• 1.

    Mackenzie J, Gubler D, Petersen L, 2004. Emerging flaviviruses: the spread and resurgence of Japanese encephalitis, West Nile and dengue viruses. Nat Med 10: S98S109.

    • Search Google Scholar
    • Export Citation
  • 2.

    World Health Organization, 2013. Sustaining the Drive to Overcome the Global Impact of Neglected Tropical Diseases. Second WHO Report on Neglected Diseases. Geneva: WHO Press, 2529.

    • Search Google Scholar
    • Export Citation
  • 3.

    Weaver S, Vasilakis N, 2009. Molecular evolution of dengue viruses: contributions of phylogenetics to understand the history and epidemiology of the preeminent arboviral disease. Infect Genet Evol 9: 523540.

    • Search Google Scholar
    • Export Citation
  • 4.

    World Health Organization, 2009. Dengue: Guidelines for Diagnosis, Treatment, Prevention and Control. Geneva: WHO Press, 324.

  • 5.

    Vasilakis N, Cardosa J, Hanley K, Holmes E, Weaver S, 2011. Fever from the forest: prospects for the continued emergence of sylvatic dengue virus and its impact on public health. Nat Rev Microbiol 9: 532541.

    • Search Google Scholar
    • Export Citation
  • 6.

    Roberts D, Peyton E, Pinheiro FP, Balderrama F, Vargas R, 1984. Associations of arbovirus vectors with gallery forest and domestic environments in southeastern Bolivia. Bull Pan Am Health Organ 18: 337350.

    • Search Google Scholar
    • Export Citation
  • 7.

    De Thoisy B, Lacoste V, Germain A, Muñoz-Jordán J, Colón C, Mauffrey JF, Delaval M, Catzeflis F, Kasanji M, Matheus S, Dussart P, Morvan J, Aguilar-Setien A, Deparis X, Lavergne A, 2009. Dengue infection in neotropical forest mammals. Vector Borne Zoonotic Dis 9: 157169.

    • Search Google Scholar
    • Export Citation
  • 8.

    Calisher C, Holmes K, Domínguez S, Shountz T, Cryan P, 2008. Bats prove to be rich reservoirs for emerging viruses. Microbe Wash DC 3: 521528.

  • 9.

    Turmelle A, Olival K, 2009. Correlates of viral richness in bats (order Chiroptera). EcoHealth 6: 522539.

  • 10.

    Drexler JF, Corman VM, Müller MA, Maganga GD, Vallo P, Binger T, Gloza-Rausch F, Cottontail VM, Rasche A, Yordanov S, Seebens A, Knornschild M, Oppong S, Adu Sarkodie Y, Pongombo C, Lukashev AN, Schmidt-Chanasit J, Stöcker A, Carneiro AJ, Erbar S, Maisnes A, Fornhoffs F, Buettner R, Kalko EK, Kruppa T, Franke CR, Kallies R, Yandoko ER, Herrler G, Reusken C, Hassanin A, Krüger DH, Matthee S, Ulrich RG, Leroy EM, Drosten C, 2012. Bat host major mammalian paramyxoviruses. Nat Commun 3: 796.

    • Search Google Scholar
    • Export Citation
  • 11.

    Olival K, Epstein JH, Wang LF, Field H, Daszak P, 2012. Are bats exceptional viral reservoirs? Aguirre A, Ostfeld R, Daszak P, eds. New Directions in Conservation Medicine: Applied Cases of Ecological Health, 1st Ed. New York, NY: Oxford University Press, 195212.

    • Search Google Scholar
    • Export Citation
  • 12.

    Anthony S, Ojeda-Flores R, Rico-Chávez O, Navarrete-Macias I, Zambrana-Torrelio C, Rostal MK, Epstein JH, Tipps T, Liang E, Sanchez-Leon M, Sotomayor-Bonilla J, Aguirre AA, Ávila-Flores R, Medellín RA, Goldstein T, Suzán G, Daszak P, Lipkin WI, 2013. Coronavirus in bats from Mexico. J Gen Virol 94: 10281038.

    • Search Google Scholar
    • Export Citation
  • 13.

    O'Connor J, Rowan L, Lawrence J, 1955. Relationships between the flying fox (genus Pteropus) and arthropod-borne fevers of North Queensland. Nature 176: 472.

    • Search Google Scholar
    • Export Citation
  • 14.

    Zhang H, Yang X, Li G: Detection of dengue virus genome RNA in some kinds of animal caught from dengue fever endemic areas in Hainan Island with reverse transcription-polymerase chain reaction. Zhang Shi Yan He Lin Chuang Du Xue Za Zhi 12: 226228.

    • Search Google Scholar
    • Export Citation
  • 15.

    Platt K, Mangiafico J, Rocha O, Zaldivar M, Mora J, Trueba G, Rowley WA, 2000. Detection of dengue virus neutralizing antibodies in bats from Costa Rica and Ecuador. J Med Entomol 37: 965967.

    • Search Google Scholar
    • Export Citation
  • 16.

    Aguilar-Setien A, Romero-Almaraz M, Sánchez-Hernández C, Figueroa R, Juárez-Palma LP, García-Flores MM, Vásquez-Salinas C, Salas-Rojas M, Hidalgo-Martínez AC, Aguilar-Pierlé S, García-Estrada C, Ramos C, 2008. Dengue virus in Mexican bats. Epidemiol Infect 136: 16781683.

    • Search Google Scholar
    • Export Citation
  • 17.

    Machain-Williams C, López-Uribe M, Talavera-Aguilar L, Carrillo-Navarrete J, Vera-Escalante L, Puerto-Manzano F, Ulloa A, Farfán-Ale JA, García-Rejón J, Blitvich BJ, Loroño-Pino MA, 2013. Serologic evidence of flavivirus infection in bats in the Yucatan Peninsula of Mexico. J Wildl Dis 49: 684689.

    • Search Google Scholar
    • Export Citation
  • 18.

    Medellín R, Arita H, Sánchez O, 2008. Identificación de los Murciélagos de México. Clave de Identificación de Campo. Mexico City, México: Instituto de Ecología, UNAM, 2878.

    • Search Google Scholar
    • Export Citation
  • 19.

    Sikes RS, Gannon WL; ACUCASM, 2011. Guidelines of the American Society of Mammalogists for the use of wild mammals in research. J Mammal 92: 235253.

    • Search Google Scholar
    • Export Citation
  • 20.

    Lanciotti R, Calisher C, Gubler D, Chang G, Vorndam AV, 1992. Rapid detection and typing of dengue viruses from clinical samples by using reverse transcriptase-polymerase chain reaction. J Clin Microbiol 30: 545551.

    • Search Google Scholar
    • Export Citation
  • 21.

    Medellín RA, Equihua M, Amin M, 2000. Bat diversity and abundance as indicators of disturbance in neotropical rainforest. Conserv Biol 14: 16661675.

    • Search Google Scholar
    • Export Citation
  • 22.

    Vargas J, Medellín RA, Escalona-Segura G, Interián-Sosa L, 2009. Vegetation complexity and bat-plant dispersal in Calakmul, Mexico. J Nat Hist 43: 219243.

    • Search Google Scholar
    • Export Citation
  • 23.

    Estrada A, Coates-Estrada R, Meritt D, 1993. Bat species richness and abundance in a tropical rain forest fragments and in agricultural habitats at Los Tuxtlas, Mexico. Ecography (Cop.) 16: 309318.

    • Search Google Scholar
    • Export Citation
  • 24.

    R Core Team, 2013. R: A Language and Environment for Statistical Computing. Vienna, Austria: R Foundation for Statistical Computing.

  • 25.

    Gómez Dantes H, 2007. Elementos económicos y políticos que impactan en el control del dengue en México. Salud Publica Mex 49: 117119.

    • Search Google Scholar
    • Export Citation

 

 

 

 

 

Dengue Virus in Bats from Southeastern Mexico

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  • Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México, México, Distrito Federal, México; Escuela de Biología, Universidad de Costa Rica, San José, Costa Rica; EcoHealth Alliance, New York, New York; Unidad de Investigación Médica en Inmunología, Coordinación de Investigación en Salud, Instituto Mexicano del Seguro Social, México, Distrito Federal, México; Red Ambiente y Sustentabilidad, Instituto de Ecología AC, Veracruz, México; Laboratorio de Medicina de Conservación, Escuela Superior de Medicina, Instituto Politécnico Nacional, México, Distrito Federal, México; Department of Environmental Science and Policy, George Mason University, Fairfax, Virginia; Smithsonian–Mason School of Conservation, Front Royal, Virginia

To identify the relationship between landscape use and dengue virus (DENV) occurrence in bats, we investigated the presence of DENV from anthropogenically changed and unaltered landscapes in two Biosphere Reserves: Calakmul (Campeche) and Montes Azules (Chiapas) in southern Mexico. Spleen samples of 146 bats, belonging to 16 species, were tested for four DENV serotypes with standard reverse transcriptase polymerase chain reaction (RT-PCR) protocols. Six bats (4.1%) tested positive for DENV-2: four bats in Calakmul (two Glossophaga soricina, one Artibeus jamaicensis, and one A. lituratus) and two bats in Montes Azules (both A. lituratus). No effect of anthropogenic disturbance on the occurrence of DENV was detected; however, all three RT-PCR–positive bat species are considered abundant species in the Neotropics and well-adapted to disturbed habitats. To our knowledge, this study is the first study conducted in southeastern Mexico to identify DENV-2 in bats by a widely accepted RT-PCR protocol. The role that bats play on DENV's ecology remains undetermined.

Dengue fever is an important public health concern in the tropics,14 and ecological and epidemiological studies are needed to assess the role of bats and other mammals in a possible sylvatic maintenance cycle.5 Dengue viruses (DENVs) comprise four antigenically distinct but genetically related serotypes of the Flavivirus genus (Flaviviridae family).1 DENVs are positive-sense single-stranded RNA viruses that cause one of the most common infectious diseases in humans in tropical regions.2 Their transmission includes an urban endemic/epidemic cycle between Aedes aegypti mosquitoes and humans as the reservoir host and a sylvatic enzootic cycle between non-human primates and arboreal mosquitoes of the genus Aedes.3 The urban cycle is well-documented in the Neotropics, with four serotypes reported in urban areas,14 whereas the sylvatic cycle has been shown in West Africa and peninsular Malaysia.5 Thus far, the sylvatic cycle has not been described in the Neotropics. However, in Bolivia, DENV seroconversions among the indigenous Ayoreo people were found in a remote area where Ae. aegypti, the primary vector, was absent.6 This finding suggests a possible sylvatic cycle involving a different mosquito species or cross-reaction with antibodies to another flavivirus. In French Guiana, all four DENV serotypes have been identified by molecular methods in 92 wild mammals (bats, rodents, and marsupials) in all settings investigated: periurban, rural, and sparsely populated areas.7 This finding suggests that primarily urban DENV strains could infect wildlife in non-urban forested areas.7 The role of wildlife in DENV transmission remains unknown.

Bats are important reservoirs of many viruses, such as rabies viruses, Nipah viruses, and coronaviruses.812 Flaviviridae are the second most frequently reported viral family in the order Chiroptera (13% frequency; second only to rhabdoviruses)9; however, their role in the dynamics of DENVs remains poorly understood. DENV have been reported in large frugivorous bats (Pteropus genus) in Australia.13 More recently, polymerase chain reaction (PCR) and antigen detection (direct immunofluorescent assay) of DENV were described in bats from DENV-endemic islands in China.14 Seroconversion in bats (DENV-1, -2, and -3) has been reported in urban areas in Costa Rica and Ecuador.15 Bats captured during a dengue fever outbreak along the Gulf and Pacific Coasts of Mexico were found to be DENV-seropositive (enzyme-linked immunosorbent assay [ELISA]), have DENV NS1 protein, and be positive on reverse transcriptase (RT) -PCR for DENV-2.16 Molecular evidence of DENV-1, -2, and -3 in bats from French Guiana was reported, including a phylogenetic sequence from a Carollia perspicillata bat consistent with DENV-1.7 Qualitative detection of the NS1 antigen of DENVs in bat serum samples and plaque reduction neutralization tests has been reported in bats from southeastern Mexico from disturbed sites near human settlements in Campeche (S. Cabrera-Romo and others, unpublished data) and Mérida,17 which is the largest city in the Yucatan Peninsula.

In this study, samples were collected from bats trapped in two biosphere reserves located in southeastern Mexico. High bat diversity and large tracts of continuous forest characterize these areas. Additionally, DENV-1, -2, and -4 have been reported in human populations. The aim of this study was to show the presence of DENV serotypes in bats within the biosphere reserves and adjacent areas with anthropogenic changes using RT-PCR. Bats were collected in Montes Azules Biosphere Reserve (Chiapas; 16°9′46″ N, 90°41′18″ W) and Calakmul Biosphere Reserve (Campeche; 18°26′1″ N, 89°36′61″ W) from two landscape types categorized as undisturbed forest (UD), comprised of primary forest with no human disturbance, and disturbed forest (D), defined as a transitional zone between primary forest and agricultural/livestock areas or human settlements. In both regions, the sites were located at least 20 km from significant human populations where dengue has been reported (Xpujil in Campeche State and Benemérito de la Américas in Chiapas State). Bats were captured using four mist nets (5 × 9 m) in foraging sites. Nets were opened at dusk and remained open for 4 consecutive hours. Each site was sampled three times between November of 2010 and August of 2011. Field guides were used for the taxonomic species identification of all bats captured.18

In total, 146 bats were euthanized following the Guidelines of the American Society of Mammalogists for the Use of Wild Mammals in Research19 with the approval of the Institutional Animal Care and Use Subcommittee of the Veterinary Faculty of Universidad Nacional Autónoma de México. One hundred forty-six spleen specimens were collected and preserved in liquid nitrogen, and RNA from all samples was extracted using TRIzol LS Reagent (Thermo Fisher Scientific Inc., Carlsbad, CA) according to the manufacturer's instructions. RT-PCR was carried out for DENV (DENV-1, -2, -3, and -4) as described previously using highly specific and sensitive primers.20 The DENV-positive control strains used in the RT-PCR for DENVs (D1/AO/XX/1988 Angola, D2/CR/13538/2007 Limón, D3/CR/14532/2007 Corredores, and D4/DM/814669/1981 Dominica) were donated by the Dengue Reference Center, Instituto Costarricense de Investigación y Enseñanza en Nutrición y Salud (INCIENSA; Costa Rica). Nuclease-free water was used as a negative control. RT-PCR products were visualized by 2% agarose gel electrophoresis and stained with Gel Red (Biotium Inc., Hayward, CA). Positive and negative controls resulted as expected.

Samples from 16 bat species were analyzed. Six DENV-2–positive bats were detected (prevalence of 4.1%; 95% confidence interval ± 3.22) (Table 1). In Calakmul, four bats were PCR-positive: two Glossophaga soricina, one Artibeus jamaicensis, and one A. lituratus. Two bats, G. soricina and A. jamaicensis, were captured in the UD sites, and two bats, G. soricina and A. lituratus, were captured in a D site. In Montes Azules, two A. lituratus were positive to DENV-2: one bat from a UD site and one bat from a D site. These three bat species (A. jamaicensis, A. lituratus, and G. soricina) are considered highly abundant bat species in the Mexican Neotropics,2123 coinciding with the relative abundance found in this study (A. lituratus: 179 [22%]; G. soricina: 118 [14.5%]; and A. jamaicensis: 111 [13.6%]). No significant difference was detected comparing the presence of DENV-2 and the type of site (UD and D) using a test of equal or given proportions in R242 = 0.1738, degrees of freedom = 1, P = 0.6768).

Table 1

Number of bats sampled by species by site type collected and analyzed for DENV between 2010 and 2011 from Calakmul and Montes Azules

Bat speciesCalakmulMontes Azules
UDDUDD
Artibeus jamaicensis14*87 
A. lituratus216*13*7*
A. phaeotis1   
Carollia perspicillata1   
C. sowelli  311
Desmodus rotundus   1
Glossophaga commissarisi1  1
G. soricina7*22*24
Hylonycteris underwoodi  1 
Platyrrhinus helleri   1
Pteronotus davyi1   
P. parnelli51  
S. bilineata   1
Sturnira lilium 624
S. ludovici  11
Uroderma bilobatum   1
Total32532932

A positive PCR result for DENV-2.

This preliminary study provides additional evidence of DENV exposure in Neotropical bats.7,1517 Unfortunately, because of sample degradation, we could not amplify a different region of the viral genome for phylogenetic analysis from the positive samples. Although DENV infection was serologically recognized in 26 bats of the same three species (A. jamaicensis, A. lituratus, and G. soricina) from Merida City,17 this detection is the first detection of the DENV-2 genome by molecular diagnostic methods in bats from southeastern Mexico. DENV-1, -2, and -3 have been reported in bats from French Guiana by RT-PCR, where DENV was found in A. planirostris and C. perspicillata.7 Along the Gulf Coast of Mexico, DENV-2 was identified in Myotis nigricans, Carollia sowelli, and A. jamaicensis.16 However, this detection is the first molecular detection of the DENV-2 genome in A. lituratus and G. soricina.

Old World DENV-2 strains diverged relatively recently and spread to the Neotropics by human carriers and the transportation of infected monkeys and mosquitoes.5 The encroachment of human settlements and agricultural areas on UD areas could promote new interactions among DENV vectors and potential hosts, increasing the possible development of sylvatic DENV.5 Large-scale research of anthropogenic change and pathogen evolution using an epidemiologic framework is vital to develop comprehensive policies across public health, economic development, and conservation biology.5

This study adds additional evidence for the presence of DENV-2 in three bat species (A. jamaicensis, A. lituratus, and G. soricina). However, the extent to which bats are involved in DENV transmission and their ability to act as competent hosts remain undetermined. Because these species are the three most abundant species in the region, we may be seeing random spillover into dead-end hosts. Understanding the ecology of DENV in bats is important before implicating bats as a threat to public health. Sylvatic DENV in any species could evolve from biological interactions between wild reservoirs and sylvatic vectors.5,7 Lastly, it may arise from socioeconomic factors, such as human encroachment into natural areas in locations where medical and health services are absent.25 Future investigations of DENV in Neotropical bats and other potential wild reservoirs will improve the understanding of the ecological dynamics of these viruses in nature.

ACKNOWLEDGMENTS

We acknowledge the Posgrado en Ciencias de la Producción y Salud Animal of the Universidad Nacional Autonóma de México (UNAM) and the Consejo Nacional de Ciencia y Tecnología (CONACYT) for supporting this research. We thank Annkatrin Junglass, associate public relations manager of Qiagen, Germany, for donating diagnostic supplies. We are grateful to Carolyn Brown and Kendra Shannon from UNAM–Canada for reviewing this manuscript. In addition, we thank the Comisión Nacional de Áreas Protegidas (CONANP) de Xpujil, Campeche, especially José A. Zuñiga and Francisco Pérez, for facilitating the field work in Calakmul. Also, we thank Rafael Lombera and his family from Chajul, Chiapas for help with the fieldwork. We are grateful to Angélica Menchaca, Paola Martínez, Karen Moreno, Adriana Fernández, Shiara Gonzalez, Luis R. Viquez, Fernando Salgado, Rosa Tenorio, Amanda Vicente, and Leonardo Perea. Finally, thanks to the Production and Animal Health Sciences Graduate Program and special thanks to the Disease Ecology Group from the Ethology, Wildlife and Laboratory Animals Department, Veterinary Faculty (FMVZ), UNAM.

  • 1.

    Mackenzie J, Gubler D, Petersen L, 2004. Emerging flaviviruses: the spread and resurgence of Japanese encephalitis, West Nile and dengue viruses. Nat Med 10: S98S109.

    • Search Google Scholar
    • Export Citation
  • 2.

    World Health Organization, 2013. Sustaining the Drive to Overcome the Global Impact of Neglected Tropical Diseases. Second WHO Report on Neglected Diseases. Geneva: WHO Press, 2529.

    • Search Google Scholar
    • Export Citation
  • 3.

    Weaver S, Vasilakis N, 2009. Molecular evolution of dengue viruses: contributions of phylogenetics to understand the history and epidemiology of the preeminent arboviral disease. Infect Genet Evol 9: 523540.

    • Search Google Scholar
    • Export Citation
  • 4.

    World Health Organization, 2009. Dengue: Guidelines for Diagnosis, Treatment, Prevention and Control. Geneva: WHO Press, 324.

  • 5.

    Vasilakis N, Cardosa J, Hanley K, Holmes E, Weaver S, 2011. Fever from the forest: prospects for the continued emergence of sylvatic dengue virus and its impact on public health. Nat Rev Microbiol 9: 532541.

    • Search Google Scholar
    • Export Citation
  • 6.

    Roberts D, Peyton E, Pinheiro FP, Balderrama F, Vargas R, 1984. Associations of arbovirus vectors with gallery forest and domestic environments in southeastern Bolivia. Bull Pan Am Health Organ 18: 337350.

    • Search Google Scholar
    • Export Citation
  • 7.

    De Thoisy B, Lacoste V, Germain A, Muñoz-Jordán J, Colón C, Mauffrey JF, Delaval M, Catzeflis F, Kasanji M, Matheus S, Dussart P, Morvan J, Aguilar-Setien A, Deparis X, Lavergne A, 2009. Dengue infection in neotropical forest mammals. Vector Borne Zoonotic Dis 9: 157169.

    • Search Google Scholar
    • Export Citation
  • 8.

    Calisher C, Holmes K, Domínguez S, Shountz T, Cryan P, 2008. Bats prove to be rich reservoirs for emerging viruses. Microbe Wash DC 3: 521528.

  • 9.

    Turmelle A, Olival K, 2009. Correlates of viral richness in bats (order Chiroptera). EcoHealth 6: 522539.

  • 10.

    Drexler JF, Corman VM, Müller MA, Maganga GD, Vallo P, Binger T, Gloza-Rausch F, Cottontail VM, Rasche A, Yordanov S, Seebens A, Knornschild M, Oppong S, Adu Sarkodie Y, Pongombo C, Lukashev AN, Schmidt-Chanasit J, Stöcker A, Carneiro AJ, Erbar S, Maisnes A, Fornhoffs F, Buettner R, Kalko EK, Kruppa T, Franke CR, Kallies R, Yandoko ER, Herrler G, Reusken C, Hassanin A, Krüger DH, Matthee S, Ulrich RG, Leroy EM, Drosten C, 2012. Bat host major mammalian paramyxoviruses. Nat Commun 3: 796.

    • Search Google Scholar
    • Export Citation
  • 11.

    Olival K, Epstein JH, Wang LF, Field H, Daszak P, 2012. Are bats exceptional viral reservoirs? Aguirre A, Ostfeld R, Daszak P, eds. New Directions in Conservation Medicine: Applied Cases of Ecological Health, 1st Ed. New York, NY: Oxford University Press, 195212.

    • Search Google Scholar
    • Export Citation
  • 12.

    Anthony S, Ojeda-Flores R, Rico-Chávez O, Navarrete-Macias I, Zambrana-Torrelio C, Rostal MK, Epstein JH, Tipps T, Liang E, Sanchez-Leon M, Sotomayor-Bonilla J, Aguirre AA, Ávila-Flores R, Medellín RA, Goldstein T, Suzán G, Daszak P, Lipkin WI, 2013. Coronavirus in bats from Mexico. J Gen Virol 94: 10281038.

    • Search Google Scholar
    • Export Citation
  • 13.

    O'Connor J, Rowan L, Lawrence J, 1955. Relationships between the flying fox (genus Pteropus) and arthropod-borne fevers of North Queensland. Nature 176: 472.

    • Search Google Scholar
    • Export Citation
  • 14.

    Zhang H, Yang X, Li G: Detection of dengue virus genome RNA in some kinds of animal caught from dengue fever endemic areas in Hainan Island with reverse transcription-polymerase chain reaction. Zhang Shi Yan He Lin Chuang Du Xue Za Zhi 12: 226228.

    • Search Google Scholar
    • Export Citation
  • 15.

    Platt K, Mangiafico J, Rocha O, Zaldivar M, Mora J, Trueba G, Rowley WA, 2000. Detection of dengue virus neutralizing antibodies in bats from Costa Rica and Ecuador. J Med Entomol 37: 965967.

    • Search Google Scholar
    • Export Citation
  • 16.

    Aguilar-Setien A, Romero-Almaraz M, Sánchez-Hernández C, Figueroa R, Juárez-Palma LP, García-Flores MM, Vásquez-Salinas C, Salas-Rojas M, Hidalgo-Martínez AC, Aguilar-Pierlé S, García-Estrada C, Ramos C, 2008. Dengue virus in Mexican bats. Epidemiol Infect 136: 16781683.

    • Search Google Scholar
    • Export Citation
  • 17.

    Machain-Williams C, López-Uribe M, Talavera-Aguilar L, Carrillo-Navarrete J, Vera-Escalante L, Puerto-Manzano F, Ulloa A, Farfán-Ale JA, García-Rejón J, Blitvich BJ, Loroño-Pino MA, 2013. Serologic evidence of flavivirus infection in bats in the Yucatan Peninsula of Mexico. J Wildl Dis 49: 684689.

    • Search Google Scholar
    • Export Citation
  • 18.

    Medellín R, Arita H, Sánchez O, 2008. Identificación de los Murciélagos de México. Clave de Identificación de Campo. Mexico City, México: Instituto de Ecología, UNAM, 2878.

    • Search Google Scholar
    • Export Citation
  • 19.

    Sikes RS, Gannon WL; ACUCASM, 2011. Guidelines of the American Society of Mammalogists for the use of wild mammals in research. J Mammal 92: 235253.

    • Search Google Scholar
    • Export Citation
  • 20.

    Lanciotti R, Calisher C, Gubler D, Chang G, Vorndam AV, 1992. Rapid detection and typing of dengue viruses from clinical samples by using reverse transcriptase-polymerase chain reaction. J Clin Microbiol 30: 545551.

    • Search Google Scholar
    • Export Citation
  • 21.

    Medellín RA, Equihua M, Amin M, 2000. Bat diversity and abundance as indicators of disturbance in neotropical rainforest. Conserv Biol 14: 16661675.

    • Search Google Scholar
    • Export Citation
  • 22.

    Vargas J, Medellín RA, Escalona-Segura G, Interián-Sosa L, 2009. Vegetation complexity and bat-plant dispersal in Calakmul, Mexico. J Nat Hist 43: 219243.

    • Search Google Scholar
    • Export Citation
  • 23.

    Estrada A, Coates-Estrada R, Meritt D, 1993. Bat species richness and abundance in a tropical rain forest fragments and in agricultural habitats at Los Tuxtlas, Mexico. Ecography (Cop.) 16: 309318.

    • Search Google Scholar
    • Export Citation
  • 24.

    R Core Team, 2013. R: A Language and Environment for Statistical Computing. Vienna, Austria: R Foundation for Statistical Computing.

  • 25.

    Gómez Dantes H, 2007. Elementos económicos y políticos que impactan en el control del dengue en México. Salud Publica Mex 49: 117119.

    • Search Google Scholar
    • Export Citation

Author Notes

* Address correspondence to Jesús Sotomayor-Bonilla, Universidad 3000, Colonia Ciudad Universitaria, México, Distrito Federal, Mexico 03310. E-mail: chuchomayor16@gmail.com

Financial support: This research was supported by funding from the US Agency for International Development (USAID) Emerging Pandemic Threats PREDICT Project. The Medical Immunology Research Unit, Health Research Coordination Office, Mexican Institute for Social Security supported the RNA extraction. The Genetic Conservation Laboratory, Biology School, University of Costa Rica supported diagnostic tests. The Conservation Medicine Laboratory of the National Polytechnic Institute supported confirmation tests. J.S.-B. was supported by CONACYT Grant 303956.

Authors' addresses: Jesús Sotomayor-Bonilla, Oscar Rico-Chávez, Rafael Ojeda-Flores, and Gerardo Suzán, Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México—Etología, Fauna Silvestre y Animales de Laboratorio, México, DF, Mexico, E-mails: chuchomayor16@gmail.com, orichvet@gmail.com, ojeda_rafael@yahoo.com.mx, and gerardosuz@gmail.com. Andrea Chaves, Escuela de Biología, Universidad de Costa Rica Laboratorio de Genética de la Conservación, San José, Costa Rica, E-mail: andreachaves.biol@gmail.com. Melinda K. Rostal and Peter Daszak, EcoHealth Alliance, New York, NY, E-mails: rostal@ecohealthalliance.org and daszak@ecohealthalliance.org. Mónica Salas-Rojas and Álvaro Aguilar-Setien, Coordinación de Investigación en Salud, Instituto Mexicano del Seguro Social, Unidad de Investigación Médica en Inmunología, México, DF, Mexico, E-mails: mony_salas@yahoo.com.mx and varoaguila@prodigy.net.mx. Sergio Ibáñez-Bernal, Instituto de Ecología AC, Red Ambiente y Sustentabilidad, Xalapa, Mexico, E-mail: sibanber@gmail.com. Arturo Barbachano-Guerrero, Gustavo Gutiérrez-Espeleta, and J. Leopoldo Aguilar-Faisal, Escuela Superior de Medicina, Instituto Politécnico Nacional, Laboratorio de Medicina de Conservación, México, DF, Mexico, E-mails: abarbachanog@gmail.com, gutierrezespeleta@gmail.com, and leopoldoaguilar@hotmail.com. A. Alonso Aguirre, Smithsonian–Mason School of Conservation, Front Royal, VA, E-mail: aaguirr3@gmu.edu.

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