An Integrated Molecular Approach to Untangling Host-Vector-Pathogen Interactions in Mosquitoes (Diptera: Culicidae) From Sylvan Communities in Mexico.

There are ~240 species of Culicidae in Mexico, of which some are vectors of arthropod-borne viruses such as Zika virus, dengue virus, chikungunya virus, and West Nile virus. Thus, the identification of mosquito feeding preferences is paramount to understanding of vector-host-pathogen interactions that, in turn, can aid the control of disease outbreaks. Typically, DNA and RNA are extracted separately for animal (insects and blood meal hosts) and viral identification, but this study demonstrates that multiple organisms can be analyzed from a single RNA extract. For the first time, residual DNA present in standard RNA extracts was analyzed by DNA barcoding in concert with Sanger and next-generation sequencing (NGS) to identify both the mosquito species and the source of their meals in blood-fed females caught in seven sylvan communities in Chiapas State, Mexico. While mosquito molecular identification involved standard barcoding methods, the sensitivity of blood meal identification was maximized by employing short primers with NGS. In total, we collected 1,634 specimens belonging to 14 genera, 25 subgenera, and 61 morphospecies of mosquitoes. Of these, four species were new records for Mexico (Aedes guatemala, Ae. insolitus, Limatus asulleptus, Trichoprosopon pallidiventer), and nine were new records for Chiapas State. DNA barcode sequences for >300 bp of the COI gene were obtained from 291 specimens, whereas 130 bp sequences were recovered from another 179 specimens. High intraspecific divergence values (>2%) suggesting cryptic species complexes were observed in nine taxa: Anopheles eiseni (5.39%), An. pseudopunctipennis (2.79%), Ae. podographicus (4.05%), Culex eastor (4.88%), Cx. erraticus (2.28%), Toxorhynchites haemorrhoidalis (4.30%), Tr. pallidiventer (4.95%), Wyeomyia adelpha/Wy. guatemala (7.30%), and Wy. pseudopecten (4.04%). The study increased the number of mosquito species known from 128 species to 138 species for Chiapas State, and 239 for Mexico as a whole. Blood meal analysis showed that Aedes angustivittatus fed on ducks and chicken, whereas Psorophora albipes fed on humans. Culex quinquefasciatus fed on diverse hosts including chicken, human, turkey, and Mexican grackle. No arbovirus RNA was detected by reverse transcriptase-polymerase chain reaction in the surveyed specimens. This study demonstrated, for the first time, that residual DNA present in RNA blood meal extracts can be used to identify host vectors, highlighting the important role of molecular approaches in both vector identification and revealing host-vector-pathogen interactions.

The mosquitoes (Diptera: Culicidae) of Tabasco, Mexico.

To document and update the mosquito species of Tabasco, Mexico, field collection trips were conducted in the two physiographic regions of Tabasco: the coastal plain of the southern gulf and the mountains of Chiapas and Guatemala. Mosquitoes were collected as immature and adult stages during the dry and rainy seasons from 2014 through 2015. Additionally, the Reference Collection of Arthropods of Medical Importance (CAIM-InDRE) containing mosquitoes of Tabasco was re-examined. In total, 4,913 specimens were collected and examined, which are divided into seven tribes, 18 genera, 27 subgenera, and 104 species. Of these, one genus (Shannoniana Lane and Cerqueira), two subgenera (Georgecraigius Reinert, Harbach and Kitching, and Carrollia Lutz), and 21 species are new records for the mosquito fauna of Tabasco. Culex metempsytus Dyar is a new record for Mexico and Wyeomyia jocosa (Dyar and Knab) is removed from the Mexican mosquito fauna. Seventeen species historically reported were not found in the field collections conducted here. Taxonomic notes, new distribution limits, and comments about the medical importance of species of mosquitoes of Tabasco are discussed. Tabasco is the second state in Mexico with the largest mosquito richness (104 species), followed by Veracruz with 139 species.

First Record of Invasive Mosquito Aedes albopictus in Tabasco and Yucatan, Mexico.

The invasive mosquito Aedes albopictus is currently distributed in most of the southern Mexican region. Since the species was first recorded in the state of Tamaulipas, in northeastern Mexico in 1988, it has expanded its distribution throughout the Sierra Madre Oriental and Gulf of Mexico to the Neotropical region of the country. Currently the species occurs in the states of Tamaulipas, Coahuila, Nuevo Leon, Veracruz, Chiapas, Morelos, Quintana Roo, Sinaloa, San Luis Potosi, and Hidalgo. This is the first report of the mosquito in the states of Tabasco and Yucatan and the confirmation of its presence in Quintana Roo state. Aedes albopictus has been incriminated as a secondary vector of diseases such as those caused by dengue, chikungunya, and Zika viruses, which have caused epidemic outbreaks in most tropical and subtropical regions of Mexico; therefore, surveillance for the detection of Ae. albopictus is paramount so that targeted control strategies can be implemented for its control throughout Mexico.