Limits...
First isolation of Leishmania from Northern Thailand: case report, identification as Leishmania martiniquensis and phylogenetic position within the Leishmania enriettii complex.

Pothirat T, Tantiworawit A, Chaiwarith R, Jariyapan N, Wannasan A, Siriyasatien P, Supparatpinyo K, Bates MD, Kwakye-Nuako G, Bates PA - PLoS Negl Trop Dis (2014)

Bottom Line: Here we report a case in a 52-year-old Thai male from northern Thailand, who presented with subacute fever, huge splenomegaly and pancytopenia.This is the first report of visceral leishmaniasis caused by L. martiniquensis from the region.Moreover, the majority of parasites previously identified as "L. siamensis" also appear to be L. martiniquensis.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand.

ABSTRACT
Since 1996, there have been several case reports of autochthonous visceral leishmaniasis in Thailand. Here we report a case in a 52-year-old Thai male from northern Thailand, who presented with subacute fever, huge splenomegaly and pancytopenia. Bone marrow aspiration revealed numerous amastigotes within macrophages. Isolation of Leishmania LSCM1 into culture and DNA sequence analysis (ribosomal RNA ITS-1 and large subunit of RNA polymerase II) revealed the parasites to be members of the Leishmania enriettii complex, and apparently identical to L. martiniquensis previously reported from the Caribbean island of Martinique. This is the first report of visceral leishmaniasis caused by L. martiniquensis from the region. Moreover, the majority of parasites previously identified as "L. siamensis" also appear to be L. martiniquensis.

Show MeSH

Related in: MedlinePlus

Phylogenetic analysis by ML method of Leishmania strain CM1 using RNA Pol II sequences.Tree with 25 species of Leishmania and Endotrypanum monterogeii using Trypanosoma brucei as an outgroup, based on alignment of 1191-1206 homologous nucleotide sequences. AM-2004 is Leishmania from Australia, PCM2 is the “L. siamensis” PCM2 Trang isolate. Bootstrap values from 1000 replicates are given at the nodes.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4256172&req=5

pntd-0003339-g005: Phylogenetic analysis by ML method of Leishmania strain CM1 using RNA Pol II sequences.Tree with 25 species of Leishmania and Endotrypanum monterogeii using Trypanosoma brucei as an outgroup, based on alignment of 1191-1206 homologous nucleotide sequences. AM-2004 is Leishmania from Australia, PCM2 is the “L. siamensis” PCM2 Trang isolate. Bootstrap values from 1000 replicates are given at the nodes.

Mentions: These observations required further investigation, since the ITS-1 sequence is known to be polymorphic [20] and does not contain enough informative sites to be reliable for phylogenetic analysis of Leishmania species. Therefore, a different target was used for this analysis, the large subunit RNA Polymerase II (Pol II) gene (L. major chromosome 31) that has been previously used for Leishmania phylogenetic analyses [14], [16], [21], [22]. LSCM1 DNA was extracted from culture and amplified with RPOF1/RPOR1 and various newly designed primer pairs (see Methods) to generate sequences of the RNA Pol II gene. Additional sequences were also generated for L. martiniquensis, “L. siamensis” (PCM2 Trang strain, [9]) and for L. colombiensis. Analysis was performed on the 25 available Leishmania RNA Pol II sequences (21 existing +4 new sequences from this study), Endotrypanum monterogeii and using Trypanosoma brucei as an outgroup. The resulting Maximum Likelihood (ML) tree is shown in Fig. 5. A topologically identical Neighbour Joining (NJ) tree was also generated (Figure S2). Bootstrapping provided strong support for almost all of the nodes, the only two exceptions were 54% bootstrap support for the clade including subgenera Leishmania/Sauroleishmania/Viannia in the ML tree (Fig. 5), however, this node received 94% support in the NJ tree; and 66% support for the clade including all species except paraleishmania II in the NJ tree (Figure S2), however, this node received 85% support in the ML tree. We also tested alternate outgroups, such as Crithidia fasciculata, Leptomonas costaricensis and L. seymouri, and the topologies of the resulting ML trees were similar to Fig. 5. For example, with C. fasciculata as an outgroup the main difference was that the paraleishmania appear as a monophyletic clade, and consequently the rooting of tree is slightly different (Figure S3).


First isolation of Leishmania from Northern Thailand: case report, identification as Leishmania martiniquensis and phylogenetic position within the Leishmania enriettii complex.

Pothirat T, Tantiworawit A, Chaiwarith R, Jariyapan N, Wannasan A, Siriyasatien P, Supparatpinyo K, Bates MD, Kwakye-Nuako G, Bates PA - PLoS Negl Trop Dis (2014)

Phylogenetic analysis by ML method of Leishmania strain CM1 using RNA Pol II sequences.Tree with 25 species of Leishmania and Endotrypanum monterogeii using Trypanosoma brucei as an outgroup, based on alignment of 1191-1206 homologous nucleotide sequences. AM-2004 is Leishmania from Australia, PCM2 is the “L. siamensis” PCM2 Trang isolate. Bootstrap values from 1000 replicates are given at the nodes.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4256172&req=5

pntd-0003339-g005: Phylogenetic analysis by ML method of Leishmania strain CM1 using RNA Pol II sequences.Tree with 25 species of Leishmania and Endotrypanum monterogeii using Trypanosoma brucei as an outgroup, based on alignment of 1191-1206 homologous nucleotide sequences. AM-2004 is Leishmania from Australia, PCM2 is the “L. siamensis” PCM2 Trang isolate. Bootstrap values from 1000 replicates are given at the nodes.
Mentions: These observations required further investigation, since the ITS-1 sequence is known to be polymorphic [20] and does not contain enough informative sites to be reliable for phylogenetic analysis of Leishmania species. Therefore, a different target was used for this analysis, the large subunit RNA Polymerase II (Pol II) gene (L. major chromosome 31) that has been previously used for Leishmania phylogenetic analyses [14], [16], [21], [22]. LSCM1 DNA was extracted from culture and amplified with RPOF1/RPOR1 and various newly designed primer pairs (see Methods) to generate sequences of the RNA Pol II gene. Additional sequences were also generated for L. martiniquensis, “L. siamensis” (PCM2 Trang strain, [9]) and for L. colombiensis. Analysis was performed on the 25 available Leishmania RNA Pol II sequences (21 existing +4 new sequences from this study), Endotrypanum monterogeii and using Trypanosoma brucei as an outgroup. The resulting Maximum Likelihood (ML) tree is shown in Fig. 5. A topologically identical Neighbour Joining (NJ) tree was also generated (Figure S2). Bootstrapping provided strong support for almost all of the nodes, the only two exceptions were 54% bootstrap support for the clade including subgenera Leishmania/Sauroleishmania/Viannia in the ML tree (Fig. 5), however, this node received 94% support in the NJ tree; and 66% support for the clade including all species except paraleishmania II in the NJ tree (Figure S2), however, this node received 85% support in the ML tree. We also tested alternate outgroups, such as Crithidia fasciculata, Leptomonas costaricensis and L. seymouri, and the topologies of the resulting ML trees were similar to Fig. 5. For example, with C. fasciculata as an outgroup the main difference was that the paraleishmania appear as a monophyletic clade, and consequently the rooting of tree is slightly different (Figure S3).

Bottom Line: Here we report a case in a 52-year-old Thai male from northern Thailand, who presented with subacute fever, huge splenomegaly and pancytopenia.This is the first report of visceral leishmaniasis caused by L. martiniquensis from the region.Moreover, the majority of parasites previously identified as "L. siamensis" also appear to be L. martiniquensis.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand.

ABSTRACT
Since 1996, there have been several case reports of autochthonous visceral leishmaniasis in Thailand. Here we report a case in a 52-year-old Thai male from northern Thailand, who presented with subacute fever, huge splenomegaly and pancytopenia. Bone marrow aspiration revealed numerous amastigotes within macrophages. Isolation of Leishmania LSCM1 into culture and DNA sequence analysis (ribosomal RNA ITS-1 and large subunit of RNA polymerase II) revealed the parasites to be members of the Leishmania enriettii complex, and apparently identical to L. martiniquensis previously reported from the Caribbean island of Martinique. This is the first report of visceral leishmaniasis caused by L. martiniquensis from the region. Moreover, the majority of parasites previously identified as "L. siamensis" also appear to be L. martiniquensis.

Show MeSH
Related in: MedlinePlus