A Follow-Up to the Geographical Distribution of Anopheles Species in Malaria-Endemic and Non-Endemic Areas of Honduras.

Anopheles species are the vectors of malaria, one of the diseases with the greatest impact on the health of the inhabitants of the tropics. Due to their epidemiological relevance and biological complexity, monitoring of anopheline populations in current and former malaria-endemic areas is critical for malaria risk assessment. Recent efforts have described the anopheline species present in the main malaria foci in Honduras. This study updates and expands knowledge about Anopheles species composition, geographical distribution, and genetic diversity in the continental territory of Honduras as in the Bay Islands. Outdoor insect collections were carried out at 25 sites in eight municipalities in five departments of Honduras between 2018 and 2021. Specimens were identified using taxonomic keys. Partial COI gene sequences were used for molecular species identification and phylogenetic analyses. In addition, detection of Plasmodium DNA was carried out in 255 female mosquitoes. Overall, 288 Anopheles mosquitoes were collected from 8 municipalities. Eight species were morphologically identified. Anopheles albimanus was the most abundant and widely distributed species (79.5%). A subset of 175 partial COI gene sequences from 8 species was obtained. Taxonomic identifications were confirmed via sequence analysis. Anopheles albimanus and An. apicimacula showed the highest haplotype diversity and nucleotide variation, respectively. Phylogenetic clustering was found for An. argyritarsis and An. neomaculipalpus when compared with mosquitoes from other Neotropical countries. Plasmodium DNA was not detected in any of the mosquitoes tested. This report builds upon recent records of the distribution and diversity of Anopheles species in malaria-endemic and non-endemic areas of Honduras. New COI sequences are reported for three anopheline species. This is also the first report of COI sequences of An. albimanus collected on the island of Roatán with apparent gene flow relative to mainland populations.

Distribution and phylogenetic diversity of Anopheles species in malaria endemic areas of Honduras in an elimination setting.

BACKGROUND: Anopheles mosquitoes are the vectors of malaria, one of the most important infectious diseases in the tropics. More than 500 Anopheles species have been described worldwide, and more than 30 are considered a public health problem. In Honduras, information on the distribution of Anopheles spp. and its genetic diversity is scarce. This study aimed to describe the distribution and genetic diversity of Anopheles mosquitoes in Honduras. METHODS: Mosquitoes were captured in 8 locations in 5 malaria endemic departments during 2019. Two collection methods were used. Adult anophelines were captured outdoors using CDC light traps and by aspiration of mosquitoes at rest. Morphological identification was performed using taxonomic keys. Genetic analyses included the sequencing of a partial region of the cytochrome c oxidase 1 gene (cox1) and the ribosomal internal transcribed spacer 2 (ITS2). RESULTS: A total of 1320 anophelines were collected and identified through morphological keys. Seven Anopheles species were identified. Anopheles albimanus was the most widespread and abundant species (74.02%). To confirm the morphological identification of the specimens, 175 and 122 sequences were obtained for cox1 and ITS2, respectively. Both markers confirmed the morphological identification. cox1 showed a greater nucleotide diversity than ITS2 in all species. High genetic diversity was observed within the populations of An. albimanus while An. darlingi proved to be a highly homogeneous population. Phylogenetic analyses revealed clustering patterns in An. darlingi and An. neivai in relation to specimens from South America. New sequences for An. crucians, An. vestitipennis and An. neivai are reported in this study. CONCLUSIONS: Here we report the distribution and genetic diversity of Anopheles species in endemic areas of malaria transmission in Honduras. According to our results, both taxonomic and molecular approaches are useful tools in the identification of anopheline mosquitoes. However, both molecular markers differ in their ability to detect intraspecific genetic diversity. These results provide supporting data for a better understanding of the distribution of malaria vectors in Honduras.

Blood Meal Sources of Anopheles spp. in Malaria Endemic Areas of Honduras.

Malaria remains a life-threatening disease in many tropical countries. Honduras has successfully reduced malaria transmission as different control methods have been applied, focusing mainly on indoor mosquitoes. The selective pressure exerted by the use of insecticides inside the households could modify the feeding behavior of the mosquitoes, forcing them to search for available animal hosts outside the houses. These animal hosts in the peridomicile could consequently become an important factor in maintaining vector populations in endemic areas. Herein, we investigated the blood meal sources and Plasmodium spp. infection on anophelines collected outdoors in endemic areas of Honduras. Individual PCR reactions with species-specific primers were used to detect five feeding sources on 181 visibly engorged mosquitoes. In addition, a subset of these mosquitoes was chosen for pathogen analysis by a nested PCR approach. Most mosquitoes fed on multiple hosts (2 to 4), and 24.9% of mosquitoes had fed on a single host, animal or human. Chicken and bovine were the most frequent blood meal sources (29.5% and 27.5%, respectively). The average human blood index (HBI) was 22.1%. None of the mosquitoes were found to be infected with Plasmodium spp. Our results show the opportunistic and zoophilic behavior of Anopheles mosquitoes in Honduras.

Absence of mutations associated with resistance to benzimidazole in the beta-tubulin gene of Ascaris suum.

INTRODUCTION: Benzimidazoles are commonly used for the control of veterinary nematodes. Resistance to benzimidazoles has been associated with three single nucleotide polymorphisms in the ß-tubulin gene of common nematodes. However, these mutations are infrequent in the genus Ascaris spp. METHODS: In order to determine mutations associated with benzimidazole resistance in Ascaris suum, worms were collected from slaughtered pigs and a partial region of the ß-tubulin gene was sequenced. RESULTS: All parasites showed the wildtype genotype for codons 167, 198, and 200 of the ß-tubulin gene. CONCLUSIONS: This is the first report of genetic sequences associated with benzimidazole resistance in A. suum.

Absence of mutations associated with resistance to benzimidazole in the beta-tubulin gene of Ascaris suum

Abstract INTRODUCTION: Benzimidazoles are commonly used for the control of veterinary nematodes. Resistance to benzimidazoles has been associated with three single nucleotide polymorphisms in the β-tubulin gene of common nematodes. However, these mutations are infrequent in the genus Ascaris spp. METHODS: In order to determine mutations associated with benzimidazole resistance in Ascaris suum, worms were collected from slaughtered pigs and a partial region of the β-tubulin gene was sequenced. RESULTS: All parasites showed the wildtype genotype for codons 167, 198, and 200 of the β-tubulin gene. CONCLUSIONS: This is the first report of genetic sequences associated with benzimidazole resistance in A. suum.