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PubMed2006

Laboratory testing of the insect repellents IR3535 and DEET against Phlebotomus mascittii and P. duboscqi (Diptera: Psychodidae).

The efficacy of the repellent IR3535 was tested and compared with DEET using two laboratory colonies of phlebotomine sand flies, Phlebotomus mascittii and Phlebotomus duboscqi. P. duboscqi, originating from Senegal, is a proven vector of Leishmania major. P. mascittii (Corsican origin) has never been demonstrated to be a vector of leishmaniasis, but it was reported in 2004 as an anthropophilic species in Germany associated with autochthonous visceral leishmaniasis in people and animals. IR3535 and DEET were tested at an effective concentration of 10% in ethanol on two (one male, one female) human volunteers. The mean protection time against P. duboscqi was 5.9h for both repellents, against P. mascittii 10.4 and 8.8 h for IR3535 and DEET, respectively.

Animals↗
PubMed2006

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

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

Animals↗
PubMed2000

Identification of cutaneous Leishmaniasis vectors, Phlebotomus papatasi and P. duboscqi using random amplified polymorphic DNA.

Phlebotomus papatasi and P. duboscqi are two closely related, morphologically similar sandfly species that are established vectors of zoonotic cutaneous leishmaniasis in the Old World. We have developed a Random Amplified Polymorphic DNA (RAPD) method to find species-specific DNA profiles of these two species. It was found that using a single 10-mer primer a 'species specific' amplification band of about 0.49 kb was produced in all specimens of P. duboscqi while it was absent in P. papatasi. The 0.49 kb diagnostic band was consistently present in both males and females of P. duboscqi. The suitability of this primer for Phlebotomus species identification will help to find the true vector-parasite relationship in the epidemiology of leishmaniasis, particularly in the African countries where both species are prevalent.

Animals↗
PubMed2000

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.

Animals↗
PubMed2002

Characterization of the lectin from females of Phlebotomus duboscqi sand flies.

Lectin from females of the important sand fly vector, Phlebotomus duboscqi (Diptera: Psychodidae), was isolated by immunoaffinity chromatography using a minicolumn with immobilized anti-lectin immunoglobulins. Carbohydrate-binding specificity of active fractions corresponded to that of midgut and salivary gland lysates. Haemagglutination was inhibited by d-glucosamine, d-galactosamine and d-mannosamine. The homogeneity and molecular mass of the purified lectin was examined by SDS/PAGE in both reducing and nonreducing conditions. The active fractions showed one band strongly stained by Coomassie blue or silver nitrate; the molecular mass of the lectin was 42 kDa under nonreducing and 44 kDa under reducing conditions. SDS/PAGE of active fractions from the gel filtration revealed four to six protein bands, but the 42/44-kDa protein present in all active fractions was the only component reacting with specific antibodies in Western blots. Localization of the lectin in the gut of females was studied using indirect immunofluorescence on sections. The positive reaction of specific antibodies was localized in the lumen and along the microvillar surfaces of epithelial cells. The lectin was partially sequenced and characterized by MS. Peptide maps were obtained by MALDI-TOF MS, and several sequence tags were identified from tandem mass spectra on an ion trap. These sequences displayed high similarity to salivary protein precursors previously identified in a cDNA library of the sand flies Phlebotomus papatasi and Lutzomyia longipalpis. Two main hypotheses on the role of female lectin in Leishmania development are discussed.

Animals↗
PubMed2004

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.

Animals↗
PubMed2003

Host-specific Wolbachia strains in widespread populations of Phlebotomus perniciosus and P. papatasi (Diptera: Psychodidae), and prospects for driving genes into these vectors of Leishmania.

A single strain of Wolbachia (alpha-proteobacteria, Rickettsiales) was found in widespread geographical populations of each of two Phlebotomus species, within which there was no indication of 'infectious speciation'. The two strains were identified by sequencing a fragment of wsp (a major surface protein gene), amplified by polymerase chain reaction from DNA extracted from the body parts of individual sandflies. Infection rates were high in the males and females of both sandflies, but they were lower for the B-group wPrn strain of Wolbachia in Phlebotomus perniciosus Newstead (60.3% overall) than for the A-group wPap strain in P. papatasi (Scopoli) (81.7%). Infections were frequent in the thorax, where Leishmania develops infective forms, as well as in the abdomen, where Wolbachia must infect the reproductive tissues to ensure its vertical transmission. These findings were related to knowledge of the population biology of Wolbachia in other insects, leading to the conclusion that this endosymbiont could be useful for driving transgenes through wild populations of both sandflies. This will require characterizing the cytoplasmic incompatibility phenotypes of Wolbachia-sandfly combinations, as well as estimating for them the incidence of paternal transmission and the fidelity of maternal transmission. Paternal transmission is one explanation for finding a single Wolbachia strain associated with all mitochondrial haplotypes and lineages of each sandfly species. However, this distribution pattern could also result from multiple horizontal transmissions or the failure of wsp to provide strain markers.

Animals↗
PubMed1988

The peritrophic membrane of the female sandfly Phlebotomus papatasi.

The ultrastructure of the peritrophic membrane of the female sandfly Phlebotomus papatasi has been studied at various times after blood meals. The membrane begins to form within four hours of the blood meal with the secretion by the entire midgut epithelium of an electron-dense amorphous material. Subsequently, the membrane is stabilized and strengthened by the production of a layer of irregular chitinous microfibres, the whole membrane then forming a complete and resilient sac apparently unaffected by boiling 9 M potassium hydroxide. The membrane appears redundant 48 hours after the blood meal and fragments, possibly as a result of chitinase activity. The membrane's main functions are probably the prevention of clogging of the microvillous brush border by the blood meal and the confinement of large proteins, particularly serum trypsin inhibitors, to the endoperitrophic space while allowing sandfly proteases access to the blood meal periphery. Blood is not required to stimulate membrane production. Saline taken by blood feeding into the midgut also stimulates membrane formation. Phlebotomus papatasi females may lack an efficient anticoagulant, at least in the midgut, as blood meals frequently include fibrin clots.

Animals↗
PubMed1993

Studies on the Phlebotomus fauna of Mount Elgon, Kenya.

In a study of sandflies in 229 of 237 caves found on the slopes of Mount Elgon, Kenya, the Phlebotomus (Larroussius) species P. pedifer and P. elgonensis were found distributed at different altitudes. The females of these two species, which were the only ones of the genus Phlebotomus encountered in the study, were morphologically indistinguishable using existing techniques at the time. On the basis of the morphological identification of the males, however, P. pedifer occurred mainly at the lower altitudes (1750-1900 m) while P. elgonensis predominated at higher altitudes (2300-2600 m). The man-biting activities and the presence of promastigotes in the guts of female sandflies were restricted to lower altitudes (below 1900 m), i.e. the area of maximum distribution of P. pedifer and of cases of cutaneous leishmaniasis. This supports the suggestion that P. pedifer is a vector of cutaneous leishmaniasis on Mount Elgon.

Altitude↗
PubMed1994

A study of phlebotomine sandflies (Diptera:Psychodidae) in Yunnan Province. III. Phlebotomus (Adlerius) fengi sp. nov.

Among the phlebotomine specimens collected from Yunnan, China, in 1984, a new species of the subgenus Adlerius Nitzulescu 1931 was identified. The name Phlebotomus fengi is proposed for this species, in memory of the late medical entomologist, Professor Lan Chow Feng. Although generally similar to P. longiductus, P. fengi had distinct pharyngeal armature, style spines, paramere and aedeagus. Phlebotomus fengi may be a vector of visceral leishmaniasis in southern China.

Animals↗
PubMed1996

A new sandfly species, Phlebotomus (Larroussius) ashfordi (Diptera, Psychodidae) from Ethiopia, previously confused with P. (L.). aculeatus.

The sandfly subgenus Phlebotomus (Larroussius) contains species which transmit parasites causing cutaneous and visceral leishmaniasis in East Africa, including Ethiopia. The Gewasha caves in central Ethiopia harbour a new species (Phlebotomus ashfordi) belonging to the subgenus Larroussius and previously identified as P. (L.) aculeatus Lewis, Minter and Ashford. Although generally similar to P. (L.) aculeatus and P. (L.) elgonensis Ngoka, Madel and Mutinga, the species is distinct in eight male characters and in the female spermatheca. Given its taxonomic affinity to known vectors of cutaneous leishmaniasis and the presence of disease in the Gewasha caves area, the medical importance of this species needs to be assessed.

Animals↗
PubMed1998

Environmental determinants of the distribution of Phlebotomus orientalis in Sudan.

Despite its importance as a vector of visceral leishmaniasis in Sudan, the ecology of Phlebotomus orientalis is still poorly understood. The results of a ground-based survey and a geographical-information-system (GIS) study, carried out to investigate the environmental determinants of the distribution of P. orientalis in the wooded areas of the central savannah belt of Sudan, are described here. The survey, carried out in April-June 1996, consisted of a collection of sandflies over two consecutive nights at each of 44 study sites, using three CDC, miniature, light traps at each site. During the survey, the ecology of each site was described. Phlebotomus orientalis was caught at 17 of the sites. Environmental data on the collection sites (rainfall, minimum and maximum temperatures, soil class, vegetation and land-surface-temperature indices) were extracted from a range of sources of digital data collected by satellites in the National Ocean and Atmospheric Administration's series. These data were then analysed, to ascertain which variables were significantly associated with sites positive for P. orientalis. In line with the results of previous studies, P. orientalis was found to have a significant association with the presence of the tree species Acacia seyal and Balanites aegyptiaca and with the black cotton (vertisolic) soils of eastern Sudan. The positive sites were found to have significantly higher annual mean maximum and minimum daily temperatures than the negative sites and the annual mean maximum normalized-difference vegetation index (NDVI) value was also found to be significantly higher in these sites than in sites where no P. orientalis were found.

Animals↗
PubMed1992

Leishmania infantum MON-98 isolated from naturally infected Phlebotomus langeroni (Diptera: Psychodidae) in El Agamy, Egypt.

Leishmania infantum zymodeme MON-98 was isolated in El Agamy, Egypt. A total of 15 (1.07%) Leishmania-like infections in the anterior midgut and in the head was found in 1,405 Phlebotomus langeroni (Nitzulescu); none of 1,785 Phlebotomus papatasi (Scopoli) was found infected. Four of the 15 cultures (26.7%) were indistinguishable from a reference L. infantum MON-98 strain using cellulose acetate electrophoresis. The isolation and identification of L. infantum MON-98 from naturally infected P. langeroni confirms that this species of sand fly is the vector of visceral leishmaniasis in El Agamy.

Animals↗
PubMed2004

Altitudinal variation in morphometric and molecular characteristics of Phlebotomus papatasi populations.

Four populations of the phlebotomine sandfly Phlebotomus (Phlebotomus) papatasi (Scopoli) (Diptera: Psychodidae), in different ecoregions at altitudes between 368 and 1117 m in the Sanliurfa Province of Turkey, were compared using morphometric and isoenzyme analyses. A similarity phenogram obtained from allozyme data showed that heterozygosity was extremely low, particularly for the alleles which were found to be completely fixed in populations at Hamdun (HMD) and Alitas (ALT). Populations at Akcakale (AKL) and ALT branched as a separate group from populations at Hayatiharrani (HHR) and HMD. The ALT population at the highest altitude (1117 m), and the HHR population (488 m) were clustered distinctly when linear measurements of 46 morphological characteristics were examined. A UPGMA (unweighted pair-group method using arithmetic averages) phenogram also showed that ALT and HHR clustered separately, whereas AKL and HMD formed another group.

Altitude↗
PubMed1992

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.

Animals↗
PubMed2006

High degree of conservancy among secreted salivary gland proteins from two geographically distant Phlebotomus duboscqi sandflies populations (Mali and Kenya).

BACKGROUND: Salivary proteins from sandflies are potential targets for exploitation as vaccines to control Leishmania infection; in this work we tested the hypothesis that salivary proteins from geographically distant Phlebotomus duboscqi sandfly populations are highly divergent due to the pressure exerted by the host immune response. Salivary gland cDNA libraries were prepared from wild-caught P. duboscqi from Mali and recently colonised flies of the same species from Kenya. RESULTS: Transcriptome and proteome analysis resulted in the identification of the most abundant salivary gland-secreted proteins. Orthologues of these salivary proteins were identified by phylogenetic tree analysis. Moreover, comparative analysis between the orthologues of these two different populations resulted in a high level of protein identity, including the predicted MHC class II T-cell epitopes from all these salivary proteins. CONCLUSION: These data refute the hypothesis that salivary proteins from geographically distinct populations of the same Phlebotomus sandfly species are highly divergent. They also suggest the potential for using the same species-specific components in a potential vector saliva-based vaccine.

Africa, Eastern↗
PubMedDate not supplied

Numerical taxonomy of Old World Phlebotominae (Diptera: Psychodidae). 1. Considerations of morphological characters in the genus Phlebotomus Rondani & Berté 1840.

Numerical analyses (correspondence analysis, ascending hierarchical classification, cladistic approach) were applied to the morphological characters of the adults of the genus Phlebotomus Rondani & Berté 1840. They confirm the reliability of the classic classifications, and also redefine the taxonomic and phylogenetic position of certain taxa. Thus, Spelaeophlebotomus Theodor 1948. Idiophlebotomus Quate & Fairchild 1961 and Australophlebotomus Theodor 1948 deserve generic rank. Among the vectors of leishmaniasis, the subgenus Phlebotomus Rondani & Berté 1840 is probably ancient. The results attribute an intermediate taxonomic and phylogenetic position to the taxa Euphlebotomus Theodor 1948 and Anaphlebotomus Theodor 1948, and reveal the probable artificial nature of the latter. The comparatively large numbers of species of subgenera Paraphlebotomus Theodor 1948, Synphlebotomus Theodor 1948 and, above all, Larroussius Nitzulescu 1931 and Adlerius Nitzulescu 1931, suggest that they are relatively recent. The development of adult morphological characters, the validity of their use in taxonomy and proposals for further studies are discussed.

Animals↗
PubMed2003

Phlebotomus (Paraphlebotomus) gemetchi (Diptera: Psychodidae), a new sand fly species from Ethiopia.

The subgenus Paraphlebotomus of Phlebotomus includes several sand fly vectors of cutaneous and visceral leishmaniasis. A new species of the subgenus Paraphlebotomus, named Phlebotomus gemetchi is described from the volcanic blisters by the shore of lake Beseka near Metahara, Upper Awash Valley, in the Rift Valley of eastern Ethiopia. The presence of one distinct character in the male genitalia (a bifurcating 'y'-shaped spine at the middle of the style) and three characters in the female (lengths of the fourth antennal and palpal segments and labrum) easily distinguish it from the other Afrotropical species recorded in East Africa including Ethiopia. The new species is 1 of 5 species representing the subgenus in Ethiopia as well as the rest of the Afrotropical Region. The distribution and medical importance of this sand fly species requires further investigation.

Animals↗