ABSTRACT
The idea of the existence of Phlebotomus (Larroussius) neglectus (Diptera: Psychodidae) Tonnoir, 1921 in Iran and the skepticism about the existence of Phlebotomus major s.str. Annandale, 1910 had been grown recently in the country. This study reports a combined analysis of mitochondrial and ribosomal DNA target regions of P. major s.l.Annandale, 1910, specimens collected from different parts of Iran. Two different morphotypes were found among the collected samples based on the shape of the aedeagus, ventrally located hairs of the coxite, and parameral sheets. One morphotype seemed similar to P. neglectus Tonnoir 1921 or P. major krimensis Perfiliv1966 (called here MI.N.K.); the other one was similar to P. neglectus and to some extent to P. notus Artemiev & Neronov 1984 (here called MII.N.NO). Cytochrome B, elongation factor 1-alpha, and internal transcribed spacer II loci were amplified, sequenced, and characterized. High sequence homology (98-100%) was observed between P. neglectus and these morphotypes, and phylogenetic analysis was also concordant. Phlebotomus neglectus sequences available in GenBank are located as the sister group of sequences here, particularly near to morphotype MII.N.NO. Moreover, ITS2 locus provides the maximum resolution for differentiation of two morphotypes. Based on achieving results, although a strong support for the presence of P. neglectus was provided, but it is too early to say that P. major s.str. does/does not exist in Iran. This question could be resolved by studying more samples and, most importantly, by comparing the topotypes of P. neglectus and P. major s. str. if possible in the future.
Subject(s)
Phlebotomus , Animals , Cytochromes b/genetics , DNA, Ribosomal/genetics , Genes, Insect , Genetic Loci , Genetic Markers , Insect Vectors/anatomy & histology , Insect Vectors/genetics , Iran , Phlebotomus/anatomy & histology , Phlebotomus/genetics , Phylogeny , Psychodidae/anatomy & histology , Psychodidae/genetics , Species SpecificityABSTRACT
A family of reticulocyte-binding proteins of Plasmodium vivax (PvRBP) is localised at the apical pole of the merozoites and appears to bind to reticulocytes specifically and has also been involved in identifying host cells. Protein component produced by the Pvrbp2c gene is highly antigenic. The aim of this study was to detect the genetic diversity in the Pvrbp2c gene of Iranian P. vivax field isolates using the polymerase chain reaction- restricted fragment length polymorphism (PCR-RFLP) technique. A total of 79 P. vivax malaria patients with fever participated in the study. Alu1 and Apo1 restriction enzymes were independently used to identify allelic variants of the Pvrbp2c gene. All of the samples exhibited a single band of about 2 Kb in nested PCR. Among 79 P. vivax field isolates in the RFLP with Apo1 and Alu1 restriction enzymes, 15 and nine patterns were observed, respectively. In total, 24 various patterns were detected from the combined findings of both Alu1 and Apo1 fragments in RFLP. This study revealed that Pvrbp2c has genetic diversity in southeast Iran. Genotyping of Pvrbp2c not only shows the heterogeneity of P. vivax but also provides important information that could be used to control vivax malaria.
ABSTRACT
A family of reticulocyte-binding proteins of Plasmodium vivax (PvRBP) is localised at the apical pole of the merozoites and appears to bind to reticulocytes specifically and has also been involved in identifying host cells. Protein component produced by the Pvrbp2c gene is highly antigenic. The aim of this study was to detect the genetic diversity in the Pvrbp2c gene of Iranian P. vivax field isolates using the polymerase chain reaction- restricted fragment length polymorphism (PCR-RFLP) technique. A total of 79 P. vivax malaria patients with fever participated in the study. Alu1 and Apo1 restriction enzymes were independently used to identify allelic variants of the Pvrbp2c gene. All of the samples exhibited a single band of about 2 Kb in nested PCR. Among 79 P. vivax field isolates in the RFLP with Apo1 and Alu1 restriction enzymes, 15 and nine patterns were observed, respectively. In total, 24 various patterns were detected from the combined findings of both Alu1 and Apo1 fragments in RFLP. This study revealed that Pvrbp2c has genetic diversity in southeast Iran. Genotyping of Pvrbp2c not only shows the heterogeneity of P. vivax but also provides important information that could be used to control vivax malaria.
ABSTRACT
A comparative morphological and molecular study was carried out on 11 different populations of Phlebotomus (Paraphlebotomus) caucasicus Marzinovsky 1917 caught in 7 provinces in Iran (2004-2005). Differences in the implantation level of the two distal spines of the style, the number of setae of the basal lobe of coxite, and the length of the third antennal segment, revealed the existence of two morphotypes within P. (Pa) caucasicus, a species having a confused history if we take into account an unclear synonymisation with Phlebotomus (Paraphlebotomus) grimmi Porchinsky 1874. Sequencing of mtDNA (a fragment of cytchromeB gene, tRNA for serine gene and a fragment of NADH1 gene) and Neighbour-Joining analysis showed a partial correlation between morphotypes and haplotypes. We also found a correlation between the latter and the geographical origin of the specimens. These results need further studies in order to appreciate the role of each morphotype/haplotype in the transmission of Leishmania major.
