About the presence of Phlebotomus sergenti Parrot, 1917 (Diptera: Psychodidae) in Eastern Sicily, Italy

The note reports the data of a three-year sand fly investigation (1997-99) carried out in Eastern Sicily (Italy) with the aim to study the distribution of Phlebotomus sergenti. The survey involved a densely inhabited area at the foot of Mount Etna and the area of Iblei mounts. A total of 9,095 sand flies, of which 63.4% males, were captured. Five species belonging to the genus Phlebotomus (P. perniciosus, P. perfiliewi, P. neglectus, P. sergenti and P. papatasi) and one to the genus Sergentomyia (S. minuta) were identified. Both the prevalence and distribution of the species were different within the two areas studied. In Mount Etna area, P. perniciosus (77.7%) was the prevalent species followed by S. minuta (19.8%), P. sergenti (2.0%), P. neglectus (0.3%) and P. papatasi (0.2%). While in Iblei mounts region S. minuta (84.5%) showed the highest prevalence, followed by P. perniciosus (14.4%), P. perfiliewi (0.9%) and P. neglectus (0.1%). Here, P. sergenti was a very rare species (< 0.02). P. sergenti was mostly associated to domestic habitats of peri-urban and urban zones located between two and 750 m a.s.l. The density values of P. sergenti, expressed as number of specimens/m2 of sticky trap, were between 0.3 and 5.5 with the highest value in the hilly collecting sites. The low observed abundance of P. sergenti does not allow to draw any prediction on the role that the species could play in the transmission of leishmaniasis in Sicily.     

The development of Psychodiella sergenti (Apicomplexa: Eugregarinorida) in Phlebotomus sergenti (Diptera: Psychodidae)

Psychodiella sergenti is a recently described specific pathogen of the sand fly Phlebotomus sergenti, the main vector of Leishmania tropica. The aim of this study was to examine the life cycle of Ps. sergenti in various developmental stages of the sand fly host. The microscopical methods used include scanning electron microscopy, transmission electron microscopy and light microscopy of native preparations and histological sections stained with periodic acid-Schiff reaction. Psychodiella sergenti oocysts were observed on the chorion of sand fly eggs. In 1st instar larvae, sporozoites were located in the ectoperitrophic space of the intestine. No intracellular stages were found. In 4th instar larvae, Ps. sergenti was mostly located in the ectoperitrophic space of the intestine of the larvae before defecation and in the intestinal lumen of the larvae after defecation. In adults, the parasite was recorded in the body cavity, where the sexual development was triggered by a bloodmeal intake. Psychodiella sergenti has several unique features. It develops sexually exclusively in sand fly females that took a bloodmeal, and its sporozoites bear a distinctive conoid (about 700 nm long), which is more than 4 times longer than conoids of the mosquito gregarines.     

The phlebotomine sandflies of Portugal. XIII--Occurrence of Phlebotomus sergenti Parrot, 1917 in the Arrabida leishmaniasis focus

In a survey carried out during the summer in 2002 and 2003, in the canine and vulpine leishmaniasis focus of Arrabida, 665 phlebotomine sandflies were caught. 13.83% were P. ariasi, 58.65% P. pemiciosus, 0.45% P. sergenti and 27.07% S. minuta. Despite the low abundance, the finding of the three adults P. sergenti (two males in Aldeia Grande and one female in Quinta da Ramada) confirm the colonization of the Arrabida leishmaniasis focus by this species, which presence had been previously reported but thought accidental. The abundance of P. ariasi observed at this time is significantly different from that in previous years (Pires, 1984 and Fernandes pers. com., 1994). The occurrence of P. sergenti in this region, in association with the decrease in abundance of P. ariasi, may reflect an increased aridness of this region, a consequence of current climate and environmental changes.     

Phlebotomus from Madagascar (Diptera: Psychodidae). III--Description of Phlebotomus (Anaphlebotomus) fontenillei n. sp

The male of Phlebotomus (Anaphlebotomus) fontenillei n. sp. is described from Namoroka area (Madagascar). Its belongs to the subgenus Anaphlebotomus: style with four spines, coxite without basal process and paramere with two branches. It shares with P. berentiensis an original and exclusive antennal formula: 2/III-XII which distinguishes them from P. fertei. P. fontenillei n. sp. differs mainly from P. berentiensis by about 40 setae in tuft on the ventral face of the coxite, the length of the genital ducts and the position of the spines on the style. Sequence of the second internal transcribed spacer (ITS2) of the ribosomal DNA (rDNA) is very informative: the male of P. fontenillei n. sp. cannot be linked to the female of P. huberti (male unknown) regarding the size of amplified DNA fragment (459 bp versus 600 respectively) and the high degree of variability. There are few differences (10 mutations) between the sequences of P. fontenillei n. sp. and P. berentiensis which are closely related species.     

Laboratory estimation of degree-day developmental requirements of Phlebotomus papatasi (Diptera: Psychodidae).

Cutaneous leishmaniasis is one of the most important vector-borne endemic diseases in Turkey. The main objective of this study was to evaluate the influence of temperature on the developmental rates of one important vector of leishmaniasis, Phlebotomus papatasi (Scopoli, 1786) (Diptera: Psychodidae). Eggs from laboratory-reared colonies of Phlebotomus papatasi were exposed to six constant temperature regimes from 15 to 32 degrees C with a daylength of 14 h and relative humidity of 65-75%. No adult emergence was observed at 15 degrees C. Complete egg to adult development ranged from 27.89 +/- 1.88 days at 32 degrees C to 246.43 +/- 13.83 days at 18 degrees C. The developmental zero values were estimated to vary from 11.6 degrees C to 20.25 degrees C depending on life stages, and egg to adult development required 440.55 DD above 20.25 degrees C.     

ITS 2 sequences heterogeneity in Phlebotomus sergenti and Phlebotomus similis (Diptera,Psychodidae): possible consequences in their ability to transmit Leishmania tropica

An intraspecific study on Phlebotomus sergenti, the main and only proven vector of Leishmania tropica among the members of the subgenus Paraphlebotomus was performed. The internal transcribed spacer 2 (ITS2) sequences of 12 populations from 10 countries (Cyprus, Egypt, Italy, Lebanon, Morocco, Pakistan, Portugal, Spain, Syria, and Turkey) were compared. Samples also included three species closely related to P. sergenti: Phlebotomus similis (three populations from Greece and Malta), Phlebotomus jacusieli and Phlebotomus kazeruni. Our results confirm the validity of the taxa morphologically characterised, and imply the revision of their distribution areas, which are explained through biogeographical events. At the Miocene time, a migration route, north of the Paratethys sea would have been followed by P. similis to colonise the north of the Caucasus, Crimea, Balkans including Greece and its islands, and western Turkey. Phlebotomus sergenti would have followed an Asiatic dispersion as well as a western migration route south of the Tethys sea to colonise North Africa and western Europe. This hypothesis seems to be well supported by high degree of variation observed in the present study, which is not related to colonisation or to intra-populational variation. Two groups can be individualised, one oriental and one western in connection with ecology, host preferences and distribution of L. tropica. We hypothesise that they could be correlated with differences in vectorial capacities.     

Free amino acids in wild-caught Phlebotomus perniciosus and P. perfiliewi (Diptera: Psychodidae).

Free amino acids occurring in wild Phlebotomus perniciosus and P. perfiliewi were analyzed using high performance liquid chromatography. For both species the total concentration of amino acids, per unit weight, is over three times greater in females than in males. Qualitative inter-sex differences in amino acid compositions were also discovered: the most significant involving higher proportions of taurine and gamma-aminobutyric acid in females and higher proportions of arginine and glutamic acid in males. Although amino acid composition is consistent between females of both species, there are significant differences between males. The most important difference between males is an increase in tyrosine levels, at the expense of taurine, in P. perniciosus (as compared with P. perfiliewi). It is suggested that the intra- and inter-specific variation in amino acid composition identified in these studies could be due to ecological factors, such as differences in the sandflies natural amino acid sources (honeydew and/or plants), although a physiological explanation is not ruled out.     

Ultrastructure of the seminal vesicle of Phlebotomus perniciosus Newstead (Diptera, Psychodidae)

The seminal vesicles of Phlebotomus perniciosus were investigated by light microscopy, confocal scanning laser microscopy and by scanning and transmission electron microscopy. They have a complex structure, and three different morphological compartments called A, B and C are distinguished on the basis of their position and fine structure. Compartment A is continuous with the vasa deferentia and consists of a cylindrical wall limiting a lumen in which the spermatozoa are stored. Compartment B is hemispherical and surrounds compartment A like a muff. Compartment C constitutes an external coat surrounding A and B. The epithelial cells of each compartment are characterized by morphologically different secretory granules. The ultrastructural features of these cells are described and their role in sandfly reproductive biology is discussed.     

Malformations of the genitalia in male Phlebotomus papatasi (Scopoli) (Diptera: Psychodidae)

Phlebotomus papatasi ( Scopoli, 1786 ) (Diptera: Psychodidae) is a major vector of Leishmania major (Kinetoplastida: Trypanosomatidae), a causative agent of zoonotic cutaneous leishmaniasis. Morphological characters of sand fly genitalia are key indicators for species identification. Various anomalies affecting male genitalia have been previously described. We take advantage of a large sand flies survey conducted in 32 stations in Central and Southern Morocco to systematically quantify the prevalence and spatial distribution of malformations affecting the genitalia of P. papatasi. Among 597 examined males, 122 were abnormal (20.4%). Malformations were widespread and largely concerned the number of spines in the lateral lobes and in the styles. Asymmetrical anomalies in lateral lobes were common. Correspondence analysis of our results highlighted the symmetrical anomalies observed in the lateral lobes, and abnormal styles of the male genitalia were found to be associated with environmental disturbances since they were prevalent in sewage dumps.     

Phlebotomus longipes Parrot & Martin (Diptera: Phlebotomidae) and a new related species

Hitherto it was considered that Phlebotomus longipes Parrot & Martin occurred in Ethiopia, Kenya and the Sudan and was the vector of Leish-mania tropica (Wright) in Ethiopia and in one area of Kenya. Study of additional specimens indicates that those from Kenya, the Sudan and one area of Ethiopia represent a new species. Both species show local variation.     

First description of the male of Phlebotomus betisi Lewis and Wharton, 1963 (Diptera: Psychodidae)

The male of Phlebotomus (Larroussius) betisi is described from Malayan caves. Several males have been caught in association with P. betisi females. Males and females have been associated by ecology, biogeography, morphology and molecular biology (homology of the ND4 mtDNA sequences).     

Comparative demography of the sand fly Phlebotomus papatasi (Diptera: Psychodidae) at constant temperatures.

We measured reproductive and population parameters of adult sand flies, Phlebotomus papatasi (Scopoli, 1786) (Diptera: Psychodidae), in environmental chambers maintained at temperatures of 15, 18, 20, 25, 28, and 32 degrees C. Based on cohorts of adults at each temperature regime, horizontal life tables were constructed using established laboratory colonies initiated from specimens collected in Sanliurfa Province, southeastern Anatolia, Turkey. The fecundity and longevity of the insects were both highly variable, depending on the temperature. At 15 degrees C, all of the cohort females died before laying eggs, so the construction of a life table for this temperature regime was not possible. Within a range of 18 to 32 degrees C, the longevity of adult P. papatasi increased as the temperature decreased; at 15 degrees C, the mean survival times of females and males were 19.04 +/- 6.94 days (9-35) and 17.84 +/- 7.11 days (9-33), respectively. While the highest number of eggs was found in the cohort at 28 degrees C (44.08 +/- 7.79), this was only 3.60 +/- 1.55 in the cohort at 32 degrees C and 2.8 +/- 0.9 in the cohort at 18 degrees C. This result showed that extreme temperatures negatively affect the fecundity of this species. The cohort reared at 28 degrees C exhibited the highest intrinsic rates of population increase (r(m)) for P. papatasi. The r(m) ranged from 0.098 at 28 degrees C to 0.007 at 18 degrees C. The cohort placed at 28 degrees C was found to be significantly different (P < 0.01) from the other cohorts producing the fewest progeny in terms of net reproductive rate, R(0), (15.87). The values for mean generation time (T) were estimated to vary from 36 days to 271 days depending on temperature. Principal Component Analysis (PCA) confirmed results from the previous studies that the cohort at 28 degrees C orientated and clustered as a distinct group along the first two PCs.     

Isolation and characterization of microsatellite loci in the sand fly Phlebotomus papatasi (Diptera: Psychodidae)

Phlebotomus papatasi is a proven vector of Leishmania major which is one of the causative agents of cutaneous leishmaniasis in the Old World. Although it has a wide geographical range, its population structure is not yet well understood. In an effort to better understand the population dynamics of this vector, we developed a panel of di‐ and trinucleotide microsatellite markers, using a magnetic bead hybridization enrichment protocol. These microsatellite loci showed three to seven alleles with an expected heterozygosity range between 0.702 and 0.876. The level of polymorphisms found in this study suggests that these microsatellite loci can be used for population analysis of P. papatasi.     

Ultrastructure of reproductive accessory glands in the female sandfly Phlebotomus perniciosus Newstead (Diptera : Psychodidae)

The paired female accessory glands of Phlebotomus perniciosus (Diptera : Psychodidae) were investigated by light microscopy, and by scanning and transmission electron microscopy. These glands undergo morphological and functional changes during oocyte development. After the blood meal, the monostratified glandular epithelium differentiates and starts to secrete. Well-developed rough endoplasmic reticulum, Golgi complexes, and membrane-bounded exocytic vesicles suggest that these secretory cells are involved in protein synthesis. As the secretory cells differentiate, the glandular lumen increases in size and fills with secretory material, consisting of globular granules of different sizes in an amorphous electron-dense matrix. The granules have an electron-translucent core and an electron-dense cortex. The morphological characteristics of the glandular epithelium and the functional role of the glands are discussed in relation to their possible contribution to the reproductive process.