Figure 1.
Light micrograph of Leishmania amastigotes in bone marrow aspirate.
Two infected macrophages are stained (large arrows), each with a nucleus (N) and numerous amastigotes (examples with small arrows) within the cytoplasm. The specimen was stained using Wright's stain. The bar represents 20 µm.
Figure 2.
Diagnosis of leishmaniasis by PCR.
Agarose gel electrophoresis showing PCR products using the LeF/LeR primers for Leishmania rRNA ITS-1 [17] from patient bone marrow aspirate samples compared to five Leishmania species, 1% gel, stained with ethidium bromide. Lane 1, negative control – no DNA; lane 2, L. donovani; lane 3, L. infantum; lane 4, L. tropica; lane 5, L. major; lane 6, “L. siamensis”PCM2 Trang; lane 7, BM1 is bone marrow collected 2 days after commencement of treatment; lane 8, BM2 is bone marrow collected 14 days after commencement of treatment. Size markers (MW), 100 bp ladder, are as shown.
Figure 3.
Giemsa-stained promastigote forms from culture.
A–H, examples showing morphological variation of forms observed, all at the same magnification, bar in F represents 5 µm. Procyclic-like promastigotes can be observed in A (arrows);, leptomonad-like promastigotes in B and C (arrows); nectomonad-like promastigotes in D, E and F; and rosettes and aggregates in G and H.
Figure 4.
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.
Figure 5.
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.