Fil: Fernández, María Soledad. Instituto Nacional de Medicina Tropical (INMeT). Puerto Iguazú. Misiones; ArgentinaFil: Santini, María Soledad. Centro Nacional de Diagnóstico e Investigaciones Endemo-epidemicas (CeNDIE). Administración Nacional de Laboratorios e Institutos de Salud (ANLIS); ArgentinaFil: Díaz, Julia Inés. Centro de Estudios en Parasitología y Vectores (CEPAVE). Facultad de Ciencias Naturales y Museo. Universidad Nacional de La Plata; ArgentinaFil: Villarquide, Lucrecia. Laboratorio de Control de Vectores Entomológicos de Importancia Sanitaria (LaCVEIS) Fundación H. A.Barceló; ArgentinaFil: Lestani, Eduardo. Instituto Nacional de Medicina Tropical (INMeT). Puerto Iguazú. Misiones; ArgentinaFil: Salomón, Oscar Daniel. Instituto Nacional de Medicina Tropical (INMeT). Puerto Iguazú. Misiones; ArgentinaFil: Achinelly, María Fernanda. Centro de Estudios en Parasitología y Vectores (CEPAVE). Facultad de Ciencias Naturales y Museo. Universidad Nacional de La Plata; Argentin

Achinelly, María Fernanda

Díaz, Julia Inés

Fernández, María Soledad

Lestani, Eduardo

Salomón, Oscar Daniel

Santini, María Soledad

Villarquide, Lucrecia

Fil: Fernández, María Soledad. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Medicina Tropical; Argentina.Fil: Santini, María Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina.Fil: Diaz, Julia I. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina.Fil: Villarquide, L. Fundación H. A. Barceló. Laboratorio de Control de Vectores Entomológicos de Importancia Sanitaria (LaCVEIS);  Argentina.Fil: Lestani, Eduardo. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Medicina Tropical; Argentina.Fil: Salomón, Oscar Daniel. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Medicina Tropical; Argentina.Fil: Achinelly, María Fernanda. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina.Pintomyia fischeri adults collected in different eco-epidemiological studies in the northeastern of Argentina
were found parasitized by juvenile nematodes (Tylenchida) isolated from the body cavity. The percentage of
infected females and males was 3.8% and 2.9% respectively. Part of the life cycle of sand flies and tylenchid
nematodes take place in humid and dark sites, where infection of immature stage of Phlebotominae insects is
possible. Biology of this parasite could help to determine the breeding sites of sand flies. This study constituted the first report of tylenchid nematodes infecting sand flies at field conditions in South-America

Diaz, Julia I

Villarquide, L

Sistema de Gestión del Conocimiento ANLIS MALBRÁN

Parasitism by Tylenchid Nematodes in Natural Populations of Pintomyia fischeri (Diptera: Psychodidae: Phlebotominae) in Argentina

Naturalis

SM Tropical Medicine JournalGr   upSMHow to cite this article Fernández MS, Santini MS, Diaz JI, Villarquide L, Lestani E and Achinelly M et al. Parasitism by Tylenchid Nematodes in Natural Populations of Pintomyia fischeri (Diptera: Psychodidae: Phlebotominae) in Argentina. SM Trop Med J. 2015;1(1):1001.OPEN ACCESSPhlebotominae are insects of public health importance because their role in the transmission of the etiological agents of several diseases, being the different leishmaniases the best known around the world. There are 28 species of Phlebotominae recorded in Argentina [1], some of them involved in the transmission of the etiological agent of Visceral Leishmaniasis (VL) and Cutaneous Leishmaniasis (CL) [2]. Pintomyia fischeri (Pinto, 1926) constitutes the third more abundant species in a deforestation area in northeast Argentina [3] where CL due to Leishmania braziliensis Vianna, 1911, is endemic [4,5]. There are not records of Pi. fischeri as Leishmania (Viannia) spp. vector in Argentina [1], although adults infected by Leishmania spp. have been observed in an endemic area of ACL in Brazil, suggesting this sand fly species as a probable secondary vector [6]. Taking into account the public health importance of sand flies, any contribution regarding a possible agent for its biological control is relevant.Tylenchid nematodes were reported from the body cavity of sand flies [7]. The infective stage (gravid females) penetrates through the body wall of first stage larvae of phlebotomine insects. The eggs develop to third and fourth juvenile stage, and they are accumulated in the infected sand fly adults. Nematodes leave the insect host via the anus or reproductive pore and mature to adult in the environment where mated females reinvade the insect to restart the cycle. Heavy infections greatly reduced egg production and in several cases, completely sterilized female flies. So, these nematodes could be a natural bioregulator agent of sand fly populations [8]. In this work parasitism by tylenchid nematodes is reported in natural populations of the sand fly Pi. fischeri, in the northeastern Argentina. The study was carry out in a rural zone of Puerto Iguazú (25º 39` S, 54º 33 W), Misiones province, at the northeastern border of Argentina. The area known as “Dos mil hectáreas” , which covers an area of 2000 hectares, was originally a native forest, but after its partial deforestation, many farms have been established since 2003, with a peak of human intervention and settlement during 2004 [4]. A mosaic composed by primary and secondary forest, pigsties, chicken houses, crops and houses (farms) arrange the landscape of the area which is surrounded by natural protected forest reservations and the Paraná River.Phlebotominae were collected in the framework of different eco-epidemiological studies from winter 2006 to summer 2010 using CDC mini light traps (408 traps-night) operated from 5:00 PM to 09:00 AM and placed at 1.5 m above the ground [9]. Sand flies were preserved in 1% lactophenol and identified according to Galati (2003).Short CommunicationParasitism by Tylenchid Nematodes in Natural Populations of Pintomyia fischeri (Diptera: Psychodidae: Phlebotominae) in ArgentinaFernández MS1,2, Santini MS2,3, Diaz JI2,4, Villarquide L5, Lestani E1, Salomón OD1,2 and Achinelly M2,4*1Instituto Nacional de Medicina Tropical (INMeT), Puerto Iguazú, Misiones, Argentina 2Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina3Centro Nacional de Diagnóstico e Investigaciones Endemo-epidemicas (CeNDIE), Administración Nacional de    Laboratorios e Institutos de Salud (ANLIS), Argentina4Centro de Estudios Parasitológicos y de Vectores, CCT La Plata (CONICET-UNLP), Argentina 5Laboratorio de Control de Vectores Entomológicos de Importancia Sanitaria (LaCVEIS) Fundación H. A. Barceló, ArgentinaArticle InformationReceived date: Oct 22, 2015 Accepted date: Oct 26, 2015 Published date: Oct 30, 2015*Corresponding authorAchinelly María Fernanda, CEPAVE-CCT-La Plata-CONICET-UNLP, Argentina; Email: fachinelly@cepave.edu.ar Distributed under Creative Commons CC-BY 4.0Keywords Sand Flies; Nematoda; Tylenchida Larvae; Field ConditionsAbstractPintomyia fischeri adults collected in different eco-epidemiological studies in the northeastern of Argentina were found parasitized by juvenile nematodes (Tylenchida) isolated from the body cavity. The percentage of infected females and males was 3.8% and 2.9% respectively. Part of the life cycle of sand flies and tylenchid nematodes take place in humid and dark sites, where infection of immature stage of Phlebotominae insects is possible. Biology of this parasite could help to determine the breeding sites of sand flies. This study constituted the first report of tylenchid nematodes infecting sand flies at field conditions in South-America.Citation: Fernández MS, Santini MS, Diaz JI, Villarquide L, Lestani E and Achinelly M et al. Parasitism by Tylenchid Nematodes in Natural Populations of Pintomyia fischeri (Diptera: Psychodidae: Phlebotominae) in Argentina. SM Trop Med J. 2015;1(1):1001. Page 2/3Gr   upSM Copyright  Achinelly MNematodes recovered from the body cavity of sand flies were identified using a light microscope, following specific bibliography [11,12]. Photographs were taken using a Q-Imaging Go-3 camera. The length and width were determined; measurements were given in micrometers, with the mean followed by the range in brackets. A total of 898 Pi. fischeri adults were captured in 97 trap-night (557 females and 341 males). Thirty-one Pi. fischeri specimens were found parasitized by filiform juvenile nematodes in the haemocele representing the 3.5% of the total Pi. fischeri capture (14 light traps). The percentage of infected females and males was 3.8% and 2.9% respectively. Nematodes presented buccal stylet and pharyngeal glands characteristic of Tylenchida larvae (Figures 1 A-D). The mean length (n=98) was 233 μm (160-263), and the width 17.6 μm (10-23). Unfortunately due to the nature of the sand flies diaphanization methodology -lactophenol-, which prevented the maturation of juvenile nematodes to female adult stage, taxonomic identification was not possible. Parasitized individuals were collected in peridomestic capture sites (henhouse and pigsties), except one from the edge of forest. They were captured mainly in summer (n=23), but with some records in spring (n=5) and fall (n=3). Poinar et al. (1993) [8] observed that the elimination of tylenchid nematode juveniles and parasite infection could be take place in humid and dark sites, similar than those suggested for the oviposition and larval growing of Phlebotominae as tree buttresses, and rodent burrows with organic debris like leaf litter, dung and crevices [7]. Pintomya ficheri was found in Argentina in a secondary forest and peridomestic environments [3,4]. In the same way, Rangel and Lainson (2009) [13], suggested that this species in Brazil is adapting to the domiciliary environment, with anthropophilic behavior, but also feeding on dogs and birds. Although the breeding sites of the immature stages of Pi. fischeri in Argentina has not been found yet, it could take place under humidity and darkness soil conditions at the sampling sites considered in our study. Knowledge of the biology and dynamics of this nematode at field may be useful for determining the breeding sites of the immature stages of these insects, which is necessary to develop control strategies. Pathogenicity and deleterious effects on the reproductive success of Pi. fischeri (and potentially other sand flies species), could contribute to look for new ways for vector bioregulation. Tylenchid nematodes were reported from different species of Phlebotominae sand flies from Africa, the Middle East and Central America [7,14-16]. Individuals of Lutzomyia longipalpis (Lutz & Neiva 1912) were observed infected in a laboratory colony where progenitors were originally collected in Cundinamarca, Colombia [8].This report constituted as our knowledge the first report of tylenchid nematodes parasitizing natural populations of Phlebotominae in South-America. Acknowledgement We are very grateful to the inhabitants of “Dos mil hectáreas” for providing access to the farms, Fundación Mundo Sano and Dr. Cecilia Carballo for her assistance in the early stages of this study.References 1. Salomón OD, Quintana MG. Leishmaniasis y ambiente: uso de sensores remotos y sistemas de información geográfica en Temas de Zoonosis V. Basualdo J, Cacchione R, Durlach R, Martino P, Seijo A, editors. In: Sociedad Argentina de Zoonosis. 2011. 2. Salomón OD, Quintana MG, Rosa JR. Eco-epidemiologia de leishmaniasis cutánea en Argentina. Soc Iberoam Inf Científica (SIICSALUD). 2008; 16: 514-520. 3. Fernández MS, Lestani EA, Cavia R, Salomón OD. Phlebotominae fauna in a recent deforested area with American Tegumentary Leishmaniasis transmission (Puerto Iguazú, Misiones, Argentina): Seasonal distribution in domestic and peridomestic environments. Acta Trop. 2012; 122: 16-23.4. Salomón OD, Acardi SA, Liotta DJ, Fernández MS, Lestani E, et al. Epidemiological aspects of cutaneous leishmaniasis in the Iguazú falls area of Argentina. Acta Trop. 2009; 109: 5-11. 5. Salomón OD, Sosa Estani S. Brote epidémico de Leishmaniasis tegumentaria en Puerto Esperanza, 1998. Prov Misiones. Medicina (Buenos Aires). 2001; 61: 385-390. 6. Pita-Pereira D de, Souza GD, Araújo Pereira T de, Zwetsch A, Britto C, Rangel EF. Lutzomyia (Pintomyia) fischeri (Diptera: Psychodidae: Phlebotominae), a probable vector of American cutaneous leishmaniasis: detection of natural infection by Leishmania (Viannia) {DNA} in specimens from the municipality of Porto Alegre ({RS}), Brazil, using multiplex PCR assay. Acta Trop. 2011; 120: 273-275.7. Dinesh DS, Kumar V, Das P. Infestation of Nematodes in Phlebotomus Argentipes Annandale and Brunetti (Diptera: Psycodidae), Bihar, India. Glob J Med Res. 2013; 13. 8. Poinar GO, Ferro C, Morales A, Tesh RB. Anandranema phlebotophaga n. gen., n. sp.(Allantonematidae: Tylenchida), a new nematode parasite of phlebotomine sand flies (Psychodidae: Diptera) with notes on experimental infections of these insects with parasitic rhabditoids. Fundam Appl Nematol. 1993; 16: 11-16. 9. Sudia WD, Chamberlain RW. Battery operated light trap, an improved model. Mosq News. 1962; 22: 126-129. 10. Galati EAB. Morfologia, terminologia de adultos e identificao dos taxons da América. Rangel EF, Lainson R, editors. In: Flebotomíneos do Brasil. Fiocruz, 2003. 11. Poinar GO. C.I.H. Key to the Groups and Genera of Nematode Parasites of Figure 1: A) and B) Infected Pintomyia fischeri female, posterior end. C) Nematode larvae released from the host. D) Detail of nematode larva, anterior end, showing buccal stylet (arrow).Citation: Fernández MS, Santini MS, Diaz JI, Villarquide L, Lestani E and Achinelly M et al. Parasitism by Tylenchid Nematodes in Natural Populations of Pintomyia fischeri (Diptera: Psychodidae: Phlebotominae) in Argentina. SM Trop Med J. 2015;1(1):1001. Page 3/3Gr   upSM Copyright  Achinelly MInvertebrates. Farnham Royal, Bucks: Common w Agric Bur. 1977; 43.12. Siddiqi MR. Tylenchida: parasites of plants and insects. CABI. 2000.13. Rangel EF, Lainson R. Proven and putative vectors of American cutaneous leishmaniasis in Brazil: aspects of their biology and vectorial competence. Mem Inst Oswaldo Cruz. 2009; 104: 937-954. 14. McConnell E, Correa M. Trypanosomes and other microorganisms from Panamanian Phlebotomus sandflies. J Parasitol. 1964; 523-528. 15. Karakus M, Arserim SK, Töz SÖ, Özbel Y. Detection of Entomopathogen Nematode [EPN-Sand Flies (Phlebotomus tobbi)] Caught in the Wild in Aydin, Kusadasi Town and its Assessment as a Biological Control Agent. Türkiye Parazitol Derg. 2013; 37: 36-39. 16. Secundino NFC, Araújo MSS, Oliveira GHB, Massara CL, Carvalho OS, Lanfredi RM, et al. Preliminary description of a new entomoparasitic nematode infecting Lutzomyia longipalpis sand fly, the vector of visceral leishmaniasis in the New World. J Invertebr Pathol. 2002; 80: 35-40.

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Parasitism by Tylenchid Nematodes in Natural Populations of Pintomyia fischeri (Diptera: Psychodidae: Phlebotominae) in Argentina

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