Significance of eggshell morphology as an additional tool to distinguish species of sand flies (Diptera: Psychodidae: Phlebotominae).

Morphological characteristics of eggshells are important in sand fly ootaxonomy. In this study, eggshells from Phlebotomus stantoni Newstead, Sergentomyia khawi (Raynal), and Grassomyia indica (Theodor) sand flies collected in Chiang Mai province, Thailand were examined and characterized using light microscopy (LM) and scanning electron microscopy (SEM). Then, eggshell morphology of these three species was described for the first time. Each gravid female was forced to lay eggs by decapitation and the eggs were collected for SEM analysis. Egg laying females were identified by morphological examination and molecular typing using cytochrome b (Cytb) as a molecular marker. The chorionic sculpturing of Ph. stantoni eggs combines two patterns on the same egg: unconnected parallel ridges and reticular patterns. Sergentomyia khawi and Gr. indica have similar chorionic polygonal patterns, but their exochorionic morphology and aeropylar area are different. Results indicate that eggshell morphological characteristics such as chorionic pattern, exochorionic morphology, inter-ridge/boundary area, aeropylar area (including the number of aeropyles) and basal layer, can be useful to develop morphological identification keys of eggs. These can serve as an additional tool to distinguish species of sand flies. In addition, the chorionic sculpturing of the eggs of the three species of sand flies observed by LM is useful for species identification in gravid females with spermathecae obscured by eggs.

Species identification and phylogenetic analysis of Leishmania isolated from patients, vectors and hares in the Xinjiang Autonomous Region, The People's Republic of China.

BACKGROUND: Visceral leishmaniasis (VL) has been declared as one of the six major tropical diseases by the World Health Organization. This disease has been successfully controlled in China, except for some areas in the western region, such as the Xinjiang Autonomous Region, where both anthroponotic VL (AVL) and desert type zoonotic VL (DT-ZVL) remain endemic with sporadic epidemics. METHODOLOGY/PRINCIPAL FINDINGS: Here, an eleven-year survey (2004-2014) of Leishmania species, encompassing both VL types isolated from patients, sand-fly vectors and Tarim hares (Lepus yarkandensis) from the Xinjiang Autonomous Region was conducted, with a special emphasis on the hares as a potential reservoir animal for DT-ZVL. Key diagnostic genes, ITS1, hsp70 and nagt (encoding N-acetylglucosamine-1-phosphate transferase) were used for phylogenetic analyses, placing all Xinjiang isolates into one clade of the L. donovani complex. Unexpectedly, AVL isolates were found to be closely related to L. infantum, while DT-ZVL isolates were closer to L. donovani. Unrooted parsimony networks of haplotypes for these isolates also revealed their relationship. CONCLUSIONS/SIGNIFICANCE: The above analyses of the DT-ZVL isolates suggested their geographic isolation and independent evolution. The sequence identity of isolates from patients, vectors and the Tarim hares in a single DT-ZVL site provides strong evidence in support of this species as an animal reservoir.

Spatial and Temporal Variability of Visceral Leishmaniasis in Colombia, 2007 to 2018.

Visceral leishmaniasis (VL) is a neglected tropical disease associated with poverty and is endemic in 56 countries worldwide. Brazil, Venezuela, and Colombia are the most affected countries in South America. In Colombia, the National Public Health Surveillance System (SIVIGILA) consolidates epidemiological information and monitors all VL cases nationwide. However, to date, no studies have investigated the occurrence of VL in Colombia using metadata analysis. We studied the demographic data, the spatial and temporal distribution of VL cases, and the association with vector distribution of Leishmania species in Colombia from 2007 to 2018. We found 306 VL cases reported to SIVIGILA for this period, with a coverage of 25.5 cases/year, and a mortality of 2.28% (seven deaths). The highest number of confirmed cases (N = 52) occurred in 2007; the lowest (N = 9) occurred in 2012. The cases were reported mainly in children (< 7 years) affiliated with the subsidized health regimen. Regarding the geographic distribution, the cases were reported by 42 municipalities distributed in 10 departments. The occurrence of VL cases toward the northeast of Colombia, and the distribution of vectors, such as Lutzomyia longipalpis and Lu. evansi, may be changing the panorama of VL in the country. We conclude that VL, mainly in recent years, shows a temporal and spatial variability associated with the occurrence of cases in new settings. Our findings increase our understanding and knowledge of this disease, and suggest the need to monitor and prioritize areas with changes in geographic expansion to improve prevention and control actions in the country.

Phlebotomine sand fly survey in the Republic of Moldova: species composition, distribution and host preferences.

BACKGROUND: Phlebotomine sand flies (Diptera: Psychodiae) in the Republic of Moldova have been understudied for decades. Our study provides a first update on their occurrence, species composition and bloodmeal sources after 50 years. METHODS: During 5 seasons (2013-2017), 58 localities from 20 regions were surveyed for presence of sand flies using CDC light traps and manual aspirators. Species identification was done by a combination of morphological and molecular approaches (DNA barcoding, MALDI-TOF MS protein profiling). In engorged females, host blood was identified by three molecular techniques (RFLP, cytb sequencing and MALDI-TOF peptide mass mapping). Population structure of most abundant species was studied by cox1 haplotyping; phylogenetic analyses of ITS2 and cox1 genetic markers were used to resolve relationships of other detected species. RESULTS: In total, 793 sand flies were collected at 30 (51.7%) localities from 12 regions of Moldova. Three species were identified by an integrative morphological and molecular approach: Phlebotomus papatasi, P. perfiliewi and Phlebotomus sp. (Adlerius), the first being the most abundant and widespread, markedly anthropophilic based on bloodmeal analyses, occurring also indoors and showing low population structure with only five haplotypes of cox1 detected. Distinct morphological and molecular characters of Phlebotomus sp. (Adlerius) specimens suggest the presence of a yet undescribed species. CONCLUSIONS: Our study revealed the presence of stable sand fly populations of three species in Moldova that represent a biting nuisance as well as a potential threat of pathogen transmission and shall be further studied.

Efficacy of afoxolaner (NexGard®) in preventing the transmission of Leishmania infantum and Dirofilaria immitis to sheltered dogs in a highly endemic area.

BACKGROUND: Leishmania infantum and Dirofilaria immitis are among the most important canine vector-borne pathogens (CVBPs) of zoonotic concern in Europe. In endemic areas for both of these CVBPs, the use of systemic ectoparasiticides, such as afoxolaner (NexGard®; Boehringer Ingelheim Animal Health), may have the potential for controlling these infections. The aim of this study was to assess, for the first time, the insecticidal efficacy of NexGard® in decreasing the transmission of D. immitis and L. infantum to sheltered dogs living in a hyperendemic area, compared to the year before treatment, as well as its impact on the abundance of mosquito and sand fly populations. METHODS: All dogs (n = 179) enrolled in the study were divided into two groups based on their infection status at enrollment: a non-infected group (G1) and an infected group (G2; infected with D. immitis, L. infantum or both). The study was conducted from March 2020 to March 2021. In order to exclude all animals infected with L. infantum and D. immitis before March 2020 (sampling time: T0), dogs in G1 were sampled in June (T1; i.e. T0 + 90 days) and in October 2020 (T2; i.e. T0 + 210 days). From March to September 2020, all animals (G1 and G2) were weighed and treated monthly with NexGard®. Animals in G1 were tested for the last time in March 2021 (T3; i.e. T0 + 330 days) for assessing post-treatment incidence rate of infection and prevention efficacy. RESULTS: The post-treatment incidence of D. immitis was 3.7% (1/27; 95% confidence interval [CI]: 0.2-18.1) and that of L. infantum was 3.6% (3/83; 95% CI: 1.0-10.1). Considering the annual incidence in 2019 and 2020, the protective efficacy against D. immitis and L. infantum infections was 94.2 and 64%, respectively. Of the female mosquitoes collected (n = 146), only one pool out of 50 tested positive for D. immitis DNA, whereas out of 1252 female Sergentomya minuta specimens collected, only four tested positive for L. infantum (0.3%). CONCLUSIONS: Afoxolaner is efficacious in decreasing the rate of transmission of both D. immitis and L. infantum; however, comparison of the pre- and post-treatment period demonstrated that there was a significant difference only in the seasonal incidences of D. immitis infection. Preventive measures are recommended throughout the year in endemic areas to reduce the risk of pathogen transmission to animals and humans.

Multiple evolutionary lineages for the main vector of Leishmania guyanensis, Lutzomyia umbratilis (Diptera: Psychodidae), in the Brazilian Amazon.

Lutzomyia umbratilis is the main vector of Leishmania guyanensis in the Brazilian Amazon and in neighboring countries. Previous biological and molecular investigations have revealed significant differences between L. umbratilis populations from the central Brazilian Amazon region. Here, a phylogeographic survey of L. umbratilis populations collected from nine localities in the Brazilian Amazon was conducted using two mitochondrial genes. Statistical analyses focused on population genetics, phylogenetic relationships and species delimitations. COI genetic diversity was very high, whereas Cytb diversity was moderate. COI genealogical haplotypes, population structure and phylogenetic analyses identified a deep genetic differentiation and three main genetic groups. Cytb showed a shallower genetic structure, two main haplogroups and poorly resolved phylogenetic trees. These findings, allied to absence of isolation by distance, support the hypothesis that the Amazon and Negro Rivers and interfluves are the main evolutionary forces driving L. umbratilis diversification. The main three genetic groups observed represent three evolutionary lineages, possibly species. The first lineage occurs north of the Amazon River and east of Negro River, where Le. guyanensis transmission is intense, implying that L. umbratilis is an important vector there. The second lineage is in the interfluve between north of Amazon River and west of Negro River, an area reported to be free of Le. guyanensis transmission. The third lineage, first recorded in this study, is in the interfluve between south of Amazonas River and west of Madeira River, and its involvement in the transmission of this parasite remains to be elucidated.

Two new Palaearctic species of moth flies (Diptera, Psychodidae, Psychodinae) from the Caucasus Mts.

Psychodid specimens were sorted from samples collected during fieldwork in areas in and around the Caucasus Mountains in 2019. Thornburghiella montana sp. nov. (from Georgia) and Pericoma inopinata sp. nov. (from Azerbaijan and Georgia) are described. Differential diagnoses are given and diagnostic characters illustrated.

New species of Boreofairchildia Wagner amp; Stuckenberg and Laurenceomyia Wagner amp; Stuckenberg (Diptera: Psychodidae: Bruchomyiinae) from Brazil.

Two new species of Bruchomyiinae Macquart from Brazil are described and illustrated. Laurenceomyia peixotoi Santos, Brazil Pinto sp. nov. to Amazon Forest and Boreofairchildia alexanderi Santos, Brazil Pinto sp. nov. to Atlantic Forest.

A follow-up study (2007-2018) on American Tegumentary Leishmaniasis in the municipality of Caratinga, Minas Gerais State, Brazil: Spatial analyses and sand fly collection.

BACKGROUND: The municipality of Caratinga is an important endemic area for American Tegumentary Leishmaniasis (ATL) and no epidemiological studies were performed during the past two decades. Here, we analyzed the epidemiological situation and the geographical distribution of ATL cases in the municipality of Caratinga from 2007 to 2018 using geographic information systems (GIS). Also, we evaluated the impact of several demographic parameters in ATL distribution and the sand flies incriminated in its transmission. METHODS: All demographic information (gender, age, educational level, clinical form, diagnostic criteria and case evolution) used in this study was retrieved from the public health archives and confirmed in the State Health Services databases. All cases were analyzed using GIS software based on ATL distribution. Also, non-systematic sand fly collections and molecular detection of Leishmania were performed in the hotspots. RESULTS AND CONCLUSIONS: During the period, ATL cases continued and increased especially in the past years (2016-2018). Hotspots included urban Caratinga areas and the districts of Patrocínio de Caratinga and Sapucaia. The species Nyssomyia whitmani, Nyssomyia intermedia, Migonemyia migonei and Evandromyia cortelezzii complex were captured. However, ITS1-PCR did not detect Leishmania DNA in those insects. Based on our analyses, urbanization of ATL in Caratinga has occurred in the past years. Due to the increase in the number of cases and vectors presence, it is recommended that health authorities focus on control measures in the most affected areas (Patrocínio of Caratinga and Sapucaia districts and urban Caratinga).

Sand Fly Fauna, Spatial Distribution of Lutzomyia longipalpis (Diptera: Psychodidae), and Climate Factors in Dourados, Brazil.

Studies of the geographic distribution of sand flies and the factors associated with their occurrence are necessary to understand the risk of leishmaniasis transmission. The objective of this study was to characterize the sand fly fauna, particularly the spatial distribution of Lutzomyia longipalpis (Lutz & Neiva), and correlate these with climate factors in the Dourados municipality, Brazil. The collection of sand flies was carried out with CDC Light Traps over two periods: at six sites for three consecutive nights each month from August 2012 to July 2013; and at four other sites for two consecutive nights each month from April 2017 to February 2018. We collected 591 sand flies in the first period and 121 in the second period for a total of 712 sand flies; 697 of the total collected were Lu. longipalpis. The minimum and maximum sand fly infestation rate (sites with vector presence) was 11.1% and 83.33% in the first period, and 0% and 50.0% in the second period. No sand flies with Leishmania were identified via PCR. Lu. longipalpis presented an aggregate disposition with excellent adjustment. Rainfall and relative humidity were the abiotic factors that influenced the vector infestation level. The aggregate distribution for this species was predicted by the environmental factors that favor the proliferation of Lu. longipalpis. The results of this study should assist in devising measures to control sand flies in Dourados, Brazil.

Molecular detection of Leishmania donovani, Leishmania major, and Trypanosoma species in Sergentomyia squamipleuris sand flies from a visceral leishmaniasis focus in Merti sub-County, eastern Kenya.

BACKGROUND: Visceral leishmaniasis (VL) and zoonotic cutaneous leishmaniasis (ZCL) are of public health concern in Merti sub-County, Kenya, but epidemiological data on transmission, vector abundance, distribution, and reservoir hosts remain limited. To better understand the disease and inform control measures to reduce transmission, we investigated the abundance and distribution of sand fly species responsible for Leishmania transmission in the sub-County and their blood-meal hosts. METHODS: We conducted an entomological survey in five villages with reported cases of VL in Merti sub-County, Kenya, using CDC miniature light traps and castor oil sticky papers. Sand flies were dissected and identified to the species level using standard taxonomic keys and PCR analysis of the cytochrome c oxidase subunit 1 (cox1) gene. Leishmania parasites were detected and identified by PCR and sequencing of internal transcribed spacer 1 (ITS1) genes. Blood-meal sources of engorged females were identified by high-resolution melting analysis of vertebrate cytochrome b (cyt-b) gene PCR products. RESULTS: We sampled 526 sand flies consisting of 8 species, Phlebotomus orientalis (1.52%; n = 8), and 7 Sergentomyia spp. Sergentomyia squamipleuris was the most abundant sand fly species (78.71%; n = 414) followed by Sergentomyia clydei (10.46%; n = 55). Leishmania major, Leishmania donovani, and Trypanosoma DNA were detected in S. squamipleuris specimens. Humans were the main sources of sand fly blood meals. However, we also detected mixed blood meals; one S. squamipleuris specimen had fed on both human and mouse (Mus musculus) blood, while two Ph. orientalis specimens fed on human, hyrax (Procavia capensis), and mouse (Mus musculus) blood. CONCLUSIONS: Our findings implicate the potential involvement of S. squamipleuris in the transmission of Leishmania and question the dogma that human leishmaniases in the Old World are exclusively transmitted by sand flies of the Phlebotomus genus. The presence of Trypanosoma spp. may indicate mechanical transmission, whose efficiency should be investigated. Host preference analysis revealed the possibility of zoonotic transmission of leishmaniasis and other pathogens in the sub-County. Leishmania major and L. donovani are known to cause ZCL and VL, respectively. However, the reservoir status of the parasites is not uniform. Further studies are needed to determine the reservoir hosts of Leishmania spp. in the area.

Potential vectors of Leishmania spp. in an Atlantic Forest conservation unit in northeastern Brazil under anthropic pressure.

BACKGROUND: Phlebotomines are a group of insects which include vectors of the Leishmania parasites that cause visceral leishmaniasis (VL) and cutaneous leishmaniasis (CL), diseases primarily affecting populations of low socioeconomic status. VL in Brazil is caused by Leishmania infantum, with transmission mainly attributed to Lutzomyia longipalpis, a species complex of sand fly, and is concentrated mainly in the northeastern part of the country. CL is distributed worldwide and occurs in five regions of Brazil, at a higher incidence in the north and northeast regions, with etiological agents, vectors, reservoirs and epidemiological patterns that differ from VL. The aim of this study was to determine the composition, distribution and ecological relationships of phlebotomine species in an Atlantic Forest conservation unit and nearby residential area in northeastern Brazil. METHODS: Centers for Disease Control and Shannon traps were used for collections, the former at six points inside the forest and in the peridomestic environment of surrounding residences, three times per month for 36 months, and the latter in a forest area, once a month for 3 months. The phlebotomines identified were compared with climate data using simple linear correlation, Pearson's correlation coefficient and cross-correlation. The estimate of ecological parameters was calculated according to the Shannon-Wiener diversity index, standardized index of species abundance and the dominance index. RESULTS: A total of 75,499 phlebotomines belonging to 11 species were captured in the CDC traps, the most abundant being Evandromyia walkeri, Psychodopygus wellcomei and Lu. longipalpis. Evandromyia walkeri abundance was most influenced by temperature at collection time and during the months preceding collection and rainfall during the months preceding collection. Psychodopygus wellcomei abundance was most affected by rainfall and relative humidity during the collection month and the month immediately preceding collection time. Lutzomyia longipalpis abundance showed a correlation with temperature and the rainfall during the months preceding collection time. The Shannon trap contained a total of 3914 phlebotomines from these different species. Psychodopygus wellcomei, accounting for 91.93% of the total, was anthropophilic and active mainly at night. CONCLUSIONS: Most of the species collected in the traps were seasonal and exhibited changes in their composition and population dynamics associated with local adaptions. The presence of vectors Ps. wellcomei and Lu. longipalpis underscore the epidemiological importance of these phlebotomines in the conservation unit and surrounding anthropized areas. Neighboring residential areas should be permanently monitored to prevent VL or CL transmission and outbreaks.

Phlebotomus (Adlerius) simici NITZULESCU, 1931: first record in Austria and phylogenetic relationship with other Adlerius species.

BACKGROUND: Phlebotomine sand flies are the principal vectors of Leishmania spp. (Kinetoplastida: Trypanosomatidae). Information on sand flies in Central Europe is scarce and, to date, in Austria, only Phlebotomus mascittii has been recorded. In 2018 and 2019, entomological surveys were conducted in Austria with the aim to further clarify sand fly distribution and species composition. RESULTS: In 2019, a Ph. simici specimen was trapped in Austria for the first time. Analyses of two commonly used marker genes, cytochrome c oxidase I (coxI) and cytochrome b (cytb), revealed high sequence identity with Ph. simici specimens from North Macedonia and Greece. Phylogenetic analyses showed high intraspecific distances within Ph. simici, thereby dividing this species into three lineages: one each from Europe, Turkey and Israel. Low interspecific distances between Ph. simici, Ph. brevis and an as yet unidentified Adlerius sp. from Turkey and Armenia highlight how challenging molecular identification within the Adlerius complex can be, even when standard marker genes are applied. CONCLUSION: To our knowledge, this study reports the first finding of Ph. simici in Austria, representing the northernmost recording of this species to date. Moreover, it reveals valuable insights into the phylogenetic relationships among species within the subgenus Adlerius. Phlebotomus simici is a suspected vector of L. infantum and therefore of medical and veterinary importance. Potential sand fly expansion in Central Europe due to climatic change and the increasing import of Leishmania-infected dogs from endemic areas support the need for further studies on sand fly distribution in Austria and Central Europe in general.

Detection of Pintomyia fischeri (Diptera: Psychodidae) With Leishmania infantum (Trypanosomatida: Trypanosomatidae) Promastigotes in a Focus of Visceral Leishmaniasis in Brazil.

Visceral leishmaniasis is spreading in Brazil where the main vector of its agent, Leishmania infantum Nicolle, 1908, is the Lutzomyia longipalpis (Lutz & Neiva, 1912) species complex (Diptera: Psychodidae: Phlebotominae), on which many of the activities of the visceral leishmaniasis surveillance program are based. However, there are areas where canine, and/or human cases have been occurring without the presence of this species complex as in the western part of the Greater São Paulo Metropolitan region, where Embu das Artes municipality is situated. In this area, Pintomyia fischeri (Pinto, 1926) has been implicated as potential vector of Le. infantum but so far its natural infection with this parasite has not yet been ascertained. Therefore, the present study sought to investigate the natural infection in sand flies of a CVL focus in Embu das Artes. The sand fly collections were undertaken with Shannon and CDC traps, monthly, between 1800 and 2100 hours from November 2018 to October 2019, inclusive. A total of 951 sand flies (457 males and 494 females), belonging to 10 species, were captured. Pintomyia fischeri was the predominant species (89.5%); of which 426 females were dissected and one of them (0.23%) was found to be harboring flagellates in its midgut. A sample of these flagellates was isolated in culture and characterized by a 234 base pair fragment of Leishmania heat-shock protein 70 gene (hsp70) and restriction fragment length polymorphism with Hae III restriction enzyme as Le. infantum. This finding reinforces previous evidence of Pi. fischeri as a vector of Le. infantum in foci of visceral leishmaniasis and highlights the importance of vector surveillance in areas where this species occurs.

Redescription of Two Psathyromyia Species (Diptera: Psychodidae), Including Description of the Female of Psathyromyia pradobarrientosi Using Molecular and Morphological Approaches.

The taxonomic identity of two species of sand flies, Psathyromyia pradobarrientosi (Le Pont, Matias, Martinez & Dujardin, 2004) and Psathyromyia runoides (Fairchild & Hertig, 1953) (Diptera, Psychodidae), was evaluated morphologically and molecularly based upon specimens collected in Brazilian states. The morphological component compared collected specimens with paratypes of Pa. runoides and Pa. pradobarrientosi and their descriptions. Phylogenetic analysis of coI sequences of Pa. pradobarrientosi showed a well-supported group distinct from Pa. runoides. Morphologically, Psathyromyia runoides and Pa. pradobarrientosi males are distinguished by characteristics of the aedeagal ducts and parameral sheath in the genitalia; females are distinguished by the number and shape of the teeth in the cibarium and by the shape of the spermathecae. Given the morphological similarity between the males and the absence of the description of the female of Pa. pradobarrientosi, it is possible that specimens previously identified as Pa. runoides in Brazil are in fact Pa. pradobarrientosi.

TRsandflies: A Web-Based Software for the Morphometric Identification of Sand Flies in Turkey.

Sand flies are vector of several diseases, mostly cutaneous and visceral leishmaniasis (CL and VL). Also, 29 sand fly species have been identified in previous fauna studies carried out in 40 provinces of Turkey. Totally, 24 sand flies species belonging to Phlebotomus (Ph.) (Diptera: Psychodidae) genus have been proven or reported as possible vector species. This study aimed to develop a new software which could contribute to researchers' decision making about the identification of sand flies with obtained data from entomological surveys conducted before in Turkey. Developed software called TRsandflies included 35 textbox created with parameters obtained from caught sand flies specimens by the above-mentioned surveys. It also contained 130 photos and distribution maps related to 24 sand flies species. In addition, C# language and MYSQL database were used in the program. TRsandflies had three different forms (pages) allowing the user to compare the specimens and known species. In the species identification trials with three repetitions carried out in the program, except for the specimens belonging to the Transphlebotomus Artemiev & Neronov, 1984 subgenus, morphometric data of all previously collected sand fly species specimens were included. The process of running the morphometric measurement results of predetermined specimens in the program provided us with an accurate prediction rate of 86.66% in male specimens and 71.66% in female specimens. We concluded that the web-based software developed could play an important role in reducing the rate of possible errors that might be encountered by conventional identification methods.

Characterization of the sand fly fauna in Barbalha, one of the municipalities with the highest leishmaniasis rates in Brazil.

Leishmaniases are a complex of sand fly-borne diseases that are considered a public health issue in several countries. Brazil presents high leishmaniases rates. The South of Ceará State, known as Cariri region, shows worrying statistics mainly on American tegumentary leishmaniasis. In Barbalha, which is one of the municipalities in this region, there is still a lack of studies regarding the local phlebotomine (Diptera: Psychodidae) fauna in order to help clarify the high rates. This study aimed to characterize such fauna by capturing sand flies with light traps during a four-year period. A total of 3730 sand flies were captured, of which 37.8% were females. Fourteen species were found: 13 of the Lutzomyia genus and one of the Brumptomyia genus. Of the Lutzomyia species, four were proven and five had potential involvement in leishmaniasis transmission. Lutzomyia longipalpis was the most common species (66.97%). This predominance, especially in the urban area, indicates its epidemiological importance and adaptation to environmental conditions modified by human activity. In fact, further studies are still required to accurately determine the behavioral features of these vectors in order to guide public health measurements towards its control and prevention.

Characterization of a Novel Mitovirus of the Sand Fly Lutzomyia longipalpis Using Genomic and Virus-Host Interaction Signatures.

Hematophagous insects act as the major reservoirs of infectious agents due to their intimate contact with a large variety of vertebrate hosts. Lutzomyia longipalpis is the main vector of Leishmania chagasi in the New World, but its role as a host of viruses is poorly understood. In this work, Lu. longipalpis RNA libraries were subjected to progressive assembly using viral profile HMMs as seeds. A sequence phylogenetically related to fungal viruses of the genus Mitovirus was identified and this novel virus was named Lul-MV-1. The 2697-base genome presents a single gene coding for an RNA-directed RNA polymerase with an organellar genetic code. To determine the possible host of Lul-MV-1, we analyzed the molecular characteristics of the viral genome. Dinucleotide composition and codon usage showed profiles similar to mitochondrial DNA of invertebrate hosts. Also, the virus-derived small RNA profile was consistent with the activation of the siRNA pathway, with size distribution and 5' base enrichment analogous to those observed in viruses of sand flies, reinforcing Lu. longipalpis as a putative host. Finally, RT-PCR of different insect pools and sequences of public Lu. longipalpis RNA libraries confirmed the high prevalence of Lul-MV-1. This is the first report of a mitovirus infecting an insect host.

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.

Sand fly fauna of Crete and the description of Phlebotomus (Adlerius) creticus n. sp. (Diptera: Psychodidae).

BACKGROUND: The Greek island of Crete is endemic for both visceral leishmaniasis (VL) and recently increasing cutaneous leishmaniasis (CL). This study summarizes published data on the sand fly fauna of Crete, the results of new sand fly samplings and the description of a new sand fly species. METHODS: All published and recent samplings were carried out using CDC light traps, sticky traps or mouth aspirators. The specific status of Phlebotomus (Adlerius) creticus n. sp., was assessed by morphological analysis, cytochrome b (cytb) sequencing and MALDI-TOF protein profiling. RESULTS: Published data revealed the presence of 10 Phlebotomus spp. and 2 Sergentomyia spp. During presented field work, 608 specimens of 8 species of Phlebotomus and one species of Sergentomyia were collected. Both published data and present samplings revealed that the two most common and abundant species were Phlebotomus neglectus, a proven vector of Leishmania infantum causing VL, and Ph. similis, a suspected vector of L. tropica causing CL. In addition, the field surveys revealed the presence of a new species, Ph. (Adlerius) creticus n. sp. CONCLUSIONS: The identification of the newly described species is based on both molecular and morphological criteria, showing distinct characters of the male genitalia that differentiate it from related species of the subgenus Adlerius as well as species-specific sequence of cytb and protein spectra generated by MALDI-TOF mass spectrometry.