Focus Of Cutaneous Leishmaniasis Research Articles

PCR detection and sequencing of parasite ITS-rDNA gene from reservoirs host of zoonotic cutaneous leishmaniasis in central Iran

Leishmania major is the causative agent of zoonotic cutaneous leishmaniasis (ZCL) in which gerbils are the reservoir host. ZCL is of great public health importance in Iran. In the current investigation, nested polymerase chain reaction (PCR) protocols were used to amplify a region of the ribosomal RNA amplicon of Leishmania (ITS1-5.8S rRNA gene). The PCR assays detected L. major in three rodent species: Rhombomis opimus, Meriones lybicus and, for first time, Meriones persicus. L. major parasite was found in Natanz, Isfahan Province in the center of Iran in a focus of rural zoonotic cutaneous leishmaniasis. Four L. major infections were detected in R. opimus species, three in M. Lybicus, and two in M. persicus. All nine rodent infections of L. major were found to be the same haplotype based on the PCR detection and sequencing of parasite ITS-ribosomal DNA gene. In addition, also for the first time, the nested PCR assays detected Leishmania tropica only in one M. persicus. Allied to studies in country, the new findings mean that past conclusions about the reservoir of L. major in Iran must be treated with caution. Finding two Leishmania species in different rodent species as reservoir in Iran, therefore, careful molecular eco-epidemiological investigations will be an essential part of modeling the roles of different gerbil species in maintaining and spreading ZCL foci.

Primer informe de Lutzomyia yuilli Young & Porter, 1972 y Lutzomyia triramula (Fairchild & Hertig 1952) (Diptera: Psychodidae) en el departamento de Caldas, Colombia

Durante estudios de foco en varios municipios del departamento de Caldas, Colombia, donde se han presentado casos de leishmaniosis cutánea, se recolectaron nueve especies de Lutzomyia: Lutzomyia gomezi (Nitzulescu 1931), Lu. ayrozai (Barreto & Coutinho 1940), Lu. panamensis (Shannon 1926), Lu. bifoliata Osorno, Morales, Osorno & Hoyos, 1970, Lu. trapidoi (Farchild & Hertig 1952), Lu. yuilli Young & Porter, 1952, Lu. triramula (Fairchild & Hertig 1952), Lu. atroclavata (Knab 1913) y Lu. trinidadensis (Newstead 1922). Dos de estos taxones constituyen nuevos registros para el departamento, Lu. yuilli del subgénero Nyssomyia, y Lu. triramula del subgénero Trichopygomyia. Los individuos de Lu. yuilli se obtuvieron mediante el uso de trampas de luz tipo Shannon y CDC en el municipio de Norcasia, mientras que los de Lu. triramula se recolectaron usando trampas de luz y adhesivas en los municipios de Norcasia y Victoria. Se discute la importancia de este hallazgo teniendo en cuenta la limitada distribución de Lu. triramula en el territorio colombiano y la capacidad vectorial de Lu. yuilli.

Nested PCRs and sequencing of nuclear ITS‐rDNA fragments detect three Leishmania species of gerbils in sandflies from Iranian foci of zoonotic cutaneous leishmaniasis

To identify and understand the natural transmission cycles of Leishmania in Iranian sandflies. Nested PCR protocols were developed to amplify two regions of the ribosomal RNA amplicon of Leishmania (ITS1-5.8S rRNA gene, and a microsatellite DNA region of ITS2), which were species-specific by DNA sequence or fragment size. The PCR assays detected in Iranian sandflies not only Leishmania major but also for the first time L. turanica and L. gerbilli sensu lato, two other parasites of the great gerbil. All three parasites were found in the northeast and centre of Iran, in two foci of rural Zoonotic Cutaneous Leishmaniasis (ZCL) caused by L. major. Fifty infections were detected in common sandfly species: at least six geographically differentiated haplotypes of L. major in four to five sandfly species; one strain of L. gerbilli s.l. in five to six sandfly species and one strain of L. turanica in one sandfly species. Past conclusions about the transmission cycles of L. major in Iran should be treated with caution. Careful molecular eco-epidemiological investigations are essential for modelling the roles of different sandfly species in maintaining and spreading ZCL foci. Even if non-pathogenic to humans, frequent inoculations of L. turanica by sandflies might alter the efficacy of vaccines against L. major. Phlebotomus papatasi is probably the key vector in many ZCL foci because of its abundance and high infection rates with both L. major and L. turanica.

Entomological and epidemiological study of a new focus of cutaneous leishmaniasis in Morocco

Cutaneous leishmaniasis (CL) due to Leishmania tropica Wright has become an increasingly important problem in Al-Haouz province, Morocco. The objective of this paper was to undertake a retrospective analysis of CL between 2000 and 2006 and to study the sandfly fauna for the first time in this focus. The consultation of bulletins, registers and monthly and annual reports published by local and national medical services permitted a global synthesis of CL in Al-Haouz between 2000 and 2006. Sandflies were collected using sticky-paper traps in five selected stations in two communes, Ourika and Sti Fadma, the most affected by CL in Al-Haouz. Overall, 2,648 sandflies, belonging to five species of Phlebotomus and three of Sergentomyia, were collected. Phlebotomus sergenti Parrot (14.5%) was found in all studied stations where it was associated with domestic and peridomestic habitats. Based on this first report on ecology of sandflies and epidemiology of CL, Al-Haouz seems to be a typical focus of L. tropica and P. sergenti like the other Moroccan foci. Two communes, Sti Fadma and Ourika, constituted an important CL micro-focus in this region. Considering their economical importance (trade and tourism), they contribute probably to the dissemination of Leishmania parasite in bordering regions.

Urban distribution of Phlebotominae in a cutaneous leishmaniasis focus, Argentina

Urbanization and vector domestication are currently proposed as factors that contributed to the recent increase of American cutaneous leishmaniasis (ACL). Is likely also urban transmission? Oran is the main city in the Argentinean hyper-endemic area of ACL, and human cases in urban residences are usually reported. In order to assess the spatial distribution of risk, phlebotomine traps were located in different environments of Oran. A total of 7,787 sand flies were captured: Lutzomyia neivai (98.1%), Lutzomyia migonei (1.2%), Lutzomyia cortelezzii (0.7%), and one Lutzomyia shannoni. During the season of transmission (April-May) a single sand fly was obtained in one out of five urban sites, while a trap in a peri-urban pigsty captured up to 2,985 Lu. neivai/night. Captures performed in the other season of vector activity (September-October) revealed that small-scale changes in the pigsty environment resulted in noticeable changes in the abundance of Lu. neivai. In addition, in a new neighbourhood, on the fringe of the city, 1,073 Lu. neivai/site were captured in the forested edge but one in the yard of the houses. Therefore, in this urban ACL focus the human-vector effective contact risk is still associated with peri-urban vegetation and ecotone modifications despite the urban residence of the cases.

Rodent infection withLeishmaniain a new focus of human cutaneous leishmaniasis, in northern Iran

The incidence of zoonotic cutaneous leishmaniasis (ZCL), which is endemic in several parts of Iran, has recently increased in the rural areas of the northern district of Damghan, in Semnan province. Between 2001 and 2004, as part of an investigation of this worrying trend, wild rodents in this region were caught and checked, by the microscopical examination of liver and spleen smears, for leishmanial infection, to see which species were acting as 'reservoir' hosts. Overall 298 Nesokia indica, 29 Meriones libycus, 10 Mus musculus and two Microtus socialis were caught. Most of the N. indica (61%) and Me. libycus (52%) but none of the other rodents were found smear-positive for leishmanial amastigotes. When PCR was used to test scrapings from 50 of the smears (of which 25 had been found to harbour amastigotes, by microscopy), 29 (58%) of the smears were found positive for the kinetoplast DNA of Leishmania major; none was found positive for any other Leishmania species. Nesokia indica and Me. libycus are therefore incriminated as the main 'reservoir' hosts of L. major in Damghan.

Some Ecological Aspects of Phlebotominae Sand Flies (Diptera: Psychodidae) in an Endemic Focus of Anthroponotic Cutaneous Leishmaniasis of Iran

Some Ecological Aspects of Phlebotominae Sand Flies (Diptera: Psychodidae) in an Endemic Focus of Anthroponotic Cutaneous Leishmaniasis of Iran

New Clinicoepidemiologic Profile of Cutaneous Leishmaniasis, Morocco

During the past 20 years, cutaneous leishmaniasis has emerged as a major public health threat in Morocco. We describe distribution of Leishmania major and L. tropica in Morocco and a new focus of cutaneous leishmaniasis due to L. infantum. We recommend using molecular techniques to diagnose suspected leishmaniasis cases.

CUTANEOUS LEISHMANIASIS: AN EMERGING INFECTION IN A NON-ENDEMIC AREA AND A BRIEF UPDATE

We report here the emergence of a new focus of cutaneous leishmaniasis (CL) due to Leishmania tropica (L. tropica) in the Ajmer city of Rajasthan, India, a previously non-endemic area. Between January-February 2006, 13 new indigenously acquired cases of CL were diagnosed among the patients attending the Skin and STD department, JLN Hospital, Ajmer. The diagnosis was based on clinical presentation, demonstration of amastigotes (LT bodies) in Giemsa stained smear of the lesion and response to intralesional / local anti-leishmanial drug therapy. In addition, culture of the promastigote forms of L. tropica from the lesion was successfully attempted in four of the smear negatives cases. By retrospective analysis, 23 new indigenous cases of CL have been diagnosed in the same setting during the period January 2004 - December 2005, based on clinical and therapeutic response alone. There was no clear-cut history of sandfly bite and travel outside the district or state to endemic area in any of the cases. However, all of them came from a common residential area (famous dargah of Ajmer) and the peak incidence was seen in January, four months after the famous Urs fair of Ajmer, the location was urban and the lesions were characteristic of L. tropica. Therefore, the disease is suspected to be anthroponotic. These features are suggestive of a common mode of transmission, source and/or vector signalling introduction of this infection into a non-endemic area.

Nyssomyia intermedia (Lutz & Neiva, 1912) and Nyssomyia neivai (Pinto, 1926) (Diptera: Psychodidae: Phlebotominae) geographical distribution and epidemiological importance

Nyssomyia intermedia (Lutz & Neiva 1912) and N. neivai (Pinto 1926) are possible vectors of tegumentary leishmaniasis in some regions of Brazil. Further, the latter was until recently, considered a junior synonym of the former. This study has the purpose of updating our knowledge of the geographical distribution of these species, based on specimens deposited at the collection of the Centro de Pesquisas René Rachou-Fiocruz, Faculdade de Saúde Pública-Universidade de São Paulo, and on data presented by literature as also to associate this distribution with the cutaneous leishmaniasis cases reported. It has been reported that N. intermedia occurs in the states of the Northeastern Region, in Rio de Janeiro, Espírito Santo, on the northern coast of São Paulo, in eastern Minas Gerais, Mato Grosso do Sul, and Goiás, close to the border with Minas Gerais and Bahia. N. neivai occurs in the Southern Region, southern coast and in western São Paulo, southern and western Minas Gerais, southern Goiás, and southern Pará, beyond Argentina, Bolivia, and Paraguay. It is important to highlight that N. intermedia and N. neivai occur in sympatry in Minas Gerais and São Paulo. N. intermedia or N. neivai are predominant or are captured abundantly in several cutaneous leishmaniasis foci in the Southeastern and Southern regions of Brazil.

CUTANEOUS LEISHMANIASIS CAUSED BY LEISHMANIA TROPICA IN BIKANER, INDIA: PARASITE IDENTIFICATION AND CHARACTERIZATION USING MOLECULAR AND IMMUNOLOGIC TOOLS

Identification of new foci of cutaneous leishmaniasis (CL), along with reports of Leishmania donovani causing the disease, is an issue of concern. Clinico-epidemiologic analysis of 98 cases in the endemic regions of Rajasthan state, India, suggested the preponderance of infection in men (62.24%) compared with women (37.75%). Species characterization by internal transcribed spacer 1 (ITS1) polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP), kDNA-PCR, and immunofluorescence assay established L. tropica as the causative organism. When applied directly to 32 clinical samples, kDNA PCR had a sensitivity of 96.6%, whereas ITS1 PCR had a sensitivity of 82.75%, thus facilitating diagnosis and species identification. Either parasite culture or direct microscopy alone detected 48.2% and 65.5% of the positive samples, respectively, whereas culture and microscopy together improved overall sensitivity to 89.3% (25/28). Except for the kDNA PCR, all other assays were 100% specific. This study provides the first comprehensive molecular and immunologic studies of CL in India.

MULTIPLE HYBRID GENOTYPES OF LEISHMANIA (VIANNIA) IN A FOCUS OF MUCOCUTANEOUS LEISHMANIASIS

The principal agent of mucocutaneous leishmaniasis (MCL) is the South American protozoan parasite Leishmania (Viannia) braziliensis. This organism is generally considered to be clonal, that is, it does not to undergo genetic exchange. Nevertheless, apparent hybrids between several Leishmania species have been reported in the New World and the Old World. When we characterized isolates of Leishmania (Viannia) from a single focus of cutaneous leishmaniasis (CL) and MCL, we found a remarkable phenotypic and genotypic diversity, with 12 zymodemes and 20 microsatellite genotypes. Furthermore, 26 of the 59 isolates were L. braziliensis/L. peruviana phenotypic hybrids that displayed 7 different microsatellite genotypes. A hybrid genotype was the only organism isolated from 4 patients with MCL. Thus hybrids must be included among the potential agents of MCL. Despite the propensity for clonality, hybrids are also an important feature of Leishmania (Viannia) and may give rise to epidemiologically important emergent genotypes.

Treating cutaneous leishmaniasis patients in Kabul, Afghanistan: cost-effectiveness of an operational program in a complex emergency setting

BackgroundAlthough Kabul city, Afghanistan, is currently the worldwide largest focus of cutaneous leishmaniasis (CL) with an estimated 67,500 cases, donor interest in CL has been comparatively poor because the disease is non-fatal. Since 1998 HealthNet TPO (HNTPO) has implemented leishmaniasis diagnosis and treatment services in Kabul and in 2003 alone 16,390 were treated patients in six health clinics in and around the city. The aim of our study was to calculate the cost-effectiveness for the implemented treatment regimen of CL patients attending HNTPO clinics in the Afghan complex emergency setting.MethodsUsing clinical and cost data from the on-going operational HNTPO program in Kabul, published and unpublished sources, and discussions with researchers, we developed models that included probabilistic sensitivity analysis to calculate ranges for the cost per disability adjusted life year (DALY) averted for implemented CL treatment regimen. We calculated the cost-effectiveness of intralesional and intramuscular administration of the pentavalent antimonial drug sodium stibogluconate, HNTPO's current CL 'standard treatment'.ResultsThe cost of the standard treatment was calculated to be US$ 27 (95% C.I. 20 – 36) per patient treated and cured. The cost per DALY averted per patient cured with the standard treatment was estimated to be approximately US$ 1,200 (761 – 1,827).ConclusionAccording to WHO-CHOICE criteria, treatment of CL in Kabul, Afghanistan, is not a cost-effective health intervention. The rationale for treating CL patients in Afghanistan and elsewhere is discussed.

Cutaneous leishmaniasis: An emerging infection in a non-endemic area and a brief update

We report here the emergence of a new focus of cutaneous leishmaniasis (CL) due to Leishmania tropica (L. tropica) in the Ajmer city of Rajasthan, India, a previously non-endemic area. Between January-February 2006, 13 new indigenously acquired cases of CL were diagnosed among the patients attending the Skin and STD department, JLN Hospital, Ajmer. The diagnosis was based on clinical presentation, demonstration of amastigotes (LT bodies) in Giemsa stained smear of the lesion and response to intralesional / local anti-leishmanial drug therapy. In addition, culture of the promastigote forms of L. tropica from the lesion was successfully attempted in four of the smear negatives cases. By retrospective analysis, 23 new indigenous cases of CL have been diagnosed in the same setting during the period January 2004 - December 2005, based on clinical and therapeutic response alone. There was no clear-cut history of sandfly bite and travel outside the district or state to endemic area in any of the cases. However, all of them came from a common residential area (famous dargah of Ajmer) and the peak incidence was seen in January, four months after the famous Urs fair of Ajmer, the location was urban and the lesions were characteristic of L. tropica. Therefore, the disease is suspected to be anthroponotic. These features are suggestive of a common mode of transmission, source and/or vector signalling introduction of this infection into a non-endemic area.

Distribution of Sand Fly (Diptera: Psychodidae) Species and Efficiency of Capturing Methods in Sanlıurfa Province, Turkey

The population dynamics of sand flies (Diptera: Psychodidae) were studied in Sanliurfa province in southeastern Turkey, in the country's largest focus of typical anthroponotic cutaneous leishmaniasis, during 2000-2002. Sand flies were collected at nine different sampling stations, located throughout the city, representing a cross section of urban and rural habitats. In total, 29,771 sand flies were collected, 45.35% of which were Phlebotomus papatasi Scopoli. In this study, the overall sand fly species diversity, relative abundance of each species, biodiversity, and similarity indices among sampling stations and efficiency of trapping methods were evaluated. Among the sampling stations, Sanliurfa city center and Suruç were shown to have the highest number of sand fly species; Harran-Akcakale and Hilvan habitats produced the largest number of individuals. The greatest similarity rates (80%) in terms of sand fly species were observed between Hilvan and city center, Harran-Akcakale and city center, Harran-Akcakale and Yenice, and Siverek and Viransehir. The lowest similarity rate (16%) was observed between Bozova-Birecik and city center. Comparison of biodiversity and similarity indices between the various sampling stations reveals the distribution of the suspected vector species and provides basic knowledge required to develop logical and effective control strategies. Among the trapping methods used, light traps showed the highest capture efficiency, above aspirators and sticky papers. It was concluded that light traps alone were sufficient to determine the sand fly fauna of the study area. It is recommended that the spatial and temporal dynamics of sand fly populations be monitored throughout the southeastern Anatolia Project (GAP) construction period, considering the potential impact the project may have on mean temperature, humidity, and human population movements.

Distribution of Sand Fly (Diptera: Psychodidae) Species and Efficiency of Capturing Methods in Sanlıurfa Province, Turkey

The population dynamics of sand flies (Diptera: Psychodidae) were studied in Sanliurfa province in southeastern Turkey, in the country's largest focus of typical anthroponotic cutaneous leishmaniasis, during 2000-2002. Sand flies were collected at nine different sampling stations, located throughout the city, representing a cross section of urban and rural habitats. In total, 29,771 sand flies were collected, 45.35% of which were Phlebotomus papatasi Scopoli. In this study, the overall sand fly species diversity, relative abundance of each species, biodiversity, and similarity indices among sampling stations and efficiency of trapping methods were evaluated. Among the sampling stations, Sanliurfa city center and Suruç were shown to have the highest number of sand fly species; Harran-Akcakale and Hilvan habitats produced the largest number of individuals. The greatest similarity rates (80%) in terms of sand fly species were observed between Hilvan and city center, Harran-Akcakale and city center, Harran-Akcakale and Yenice, and Siverek and Viransehir. The lowest similarity rate (16%) was observed between Bozova-Birecik and city center. Comparison of biodiversity and similarity indices between the various sampling stations reveals the distribution of the suspected vector species and provides basic knowledge required to develop logical and effective control strategies. Among the trapping methods used, light traps showed the highest capture efficiency, above aspirators and sticky papers. It was concluded that light traps alone were sufficient to determine the sand fly fauna of the study area. It is recommended that the spatial and temporal dynamics of sand fly populations be monitored throughout the southeastern Anatolia Project (GAP) construction period, considering the potential impact the project may have on mean temperature, humidity, and human population movements.

Estudo de dois anos com animais reservatórios em área de ocorrência de leishmaniose tegumentar americana humana em bairro de urbanização antiga na cidade de Manaus-AM, Brasil

Os autores descrevem o resultado de dois anos de investigação de um foco de Leishmaniose Tegumentar Americana (LTA), ocorrida em área urbanizada em um conjunto habitacional na cidade da Manaus-AM. Chama atenção o fato de que este não é o padrão de ocorrência de surtos da doença na região, e sim interrupção da transmissão após urbanização. Foram investigados os animais considerados reservatórios em potencial para a leishmaniose em domicílios humanos e em áreas de floresta adjacentes. Foram testados anticorpos contra Leishmania spp em amostras de sangue de cães e detectada reatividade pela reação de imunofluorescencia indireta em oito (20,51 %) dos examinados. Entre os animais silvestres examinados a espécie Didelphis marsupialis foi predominante, com 20 exemplares capturados, sendo encontrados homoflagelados em três destes e lesões suspeitas de leishmaniose cutanea em dois. Acredita-se que um assentamento populacional desordenado ocorrido nas adjacências tenha causado o deslocamento das populações de vetores e reservatórios naturais em direção às casas do conjunto Hiléia propiciando o surto.

Bionomics of phlebotomine sandflies at a peacekeeping duty site in the north of Sinai, Egypt

A longitudinal entomological survey for sandflies was conducted from 1989 to 1991 at a focus of enzootic cutaneous leishmaniasis in Northeast Sinai, Egypt, within the border region monitored by multinational peacekeepers. Standardized sampling with CDC light traps, oiled paper “sticky traps”, and human landing collection was employed to determine monthly trends in species composition, density, sex ratio, and reproductive status of vector sandflies. Each collection method independently defined sandfly seasonality as the period May–November in 1990, and March–October in 1991. Plebotomus papatasi was the only anthropophagic species found and comprised more than 94% of the sandfly population. Two population peaks (May, July) were observed for this species in both survey years. Density of P. papatasi in underground bunkers was higher than outside but inflated by a greater proportion of male flies. During 1990, the proportion of gravid P. papatasi increased progressively during the 5 months period from May to September and averaged 29.5% and 29.7% for interior and exterior collections, respectively. Density of P. papatasi was greater during 1991, but proportions of gravid flies were significantly lower in each survey month and averaged 14.9% and 12.3% for interior and exterior collections, respectively. Seasonal rates of Leishmania-infected P. papatasi averaged 0.8% and 0.9% in 1989 and 1990, but fell to zero in 1991, suggesting an unstable focus of Leishmania major transmission. Proportions of gravid flies may be a valid indicator of the physiological age and epidemiologic importance of the vector sandfly population at this focus. The strong correlation of sticky trap indices to human-landing/biting rates shows that this is an accurate, inexpensive, and no-risk alternative to human bait collections.

Natural infection of Phlebotomus ( Larroussius) langeroni (Diptera: Psychodidae) with Leishmania infantum in Tunisia

Phlebotomine sand flies were captured from an active transmission focus of sporadic cutaneous leishmaniasis, caused by Leishmania infantum, in El Kef region, northern Tunisia. Both Phlebotomus perniciosus and P. langeroni were found. Phlebotomus langeroni females showed a statistically significant intradomiciliary dominance (P<0.01 for the 2003 and 2004 seasons) when compared to animal shelters. During the 2003 season, dissection of collected female specimens showed the presence of flagellates within the digestive tracts of two P. perniciosus among 1086 observed, but none in 232 P. langeroni. Amplification of kinetoplast minicircles of Leishmania parasites was applied to DNA samples extracted from 298 frozen females including 249 P. perniciosus, 36 P. langeroni, 5 P. longicuspis and 8 P. perfiliewi and revealed by radioactive probe hybridization. Two P. langeroni females showed a signal of the size expected for L. infantum (800bp) indicating infection with these parasites. However, this PCR-hybridization method failed to identify any positive P. perniciosus females in pools of specimens. These results show for the first time the natural infection of P. langeroni with L. infantum in Tunisia, and support the existence of different L. infantum transmission cycles in Tunisia, with a potential role for P. langeroni as a vector.

Isolation ofLeishmania (Viannia) braziliensisfromLutzomyia spinicrassa(species groupVerrucarum) Morales Osorno Mesa, Osorno &amp; Hoyos 1969, in the Venezuelan Andean Region

Natural infection with Leishmania spp. in phlebotomine sandflies was searched for during a longitudinal study carried out from July 1997 to July 1998 in the village Catarnica, Municipality Independencia, Táchira State. This hamlet is an old endemic focus of cutaneous leishmaniasis in the Venezuelan Andean region, which lies close to the Colombian border at 1,300 m a.s.l., in an agricultural area mainly used for cultivating coffee. Phlebotomine sandflies were collected using Shannon traps placed in the peridomestic habitat from 19:00 to 21:00 hs. Males were stored in alcohol 70 % while females were kept in Nunc vials with 10 % DMSO and cryopreserved in liquid nitrogen for subsequent dissection and identification. The most abundant anthropophilic species was Lutzomyia spinicrassa with 3,032 males and 4,290 females (85.4%). Among 1,633 (38%) females of Lu. spinicrassa dissected, 26 11.6%) were infected with promastigotes, while no natural infection was found in 209 females of other species. The flagellates were identified as Leishmania braziliensis braziliensis using PCR with species specific primers derived from nuclear DNA and hybridization using species specific probe labelled with digoxigenin. This parasite had been previously isolated from patients with cutaneous leishmaniasis from the same area. These results show Lu. spinicrassa as a new proven vector of Leishmania braziliensis in the Andean region of Venezuela.