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This paper is the first large-scale molecular phylogenetic study on filarial parasites (family Onchocercidae) which includes 16 spe­ cies of 6 genera : Brugia beaveri Ash et Little, 1964 ; B. buckleyiDissanaike et Paramananthan, 1961 ; B. malayi (Brug, 1927) Buckley, 1960; B. pahangi (Buckley et Edeson, 1956) Buckley, 1960; B. patei (Buckley, Nelson et Heisch, 1958) Buckley, 1960; B. timori Partono etal, 1977; Wuchereria bancrofti(Cobbold, 1877) Seurat, 1921; W. kalimantani Palmieri , Purnomo, Dennis and Marwoto, 1980; Mansonella perstans(Manson, 1891 ) Eberhard et Orihel, 1 9 8 4 ; Loa loa, Stiles, 1905; Onchocerca volvulus (Leuckart, 1983) Railliet et Henry, 1910; O. ochengi Bwangamoi, 1969; O. gutturosa Neumann, 1910; Dirofilaria immitis (Leidy, 1856) Railliet et Henry, 1911 ; Acanthocheilonema viteae (Krepkogorskaya, 1933) Bain, Baker et Chabaud, 1982 and Litomosoides sigmodontis Chandler, 1931. 5S rRNA gene spacer region sequence data were collec­ ted by PCR, cloning and dideoxy sequencing. The 5S rRNA gene spacer region sequences were aligned and analyzed by maxi­ mum parsimony algorithms, distance methods and maximum likeli­ hood methods to construct phylogenetic trees. Bootstrap analysis was used to test the robustness of the different phylogenetic reconstructions. The data indicated that 5S spacer region sequences are highly conserved within species yet differ signifi­ cantly between species. Spliced leader sequences were observed in all of the 5S rDNA spacers with no sequence variation, although flanking region sequence and length heterogeneity was observed even within species. All of the various tree-building methods gave very similar results. This study identified four clades which are strongly supported by bootstrap analysis: the Brugiaclade; the Wuchereria clade; the Brugia-Wuchereria clade and the Onchocerca clade. The analyses indicated that L. sigmodontisand A. viteae may be the most primitive among the 16 species studied. The data did not show any close relationship betweenLoa loa and D. immitis presently classified in the same subfamily, and the constitution of the Dirofilariinae subfamily is questionable.


phylogenetics, ribosomal genes, 5S rDNA, spacers, PCR, molecular cloning, spliced leader sequence, Brugia malayi, B. pahangi, B. timori, B. patei, B. beaveri, B. buckleyi, Wuchereria bancrofti, W. kaliman­ tani, Mansonella perstans, Loa loa, Onchocerca volvulus, O. ochengi, O. gutturosa, Dirofilaria immitis, Acanthocheilonema viteae, Litomosoides sigmo­ dontis



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