Molecular Characterization of Gregarines from Sand Flies (Diptera: Psychodidae) and Description of Psychodiella n. g. (Apicomplexa: Gregarinida)

Abstract

Sand fly and mosquito gregarines have been lumped for a long time in the single genus Ascogregarina and on the basis of their morphological characters and the lack of merogony been placed into the eugregarine family Lecudinidae. Phylogenetic analyses performed in this study clearly demonstrated paraphyly of the current genus Ascogregarina and revealed disparate phylogenetic positions of gregarines parasitizing mosquitoes and gregarines retrieved from sand flies. Therefore, we reclassified the genus Ascogregarina and created a new genus Psychodiella to accommodate gregarines from sand flies. The genus Psychodiella is distinguished from all other related gregarine genera by the characteristic localization of oocysts in accessory glands of female hosts, distinctive nucleotide sequences of the small subunit rDNA, and host specificity to flies belonging to the subfamily Phlebotominae. The genus comprises three described species: the type species for the new genus--Psychodiella chagasi (Adler and Mayrink 1961) n. comb., Psychodiella mackiei (Shortt and Swaminath 1927) n. comb., and Psychodiella saraviae (Ostrovska, Warburg, and Montoya-Lerma 1990) n. comb. Its creation is additionally supported by sequencing data from other gregarine species originating from the sand fly Phlebotomus sergenti. In the evolutionary context, both genera of gregarines from mosquitoes (Ascogregarina) and sand flies (Psychodiella) have a close relationship to neogregarines; the genera represent clades distinct from the other previously sequenced gregarines.

The life cycle and host specificity of Psychodiella sergenti n. sp. and Ps. tobbi n. sp. (Protozoa: Apicomplexa) in sand flies Phlebotomus sergenti and Ph. tobbi (Diptera: Psychodidae)

“…from Ph. sergenti) originated from our previous study (Votý pka et al, 2009) and are deposited in GenBank under accessional numbers FJ865354 and FJ865355, respectively. Data set containing all gregarine sequences of small subunit ribosomal DNAs accessible at the time of the study was used to establish the phylogenetic position of both newly described species of sand fly gregarines.…”

“…Natural sand fly host Experimentally infected sand fly host amplification, and sequencing of SSU rDNA were described by Votý pka et al (2009). The primers used for sequencing were: the vector primers M13 (F) 5 0 -GTAAAACCACGGCCAG-3 0 and M13 (R) 5 0 -CAGGAAACAGCTATGAC-3 0 and internal primers oriented in both directions 5 0 -AAGACGATCAGATACCG-3 0 , 5 0 -TCGATTCCGGAGAGG-GA-3 0 , 5 0 -CGTCAATTCCTTTAAG-3 0 , 5 0 -GCTGGCACCAGACTTGC-3 0 .…”

“…The systematic status of gregarines has not been resolved and would greatly benefit from the addition of DNA sequences as molecular characters (Leander, 2008). Molecular phylogenetic data demonstrate that gregarines are more divergent than previously assumed and new lineages have emerged from older taxa (Leander, 2008;Votý pka et al, 2009).…”

“…Nematoceran Diptera are usually considered as rare hosts of gregarines and only 12 named species of two relatively morphologically uniform genera inhabit mosquitoes (Ascogregarina) and sand flies (Psychodiella) (Warburg and Ostrovska, 1991;Chen, 1999;Perkins et al, 2000;Votý pka et al, 2009). The genus Psychodiella encompasses three named species of the aseptate gregarines (order Eugregarinorida according to Perkins et al (2000)) that parasitize sand flies.…”

Identificación molecular de la mosquilla del brote Prodiplosis sp. Gagné en los cultivos de Asparagus officinalis L. por amplificación parcial del gen citocromo oxidasa I

“…Los marcadores moleculares más utilizados para la identificación de organismos son el ADN genómico (ADN ribosomal, ADNr), también denominado marcador nuclear, así como los genes mitocondriales (ADN mitocondrial, ADNmt), ambos marcadores emplean la PCR (Polymerase Chain Reaction, por sus siglas en inglés) acoplada a la secuenciación para la identificación de regiones que permitan la identificación lo más exacta posible del organismo en estudio. Votypka et al, 2009, mediante análisis filogenéticos utilizó la subunidad pequeña del ADNr para la diferenciación de las gregarinas asociadas a mosquitos de las moscas de la arena. Ruiz et al, 2010, mediante Prodiplosis sp.…”