Mitochondrial and Wolbachia markers for the sandfly Phlebotomus papatasi: little population differentiation between peridomestic sites and gerbil burrows in Isfahan province, Iran

In Iran, Phlebotomus papatasi (Scopoli) (Diptera: Psychodidae) is the main vector of Leishmania major Yakimoff & Schokhor (Kinetoplastida: Trypanosomatidae), the causative agent of rural zoonotic cutaneous leishmaniasis. This sandfly is abundant both in villages and in the burrows of the main reservoir host, the gerbil Rhombomys opimus (Licht.) (Rodentia: Gerbillidae). Populations of P. papatasi were sampled from the edges of villages in Isfahan province, using CDC miniature light traps in peridomestic sites and sticky papers placed at the entrances to gerbil burrows. Single peridomestic sites in two northern provinces were also sampled. Individual sandflies were characterized by PCR amplification and sequencing of fragments of their mitochondrial cytochrome b gene and of the wsp gene of endosymbiotic Wolbachia pipientis Hertig (alpha-Proteobacteria: Rickettsiaceae). The distributions of the haplotypes of these two maternally inherited genes were analysed to assess the population differentiation of P. papatasi, knowledge of which will be needed for planning control measures. For the first time these markers were used to characterize P. papatasi from gerbil burrows, and they indicated the absence not only of sympatric cryptic species but also of any long-term differentiation of lineages in different habitats. A single lineage of cytochrome b haplotypes was found, and both sexes in all populations had a high infection rate of the same A-group strain of Wolbachia (wPap). The distributions of cytochrome b haplotypes were consistent with females dispersing more than males, which has been reported for P. papatasi in other countries. The widespread distribution of wPap suggests that Wolbachia could be used to spread transgenes between populations of P. papatasi in different habitats.     

Impact of pyrethroid-impregnated curtains on Phlebotomus papatasi sandflies indoors at Khartoum, Sudan

Laboratory and field investigations were made in an endemic focus of cutaneous leishmaniasis in Khartoum State, Sudan, to evaluate the effects of permethrin-impregnated curtains on the human-biting activity, nocturnal activity and resting behaviour of the vector sandfly Phlebotomus papatasi (Diptera: Psychodidae) indoors. Laboratory bioassays showed that curtains impregnated with 0.5, 1.0 or 1.5 g/m2 permethrin all gave 100% mortality within 24h of exposure of P. papatasi for 3 min. Under natural field conditions, the biting activity indoors and the resting density of P. papatasi were significantly reduced (P<0.001 and P=0.036, respectively) in rooms provided with permethrin-impregnated curtains as compared to control rooms left without curtains or fitted with unimpregnated curtains. No significant difference was found between the numbers of nocturnally active P. papatasi collected in rooms provided with impregnated curtains and rooms left without curtains or provided with unimpregnated curtains (P=0.377). Evidently P. papatasi was not repelled by these doses of permethrin on curtains, but the survival rate of sandflies collected from test rooms provided with permethrin-impregnated curtains was significantly reduced (P=0.036). We conclude that use of permethrin-impregnated curtains may provide a good control method for P. papatasi and other endophilic and/or endophagic sandfly vectors of leishmaniasis.     

Genetic structure of Mediterranean populations of the sandfly Phlebotomus papatasi by mitochondrial cytochrome b haplotype analysis

Phlebotomus papatasi (Scopoli) (Diptera: Psychodidae) is the main vector of Leishmania major Yakimoff & Schokhor; which is the cause of self-limiting cutaneous leishmaniasis in the Old World. This sandfly is found in houses, animal shelters, caves and rodent burrows. It has a large geographical range, which includes the Middle East and the Mediterranean regions. A population analysis of colony and field specimens of P. papatasi was conducted on 25 populations originating from 10 countries. The distribution of haplotypes of the maternally inherited mitochondrial cytochrome b gene were analysed to assess the population differentiation of P. papatasi. Alignment of a 442-basepair region at the 3' end of the gene identified 21 haplotypes and 33 segregating sites from 131 sandflies. The pattern of sequence variations did not support the existence of a species complex. The median-joining network method was used to describe both the origin of the haplotypes and the population structure; haplotypes tended to cluster by geographical location, suggesting some level of genetic differentiation between populations. Our findings indicate the presence of significant population differentiation for populations derived from Syria, Turkey, Palestine, Israel, Jordan and Egypt. Knowledge of population differentiation among P. papatasi populations is important for understanding patterns of dispersal in this species and for planning appropriate control measures.     

DNA hybridizations on squash-blotted sandflies to identify both Phlebotomus papatasi and infecting Leishmania major

Epidemiological field studies on leishmaniasis have been hampered by the laborious, and often inefficient, methods used to assess the rates of infection of sandfly vectors (Diptera; Phlebotominae) by species of the causative disease organisms, protozoal parasites of the genus Leishmania (Kinetoplastida; Trypanosomatidae). We report the rapid and accurate identification of both sandfly vector (Phlebotomus (Phlebotomus) papatasi (Scopoli] and infecting Leishmania major Yakimov & Schokov by DNA hybridizations to squash-blotted sandflies. Large numbers of whole (infected) sandflies can be quickly squashed on to nylon hybridization filters and (following standard procedures) the filter-bound DNA can be hybridized sequentially to cloned, multicopy genomic sequences that are specific for species of Leishmania (kinetoplast DNA) or for the sandfly (ribosomal (r) DNA). Our sandfly probe consists of a 3.2 kb fragment of the intergenic 'non-transcribed' spacer of rDNA of P. papatasi that we have detected only in this species: it is present in all six geographically isolated populations tested (from Tunisia through to India) but cannot be detected in the morphologically similar P. (Phlebotomus) duboscqi Neveu-Lemaire, the vector of Leishmania major south of the Sahara; it also cannot be detected in Phlebotomus species of the subgenera Larroussius and Paraphlebotomus that together with P. papatasi are the dominant man-biting sandflies in north African foci of zoonotic cutaneous leishmaniasis, where (as in many arid regions of western Asia) P. papatasi is believed to be the sole vector of L. major.     

Incrimination of Phiebotomus papatasi as vector of Leishmania major in the southern Jordan Valley

he status of sandflies as vectors of cutaneous leishmaniasis in the southern Jordan Valley was investigated during 1992. Sandflies were collected from domestic habitats and from burrows of Psammomys obesus . Of 686 Phlebotomus papatasi females collected from burrows, fourteen harboured promastigotes in their guts. On the other hand, none of 1446 P.papatasi females collected from domestic habitats were found infected. The highest infection rate (5.5%) was recorded in November at the end of the sandfly season. Six leishmanial stocks isolated from P.papatasi females were typed by cellulose acetate electrophoresis using the six enzymes G6PDH, 6PGDH, PGI, PGM, FK and ME. Five of the leishmanial stocks were identical to a Leishmania major reference strain (MHOMISU/73/5-ASKH). The sixth isolate was a 6PGDH variant of L.major . These findings present the first direct evidence of the role of P.papatasi as a vector of L.major in Jordan.     

Wing size and shape variation of Phlebotomus papatasi (Diptera: Psychodidae) populations from the south and north slopes of the Atlas Mountains in Morocco

The wing shape and size morphology of populations of the medically important phlebotomine sand fly, Phlebotomus papatasi, were examined in two endemic (south of the Atlas Mountains) and nonendemic (north of the Atlas Mountains) foci of cutaneous leishmaniasis by using geometric morphometrics in Morocco. Although it is present in all of Morocco, P. papatasi is the main vector of Leishmania major in only southern part of the Atlas Mountains. There are four major mountain ranges that serve as geographical barriers for species distribution in the study area and at least four gaps were recognized among these barriers. We found statistically significant differences in wing shape morphology between southern and northern populations. Analysis clearly recognized two main groups of populations on both sides of the mountains. The graphical depiction of Principal Component Analysis (PCA) and Canonical Variates Analysis (CVA) confirmed our morphometric study suggesting that the difference in wing morphology between the populations indicates that the population of P. papatasi shows phenotypic plasticity in the study area. According to centroid size analyses, which were used as measures of wing size differences among different sites, the north population of P. papatasi had relatively larger wings than the south population.     

Base-line susceptibility of Phlebotomus papatasi to insecticides

Susceptibility of the leishmaniasis vector sandfly Phlebotomus papatasi (Scopoli) to three insecticides was assessed on a colony originating in the Jordan Valley, Israel. Results are considered to show susceptibility to DDT and permethrin. The results for methoxychlor indicated the possibility of heterogeneity in the population and tolerance towards this compound.     

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.     

Evaluation of juvenile hormone analogues as rodent feed-through insecticides for control of immature phlebotomine sandflies

The juvenile hormone analogues methoprene and pyriproxyfen were evaluated as rodent feed-through insecticides for control of immature stages of the sandfly Phlebotomus papatasi Scopoli (Diptera: Psychodidae). The development and survival of P. papatasi second-instar larvae fed faeces from Syrian hamsters, Mesocricetus auratus, that had been fed a diet containing methoprene (0, 9.788, 97.88 or 978.8 p.p.m.) or pyriproxyfen (0, 9.82, 98.2 or 982 p.p.m.) were evaluated. The faeces of methoprene-treated hamsters greatly reduced the percentage of larvae that pupated at all concentrations tested and prevented adult emergence at all but the lowest concentration (9.788 p.p.m.). Pyriproxyfen prevented both pupation and adult emergence at all concentrations tested. The results of this study suggest that a control strategy using rodent baits containing juvenile hormone analogues to control phlebotomine sandflies that live in rodent burrows and feed on rodent faeces may be possible. As rodent reservoirs and vectors of Leishmania major live in close association in many parts of the Middle East, control of the transmission of the agent of zoonotic cutaneous leishmaniasis may also be possible.     

10. PROCEEDINGS OF THE SEVENTEENTH ANNUAL GENERAL MEETING OF THE S.A. ORNITHOLOGICAL SOCIETY HELD IN THE COMMITTEE ROOM OF THE PUBLIC LIBRARY. JOHANNESBURG,AT 3 P.M., SATURDAY, 8TH MARCH. 1947

Bruderer, B. 1994. Nocturnal bud migration in the Negev (Israel) a tracking radar study. Ostrich 65: 204–212.

The present publication summarizes the methodological possibilities of tracking radar and describes some features of nocturnal migration at two sites in the Negev, which include anwers to basic questions of bird migration. The directions of spring and autumn migration were practically opposite; only the headings in spring indicated some more compensation for stronger westerly winds. The volume of nocturnal spring migration was only about 65% of autumn migration, which may be an indication of mortality outside tie breeding area. Highest densities of migration at the two radar sites in the Negev Highlands (450 m above sea level) and in the Arava Valley (150 m below sea level) indicated flightlevels adjusted to atmospheric conditions aloft, and not to round level. Due to the trade-wind system, the birds heading southward in autumn flew mainly below flew mainly above 1500 m above sea level, while in spring they tended to make use of the anti-trades at higher altitudes. The decisive factor for altitude choice was the speed of tailwind in spring and autumn; other factors, such as temperature, humidity and pressure had no significant influence on the altitude distributions. With respect to the question of non-stop or intermittent flight across large desert areas, the data show that between the eastern deserts of Egypt and the Sinai/Negev complex the nocturnal migrants maintained their schedule of nocturnal flight and diurnal rest. A few exceptions of nocturnal migrants continuing migration at high altitudes into the day were identified mainly as heron- and gull-type birds. The proportion of waders and waterfowl identified by wing-beat pattern in nocturnal migration is nearly the same at both sites, indicating broad-front migration across the desert. The numbers of birds with continuous wingbeats is, however, so large compared to available estimates of waders and waterfowl wintering in Africa that careful reconsideration of the underlying assumptions in the radar and field estimates is necessary.     

Seasonal variation of Leishmania major infection rates in sandflies from rodent burrows in Isfahan province, Iran

Abstract. In preparation for field trials of killed Leishmania major vaccine, natural infections with Leishmania promastigotes were monitored in Phlebotomine sandfly vectors from villages of Borkhar rural district, northeast of Isfahan in central Iran, where Lmajor zymodeme MON-26 (=LON-l) has been identified as causing zoonotic cutaneous leishmaniasis (ZCL). Sandflies were collected and dissected weekly, from burrows of rodent colonies, during the 'sandfly season', June-October 1991. Leptomonad infection rates were 12% of 26 Phlebotomus ansarii , 8% of 280 P.caucasicus , 11 % of 1042 P.papatasi and none of 126 Sergentomyia sintoni , being greatest during late August through September, coinciding with peak activity of the sandflies, 2–3 months before the highest incidence of ZCL human cases in November-December.     

A role for insect galectins in parasite survival

Insect galectins are associated with embryonic development or immunity against pathogens. Here, we show that they can be exploited by parasites for survival in their insect hosts. PpGalec, a tandem repeat galectin expressed in the midgut of the sandfly Phlebotomus papatasi, is used by Leishmania major as a receptor for mediating specific binding to the insect midgut, an event crucial for parasite survival, and accounts for species-specific vector competence for the most widely distributed form of cutaneous leishmaniasis in the Old World. In addition, these studies demonstrate the feasibility of using midgut receptors for parasite ligands as target antigens for transmission-blocking vaccines.     

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.     

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%.     

Control of phlebotomine sandflies in confined spaces using diffusible repellents and insecticides

Abstract.  The control of phlebotomine sandflies (Diptera: Psychodidae), the vectors of leishmaniasis, is directed mostly against adults as larvae develop in unknown or inaccessible habitats. In the current study we tested geraniol, a natural plant-derived product, as a space repellent and the synthetic pyrethroid prallethrin as a diffusible insecticide. Geraniol was dispersed in the air using diffusers with an electric fan and prallethrin was evaporated using electrically heated evaporators. Both substances were tested in inhabited bedrooms and in tents. Geraniol failed to effect significant reductions in the numbers of either Phlebotomus papatasi Scopoli in rooms or Phlebotomus sergenti Parrot in tents. In laboratory experiments, geraniol proved ineffective in preventing sandflies from feeding. By contrast, prallethrin was highly effective in reducing the number of sandflies in rooms as well as in tents. Exposure of sandflies to prallethrin in laboratory experiments caused 97% mortality rates. Both prallethrin and, to a lesser extent, geraniol reduced the number of Culex mosquitoes captured in tents. Electric liquid-vaporizers with 1.5% prallethrin are highly effective in protecting people from sandfly bites in confined spaces and may be useful in combating cutaneous leishmaniasis.     

Sandfly (Diptera, Psychodidae, Phlebotominae) fauna of South-Western Pakistan. 1. Diagnostic morphology of Phlebotomus papatasi (Scopoli), Ph. bergeroti (Parrot) and Ph. salehi (Mesghali)

A survey was conducted to study the morphology of the sandfly fauna in South-Western Pakistan (Balochistan). During the revision of different genera of sandflies the specimens of Phlebotomus papatasi (Scopoli) (N = 720), Ph. bergeroti Parrot (N = 30) and Ph. salehi Mesghali (N = 70) were encountered in various localities. These localities appear to be new records of the subgenus in the literature to date. Ph. bergeroti is reported for the first time from Pakistan and Ph. salehi from Balochistan as well. Characters of these three Pakistanese Phlebotomus are compared with the published data of these species from other countries. Keys for the identification of Pakistanese Phlebotomus are also constructed. Two female Ph. papatasi collected from indoors out of 132 female flies (1.5%) were found positive with flagellate infection in pharynx and midgut. The possible vectorial role of these flies is also discussed. Further surveys are necessary in parts of the country that have not been systematically surveyed.     

Cytoplasmic polyhedrosis viruses in Phlebotomus papatasi inhibit development of Leishmania major

Cytoplasmic polyhedrosis viruses (CPV's) were observed in wild-caught and laboratory-reared Phlebotomus papatasi. Chronic CPV pathology of the midgut, characterized by structural aberrations in the epithelium and the peritrophic membrane, interfered with blood digestion and rendered the sand flies refractory to Leishmania major infections. Rates of natural and artificial L. major infections were inversely correlated to the incidence of CPV infections. The interaction between viruses and protozoan parasites in an insect host is of basic biological interest and in this case may be of significance in the epidemiology of cutaneous leishmaniasis.     

Experimental assessment of the ability of different species of sandfly to transmit the causative agent of visceral leishmaniasis

The authors succeeded in transmission of the visceral leishmaniasis agent (kazakh strain of Leishmania infantum) through the sand flies Phlebotomus longiductus, P. smirnovi and P. papatasi under experimental conditions. Infected P. longiductus and P. smirnovi are capable of preserving the agent and infecting the susceptible mammals after the cessation of the 1st and 2nd gonotrophic cycles. P. papatasi transmitted the agent of visceral leishmaniasis only after the cessation of the 2nd gonotrophic cycle. Under the same conditions the transmission of visceral leishmaniasis agent through P. caucasicus failed. Informative value of characteristics of virtual vectors for differentiation of sand flies as carriers is analysed. A question of the necessity of obtaining additional data, which prove the role of P. caucasicus and P. papatasi as vectors of visceral leishmaniasis agent, is raised.     

Sandfly surveillance and investigation of Leishmania spp. DNA in sandflies in Kosovo

In the past decade, leishmaniasis seems to be re-emerging in Balkan countries. There are serious implications that Kosovo is a visceral leishmaniasis endemic region with autochthonous transmission; nevertheless, surveillance of vectors, reservoirs or the disease is not yet established. Gaining knowledge about sandfly vector species is a prerequisite for the development of a monitoring and control plan in the future. After a long gap in research of over 70 years, sandfly studies in Kosovo were resumed in 2014. During this presence/absence study, nine sandfly species were detected: Phlebotomus papatasi, Ph. perfiliewi, Ph. tobbi, Ph. neglectus, Ph. simici, Ph. balcanicus, Ph. alexandri, Ph. mascittii and Sergentomyia minuta. Three species are new with regard to the fauna of Kosovo – Ph. alexandri, Ph. balcanicus and Ph. mascittii. Besides increased diversity, changes in the number of collected specimens and distribution range of species were recorded, with Ph. neglectus being the most dominant species with the widest distribution. Testing of randomly chosen females for Leishmania spp. DNA resulted the in detection of L. tropica in a specimen of Ph. neglectus. The presence of numerous vector species in the sandfly fauna of Kosovo pose a threat for the re-emergence of vector-borne diseases. Therefore, continuous surveillance is recommended with regular updates on vector distribution and abundance.