erraticus, Cx. The overwintering mechanisms of these viruses are unknown. Edited chromatograms of host sequences had distinct and well-defined peaks with little background noise, indicating that the sequences were of good quality. The hole was positioned over the opening of the trap and a battery-powered aspirator, made from a modified 12V vacuum (BioQuip Products, Rancho Domiguez, CA), was turned on and held parallel to the hole, immediately outside the trap. Pens were set within the perimeter of a 60 × 30 × 4 m aviary-like enclosure (dark outer line). Alternatively, the artificial burrows inside the enclosures may provide suitable resting sites for mosquitoes that had fed from sleeping tegus, making them disinclined to leave the burrows before blood meals are digested. At this site, where P. bivittatus was an available host, Cx. Unlike northern Florida, the climate of southern Florida is also conducive to year-round mosquito activity, which would be expected to benefit the use of mosquito blood meals as a source of environmental DNA for P. bivittatus detection. 45 m), in Gainesville, Alachua County, Florida, USA. In Florida, its diet consists of both small and large animals including protected and managed species, such as wading birds, alligators and deer, and federally protected species [17–20]. Introduced mosquito species may affect vertebrate hosts and the transmission of local pathogens [68]. Data curation, here. We recovered python DNA from the blood meals of three species of Culex mosquitoes: Culex erraticus (Dyar and Knab), Culex quinquefasciatus Say, and Culex pilosus (Dyar and Knab). The intent of this study is to determine if local mosquito communities interact with P. bivittatus, an introduced host. We also collected blood-fed An. This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. erraticus, Cx. Investigation, The vegetation immediately surrounding the larger enclosure consisted of a pine flatwoods community with a canopy of Pinus palustris, and Pinus elliotti, interspersed with hardwoods, and an understory dominated by saw palmetto. Yes McGuire Center for Lepidoptera and Biodiversity, Florida Museum of Natural History, University of Florida, Gainesville, Florida, United States of America, Roles We observed other mosquito species (including Mansonia sp., Coquillettidia perturbans, Ps. A small sample of unfed female and male mosquitoes were collected to serve as negative controls to monitor for contamination and co-amplification of non-host templates. https://doi.org/10.1371/journal.pone.0190633.g002. Currently, at least 180 non-native herpetofaunal taxa have been introduced to the state and 63 are established, more than in any other U.S. state or global region [12]. A subset of the collected male and unfed female mosquitoes (n = 23, representing all species collected) were processed identically and served as negative controls, with the contents of the abdomen spread over the sampling area of the FTA card with a sterile pipette tip. Recent evidence suggests that snakes may be important to the transmission dynamics and overwintering of Eastern equine encephalitis virus [27, 28, 35, 60] and West Nile virus [61]. No, PLOS is a nonprofit 501(c)(3) corporation, #C2354500, based in San Francisco, California, US, https://doi.org/10.1371/journal.pone.0190633, Things that eat things you wouldn't expect them to. Edited sequences were searched on the National Center for Biotechnology Information (NCBI) GenBank database (https://www.ncbi.nlm.nih.gov/genbank/) using the Basic Local Alignment Search Tool (BLAST). Additional work, particularly field study, is needed to examine mosquito-python interactions and implications in nature in the Greater Everglades Ecosystem. To confirm that these blood meals were derived from A. carolinensis, their sequences were subsequently aligned to corresponding sequences obtained from A. carolinensis museum specimens (UF-Herpetology 170869, 170871), resulting in ≥99% homology. DNA was extracted following the HotSHOT method [42] using hot sodium hydroxide to lyse cells and a Tris-hydrogen chloride neutralization buffer solution. We speculate that python-mediated declines in or extirpations of mammal species could affect the transmission and prevalence of Everglades virus in southern Florida by increasing or decreasing contact between vector mosquitoes and reservoir hosts. Most research on its ecological impacts focuses on its role as a predator and its trophic interactions with native vertebrate species, particularly mammals. Native to Southeast Asia, pythons were first brought to the United States as exotic pets. Gut contents of blood fed mosquitoes were preserved on Whatman® four-sample Flinders Technology Associates (FTA) cards [40]. We thank Kandy L. Keacher and the staff of the USDA National Wildlife Research Center for assistance with collecting mosquito specimens, Lois Wood of the University of Florida Veterinary Entomology Lab, and Leroy Nuñez of the Florida Museum of Natural History Herpetology Division. crucians specimen sampled. While no known data are available on the pathogens of P. bivittatus in its native or introduced range, squamates (lizards and snakes) in Florida are the hosts of many mosquito-borne parasites including species of the taxonomically diverse genera Hepatozoon (Apicomplexa: Adeleorina) and Plasmodium (Apicomplexa: Haemosporida) [29–31]. In much of southern Florida, P. bivittatus is an available host for a diverse mosquito fauna (Fig 1) that includes more than 50 native and introduced mosquito species, some of which are potential pathogen vectors [26].
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