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

Abstract

BackgroundNew foci of human CL caused by strains of the Leishmania donovani (L. donovani) complex have been recently described in Cyprus and the Çukurova region in Turkey (L. infantum) situated 150 km north of Cyprus. Cypriot strains were typed by Multilocus Enzyme Electrophoresis (MLEE) using the Montpellier (MON) system as L. donovani zymodeme MON-37. However, multilocus microsatellite typing (MLMT) has shown that this zymodeme is paraphyletic; composed of distantly related genetic subgroups of different geographical origin. Consequently the origin of the Cypriot strains remained enigmatic.Methodology/Principal FindingsThe Cypriot strains were compared with a set of Turkish isolates obtained from a CL patient and sand fly vectors in south-east Turkey (Çukurova region; CUK strains) and from a VL patient in the south-west (Kuşadasi; EP59 strain). These Turkish strains were initially analyzed using the K26-PCR assay that discriminates MON-1 strains by their amplicon size. In line with previous DNA-based data, the strains were inferred to the L. donovani complex and characterized as non MON-1. For these strains MLEE typing revealed two novel zymodemes; L. donovani MON-309 (CUK strains) and MON-308 (EP59). A population genetic analysis of the Turkish isolates was performed using 14 hyper-variable microsatellite loci. The genotypic profiles of 68 previously analyzed L. donovani complex strains from major endemic regions were included for comparison. Population structures were inferred by combination of Bayesian model-based and distance-based approaches. MLMT placed the Turkish and Cypriot strains in a subclade of a newly discovered, genetically distinct L. infantum monophyletic group, suggesting that the Cypriot strains may originate from Turkey.ConclusionThe discovery of a genetically distinct L. infantum monophyletic group in the south-eastern Mediterranean stresses the importance of species genetic characterization towards better understanding, monitoring and controlling the spread of leishmaniasis in this region.

Is Predominant Clonal Evolution a Common Evolutionary Adaptation to Parasitism in Pathogenic Parasitic Protozoa, Fungi, Bacteria, and Viruses?

“…Near-clades are also observed in the L. infantum complex. In Turkey and Cyprus, they show concordance between (1) STRUCTURE, NJ trees and factorial correspondence analysis and (2) between K26-PCR, MLEE and microsatellites (Gouzelou et al, 2012). Near-clading agreement between Internal Transcribed Spacer, MLEE, MLST, miniexon typing and gp63 intergenic RFLP has also been observed (Mauricio et al, 2006).…”

“…LD between different genetic markers in the L. infantum complex is clear (Gouzelou et al, 2012;Kuhls et al, 2008;Mauricio et al, 2006). Some microsatellite MLGs of this species are sampled in both the Ancient and the New World .…”

Leishmaniasis in Sri Lanka: a decade old story

“…are also being continuously reported. 5,6 Sri Lanka is the newest reported focus of leishmaniasis in the Indian subcontinent. 7 The disease is caused by the most virulent visceralizing species, L.donovani.…”

Response to Rougeron et al.: Leishmania population genetics: clonality, selfing and aneuploidy

“…The limited discrepancy between the two markers is easily explained by MLEE homoplasy, although it may also indicate occasional recombination, which is compatible with the PCE model [2,3]. Most times, MLEE and DNA markers exhibit a fair association: all Leishmania infantum strains MON 1 correspond to a unique microsatellite cluster, which comprises several genotypes ( [6] for example). This is not a discrepancy; it shows only that microsatellites have a stronger resolution power than MLEE.…”