Haplotypes of Aedes aegypti and Aedes albopictus larvae detected in 18 districts of Costa Rica

Haplotypes of Aedes aegypti and Aedes albopictus larvae detected in 18 districts of Costa Rica | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Haplotypes of Aedes aegypti and Aedes albopictus larvae detected in 18 districts of Costa Rica Edwin Beltre-Nuñez, Sonia Flores-Nuñez, Silvia Arguello-Vargas, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7255371/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background Aedes aegypti and Aedes albopictus are key vectors of arboviruses such as dengue, chikungunya, and Zika. Monitoring the genetic diversity of these mosquito species is essential for understanding their adaptation, distribution, and implications for disease control. This study aimed to identify and compare the haplotypes of Aedes aegypti and Aedes albopictus larvae in 18 districts of Costa Rica. Methods A cross-sectional, descriptive study was conducted from August to November 2021. Larvae were collected from 26 locations across 18 districts at elevations below 900 m above sea level. Morphological identification was followed by DNA extraction and amplification of the cytochrome oxidase I (COI) mitochondrial gene by Polymerase Chain Reaction. Sequences were aligned and compared with GenBank references. Phylogenetic analysis was conducted using the unweighted pair-group method with arithmetic averages (UPGMA) clustering method. Results Ae. aegypti was detected in all districts, and Aedes albopictus in 11. Three haplotypes of Ae. aegypti (H1, H2, H4) and five of Ae. albopictus (H3, H6, H26, H37, H67) were identified. H1 ( Ae. aegypti ) and H67 ( Ae. albopictus ) were the most widespread. Several haplotypes not previously reported in Costa Rica were found. Amino acid polymorphisms were noted at key COI positions, with distinct clades observed in phylogenetic trees. Conclusions This study confirms the genetic diversity of Aedes populations in Costa Rica and identifies previously unreported haplotypes. Further seasonal and geographically broader studies are recommended to track vector population dynamics and inform vector control strategies. Mosquitos vectors phylogenetics polymorphisms clades Figures Figure 1 Figure 2 Figure 3 Background There are approximately 3,500 species of mosquitoes distributed on all continents except Antarctica. Some of these species are important since they transmit diseases [ 1 ]. Aedes aegypti and Aedes albopictus belong to the Culicidae family of the Diptera order being biological vectors of more than 26 viruses, including dengue, chikungunya, and Zika virus, impacting public health and posing an economic burden on hospitals, especially in tropical and subtropical regions [ 2 – 3 ]. Approximately two-fifths of the global population is at risk of contracting dengue fever, and an estimated 500,000 people are affected by hemorrhagic disease annually, with mortality rates exceeding 20% [ 4 – 5 ]. The chikungunya virus can also present as hemorrhagic disease and causes meningoencephalitis and myocarditis [ 6 ]. Zika virus causes severe joint pain, Guillain–Barré syndrome, and congenital anomalies such as microcephaly [ 7 ]. Traditional strategies for controlling vectors that transmit arboviruses include the application of insecticides and elimination of containers with standing water to prevent oviposition by mosquitoes [ 8 – 9 ]. In Costa Rica, pyrethroid insecticides (deltamethrin and cypermethrin) and organophosphates (temephos) are used in fumigation campaigns [ 10 ]. New control measures include the release of sterile insects and the replacement of the population with transgenic mosquitoes incapable of transmitting arboviruses [ 5 ]. Aedes control programs seek to reduce the density of the adult mosquito population below a critical threshold where epidemic transmission is unlikely to occur [ 5 ]. For this reason, it is essential to understand dispersal patterns, propagation in the environment, and the factors that facilitate the adaptation of mosquitoes to new environments. It is hypothesized that genetic changes in insects play a role in the adaptation of mosquitoes to new environments, making it important to study the genetic diversity of these mosquitoes [ 11 ]. Studies of genetic variation in Aedes species have been based on investigations of various regions of mitochondrial DNA [ 12 – 13 ], with the mitochondrial gene encoding cytochrome oxidase I (COI) being one of the most common genes studied to show linear evolution and a relatively fast rate of evolution [ 14 ]. The first genetic studies of Aedes determined little genetic variation and clustering of populations by continent or country [ 15 , 16 – 17 ], but recently, different haplotypes have been reported in populations of Aedes in the same continent or country [ 18 , 19 – 20 ]. Moreover, studies carried out in Italy have revealed different haplotypes of Ae. albopictus associated with tropical or temperate regions [ 21 ], whereas Cameroon populations of Ae. albopictus are related to specimens originating in tropical zones rather than temperate or subtropical zones [ 22 ]. In Senegal, domestic populations of Ae. aegypti tend to have less genetic diversity and variation than jungle populations do [ 23 ]; however, in Cambodia, little genetic diversity is found in populations of Ae. aegypti due to natural factors (habitat availability) and human factors (insecticide use) [ 24 ]. Studies within the country about the diversity of Ae. albopictus and Ae. aegypti is scarce. In the district of La Virgen de Sarapiquí, Futami et al. found seven haplotypes of Ae. albopictus (H17, H37, H67, H68, H69, H70, H71), and in the district of Siquirres Gloria-Soria et al. reported two haplotypes of Ae. aegypti (haplotypes A and G) [ 14 , 25 ]. This study aimed to investigate the genetic diversity of Aedes aegypti and Aedes albopictus in 18 districts of Costa Rica. Methods Type of study and sample collection A cross-sectional descriptive study was carried out. Larvae of Ae. aegypti and Ae. albopictus at all stages (L1-L4) were collected, at random, from the streets and sidewalks of 26 locations in 18 districts located less than 900 m above sea level between August and November 2021. The larvae were collected by emptying the contents of the breeding sites and filtering the water through a fine-mesh sieve. The larvae were then placed in a plastic jar with water and a lid. Larger or impractical breeding sites, the larvae were captured by passing the sieve over the surface of the container or using plastic Pasteur pipettes. The larvae were placed in containers with water and transported alive to the laboratory. Taxonomic identification of the larvae The taxonomic identification of the Aedes larvae was carried out using the morphological keys described by Rueda [ 26 ], visualizing the thorax and the eighth abdominal segment with a stereoscope. DNA extraction and Polymerase chain reaction (PCR) DNA of an individual larva of Ae. aegypti and of Ae. albopictus from each district was extracted using the QIAamp DNA Mini Kit (Qiagen, Germany) following the manufacturer's recommendations. The extracted DNA quality was measured with a NanoDrop One spectrophotometer (Thermo Fisher Scientific, USA) and subsequently stored at -20°C until analysis by molecular testing. The protocol described by Escobar et al. [ 27 ] was used to amplify a 710 bp region of the cytochrome oxidase I (COI) gene of Ae. aegypti . The initiators used were LCO1490 (5´-GGTCAACAAATCATAAAGATATTGG-3´) and HCO2198 (5´-TAAACTTCAGGGTGACCAAAAATCA-3´). First, a reaction mixture consisting of 25 µL of DreamTaqTM Master Mix (2x) (Thermo Fisher Scientific, USA), 17 µL of nuclease-free water (Thermo Fisher Scientific, USA), 2 µL of each primer (10 µM), and 4 µL of DNA was prepared. The cycling conditions were as follows: an initial cycle at 95°C for 10 min, 37 cycles at 94°C for 1 min, 48°C for 1 min, and 72°C for 1 min; and a final cycle at 72°C for 7 min. For Ae. albopictus , the protocol described by Zhong et al. [ 28 ] involved amplification of a 1390 bp fragment of the COI gene using two pairs of overlapping primers: albo1454F (5′ GGTCAACAAATCATAAAGATATTGG 3′), albo2160R (5′ TAAACTTCTGGATGACCAAAAAATCA 3′), and albo2027F (5′ CCCGTATTAGCCGGAGCTAT 3′). We used 25 µL of DreamTaq™ Master Mix (2x) (Thermo Fisher Scientific, USA), 18.5 µl of nuclease-free water (Thermo Fisher Scientific, USA), 2 µl of each primer (10 µM) and 2.5 µl of DNA. The thermal cycler conditions were as follows: initial denaturation at 94°C for 3 min; followed by 35 cycles at 94°C for 30 sec, 55°C for 30 sec, and 72°C for 1 min, and a final extension at 72°C for 6 min. The amplicons were subjected to 1% agarose gel electrophoresis with GelRed (Biotium, USA) and visualized on a transilluminator (Analytik Jena, Germany). Sequencing and haplotype analysis The amplified products were sent to Macrogen Company (Seoul, South Korea) for sequencing. The sequences obtained were edited, and the mutation sites and amino acid sequences were determined with the BioEdit 7.2.5 program [ 29 ]. In addition, the sequences were compared with those available in the GenBank database using the NCBI program BLAST (Basic Local Alignment Search Tool) to verify that they corresponded with known Aedes aegypti and Aedes albopictus sequences with a nucleotide similarity percentage greater than 99%. Also, the sequences were included in a multiple alignment performed by ClustalW [ 30 ]. The MEGA 10.1.8 program [ 31 ] was used to plot the unweighted pair group method with arithmetic averages (UPGMA) [ 32 ] tree based on Tajima-Nei's genetic distance [ 33 ]. The percentage of replicate trees on which the associated taxa clustered together in the bootstrap test (10000 replicates) is shown next to the branches [ 34 ]. Results Ae. aegypti was found in all districts, and Ae. albopictus in 11 of the 18 districts. Analysis of Ae. aegypti (one per district) revealed three genetic variants, the H1, H2, and H4 haplotypes (Table 1 ). The H1 haplotype was the most abundant and distributed, found in 14 districts, whereas the H2 and H4 haplotypes were found in one and three districts, respectively (Fig. 1 A; Table 1 ). On the other hand, the 11 larvae of Ae. albopictus included five genetic variants, namely, haplotypes H3, H6, H26, H37, and H67. The most abundant haplotype was H67, which was found in five districts, and H37, which was found in three districts. The remaining haplotypes were each found in only one district. Table 2 compares the haplotypes of Ae. aegypti and Ae. albopictus that were found in the different districts of Costa Rica in 2021, with reports of the same haplotypes detected in other countries. Table 1 Haplotypes of Ae. aegypti and Ae. albopictus found in 18 districts of Costa Rica in 2021 District Locality Haplotypes Ae. aegypti Haplotypes Ae. albopictus Geographic coordinate Length : Latitude Buenos Aires Buenos Aires H1 H37 -83,204331 9,94525923 Daniel Flores Concepción H1 H6 -83,683834 9,37194 San Isidro de El General San Isidro de El General H1 H3 -83,708781 9,374551 Chomes Punta Morales H1 H37 -84,945572 10,072762 Tarcoles Playa Azul H1 H37 -84,633717 9,775853 Puerto Viejo Barrio El Naranjal H1 H67 -83,986452 10,452357 La Virgen La Chilera H1 H26 -84,6108727 10,26541 El Cairo Cairo H1 H67 -83,532904 10,119598 Guacimo Afriquita H1 H67 -83,4010031 10,1290357 Guápiles iamante H1 H67 -83,774632 10,211391 Rita La Rita H1 H67 -83,776214 10,265982 Pital Pital H4 NO -84.275609 10.449463 Jaco Jaco H1 NO -84.6269457 9.6273826 La Isabel La Mora H2 NO -83.676245 9.915485 La Suiza La Suiza H1 NO -83.609574 9.849404 Pavones Eslabón H1 NO -83.641261 9.880263 Turrialba Carmen La Lira H4 NO -83.676715 9.913490 Siquirres San Rafael H4 NO -83.5082671 10.1047255 NO: No Ae. albopictus found in the district. Table 2 Comparison of the haplotypes of Ae. aegypti and Ae. albopictus found in different districts of Costa Rica in 2021, with reports of haplotypes detected in other countries. The GenBank access code, nucleotide identity, and percentage of identity of these haplotypes are shown. Ae. aegypti District Haplotype GenBank access code Country Nucleotide identity (bp/bp) Percentage of identity (%) Buenos Aires H1 ON100811 Honduras 494/494 100% Daniel Flores San Isidro de El General Chomes Tárcoles Jacó La Suiza Pavones Puerto Viejo La Virgen Cairo Guácimo Guápiles Rita MK028227-MK028228 El Salvador 483/483 100% MK028230-MK028232 MK028240-MK028242 MK028256 MK028258-MK028260 La Isabel H2 KX171389-KX171390 Panama 471/471 100% Pital H4 MF443395-MF443397 Canada 650/650 100% Siquirres MF043259 Turrialba KY022526 Germany, MN733746-MN733753 New Caledonia MN733743, MN733745 ON100786-ON100795 Honduras 494/494 100% ON100797-ON100810 ON100812-ON100813 ON100815-ON100816 MK028219-MK028220 El Salvador 483/483 100% MK028222-MK028226 MK028229, MK028231 MK028235-MK028239 MK028244-MK028245 MK028248, MK028250 MK028254, MK028261 MG241352 Costa Rica 575/575 100% KX171394 Panama 471/471 100% Ae. albopictus District Haplotype GenBank access code Country Nucleotide identity (bp/bp) Percentage of identity (%) San Isidro de El General H3 KC690898.1 Japan 1390/1390 China Taiwan 100% Italy USA Daniel Flores H6 KX886307.1 China 542/542 100% La Virgen H26 KC690921.1 Singapore 1390/1390 100% Buenos Aires Chomes Tárcoles H37* KC690932.1 Italy 100% USA Costa Rica Panama 1317/1317 Puerto Viejo Cairo Guácimo Guápiles Rita H67* AB907796.1 Costa Rica 100% 1390/1390 * Haplotypes previously reported in Costa Rica. Table 3 Polymorphic sites of the haplotypes of Ae. aegypti and Ae. albopictus from Costa Rica found in the sequences of the COI gene. Ae. aegypti Polymorphism position Haplotype 33 168 234 237 246 318 330 351 435 480 501 579 618 630 636 H1 T G T C G A A T G G C A T G C H2 T A T C G A G C A A C A C G C H3 C A C T A G G C G A T G T A T Ae. albopictus Polymorphism position Haplotype 40 94 467 954 1140 1404 H3 A G T A C G H6 T G C - - - H26 A G T A T A H37 A G T G T G H67 A A T A T G Table 4 Polymorphisms with amino acid changes in different haplotypes of Ae. aegypti and Ae. albopictus from Costa Rica according to the COI gene sequence. Ae. aegypti Change position in amino acids Haplotype 168 246 579 H1 Methionine Methionine Isoleucine H2 Isoleucine Methionine Isoleucine H3 Isoleucine Isoleucine Methionine Ae. albopictus Change position in amino acids Haplotype 40 94 - H3 Isoleucine Valine - H6 Phenylalanine Valine - H26 Isoleucine Valine - H37 Isoleucine Valine - H67 Isoleucine Isoleucine - The COI gene sequences of H1, H2, and H4 haplotypes of Ae. aegypti showed four, two, and nine polymorphic sites, respectively (Table 3 A), with the amino acid sequence affecting the polymorphic sites at positions 168, 246, and 579. At position 168, H1 encoded a methionine, whereas H2 and H4 encoded isoleucine. At position 246, H1 and H2 encoded methionine, and H4 isoleucine; finally, at position 579, H1 and H2 encoded isoleucine, and H4 methionine (Table 4 A). The H3, H26, H37 and H67 haplotypes of Ae. albopictus presented a polymorphic site in their sequences, whereas the H6 haplotype presented two polymorphic sites (Table 3 B). The amino acid sequence of the polymorphic sites was established at positions 40 and 94. At position 40, H6 encoded a phenylalanine, while the rest of the haplotypes encoded an isoleucine, and at position 94, H67 encoded an isoleucine, while the rest encoded valine (Table 4 B). The phylogenetic tree based on the COI gene established three well-defined clades for Ae. aegypti (Fig. 2 ) and five well-defined clades for Ae. albopictus , presenting a clade for each haplotype (Fig. 3 ). Discussion The number of haplotypes of Ae. aegypti (n = 8) determined using a total of 18 larvae collected from 18 locations in Costa Rica was similar to that reported in El Salvador (10 haplotypes in a total of 84 samples) and Panama (13 haplotypes in a total of 122 samples), and these larvae were collected from six and 30 locations, respectively [ 35 – 36 ]. Compared with these reports, numerous Ae. aegypti larvae with different haplotypes were detected, although only one larva of each species was analyzed in each district due to economic limitations. In Ae. albopictus , the number of haplotypes detected in a small number of larvae (5 haplotypes in 11 larvae) was similar to that reported in Brazil (10 haplotypes in 163 samples from 14 localities) [ 37 ] and in Colombia (33 haplotypes in 163 samples and 87 haplotypes in 145 samples from 14 and six locations, respectively) [ 2 , 38 ]. This could be due to the climatic and geographical similarities between Costa Rica and the countries of Central and South America; if more larvae are collected in different regions of Costa Rica, greater diversity may be found [ 2 ]. In this study, there are two haplotypes of Ae. aegypti (H1 and H2) and three haplotypes of Ae. albopictus (H3, H6, and H26) are reported for the first time in Costa Rica. Each of these haplotypes was detected in only one district, except for haplotype 1 of Ae. aegypti , which was widely distributed in a total of 14 districts [ 39 ]. This haplotype has also been reported in El Salvador, where it is the most widely distributed haplotype in the country [ 35 ]. Likewise, in Honduras, it is the most abundant and widespread haplotype [ 35 ]. Haplotype 2 of Ae. aegypti was found only in the La Isabel district in the southern central region of the country and had been previously reported in Panama [ 36 ]. Haplotype 3 of Ae. aegypti was found in three districts (Pital, Siquirres, and Turrialba) and has already been reported previously in the Siquirres district [ 25 ]. In contrast with haplotypes 1 and 2 of Ae. aegypti , which have only been reported to date in El Salvador, Honduras, and Panama, haplotype 4 has also been found in countries such as Canada, New Caledonia, and Germany, in addition to Central America. Haplotypes 3 and 6 of Ae. albopictus were found in the southern districts of the country and have been previously reported only in Japan, Italy, USA (H3), and China (H6). The third newly discovered haplotype (H26) of Ae. albopictus was found in the northern districts of the country and has been previously reported in Singapore. The other two haplotypes detected (H37 and H67) had previously been found in the district of La Virgen de Sarapiquí [ 14 ]. In the present study, H37 was found to be distributed in the Pacific and Atlantic areas of the country, whereas H67 was found to be more widely distributed, but only in the Atlantic area. Our findings agree with those of Futami et al. [ 14 ], who reported a high diversity of haplotypes of Ae. albopictus (7 haplotypes) in La Virgen de Sarapiquí. We found an additional haplotype in the same district, reporting for the first time the presence of H26 in La Virgen. In studies carried out in Medellín, Colombia, different haplotypes were determined in three neighborhoods and two time periods (January–April 2010 and October–December 2012). The results revealed that the genetic composition changes occurred without a predictable pattern in 2 years [ 2 ]. Conclusion The genetic diversity of Aedes populations in Costa Rica was confirmed, and previously unreported haplotypes were reported. Seasonal and geographically broader studies are recommended to understand vector population dynamics. For example, conducting systematic studies with multiple larvae from each district in the dry and rainy seasons, to establish seasonal differences, which could increase our understanding of genetic diversity, reproduction speed, and genetic flow of natural mosquito populations. These analyses would provide not only an evolutionary perspective on vector populations but also a key tool for designing and optimizing control strategies that can slow the spread of diseases transmitted by arbovirus vectors. Declarations Acknowledgments The Costa Rican Ministry of Health, especially Rodrigo Marín, Manuel Gutiérrez, Gabriela Delgado, Carlos Aguilar, and the regional technicians who assisted with sample collection. The German Academic Exchange Service (DAAD) awarded scholarships to Sonia Flores Núñez and Edwin Beltré Nuñez. Funding The project was funded by FUNDAUNA “Diagnosis and Research in Population and Ecosystem Medicine” and Vicerrectoría de Investigación, Universidad Nacional. Availability of data and materials The data sets used during the current study are available from the corresponding authors on reasonable request. Authors contributions Each author's contribution to the manuscript is as follows: Edwin Beltré-Nuñez: conceptualization, methodology, formal analysis, investigation, data curation, writing - original draft. Sonia Flores-Nuñez: conceptualization, methodology, formal analysis, investigation, data curation, writing - original draft. Silvia Argüello-Vargas: conceptualization, methodology, investigation, resources, writing - original draft, writing - review, supervision, project administration, funding. Antony Solorzano-Morales: methodology, investigation, writing - review, editing. Gaby Dolz: conceptualization, methodology, investigation, resources, writing - original draft, writing - review, supervision, project administration, funding. Ethics approval and consent to participate, consent for publication, competing interests The authors declare that they have fully complied with all relevant ethical and legal requirements, both during the study and in the production of the manuscript; that there are no conflicts of interest of any kind; that all funding sources are fully and clearly stated in the acknowledgments section; and that they fully agree with the final edited version of the article. The authors declare no relevant or material financial interests that relate to the research described in this paper. References Aguirre-Obando AO, Navarro-Silva MA. How much is known about the genetic diversity of the Asian tiger mosquito? a systematic review. Rev Univ Ind Satander Salud. 2017; 49(3):422–437. http://dx.doi.org/10.18273/revsal.v49n3-2017001 Cadavid JM, Rúa G, Campo O, Bedoya G, Rojas W. Cambios genéticos temporales y microgeográficos de Aedes aegypti en Medellín, Colombia. 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Escobar D, Ortiz B, Urrutia O, Fontecha G. Genetic diversity among four populations of Aedes aegypti (Diptera: Culicidae ) from Honduras as revealed by mitochondrial DNA cytochrome oxidase I. Pathogens. 2022; 11(6): 620. doi: https://doi.org/10.3390/pathogens11060620 Zhong D, Lo E, Hu R, Metzger ME, Cummings R, Bonizzoni M, et al. Genetic analysis of invasive Aedes albopictus populations in Los Angeles County, California and its potential public health impact. PLoS One. 2013;8(7):e68586. doi: 10.1371/journal.pone.0068586 . PMID: 23861921; PMCID: PMC3702605. Hall TA. Bioedit: a user-friendly biological sequence alignment editor and analysis program for windows 95/98/NT. Nucleic Acids Symp Ser. 1999;41: 95–98. Thompson JD, Higgins DG, Gibson TJ. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 1994;22(22):4673–80. doi: 10.1093/nar/22.22.4673 . PMID: 7984417; PMCID: PMC308517. Kumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: molecular evolutionary genetics analysis across Computing Platforms. Mol Biol Evol. 2018;35(6):1547–1549. doi: 10.1093/molbev/msy096 . PMID: 29722887; PMCID: PMC5967553. Sneath PH, Sokal RR. Numerical taxonomy: the principles and practice of numerical classification. 1st ed. San Francisco: W. H. Freeman; 1973. Tajima F, Nei M. Estimation of evolutionary distance between nucleotide sequences. Mol Biol Evol. 1984;1(3):269 – 85. doi: 10.1093/oxfordjournals.molbev.a040317 . PMID: 6599968. Felsenstein J. Confidence limits on phylogenies: an approach using the bootstrap. Evolution. 1985;39(4):783–791. doi: 10.1111/j.1558-5646.1985.tb00420.x . PMID: 28561359. Joyce AL, Torres MM, Torres R, Moreno M. Genetic variability of the Aedes aegypti (Diptera: Culicidae ) mosquito in El Salvador, vector of dengue, yellow fever, chikungunya and Zika. Parasites Vectors. 2018;11(1):1–14. doi: https://doi.org/10.1186/s13071-018-3226-5 Eskildsen GA, Rovira JR, Smith O, Miller MJ, Bennett KL, McMillan WO. Maternal invasion history of Aedes aegypti and Aedes albopictus into the Isthmus of Panama: implications for the control of emergent viral disease agents. PLoS One. 2018;13(3). doi: https://doi.org/10.1371/journal.pone.0194874 Scarpassa VM, Cardoza TB, Cardoso RP. Population genetics and phylogeography of Aedes aegypti (Diptera: Culicidae ) from Brazil. ASTM. 2008;78(6):895–903. doi: https://doi.org/10.4269/ajtmh.2008.78.895 Gómez M, Martínez D, Hernández C, Luna N, Patiño LH, Melo RB, et al. Arbovirus infection in Aedes aegypti from different departments of Colombia. Front Ecol Evol. 2022; doi: 10.3389/fevo.2022.999169 . Barrientos O, Chaves G. Región Huetar Norte: oferta exportada actual y oferta potencial de productos agropecuarios alternativos (No. E71/10543). Promotora del Comercio Exterior de Costa Rica. 2008. http://www.mag.go.cr/bibliotecavirtual/E71-10543.pdf . Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7255371","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":503305584,"identity":"c62b723e-2791-41ff-9a75-56d09082d80b","order_by":0,"name":"Edwin Beltre-Nuñez","email":"","orcid":"","institution":"National University of Costa Rica","correspondingAuthor":false,"prefix":"","firstName":"Edwin","middleName":"","lastName":"Beltre-Nuñez","suffix":""},{"id":503305585,"identity":"ebe8b200-1715-4e82-8653-98e9d823a74e","order_by":1,"name":"Sonia Flores-Nuñez","email":"","orcid":"","institution":"National University of Costa Rica","correspondingAuthor":false,"prefix":"","firstName":"Sonia","middleName":"","lastName":"Flores-Nuñez","suffix":""},{"id":503305586,"identity":"d88b0248-b066-4285-8d11-422cc8c5d381","order_by":2,"name":"Silvia Arguello-Vargas","email":"","orcid":"","institution":"National University of Costa Rica","correspondingAuthor":false,"prefix":"","firstName":"Silvia","middleName":"","lastName":"Arguello-Vargas","suffix":""},{"id":503305587,"identity":"3c7ea734-7d2d-41e1-8e5e-153ed818eb7a","order_by":3,"name":"Antony Solorzano-Morales","email":"","orcid":"","institution":"National University of Costa Rica","correspondingAuthor":false,"prefix":"","firstName":"Antony","middleName":"","lastName":"Solorzano-Morales","suffix":""},{"id":503305588,"identity":"885dc0b4-6c9b-41da-89f7-ed83d8ada039","order_by":4,"name":"Gaby Dolz","email":"data:image/png;base64,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","orcid":"","institution":"National University of Costa Rica","correspondingAuthor":true,"prefix":"","firstName":"Gaby","middleName":"","lastName":"Dolz","suffix":""}],"badges":[],"createdAt":"2025-07-30 17:38:17","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7255371/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7255371/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":90041510,"identity":"4c769bb4-cf35-4653-88e0-5b1457e5885c","added_by":"auto","created_at":"2025-08-27 17:06:57","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":262422,"visible":true,"origin":"","legend":"\u003cp\u003eDistribution map of the haplotypes of \u003cem\u003eAe. aegypti\u003c/em\u003e(1A) and \u003cem\u003eAe. albopictus\u003c/em\u003e (1B) found in 18 and 11 districts of Costa Rica, respectively, in 2021.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-7255371/v1/58482e55edda48ca45403368.png"},{"id":90041509,"identity":"0ce63658-02ff-4491-858b-a3ecded52d4b","added_by":"auto","created_at":"2025-08-27 17:06:57","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":103932,"visible":true,"origin":"","legend":"\u003cp\u003eDendrogram obtained from a 650-nucleotide fragment of the COI gene of \u003cem\u003eAe. aegypti\u003c/em\u003e constructed using the UPGMA method. Three reference strains of \u003cem\u003eAe. aegypti\u003c/em\u003e and the haplotypes found in the present study are shown.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-7255371/v1/f231cf4476750381fd8f85eb.png"},{"id":90041512,"identity":"a04c457c-c6f9-4f21-afe5-0d082fcbf184","added_by":"auto","created_at":"2025-08-27 17:06:57","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":119325,"visible":true,"origin":"","legend":"\u003cp\u003eDendrogram obtained from a 1007-nucleotide fragment of the COI gene of \u003cem\u003eAe. albopictus\u003c/em\u003econstructed using the UPGMA method. Five reference strains of \u003cem\u003eAe. albopictus\u003c/em\u003e and the haplotypes found in the present study are shown.\u003c/p\u003e","description":"","filename":"floatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-7255371/v1/d7db0bb502324f7adc51fc1e.png"},{"id":95233106,"identity":"b0f4e21c-8c7a-4284-9987-eecda07fd9dc","added_by":"auto","created_at":"2025-11-05 16:41:46","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1456733,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7255371/v1/67a388b6-1f9d-492b-b2fd-950b6ea954c2.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Haplotypes of Aedes aegypti and Aedes albopictus larvae detected in 18 districts of Costa Rica","fulltext":[{"header":"Background","content":"\u003cp\u003eThere are approximately 3,500 species of mosquitoes distributed on all continents except Antarctica. Some of these species are important since they transmit diseases [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. \u003cem\u003eAedes aegypti\u003c/em\u003e and \u003cem\u003eAedes albopictus\u003c/em\u003e belong to the Culicidae family of the Diptera order being biological vectors of more than 26 viruses, including dengue, chikungunya, and Zika virus, impacting public health and posing an economic burden on hospitals, especially in tropical and subtropical regions [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e–\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Approximately two-fifths of the global population is at risk of contracting dengue fever, and an estimated 500,000 people are affected by hemorrhagic disease annually, with mortality rates exceeding 20% [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e–\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. The chikungunya virus can also present as hemorrhagic disease and causes meningoencephalitis and myocarditis [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Zika virus causes severe joint pain, Guillain–Barré syndrome, and congenital anomalies such as microcephaly [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eTraditional strategies for controlling vectors that transmit arboviruses include the application of insecticides and elimination of containers with standing water to prevent oviposition by mosquitoes [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e–\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. In Costa Rica, pyrethroid insecticides (deltamethrin and cypermethrin) and organophosphates (temephos) are used in fumigation campaigns [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. New control measures include the release of sterile insects and the replacement of the population with transgenic mosquitoes incapable of transmitting arboviruses [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003cem\u003eAedes\u003c/em\u003e control programs seek to reduce the density of the adult mosquito population below a critical threshold where epidemic transmission is unlikely to occur [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. For this reason, it is essential to understand dispersal patterns, propagation in the environment, and the factors that facilitate the adaptation of mosquitoes to new environments. It is hypothesized that genetic changes in insects play a role in the adaptation of mosquitoes to new environments, making it important to study the genetic diversity of these mosquitoes [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eStudies of genetic variation in \u003cem\u003eAedes\u003c/em\u003e species have been based on investigations of various regions of mitochondrial DNA [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e–\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e], with the mitochondrial gene encoding cytochrome oxidase I (COI) being one of the most common genes studied to show linear evolution and a relatively fast rate of evolution [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThe first genetic studies of \u003cem\u003eAedes\u003c/em\u003e determined little genetic variation and clustering of populations by continent or country [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e–\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e], but recently, different haplotypes have been reported in populations of \u003cem\u003eAedes\u003c/em\u003e in the same continent or country [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e–\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eMoreover, studies carried out in Italy have revealed different haplotypes of \u003cem\u003eAe. albopictus\u003c/em\u003e associated with tropical or temperate regions [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e], whereas Cameroon populations of \u003cem\u003eAe. albopictus\u003c/em\u003e are related to specimens originating in tropical zones rather than temperate or subtropical zones [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. In Senegal, domestic populations of \u003cem\u003eAe. aegypti\u003c/em\u003e tend to have less genetic diversity and variation than jungle populations do [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]; however, in Cambodia, little genetic diversity is found in populations of \u003cem\u003eAe. aegypti\u003c/em\u003e due to natural factors (habitat availability) and human factors (insecticide use) [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eStudies within the country about the diversity of \u003cem\u003eAe. albopictus\u003c/em\u003e and \u003cem\u003eAe. aegypti\u003c/em\u003e is scarce. In the district of La Virgen de Sarapiquí, Futami et al. found seven haplotypes of \u003cem\u003eAe. albopictus\u003c/em\u003e (H17, H37, H67, H68, H69, H70, H71), and in the district of Siquirres Gloria-Soria et al. reported two haplotypes of \u003cem\u003eAe. aegypti\u003c/em\u003e (haplotypes A and G) [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. This study aimed to investigate the genetic diversity of \u003cem\u003eAedes aegypti\u003c/em\u003e and \u003cem\u003eAedes albopictus\u003c/em\u003e in 18 districts of Costa Rica.\u003c/p\u003e"},{"header":"Methods","content":"\u003cp\u003e\u003cb\u003eType of study and sample collection\u003c/b\u003e\u003c/p\u003e\u003cp\u003eA cross-sectional descriptive study was carried out. Larvae of \u003cem\u003eAe. aegypti\u003c/em\u003e and \u003cem\u003eAe. albopictus\u003c/em\u003e at all stages (L1-L4) were collected, at random, from the streets and sidewalks of 26 locations in 18 districts located less than 900 m above sea level between August and November 2021. The larvae were collected by emptying the contents of the breeding sites and filtering the water through a fine-mesh sieve. The larvae were then placed in a plastic jar with water and a lid. Larger or impractical breeding sites, the larvae were captured by passing the sieve over the surface of the container or using plastic Pasteur pipettes. The larvae were placed in containers with water and transported alive to the laboratory.\u003c/p\u003e\u003cp\u003e\u003cb\u003eTaxonomic identification of the larvae\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe taxonomic identification of the \u003cem\u003eAedes\u003c/em\u003e larvae was carried out using the morphological keys described by Rueda [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e], visualizing the thorax and the eighth abdominal segment with a stereoscope.\u003c/p\u003e\u003cp\u003e\u003cb\u003eDNA extraction and Polymerase chain reaction (PCR)\u003c/b\u003e\u003c/p\u003e\u003cp\u003eDNA of an individual larva of \u003cem\u003eAe. aegypti\u003c/em\u003e and of \u003cem\u003eAe. albopictus\u003c/em\u003e from each district was extracted using the QIAamp DNA Mini Kit (Qiagen, Germany) following the manufacturer's recommendations. The extracted DNA quality was measured with a NanoDrop One spectrophotometer (Thermo Fisher Scientific, USA) and subsequently stored at -20°C until analysis by molecular testing.\u003c/p\u003e\u003cp\u003eThe protocol described by Escobar et al. [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e] was used to amplify a 710 bp region of the cytochrome oxidase I (COI) gene of \u003cem\u003eAe. aegypti\u003c/em\u003e. The initiators used were LCO1490 (5´-GGTCAACAAATCATAAAGATATTGG-3´) and HCO2198 (5´-TAAACTTCAGGGTGACCAAAAATCA-3´). First, a reaction mixture consisting of 25 µL of DreamTaqTM Master Mix (2x) (Thermo Fisher Scientific, USA), 17 µL of nuclease-free water (Thermo Fisher Scientific, USA), 2 µL of each primer (10 µM), and 4 µL of DNA was prepared. The cycling conditions were as follows: an initial cycle at 95°C for 10 min, 37 cycles at 94°C for 1 min, 48°C for 1 min, and 72°C for 1 min; and a final cycle at 72°C for 7 min.\u003c/p\u003e\u003cp\u003eFor \u003cem\u003eAe. albopictus\u003c/em\u003e, the protocol described by Zhong et al. [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e] involved amplification of a 1390 bp fragment of the COI gene using two pairs of overlapping primers: albo1454F (5′ GGTCAACAAATCATAAAGATATTGG 3′), albo2160R (5′ TAAACTTCTGGATGACCAAAAAATCA 3′), and albo2027F (5′ CCCGTATTAGCCGGAGCTAT 3′). We used 25 µL of DreamTaq™ Master Mix (2x) (Thermo Fisher Scientific, USA), 18.5 µl of nuclease-free water (Thermo Fisher Scientific, USA), 2 µl of each primer (10 µM) and 2.5 µl of DNA. The thermal cycler conditions were as follows: initial denaturation at 94°C for 3 min; followed by 35 cycles at 94°C for 30 sec, 55°C for 30 sec, and 72°C for 1 min, and a final extension at 72°C for 6 min.\u003c/p\u003e\u003cp\u003eThe amplicons were subjected to 1% agarose gel electrophoresis with GelRed (Biotium, USA) and visualized on a transilluminator (Analytik Jena, Germany).\u003c/p\u003e\u003cp\u003e\u003cb\u003eSequencing and haplotype analysis\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe amplified products were sent to Macrogen Company (Seoul, South Korea) for sequencing. The sequences obtained were edited, and the mutation sites and amino acid sequences were determined with the BioEdit 7.2.5 program [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. In addition, the sequences were compared with those available in the GenBank database using the NCBI program BLAST (Basic Local Alignment Search Tool) to verify that they corresponded with known \u003cem\u003eAedes aegypti\u003c/em\u003e and \u003cem\u003eAedes albopictus\u003c/em\u003e sequences with a nucleotide similarity percentage greater than 99%. Also, the sequences were included in a multiple alignment performed by ClustalW [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. The MEGA 10.1.8 program [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e] was used to plot the unweighted pair group method with arithmetic averages (UPGMA) [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e] tree based on Tajima-Nei's genetic distance [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. The percentage of replicate trees on which the associated taxa clustered together in the bootstrap test (10000 replicates) is shown next to the branches [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e].\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cem\u003eAe. aegypti\u003c/em\u003e was found in all districts, and \u003cem\u003eAe. albopictus\u003c/em\u003e in 11 of the 18 districts. Analysis of \u003cem\u003eAe. aegypti\u003c/em\u003e (one per district) revealed three genetic variants, the H1, H2, and H4 haplotypes (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The H1 haplotype was the most abundant and distributed, found in 14 districts, whereas the H2 and H4 haplotypes were found in one and three districts, respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA; Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). On the other hand, the 11 larvae of \u003cem\u003eAe. albopictus\u003c/em\u003e included five genetic variants, namely, haplotypes H3, H6, H26, H37, and H67. The most abundant haplotype was H67, which was found in five districts, and H37, which was found in three districts. The remaining haplotypes were each found in only one district. Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e compares the haplotypes of \u003cem\u003eAe. aegypti\u003c/em\u003e and \u003cem\u003eAe. albopictus\u003c/em\u003e that were found in the different districts of Costa Rica in 2021, with reports of the same haplotypes detected in other countries.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eHaplotypes of \u003cem\u003eAe. aegypti\u003c/em\u003e and \u003cem\u003eAe. albopictus\u003c/em\u003e found in 18 districts of Costa Rica in 2021\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"6\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eDistrict\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eLocality\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eHaplotypes \u003cem\u003eAe. aegypti\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eHaplotypes\u003c/p\u003e\u003cp\u003e\u003cem\u003eAe. albopictus\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e\u003cp\u003eGeographic coordinate\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003eLength\u003c/b\u003e:\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003eLatitude\u003c/b\u003e\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBuenos Aires\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eBuenos Aires\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eH1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eH37\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-83,204331\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e9,94525923\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDaniel Flores\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eConcepci\u0026oacute;n\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eH1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eH6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-83,683834\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e9,37194\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSan Isidro de El General\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSan Isidro de El General\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eH1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eH3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-83,708781\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e9,374551\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eChomes\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePunta Morales\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eH1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eH37\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-84,945572\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e10,072762\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTarcoles\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePlaya Azul\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eH1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eH37\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-84,633717\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e9,775853\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePuerto Viejo\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eBarrio El Naranjal\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eH1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eH67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-83,986452\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e10,452357\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLa Virgen\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLa Chilera\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eH1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eH26\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-84,6108727\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e10,26541\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eEl Cairo\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCairo\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eH1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eH67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-83,532904\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e10,119598\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGuacimo\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAfriquita\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eH1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eH67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-83,4010031\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e10,1290357\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGu\u0026aacute;piles\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eiamante\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eH1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eH67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-83,774632\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e10,211391\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRita\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLa Rita\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eH1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eH67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-83,776214\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e10,265982\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePital\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePital\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eH4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eNO\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-84.275609\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e10.449463\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eJaco\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eJaco\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eH1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eNO\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-84.6269457\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e9.6273826\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLa Isabel\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLa Mora\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eH2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eNO\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-83.676245\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e9.915485\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLa Suiza\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLa Suiza\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eH1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eNO\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-83.609574\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e9.849404\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePavones\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eEslab\u0026oacute;n\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eH1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eNO\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-83.641261\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e9.880263\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTurrialba\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCarmen La Lira\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eH4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eNO\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-83.676715\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e9.913490\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSiquirres\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSan Rafael\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eH4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eNO\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-83.5082671\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e10.1047255\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"6\"\u003eNO: No \u003cem\u003eAe. albopictus\u003c/em\u003e found in the district.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eComparison of the haplotypes of \u003cem\u003eAe. aegypti\u003c/em\u003e and \u003cem\u003eAe. albopictus\u003c/em\u003e found in different districts of Costa Rica in 2021, with reports of haplotypes detected in other countries. The GenBank access code, nucleotide identity, and percentage of identity of these haplotypes are shown.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"6\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colspan=\"6\" nameend=\"c6\" namest=\"c1\"\u003e\u003cp\u003e\u003cem\u003eAe. aegypti\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eDistrict\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eHaplotype\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003eGenBank access code\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003eCountry\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003eNucleotide identity (bp/bp)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003ePercentage of identity (%)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBuenos Aires\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\" morerows=\"18\" rowspan=\"19\"\u003e\u003cp\u003eH1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\" morerows=\"13\" rowspan=\"14\"\u003e\u003cp\u003eON100811\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\" morerows=\"13\" rowspan=\"14\"\u003e\u003cp\u003eHonduras\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\" morerows=\"13\" rowspan=\"14\"\u003e\u003cp\u003e494/494\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\" morerows=\"13\" rowspan=\"14\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDaniel Flores\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSan Isidro de El General\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eChomes\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eT\u0026aacute;rcoles\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eJac\u0026oacute;\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLa Suiza\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePavones\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePuerto Viejo\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLa Virgen\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCairo\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGu\u0026aacute;cimo\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGu\u0026aacute;piles\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRita\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMK028227-MK028228\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\" morerows=\"4\" rowspan=\"5\"\u003e\u003cp\u003eEl Salvador\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\" morerows=\"4\" rowspan=\"5\"\u003e\u003cp\u003e483/483\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\" morerows=\"4\" rowspan=\"5\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMK028230-MK028232\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMK028240-MK028242\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMK028256\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMK028258-MK028260\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLa Isabel\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eH2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eKX171389-KX171390\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ePanama\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e471/471\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePital\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\" morerows=\"17\" rowspan=\"18\"\u003e\u003cp\u003eH4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMF443395-MF443397\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eCanada\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\" morerows=\"4\" rowspan=\"5\"\u003e\u003cp\u003e650/650\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\" morerows=\"4\" rowspan=\"5\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSiquirres\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMF043259\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTurrialba\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eKY022526\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eGermany,\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMN733746-MN733753\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eNew Caledonia\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMN733743, MN733745\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eON100786-ON100795\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\" morerows=\"3\" rowspan=\"4\"\u003e\u003cp\u003eHonduras\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\" morerows=\"3\" rowspan=\"4\"\u003e\u003cp\u003e494/494\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\" morerows=\"3\" rowspan=\"4\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eON100797-ON100810\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eON100812-ON100813\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eON100815-ON100816\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMK028219-MK028220\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\" morerows=\"6\" rowspan=\"7\"\u003e\u003cp\u003eEl Salvador\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\" morerows=\"6\" rowspan=\"7\"\u003e\u003cp\u003e483/483\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\" morerows=\"6\" rowspan=\"7\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMK028222-MK028226\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMK028229, MK028231\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMK028235-MK028239\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMK028244-MK028245\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMK028248, MK028250\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMK028254, MK028261\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMG241352\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eCosta Rica\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e575/575\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eKX171394\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ePanama\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e471/471\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"6\" nameend=\"c6\" namest=\"c1\"\u003e\u003cp\u003e\u003cb\u003eAe. albopictus\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eDistrict\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eHaplotype\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003eGenBank access code\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003eCountry\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003eNucleotide identity (bp/bp)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003ePercentage of identity (%)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e\u003cp\u003eSan Isidro de El General\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\" morerows=\"4\" rowspan=\"5\"\u003e\u003cp\u003eH3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\" morerows=\"4\" rowspan=\"5\"\u003e\u003cp\u003eKC690898.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eJapan\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\" morerows=\"4\" rowspan=\"5\"\u003e\u003cp\u003e1390/1390\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eChina\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eTaiwan\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eItaly\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eUSA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDaniel Flores\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eH6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eKX886307.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eChina\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e542/542\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLa Virgen\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eH26\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eKC690921.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eSingapore\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e1390/1390\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e\u003cp\u003eBuenos Aires\u003c/p\u003e\u003cp\u003eChomes\u003c/p\u003e\u003cp\u003eT\u0026aacute;rcoles\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\" morerows=\"2\" rowspan=\"3\"\u003e\u003cp\u003eH37*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\" morerows=\"2\" rowspan=\"3\"\u003e\u003cp\u003eKC690932.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eItaly\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\" morerows=\"2\" rowspan=\"3\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eUSA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eCosta Rica Panama\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e1317/1317\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003ePuerto Viejo\u003c/p\u003e\u003cp\u003eCairo\u003c/p\u003e\u003cp\u003eGu\u0026aacute;cimo\u003c/p\u003e\u003cp\u003eGu\u0026aacute;piles\u003c/p\u003e\u003cp\u003eRita\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eH67*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eAB907796.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eCosta Rica\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e1390/1390\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"6\"\u003e* Haplotypes previously reported in Costa Rica.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003ePolymorphic sites of the haplotypes of \u003cem\u003eAe. aegypti\u003c/em\u003e and \u003cem\u003eAe. albopictus\u003c/em\u003e from Costa Rica found in the sequences of the COI gene.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"16\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c13\" colnum=\"13\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c14\" colnum=\"14\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c15\" colnum=\"15\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c16\" colnum=\"16\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eAe. aegypti\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"15\" nameend=\"c16\" namest=\"c2\"\u003e\u003cp\u003ePolymorphism position\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eHaplotype\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e33\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e168\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e234\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e237\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003e246\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cb\u003e318\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u003cb\u003e330\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e\u003cb\u003e351\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e435\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e\u003cb\u003e480\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c12\"\u003e\u003cp\u003e\u003cb\u003e501\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c13\"\u003e\u003cp\u003e\u003cb\u003e579\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c14\"\u003e\u003cp\u003e\u003cb\u003e618\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c15\"\u003e\u003cp\u003e\u003cb\u003e630\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c16\"\u003e\u003cp\u003e\u003cb\u003e636\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eH1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003eG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003eG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c12\"\u003e\u003cp\u003eC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c13\"\u003e\u003cp\u003eA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c14\"\u003e\u003cp\u003eT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c15\"\u003e\u003cp\u003eG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c16\"\u003e\u003cp\u003eC\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eH2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003eA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003eA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c12\"\u003e\u003cp\u003eC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c13\"\u003e\u003cp\u003eA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c14\"\u003e\u003cp\u003eC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c15\"\u003e\u003cp\u003eG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c16\"\u003e\u003cp\u003eC\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eH3\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003eG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003eA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c12\"\u003e\u003cp\u003eT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c13\"\u003e\u003cp\u003eG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c14\"\u003e\u003cp\u003eT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c15\"\u003e\u003cp\u003eA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c16\"\u003e\u003cp\u003eT\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eAe. albopictus\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"6\" nameend=\"c7\" namest=\"c2\"\u003e\u003cp\u003e\u003cb\u003ePolymorphism position\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"9\" nameend=\"c16\" namest=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eHaplotype\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e40\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e94\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e467\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e954\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003e1140\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cb\u003e1404\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"9\" nameend=\"c16\" namest=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eH3\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"9\" nameend=\"c16\" namest=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eH6\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"9\" nameend=\"c16\" namest=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eH26\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"9\" nameend=\"c16\" namest=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eH37\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"9\" nameend=\"c16\" namest=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eH67\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"9\" nameend=\"c16\" namest=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003ePolymorphisms with amino acid changes in different haplotypes of \u003cem\u003eAe. aegypti\u003c/em\u003e and \u003cem\u003eAe. albopictus\u003c/em\u003e from Costa Rica according to the COI gene sequence.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"4\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eAe. aegypti\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"3\" nameend=\"c4\" namest=\"c2\"\u003e\u003cp\u003eChange position in amino acids\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eHaplotype\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e168\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e246\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e579\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eH1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eMethionine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMethionine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eIsoleucine\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eH2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eIsoleucine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMethionine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eIsoleucine\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eH3\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eIsoleucine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eIsoleucine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eMethionine\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eAe. albopictus\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"3\" nameend=\"c4\" namest=\"c2\"\u003e\u003cp\u003e\u003cb\u003eChange position in amino acids\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eHaplotype\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e40\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e94\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eH3\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eIsoleucine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eValine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eH6\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePhenylalanine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eValine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eH26\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eIsoleucine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eValine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eH37\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eIsoleucine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eValine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eH67\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eIsoleucine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eIsoleucine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eThe COI gene sequences of H1, H2, and H4 haplotypes of \u003cem\u003eAe. aegypti\u003c/em\u003e showed four, two, and nine polymorphic sites, respectively (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA), with the amino acid sequence affecting the polymorphic sites at positions 168, 246, and 579. At position 168, H1 encoded a methionine, whereas H2 and H4 encoded isoleucine. At position 246, H1 and H2 encoded methionine, and H4 isoleucine; finally, at position 579, H1 and H2 encoded isoleucine, and H4 methionine (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003eA). The H3, H26, H37 and H67 haplotypes of \u003cem\u003eAe. albopictus\u003c/em\u003e presented a polymorphic site in their sequences, whereas the H6 haplotype presented two polymorphic sites (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003eB). The amino acid sequence of the polymorphic sites was established at positions 40 and 94. At position 40, H6 encoded a phenylalanine, while the rest of the haplotypes encoded an isoleucine, and at position 94, H67 encoded an isoleucine, while the rest encoded valine (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003eB). The phylogenetic tree based on the COI gene established three well-defined clades for \u003cem\u003eAe. aegypti\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e) and five well-defined clades for \u003cem\u003eAe. albopictus\u003c/em\u003e, presenting a clade for each haplotype (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe number of haplotypes of \u003cem\u003eAe. aegypti\u003c/em\u003e (n\u0026thinsp;=\u0026thinsp;8) determined using a total of 18 larvae collected from 18 locations in Costa Rica was similar to that reported in El Salvador (10 haplotypes in a total of 84 samples) and Panama (13 haplotypes in a total of 122 samples), and these larvae were collected from six and 30 locations, respectively [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. Compared with these reports, numerous \u003cem\u003eAe. aegypti\u003c/em\u003e larvae with different haplotypes were detected, although only one larva of each species was analyzed in each district due to economic limitations. In \u003cem\u003eAe. albopictus\u003c/em\u003e, the number of haplotypes detected in a small number of larvae (5 haplotypes in 11 larvae) was similar to that reported in Brazil (10 haplotypes in 163 samples from 14 localities) [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e] and in Colombia (33 haplotypes in 163 samples and 87 haplotypes in 145 samples from 14 and six locations, respectively) [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. This could be due to the climatic and geographical similarities between Costa Rica and the countries of Central and South America; if more larvae are collected in different regions of Costa Rica, greater diversity may be found [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eIn this study, there are two haplotypes of \u003cem\u003eAe. aegypti\u003c/em\u003e (H1 and H2) and three haplotypes of \u003cem\u003eAe. albopictus\u003c/em\u003e (H3, H6, and H26) are reported for the first time in Costa Rica. Each of these haplotypes was detected in only one district, except for haplotype 1 of \u003cem\u003eAe. aegypti\u003c/em\u003e, which was widely distributed in a total of 14 districts [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. This haplotype has also been reported in El Salvador, where it is the most widely distributed haplotype in the country [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. Likewise, in Honduras, it is the most abundant and widespread haplotype [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. Haplotype 2 of \u003cem\u003eAe. aegypti\u003c/em\u003e was found only in the La Isabel district in the southern central region of the country and had been previously reported in Panama [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. Haplotype 3 of \u003cem\u003eAe. aegypti\u003c/em\u003e was found in three districts (Pital, Siquirres, and Turrialba) and has already been reported previously in the Siquirres district [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. In contrast with haplotypes 1 and 2 of \u003cem\u003eAe. aegypti\u003c/em\u003e, which have only been reported to date in El Salvador, Honduras, and Panama, haplotype 4 has also been found in countries such as Canada, New Caledonia, and Germany, in addition to Central America.\u003c/p\u003e\u003cp\u003eHaplotypes 3 and 6 of \u003cem\u003eAe. albopictus\u003c/em\u003e were found in the southern districts of the country and have been previously reported only in Japan, Italy, USA (H3), and China (H6). The third newly discovered haplotype (H26) of \u003cem\u003eAe. albopictus\u003c/em\u003e was found in the northern districts of the country and has been previously reported in Singapore. The other two haplotypes detected (H37 and H67) had previously been found in the district of La Virgen de Sarapiqu\u0026iacute; [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. In the present study, H37 was found to be distributed in the Pacific and Atlantic areas of the country, whereas H67 was found to be more widely distributed, but only in the Atlantic area. Our findings agree with those of Futami et al. [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e], who reported a high diversity of haplotypes of \u003cem\u003eAe. albopictus\u003c/em\u003e (7 haplotypes) in La Virgen de Sarapiqu\u0026iacute;. We found an additional haplotype in the same district, reporting for the first time the presence of H26 in La Virgen. In studies carried out in Medell\u0026iacute;n, Colombia, different haplotypes were determined in three neighborhoods and two time periods (January\u0026ndash;April 2010 and October\u0026ndash;December 2012). The results revealed that the genetic composition changes occurred without a predictable pattern in 2 years [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e].\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe genetic diversity of \u003cem\u003eAedes\u003c/em\u003e populations in Costa Rica was confirmed, and previously unreported haplotypes were reported. Seasonal and geographically broader studies are recommended to understand vector population dynamics. For example, conducting systematic studies with multiple larvae from each district in the dry and rainy seasons, to establish seasonal differences, which could increase our understanding of genetic diversity, reproduction speed, and genetic flow of natural mosquito populations. These analyses would provide not only an evolutionary perspective on vector populations but also a key tool for designing and optimizing control strategies that can slow the spread of diseases transmitted by arbovirus vectors.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe Costa Rican Ministry of Health, especially Rodrigo Mar\u0026iacute;n, Manuel Guti\u0026eacute;rrez, Gabriela Delgado, Carlos Aguilar, and the regional technicians who assisted with sample collection. The German Academic Exchange Service (DAAD) awarded scholarships to Sonia Flores N\u0026uacute;\u0026ntilde;ez and Edwin Beltr\u0026eacute; Nu\u0026ntilde;ez.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe project was funded by FUNDAUNA \u0026ldquo;Diagnosis and Research in Population and Ecosystem Medicine\u0026rdquo; and Vicerrector\u0026iacute;a de Investigaci\u0026oacute;n, Universidad Nacional.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data sets used during the current study are available from the corresponding authors on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eEach author\u0026apos;s contribution to the manuscript is as follows:\u003c/p\u003e\n\u003cp\u003eEdwin Beltr\u0026eacute;-Nu\u0026ntilde;ez: conceptualization, methodology, formal analysis, investigation, data curation, writing - original draft.\u003c/p\u003e\n\u003cp\u003eSonia Flores-Nu\u0026ntilde;ez: conceptualization, methodology, formal analysis, investigation, data curation, writing - original draft.\u003c/p\u003e\n\u003cp\u003eSilvia Arg\u0026uuml;ello-Vargas: conceptualization, methodology, investigation, resources, writing - original draft, writing - review, supervision, project administration, funding.\u003c/p\u003e\n\u003cp\u003eAntony Solorzano-Morales: methodology, investigation, writing - review, editing.\u003c/p\u003e\n\u003cp\u003eGaby Dolz: conceptualization, methodology, investigation, resources, writing -\u003c/p\u003e\n\u003cp\u003eoriginal draft, writing - review, supervision, project administration, funding.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate, consent for publication, competing interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have fully complied with all relevant ethical and legal requirements, both during the study and in the production of the manuscript; that there are no conflicts of interest of any kind; that all funding sources are fully and clearly stated in the acknowledgments section; and that they fully agree with the final edited version of the article. 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Regi\u0026oacute;n Huetar Norte: oferta exportada actual y oferta potencial de productos agropecuarios alternativos (No. E71/10543). Promotora del Comercio Exterior de Costa Rica. 2008. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://www.mag.go.cr/bibliotecavirtual/E71-10543.pdf\u003c/span\u003e\u003cspan address=\"http://www.mag.go.cr/bibliotecavirtual/E71-10543.pdf\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Mosquitos, vectors, phylogenetics, polymorphisms, clades","lastPublishedDoi":"10.21203/rs.3.rs-7255371/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7255371/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e\u003cp\u003e\u003cem\u003eAedes aegypti\u003c/em\u003e and \u003cem\u003eAedes albopictus\u003c/em\u003e are key vectors of arboviruses such as dengue, chikungunya, and Zika. Monitoring the genetic diversity of these mosquito species is essential for understanding their adaptation, distribution, and implications for disease control. This study aimed to identify and compare the haplotypes of \u003cem\u003eAedes aegypti\u003c/em\u003e and \u003cem\u003eAedes albopictus\u003c/em\u003e larvae in 18 districts of Costa Rica.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e\u003cp\u003eA cross-sectional, descriptive study was conducted from August to November 2021. Larvae were collected from 26 locations across 18 districts at elevations below 900 m above sea level. Morphological identification was followed by DNA extraction and amplification of the cytochrome oxidase I (COI) mitochondrial gene by Polymerase Chain Reaction. Sequences were aligned and compared with GenBank references. Phylogenetic analysis was conducted using the unweighted pair-group method with arithmetic averages (UPGMA) clustering method.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e\u003cp\u003e\u003cem\u003eAe. aegypti\u003c/em\u003e was detected in all districts, and \u003cem\u003eAedes albopictus\u003c/em\u003e in 11. Three haplotypes of \u003cem\u003eAe. aegypti\u003c/em\u003e (H1, H2, H4) and five of \u003cem\u003eAe. albopictus\u003c/em\u003e (H3, H6, H26, H37, H67) were identified. H1 (\u003cem\u003eAe. aegypti\u003c/em\u003e) and H67 (\u003cem\u003eAe. albopictus\u003c/em\u003e) were the most widespread. Several haplotypes not previously reported in Costa Rica were found. Amino acid polymorphisms were noted at key COI positions, with distinct clades observed in phylogenetic trees.\u003c/p\u003e\u003ch2\u003eConclusions\u003c/h2\u003e\u003cp\u003eThis study confirms the genetic diversity of \u003cem\u003eAedes\u003c/em\u003e populations in Costa Rica and identifies previously unreported haplotypes. Further seasonal and geographically broader studies are recommended to track vector population dynamics and inform vector control strategies.\u003c/p\u003e","manuscriptTitle":"Haplotypes of Aedes aegypti and Aedes albopictus larvae detected in 18 districts of Costa Rica","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-08-27 17:06:52","doi":"10.21203/rs.3.rs-7255371/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"a82b2b72-80f7-48e1-a52c-9c6ba0802857","owner":[],"postedDate":"August 27th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-11-05T16:40:25+00:00","versionOfRecord":[],"versionCreatedAt":"2025-08-27 17:06:52","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7255371","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7255371","identity":"rs-7255371","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}