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BMC Infectious Diseases
Published in: BMC Infectious Diseases 1/2017

Open Access 01-12-2017 | Research article

Genetic diversity of Leishmania donovani that causes cutaneous leishmaniasis in Sri Lanka: a cross sectional study with regional comparisons

Authors: Udeshika Lakmini Kariyawasam, Angamuthu Selvapandiyan, Keshav Rai, Tasaduq Hussain Wani, Kavita Ahuja, Mizra Adil Beg, Hasitha Upendra Premathilake, Narayan Raj Bhattarai, Yamuna Deepani Siriwardena, Daibin Zhong, Guofa Zhou, Suman Rijal, Hira Nakhasi, Nadira D. Karunaweera

Published in: BMC Infectious Diseases | Issue 1/2017

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Abstract

Background

Leishmania donovani is the etiological agent of visceral leishmaniasis (VL) in the Indian subcontinent. However, it is also known to cause cutaneous leishmaniasis (CL) in Sri Lanka. Sri Lankan L. donovani differs from other L. donovani strains, both at the molecular and biochemical level. To investigate the different species or strain-specific differences of L. donovani in Sri Lanka we evaluated sequence variation of the kinetoplastid DNA (kDNA).

Methods

Parasites isolated from skin lesions of 34 CL patients and bone marrow aspirates from 4 VL patients were genotyped using the kDNA minicircle PCR analysis. A total of 301 minicircle sequences that included sequences from Sri Lanka, India, Nepal and six reference species of Leishmania were analyzed.

Results

Haplotype diversity of Sri Lankan isolates were high (H d  = 0.757) with strong inter-geographical genetic differentiation (F ST  > 0.25). In this study, L. donovani isolates clustered according to their geographic origin, while Sri Lankan isolates formed a separate cluster and were clearly distinct from other Leishmania species. Within the Sri Lankan group, there were three distinct sub-clusters formed, from CL patients who responded to standard antimony therapy, CL patients who responded poorly to antimony therapy and from VL patients. There was no specific clustering of sequences based on geographical origin within Sri Lanka.

Conclusion

This study reveals high levels of haplotype diversity of L. donovani in Sri Lanka with a distinct genetic association with clinically relevant phenotypic characteristics. The use of genetic tools to identify clinically relevant features of Leishmania parasites has important therapeutic implications for leishmaniasis.
Appendix
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Literature
1.
Alvar J, Vélez ID, Bern C, Herrero M, Desjeux P, Cano J, Jannin J, Boer M Den: Leishmaniasis worldwide and global estimates of its incidence. PLoS One 2012, 7:e35671.
2.
Imamura H, Downing T, van den Broeck F, Sanders MJ, Rijal S, Sundar S, Mannaert A, Vanaerschot M, Berg M, de Muylder G, Dumetz F, Cuypers B, Maes I, Domagalska M, Decuypere S, Rai K, Uranw S, Bhattarai NR, Khanal B, Prajapati VK, Sharma S, Stark O, Schönian G, de Koning HP, Settimo L, Vanhollebeke B, Roy S, Ostyn B, Boelaert M, Maes L, et al.: Evolutionary genomics of epidemic visceral leishmaniasis in the Indian subcontinent. elife 2016, 5(MARCH2016):1–39.
3.
WHO. Control of the leishmaniases. World Health Organ Tech Rep Ser. 2010:22–6.
4.
Kuhls K, Alam MZ, Cupolillo E, Ferreira GEM, Mauricio IL, Oddone R, Feliciangeli MD, Wirth T, Miles MA, Schönian G. Comparative microsatellite typing of new world Leishmania infantum reveals low heterogeneity among populations and its recent old world origin. PLoS Negl Trop Dis. 2011;5:1–16.CrossRef
5.
Mauricio IL, Howard MK, Stothard JR, Miles MA. Genomic diversity in the Leishmania donovani complex. Parasitology. 1999;119(3):237–46.CrossRefPubMed
6.
Pratlongl F, Bastienl P, Perello R, Lamil P, Dedetl IP, France M. Human cutaneous leishmaniasis caused by Leishmania donovani sensu stricto in Yemen. Trans R Soc Trop Med Hyg. 1995;89:398–9.CrossRef
7.
Karunaweera ND, Pratlong F, Siriwardane HVYD, Ihalamulla RL, Dedet JP. Sri Lankan cutaneous leishmaniasis is caused by Leishmania donovani zymodeme MON-37. Trans R Soc Trop Med Hyg. 2003;97:380–1.CrossRefPubMed
8.
Siriwardena HVYD, Noyes HA, Beeching NJ, Chance ML, Karunaweera ND, Bates PA. Leishmania donovani and Cutaneous leishmaniasis, Sri Lanka. Emerg Infect Dis. 2007;13:3–5.
9.
Thakur CP, Dedet JP, Narain S, Pratlong F. Leishmania species, drug unresponsiveness and visceral leishmaniasis in Bihar, India. Trans R Soc Trop Med Hyg. 2001;95:187–9.CrossRefPubMed
10.
Nawaratna SSK, Weilgama DJ, Wijekoon CJ, Dissanayake M, Rajapaksha K. Cutaneous Leishmaniasis, Sri Lanka. Emerg Infect Dis. 2007;13:1–3.CrossRef
11.
Karunaweera ND. Leishmania donovani causing cutaneous leishmaniasis in Sri Lanka: a wolf in sheep’s clothing? Trends Parasitol. 2009;25:458–63.CrossRefPubMed
12.
13.
Rajapaksa US, Ihalamulla RL, Karunaweera ND. First report of mucosal tissue localisation of leishmaniasis in Sri Lanka. Ceylon Med J. 2005;50:90–1.PubMed
14.
Rathnayake D, Ranawake RR, Sirimanna G, Siriwardhane Y, Karunaweera ND, De Silva R. Co-infection of mucosal leishmaniasis and extra pulmonary tuberculosis in a patient with inherent immune deficiency. Int J Dermatol. 2010;49:549–51.CrossRefPubMed
15.
Ranasinghe S, Zhang W-W, Wickremasinghe R, Abeygunasekera P, Chandrasekharan V, Athauda S, Mendis S, Hulangamuwa S, Matlashewski G, Pratlong F. Leishmania donovani zymodeme MON-37 isolated from an autochthonous visceral leishmaniasis patient in Sri Lanka. Pathog Glob Health. 2012;106:421–4.CrossRefPubMedPubMedCentral
16.
Refai FW, Madarasingha N, Fernandopulle R, Karunaweera ND. Nonresponsiveness to standard treatment in cutaneous leishmaniasis: a case series from Sri Lanka. Trop Parasitol. 2016;6:4–8.
17.
18.
McCall L-I, Matlashewski G. Localization and induction of the A2 virulence factor in Leishmania: evidence that A2 is a stress response protein. Mol Microbiol. 2010;77:518–30.CrossRefPubMed
19.
Zhang WW, Ramasamy G, McCall L-I, Haydock A, Ranasinghe S, Abeygunasekara P, Sirimanna G, Wickremasinghe R, Myler P, Matlashewski G. Genetic analysis of Leishmania donovani tropism using a naturally attenuated Cutaneous strain. PLoS Pathog. 2014;10:e1004244.CrossRefPubMedPubMedCentral
20.
Zhang WW, Matlashewski G. Characterization of the A2-A2rel gene cluster in Leishmania donovani: involvement of A2 in visceralization during infection. Mol Microbiol. 2001;39:935–48.CrossRefPubMed
21.
Zhang W-W. Comparison of the A2 gene locus in Leishmania donovani and Leishmania major and its control over Cutaneous infection. J Biol Chem. 2003;278:35508–15.CrossRefPubMed
22.
Beverley SM, Turco SJ. Lipophosphoglycan (LPG) and the identification of virulence genes in Leishmania. Trends Microbiol. 1998;6:35–40.CrossRefPubMed
23.
Späth GF, Epstein L, Leader B, Singer SM, Avila HA, Turco SJ, Beverley SM. Lipophosphoglycan is a virulence factor distinct from related glycoconjugates in the protozoan parasite Leishmania major. Proc Natl Acad Sci U S A. 2000;97:9258–63.CrossRefPubMedPubMedCentral
24.
Turco SJ, Späth GF, Beverley SM. Is lipophosphoglycan a virulence factor? A surprising diversity between Leishmania species. Trends Parasitol. 2001;17:223–6.CrossRefPubMed
25.
26.
Chen DQ, Kolli BK, Yadava N, Lu HG, Gilman-Sachs A, Peterson DA, Chang KP. Episomal expression of specific sense and antisense mRNAs in Leishmania amazonensis: modulation of gp63 level in promastigotes and their infection of macrophages in vitro. Infect Immun. 2000;68:80–6.CrossRefPubMedPubMedCentral
27.
Downing T, Imamura H, Decuypere S, Clark TG, Coombs GH, Cotton JA, Hilley JD, de Doncker S, Maes I, Mottram JC, Quail MA, Rijal S, Sanders M, Schonian G, Stark O, Sundar S, Vanaerschot M, Hertz-Fowler C, Dujardin J-C, Berriman M. Whole genome sequencing of multiple Leishmania donovani clinical isolates provides insights into population structure and mechanisms of drug resistance. Genome Res. 2011;21:2143–56.CrossRefPubMedPubMedCentral
28.
Morales MA, Chicharro C, Ares M, Cañavate C, Barker DC, Alvar J. Molecular tracking of infections by Leishmania infantum. Trans R Soc Trop Med Hyg. 2001;95:104–7.CrossRefPubMed
29.
Peacock CS, Seeger K, Harris D, Murphy L, Ruiz JC, Quail MA, Peters N, Adlem E, Tivey A, Aslett M, Kerhornou A, Ivens A, Fraser A, Rajandream M-A, Carver T, Norbertczak H, Chillingworth T, Hance Z, Jagels K, Moule S, Ormond D, Rutter S, Squares R, Whitehead S, Rabbinowitsch E, Arrowsmith C, White B, Thurston S, Bringaud F, Baldauf SL, et al. Comparative genomic analysis of three Leishmania species that cause diverse human disease. Nat Genet. 2007;39:839–47.CrossRefPubMedPubMedCentral
30.
31.
Zemanová E, Jirků M, Mauricio IL, Miles MA, Lukes J. Genetic polymorphism within the Leishmania donovani complex: correlation with geographic origin. Am J Trop Med Hyg. 2004;70:613–7.PubMed
32.
Selvapandiyan A, Duncan R, Mendez J, Kumar R, Salotra P, Cardo LJ, Nakhasi HL. A Leishmania minicircle DNA footprint assay for sensitive detection and rapid speciation of clinical isolates. Transfusion. 2008;48:1787–98.CrossRefPubMed
33.
Singh N, Curran MD, Middleton D, Rastogi AK. Characterization of kinetoplast DNA minicircles of an Indian isolate of Leishmania donovani. Acta Trop. 1999;73:313–9.CrossRefPubMed
34.
Singh N, Curran MD, Rastogil AK, Middleton D, Sundar S. Diagnostic PCR with Leishmania donovani specificity using sequences from the variable region of kinetoplast minicircle DNA. Trop Med Int Heal. 1999;4:448–53.CrossRef
35.
Lukes J, Mauricio IL, Schönian G, Dujardin J-C, Soteriadou K, Dedet J-P, Kuhls K, Tintaya KWQ, Jirků M, Chocholová E, Haralambous C, Pratlong F, Oborník M, Horák A, Ayala FJ, Miles MA. Evolutionary and geographical history of the Leishmania donovani complex with a revision of current taxonomy. Proc Natl Acad Sci U S A. 2007;104:9375–80.CrossRefPubMedPubMedCentral
36.
Liu B, Liu Y, Motyka SA, Agbo EEC, Englund PT. Fellowship of the rings: the replication of kinetoplast DNA. Trends Parasitol. 2005;21:363–9.CrossRefPubMed
37.
38.
Reithinger R, Dujardin JC. Molecular diagnosis of leishmaniasis: Current status and future applications. J Clin Microbiol. 2007;45(1):21–5.
39.
Safi N. Evaluation of thermotherapy for the treatment of cutaneous Leishmaniasis in Kabul, Afghanistan: a randomized controlled trial. Int J Infect Dis. 2012;16(March):e174.CrossRef
40.
Silva SY, Rueda LC, López M, Vélez ID, Rueda-Clausen CF, Smith DJ, Muñoz G, Mosquera H, Silva FA, Buitrago A, Díaz H, López-Jaramillo P. Double blind, randomized controlled trial, to evaluate the effectiveness of a controlled nitric oxide releasing patch versus meglumine antimoniate in the treatment of cutaneous leishmaniasis [NCT00317629]. Trials. 2006;7:14.CrossRefPubMedPubMedCentral
41.
Batista LFDS, Segatto M, Guedes CES, Sousa RS, Rodrigues CAT, JCM B, Silva JS, Santos SO, Larangeira D, Macedo AM, Schriefer A, PST V. An assessment of the genetic diversity of Leishmania infantum isolates from infected dogs in Brazil. Am J Trop Med Hyg. 2012;86:799–806.CrossRefPubMed
42.
Ranasinghe S, Wickremasinghe R, Hulangamuwa S, Sirimanna G, Opathella N, Maingon RDC, Chandrasekharan V. Polymerase chain reaction detection of Leishmania DNA in skin biopsy samples in Sri Lanka where the causative agent of cutaneous leishmaniasis is Leishmania donovani. Mem Inst Oswaldo Cruz. 2015;110:1017–23.CrossRefPubMedPubMedCentral
43.
Tsokana CN, Athanasiou LV, Valiakos G, Spyrou V, Manolakou K, Billinis C. Molecular Diagnosis of Leishmaniasis, Species Identification and Phylogenetic Analysis, Leishmaniasis - Trends in Epidemiology, Diagnosis and Treatment, Dr. David Claborn (Ed.), Chapter 8; 2014. p. 161–193.
44.
Thompson JD, Higgins DG, Gibson TJ. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 1994;22:4673–80.CrossRefPubMedPubMedCentral
45.
Larkin MA, Blackshields G, Brown NP, Chenna R, Mcgettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DG. Clustal W and Clustal X version 2.0. Bioinformatics. 2007;23:2947–8.CrossRefPubMed
46.
Librado P, Rozas J. DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics. 2009;25:1451–2.CrossRefPubMed
47.
Tajima F. Statistical method for testing the neutral mutation hypothesis by DNA polymorphism. Genetics. 1989;123:585–95.PubMedPubMedCentral
48.
Fu YX. Statistical tests of neutrality of mutations against population growth, hitchhiking and background selection. Genetics. 1997;147:915–25.PubMedPubMedCentral
49.
Ashfaq M, Hebert PDN, Mirza MS, Khan AM, Mansoor S, Shah GS, Zafar Y. DNA barcoding of Bemisia Tabaci Complex (Hemiptera: Aleyrodidae) reveals southerly expansion of the dominant whitefly species on cotton in Pakistan. PLoS One. 2014;9:1–14.
50.
Goodall-Copestake WP, Tarling GA, Murphy EJ. On the comparison of population-level estimates of haplotype and nucleotide diversity: a case study using the gene cox1 in animals. Heredity (Edinb). 2012;109:50–6.CrossRef
51.
Excoffier L, Laval G, Schneider S. Arlequin (version 3.0): an integrated software package for population genetics data analysis. Evol Bioinformatics Online. 2005;1:47–50.
52.
Wright S. Variability Within and Among Natural Populations. Evolution and the Genetics of Populations, Volume 4. University of Chicago Press, USA; 1978.
53.
54.
55.
Ramírez JD, Hernández C, León CM, Ayala MS, Flórez C, González C. Taxonomy, diversity, temporal and geographical distribution of Cutaneous Leishmaniasis in Colombia: A retrospective study. Scientific Reports. 2016;6(1).
56.
Bhattarai NR, Dujardin JC, Rijal S, De Doncker S, Boelaert M, Van der Auwera G. Development and evaluation of different PCR-based typing methods for discrimination of Leishmania donovani isolates from Nepal. Parasitology. 2010;137:947–57.CrossRefPubMed
57.
Pritchard JK, Stephens M, Donnelly P. Inference of population structure using multilocus genotype data. Genetics. 2000;155:945–59.PubMedPubMedCentral
58.
Alonso DP, Costa DL, Mendonça IL, De Henrique C, Costa N, Eduardo P, Ribolla M. Short report: heterogeneity of Leishmania infantum chagasi Kinetoplast DNA in Teresina (Brazil). Am J Trop Med Hyg. 2010;82:819–21.CrossRefPubMedPubMedCentral
59.
de Oliveira JP, Fernandes F, Cruz AK, Trombela V, Monteiro E, Camargo AA, Barral A, de Oliveira CI. Genetic diversity of Leishmania amazonensis strains isolated in northeastern Brazil as revealed by DNA sequencing, PCR-based analyses and molecular karyotyping. Kinetoplastid Biol Dis. 2007;6(Lcl):5.CrossRefPubMedPubMedCentral
60.
Seblova V, Myskova J, Hlavacova J, Votypka J, Antoniou M, Volf P. Natural hybrid of Leishmania infantum/L. donovani: development in Phlebotomus tobbi, P. perniciosus and Lutzomyia longipalpis and comparison with non-hybrid strains differing in tissue tropism. Parasit Vectors. 2015;8:605.CrossRefPubMedPubMedCentral
61.
Segatto M, Ribeiro LS, Costa DL, Costa CHN, de Oliveira MR, Carvalho SFG, Macedo AM, Valadares HMS, Dietze R, de Brito CFA, Lemos EM. Genetic diversity of Leishmania infantum field populations from Brazil. Mem Inst Oswaldo Cruz. 2012;107:39–47.CrossRefPubMed
62.
Peters NC, Egen JG, Secundino N, Debrabant A, Kimblin N, Kamhawi S, Lawyer P, Fay MP, Germain RN, Sacks D. Demonstration of genetic exchange during cyclical development of Leishmania in the sand fly vector. Science (80-). 2009;324:265–8.CrossRef
63.
Gajapathy K, Jude PJ, Surendran SN. Morphometric and meristic characterization of Phlebotomus argentipes species complex in northern Sri Lanka: evidence for the presence of potential leishmaniasis vectors in the country. Trop Biomed. 2011;28:259–68.PubMed
64.
Ozbel Y, Sanjoba C, Alten B, Asada M, Depaquit J, Matsumoto Y, Demir S, RRMLR S, RPVJ R, Matsumoto Y. Distribution and ecological aspects of sand fly (Diptera: Psychodidae) species in Sri Lanka. J Vector Ecol. 2011;36 Suppl 1(Karunaweera):S77–86.CrossRefPubMed
65.
Karunaweera,N.D., Siriwardana HVYD, Karunanayake P.: Management of leishmaniasis. Sri Lanka Prescr 2013, 21:1–5.
66.
Wijesundera MS. Cutaneous leishmaniasis: an emerging health risk in Sri Lanka. Ceylon Med J. 2001;46:151–2.CrossRefPubMed
67.
Naotunne TD, Rajakulendran S, Abeyewickreme W, Kulasiri CD, Perera J, Premaratne U. Cutaneous leishmaniasis in Sri Lanka: an imported disease linked to the middle-east and African employment boom. Trop Geogr Med. 1990;42:72–4.PubMed
68.
Dissanayake AS: Parasitic zoonoses with special reference to Sri Lanka. In In: Elizabeth U canning Ed. parasitological topics. Society of Protozoologists, Kansas USA: Allen Press Inc., 1981: 78–85.
69.
Sang DK, Njeru WK, Ashford RW. A possible animal reservoir for Leishmania tropica s.L. in Kenya. Ann Trop Med Parasitol. 1992;86:311–2.CrossRefPubMed
Metadata
Title
Genetic diversity of Leishmania donovani that causes cutaneous leishmaniasis in Sri Lanka: a cross sectional study with regional comparisons
Authors
Udeshika Lakmini Kariyawasam
Angamuthu Selvapandiyan
Keshav Rai
Tasaduq Hussain Wani
Kavita Ahuja
Mizra Adil Beg
Hasitha Upendra Premathilake
Narayan Raj Bhattarai
Yamuna Deepani Siriwardena
Daibin Zhong
Guofa Zhou
Suman Rijal
Hira Nakhasi
Nadira D. Karunaweera
Publication date
01-12-2017
Publisher
BioMed Central
Published in
BMC Infectious Diseases / Issue 1/2017
Electronic ISSN: 1471-2334
DOI
https://doi.org/10.1186/s12879-017-2883-x

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