Leishmania DNA detection and species characterization within phlebotomines (Diptera: Psychodidae) from a peridomicile-forest gradient in an Amazonian/Guianan bordering area

In the border region between Brazil and French Guiana, American cutaneous leishmaniasis is a worrisome public health issue, and entomological studies are required there to better identify classical and putative emerging transmission patterns. The present study aimed to detect and characterize Leishmania DNA in the phlebotomine population of Oiapoque (Amapá State, Brazil). Phlebotomines were captured in anthropized and wild environments in the outskirts of Oiapoque municipality, using CDC light traps installed in vertical (ground/canopy level) and horizontal (peridomicile/extradomicile/forest-edge/forest) strata. Captured specimens were identified according to their morphology. Females were processed for Leishmania DNA detection and characterization using a multiplex polymerase chain reaction targeting kinetoplast DNA (kDNA) and the phlebotomine cacophony gene. The kDNA positive samples were characterized by cloning and sequencing the Leishmania 234 bp-hsp70 gene. Among the 3957 phlebotomine specimens captured, 26 pooled female samples were positive for Leishmania (Viannia) spp. DNA. Sequencing analysis allowed species-specific identification of L. (V.) braziliensis DNA in Trichophoromyia ininii, Bichromomyia flaviscutellata, Nyssomyia umbratilis, and Evandromyia infraspinosa, and L. (V.) guyanensis DNA in Ny. umbratilis. A pooled sample of Ny. umbratilis was positive for both L. (V.) braziliensis and L. (V.) guyanensis DNA. The present study provided additional information regarding ACL ecology in Oiapoque, highlighting the presence of L. (V.) braziliensis DNA in different phlebotomine species. The epidemiological implications of these findings and the determinant incrimination of L. (V.) braziliensis as proven vectors in that region must be clarified. In this regard, studies on Leishmania spp. infection and suggestive anthropophilic behavior of associated phlebotomines need to be prioritized in entomological surveillance.

[1]  E. F. Rangel,et al.  Binational burden of American cutaneous leishmaniasis in Oiapoque, Amapá State, Brazil, bordering French Guiana. , 2019, Revista da Sociedade Brasileira de Medicina Tropical.

[2]  E. F. Rangel,et al.  Ecological aspects of Phlebotomines (Diptera: Psychodidae) and the transmission of American cutaneous leishmaniasis agents in an Amazonian/ Guianan bordering area , 2018, Parasites & Vectors.

[3]  M. M. Vale,et al.  Entomological Surveys of Lutzomyia flaviscutellata and Other Vectors of Cutaneous Leishmaniasis in Municipalities with Records of Leishmania amazonensis within the Bragança Region of Pará State, Brazil , 2018, Journal of Vector Ecology.

[4]  Anne-Laure Bañuls,et al.  iDNA screening: Disease vectors as vertebrate samplers , 2017, Molecular ecology.

[5]  P. Lawyer,et al.  An illustrated guide for characters and terminology used in descriptions of Phlebotominae (Diptera, Psychodidae) , 2017, Parasite.

[6]  Mathieu Nacher,et al.  Cutaneous leishmaniasis in French Guiana: revising epidemiology with PCR-RFLP , 2017, Tropical Medicine and Health.

[7]  Daniela de Pita-Pereira,et al.  First description of Leishmania (Viannia) infection in Evandromyia saulensis, Pressatia sp. and Trichophoromyia auraensis (Psychodidae: Phlebotominae) in a transmission area of cutaneous leishmaniasis in Acre state, Amazon Basin, Brazil , 2017, Memorias do Instituto Oswaldo Cruz.

[8]  J. Shaw,et al.  High Resolution Melting Analysis Targeting hsp70 as a Fast and Efficient Method for the Discrimination of Leishmania Species , 2016, PLoS neglected tropical diseases.

[9]  S. Luz,et al.  Anthropic effects on sand fly (Diptera: Psychodidae) abundance and diversity in an Amazonian rural settlement, Brazil. , 2014, Acta tropica.

[10]  L. Nakazato,et al.  Phlebotomine fauna, natural infection rate and feeding habits of Lutzomyia cruzi in Jaciara, state of Mato Grosso, Brazil , 2014, Memorias do Instituto Oswaldo Cruz.

[11]  A. L. R. Roque,et al.  Wild and synanthropic reservoirs of Leishmania species in the Americas , 2014, International journal for parasitology. Parasites and wildlife.

[12]  P. Volf,et al.  Speculations on biting midges and other bloodsucking arthropods as alternative vectors of Leishmania , 2014, Parasites & Vectors.

[13]  E. F. Rangel,et al.  Leishmaniasis transmission in an ecotourism area: potential vectors in Ilha Grande, Rio de Janeiro State, Brazil , 2013, Parasites & Vectors.

[14]  D. Mans,et al.  Studies on the sand fly fauna (Diptera: Psychodidae) in high-transmission areas of cutaneous leishmaniasis in the Republic of Suriname , 2013, Parasites & Vectors.

[15]  O. Faccenda,et al.  Fauna of phlebotomine sand flies (Diptera, Psychodidae) in areas with endemic American cutaneous leishmaniasis in the State of Mato Grosso do Sul, Brazil , 2013 .

[16]  P. Ready.  Biology of phlebotomine sand flies as vectors of disease agents. , 2013, Annual review of entomology.

[17]  P. Volf,et al.  Development of Leishmania Parasites in Culicoides nubeculosus (Diptera: Ceratopogonidae) and Implications for Screening Vector Competence , 2012, Journal of medical entomology.

[18]  A. Volpini,et al.  Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species. , 2012, Memorias do Instituto Oswaldo Cruz.

[19]  E. F. Rangel,et al.  Phlebotomine Fauna (Diptera: Psychodidae) and Putative Vectors of Leishmaniases in Impacted Area by Hydroelectric Plant, State of Tocantins, Brazil , 2011, PloS one.

[20]  A. L. R. Roque,et al.  Thrichomys laurentius (Rodentia; Echimyidae) as a Putative Reservoir of Leishmania infantum and L. braziliensis: Patterns of Experimental Infection , 2010, PLoS neglected tropical diseases.

[21]  V. L. Nunes,et al.  Epidemiological study on leishmaniasis in an area of environmental tourism and ecotourism, State of Mato Grosso do Sul, 2006-2007. , 2009, Revista da Sociedade Brasileira de Medicina Tropical.

[22]  J. Shaw,et al.  A Phylogenetic Lineage of Closely Related Trypanosomes (Trypanosomatidae, Kinetoplastida) of Anurans and Sand Flies (Psychodidae, Diptera) Sharing the Same Ecotopes in Brazilian Amazonia 1 , 2008, The Journal of eukaryotic microbiology.

[23]  C. Ravel,et al.  Phlebotomine sand flies (Diptera: Psychodidae) associated with changing patterns in the transmission of the human cutaneous leishmaniasis in French Guiana. , 2007, Memorias do Instituto Oswaldo Cruz.

[24]  Daniela de Pita-Pereira,et al.  Identification of naturally infected Lutzomyia intermedia and Lutzomyia migonei with Leishmania (Viannia) braziliensis in Rio de Janeiro (Brazil) revealed by a PCR multiplex non-isotopic hybridisation assay. , 2005, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[25]  A. Franco,et al.  Feeding preference of the sand flies Lutzomyia umbratilis and L. spathotrichia (diptera: Psychodidae, Phlebotominae) in an urban forest patch in the city of Manaus, Amazonas, Brazil. , 2004, Memorias do Instituto Oswaldo Cruz.

[26]  E. Cupolillo,et al.  Genetic Polymorphism and Molecular Epidemiology of Leishmania (Viannia) braziliensis from Different Hosts and Geographic Areas in Brazil , 2003, Journal of Clinical Microbiology.

[27]  M. Brito,et al.  Wild and synanthropic hosts of Leishmania (Viannia) braziliensis in the endemic cutaneous leishmaniasis locality of Amaraji, Pernambuco State, Brazil. , 2003, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[28]  C. Kyriacou,et al.  Molecular evolution of the cacophony IVS6 region in sandflies , 2002, Insect molecular biology.

[29]  X. Huang,et al.  CAP3: A DNA sequence assembly program. , 1999, Genome research.

[30]  D. Haburchak,et al.  Topley and Wilson's Microbiology and Microbial Infections , 1999 .

[31]  P. Green,et al.  Base-calling of automated sequencer traces using phred. I. Accuracy assessment. , 1998, Genome research.

[32]  E. F. Rangel,et al.  Variation between geographical populations of Lutzomyia (Nyssomyia) whitmani (Antunes & Coutinho, 1939) sensu lato (Diptera:Psychodidae:Phlebotominae) in Brazil. , 1996, Memorias do Instituto Oswaldo Cruz.

[33]  W. Degrave,et al.  An oligonucleotide probe derived from kDNA minirepeats is specific for Leishmania (Viannia). , 1996, Memorias do Instituto Oswaldo Cruz.

[34]  J. Shaw,et al.  The dermal leishmaniases of Brazil, with special reference to the eco-epidemiology of the disease in Amazonia. , 1994, Memorias do Instituto Oswaldo Cruz.

[35]  H. Guerra,et al.  Natural Leishmania infection of Lutzomyia spp. in Peru. , 1994, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[36]  U. Lopes,et al.  The identity of Leishmania isolated from sand flies and vertebrate hosts in a major focus of cutaneous leishmaniasis in Baturite, northeastern Brazil. , 1994, The American journal of tropical medicine and hygiene.

[37]  J. Dedet.  Cutaneous leishmaniasis in French Guiana: a review. , 1990, The American journal of tropical medicine and hygiene.

[38]  R. Killick-Kendrick.  Phlebotomine vectors of the leishmaniases: a review , 1990, Medical and veterinary entomology.

[39]  J. Shaw,et al.  The ecology of lutzomyia umbratilis Ward & Fraiha (Diptera: Psychodidae), the major vector to man of Leishmania braziliensis guyanensis in north-eastern Amazonian brazil , 1986 .

[40]  J. Arias,et al.  Hosts of sandfly vectors of Leishmania braziliensis guyanensis in the central Amazon of Brazil. , 1982, The American journal of tropical medicine and hygiene.

[41]  F. Sanger.  The Croonian Lecture, 1975 Nucleotide sequences in DNA , 1975, Proceedings of the Royal Society of London. Series B. Biological Sciences.

[42]  Anabela S Rodrigues,et al.  Sand Fly Vectors of Leishmania in the Americas - A Mini Review , 2014 .

[43]  J. L. P. Moraes,et al.  [Occurrence of sand flies (Diptera, Psychodidae) in leishmaniasis foci in an ecotourism area around the Lençóis Maranhenses National Park, Brazil]. , 2010, Cadernos de saude publica.

[44]  P. Volf,et al.  Leishmania in Sand Flies: Comparison of Quantitative Polymerase Chain Reaction with Other Techniques to Determine the Intensity of Infection , 2008, Journal of medical entomology.

[45]  B. Rotureau.  Ecology of the leishmania species in the Guianan ecoregion complex. , 2006, The American journal of tropical medicine and hygiene.

[46]  O Hammer-Muntz,et al.  PAST: paleontological statistics software package for education and data analysis version 2.09 , 2001 .

[47]  W. Degrave,et al.  Natural infection of a domestic cat (Felis domesticus) with Leishmania (Viannia) in the metropolitan region of Belo Horizonte, State of Minas Gerais, Brazil. , 1996, Memorias do Instituto Oswaldo Cruz.

[48]  J. Dedet,et al.  [Evaluation of a cutaneous leishmaniasis control program in a forest village of French Guyana]. , 1986, Bulletin of the World Health Organization.

[49]  J. Dedet,et al.  Evaluation d'un programme de lutte contre la leishmaniose cutanée dans un village forestier de Guyane française , 1986 .

[50]  J. Shaw,et al.  Leishmaniasis in Brazil: XX. Prevalence of "enzootic rodent leishmaniasis" (Leishmania mexicana amazonensis), and apparent absence of "pian bois" (Le. braziliensis guyanensis), in plantations of introduced tree species and in other non-climax forests in eastern Amazônia. , 1983, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[51]  J. Shaw,et al.  Leishmaniasis in Brazil: XVI. Isolation and identification of Leishmania species from sandflies, wild mammals and man in north Para State, with particular reference to L. braziliensis guyanensis causative agent of "pian-bois". , 1981, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[52]  J. Shaw,et al.  Leishmaniasis in Brazil: XIII. Isolation of Leishmania from armadillos (Dasypus novemcinctus), and observations on the epidemiology of cutaneous leishmaniasis in north Pará State. , 1979, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[53]  J. Shaw,et al.  Leishmaniasis in Brazil. VI. Observations on the seasonal variations of Lutzomyia flaviscutellata in different types of forest and its relationship to enzootic rodent leishmaniasis (Leishmania mexicana amazonensis). , 1972, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[54]  J. Shaw,et al.  Leishmaniasis in Brazil: II. Observations on enzootic rodent leishmaniasis in the Lower Amazon Region--the feeding habits of the vector, Lutzomyia flaviscutellata in reference to man, rodents and other animals. , 1968, Transactions of the Royal Society of Tropical Medicine and Hygiene.