Descriptive and Geometric Morphometry of the Wings of Phlebotomus sergenti Populations in Central Morocco

Background: Phlebotomus sergenti, the proven vector of Leishmania tropica, the causative agent of anthroponotic cu­taneous leishmaniasis, is widely distributed in Morocco. Previous works using molecular markers (Internal Transcribed Spacer 2 rDNA and Cytochrome B mtDNA) hypothesized the existence of multiple closely related populations of sand fly species (cryptic species) that would exhibit distinct vectorial capacities. This work studies morphotypic diversity using traditional and geometric morphometry analyses carried out on Ph. sergenti's wings from central Morocco, where active L. tropica transmission occurs for 30 years. Methods: Descriptive characteristics (size and shape) of the right wings were measured in Ph. sergenti’s specimens collected from fourteen stations in central Morocco. Both traditional and geometric morphometry methods were used to analyse geographic variations in Ph. sergenti wing’s size and shape. Results: These analyses support the existence of distinct Ph. sergenti populations, enlightening significant phenotypic variations of Ph. sergenti’s wings, regarding their size and shape, depending on geographic origin. In addition, tradi­tional and geometric morphometric analyses of the wing’s length, centroid size, β, ɵ, and γ distances allowed clear dis­crimination of Ph. sergenti sub-populations. Conclusion: These data pinpoint the adaptative ability of Ph. sergenti to local environmental conditions. Additional studies are now required to further shed light on the genetic structure of Ph. sergenti populations in Morocco

Habitat preferences of phlebotomine sand flies (Diptera: Psychodidae) in Southwestern Morocco

Data from a 4-yr study of phlebotomine sand flies in 44 stations in southwestern Morocco are presented. Correspondence analysis is used to describe the habitat preferences of nine sand fly species on a meso-scale (station) and a micro-scale (biotope). This work, based on highly diversified data from various stations, comprising several biotopes sampled during different seasons, provides new information on the ecology of sand flies in southwestern Morocco. It highlights many natural and artificial factors that affect the abundance and distribution of sand fly populations. The impact of climate, urbanization, proximity of humans and domestic animals, organic matter in the soil, shelter, and vegetation type is discussed

Nocturnal activity of phlebotomine sandflies (Diptera : Psychodidae) in a cutaneous leishmaniasis focus in Chichaoua, Morocco

The nocturnal activity of phlebotomine sandflies (Diptera: Psychodidae) was studied "at an epidemic focus" on human cutaneous leishmaniasis due to Leishmania tropica Wright in Chichaoua province, in Morocco. Sandflies were collected using light and sticky-paper traps changed at 2-h intervals, inside and around houses, in August and October 2004. Overall, 633 sandflies, belonging to six species of Phlebotomus and three of Sergentomyia, were collected. Sandfly activity was nocturnal and higher at twilight. Several activity patterns were observed according to the species. Phlebotomus (Paraphlebotomus) sergenti Parrot, 1917, the suspected vector of L. tropica in this focus, was caught during each collection performed from 1900 to 0500 hours, the numbers of species caught peaked at 1900 2100 hours. There were seasonal variations of the nocturnal activity, which could be related to the variations in temperature and relative humidity

Thirty years of cutaneous leishmaniasis in Tadla-Azilal focus, Morocco

Cutaneous leishmaniasis (CL) due to Leishmania tropica is a major health problem in Tadla-Azilal focus, Morocco, where the first case was registered, here, in one locality named Tanant in 1987. So far, CL remains endemic and largely widespread. The objective of this study was to analyze the current eco-epidemiological situation after thirty years of active transmission. Data used are the official ones, obtained from the Moroccan Ministry of Health.Between 1998 and 2015, 5518 CL cases were registered in three provinces; Azilal, Béni Mellal and Fquih Ben Salah. CL has spread, from the historical focus in Azilal along two axes; one to the Northeastern Mountains and the other to the Northwestern plains.CL infected both genders and all ages, with large number of women (53%) and children (75% had <9 years old). More interestingly, age range at risk was larger within females than males, and this difference was more pronounced in Fquih Ben Salah where the disease had newly emerged. Similarly, age ranges at risk were larger and fluctuated significantly each year particularly in new emerging areas in Béni Mellal and Fquih Ben Salah. All these variations may testify, at least in part, to the process of acquiring immunity.Altitude structured CL spatiotemporal distribution. CL was more prevalent in two altitudes ranges; 400–500 m and 800–900 m. The situation and duration of period of diagnostic of CL varied largely according to the altitude. These different scenarios could be related to the seasonal dynamics of vector populations. Keywords: Cutaneous leishmaniasis, Age structure, Space-time distribution, Altitude, Tadla-Azilal, Morocc

Population Genetics of Phlebotomus papatasi from Endemic and Nonendemic Areas for Zoonotic Cutaneous Leishmaniasis in Morocco, as Revealed by Cytochrome Oxidase Gene Subunit I Sequencing

International audienceZoonotic cutaneous leishmaniasis (ZCL) caused by Leishmania major Yakimoff & Shokhor and transmitted by Phlebotomus papatasi (Scopoli) is a public health concern in Morocco. The disease is endemic mainly in pre-Saharan regions on the southern slope of the High Atlas Mountains. The northern slope of the High Atlas Mountains and the arid plains of central Morocco remain nonendemic and are currently considered high risk for ZCL. Here we investigate and compare the population genetic structure of P. papatasi populations sampled in various habitats in historical foci and non-endemic ZCL areas. A fragment of the mtDNA cytochrome oxidase I (CO1) gene was amplified and sequenced in 59 individuals from 10 P. papatasi populations. Haplotype diversity was probed, a median-joining network was generated (FST) and molecular variance (AMOVA) were analyzed. Overall, we identified 28 haplotypes with 32 distinct segregating sites, of which seven are parsimony informative. The rate of private haplotypes was high; 20 haplotypes (71.4%) are private ones and exclusive to a single population. The phylogenetic tree and the network reconstructed highlight a genetic structuration of these populations in two well defined groups: Ouarzazate (or endemic areas) and Non-Ouarzazate (or nonendemic areas). These groups are separated by the High Atlas Mountains. Overall, our study highlights differences in terms of population genetics between ZCL endemic and non-endemic areas. To what extent such differences would impact the transmission of L. major by natural P. papatasi population remains to be investigated

Preliminary study on the galectin molecular diversity in Moroccoan Phlebotomus papatasi sandfly populations

Context: Galactose binding protein (PpGalec) plays an important role in the specificity of Phlebotomus papatasi sandfly for Leishmania major. The molecular diversity of this ligand is currently unknown but might have some influence on the ability of PpGaleC to efficiently recognize L. major in natural sandfly populations. Objective: To explore the molecular diversity of the P papatasi Galectin gene (PpGalec) in natural sandfly population of Morocco. Results & conclusions: Sequence variations of PpGalec was analyzed in 31 P papatasi specimens collected from endemic and non-endemic zoonotic cutaneous leishmaniasis foci of Morocco. Among the 211 amino acid positions analyzed, 11 are subjected to mutation. Interestingly, we observe that one mutation directly affect an amino acid known to be involved in the substrate recognition by galectin. The repercussion of this polymorphism on the capacity of the galectin to efficiently bind the L. major Lipophosphoglycane (LPG) awaits further investigations

Malformations of the genitalia in male Phlebotomus papatasi (Scopoli) (Diptera : Psychodidae)

Phlebotomus papatasi (Scopoli, 1786) (Diptera: Psychodidae) is a major vector of Leishmania major (Kinetoplastida: Trypanosomatidae), a causative agent of zoonotic cutaneous leishmaniasis. Morphological characters of sand fly genitalia are key indicators for species identification. Various anomalies affecting male genitalia have been previously described. We take advantage of a large sand flies survey conducted in 32 stations in Central and Southern Morocco to systematically quantify the prevalence and spatial distribution of malformations affecting the genitalia of P. papatasi. Among 597 examined males, 122 were abnormal (20.4%). Malformations were widespread and largely concerned the number of spines in the lateral lobes and in the styles. Asymmetrical anomalies in lateral lobes were common. Correspondence analysis of our results highlighted the symmetrical anomalies observed in the lateral lobes, and abnormal styles of the male genitalia were found to be associated with environmental disturbances since they were prevalent in sewage dumps. Journal of Vector Ecology 35 (1): 13-19. 2010

Bacterial flora as indicated by PCR-temperature gradient gel electrophoresis (TGGE) of 16S rDNA gene fragments from isolated guts of phlebotomine sand flies (Diptera : Psychodidae)

In this study, we tested the capacity of Temperature Gradient Gel Electrophoresis (TGGE)-based fingerprinting of 16S rDNA PCR fragments to assess bacterial composition in a single isolated sand fly gut. Bacterial content was studied in different life stages of a laboratory-reared colony of Phlebotomus duboscqi and in a wild-caught Phlebotomus papatasi population. Our study demonstrates that a major reorganization in the gut bacterial community occurs during metamorphosis of sand flies. Chloroflexi spp. was dominant in the guts of pre-imaginal stages, although Microbacterium spp. and another as yet unidentified bacteria were detected in the gut of the adult specimen. Interestingly, Microbacterium spp. was also found in all the adult guts of both species. We demonstrate that the analysis of bacterial diversity in an individualized sand fly gut is possible with fingerprinting of 16S rDNA. The use of such methodology, in conjunction with other culture-based methods, will be of great help in investigating the behavior of the Leishmania-bacterial community in an ecological context

Genetic structure of Phlebotomus (Larroussius) ariasi populations, the vector of Leishmania infantum in the western Mediterranean : epidemiological implications

In recent years there has been growing interest in analyzing the geographical variations between populations of different Phlebotomus spp. by comparing the sequences of various genes However, little is known about the genetic structure of Phlebotonnis arias,. In this study, we were able to sequence a fragment of the mitochondrial Cyt b gene in 133 sandflies morphologically identified as P anasi and proceeding from a wide geographical range covering 35 locations in 11 different regions from five countries. The intra-specific diversity of P. anasi is high, with 45 haplotypes differing from each other by one to 26 bases and they are distributed in two mitochondria' lineages, one limited geographically to Algeria and the other widely dispersed across Mediterranean countries The Algerian lineage is characterized by having 13 fixed polymorphisms and is made up of one sole haplotype. The European/Moroccan P. anasz lineage is characterized by being made up of a great diversity of haplotypes (44) which display some geographical structuring. This could be one of the multiple factors involved in the epidemiological heterogeneity of the foci of leishmaniasis Phlebotomus choral, is the sister group of European/Moroccan P. anasi. The separation of the Algerian haplotype, H45, from the rest of the specimens, European/Moroccan P anon and P chadln, is well supported by the bootstrap analysis (C) 2010 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved