Sand fly species of Sanliurfa province in Turkey

The species composition and seasonal abundance of sand flies (Diptera: Phlebotominae) were studied in the years 2000-2002 in the Sanliurfa region, which is the largest focus of cutaneous leishmaniasis in south-eastern Turkey. Sixteen species were identified among 29 771 specimens collected at 17 different sites by light traps, sticky papers and aspirators. The most common species were Phlebotomus papatasi (Scopoli) (45.4%), P. perfiliewi Parrot (21.9%), and P. sergenti Parrot (19.4%). The other species found were P. major Adler & Theodor (3%), P. neglectus Leger & Pesson (2.2%), P. brevis Theodor & Mesghali (2%), P. alexandri Sinton (1.9%), P. galilaeus Theodor (1.6%), P. halepensis Theodor (0.84%), Sergentomyia adleri Theodor (0.78%), S. dentata Sinton (0.49%), S. minuta Rondani (0.42%), S. theodori Parrot (0.16%), P. kazeruni Theodor & Mesghali (0.001%) and P. mascitti Grassi (0.001%) and one unidentified Phlebotomus species. Among these species P. galilaeus, S. minuta and S. dentata are the first records for this area. All species showed seasonal fluctuations, with the period of highest abundance between May and October.     

Recent findings of Phlebotomus neglectus Tonnoir, 1921 in Italy and its western limit of distribution

During the sand fly season of 2001, sand flies were collected in two different regions of Italy to investigate the northern distribution of Phlebotomus neglectus. The study areas were two provinces of northern Italy, Ivrea and Verona, in Piedmont and Veneto regions respectively. Sticky traps and CDC miniature light traps were used both inside and outside domestic and peri-domestic environments. Sand flies were also collected in domestic resting sites with mouth and electric aspirators. Of 1,295 sand fly specimens collected, 855 were from the Veneto region and 440 were from the Piedmont region. Five species of the genera Phlebotomus and Sergentomyia were identified. S. minuta (66.3%) was the most abundant sand fly followed by P. perniciosus (22.2%), P. neglectus (10.3%) and P. papatasi (1.2%). Only one specimen of P. mascittii was caught in Piedmont region. P. neglectus was found in 9 out of 15 (60%) of the sites where sand flies were found with a prevalence of 1.5% to 45.8%; the highest value was recorded in Piedmont region. P. neglectus was mostly associated with peridomestic and domestic sites. The results confirm previous observations on the presence of P. neglectus in northern Italy and suggest that this species is probably widespread in the area. From a review of all previous published and unpublished records of P. neglectus, it appears that this species is present in only two areas of the country, namely in the regions of Friuli Venezia Giulia, Veneto and Piedmont in northern Italy and the regions of Calabria, Apulia and Sicily in southern Italy. The apparent absence of P. neglectus in Central Italy is discussed and the resulting settlement through two probable migration routes from the East is analysed.     

Distribution and altitudinal structuring of phlebotomine sand flies (Diptera: Psychodidae) in southern Anatolia, Turkey: their relation to human cutaneous leishmaniasis.

The two Old World genera, Phlebotomus and Sergentomyia, were both recorded in southern Anatolia in Turkey. Phlebotomus species predominated and comprised about 93% of the entire collection (3,172 specimens). Out of the sixteen species identified, two belonged to the genus Sergentomyia: S. dentata and S. theodori. The remaining fourteen species in the genus Phlebotomus were grouped under four subgenera including some species that are elsewhere known to act as vectors of human cutaneous leishmaniasis. Most of the Phlebotomus were P. tobbi (32.5%), but P. papatasi, P. transcaucasicus, P. halepensis, P. galilaeus, P. sergenti, P. syriacus, P. neglectus, P. simici, P. alexandri, P. similis, P jacusieli, P. perfiliewi, and P. brevis were also identified. There were two associations of sand fly fauna with altitudinal gradient; the first one at relatively higher altitudes and the second one at lower altitudes. The transition between these two assemblages was within the range of 800-1,000 m. It is likely that Adana and Hatay provinces are transitional areas between western and eastern Anatolia. Mountains do not appear to be important geographical barriers for sand fly distribution. We also found that the proven vector P. sergenti is a widely distributed species throughout southern Anatolia and this species, together with its closely related species P. similis, shows sympatry in Konya Province.     

Distribution of phlebotomine sandflies and the rate of their infection with Leishmania promastigotes in the Algarve, Portugal

Four species of sandflies: Phlebotomus (Larroussius) perniciosus Newstead, Sergentomyia minuta (Rondani), Phlebotomus (Paraphlebotomus) sergenti Parrot and Phlebotomus (Larroussius) ariasi Tonnoir, were collected, by aspiration and light traps, from three dog kennels and an area of high prevalence of human and canine visceral leishmaniasis in the Algarve, Portugal. Midgut infections with leishmanial promastigotes were observed only in P. perniciosus with a minimum infection rate of 5%. The engorgement, gravidity, population dynamics and sex ratios of the sandflies, and the temperature, source and altitude of collection sites were examined. Host preferences were noted for P. perniciosus.     

Cutaneous leishmaniasis in western Sicily (Italy) and preliminary survey of phlebotomine sandflies (Diptera: Psychodidae)

A survey of phlebotomine sandflies was carried out in two provinces of western Sicily (Italy), where 65 cases of cutaneous leishmaniasis (CL) had been diagnosed by the Dermatology Department of the University of Palermo, between 1977 and 1987. Eight collecting stations, distributed throughout the CL foci of the Agrigento and Palermo provinces produced a total of 2,410 specimens (12.78% males). Of these, 77.59% were Phlebotomus perfiliewi, 12.78% P. perniciosus, 0.74% P. major and 2.07% Sergentomyia minuta. P. perfiliewi, the probable vector of CL, was present in 7 out of the 8 collecting stations with very high densities in two localities.     

Genetic polymorphism in sympatric species of the genus Phlebotomus, with special reference to Phlebotomus perniciosus and Phlebotomus longicuspis (Diptera, Phlebotomidae)

The Random Amplified Polymorphic DNA assay was used to study genetic variation within and between five Phlebotomus species belonging to three subgenera: P. (Larroussius) ariasi, P. (L.) longicuspis, P. (L.) perniciosus, P.(Paraphlebotomus) sergenti and P. (Phlebotomus) papatasi sympatric in southern Spain and proven vector of leishmaniasis. Two cluster analysis were proposed: one according to sandfly species and populations, the second according individual specimens of Phlebotomus perniciosus, Phlebotomus longicuspis s.l. and intermediate morphological specimens between these species. The results obtained are closely correlated with the taxonomy classically accepted for the subgenera and with the automatic classifications made by other authors which use morphological and isoenzymatic data. The validity of the species Phlebotomus longicuspis is also discussed.     

Limits of a rapid identification of common Mediterranean sandflies using polymerase chain reaction-restriction fragment length polymorphism

A total of 131 phlebotomine Algerian sandflies have been processed in the present study. They belong to the species Phlebotomus bergeroti, Phlebotomus alexandri, Phlebotomus sergenti, Phlebotomus chabaudi, Phlebotomus riouxi, Phlebotomus perniciosus, Phlebotomus longicuspis, Phlebotomus perfiliewi, Phlebotomus ariasi, Phlebotomus chadlii, Sergentomyia fallax, Sergentomyia minuta, Sergentomyia antennata, Sergentomyia schwetzi, Sergentomyia clydei, Sergentomyia christophersi and Grassomyia dreyfussi. They have been characterised by sequencing of a part of the cytochrome b (cyt b), t RNA serine and NADH1 on the one hand and of the cytochrome C oxidase I of the mitochondrial DNA (mtDNA) on the other hand. Our study highlights two sympatric populations within P. sergenti in the area of its type-locality and new haplotypes of P. perniciosus and P. longicuspis without recording the specimens called lcx previously found in North Africa. We tried to use a polymerase chain reaction-restriction fragment length polymorphism method based on a combined double digestion of each marker. These method is not interesting to identify sandflies all over the Mediterranean Basin.     

Entomological and parasitological study on phlebotomine sandflies in central and northern Albania

An entomological survey was carried out in two districts of central (Kruje) and northern (Lezhe) Albania. Six collecting sites, showing a variety of diurnal resting sites, were monitored for adult sandflies from June through October 2002. Flies were collected with CDC miniature light traps, sticky traps and mechanical or hand aspirators in peridomestic sites, in bedrooms and inside cow barns, chicken coops and pigpens. All collecting sites monitored were found positive for sandflies. A total of 849 specimens were caught (29.2% males) belonging to five Phlebotomus species. Phlebotomus neglectus (75.6%) was the most abundant species followed by P. perfiliewi (14.4%), P. papatasi (4.6%), P. tobbi (3.6%) and P. similis (1.8%). The first adult of P. neglectus appeared on June 11 and the last one was collected on October 16. The highest density for this species was observed at the end of July. A total of 111 blood-fed females were caught from the two areas studied. P. neglectus was the only species found blood fed in Lezhe and the same species was prevalent (56.1%) in Kruje followed by P. perfiliewi (30.3%), P. tobbi (10.6%); P. papatasi was represented by only two specimens. Blood meal origin was determined in 45/66 (68.2%) of the females tested from Kruie district. P. neglectus was found fed on four hosts, showing the following feeding patterns: cow (71.4%), dog (117.1%), chicken (5.7%) and human (5.7%); P. perfiliewi was found fed on cow (80.0%) and chicken (20.0%), P. tobbi on cow (50.0%), chicken (25.0%) and dog (25.0%). One specimen of P. papatasi was found fed on cow. When such prevalences were analysed by the available biomass for each host present at the collecting site, P. neglectus resulted to be an opportunistic feeder rather than exhibiting preferences for any specific animal. PCR analysis of 39 P. neglectus from the Lezhe district gave negative results for the presence of Leishmania DNA.     

Bionomics of phlebotomine sand flies (Diptera: Psychodidae) in the province of Al-Baha, Saudi Arabia

The bionomics of phlebotomine sand flies (Diptera: Psychodidae) were studied for two successive years (January 1996-December 1997) at 12 collecting stations representing six sectors of the province of Al-Baha, Saudi Arabia. The predominant species was Phlebotomus bergeroti (41.7%), followed by lesser numbers of Phlebotomus sergenti (11%), Phlebotomus arabicus (10.6%), Sergentomyia tiberiadis (10.5%), Phlebotomus papatasi (10.2%), Sergentomyia antennata (9.6%), Phlebotomus alexandri (3%), Phlebotomus orientalis (2.3%) and Sergentomyia clydei (1.1%). The distribution of the collected species including species that are elsewhere known to act as vectors of human cutaneous leishmaniasis were distributed across different altitudes in Al-Baha. P. bergeroti, P. papatasi and P. arabicus were more abundant indoors; however, P. sergenti was more abundant outdoors. Sand fly populations exhibited three patterns of seasonal abundance in terms of their monthly activity. P. bergeroti, P. sergenti and P. arabicus were found to be naturally infected with Leishmania-like flagellates at an infection rate of 0.2%.     

Phlebotomine sandflies (Diptera-Psychodidae) of the isle of Cyprus. II--Isolation and typing of Leishmania (Leishmania infantum Nicolle, 1908 (zymodeme MON 1) from Phlebotomus (Larroussius) tobbi Adler and Theodor, 1930

During two surveys conducted in Cyprus (August 1998 and September 1999), 2,910 phlebotomine sandflies females were caught by CDC miniature light traps then dissected under binocular and examined on microscope. Eleven species were identified: Phlebotomus papatasi, P. sergenti, P. jacusieli, P. alexandri, P. tobbi, P. galilaeus, P. mascittii, P. economidesi, Sergentomyia fallax, S. minuta et S. azizi. The Larroussius species (P. galilaeus and P. tobbi) are the most abundant (more than 60% of our captures). Promastigotes were isolated from one specimen identified as P. tobbi. A Leishmania stock was successfully cultured and identified by isoenzyme characterisation as belonging to L. infantum zymodeme MON 1. The same zymodeme was isolated and identified from four dogs too. Because of the absence of usual vectors of L. infantum in the eastern part of the Mediterranean basin (P. neglectus and P. syriacus), and according to its distribution in Cyprus, P. tobbi constitute certainly a good local vector. It seems to be not very anthropophilic, that could explain the very few human cases.     

Entomological survey of sandflies (Diptera, Psychodidae) in the province of L'Aquila (Abruzzo)]

A survey on phlebotomine sandflies (Diptera, Psychodidae) was carried out in the province of L'Aquila, Abruzzo region (Italy). Ten collecting sites at altitudes between 700 and 1,070 m a.s.l., produced a total of 324 specimens (81.1% males), 72.86% being Phlebotomus perniciosus, 3.08% P. mascittii, 0.30% P. papatasi and 23.76% Sergentomyia minuta. The collecting sites at 1,070, 950 and 775 m a.s.l. are the highest levels recorded in Italy for P. perniciosus, P. mascittii and P. papatasi, respectively. P. perfiliewi, the proven vector of cutaneous leishmaniasis (CL) in the Abruzzo region (Maroli et al., 1987) was absent. The probable role of P. perniciosus in the transmission of the sporadic CL cases reported from the area is discussed.     

Phlebotominae (Diptera: Psychodidae) of human leishmaniosis sites in Tunisia

In order to identify the phlebotomine sandfly populations in Tunisian leishmaniosis foci, an entomological survey was carried out through three entomological seasons (2002-2003-2004) in 19 visceral and cutaneous leishmaniosis areas, located in six bioclimatic zones. Sandfly collections were based on light and sticky traps placed around human leishmaniosis cases. 8,722 phlebotomine sandflies belonging to 12 species were collected. The dominance of subgenus Larroussius species in northern foci, Phlebotomus papatasi in south-western foci and their co-dominance in the centre of the country is in accordance with the distribution of Leishmania infantum and L. major in Tunisia. The low density found in the historical zoonotic cutaneous leishmaniosis focus of Metlaoui in the south-west may indicate the high competence of the local populations. Studied phlebotomine settlements have showed a low specific diversity in most of the studied sites. In L. infantum areas, the dominant species were respectively: P. perfiliewi in the cutaneous leishmaniosis site of the humid bioclimatic stage, P. perniciosus in the cutaneous and visceral leishmaniosis foci of semi-arid and arid bioclimatic stages and P. longicuspis in the visceral leishmaniosis focus of saharan bioclimate. In the zoonotic cutaneous leishmaniosis foci, P. papatasi was a dominant species. In the well-known south-eastern foci of cutaneous leishmaniosis due to L. killicki, P. sergenti was a dominant species with P. perniciosus. In the central emerging foci of L. killicki, P. perniciosus was a dominant species in some sites whereas it was very rare in others. In these sites, the subgenus Paraphlebotomus was always present with a higher abundance of P. alexandri than P. sergenti.     

Phlebotomines of the Isle of Cyprus. III. Species inventory

Three surveys on canine leishmaniasis were carried out in Cyprus (1993, 1998 and 1999) emphasise the presence of eleven species of phlebotomine sandflies: Phlebotomus (Phlebotomus) papatasi, P. (Paraphlebotomus) alexandri, P. (Pa.) jacusieli (first mention in Cyprus), P. (Pa.) sergenti, P. (Larroussius) galilaeus, P. (L.) tobbi, P. (Transphlebotomus) economidesi, P. (T.) mascittii, Sergentomyia (Sergentomyia) azizi (its specific statute is validated by the authors), S. (S.) fallax et S. (S.) minuta. P. (Adlerius) kyreniae was not caught during the surveys. The authors propose hypothesis of settlement of the island by phlebotomine sandflies according this species inventory. A first migration period took probably place during the Miocene time and a second one during the Pleistocene time.     

Bionomics of phlebotomine sandflies in the Galilee focus of cutaneous leishmaniasis in northern Israel

The bionomics of phlebotomine sandflies (Diptera: Psychodidae) were studied for three years (2001-2003) in the Galilee focus of cutaneous leishmaniasis in northern Israel, where the causative Leishmania tropica (Kinetoplastida: Trypanosomatidae) is transmitted by Phlebotomus (Adlerius) arabicus Theodor and Phlebotomus (Paraphlebotomus) sergenti Parrot, comprising 22% and 8%, respectively, of the local sandfly fauna sampled by light traps. The predominant species overall was Phlebotomus (Larroussius) tobbi Adler & Theodor (51%) with lesser numbers of Phlebotomus (Adlerius) simici Theodor (11%), Phlebotomus (Larroussius) syriacus Adler & Theodor (5%), Phlebotomus (Larroussius) perfiliewi Perfil'ev (3%) and Phlebotomus (Phlebotomus) papatasi Scopoli (0.05%). Sandfly adult populations were prevalent from April to November and peaked between June and August, being more abundant through the summer in irrigated habitats, such as gardens and orchards, than in open grassland. Of the two cutaneous leishmaniasis vectors, P. sergenti preferred boulder mounds located at the outskirts of settlements, whereas P. arabicus was more abundant overall and near houses in particular. Females of all these sandfly species displayed a peak of activity after sunset (20.00-22.00 hours), whereas activity of males persisted longer through the night. Another slight increase in activity was noted before dawn (02.00-04.00 hours). Phlebotomus arabicus appears to be the main vector of L. tropica in the Galilee focus, due to its denser populations, more endophily and preference for peridomestic habitats than shown by P. sergenti in northern Israel.     

Aspects of the bionomics of Phlebotomus sergenti sandflies from an endemic area of anthroponotic cutaneous leishmaniasis in Aleppo Governorate, Syria

Aspects of the bionomics of phlebotomine sandflies (Diptera: Psychodidae) were studied from June to November 2005 in three foci of anthroponotic cutaneous leishmaniasis (ACL) in Aleppo Governorate, Syria, where the agent Leishmania tropica (Wright) (Kinetoplastida: Trypanosomatidae) is transmitted by Phlebotomus sergenti Parrot. Syria has been designated by the World Health Organization as one of four countries in the Old World where cutaneous leishmaniasis is hyperendemic, but little is known about the biology of local vector populations. Standard collections by sticky traps showed two peaks in density, in June and late August. In total, 1840 sandflies were caught, comprising five species: Phlebotomus papatasi (Scopoli) (68.0%); P. sergenti (25.4%); Sergentomyia minuta (Rondani) (6.4%); Phlebotomus tobbi Adler & Theodor (0.1%), and Phlebotomus mascittii canaaniticus Adler & Theodor (0.1%). Similar numbers of P. sergenti were caught indoors (246 specimens) and outdoors (222), whereas P. papatasi was significantly more abundant indoors (1096 specimens) than outdoors (156) (χ² = 241, P < 0.01). In total, 212 blood-fed females were tested for host blood determination, of which 176 (83.0%) reacted with anti-species reagent. Results from 20 P. sergenti suggest that this species is an opportunistic feeder, imbibing human, ovine, avian, bovine and feline blood, although more bloodmeals were taken from humans and cattle than expected in relation to the relative proportions of potential hosts present (the forage ratio, FR). The bionomics of P. sergenti are discussed in relation to the inefficacy of control campaigns based on indoor spraying with residual insecticides that have been implemented by the Syrian Ministry of Health to control the epidemics of ACL in the Aleppo Governorate.     

Phlebotomus sergenti (Parrot, 1917) identified as Leishmania killicki host in Ghardaïa, south Algeria

Since 2005, an outbreak of human cutaneous leishmaniasis (CL) in Gharda?a, south Algeria, was studied and one output of these investigations was the identification of two Leishmania species, Leishmania major and Leishmania killicki, as the CL causative agents. In the present study, we were curious to focus on sand fly fauna present in this area and detection of Leishmania-positive sand fly females. Sand flies (3717) were collected during two seasons using sticky papers and CDC light traps in urban, rural and sylvatic sites. Twelve Phlebotomus species were identified. Phlebotomus papatasi was dominant in the urban site while Phlebotomus sergenti and Phlebotomus riouxi/chabaudi were dominant in the sylvatic site. Out of 74 P. sergenti females captured by CDC light traps in the sylvatic site populated by Gharda?as' Gundi (Massoutiera mzabi), three ones were hosting Leishmania promastigotes. PCR-RFLP and sequencing of seven single-copy coding DNA sequences identified the promastigotes as L. killicki. Furthermore, laboratory experiments revealed that L. killicki isolate sampled from a CL patient inhabiting the studied region develop well in P. sergenti females. Our findings strongly suggest that the human cutaneous leishmaniases caused by L. killicki is a zoonotic disease with P. sergenti sand flies acting as hosts and vectors and gundi rodents as reservoirs.     

Ootaxonomy and eggshell ultrastructure of Phlebotomus sandflies

The eggshell structure of four sandfly species: Phlebotomus perniciosus Newstead, P.perfiliewi Parrot, P.papatasi Scopoli and P.duboscqi Neveu-Lemaire, was examined by scanning and transmission electron microscopy (SEM and TEM). At the TEM level, the eggshell appears to have a homogeneous vitelline envelope and a thick chorion. At SEM level, the eggshell of all species is characterized by the outer chorion forming a series of fifteen to twenty longitudinal sinuous ridges, cross-linked in places to form a pattern of polygons, each line of the chorion consisting of columns arranged in a palisade. The aeropyle region of the egg is described for the first time in phlebotomine sandflies. Specific characters of the eggshell topography are described for distinguishing between these and other species of Phlebotomus.     

Leishmaniasis in Sardinia. III. Soil analysis of a breeding site of three species of sandflies

Soil analysis was carried out on samples obtained from a breeding-site of the sandflies Phlebotomus perniciosus Newstead, P. perfiliewi perfiliewi Parrot and Sergentomyia minuta (Rondani) in Sardinia. Soil texture, pH, CaCo3, organic matter and water content showed no correlation with the number of sandflies that emerged from the places where soil samples were taken. It appears that pre-imaginal stages of these phlebotomine sandflies are associated with a relatively stable, cool and humid environment protected from sunshine and rain, rich in clay and in organic nitrogen.     

Morphology and ultrastructure of spiracles in phlebotomine sandfly larvae

The morphology and ultrastructure of the larval spiracle system of three phlebotomine sandfly species, Phlebotomus perniciosus, P. perfiliewi and P. papatasi, were examined by scanning (SEM) and transmission (TEM) electron microscopy and by confocal scanning laser microscopy (CSLM). During larval development, thoracic and abdominal spiracles show considerable modifications. In fourth instar larvae, the spiracles consist of a plate with a sclerotized central portion and a peripheral circle of papillae. The latter is distinctive in the larvae of P. papatasi, which are readily distinguished from the other species. Opening clefts across the papillae communicate with an internal chamber that encircles an electrondense plug. Many cylindrical projections cross the chamber, uniting the central plug with the larval body, forming an air filter. Spiracular development in successive larval instars has both a taxonomic and adaptive value.     

Phylogenetic analysis of Phlebotomus species belonging to the subgenus Larroussius (Diptera, psychodidae) by ITS2 rDNA sequences

In the genealogy of Phlebotomus (Diptera: Psychodidae), morphological analyses have indicated that the subgenus Larroussius is a monophyletic group which is most closely related to the subgenera Transphlebotomus and Adlerius. We conducted a phylogenetic analysis of the relationships among six representative species of the subgenus Larroussius and one species representatitive of the Phlebotomus subgenus, assessing sequences of the Second Internal Transcribed Spacer (ITS2) of the ribosomal RNA (rRNA). Three of the species (P. perniciosus, P. ariasi and P. perfiliewi perfiliewi) were collected in different parts of the Mediterranean area. The trees estimated from parsimony and neighbour-joining analyses supported the monophyly of the Larroussius subgenus inferred from the morphological analysis. According to our data, P. ariasi may be a sister group to the rest of the Larroussius subgenus, although additional sequence data are needed to confirm this observation. Our results suggest that P. perniciosus and P. longicuspis are distinct species, in spite of the fact that there are only slight morphological differences. The strict congruence between the phylogeny of the Larroussius subgenus inferred from the ITS2 sequences and that based on morphological studies further confirmed the ability of the spacer sequence to identify recently-derived affiliations.