Entomological Survey and Leishmania (Leishmania) mexicana Prevalence in Sand Fly Species during an Outbreak of Cutaneous Leishmaniasis in Quintana Roo State, Mexico.

(1) Background: Localized cutaneous leishmaniasis is a neglected vector-borne disease that has become a serious public health problem in the Yucatan Peninsula. Although more than 60% of cases originate from the state of Quintana Roo, it is one of the least explored areas in terms of incriminating vectors of the Leishmania parasite. Additionally, cases of leishmaniasis have increased substantially in that region in recent years. For this reason, we explored and provided primary evidence of Leishmania DNA in sand fly species from four localities during outbreaks of leishmaniasis in Quintana Roo. We also contributed information on the regional genetic diversity of Leishmania parasites. (2) Methods: Sand flies were collected during several periods from November 2022 to April 2023 using Mosquito Light Circle and Shannon traps, as well as an active entomological search in refuges. For Leishmania detection, we amplified a fragment of 300-350 bp of the internal transcribed spacer subunit 1 (ITS-1). (3) Results: Of the 242 females collected, we detected Leishmania DNA in 25 specimens represented by Bichromomyia olmeca (1), Psathyromyia shannoni (17), Lutzomyia cruciata (4), Psathyromyia undulata (2), and Dampfomyia deleoni (1). The detection of Leishmania in these last two species represents new records for the Yucatan Peninsula and for Mexico. Leishmania (Leishmania) mexicana was the only species detected in the Phlebotominae species, with prevalence values that ranked between 7.41% and 33.33% from specimens collected in the sylvatic areas of Cozumel Island and Petcacab. (4) Conclusions: This study provides the first evidence of infection of Da. deleoni and Pa. undulata by L. (L.) Mexicana. In addition, the presence of three dominant haplotypes in all the evaluated localities was evidenced using the analysis of genetic diversity, and the locality of Petcacab was the one with the circulation of two new haplotypes not previously described in Mexico or neighboring countries. These results highlight the importance of intensive epidemiological surveillance due to the dynamics of transmission of Leishmania between different species.

Species diversity and detection of pathogens in phlebotomine sand flies collected from forest management areas of Quintana Roo, Mexico.

Sand flies have expanded their areas of distribution, thereby increasing the risk of pathogen transmission in non-endemic areas. To establish efficient prevention and control strategies for the transmission of vector-borne pathogens, it is important to understand seasonal dynamics of their vectors. In Mexico, there are several areas where the contact between sand flies, hosts and reservoirs favours the transmission of the pathogen. We compared sand fly communities in a forest management area and a conserved area in Noh-Bec, Quintana Roo, Mexico. The analysis included species diversity, activity peaks and molecular detection of pathogens. Sand flies were collected from November to December 2021 and April to May 2022, during 84 night-traps. The conserved area showed higher numbers and greater species heterogeneity of sand flies as compared with the other sites. The ß-diversity analysis revealed that sites disturbed by logging (S1, S2, S3) had greater similarity (90%) in their sand fly species composition than a conserved area (S4) (similarity = 36%). Although none of the specimens were infected with Leishmania, we detected Wolbachia (19.4%) in all four sites, as well as Bartonella (3.25%) only in the disturbed sites. Further studies on the dynamics of sand fly populations and their association with pathogens are necessary.

Phylogenetic relations among Mexican phlebotomine sand flies (Diptera: Psychodidae) and their divergence time estimation.

Phlebotomine sand flies (Diptera: Psychodidae: Phlebotominae) have biological relevance as vectors of several pathogens. To ensure periodic entomological monitoring it is necessary to have efficient and accurate tools for an adequate taxonomic identification. There are only few studies on phylogenetic analyses of phlebotomine sand flies from Neotropics, based mostly on morphological and/or molecular data, which makes the delimitation of intra- and interspecific variability of species challenging. Here we generated new molecular information on sand fly species distributed in endemic areas of leishmaniasis in Mexico, using mitochondrial and ribosomal genes, and incorporating morphological information available. Specifically, we established their phylogenetic relationships, and estimated their divergence time. Our study provides molecular information for 15 phlebotomine sand fly species from different areas of Mexico, contributing to the genetic inventory and phylogenetic relations among Neotropical species of the subfamily Phlebotominae. Mitochondrial genes proved to be suitable markers for the molecular identification of phlebotomine sand flies. However, the incorporation of additional nuclear gene information could increase the significance of phylogenetic inferences. We also provided evidence about a possible divergence time of phlebotomine sand fly species, supporting their presumable origin in the Cretaceous period.

New Distribution Records of Anopheles darlingi in Quintana Roo, Southeastern Mexico.

Anopheles darlingi is considered the main vector of malaria in the Neotropical region, so knowledge of its distribution in the Americas is highly relevant for the design of strategies for prevention and control of the illness. In Mexico, An. darlingi was recorded for the first time in 1943, and currently its distribution covers the states of Campeche, Chiapas, Quintana Roo, and Tabasco. In this study, new distribution data and observations of the abundance of An. darlingi in 14 localities of Quintana Roo, southeastern Mexico, are presented.

New Records and Updated Checklist of Mosquito Species in Quintana Roo, Mexico, Using DNA-Barcoding.

Collections of mosquitoes were conducted as part of the entomological vector surveillance in Quintana Roo State, Mexico, during September 2015. Species collected included Anopheles gabaldoni, An. darlingi, Psorophora columbiae, Culex inflictus, Cx. trifidus, Cx. lactator, and Wyeomyia guatemala s.l. All the specimens were identified by morphological and molecular characters (DNA-barcoding). This is the 1st time these species are reported in the Mexican state of Quintana Roo. This research updates and increases the list of species of mosquitoes in Quintana Roo from 79 to 86.

New Distribution Record of Aedes albopictus in Quintana Roo, Mexico, and its Importance to Public Health.

Aedes albopictus is an important vector of several arboviruses that affect human health worldwide; thus, knowledge of its distribution is a key factor for the implementation of disease control strategies. In Mexico, Ae. albopictus was recorded for the first time in 1988 in Tamaulipas State (northeast), but currently it has been recorded in 14 of the 32 states in the country. In 2012, it was recorded for the first time in a single locality in Quintana Roo (Cancún). In this study, we provide new distribution information for Ae. albopictus in the center and south of Quintana Roo State and comment on its medical importance.

Identification of phlebotomine sand flies (Diptera: Psychodidae) from leishmaniasis endemic areas in southeastern Mexico using DNA barcoding.

Leishmaniasis, a vector-borne disease transmitted to humans through the bite of phlebotomine sand flies, is of public health significance in southeastern Mexico. Active and continuous monitoring of vectors is an important aspect of disease control for the prediction of potential outbreaks. Thus, the correct identification of vectors is paramount in this regard. In this study, we employed DNA barcoding as a tool for identifying phlebotomine sand flies collected in localized cutaneous leishmaniasis endemic areas of Quintana Roo, Mexico. Specimens were collected using CDC light and Shannon traps as part of the Mexican Ministry of Health surveillance program. DNA extraction was carried out using a nondestructive protocol, and morphological identification based on taxonomic keys was conducted on slide-mounted specimens. Molecular taxonomic resolution using the 658-bp fragment of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene was 100% congruent with the morphological identification. Seven species were identified: Lutzomyia cruciata (Coquillett 1907), Lutzomyia longipalpis (Lutz & Neiva 1912), Psathyromyia shannoni (Dyar 1929), Dampfomyia deleoni (Fairchild & Hertig 1947), Dampfomyia beltrani/steatopyga (Vargas & Díaz-Nájera 1951), Bichromomyia olmeca olmeca (Vargas & Díaz-Nájera, 1959), and Brumptomyia mesai (Sherlock 1962). Mean intraspecific divergence ranged from 0.12% to 1.22%, while interspecific distances ranged from 11.59% to 19.29%. Neighbor-joining (NJ) analysis using the Kimura 2-parameter model also showed specimens of the same species to be clustered together. The study provides the first cox1 sequences for three species of sand flies and indicates the utility of DNA barcoding for phlebotomine sand flies species identification in southeastern Mexico.

DNA barcodes and evidence of cryptic diversity of anthropophagous mosquitoes in Quintana Roo, Mexico.

Culicidae mosquitoes are potential vectors of pathogens that affect human health. The correct species identification, as well as the discovery and description of cryptic species, is important in public health for the control and management of specific vectors. In the present study, the diversity of anthropophagous mosquitoes in Quintana Roo, at the border between Mexico and Belize, was evaluated using morphological and molecular data (COI-DNA Barcoding). A total of 1,413 adult female specimens were collected, belonging to eight genera and 31 morphospecies. Most species formed well-supported clades. Intraspecific Kimura 2 parameters (K2P) distance average was 0.75%, and a maximum distance of 4.40% was observed for Anopheles crucianss.l. ABGD method identified 28 entities, while 32 entities were identified with the BIN system. In Culex interrogator and Culex nigripalpus a low interspecific genetic distance of 0.1% was observed. One undescribed species belonging to the genus Aedes (Aedesn. sp.) was discovered, but no clear genetic divergence was found between this species and the closely related species Aedes angustivittatus. An intraspecific K2P distance greater than 2.7% was observed in Aedes serratus(3.9%), Anopheles crucianss.l. (4.4%), Culex taeniopus (3.7%), Haemagogus equinus (3.9%), Culex erraticus (5.0%), Psorophora ferox (4.5%), and in Anopheles apicimacula(8.10%); therefore, evidences of cryptic diversity are shown in these species. This study showed that DNA barcodes offer a reliable framework for mosquito species identification in Quintana Roo, except for some closely related species for which it is recommended to use additional nuclear genetic markers such as ITS2, in order to resolve these small discrepancies.

Confirmation of occurrence of Anopheles (Anopheles) veruslanei Vargas in Quintana Roo, Mexico using morphology and DNA barcodes.

In Mexico, genus Anopheles includes 27 species divided into three subgenera: Anopheles, Kerteszia, and Nyssorhynchus. Some species occur in the Nearctic region (northern Mexico), whereas other species occur in the Neotropical region (south and southeast Mexico) and only a few species occur in both regions. In Quintana Roo State (southeast Mexico) 11 species have been recorded: An. apicimacula, An. atropos, An. bradleyi, An. crucians, An. franciscanus, An. neomaculipalpus, An. pseudopunctipennis, An. punctimacula, An. veruslanei, An. vestitipennis and An. albimanus. However, the occurrence and identity of An. veruslanei has been questioned in recent years, since its description 39 years ago, it has not been reported in recent studies. In October 2015, five females of An. veruslanei were collected and identified. To corroborate their occurrence and identity in Quintana Roo State, we used morphological and molecular evidence that confirms it, and the type material of this species was studied to compare with the specimens of our collections.