Leishmaniasis and phlebotomine sand flies in Oman Sultanate.

There are few data on leishmaniases and sandflies in Oman Sultanate. We carried out an eco-epidemiological study in 1998 in the two main mountains of the country, the Sharqiyah and the Dhofar. This study allowed us to isolate and identify three Leishmania strains from patients exhibiting cutaneous leishmaniasis. The typing carried out by isoenzymatic study and by molecular biology were congruent: two strains of Leishmania donovani zymodeme (Z) MON-31 isolated in the Sharqiyah and one L. tropica ZROM102 (ZMON-39 variant for 4 isoenzymes) from the Dhofar. No strain was isolated from canids. The study of sandflies identified 14 species distributed in the genera Phlebotomus, Sergentomyia and Grassomyia: Ph. papatasi, Ph. bergeroti, Ph. duboscqi, Ph. alexandri, Ph. saevus, Ph. sergenti, Se. fallax, Se. baghdadis, Se. cincta, Se. christophersi, Se. clydei, Se. tiberiadis, Se. africana, and Gr. dreyfussi. In Sharqiyah, the only candidate for the transmission of L. donovani was Ph. alexandri, but the low densities observed of this species do not argue in favor of any role. In Dhofar, Ph. sergenti is the most important proven vector of L. tropica, but Ph. saevus, a locally much more abundant species, constitutes a good candidate for transmission.

TITLE: Leishmanioses et phlébotomes au Sultanat d'Oman. ABSTRACT: Il existe peu de données sur les leishmanioses et les phlébotomes en Oman. Nous y avons mené en 1998 une étude éco-épidémiologique dans les deux principaux massifs montagneux du pays, la Sharqiyah et le Dhofar. Cette étude nous a permis d'isoler et d'identifier trois souches de Leishmania à partir de patients présentant des leishmanioses cutanées. Les typages menés par étude isoenzymatique et par biologie moléculaire ont été congruents : deux souches de Leishmania donovani ZMON-31 isolées dans la Sharqiyah et une de L. tropica ZROM102 (ZMON-39 variant pour 4 isoenzymes) originaire du Dhofar. Aucune souche n'a été isolée à partir de Canidés. L'étude des Phlébotomes a permis d'identifier 14 espèces réparties dans les genres Phlebotomus, Sergentomyia et Grassomyia : Ph. papatasi, Ph. bergeroti, Ph. duboscqi, Ph. alexandri, Ph. saevus, Ph. sergenti, Se. fallax, Se. baghdadis, Se. cincta, Se. christophersi, Se. clydei, Se. tiberiadis, Se. africana et Gr. dreyfussi. Dans la Sharqiyah, la seule espèce candidate à la transmission de L. donovani est Ph. alexandri mais les faibles densités observées de cette espèce ne plaident pas en faveur d'un quelconque rôle. Dans le Dhofar, Ph. sergenti est le principal vecteur prouvé de L. tropica mais Ph. saevus, espèce localement bien plus abondante, constitue une bonne espèce candidate à la transmission.

Two new phlebotomine sandfly species (Diptera: Psychodidae) from the Highlands of Madagascar.

The Malagasy phlebotomine sandfly fauna includes 17 species: five belong to the genus Phlebotomus, ten to the genus Sergentomyia, and two to the genus Grassomyia. The current article describes Sergentomyia brunhesi sp. nov. and Sergentomyia vistellei sp. nov. from the Malagasy Highlands. Females were described morphologically from specimens collected at Ranomafana-Ifanadiana and Andringitra. Partial molecular sequences of cyt b and 28S rDNA were retrieved for Se. vistellei sp. nov. Waiting new data, we consider these species as ungrouped Sergentomyia. Two new species of Sergentomyia are recorded at higher altitudes in Madagascar. Knowledge of local Biodiversity is increasing. New investigations have to be carried out to describe males, to understand their bionomics, and to identify other potential new species at higher altitudes.

Phlebotomine sandflies (Diptera, Psychodidae) from the Ankarana tsingy of northern Madagascar: inventory and description of new taxa.

An inventory of Phlebotomine sandflies was carried out in the Ankarana tsingy located in far northern Madagascar. A total of 723 sandflies were used for morphological, morphometric, and molecular studies (sequencing of partial cytochrome B (mtDNA) and partial 28S (rDNA)). Nine species were identified: Phlebotomus fertei, Sergentomyia anka, Se. sclerosiphon, Se. goodmani, two species of the genus Grassomyia, as well as three new species described herein: Se. volfi n. sp., Se. kaltenbachi n. sp., and Se. ozbeli n. sp. The recognition of these new species is strongly supported by molecular analyses. The first two of the new species could not be classified into any existing subgenus, therefore we proposed two new subgenera (Ranavalonomyia subg. nov., and Riouxomyia subg. nov.), with combinations as: Sergentomyia (Ranavalonomyia) volfi and Sergentomyia (Riouxomyia) kaltenbachi. Our study reveals important molecular variability in Se. anka, with the recognition of a population whose taxonomic status remains below that of species. Our research confirms the need to further study the specific diversity of Malagasy sandflies, which until the start of this millennium remained mostly unknown.

TITLE: Les phlébotomes (Diptera, Psychodidae) des tsingy d'Ankarana dans le nord de Madagascar : inventaire et description de nouveaux taxons. ABSTRACT: Un inventaire des Phlébotomes a été réalisé dans les tsingy d'Ankarana à l'extrême nord de Madagascar. Au total, 723 phlébotomes ont servi à des études morphologique, morphométrique et moléculaire (séquençage d'une partie du cytochrome B (ADNmt) et d'une partie de l'ADNr 28S). Neuf espèces ont été identifiées : Phlebotomus fertei, Sergentomyia anka, Se. sclerosiphon, Se. goodmani, deux espèces du genre Grassomyia, ainsi que trois espèces nouvelles décrites dans ce travail : Se. volfi n. sp., Se. kaltenbachi n. sp., and Se. ozbeli n. sp. L'individualisation de chacune de ces espèces nouvelles est robustement soutenue par les analyses moléculaires. Les deux premières de ces espèces nouvelles ne pouvaient pas être classées dans un sous-genre existant et nous avons proposé pour elles deux sous-genres nouveaux : Ranavalonomyia subg. nov., et Riouxomyia subg. nov, avec les combinaisons Sergentomyia (Ranavalonomyia) volfi et Sergentomyia (Riouxomyia) kaltenbachi.Notre étude révèle une variabilité moléculaire marquée chez Se. anka avec l'individualisation d'une population dont le statut taxinomique demeure populationnel. Nos travaux confirment la nécessité de poursuivre l'étude de la biodiversité des Phlébotomes qui est restée méconnue dans ce pays jusqu'au début de ce millénaire.

Molecular homogeneity in diverse geographical populations of Phlebotomus papatasi (Diptera, Psychodidae) inferred from ND4 mtDNA and ITS2 rDNA Epidemiological consequences.

An intraspecific study on Phlebotomus papatasi, the main proven vector of Leishmania major among the members of the subgenus Phlebotomus, was performed. The internal transcribed spacer 2 (ITS 2) of rDNA and the ND4 gene of mt DNA were sequenced from 26 populations from 18 countries (Albania, Algeria, Cyprus, Egypt, Greece, India, Iran, Israel, Italy, Lebanon, Morocco, Saudi Arabia, Spain, Syria, Tunisia, Turkey, Yugoslavia and Yemen), and compared. Samples also included three other species belonging to the subgenus Phlebotomus: P. duboscqi, a proven vector of L. major in the south of Sahara (three populations from Burkina Faso, Kenya and Senegal), P. bergeroti, a suspected vector of L. major (three populations from Oman Sultanate, Iran and Egypt), and one population of P. salehi from Iran. A phylogenetic study was carried out on the subgenus Phlebotomus. Our results confirm the validity of the morphologically characterized taxa. The position of P. salehi is doubtful. Variability in P. papatasi contrasts with that observed within other species having a wide distribution like P. (Paraphlebotomus) sergenti in the Old World or Lutzomyia (Lutzomyia) longipalpis in the New World. Consequently, it could be hypothesized that all populations of P. papatasi over its distribution area have similar vectorial capacities. The limits of the distribution area of L. major are correlated with the distribution of common rodents acting as hosts of the parasites.

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).

Intraspecific variation within Phlebotomus sergenti Parrot (1917) (Diptera: Psychodidae) based on mtDNA sequences in Islamic Republic of Iran.

An intraspecific study on the morphological and molecular characteristics of Phlebotomus sergenti s.l., the main vector of Leishmania tropica, was performed on 28 Iranian populations from 11 provinces and a few samples from Greece, Morocco, Lebanon, Turkey, Pakistan, and Syria. Three morphotypes were identified as A, B and C, with some intermediate forms in the samples under investigation. Based on the number of setae and the width of basal lobe of coxite, differences between A and B morphotypes were highly significant. Excluding one unusual haplotype, sequence analysis of approximately 439 bp of mtDNA (a fragment of cytochrome B gene, tRNA for serine gene, and a fragment of NADH1 gene) revealed a 6-7% genetic distance within the Iranian populations and among the specimens of other countries. Neighbor-Joining (NJ) analysis confirmed the existence of three main groups within our samples. Although there was no consistency between morphotypes and genotypes, but an interrelationship was found between morphometry and morphotypes. Morphotype A, which was considered as P. sergenti sergenti, was the most prevalent in collection sites. Morphotype B, which was identified as Phlebotomus sergenti similis, is the first record of this subspecies in Iran, and was found to be sympatric with other morphotypes. Morphotype C had an elongated style in comparison with P. sergenti sergenti. Molecular database showed three main genetic structures. This is the first combined morphological and molecular studies on P. sergenti s.l. in Iran.

Phlebotomus (Paraphlebotomus) chabaudi and Phlebotomus riouxi: closely related species or synonyms?

Phlebotomus riouxi Depaquit, Killick-Kendrick & Léger 1998 was described as a species closely related to Phlebotomus chabaudi Croset, Abonnenc & Rioux 1970, differing mainly by the size and number of setae of the coxite basal lobe. Molecular studies carried out on several populations from Algeria and Tunisia and based on mitochondrial genes cytochrome b (Cytb) and cytochrome oxidase I (COI) supported the typological validity of these two species. Recently, specimens from a single population in southern Tunisia were morphologically identified as Ph. riouxi, Ph. chabaudi and intermediates, but were clustered in the same clade according to their Cytb and nuclear gene elongation factor-1 α (EF-1α) sequences. These species were thus synonymized. To further explore this synonymy, we carried out a molecular study on specimens from Algeria and Tunisia using the same molecular markers and a part of 28S rDNA. We did not find any morphologically intermediate specimens in our sampling. We highlighted differences between the genetic divergence rates within and between the two species for the three markers and we identified new haplotypes. The sequence analysis did not reveal any signature of introgression in allopatric nor in sympatric populations such as in the Ghomrassen population. Phylogenetic analyses based on our specimens revealed that the two main clades are Ph. chabaudi and Ph. riouxi, in agreement with the morphological identification. These results support the validity of Ph. riouxi and Ph. chabaudi as typological species.

An illustrated guide for characters and terminology used in descriptions of Phlebotominae (Diptera, Psychodidae).

Phlebotomine (Diptera, Psychodidae, Phlebotominae) taxonomy has been studied extensively, primarily due to the role of these flies as vectors of various parasites, including species of Leishmania, Bartonella and arboviruses that cause diseases in humans and other vertebrates. We present some topics discussed at a round-table on phlebotomine taxonomy held at the Ninth International Symposium on Phlebotomine Sandflies (ISOPS IX) in Reims, France, in June 2016. To date, approximately one thousand phlebotomine species have been described worldwide, although in varying languages and mostly without standardization of characters and terminology. In the interest of standardization, we list the characters that should minimally be considered in the description of new phlebotomine taxa as well as annotated illustrations of several characters. For these characters, multiple illustrations are provided to show some of the variations. The preferred terms for all pertinent characters are listed as well as their synonyms in English, Portuguese, and French. Finally, we offer an updated list of abbreviations to be used for generic and subgeneric names.

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.

Idiophlebotomus padillarum n. sp. (Diptera Psychodidae) a new sand fly species from Palawan (Philippines).

A new species of Phlebotomine sandfly is described from a cave located in Palawan, an island situated on the West of the Wallace-Huxley line in the Oriento-Australian generalized area. It has been classified among the Idiophlebotomus according to morphological characters. Male and female have been recorded in the same species on the basis of the identity of their mtDNA cytochrome b sequences. Descriptions are illustrated by drawings using a camera lucida.

Paraphyly of the subgenus Sintonius (Diptera, Psychodidae, Sergentomyia): status of the Malagasy species. Creation of a new subgenus and description of a new species.

During an inventory of Phlebotomine sand flies carried out in Madagascar, we have identified some specimens showing morphological characters related to the subgenus Sintonius of the genus Sergentomyia. We started a molecular study based on cytochrome b mtDNA and on D1-D2 and D8 domains of the rDNA. The sampling includes all the Sergentomyia species available and also S. (Sergentomyia) schwetzi, S. (Parrotomyia) magna, and the following species belonging to the subgenus Sintonius: S. clydei, S. christophersi, S. affinis vorax, S. adleri and S. meilloni. The Sintonius subgenus (sensu Theodor) is paraphyletic. The Malagasy specimens morphologically Sintonius-like are never clustered with the continental Sintonius. We propose a new subgenus to include them: Trouilletomyia subg. nov. Due to the lack of mesanepisternal setae, the species huberti is removed from the genus Phlebotomus and we propose here a new combination: Sergentomyia huberti comb. nov. The male of S. huberti is pinpointed and described for the first time. Lastly, a new species for Science is described on one female: Sergentomyia (Trouilletomyia) boironis n. sp.

Phlebotomine sand flies from Madagascar (Diptera: Psychodidae). VII. An identification key for Phlebotomus with the description of Phlebotomus (Anaphlebotomus) vaomalalae n. sp.

An identification key of the Phlebotomus in Madagascar is proposed as well as the description of the male and female Phlebotomus (Anaphlebotomus) vaomalalae n. sp. from Mikea Forest in the south-west of Madagascar. The assignation of this new species to the genus Phlebotomus is based on the presence of mesanepisternal setae. Its inclusion in the subgenus Anaphlebotomus is based on the males on the presence of four spines on the style, the lack of a coxite basal process and the existence of a bifurcated paramere. The female has cibarial and pharyngeal armature and spermathecal architecture similar to Phlebotomus fertei and Phlebotomus berentiensis, two other Malagasy species which belong to Anaphlebotomus. Male and female are held to belong to the same species because of their morphological characters, the homology (100%) of their partial cytochrome b mtDNA sequences and their capture in the same trap. P. vaomalalae n. sp. is a small species compared to the other Phlebotomus species of Madagascar. The cibarium of the male and the female of P. vaomalalae n. sp. is armed with teeth, like those of other Malagasy Phlebotomus. However, it differs in the arrangement and shape of the respective teeth and denticles. The male of P. vaomalalae n. sp. looks like that of P. fontenillei due to its tuft of coxal setae (lacking in P. berentiensis and P. fertei) but differs from this species by the location of this tuft. As P. fertei and P. berentiensis, there is no spermathecal common duct in P. vaomalalae n. sp.

A taxonomic study of Phlebotomus (Larroussius) perfiliewi s. l.

Phlebotomus (Larroussius) perfiliewi is one of the main vectors of Leishmania infantum in the Mediterranean basin and in Central Asia. Its taxonomic status remains doubtful. It usually includes three taxa of specific or subspecific value, depending on the authors: P. perfiliewiParrot, 1930 s. st. in North Africa, Western Europe and the Balkans, Crimea; Phlebotomus galilaeusTheodor, 1958 in Israel/Palestine and Cyprus and Phlebotomus transcaucasicusPerfiliev, 1937 in Caucasus and Central Asia. While the identification of the females of P. perfiliewi s. l. is relatively easy, based on the particular morphology of their spermathecal ducts, the specific identification of the females of Phlebotomus perfiliewi perfiliewi, Phlebotomus perfiliewi galilaeus and Phlebotomus perfiliewi transcaucasicus is impossible. The differentiation of males, based mainly on characters on aedeagus, is also difficult. The goal of the present study is to carry out a taxonomic update based on a morphological study of specimens coming from various populations from Asia, Europe and North Africa coupled with their molecular analysis.

Description of the sandfly species Chinius samarensis n. sp. (Psychodidae; Diptera) from the Philippines.

The authors have described Chinius samarensis n. sp. (Diptera; Psychodidae) from the Philippines (Island of Samar). This is the fourth known species belonging to the Asiatic genus Chinius Leng, 1987. Like the other Chinius, C. samarensis n. sp. is a cavernicolous species. The genital ducts of the male are four times shorter than the spermathecal ducts. However, the authors think that male and female belong to the same species due to their similar cytochrome b mt DNA sequences. A differential diagnosis with the other species belonging to the genus Chinius is provided.

Phlebotomus (Euphlebotomus) barguesae n. sp. from Thailand (Diptera - Psychodidae).

BACKGROUND: A few studies have been carried out on the Phlebotomine sandflies from Thailand. Within the Phlebotomine sandflies, the genus Phlebotomus Rondani & Berté, 1840 contains the vectors of leishmaniases in Europe, Africa and Asia. It includes several subgenera. Among them the subgenus Euphlebotomus Theodor, 1948 contains at the present time 12 taxa. The type-species of this subgenus is P. argentipes Annandale & Brunetti, 1908, the vector of Leishmania donovani (Laveran & Mesnil, 1903) in India. RESULTS: A new species of sandfly, P. barguesae n. sp. is described from limestone caves in Thailand. The male-female gathering in the same species is based on ecological, morphological and molecular criteria (homology of mtDNA cytochrome c oxidase I sequences). The inclusion of P. barguesae n. sp. in the subgenus Euphlebotomus is justified on the basis of characters of the male genitalia (five spines on the style, bifurcated paramere, and no basal lobe on the coxite) and of female pharyngeal armature (two kinds of teeth). It well differenciated from another sympatric species: P. mascomai. CONCLUSION: The new species described in the present study has smooth spermathecae. This original morphology opens a discussion on the heterogeneity of this subgenus.

Trichobilharzia spp. in natural conditions in Annecy Lake, France.

Annecy Lake is a well-known focus of human cercarial dermatitis in France. Identification of the parasites, however, was not performed in the past. Previous studies suspected two species, Trichobilharzia franki and Trichobilharzia regenti, based on the presence of parasites in mallards and/or morphological identification of snails emitting ocellate furcocercariae. Following a standardized molecular approach, we studied snails and furcocercariae and compared their haplotypes with those deposited in GenBank. The selected markers were the second internal transcribed spacer ITS-2 for the snails and ITS-2 and D2 domain of the ribosomal DNA for the parasites. Our results confirm the presence of T. franki and T. regenti and two probable new species that could be potential agents of cercarial dermatitis. All the snails emitting the ocellate furcocercariae belong to the same species identified as Radix peregra (=Radix ovata = Radix balthica). Parasite-host relationships between species of the genus Trichobilharzia and snails of the genus Radix do not seem to be as specific as supposed previously.

Phlebotomus (Euphlebotomus) mascomai n. sp. (Diptera-Psychodidae).

A new species of sandfly is described from limestone caves in Thailand. The inclusion of this species in the subgenus Euphlebotomus is justified on the basis of characters of the male genitalia (paramere, basal lobe). The male-female gathering in the same taxon is based on ecological (cavernicolous species), morphological (length of male genital filaments and female spermathecal ducts) and molecular (homology of cytochrome b mt DNA sequences) criteria. A differential diagnosis between Phlebotomus mascomai n. sp. and P. argentipes Annandale & Brunetti, the vector of Leishmania donovani (Laveran & Mesnil) in India, is proposed based on several morphological characters like antennal formula and genitalia.