DNA Barcoding Analysis of Trinidad Haemagogus Mosquitoes Reveals Evidence for Putative New Species.

Background: Haemagogus janthinomys is a primary sylvan vector of yellow fever virus and the emerging Mayaro virus. However, despite its medical importance, there is a dearth of data on the molecular taxonomy of this mosquito species. Methods: In this study, DNA barcoding analysis was performed on 64 adult female mosquitoes from Trinidad morphologically identified as Hg. janthinomys. The mitochondrial cytochrome c oxidase I (COI) gene and ribosomal DNA internal transcribed spacer 2 (ITS2) region of the mosquitoes were PCR amplified and sequenced, and molecular phylogenies inferred. Results: The BLASTN analysis showed that only 20% (n = 13/66) of COI sequences had high similarity (>99% identity) to Hg. janthinomys and the remaining sequences had low similarity (<90% identity) to reference GenBank sequences. Phylogenetic analysis of COI sequences revealed the presence of four strongly supported groups, with one distinct clade that did not align with any reference sequences. Corresponding ITS2 sequences for samples in this distinct COI group clustered into three clades. Conclusions: These molecular findings suggest the existence of a putative new Haemagogus mosquito species and underscore the need for further, more in-depth investigations into the taxonomy and classification of the Haemagogus genus.

Characterization of the virome associated with Haemagogus mosquitoes in Trinidad, West Indies.

Currently, there are increasing concerns about the possibility of a new epidemic due to emerging reports of Mayaro virus (MAYV) fever outbreaks in areas of South and Central America. Haemagogus mosquitoes, the primary sylvan vectors of MAYV are poorly characterized and a better understanding of the mosquito's viral transmission dynamics and interactions with MAYV and other microorganisms would be important in devising effective control strategies. In this study, a metatranscriptomic based approach was utilized to determine the prevalence of RNA viruses in field-caught mosquitoes morphologically identified as Haemagogus janthinomys from twelve (12) forest locations in Trinidad, West Indies. Known insect specific viruses including the Phasi Charoen-like and Humaiata-Tubiacanga virus dominated the virome of the mosquitoes throughout sampling locations while other viruses such as the avian leukosis virus, MAYV and several unclassified viruses had a narrower distribution. Additionally, assembled contigs from the Ecclesville location suggests the presence of a unique uncharacterized picorna-like virus. Mapping of RNA sequencing reads to reference mitochondrial sequences of potential feeding host animals showed hits against avian and rodent sequences, which putatively adds to the growing body of evidence of a potentially wide feeding host-range for the Haemagogus mosquito vector.

Changing patterns in the distribution of the Mayaro virus vector Haemagogus species in Trinidad, West Indies.

The Mayaro virus disease (MAYVD) is an emerging mosquito borne zoonosis that was first reported on the island of Trinidad in 1954. The viral agent for this disease is known to presently be endemic to Central and South America. The enzootic cycle of the Mayaro virus (MAYV) is not fully characterized, though primates are thought to be the main reservoir with Haemagogus species of mosquitoes as the primary vector. This virus has been responsible for several sporadic cases of infections and limited outbreaks, but it is postulated that the MAYVD will become a major epidemic in the future, following in the steps of the recent pandemics caused by Chikungunya and Zika viruses. Mitigating possible major outbreaks of MAYVD in the future would require effective strategies for vector control, for which knowledge on the ecology and distribution of the Haemagogus mosquitoes would be vitally important. In Trinidad, Haemagogus species have only been reported in the northwestern peninsula of the island based on studies up to 1995. However, no recent investigations have been completed to determine the status of this important vector on the island. The aim of this study was to investigate the current spatial distribution of Haemagogus species in the island of Trinidad, West Indies. Adult Haemagogus (Hag.) mosquitoes and larvae were surveyed during a twenty-month period using human bait trapping and ovitraps in major forested areas on the island. Mosquito species were identified using classical taxonomic keys. Haemagogus species were widespread and found in all forest types surveyed. Hag. janthinomys (85.7%) was the most widely distributed and dominant species on the island. Lower levels of Hag. leucocelaneus (7.3%), Hag. equinus (6.4%) and Hag. celeste (0.6%) were also collected. Overall, the proportion of mosquitoes collected in the wet season (June-December) was 3.5 times more than in the dry season (January-May). Mangroves, young secondary forests, semi-evergreen and evergreen forest types had relatively high mean abundance levels of Haemagogus species as compared to deciduous and montane forests. Proximity analysis suggests that population settlements within a 1 km buffer of the forest peripherals may be at risk for any emerging arboviral disease associated with these mosquito vectors. Haemagogus species showed a much wider distribution in Trinidad as compared to previous reports from up to 20 years ago and were prevalent in areas with no known presence of non-human primates. Since the MAYV has been previously implicated in causing infections in vertebrate hosts like rodents, birds and small mammals, the findings of this study suggest that there may be alternative hosts and reservoirs of this virus in the sylvatic cycle in Trinidad, other than primates. This has significant epidemiological implications for mosquito-borne viral infections in the region.