Characterization of simple sequence repeats (SSRs) from Phlebotomus papatasi (Diptera: Psychodidae) expressed sequence tags (ESTs)

<p>Abstract</p> <p>Background</p> <p><it>Phlebotomus papatasi </it>is a natural vector of <it>Leishmania major</it>, which causes cutaneous leishmaniasis in many countries. Simple sequence repeats (SSRs), or microsatellites, are common in eukaryotic genomes and are short, repeated nucleotide sequence elements arrayed in tandem and flanked by non-repetitive regions. The enrichment methods used previously for finding new microsatellite loci in sand flies remain laborious and time consuming; <it>in silico </it>mining, which includes retrieval and screening of microsatellites from large amounts of sequence data from sequence data bases using microsatellite search tools can yield many new candidate markers.</p> <p>Results</p> <p>Simple sequence repeats (SSRs) were characterized in <it>P. papatasi </it>expressed sequence tags (ESTs) derived from a public database, National Center for Biotechnology Information (NCBI). A total of 42,784 sequences were mined, and 1,499 SSRs were identified with a frequency of 3.5% and an average density of 15.55 kb per SSR. Dinucleotide motifs were the most common SSRs, accounting for 67% followed by tri-, tetra-, and penta-nucleotide repeats, accounting for 31.1%, 1.5%, and 0.1%, respectively. The length of microsatellites varied from 5 to 16 repeats. Dinucleotide types; AG and CT have the highest frequency. Dinucleotide SSR-ESTs are relatively biased toward an excess of (AX)n repeats and a low GC base content. Forty primer pairs were designed based on motif lengths for further experimental validation.</p> <p>Conclusion</p> <p>The first large-scale survey of SSRs derived from <it>P. papatasi </it>is presented; dinucleotide SSRs identified are more frequent than other types. EST data mining is an effective strategy to identify functional microsatellites in <it>P. papatasi</it>.</p

Multilocus Microsatellite Typing (MLMT) of Strains from Turkey and Cyprus Reveals a Novel Monophyletic L. donovani Sensu Lato Group

In eastern Mediterranean, leishmaniasis represents a major public health problem with considerable impact on morbidity and potential to spread. Cutaneous leishmaniasis (CL) caused by L. major or L. tropica accounts for most cases in this region although visceral leishmaniasis (VL) caused by L. infantum is also common. New foci of human CL caused by L. donovani complex strains were recently described in Cyprus and Turkey. Herein we analyzed Turkish strains from human CL foci in Çukurova region (north of Cyprus) and a human VL case in Kuşadasi. These were compared to Cypriot strains that were previously typed by Multilocus Enzyme Electrophoresis (MLEE) as L. donovani MON-37. Nevertheless, they were found genetically distinct from MON-37 strains of other regions and therefore their origin remained enigmatic. A population study was performed by Multilocus Microsatellite Typing (MLMT) and the profile of the Turkish strains was compared to previously analyzed L. donovani complex strains. Our results revealed close genetic relationship between Turkish and Cypriot strains, which form a genetically distinct L. infantum monophyletic group, suggesting that Cypriot strains may originate from Turkey. Our analysis indicates that the epidemiology of leishmaniasis in this region is more complicated than originally thought

The Genetic Structure of Leishmania infantum Populations in Brazil and Its Possible Association with the Transmission Cycle of Visceral Leishmaniasis

Leishmania infantum is the etiologic agent of visceral leishmaniasis (VL) in the Americas, Mediterranean basin and West and Central Asia. Although the geographic structure of L. infantum populations from the Old World have been described, few studies have addressed the population structure of this parasite in the Neotropical region. We employed 14 microsatellites to analyze the population structure of the L. infantum strains isolated from humans and dogs from most of the Brazilian states endemic for VL and from Paraguay. The results indicate a low genetic diversity, high inbreeding estimates and a depletion of heterozygotes, which together indicate a predominantly clonal breeding system, but signs of sexual events are also present. Three populations were identified from the clustering analysis, and they were well supported by F statistics inferences and partially corroborated by distance-based. POP1 (111 strains) was observed in all but one endemic area. POP2 (31 strains) is also well-dispersed, but it was the predominant population in Mato Grosso (MT). POP3 (31 strains) was less dispersed, and it was observed primarily in Mato Grosso do Sul (MS). Strains originated from an outbreak of canine VL in Southern Brazil were grouped in POP1 with those from Paraguay, which corroborates the hypothesis of dispersal from Northeastern Argentina and Paraguay. The distribution of VL in MS seems to follow the west-east construction of the Bolivia-Brazil pipeline from Corumbá municipality. This may have resulted in a strong association of POP3 and Lutzomyia cruzi, which is the main VL vector in Corumbá, and a dispersion of this population in this region that was shaped by human interference. This vector also occurs in MT and may influence the structure of POP2. This paper presents significant advances in the understanding of the population structure of L. infantum in Brazil and its association with eco-epidemiological aspects of VL

Comparative Microsatellite Typing of New World Leishmania infantum Reveals Low Heterogeneity among Populations and Its Recent Old World Origin

Leishmania infantum (syn. L. chagasi) is the causative agent of visceral leishmaniasis (VL) in the New World (NW) with endemic regions extending from southern USA to northern Argentina. The two hypotheses about the origin of VL in the NW suggest (1) recent importation of L. infantum from the Old World (OW), or (2) an indigenous origin and a distinct taxonomic rank for the NW parasite. Multilocus microsatellite typing was applied in a survey of 98 L. infantum isolates from different NW foci. The microsatellite profiles obtained were compared to those of 308 L. infantum and 20 L. donovani strains from OW countries previously assigned to well-defined populations. Two main populations were identified for both NW and OW L. infantum. Most of the NW strains belonged to population 1, which corresponded to the OW MON-1 population. However, the NW population was much more homogeneous. A second, more heterogeneous, population comprised most Caribbean strains and corresponded to the OW non-MON-1 population. All Brazilian L. infantum strains belonged to population 1, although they represented 61% of the sample and originated from 9 states. Population analysis including the OW L. infantum populations indicated that the NW strains were more similar to MON-1 and non-MON-1 sub-populations of L. infantum from southwest Europe, than to any other OW sub-population. Moreover, similarity between NW and Southwest European L. infantum was higher than between OW L. infantum from distinct parts of the Mediterranean region, Middle East and Central Asia. No correlation was found between NW L. infantum genotypes and clinical picture or host background. This study represents the first continent-wide analysis of NW L. infantum population structure. It confirmed that the agent of VL in the NW is L. infantum and that the parasite has been recently imported multiple times to the NW from southwest Europe

Comportement des poteaux composites en profils creux en acier remplis de béton = Behavior of composite columns in hollow steel section filled with concrete

Le but de cet article, est la determination des rigidites flexionnelles EIx et EIy d’fune section mixte acier beton et plus precisement d’fun poteau en tube d’facier de section rectangulaire, remplie de beton, sollicitee a la flexion bi-axiale (N, Mx et My). L’festimation des rigidites sera faite a partir d’fune approche theorique par une analyse du poteau en elements finis (element barre a 4 degres de liberte), basee sur les conditions d’fequilibres a mi-portee en utilisant la relation moment-courbure (M–Φ) de l’felement deforme par application de l’fequation suivante: EI=M/Φ. Le comportement des materiaux est celui comme adopte par les reglements Eurocode 2 et 3, respectivement pour le beton et l’facier. Afin de valider l’fapproche theorique utilisee dans cette etude, deux comparaisons ont ete faites : une premiere permettant de comparer les resultats des rigidites determinees par les relations moments courbures et celles calculees par l’fEurocode 4 et une deuxieme comparaison entre les charges de ruines de deux poteaux de grandeurs natures avec ceux testes au laboratoire [2]. Au vu des resultats obtenus, nous pouvons conclure que l’approche théorique utilisée dans cette étude ainsi que les modèles de comportement des matériaux sont adéquats pour ce genre de problème

Comportement des poteaux composites en profils creux en acier remplis de béton

published by EDP Sciences.The purpose of this paper is the determination of flexural stiffness EIx and EIy of a concrete filled rectangular cross section of a composite steel column, under biaxial bending (N, Mx and M y). The rigidities will be estimated from a theoretical approach using a finite element analysis (element bar with 4 degrees of freedom), based on the equilibrium conditions at mid-span using the moment-curvature relationships (M-Φ) of the deformed element by applying the following equation: EI=M/Φ. The material behavior is the one adopted by Eurocode 2 and 3, respectively, for concrete and steel. To validate the theoretical approach used, two comparisons were made: one comparing the rigidities given by the moment-curvature relations versus those calculated by the Eurocode 4, and a second comparison between failure loads of two columns scale one with those tested in the laboratory [2]. Based on the results obtained, we may conclude that the theoretical approach used in this study and the models of behavior of materials are suitable for such problem

Comportement des poteaux composites en profils creux en acier remplis de béton

Le but de cet article, est la determination des rigidites flexionnelles EIx et EIy d’fune section mixte acier beton et plus precisement d’fun poteau en tube d’facier de section rectangulaire, remplie de beton, sollicitee a la flexion bi-axiale (N, Mx et My). L’festimation des rigidites sera faite a partir d’fune approche theorique par une analyse du poteau en elements finis (element barre a 4 degres de liberte), basee sur les conditions d’fequilibres a mi-portee en utilisant la relation moment-courbure (M–Φ) de l’felement deforme par application de l’fequation suivante: EI=M/Φ. Le comportement des materiaux est celui comme adopte par les reglements Eurocode 2 et 3, respectivement pour le beton et l’facier. Afin de valider l’fapproche theorique utilisee dans cette etude, deux comparaisons ont ete faites : une premiere permettant de comparer les resultats des rigidites determinees par les relations moments courbures et celles calculees par l’fEurocode 4 et une deuxieme comparaison entre les charges de ruines de deux poteaux de grandeurs natures avec ceux testes au laboratoire [2]. Au vu des resultats obtenus, nous pouvons conclure que l’approche théorique utilisée dans cette étude ainsi que les modèles de comportement des matériaux sont adéquats pour ce genre de problèmes

Composition chimique et activité antibactérienne des Huiles Essentielles des aiguilles de Pinus halepensis Mill. du Nord est Algérien

L'huile essentielle d’aiguilles de Pinus halepensis Mill., récoltées dans la forêt du lac Mellah situé dans le Parc National d’El Kala et la forêt Zaarouria (Souk Ahras), obtenue par hydrodistillation donne respectivement un rendement de 0,81% et de 0,3%. L’huile essentielle des aiguilles du lac Mellah comprend 16 constituants. Les majoritaires sont, le β-caryophyllène (31,89%), l’α-pinène (24,41%) et le β-myrcène (19,38%). Celle de Zaarouria, comprend 16 composés représentés essentiellement par l’α-pinène (21,01%), le β-caryophyllène (31,14%) et l’α-terpinolène (9,43%). Ces huiles essentielles testées in vitro sur trois souches antibio-résistantes (Staphylococcus aureus, Pseudomonas aeroginosa, et Escherichia coli), souvent responsables d’infection nosocomiale ont montré un pouvoir antibactérien.Mots clés : huile essentielle - Pinus halepensis Mill. - activité antibactérienne - CG/SM