant Enabled Screen Trap (QUEST) and Bloodmeal Detection Assay for Regional Transmission (BLOODART) JF bioRxiv FD Cold Spring Harbor Laboratory SP 530758 DO 10.1101/530758 A1 Riley E. Tedrow A1 Tovonahary Rakotomanga A1 Thiery Nepomichene A1 Jocelyn Ratovonjato A1 Arséne C. Ratsimbasoa A1 Gavin J. Svenson A1 Peter A. Zimmerman YR 2019 UL http://biorxiv.org/content/early/2019/01/25/530758.abstract AB Background The Madagascar National Strategic Plan for Malaria Control 2018 (NSP) outlines malaria control pre-elimination strategies that include detailed goals for mosquito control. Primary surveillance protocols and mosquito control interventions focus on indoor vectors of malaria, while many potential vectors feed and rest outdoors. Here we describe the application of novel tools that advance our understanding of diversity, host choice, and Plasmodium infection in the Anopheline mosquitoes of the Western Highland Fringe of Madagascar.Methodology/Principal Findings We employed a novel outdoor trap, the QUadrant Enabled Screen Trap (QUEST), in conjunction with the recently developed multiplex BLOOdmeal Detection Assay for Regional Transmission (BLOODART). We captured a total of 1252 female Anopheles mosquitoes (10 species), all of which were subjected to BLOODART analysis. QUEST collection captured a heterogenous distribution of mosquito density, diversity, host choice, and Plasmodium infection. Concordance between Anopheles morphology and BLOODART species identifications ranged from 93-99%. Mosquito feeding behavior in this collection frequently exhibited multiple blood meal hosts (single host = 53.6%, two hosts = 42.1%, three hosts = 4.3%). The overall percentage of human positive bloodmeals increased between the December 2017 and the April 2018 timepoints (27% to 44%). Plasmodium positivity was found primarily in vectors considered to be of secondary importance, with an overall prevalence of 6%.Conclusions/Significance The QUEST was an efficient tool for sampling Anopheline mosquitoes. Vectors considered to be of secondary importance were commonly found with Plasmodium DNA in their abdomens, indicating a need to account for these species in routine surveillance efforts. Mosquitoes exhibited multiple blood feeding behavior within a gonotrophic cycle, with predominantly non-human hosts in the bloodmeal. Taken together, this complex feeding behavior could enhance the role of multiple Anopheline species in malaria transmission, possibly tempered by zoophilic feeding tendencies.Author Summary Malaria continues to be a significant threat to public health in Madagascar. Elimination of this disease is impeded by numerous factors, such as vector surveillance that does little to account for the potential role of secondary malaria vectors, which rest and feed outdoors. In this study, we designed a novel, low cost QUadrant Enabled Screen Trap (QUEST) to address the lack of traps for outdoor mosquitoes. We used this in conjunction with our novel BLOOdmeal Detection Assay for Regional Transmission (BLOODART) to assess mosquito feeding behavior in the Western Highland Fringe of Madagascar. Our analysis revealed significant variability in mosquito density, diversity, host choice, and Plasmodium infection across traps placed within and between two nearby villages at two timepoints; indicating a strong, small-scale spatial component to disease transmission that warrants further investigation. Many of the mosquitoes in this sample (46.4%) fed on two or three host species, indicating complex feeding behaviors that could influence malaria transmission. Further, Plasmodium DNA was detected in the abdomens of numerous vectors of supposed secondary importance, indicating a neglected parasite reservoir and an increased need to account for these species in routine surveillance efforts.