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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.

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Related in: MedlinePlus

Phylogenetic analysis of Leishmania strain CM1 using ITS-1 sequences.A: ML tree including LSCM1 (accession number JX898938) with various “L. siamensis” from Thailand and elsewhere: GQ281278 is from a horse, Germany [11]; GQ281282 is from a cow, Switzerland [12]; JX195637 is of human origin from Stun, Thailand [29]; JQ617283 is from a horse, USA [13]; JQ001751, JQ001752 are human isolates from Trang and Songkhla, Thailand [10]; GQ226034 is of human origin from Chantaburi, Thailand [7]; EF200012 is of human origin from Phang-nga, Thailand [5]; JX195640 is the human PCM2 isolate from Trang, Thailand [9]. Also included are sequences from L. enriettii, L. infantum, L. martiniquensis and Leishmania from Australia (AM-2004). B: NJ tree on the same dataset. Numbers at nodes indicate bootstrap values on 1000 replicates and L. infantum was used as an outgroup.
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pntd-0003339-g004: Phylogenetic analysis of Leishmania strain CM1 using ITS-1 sequences.A: ML tree including LSCM1 (accession number JX898938) with various “L. siamensis” from Thailand and elsewhere: GQ281278 is from a horse, Germany [11]; GQ281282 is from a cow, Switzerland [12]; JX195637 is of human origin from Stun, Thailand [29]; JQ617283 is from a horse, USA [13]; JQ001751, JQ001752 are human isolates from Trang and Songkhla, Thailand [10]; GQ226034 is of human origin from Chantaburi, Thailand [7]; EF200012 is of human origin from Phang-nga, Thailand [5]; JX195640 is the human PCM2 isolate from Trang, Thailand [9]. Also included are sequences from L. enriettii, L. infantum, L. martiniquensis and Leishmania from Australia (AM-2004). B: NJ tree on the same dataset. Numbers at nodes indicate bootstrap values on 1000 replicates and L. infantum was used as an outgroup.

Mentions: An initial molecular identification of LSCM1 was performed by cloning and sequencing the PCR product of the LeF/LeR primers. This was performed on DNA extracted from both an initial BM sample taken on the first day of admission to hospital (before treatment) and on the subsequent culture derived from the BM aspirate. The resulting ITS-1 DNA sequences were identical to each other, and also were identical or very similar to several of the nucleotide sequences previously reported for “L. siamensis” (Figure S1A), but differed from another “L. siamensis”, the PCM2 Trang strain (Figure S1B). However, the LSCM1 ITS-1 sequence was also identical to that of L. martiniquensis (Fig. 4A, 4B). These results indicate that LSCM1 is L. martiniquensis, and further suggest that several other so-called “L. siamensis” isolates may in fact be L. martiniquensis. The other member of the L. enriettii complex shown in Fig. 4 is the parasite described by Dougall et al. [16], a Leishmania species from Australia infecting kangaroos that also has not yet been formally named (AM-2004).


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 of Leishmania strain CM1 using ITS-1 sequences.A: ML tree including LSCM1 (accession number JX898938) with various “L. siamensis” from Thailand and elsewhere: GQ281278 is from a horse, Germany [11]; GQ281282 is from a cow, Switzerland [12]; JX195637 is of human origin from Stun, Thailand [29]; JQ617283 is from a horse, USA [13]; JQ001751, JQ001752 are human isolates from Trang and Songkhla, Thailand [10]; GQ226034 is of human origin from Chantaburi, Thailand [7]; EF200012 is of human origin from Phang-nga, Thailand [5]; JX195640 is the human PCM2 isolate from Trang, Thailand [9]. Also included are sequences from L. enriettii, L. infantum, L. martiniquensis and Leishmania from Australia (AM-2004). B: NJ tree on the same dataset. Numbers at nodes indicate bootstrap values on 1000 replicates and L. infantum was used as an outgroup.
© Copyright Policy
Related In: Results  -  Collection

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

pntd-0003339-g004: Phylogenetic analysis of Leishmania strain CM1 using ITS-1 sequences.A: ML tree including LSCM1 (accession number JX898938) with various “L. siamensis” from Thailand and elsewhere: GQ281278 is from a horse, Germany [11]; GQ281282 is from a cow, Switzerland [12]; JX195637 is of human origin from Stun, Thailand [29]; JQ617283 is from a horse, USA [13]; JQ001751, JQ001752 are human isolates from Trang and Songkhla, Thailand [10]; GQ226034 is of human origin from Chantaburi, Thailand [7]; EF200012 is of human origin from Phang-nga, Thailand [5]; JX195640 is the human PCM2 isolate from Trang, Thailand [9]. Also included are sequences from L. enriettii, L. infantum, L. martiniquensis and Leishmania from Australia (AM-2004). B: NJ tree on the same dataset. Numbers at nodes indicate bootstrap values on 1000 replicates and L. infantum was used as an outgroup.
Mentions: An initial molecular identification of LSCM1 was performed by cloning and sequencing the PCR product of the LeF/LeR primers. This was performed on DNA extracted from both an initial BM sample taken on the first day of admission to hospital (before treatment) and on the subsequent culture derived from the BM aspirate. The resulting ITS-1 DNA sequences were identical to each other, and also were identical or very similar to several of the nucleotide sequences previously reported for “L. siamensis” (Figure S1A), but differed from another “L. siamensis”, the PCM2 Trang strain (Figure S1B). However, the LSCM1 ITS-1 sequence was also identical to that of L. martiniquensis (Fig. 4A, 4B). These results indicate that LSCM1 is L. martiniquensis, and further suggest that several other so-called “L. siamensis” isolates may in fact be L. martiniquensis. The other member of the L. enriettii complex shown in Fig. 4 is the parasite described by Dougall et al. [16], a Leishmania species from Australia infecting kangaroos that also has not yet been formally named (AM-2004).

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