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Unbiased Characterization of Anopheles Mosquito Blood Meals by Targeted High-Throughput Sequencing

Cannon, Matthew V, Keven, John Bosco, Logue, Kyle, Reimer, Lisa ORCID:, Serre, David, Siba, Peter, Wlker, Edward D and Zimmerman, Peter A (2016) 'Unbiased Characterization of Anopheles Mosquito Blood Meals by Targeted High-Throughput Sequencing'. PLoS Neglected Tropical Diseases, Vol 10, Issue 3, e0004512.


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Understanding mosquito host choice is important for assessing vector competence or identifying disease reservoirs. Unfortunately, the availability of an unbiased method for comprehensively evaluating the composition of insect blood meals is very limited, as most current molecular assays only test for the presence of a few pre-selected species. These approaches also have limited ability to identify the presence of multiple mammalian hosts in a single blood meal. Here, we describe a novel high-throughput sequencing method that enables analysis of 96 mosquitoes simultaneously and provides a comprehensive and quantitative perspective on the composition of each blood meal. We validated in silico that universal primers targeting the mammalian mitochondrial 16S ribosomal RNA genes (16S rRNA) should amplify more than 95% of the mammalian 16S rRNA sequences present in the NCBI nucleotide database. We applied this method to 442 female Anopheles punctulatus s. l. mosquitoes collected in Papua New Guinea (PNG). While human (52.9%), dog (15.8%) and pig (29.2%) were the most common hosts identified in our study, we also detected DNA from mice, one marsupial species and two bat species. Our analyses also revealed that 16.3% of the mosquitoes fed on more than one host. Analysis of the human mitochondrial hypervariable region I in 102 human blood meals showed that 5 (4.9%) of the mosquitoes unambiguously fed on more than one person. Overall, analysis of PNG mosquitoes illustrates the potential of this approach to identify unsuspected hosts and characterize mixed blood meals, and shows how this approach can be adapted to evaluate inter-individual variations among human blood meals. Furthermore, this approach can be applied to any disease-transmitting arthropod and can be easily customized to investigate non-mammalian host sources.

Item Type: Article
Additional Information: Subjects: SM Open Access: SM Conceived and designed the experiments: EDW PAZ DS. Performed the experiments: KL JBK. Analyzed the data: KL MVC. Contributed reagents/materials/analysis tools: LR PS EDW PAZ DS. Wrote the paper: KL JBK EDW PAZ DS.
Subjects: QX Parasitology > QX 20 Research (General)
QX Parasitology > QX 4 General works
QX Parasitology > QX 45 Host-parasite relations
QX Parasitology > Insects. Other Parasites > QX 515 Anopheles
Faculty: Department: Biological Sciences > Vector Biology Department
Digital Object Identifer (DOI):
Depositing User: Stacy Murtagh
Date Deposited: 23 Mar 2016 14:24
Last Modified: 09 Sep 2019 08:40


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