Species Of Phlebotomine Sandflies Research Articles

Species-specific antimonial sensitivity in Leishmania is driven by post-transcriptional regulation of AQP1.

Leishmania is a digenetic protozoan parasite causing leishmaniasis in humans. The different clinical forms of leishmaniasis are caused by more than twenty species of Leishmania that are transmitted by nearly thirty species of phlebotomine sand flies. Pentavalent antimonials (such as Pentostam or Glucantime) are the first line drugs for treating leishmaniasis. Recent studies suggest that pentavalent antimony (Sb(V)) acts as a pro-drug, which is converted to the more active trivalent form (Sb(III)). However, sensitivity to trivalent antimony varies among different Leishmania species. In general, Leishmania species causing cutaneous leishmaniasis (CL) are more sensitive to Sb(III) than the species responsible for visceral leishmaniasis (VL). Leishmania aquaglyceroporin (AQP1) facilitates the adventitious passage of antimonite down a concentration gradient. In this study, we show that Leishmania species causing CL accumulate more antimonite, and therefore exhibit higher sensitivity to antimonials, than the species responsible for VL. This species-specific differential sensitivity to antimonite is directly proportional to the expression levels of AQP1 mRNA. We show that the stability of AQP1 mRNA in different Leishmania species is regulated by their respective 3’-untranslated regions. The differential regulation of AQP1 mRNA explains the distinct antimonial sensitivity of each species.

Exploring the utility of phylogenetic analysis of cytochrome oxidase gene subunit I as a complementary tool to classical taxonomical identification of phlebotomine sand fly species (Diptera, Psychodidae) from southern Europe

Phlebotomine sand flies (Diptera, Psychodidae) are known to be vectors of several pathogens such as Leishmania and Phlebovirus genera. The identification of phlebotomine sand fly species is currently based on morphological characters, and requires considerable taxonomic expertise and skilfulness, but may be complemented by DNA-based analyses for (i) accurate species identification and (ii) for estimating sand fly diversity. The aim of this study was to evaluate the utility of mitochondrial cytochrome oxidase gene subunit I (cox1) sequence analysis as a complementary tool to classical taxonomical for the identification of the most prevalent phlebotomine sand fly species from southern Europe (i.e. Phlebotomus ariasi, P. perniciosus, P. sergenti and Sergentomyia minuta).Phylogenetic analyses of cox1 sequences allowed conclusive assignment of most of the sand flies into individual species, and revealed the genetic heterogeneity that characterizes some of the identified genetic clusters. Nevertheless, it showed some limitations, as it failed to (i) allocate correctly all of all species of a given subgenus to a single lineage, or (ii) conclusively identify sequences amplified from individuals classified morphologically as P. ariasi. A more extensive analysis of cox1 sequences together with morphometric characterization of specimens from different geographic areas/regions might be useful for the correct assessment of the phylogenetic relationship within the P. ariasi/P. chadlii cluster and/or help to ascertain the usefulness of cox1 for molecular taxonomy of sand flies.

Morphological keys for the identification of Italian phlebotomine sand flies (Diptera: Psychodidae: Phlebotominae).

BackgroundPhlebotomine sand flies are small blood-feeding insects of great medical and veterinary significance. Their identification relies basically on the microscopic examination of key morphological characters. Therefore, identification keys are fundamental to any researcher dealing with these insects. The Italian fauna of phlebotomine sand flies consists of eight species (Phlebotomus perniciosus, Phlebotomus perfiliewi, Phlebotomus ariasi, Phlebotomus neglectus, Phlebotomus papatasi, Phlebotomus mascittii, Phlebotomus sergenti and Sergentomyia minuta), whose morphological delineation may be troublesome for non-taxonomists.MethodsA total of 8,757 pictures were taken from the 419 selected phlebotomine sand fly specimens collected on different occasions. Twenty-eight characters for the males and 23 for the females were examined, resulting in a database containing over 10,000 entries. Representative phlebotomine sand fly specimens for each species available were selected and relevant characters were drawn with the aid of a camera lucida.ResultsAfter detailed morphological study of representative specimens, comprehensive identification keys based on key characters (e.g., pharynx and spermathecae of females and male terminalia) were elaborated.ConclusionsThe identification keys provided herein allow the identification of genera and species of phlebotomine sand flies of Italy and they will facilitate future studies on these medically important insects.

Description of Trichophoromyia uniniensis, a new phlebotomine species (Diptera: Psychodidae: Phlebotominae) of Amazonas State, Brazil.

BackgroundA new species of phlebotomine sand flies belonging to Trichophoromyia Barretto, 1962 genus is described, based on males collected in Jaú National Park, Amazonas state, Brazil.MethodsThe Sand flies were mounted in Canada balsam. They were measured with a binocular Olympus CH-2 microscope with the aid of a micrometer objective and the drawings were done with the help of a camera lucida.ResultsThis new species named Trichophoromyia uniniensis sp. nov. is closely related to Trichophoromyia omagua (Martins, Llanos & Silva, 1976). The former can be distinguished from the latter by the shape of its paramere that has the lower apical region turned up in the new species.ConclusionWith the new species here described a total of 39 species belonging to the Trichophoromyia genus are now known, most of them present in the Amazon rainforest.

Molecular detection of Leishmania in phlebotomine sand flies in a cutaneous and visceral leishmaniasis endemic area in northeastern Brazil.

Several phlebotomine sand fly species have been regarded as putative or proven vectors of parasites of the genus Leishmania in Brazil, but data for the northeastern region remains incipient. In this study, a total of 600 phlebotomine sand flies were grouped in pools of 10 specimens each and tested by a Leishmania genus-specific PCR and by a PCR targeting Leishmania (Leishmania) infantum. Fourteen out of 60 pools were positive by the genus-specific PCR, being five pools of L. migonei, seven of L. complexa, one of L. sordellii and one of L. naftalekatzi, which correspond to a minimal infection rate of 2.3% (14/600). Our results, associated with their known anthropophily and their abundance, suggest the participation of L. migonei and L. complexa as vectors of Leishmania in northeastern Brazil. Remarkably, this is the first time in this country that the detection of Leishmania DNA in L. sordellii and L. naftalekatzi has been reported, but future studies are necessary to better understand the significance of these findings.

Lista actualizada de flebotomíneos (Diptera: Psychodidae: Phlebotominae) de la región cafetera colombiana

An updated list of phlebotomine sand flies species in coffee growing areas in the Colombian Andean region is presented. Fifty three species were reported from 12 departments. In addition, species distribution in the region was derived from specimens obtained during intensive field work in five departments, from previously published studies and from the taxonomic revision of specimens in the entomological collection of the Programa de Estudio y Control de Enfermedades Tropicales (PECET). The list includes the genera Brumptomyia (2 species), Lutzomyia (50 species) and Warileya (1 species). The updated list contains eleven new records in the region under study, including Lutzomyia panamensis , a species of medical importance not recorded previously in this zone. Eighteen of the species are considered to be anthropophilic, and many of them have been implicated in the transmission of leishmaniasis.

Morphological keys for the identification of Italian phlebotomine sand flies (Diptera: Psychodidae: Phlebotominae)

Phlebotomine sand flies are small blood-feeding insects of great medical and veterinary significance. Their identification relies basically on the microscopic examination of key morphological characters. Therefore, identification keys are fundamental to any researcher dealing with these insects. The Italian fauna of phlebotomine sand flies consists of eight species (Phlebotomus perniciosus, Phlebotomus perfiliewi, Phlebotomus ariasi, Phlebotomus neglectus, Phlebotomus papatasi, Phlebotomus mascittii, Phlebotomus sergenti and Sergentomyia minuta), whose morphological delineation may be troublesome for non-taxonomists. A total of 8,757 pictures were taken from the 419 selected phlebotomine sand fly specimens collected on different occasions. Twenty-eight characters for the males and 23 for the females were examined, resulting in a database containing over 10,000 entries. Representative phlebotomine sand fly specimens for each species available were selected and relevant characters were drawn with the aid of a camera lucida. After detailed morphological study of representative specimens, comprehensive identification keys based on key characters (e.g., pharynx and spermathecae of females and male terminalia) were elaborated. The identification keys provided herein allow the identification of genera and species of phlebotomine sand flies of Italy and they will facilitate future studies on these medically important insects.

Phlebotomine sand flies from Madagascar (Diptera: Psychodidae). VIII — Phlebotomus (Anaphlebotomus) vincenti n. sp.

A new species of Phlebotomine sand fly (Diptera: Psychodidae) is described from Mikea Forest in the southwest of Madagascar: Phlebotomus vincenti n. sp. It is temporarily classified in the subgenus Anaphlebotomus according to its spermathecal organization, similar to those of P. berentiensis, P. fertei and P. vaomalalae, previously classified in the subgenus Anaphlebotomus referring to on male genitalia morphology. An original trumpet-like dilatation on the distal part of the spermathecal ducts individualizes P. vincenti n. sp. from the other Malagasy species. Moreover, the second internal transcribed spacer (ITS2) sequences strongly individualises P. vincenti n. sp. from the other Malagasy Phlebotomus.

Editorial Commentary: Local Treatments for Cutaneous Leishmaniasis

Cutaneous leishmaniasis (CL) is an infection caused by various species of Leishmania protozoa, which are usually transmitted by the bite of various species of phlebotomine sandflies [1]. It is a “neglected tropical disease” that causes significant morbidity and social stigmatization and also occurs in subtropical regions such as southern Europe and the southern United States. Its clinical presentations and natural history vary considerably, and although nearly all cases will eventually heal spontaneously, this can take more than a year and be complicated by secondary bacterial infections, lymphatic involvement, local recurrences, and mucosal leishmaniasis due to Leishmania (Viannia) braziliensis complex infections in certain New World countries. Laboratory diagnosis requires specialist parasitology and molecular biology techniques, and numerous local and systemic treatment options are available. Recent studies [2, 3] offer a timely reminder of the value of local (topical or intralesional) treatments for CL, when most research is focused on systemic (oral or parenteral) treatments instead. Morizot et al [2] show how national treatment guidelines and expert advice can optimize the use of simple wound care only or local treatment for a wide range of CL infections in travelers with impressive results. Overall, 62% of cases were treated this way and cure rates within 60 days of starting treatment were 92% for wound care only and 79% for cryotherapy plus intralesional antimony, compared to 60% for systemic treatments. The majority of cases had a good prognosis and were suitable for nonsystemic treatment according to the national guidelines, whereas those with a worse prognosis were more likely to be given systemic treatment, and so there is obvious selection bias. However, some centers would have treated all of these cases with systemic treatments at higher cost and with more adverse effects and so the results remain valuable. Interestingly, no patients seem to have withdrawn due to the transient but severe pain sometimes caused by intralesional treatment (or other forms of cryotherapy) that may dissuade physicians from using such techniques. Systemic treatments for CL are often preferred for several reasons, including the fact that most new therapies for CL are derived from those used for visceral leishmaniasis, which are inevitably systemic in nature. Local treatments are also more difficult to evaluate due to problems in standardizing the dose given during administration of a topical or intralesional drug. Even when local treatments are known to be effective, there may be problems in the provision of therapies such as intralesional pentavalent antimonial (SbV) drugs (sodium stibogluconate or meglumine antimoniate), cryotherapy, topical paromomycin (aminosidine), and thermotherapy. Overall, our healthcare systems often make it

Detection of Leishmania infantum and identification of blood meals in Phlebotomus perniciosus from a focus of human leishmaniasis in Madrid, Spain

Since 2010, the number of cases of both human visceral leishmaniasis and cutaneous leishmaniasis in southwestern Madrid region (Spain) and more specifically in the town of Fuenlabrada has increased. Direct xenodiagnosis of leishmaniasis proved that hares (Lepus granatensis) from this focus are able to infect with Leishmania infantum colonized Phlebotomus perniciosus. To a better understanding of this focus of leishmaniasis, we conducted an entomological survey using CDC light traps, at the end of the seasonal transmission period of 2011 before the beginning of control measures of the disease, to study the phlebotomine sand flies species involved. Detection of Leishmania DNA in the sand flies captured was studied by kDNA-PCR and cpb-PCR. In addition, blood fed and gravid female P. perniciosus were analysed by a PCR based in vertebrate cytochrome b (cyt b) gene. Taxonomic identification of captured sand flies (n = 174) as P. perniciosus (n = 171) and Sergentomyia minuta (n = 3) together with the analysis of blood feeding in ten sand flies that shows a high preference for hares (n = 6), followed by humans (n = 3), and cats (n = 1) confirm a strong association between P. perniciosus hares and humans in the focus. Moreover, 79 out of 135 (58.5 %) P. perniciosus were positive to L. infantum by PCR approaches. These data support the increase of human leishmaniasis cases in the area and the existence of an unusual sylvatic cycle alternative to the classical domestic one, where the dog is the main reservoir of L. infantum.

Ocorrência de flebotomíneos (Diptera: Psychodidae: Phlebotominae) no ambiente peridomiciliar em área de foco de transmissão de leishmaniose tegumentar no município de Manaus, Amazonas

A leishmaniose é uma antropozoonose com focos naturais, nos quais a infecção é mantida em ecótopos silvestres entre a população de vetores e seus hospedeiros vertebrados. Neste estudo foram identificadas espécies de flebotomíneos envolvidas na transmissão de leishmaniose tegumentar no Bairro Cidade de Deus, onde há focos da doença humana e animal. O bairro está situado à margem da reserva florestal Adolpho Ducke. Foram capturados 122 espécimens em épocas variadas, utilizando-se armadilhas luminosas do tipo CDC. As espécies de flebotomíneos mais encontradas foram Lutzomyia umbratilis, L. rorotaensis e L. olmeca nociva. Nas proximidades de uma das residências onde foram realizadas as coletas de flebotomíneos, verificou-se descarte de resíduos orgânicos (lixo), o que possibilita a atração de reservatórios silvestres presentes na reserva florestal favorecendo a ocorrência do ciclo de transmissão da doença no peridomicílio.

Tetracycline and trimethoprim/sulfamethoxazole at clinical laboratory: can they help to characterize Staphylococcus aureus carrying different SCCmec types?

This work was carried out on the purpose of identifying the species of phlebotomine sandflies in the municipality of Monte Negro, state of Rondonia, Brazil, that may have been transmitting the American cutaneous leishmaniasis (ACL), and concisely describe epidemiological aspects of disease. The epidemiologic and socioeconomical indicators were obtained from government institutions and the local Municipal Secretary of Health. Phlebotomine sandflies were captured using CDC light traps between July 2006 to July 2008. The total of 1,240 of female sandflies were examined by PCR method directed to k-DNA. There has been a significant decrease in the incidence of ACL of about 50% over the last ten years in the municipality. A total of 1,935 specimens of 53 sandfly species were captured, three of the genus Brumptomyia genus and 50 of the genus Lutzomyia. The predominant species was Lutzomyia acanthopharynx, Lutzomyia whitmani, Lutzomyia geniculata and Lutzomyia davisi. None were positive for Leishmania sp. Four sandflies species were found in the State of Rondonia for the first time: Brumptomyia brumpti, Lutzomyia tarapacaensis, Lutzomyia melloi and Lutzomyia lenti. The presence of Lutzomyia longipalpis, was also captured. Socioeconomical improvement of Brazilian economy and the increase of environmental surveillance in the last 15 years collaborated in the decrease of people exposed to vectors, reducing the incidence of ACL.

Changes in Phlebotomine Sand Fly Species Composition Following Insecticide Thermal Fogging in a Rural Setting of Western Panamá

American Cutaneous Leishmaniasis, ACL, is a zoonotic disease with a large richness of co-occurring vector species in transmission foci. Here, we describe changes in patterns of phlebotomine sand fly (Diptera: Psychodidae: Phlebotominae) species composition at the village of Trinidad de Las Minas, Capira, Panamá, a hyperendemic focus of ACL transmission, subjected to a vector control intervention with insecticide thermal fogging (ITF). Our study setting consisted of 24 houses, 12 subjected to two rounds of ITF and 12 kept as control. During 15 months (April 2010– June 2011) we monitored sand fly species composition and abundance with modified HP light traps inside (domicile) and outside (peridomicile) the studied houses. From 5628 sand flies collected, we were able to identify 5617 of the samples into 24 species, a number of species close to 25±1.6, the estimate from the Chao2 Index. The most abundant species were Lutzomya trapidoi (20%), Lu. gomezi (20%) and Lu. triramula (20%). Cluster analyses showed that most of the 24 houses had high similarity in relative abundance patterns of the six most common species, with only few peripheral houses not following the main cluster pattern. We also found that species richness was decreased to 22 species in the fogged houses, of which only 19 were found in the domiciliary environment. Changes in species richness were especially notorious at the end of the wet season. Our results suggest that species richness can decrease following ITF in domiciliary environments, primarily affecting the less common species.

Infectividad del perro (Canis familiaris) para Lutzomyia youngi en Trujillo, Venezuela

In Trujillo, Venezuela the prevalence for American tegumentary leishmaniasis (ATL) is 38 per 100,000 inhabitants. In a periurban, rural settlement of the capital city Trujillo, we studied the potential capability of the domestic dog (Canis familiaris) as a source of infection for Lutzomyia youngi, a phlebotomine sand fly species abundant in the study area and whose domestic vectorial activity has been proven. Dogs with dermal lesions suggestive of ATL and parasitological confirmation of infection, were selected for xenodiagnosis by allowing sylvatic phlebotomines from a ATL free area, to feed ad libitum over each animal's entire body surface. The insects' intestinal tracts were dissected 5 days after the blood meal in order to look for flagellate forms. When these were found, parasitological identification was performed by the multiplex-PCR technique. Four hundred and fifty five sand flies engorged over two dogs in three different assays; promastigotes were found in 4 (0.88%) of the specimens on only one occasion. PCR identified DNA of the Leishmania Viannia subgenus. The household dog has the potential of being a domestic risk factor in the ATL transmission cycle.

Bilateral Anomaly in Evandromyia evandroi (Diptera: Psychodidae: Phlebotominae) Captured in Vicência Municipality, Northern Rainforest Region of Pernambuco State, Brazil

Approximately 800 species of phlebotomine sand flies, many of which are vectors of Leishmania, have been described. Besides morphological similarities within groups, the occurrence of anomalies within a species may lead to an erroneous description of new species. This paper describes one phlebotomine sand fly, Evandromyia evandroi, with a symmetrical bilateral anomaly in the number of spines on the gonostyle. In this specimen, the anomalous spine is located in the external region of gonostyle, inserted between the upper external and the lower external spines. It is important to document morphological anomalies, so as to avoid erroneous sand fly identifications.

A New Species ofEvandromyia(Diptera: Psychodidae: Phlebotominae) From Minas Gerais State, Brazil

A new species of phlebotomine sand fly found in Minas Gerais state, Brazil, is described and illustrated based on male and female morphological characteristics. The new species belongs to the rupicola series within the subgenus Evandromyia Mangabeira. Both sexes are morphologically close to Evandromyia rupicola (Martins, Godoy & Silva) and Evandromyia grimaldii Andrade Filho, Pinto, Santos & Carvalho and the characteristics for their classification are here presented.

Morphometric and molecular characterization of the series Guyanensis (Diptera, Psychodidae, Psychodopygus) from the Ecuadorian Amazon Basin with description of a new species

Several populations of the series Guyanensis (Diptera, Psychodidae, Psychodopygus) were collected from the Yasuní National Park in the Ecuadorian Amazon region. The specimens comprised the species Psychodopygus geniculatus, Psychodopygus luisleoni and Psychodopygus corossoniensis. Within Ps. geniculatus, we observed two populations, one with a narrow paramere and relatively short genital filaments and the other characterized by a wider coxite and longer genital filaments. A multiple approach combining morphology, morphometry and DNA sequencing of the ribosomal internal transcribed spacer 2 (ITS2) and the mitochondrial cytochrome b gene was carried out. Morphological, morphometric and molecular data strongly suggested the presence of two populations within Ps. geniculatus. The lack of intermediate forms within these populations supported the proposal of two sympatric species. This report describes Psychodopygus francoisleponti n. sp.

Lista de espécies de Phlebotominae (Diptera, Psychodidae) do Estado de São Paulo, Brasil, com comentários sobre sua distribuição geográfica

Os flebotomíneos são insetos de interesse médico, responsáveis pela transmissão de protozoários parasitas do gênero Leishmania entre animais reservatórios humanos e não-humanos, os quais são encontrados em todo o Estado de São Paulo, Brasil. As 69 espécies de flebotomíneos registradas no Estado de São Paulo, incluindo 7 espécies registradas aqui pela primeira vez, estão organizadas em uma lista de espécies utilizando a classificação filogenética de Galati (2003). Nossa lista de espécies incorpora e atualiza os dados publicados por Barretto (1947) e Martins et al. (1978), e inclui registros para mais 33 espécies retirados da literatura publicada desde então, e também da observação de espécimes depositados em coleções entomológicas. Para cada espécie de flebotomíneo, a distribuição geográfica por município é fornecida, com comentários sobre a distribuição de seis vetores de leishmaniose cutânea, bem como de Lutzomyia longipalpis, o principal vetor de leishmaniose visceral.

Towards a rapid molecular identification of the common phlebotomine sand flies in the Mediterranean region

The present study reports two simple molecular approaches allowing a rapid identification of the most prevalent species of phlebotomine sand flies in the Mediterranean region. A PCR protocol for the amplification of ITS2 ribosomal region and a PCR-RFLP on a mitochondrial DNA fragment (cytb–nd1) were settled in order to identify and discriminate among Phlebotomus perniciosus, Phlebotomus neglectus, Phlebotomus perfiliewi, Phlebotomus papatasi and Sergentomyia minuta. The ITS2 regions showed a certain degree of interspecific variability, which led to PCR amplicons of different sizes, i.e., 450, 490, 460, 480 and 530bp for P. perniciosus, P. neglectus, P. perfiliewi, P. papatasi, and S. minuta, respectively. Analogously, the digestion of a mitochondrial DNA amplicon with Ase I enzyme showed five different restriction profiles, which allowed the unequivocal differentiation of the sand fly species examined. These methods might represent useful tools for a molecular large scale screening of phlebotomine sand fly species caught in areas where leishmaniasis is endemic, in order to plan appropriate epidemiological surveillance programs for both Leishmania spp. and their vectors.

The sand fly fauna (Psychodidae: Phlebotominae) in the region of Saquarema, State of Rio de Janeiro, Brazil, an endemic area of cutaneous leishmaniasis transmission

Cutaneous leishmaniasis, caused by Leishmania (Viannia) braziliensis, is sporadic in many rural and suburban areas of Rio de Janeiro State. An investigation was carried out during 2008/9 in the Municipality of Saquarema, Rio de Janeiro, Southeast Brazil, in order to identify the phlebotomine sand fly fauna. More than 2,100 sand flies were collected in peridomestic areas in two chicken coops using CDC light traps. Nine species of phlebotomine sand flies were identified: Nyssomyia intermedia, Nyssomyia whitmani, Pintomyia (P.) pessoai, Pintomyia (P.) fischeri, Pintomyia (P.) bianchigalatiae, Migonemyia (M.) migonei, Lutzomyia (L.) longipalpis, Brumptomyia cunhai and Brumptomyia guimaraesi. Based on the results of this study together with related studies in other CL foci in Rio de Janeiro, both Nissomyia intermedia and Migonemyia migonei can be considered suspect vectors of the disease in the region. The potential risk of VL due to the presence of its proven vector L. longipalpis is discussed.