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.     

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.     

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.     

Dispersal of phlebotomine sand flies (Diptera: Psychodidae) in a Colombian focus of Leishmania (Viannia) braziliensis.

Dispersal of five species of phlebotomine sand flies was studied in a coffee plantation near Arboledas, Colombia, by mark-release-recapture studies using fluorescent powders. The estimated recapture rate for males of Lutzomyia shannoni marked and released during the day was 28.1%, significantly higher than that for all other species (p < 0.05). Recapture rate of female Lu. shannoni was 9.5%, and no females of the other four species were recovered. This suggests either that Lu. shannoni is a more sedentary species than the others, or that the large trees on which these insects were captured and recaptured function as foci of lekking behaviour in males. The high recapture rates of females of this species may indicate that oviposition occurs in close proximity to the bases of these trees. Although most marked sand flies were recaptured within 200 m of their release point, a single female Lu. gomezi was recovered 960 m away 36 h after release. This suggests that the dispersal capacity of Lutzomyia species may be greater than has been thought, an important consideration in future control programs directed against these insects in Leishmania-endemic areas.     

Ecology of phlebotomine sand flies (Diptera: Psychodidae) in a focus of Leishmania (Viannia) braziliensis in northeastern Colombia.

The phlebotomine sand fly fauna of two coffee plantations in a Leishmania-endemic area of Norte de Santander, Colombia was studied. Regular insect collections using a variety of methods were made for three and a half years. Information was obtained on diurnal resting sites, host range and seasonal abundance for 17 species, of which five (Lutzomyia spinicrassa, Lu. serrana, Lu. shannoni, Lu. ovallesi and Lu. gomezi) were far more numerous than the others, anthropophilic and present throughout the year. The behaviour of these and the remaining 12 species is discussed in relation to their potential role in transmission of Leishmania (Viannia) braziliensis in the area.     

Phlebotomine sand flies (Diptera: Psychodidae) associated with changing patterns in the transmission of the human cutaneous leishmaniasis in French Guiana

Between March 2000 and December 2001 a survey of the sand flies (Diptera: Phlebotominae) of French Guiana was carried out during 14 nights of captures with CDC light-traps and Malaise traps, and resulted in the collection of 2245 individuals of 38 species. The most abundant species were Lutzomyia (Trichophoromyia) ininii Floch & Abonnenc, Lu.(Psychodopygus) squamiventris maripaensis Floch & Abonnenc, and Lu .(Nyssomyia) flaviscutellata Mangabeira. Half of the collected sand flies females were dissected under field conditions and five species were found harboring Leishmania-like parasites. The Leishmania (Kinetoplastidae: Trypanosomatidae) species were identified by molecular typing, and for the first time Lu. (Nys.) flaviscutellata was found harboring Leishmania (Viannia) guyanensis and Lu. (Tri) ininii harboring unknown Leishmania. The first record for French Guiana of Lu. (Psy.) squamiventris maripaensis harboring L. (V.) naiffi, was also reported. The patterns of diversification of the human cutaneous leishmaniasis transmission in French Guiana are discussed.     

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.     

Radiation of the Oriental phlebotomine sand flies (Diptera: Psychodidae)

Abstract The historical biogeography of phlebotomine sand fly taxa Hertigia, Warileya, Phlebotomus (Idiophlebotomus), P. (Spelaeophlebotomus), P. (Anaphlebotomus), and P. (Euphlebotomus) and the Phlebotomus (Euphlebotomus) argentipes species complex was investigated using phylogenetic inference from comparative genital morphology, distribution of ancestral taxa, fossil evidence and geological age. Idiophlebotomus and Euphlebotomus occur in the Oriental region with one species from northeast Australia, whereas Anaphlebotomus occurs both in the Afro‐tropical and Oriental regions. These disjunct distribution patterns across the Oriental region and the present day distribution are likely to be vicariance due to break of Gondwanaland. Fossil records, extant taxa distribution, phylogenetic analysis of the Old World Phlebotominae and paleogeography suggest that ancestors of Idiophlebotomus and Euphlebotomus originated apparently in the Cimmerian continent of northern margin of Gondwanaland in the early Permian (290 million years ago, MYA) and subsequently radiated in the Mesozoic by tectonic vicariance. The Phlebotomus argentipes species complex occurs in the South and South‐east Asian countries and transmits the protozoan parasite Leishmania donovani that causes visceral leishmaniasis (Kala‐azar) in India, Bangladesh and Nepal. The phylogeography of P. argentipes was caused through vicariance followed by dispersal events from 5O MYA (the Eocene) until the Recent era.     

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.     

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.     

DNA barcoding for identification of sand flies (Diptera: Psychodidae) in India

About 50 species of sand flies have been reported to be prevalent in India. We explored the utility of the DNA barcode approach towards species identification of these medically important insects. A total of 62 specimens belonging to seven morphologically identified species of two genera, Phlebotomus and Sergentomyia, collected from Puducherry Union Territory, Maharashtra and Rajasthan states of India were subjected to the analysis. Neighbor‐joining (NJ) analysis of DNA barcode sequences identified the individuals of seven morphological species into eight distinct species, as presented in the designed NJ tree. This methodology delineated morphologically identified species, S. bailyi, into two genetically isolated groups. Also, this study characterizes DNA barcodes of P. argentipes and P. papatasi, the vector species of leishmaniasis in India, for the first time.     

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.     

Impact of anti-sandfly saliva antibodies on biological aspects of Phlebotomus papatasi (Diptera: Psychodidae), vector of cutaneous leishmaniasis

Sandflies are the main vectors of Leishmania parasites in tropical and subtropical areas. The immunization of vertebrate hosts with vector components through repeated bites may offer an alternative method for sandfly control. Aliquots of female Phlebotomus papatasi (Scopoli) (Diptera: Psychodidae) were weekly blood fed on 12 individual hamsters throughout 18 successive weeks. Significant biological and biochemical changes resulting from antibodies developed by immunized host sera against repeated biting were observed in sandfly females. Blood feeding and fertility rates of females significantly gradually declined to the end of the study period. No appreciable difference was observed in mortality rates among flies repeatedly fed on individual hamsters throughout weeks 9 and 18, compared to flies fed on naïve hamsters. Total salivary gland proteins of female sandflies were compared to proteins in sera of sensitized hamsters. SDS-page revealed bands common to both flies and hosts, indicating the development of anti-saliva antibodies in hamster sera. The importance of anti-sandfly saliva antibodies as a potential tool for vector control leading to the interruption of leishmaniasis is discussed.     

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.     

Modification of Disney trap for capture of sand flies (Diptera: Psychodidae: Phlebotominae)

This paper describes the modifications made to the original model of the Disney trap, with a view to easier handling of the same, greater practicability in the collection of sand flies, protection of the animal bait and durability of the trap in the field.     

New moth flies (Diptera: Psychodidae: Psychodinae) from Turkey

Specimens of moth flies (Diptera: Psychodidae: Psychodinae) were sampled in southwest Anatolia and the European part of Turkey between 2003 and 2009. Samples contained nine species new to science: Ulomyia asiaminorica sp. n., Pneumia pellucida sp. n., Berdeniella turcica sp. n., B. aktashi sp. n., Clytocerus thracicus sp. n., Jungiella afyonica sp. n., J. geniculatoides sp. n., J. barlasi sp. n., and Telmatoscopus remotus sp.n. The new species are described and illustrated. Distributional data for a total of 45 species are provided. 63 species of Psychodinae are now known to occur in Turkey.     

Trapping sand flies (Diptera: Psychodidae) in the Emilia-Romagna region of northern Italy.

The efficiency and practicality of two trapping methods for adult Phlebotomine sand flies in two areas of the Emilia-Romagna region (Italy) were evaluated. Suction traps (CO2) and sticky traps (ST) were used to collect sand flies every two weeks, from June to September, 1999, from 16:00 to 07:00. Two CO2 traps were activated at the same time for each area (one with light and one without light), whereas 38 (four with light and 34 without lights) and 48 (four with light and 44 without) sticky traps were activated in Borghi and Longiano, respectively. An Index of Apparent Abundance (IAA) was calculated for each trap type and area. A total of 2,253 sand flies was trapped over the four-month period, with 1,765 collected from Borghi and 488 from Longiano. Phlebotomus perfiliewi was the most abundant species collected, comprising 99.6% and 84.6% of the total flies trapped in Borghi and Longiano, respectively. Other species were also collected within the two areas (Phlebotomus perniciosus and Phlebotomus mascittii) but were not considered for further analyses due to low catches. Significantly more specimens were caught using CO2 than sticky traps and the addition of a light source also improved the catches, however, a significantly greater number of female specimens were collected by a CO2 trap without a light source. Phlebotomus perfiliewi thus appears to show a photophobic reaction in the case of females when trapped using CO2/light attractants.     

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.     

The molecular detection of different Leishmania species within sand flies from a cutaneous and visceral leishmaniasis sympatric area in Southeastern Brazil

Over the last 20 years, there has been an increase in the number of leishmaniasis cases in Brazil. Belo Horizonte (BH) is one of the most highly populated Brazilian cities that is affected by visceral leishmaniasis (VL). The health services in BH are coordinated by a central nucleus that is subdivided into nine sanitary districts. Historically, the highest level of human VL cases was found in the northeast sanitary district (NSD). The objective of our study was to detect Leishmania infection in the phlebotomine sand flies collected in the NSD by dissection and molecular approaches. Following the occurrence of human VL cases in 2005, entomological captures were performed from July 2006-June 2007. Out of the 245 sand flies dissected, only three Lutzomyia longipalpis spp contained flagellates. The female sand flies were grouped into 120 pools according to date, collection site and species, with approximately 10 individual sand flies in each pool. Subsquently, the DNA was extracted and Leishmania spp and other parasites were detected and identified by polymerase chain reaction (PCR) and PCR-restriction fragment length polymorfism. Leishmania infantum was present in at least 19% of the Lu. longipalpis collected, in 3.8% of the Nyssomiya whitmani collected, in 33.3% of the Evandromiya termitophila collected and in 14.3% of the Nyssomiya intermedia collected. When the females of the cortelezzii complex were compared with each other, 3.2% of the females were infected with Leishmania braziliensis, whereas 3.2% of the females were infected with trypanosomatids.