University of Khartoum

Molecular and Taxonomic Studies on Phlebotomine Sandflies of Medical Importance in Sudan

Molecular and Taxonomic Studies on Phlebotomine Sandflies of Medical Importance in Sudan

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Title: Molecular and Taxonomic Studies on Phlebotomine Sandflies of Medical Importance in Sudan
Author: Saeed, Sara Abdalla Khidir
Abstract: This study aimed to develop a unique molecular tool combining single step of multiplex PCR and restriction fragments length polymorphism (RFLP) that targeting Internal transcribed spacer 2 (ITS2) of the rDNA to identify Phlebotomus species of medically importance collected from different geographical sites from Sudan. The intraspecific morphological differences were studied among the populations of Phlebotomus papatasi and Phlebotomus orientalis using land mark- geometric morphometric analysis on 13 landmarks and the width of the right wing. Fields 'trips were conducted during 2012 and 2015 in five sites of Sudan for the collection of Sandfly samples. The specimens were collected using CDC miniature light traps, sticky paper traps (Gastor oil), and animal baited traps. The total number of sandfly specimens collected was 1200 and about 263 specimens were found belonging to five Phlebotomus species while the rest were Sergentomyia species. Among the populations studied of P. orientalis, landmark 13 showed significant differences between the males of four populations (One-Way ANOVA; P value=0.0175, DF=3) and also the females of four populations (One-Way ANOVA; P value =0.00, DF=3). In P. papatasi, landmark 13 showed significant differences among the males of seven populations (One-Way ANOVA; P value =0.0052, DF=6), also the wing width showed significant differences among the populations studied (One-Way ANOVA; P value =0.0078, DF=6). The ITS2 region showed the same size of banding pattern of 490 bp in all Phlebotomus species studied and 590 bp in Sergentomyia species. High ATs contents were reported. The Phlebotomus species showed high identity of their ITS2 sequences between them that ranging from 91.6% to 52.8%, while Sergentomyia species showed less identity percentage of 34%. The VIII multiplex PCR successfully differentiate between P. papatasi and P. orientalis. The application of RFLP using the two restriction enzymes Alu1and DraI showed more advantages than multiplex PCR in the differentiation between the closely related species P. papatasi, P. bergeroti and P. duboscqi. The combination of PCR-RFLP based method considered to be as powerful tool in sandfly taxonomy and research. Both morphological and molecular data obtained from this study showed the high similarity among the populations of each of the two vector species P. papatasi and P. orientalis. P. duboscqi due to the high similarity of their ITS2 sequences. The third restriction enzyme Ms1I failed to distinguish between P. papatasi and P. bergeroti. The combination of PCR-RFLP based method considered to be as powerful tool in sandfly taxonomy and research. Both morphological and molecular data obtained from this study showed the high similarity a mong the populations of each of the two vector species P. papatasi and P. orientalis. The few differences reported in both morphology or molecular data might be attributed to the differences of the environmental factors within the microhabitats for each population.
URI: http://khartoumspace.uofk.edu/123456789/26491


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