Peridomestic natural breeding sites of Nyssomyia whitmani (Antunes and Coutinho) in an endemic area of tegumentary leishmaniasis in northeastern Argentina

The scarce information about breeding sites of phlebotomines limits our understanding of the epidemiology of tegumentary leishmaniasis. Identifying the breeding sites and seasons of immature stages of these vectors is essential to propose prevention and control strategies different from those targeting the adult stage. Here we identified the rural breeding environments of immature stages of Ny. whitmani, vector species of Leishmania braziliensis in the north of Misiones province, Argentina; then we determined and compared the environmental and structural characteristics of those sites. We also identified the season of greatest emergence and its relationship with adult abundance. During a first collection period, between 28 and 48 emergence traps were set continuously for 16 months in six environments of the farm peridomicile and domicile: below house, chicken shed, experimental chicken shed, forest edge, pigsty and under fruit tree. Traps were checked and rotated every 40 nights. A total of 146 newly emerged individuals were collected (93.8% of them were Ny. whitmani), totaling an effort of 23,040 emergence trap-nights. The most productive environments were chicken shed and below house, and the greatest emergence was recorded in spring and summer. During a second collection period, emergence traps and light traps for adult capture were placed in the chicken shed and below house environments of eight farms. Emergence traps were active continuously during spring, summer, and early autumn. Environmental and structural characteristics of each environment were recorded. A total of 84 newly emerged phlebotomines (92.9% Ny. whitmani; 72,144 emergence trap-nights) and 13,993 adult phlebotomines (147 light trap-nights) were recorded in the chicken shed and below house environments. A positive correlation was also observed between trap success of newly emerged phlebotomines and of adults after 120 days. A high spatial variability was observed in the emergence of Ny. whitmani, with the number of newly emerged individuals being highest in soils of chicken sheds with the highest number of chickens and closest to forest edge. Moreover, below house was found to be as important as chicken sheds as breeding sites of Ny. whitmani. Management of the number of chickens in sheds, soil moisture and pH, and the decision of where to localize the chicken sheds in relation to the houses and the forest edge, might contribute to reduce the risk of human vector exposure and transmission of Leishmania.     

Sandflies (Diptera: Psychodidae) in rural and urban environments in an endemic area of cutaneous leishmaniasis in southern Brazil

The high proportion of cases of cutaneous leishmaniasis reported amongst residents in the city of Bandeirantes, in the state of Paraná, Brazil, led the authors to investigate the phlebotomine fauna in both urban and rural environments. The sandflies were captured with automatic light traps from 07:00 pm-07:00 am fortnightly in 11 urban peridomiciles from April 2008-March 2009 and monthly in three ecotopes within four rural localities from April 2009-March 2010. In one of these latter localities, sandfly capture was conducted with white/black Shannon traps during each of three seasons: spring, summer and fall. A total of 5,729 sandflies of 17 species were captured. Nyssomyia neivai (46.7%) and Nyssomyia whitmani (35.3%) were the predominant species. In this study, 3,865 specimens were captured with automatic light traps: 22 (0.083 sandflies/trap) in the urban areas and 3,843 (26.69 sandflies/trap) in the rural areas. Ny. neivai was predominant in urban (68.2%) and rural (42.8%) areas. A total of 1,864 specimens were captured with the white/black Shannon traps and Ny. neivai (54.5%) and Ny. whitmani (31.4%) were the predominant species captured. The small numbers of sandflies captured in the urban areas suggest that the transmission of Leishmania has occurred in the rural area due to Ny. neivai and Ny. whitmani as the probable vectors.     

Distribution of phlebotomine fauna (Diptera: Psychodidae) across an urban-rural gradient in an area of endemic visceral leishmaniasis in northern Brazil

The number of visceral leishmaniasis (VL) cases has increased over the past 10 years in Brazil, especially in the North and Northeast regions of the country. The aim of this study was to evaluate the urbanisation of VL vectors in Barcarena, Pará, an area in northern Brazil where VL is endemic. Sandflies were captured using Centers for Disease Control (CDC) light traps along an urban-rural gradient. The CDC traps were installed inside hen houses at a height of 150 cm. A total of 5,089 sandflies were collected and 11 species were identified. The predominant species was Lutzomyia longipalpis (rate of 95.15%), which suggests its participation in the transmission of VL. A total of 1,451 Lu. longipalpis females were dissected and no Leishmania infections were detected. Most of the sandflies were captured at the border of a forest (88.25%) and no flies were captured in the urban area, which suggests that transmission is still restricted to rural sites. However, the fact that a specimen was collected in an intermediate area indicates that urbanisation is a real possibility and that vector monitoring is important.     

Phlebotomine fauna (Diptera: Psychodidae) and species abundance in an endemic area of American cutaneous leishmaniasis in southeastern Minas Gerais, Brazil

This study was undertaken to identify the phlebotomine fauna and species abundance in domiciliary and peridomiciliary (hen-house and guava-tree) environments, on a lake shore, a cultivated area of coffee and banana, and a forested area of Concei??o da Aparecida municipality, southeastern the state of Minas Gerais, to provide information for the control and epidemiological surveillance of leishmaniasis in this area. The captures were carried out monthly between May 2001 and November 2002, with automatic light and Shannon traps. A total of 1444 sand flies were captured, 951 (76.5%) with automatic light traps and 493 (23.5%) with the Shannon trap. Thirteen species were captured, the most frequent being Nyssomyia whitmani (62.7%), Migonemyia migonei (21.4%), Pintomyia fischeri (6.9%), and Evandromyia lenti (3.6%). Species abundance was determined using the automatic light traps installed in the six environments. The most abundant species according to the standardized index of species abundance were Ny. whitmani (1.0) and Mg. migonei (0.82). In view of the dominance of these two species, known vectors of cutaneous leishmaniasis in other Brazilian areas, their participation in the transmission of the disease in this county is suggested. The diversity and evenness indexes in the domicile were the lowest due to the high frequency (83%) of Ny. whitmani. The capture of Lutzomyia longipalpis, rarely recorded in the south-eastern and southern regions of Minas Gerais, is also noteworthy.     

Dispersal pattern of the sand fly Lutzomyia neivai (Diptera: Psychodidae) in a cutaneous leishmaniasis endemic rural area in Southeastern Brazil

The dispersal pattern of the sand fly Lutzomyia neivai was studied through mark-release-recapture experiments in an American cutaneous leishmaniasis endemic rural area in Southeastern Brazil. Over 6500 specimens were marked with fluorescent powder and released in forest edge and peridomicile habitats from August to November 1999, February and April 2000. Recapture attempts were made using Shannon and CDC traps up to eight successive nights after releases. A total of 493 (7.58%) specimens were recaptured. The number of recaptured males and females of L. neivai in CDC traps was not affected by the distance between the trap and the release points. Approximately 90% of males and females recaptured in CDC traps were caught up to 70 m from the release points. The maximum female flight range recorded was 128 m. The average flight range per day was less than 60 m for males and females. Of the flies released in forest edge, approximately 16% of the recaptured females were caught in Shannon traps in the peridomicile habitat. The results indicate that the movements of L. neivai are spatially focal and the possibility of dispersion from forest to peridomicile habitat may be an important way of contracting leishmaniasis in dwellings.     

Bionomics of Phlebotomus papatasi (Diptera: Psychodidae) in an endemic focus of zoonotic cutaneous leishmaniasis in central Iran.

Following an epidemiological survey of zoonotic cutaneous leishmaniasis (ZCL) in several villages of Badrood, a rural district north of the city of Natanz, central Iran, Phlebotomus (Phlebotomus) papatasi Scopoli were found to be naturally infected with Leishmania (Leishmania) major zymodeme MON-26. Sand flies were collected and dissected biweekly from rodent burrows from May to October 2001. Leptomonad infection rates varied between 6.7% and 22.0%, being greatest in September, coinciding with peak activity of P. papatasi, two-three months before the highest incidence of ZCL human cases in November-December. The leptomonad infection rate was 1.1% of the 94 P. papatasi captured indoors. In ELISA testing of 520 P. papatasi blood meals during Sept. 2001 and Aug. 2002, the proportion giving positive reactions for human, sheep, cow, goat, rodent, and bird were 31.2%, 69.6%, 63%, 38.8%, 24.7%, and 21.8%, respectively. This report thus incriminates P. papatasi as the vector of zoonotic cutaneous leishmaniasis in this part of Iran.     

Sand fly (Diptera: Psychodidae) distribution in the endemic and non‐endemic foci of visceral leishmaniasis in northwestern Iran

An entomological study was conducted from June to September, 2010 in rural regions of Azarbayjan‐e‐sharqi, Azarbayjan‐e‐qarbi, and Ardabil provinces in northwestern Iran to determine sand fly fauna, diversity, and distribution in different habitats and altitudes using both sticky papers and light traps. Geographical distribution of sand flies and the similarity of populations in different locations were analyzed ecologically based on the Shannon‐Wiener Index and Jacard Coefficient, respectively. A total of 3,982 specimens was collected and sixteen species recorded. They belonged to the genera Phlebotomus [subgenus Phlebotomus (P. papatasi), Paraphlebotomus (P. sergenti, P. mongolensis, P. caucasicus, P. jacusieli), Larroussius (P. major s.l., P. tobbi, P. perfiliewi transcaucasicus, P. kandelakii) and Adlerius (P. halepensis, P. brevis, P. longiductus, P. balcanicus)], and Sergentomyia [subgenus Sergentomyia (S. sintoni, S. dentata and S. theodori)]. P. papatasi was the predominant species in all the locations except Bileh Savar, Macu, and Meshkin Shahr, followed by P. perfiliewi transcaucasicus and P. kandelakii. The latter species were caught from different habitats and altitudes with higher frequency than other species of the subgenus Larroussius. The lowest abundance belonged to P. jacusieli. The predominant species of subgenus Adlerius was P. halepensis. Data analysis showed that Meshkin Shahr and Bileh Savar had high and low diversities of sand fly distribution, respectively. Meshkin Shahr and Sarab districts had the highest similarity. Both are located in the foothills of Sabalan Mountain, with high diversity and richness.     

Identification of phlebotomine sand flies (Diptera: Psychodidae) from leishmaniasis endemic areas in southeastern Mexico using DNA barcoding

Leishmaniasis, a vector‐borne disease transmitted to humans through the bite of phlebotomine sand flies, is of public health significance in southeastern Mexico. Active and continuous monitoring of vectors is an important aspect of disease control for the prediction of potential outbreaks. Thus, the correct identification of vectors is paramount in this regard. In this study, we employed DNA barcoding as a tool for identifying phlebotomine sand flies collected in localized cutaneous leishmaniasis endemic areas of Quintana Roo, Mexico. Specimens were collected using CDC light and Shannon traps as part of the Mexican Ministry of Health surveillance program. DNA extraction was carried out using a nondestructive protocol, and morphological identification based on taxonomic keys was conducted on slide‐mounted specimens. Molecular taxonomic resolution using the 658‐bp fragment of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene was 100% congruent with the morphological identification. Seven species were identified: Lutzomyia cruciata (Coquillett 1907), Lutzomyia longipalpis (Lutz & Neiva 1912), Psathyromyia shannoni (Dyar 1929), Dampfomyia deleoni (Fairchild & Hertig 1947), Dampfomyia beltrani/steatopyga (Vargas & Díaz‐Nájera 1951), Bichromomyia olmeca olmeca (Vargas & Díaz‐Nájera, 1959), and Brumptomyia mesai (Sherlock 1962). Mean intraspecific divergence ranged from 0.12% to 1.22%, while interspecific distances ranged from 11.59% to 19.29%. Neighbor‐joining (NJ) analysis using the Kimura 2‐parameter model also showed specimens of the same species to be clustered together. The study provides the first cox1 sequences for three species of sand flies and indicates the utility of DNA barcoding for phlebotomine sand flies species identification in southeastern Mexico.     

Detection of Leishmania amazonensis and Leishmania braziliensis in Culicoides (Diptera,Ceratopogonidae) in an endemic area of cutaneous leishmaniasis in the Brazilian Amazonia

Biting midges in the genus Culicoides act as vectors of arboviruses throughout the world and as vectors of filariasis in Latin America, the Caribbean, and parts of Africa. Although Culicoides spp. are currently not considered to be vectors of Leishmania protozoa, the high abundance of biting midges in areas with active cutaneous leishmaniasis transmission points to the possibility of Culicoides infection by these pathogens. We used PCR to test captured Culicoides species for natural infection with Leishmania spp. We tested 450 Culicoides females, divided into 30 pools of 15 individuals each, as follows: nine pools of C. foxi (135 specimens), seven pools of C. filariferus (105), seven pools of C. insignis (105), five pools of C. ignacioi (75), and two pools of C. flavivenula (30). PCR confirmed the presence of Leishmania braziliensis DNA in C. ignacioi (0.14%), C. insignis (0.14%), and C. foxi (0.11); and Le. amazonensis DNA in C. filariferus (0.14%) and C. flavivenula (0.50%). We conclude that these Culicoides species can be naturally infected, but vector competence and transmission capability must be confirmed in future studies. Our results warrant further investigation into the role of these biting midge species in the leishmaniasis epidemiological cycle.     

Vertical stratification and development aspects of phlebotomine sand flies (Diptera: Psychodidae) in an area of Atlantic Forest tree species in a metropolitan region in northeastern Brazil.

In the state of Rio Grande do Norte in northeast Brazil, cases of visceral leishmaniasis (VL) occur mainly in the periurban areas of the city of Natal. Lutzomyia longipalpis Lutz & Neiva 1912 (Diptera: Psychodidae), a vector of Leishmania chagasi (Protozoa: Trypanosomatidae) to humans, is found throughout the state. Flora and fauna influence the distribution of sand fly species, whose horizontal or vertical stratification can be used as a parameter for identifying potential vectors, considering the presence of vertebrate hosts in the area. The purpose of this study was to obtain information about the vertical stratification of phlebotomine sand flies in an endemic area of leishmaniasis in Rio Grande do Norte, and associate it with the presence of other animals in the peridomiciliary environment as well as to analyze, under laboratory conditions, aspects of L. longipalpis reproduction in wild females. The sand flies were captured with light traps hung at different heights in species of Atlantic Forest trees and in a peridomiciliary environment in animal shelters. The traps were placed between 17:30 and 6:00 of the following day, in a peridomiciliary and extradomiciliary area of a forest fragment in both dry and rainy months. In the extradomiciliary environment, the traps were installed at 1, 3 and 5 m above the ground. The biological cycle of L. longipalpis was followed from the eggs of 200 wild females. Specimens of L. lenti, L. walkeri, and L. migonei were captured. The comparison and statistical analysis showed that L. longipalpis is more abundant at a height of 3 m and L. evandroi at 1 m. In the animal shelters (chickens, horses, and armadillos), we captured mainly specimens of L. longipalpis and L. evandroi. The duration of the biological cycle of L. longipalpis was approximately 38 days at a temperature of 28 degrees C.     

Sandflies (Diptera: Psychodidae) in a focus of visceral leishmaniasis in White Nile, Sudan

Visceral leishmaniasis (VL) has been known to occur since the 1980s on the western bank of the White Nile River (Central Sudan), 150 km south of Khartoum, and has resulted in high mortality. The most recent outbreak of the disease in this area began in 2006. Entomological surveys were carried out during May 2008, June 2010 and May and July 2011 in the White Nile area. Sandflies were collected using Centers for Disease Control light traps and sticky oil traps in the village of Kadaba and the nearby woodland. Phlebotomus females were dissected for the presence of Leishmania promastigotes. A total of 17,387 sandflies, including six species of Phlebotomus and 10 species of Sergentomyia, were identified. The Phlebotomus species recorded were Phlebotomus orientalis, Phlebotomus papatasi, Phlebotomus bergeroti, Phlebotomus duboscqi, Phlebotomus rodhaini and Phlebotomus saevus. P. orientalis was collected in both habitats. The relative abundance of P. orientalis in the woodland habitat was higher than that recorded in the village habitat. In the woodland habitat, there was a notable increase in the relative abundance of P. orientalis during the surveys conducted in 2008 and 2010 compared to 2011. None of the 311 P. orientalis females dissected were infected with Leishmania promastigotes, although relatively high parous rates were recorded in both habitats. Based on the distribution of P. orientalis recorded in this study, this species is the most likely vector of VL in the endemic focus in the White Nile area. Further investigation is required to elucidate the seasonal abundance and distribution of the vector, as well as the transmission season of VL in both habitats so that appropriate control strategies for the vector can be designed.     

Ecological interactions among phlebotomines (Diptera: Psychodidae) in an agroforestry environment of northeast Brazil

Phlebotomine vectors transmit parasites and can cause visceral leishmaniasis (VL) or cutaneous leishmaniasis (TL). Phlebotomine females are hematophagous but need to ingest carbohydrates, possibly promoting the development of protozoan parasites in their digestive tract. The present study evaluated the species composition and abundance across several habitats in a metropolitan landscape, as well as associations among phlebotomines, plants, and local climatic parameters. Three consecutive monthly collections were carried out in an Atlantic Forest fragment, using CDC light traps in peridomestic areas and cashew, coconut, and mango tree. plantations. Eight species of phlebotomine were captured: Evandromyia evandroi, Lutzomyia longipalpis, Psathyromyia shannoni, Sciopemyia sordellii, Evandromyia walkeri, Psychodopygus wellcomei, Nyssomyia whitmani, and Nyssomyia intermedia, primarily from the forest environment. L. longipalpis was confirmed as a species adapted to anthropic environments, while P. wellcomei was shown to be predominately forest‐dwelling. Phlebotomines exhibited diversified food consumption patterns in relation to carbohydrate sources. They fed on both native and exotic species of arboreal and shrubby vegetables and gramineous plants.     

Distribution of sand flies (Diptera: Psychodidae) at different altitudes in an endemic region of American cutaneous leishmaniasis in the State of Espírito Santo, Brazil.

The involvement of different sand fly species in the transmission of American cutaneous leishmaniasis (ACL) at different altitudes was evaluated in the municipality of Afonso Cláudio in the State of Espírito Santo, Brazil, from November 1995 to February 1997. CDC light traps and Shannon traps baited with human volunteers were hung simultaneously at three altitudes of a river valley: (a) 650-750 m in an area of active ACL transmission; (b) 750-850 m in a transitional area; and (c) 850-950 m in an area where no ACL transmission occurred. A total of 13,363 specimens belonging to 28 species was collected. The five most abundant man-biting species were Lutzomyia intermedia, which constituted 24.3% of the total, Lu. migonei (22.3%), Lu. whitmani (15.4%), Lu. fischeri (14.9%) and Lu. monticola (5.8%). Analysis of the distribution of these species at the three altitudes provided evidence that Lu. fischeri and Lu. monticola were not involved in ACL transmission, whereas Lu. migonei and Lu. whitmani might act as secondary vectors of Leishmania (Viannia) braziliensis and Lu. intermedia was probably the principal vector of the parasite. These results reinforce those published in the existing literature, which indicate that Lu. intermedia is the main vector of Le. (V.) braziliensis in Southeast Brazil, while Lu. migonei and Lu. whitmani are of secondary importance.     

Molecular detection of Leishmania tropica in field caught Phlebotomus sand flies (Diptera: Psychodidae) from cutaneous leishmaniasis endemic focus of Pakistan

Cutaneous Leishmaniasis is endemic in tribal district Khyber for last more than one decade. The causative agent Leishmania tropica is known but sand fly species responsible for the transmission of disease still needs to be investigated. A total of 2647 Phlebotomus females belonging to 11 species were divided into 435 batches and subjected to PCR for detection of Leishmania in sand flies. A total of 50 batches belonging to three species i.e. Phlebotomus sergenti, Phlebotomus papatasi and Phlebotomus alexandri were detected positive for Leishmania tropica. Overall minimum infection rate was 1.89% (50/2647). Highest minimum infection rate of 2.11% (39/1710) was observed for Phlebotomus sergenti followed by 1.21% (8/661) for Phlebotomus paptasi and 1.82% (3/165) for Phlebotomus alexandri. Both blood fed (38%) and unfed (62%) sand flies were detected positive for the parasite DNA. Positive specimens were collected throughout the active season, from all collection sites of the study area. Detection of Leishmania parasite in multiple species of Phlebotomus indicates the possible role of these species as vector of disease in the tribal district Khyber of Pakistan. It also indicates the probable complex transmission cycle of the disease involving multiple vector species in the study area. Devising a control strategy by focusing on these vector species, may reduce the disease burden in the cutaneous leishmaniasis endemic tribal district Khyber.     

The sandfly fauna,anthropophily and the seasonal activities of Pintomyia spinicrassa (Diptera: Psychodidae: Phlebotominae) in a focus of cutaneous leishmaniasis in northeastern Colombia

This study was conducted to identify the sandfly fauna and the anthropophilic species in a coffee-growing area of Villanueva, Norte de Santander, Colombia, a focus of American cutaneous leishmaniasis, and to analyse the relationship between the most frequent species and rainfall, relative humidity and temperature, with the aim of contributing to epidemiological surveillance in the area. Sandfly collections were performed fortnightly between February 2006-September 2007 using automatic light traps, Shannon traps, protected human bait and aspiration in resting places. A total of 7,051 sandflies belonging to 12 species were captured. Pintomyia spinicrassa (95.7%) predominated. Pintomyia oresbia and Lutzomyia sp. of Pichinde were found in the state of Norte de Santander for the first time. Pi. spinicrassa, Pintomyia nuneztovari, Micropygomyia venezuelensis, Lutzomyia (Helcocyrtomyia) scorzai and Lu. (Helcocyrtomyia) sp. were captured on the protected human bait. A significant association between Pi. spinicrassa abundance and the total rainfall and the average temperature and humidity 10 days before the collection was observed. The dominance of Pi. spinicrassa, a recognised vector of Leishmania braziliensis, especially during the dry periods, indicates that the risk of parasite transmission may increase.     

Carbohydrate digestion in Lutzomyia longipalpis' larvae (Diptera - Psychodidae)

Lutzomyia longipalpis is the principal species of phlebotomine incriminated as vector of Leishmania infantum, the etiological agent of visceral leishmaniasis in the Americas. Despite its importance as vector, almost nothing related to the larval biology, especially about its digestive system has been published. The objective of the present study was to obtain an overview of carbohydrate digestion by the larvae. Taking in account that phlebotomine larvae live in the soil rich in decaying materials and microorganisms we searched principally for enzymes capable to hydrolyze carbohydrates present in this kind of substrate. The principal carbohydrases encountered in the midgut were partially characterized. One of them is a α-amylase present in the anterior midgut. It is probably involved with the digestion of glycogen, the reserve carbohydrate of fungi. Two other especially active enzymes were present in the posterior midgut, a membrane bound α-glucosidase and a membrane bound trehalase. The first, complete the digestion of glycogen and the other probably acts in the digestion of trehalose, a carbohydrate usually encountered in microorganisms undergoing hydric stress. In a screening done with the use of p-nitrophenyl-derived substrates other less active enzymes were also observed in the midgut. A general view of carbohydrate digestion in L. longipalpis was presented. Our results indicate that soil microorganisms appear to be the main source of nutrients for the larvae.     

Identification of the natural breeding sites of sandflies (Diptera: Psychodidae: Phlebotominae), potential vectors of leishmaniasis, in the province of Chaco, Argentina

The aim of this work was to identify the natural breeding sites of sandflies in the province of Chaco, Argentina, for the first time. Preliminary studies were conducted in two different phytogeographic regions: dry Chaco (Parque Provincial Pampa del Indio), in January 2010, and humid Chaco (Resistencia, Margarita Belén and Colonia Benítez), from May-September 2010. A total of 127 samples were collected (Pampa del Indio: 15, Resistencia: 37, Margarita Belén: 36, Colonia Benítez: 39). A female of Migonemyia migonei was found in Pampa del Indio at the base of a bromeliad in the summer (January) and a pupal exuvium of a phlebotomine fly was found in Resistencia, in a place where dogs rested, in the winter (July). These findings highlighted these two sites as potential breeding sites. Because the existence of potential natural breeding sites for sandflies has been demonstrated in both forest and periurban areas, expanding the search efforts and characterising these sites will enable the development of specific study designs to gain insight into the spatial distribution of the risks posed by these vectors. The resulting information will serve as a basis for proposing and evaluating vector control measures.     

Phylogenetic relationships in the subfamily Psychodinae (Diptera,Psychodidae)

Espíndola, A., Buerki, S., Jacquier, A., Je?ek, J. & Alvarez, N. (2012). Phylogenetic relationships in the subfamily Psychodinae (Diptera, Psychodidae). —Zoologica Scripta, 41, 489–498. Thanks to recent advances in molecular systematics, our knowledge of phylogenetic relationships within the order Diptera has dramatically improved. However, relationships at lower taxonomic levels remain poorly investigated in several neglected groups, such as the highly diversified moth‐fly subfamily Psychodinae (Lower Diptera), which occurs in numerous terrestrial ecosystems. In this study, we aimed to understand the phylogenetic relationships among 52 Palearctic taxa from all currently known Palearctic tribes and subtribes of this subfamily, based on mitochondrial DNA. Our results demonstrate that in light of the classical systematics of Psychodinae, none of the tribes sensu Je?ek or sensu Vaillant is monophyletic, whereas at least five of the 12 sampled genera were not monophyletic. The results presented in this study provide a valuable backbone for future work aiming at identifying morphological synapomorphies to propose a new tribal classification.