Molecular Identification of Vertebrate and Hemoparasite DNA Within Mosquito Blood Meals From Eastern North Dakota

Abstract

To understand local transmission of vector-borne diseases, it is important to identify potential vectors, characterize their host feeding patterns, and determine if vector-borne pathogens are circulating within the region. This study simultaneously investigated these aspects of disease transmission by collecting engorged mosquitoes within two rural study sites in the central Red River Valley of North Dakota. Mosquitoes were identified, midguts were excised, and the blood was expelled from the midguts. DNA was extracted from blood meals and subjected to PCR and direct sequencing to identify the vertebrate origin of the blood. Using different primer sets, PCR was used to screen for two types of vector-borne pathogens, filarioid nematodes and hemosporidian parasites. White-tailed deer were the primary source of blood meals for the eight aedine mosquito species collected. None of the 288 deer-derived blood meals contained filarioid or hemosporidian DNA. In contrast, 18 of 32 Culex tarsalis and three of three Cx. pipiens blood meals contained avian blood, representing eight different species of birds. Of 24 avian-derived blood meals examined, 12 contained Plasmodium DNA, three of which also contained Leucocytozoon DNA (i.e., dual infection). Potential confounding effects resulting from parasite acquisition and development from previous blood meals (e.g., oocysts) were eliminated because host blood had been removed from the midguts prior to DNA extraction. Thus, specific parasite lineages/species could be unequivocally linked to specific vertebrate species. By combining mosquito identification with molecular techniques for identifying blood meal source and pathogens, a relatively small sample of engorged mosquitoes yielded important new information about mosquito feeding patterns and hemosporidia infections in birds. Thorough analyses of wild-caught engorged mosquitoes and other arthropods represent a powerful tool in understanding the local transmission of vector-borne and zoonotic diseases.

Host preferences in host‐seeking and blood‐fed mosquitoes in Switzerland

“…Analyses of the blood‐fed mosquitoes from the zoo and the field with different primer pairs resulted in the successful identification of blood hosts in 91.9% of specimens, which is higher than in previous studies (Townzen et al ., ; Lassen et al ., ; Tuten et al ., ; Mehus & Vaughan, ) in which single or multiple (as in the present study) PCR approaches were employed. The primers used in the present study (Kocher et al ., ; Ngo & Kramer, ; Townzen et al ., ) were not sensitive (i.e.…”

Metagenomic sequencing of dung beetle intestinal contents directly detects and identifies mammalian fauna

“…The contents of invertebrate intestines is becoming a widely-used source of environmental DNA (eDNA) in biodiversity sampling (Calvignac-Spencer et al 2013a). That fragments of vertebrate DNA can be successfully extracted, amplified, sequenced, and identified from invertebrate digestive tracts has already been demonstrated in several studies, including those using blood-feeding leeches (Schnell et al 2012) and mosquitos (Towzen, Brower & Judd 2008; Mehus & Vaughan 2013), and carrion-feeding flies (Calvignac-Spencer et al 2013b) as the source material.…”

Experimental logging alters the abundance and community composition of ovipositing mosquitoes in the southern Appalachians

“…Using a silvicultural experimental approach that applied two types of logging treatment (single-entry clearcut, and repeated-entry shelterwood harvesting) to otherwise similar forest plots, here we sought to understand how logging influences the abundance, diversity, and community composition of mosquitoes in southwest Virginia. As this assemblage includes the vectors of avian malaria (Kimura et al, 2010;Farajollahi et al, 2011), reptile haemogregarines (Smith et al, 1996;Harkness et al, 2010), filarial worms (Ledesma & Harrington, 2011;Mehus & Vaughan, 2013), and arboviruses (West Nile virus, La Crosse encephalitis virus; McJunkin et al, 1998;Turell et al, 2001;Haddow et al, 2011), any detected responses to logging in this area may influence the spatial distribution of mosquito-borne pathogens. We hypothesised that logging of intact temperate forests would alter the abundance and community composition of mosquitoes.…”