Molecular detection and conventional identification of leishmania species in reservoir hosts of zoonotic cutaneous leishmaniasis in fars province, South of iran.
AbstractBackground: The objectives of our research were to search for Leishmania species in rodents in Fars province, south of Iran, and to compare molecular with conventional methods for detecting these para-sites.Methods: Rodents were captured using live traps and screened for Leishmania species using molecular and conventional methods, including the taking of smears from each ear. Nested PCR was employed to detect Leishmania in rodents by amplifying a region of the ribosomal RNA amplicon of Leishmania (ITS1- 5.8S rRNA-ITS2) that is species-specific by DNA sequence.Results: Totally, 122 rodents were captured. Leishmania parasites were detected using the nested PCR and three conventional methods (direct smear, NNN culture and Balb/C inoculation. 41 (33.6%) out of 122 rodents had Leishmania infections (34 Meriones lybicus and 7 M. persicus). All PCR products of the ITS-rDNA gene were sequenced. Sequence analysis revealed that 28 out of 41 positive samples were Leishma-nia major. Thirteen sequences were unreadable and therefore not identified.Conclusion: At least two gerbil species common in Fars ZCL foci, M. lybicus and M. persicus, are acquir-ing infections of L. major and may be reservoir hosts of one predominant parasite haplotype. Most infec-tions were detected molecularly not by conventional methods, because most rodents died in the traps.
Alvar J, Vйlez ID, Bern C, Herrero M, Desjeux P, Cano J, Jannin J, den Boer M. Leishmaniasis worldwide and global estimates of its incidence. PLoS ONE. 2012; 7: e35671.
Parvizi P, Ready PD. Nested PCRs of nuclear ITS-rDNA fragments detect three Leishmania species of gerbils in sandflies from Iranian foci of zoonotic cutaneous leishmaniasis. Trop Med Int Health. 2008; 13:1159-71.
Mirzaei A, Rouhani S, Taherkhani H, Farahmand M, Kazemi B, Hedayati M, Davari B, Parvizi P. Isolation and detection of Leishmania species among naturally infected Rhombomis opimus, a reservoir host of zoonotic cutaneous leishmaniasis in Turkemen Sahara, North East of Iran. Exp Parasitol. 2011; 129: 378-380.
Parvizi P, Alaeenovin E, Kazerooni PA, Ready PD. Low diversity of Leishmania para-sites in sandflies and the absence of the great gerbil in foci of zoonotic cutaneous leishmaniasis in Fars province, southern Iran. Trans R Soc Trop Med Hyg. 2013; in press.
WHO. Control of the Leishmaniasis. Tech-nical Report Series No.793; 1990; World Health Organization, Geneva.
WHO. A new edition of a WHO book - 'Technical Report Series on the Control of Leishmaniasis', 2013; World Health Organiza-tion, Geneva. 7. Ready PD. Biology of phlebotomine sandflies as vectors of disease agents. Annu Rev Entomol. 2013; 58: 227-50. 8. Azizi K, Rassi Y, Javadian E, Motazedian MH, Rafizadeh S, Yaghoobi-Ershadi MR, Mohebali M. Phlebotomus (Paraphlebotomus) alexandri: a prob-able vector of Leishmania infantum in Iran. Ann Trop Med Parasitol. 2006; 100: 63-8.
Rassi Y, Javadian E, Amin M, Rafizadeh S, Vatandoost H, Motazedian H. Meriones libycus is the main reservoir of zoonotic cutaneous leishmaniasis in south Islamic Republic of Iran. East Mediterr Health J. 2006; 12: 474-477.
Bensoussan E, Nasereddin A, Jonas F, Schnur LF, Jaffe CL., Comparison of PCR Assays for Diagnosis of Cutaneous Leishmaniasis. J Clin Microbiol. 2006; 44: 1435–1439.
Nadim A, Faghih M. 1968. The epidemiology of cutaneous leishmaniasis in the Isfahan province of Iran: I. The reservoir II. The hu-man disease. Trans R Soc Trop Med Hyg. 1968; 62: 534-42.
Nadim A, Seyedi-Rashti MA. A brief review of the epidemiology of various types of leishmaniasis in Iran. Acta Med Iran. 1971; 14: 99-106. 13. Edrissian GH. Visceral leishmaniasis in Iran and the role of serological tests in diagnosis and epidemiological studies. In: Ali Ozel, M., Ziyal lkass, M. (Ed.), Parasitology for the Twenty First Century. CAB Int. 1996; pp. 63-78.
Ardehali S, Moattari A, Hatam GR, Hosseini SMH, Sharifi I. Characterization of Leishmania isolate in Iran: serotyping with species specific monoclonal antibodies. Acta Trop. 2000; 75: 301–307.
Parvizi P, Moradi G, Akbari G, Ready PD, Farahmand M, Piazak N, Assmar M, Amirkhani A. PCR Detection and sequencing of parasite ITS-rDNA gene from reservoirs host of zoonotic cutaneous leishmaniasis in central of Iran. Parasitol Res. 2008; 103: 1273- 1278.
Rassi Y, Jalali M, Javadian E, Moatazedian MH. Confirmation of Meriones libycus (Rodentia; Gerbillidae) as the Main Reservoir Host of Zoonotic Cutaneous Leishmaniasis in Arsanjan, Fars Province, South of Iran (1999-2000). Iranian J Publ Health. 2001; 30: 143-144.
Schцnian G, Kuhls K, Mauricio I. Molecular approaches for a better understanding of the epidemiology and population genetics of Leishmania. Parasitology. 2011; 138: 405-25.
Talmi-Frank D, Nasereddin A, Schnur LF, Schonian G, Toz SO, Jaffe CL, Baneth G. Detection and Identification of Old World Leishmania by High Resolution Melt Analysis. Plos Negl Trop Dis. 2010; 4: e581.
Akhavan AA, Yaghoobi-Ershadi MR, Kha-mesipour A, Mirhendi H, Alimohammad-ian MH, Rassi Y, Arandian MH, Jafari R, Abdoli H, Shareghi N, Ghanei M, Jalali-zand N. Dynam-ics of Leishmania infection rates in Rhombomys opimus (Rodentia: Gerbillinae) population of an endemic focus of zoonotic cutaneous leishmaniasis in Iran. Bull Soc Pathol Exot. 2010; 103: 84-89.
Mohebali M, Javadian E, Yaghoobi-Ershadi M, Akhavan A, Hajjaran H, Abaei M. Charac-terization of Leishmania infection in rodents from endemic areas of the Islamic Republic of Iran. East Mediterr Health J. 2004; 10: 591-9.
Parvizi P, Mauricio I, Aransay AM, Miles MA, Ready PD. First detection of Leishmania major in peridomestic Iranian sandflies: comparison of nested PCR of nuclear ITS ribosomal DNA and semi-nested PCR of minicircle kinetoplast DNA. Acta Trop. 2005; 93: 75-83.
Cupolillo E, Grimaldi JG, Momen H, Beverly SM. Intergenic region typing (IRT): A rapid molecular approach to the characterization and evolution of Leishmania. Mol Bioch Parasitol. 1995; 73: 145-155.
Mahmoudzadeh-Niknam H, Ajdary S, Riazi-Rad F, Mirzadegan E, Rezaeian A, Khaze V, Djadid ND, Alimohammadian MH. Molecu-lar epidemiology of cutaneous leishmaniasis and heterogeneity of Leishmania major strains in Iran. Trop Med Int Health. 2012; doi: 10.1111/j.1365-3156.2012.03078.x. In press.
Ziaei H. A field guide for identifying of Iranian desert mammalians. Tehran, Iran, Iranian En-vironment Organization. 1996.
Boitani L, Bartoli S. Macdonald encyclopedia of mammals, 1st ed. London: Macdonald & Co. 1980
Strelkova MV, Eliseev LN, Ponirovsky EN, Dergacheva TI, Evans DA. Mixed leishmanial infections in Rhombomys optimus: a key to the persistence of Leishmania major from one transmission season to the next. Ann Trop Med Parasitol. 2001; 95: 811-819. 27. Yaghoobi-Ershadi MR, Javadian E, Tahvildare-Bidruni GH. Leishmania major MON-26 isolated from naturally infected Phlebotomus papatasi (Diptera: Psychodidae) in Isfahan province, Iran. Acta Trop. 1995; 59: 279–282.
Akhavan AA, Ghods R, Jeddi-Tehrani M, Yaghoobi-Ershadi MR, Khamesipour A, Mahmoudi AR. Production and Purification of Anti-Rhombomys opimus Immunoglobulins. Iran J Arthropod-Borne Dis. 2011; 5: 69-76. 29. Momenbella Fard MD, Kalantari M, Rassi Y, Javadian E. The PCR based detection of Leishmania major infection in Meriones libycus (rodentia:Muridae) from soutern Iran. Ann Trop. Med Parasitol. 2003; 79: 811-816.
Asgari Q, Motazedian MH, Mehrabani D, Oryan A, Hatam GR, Owji SM, Oryan A. 2007. Zoonotic cutaneous leishmaniasis in Shiraz, Southern Iran: A molecular, isoenzyme and morphologic approach. J Res Med Sci. 2007; 12: 7-15.
Yaghoobi-Ershadi MR, Akhavan AA, Mohebali M. Meriones libycus and Rhombomys opiums(Rodentia: Gerbillidae) are the main res-ervoir hosts in e new focus of zoonotic Cuta-neous leishmaniasis in Iran. Trans R Soc Trop Med Hyg. 1996; 90: 503-504.
Rassi Y, Oshaghi MA, MohammadiAzani S, Abaei MR, Rafizadeh S, Mohebali M, Mohtarami F, Zainali MK. Molecular detec-tion of Leishmania infection due to leishmania major and Leishmania turanica in the vectors and reservoir host in Iran. Vector-Borne and Zo-onotic Dis. 2011; 11: 145-150.
Edrissian GH, Ghorbani M, Tahvildar-Bidruni G. Meriones persicus, another proba-ble reservoir of zoonotic cutaneous leishmaniasis in Iran. Trans R Soc Trop Med Hyg. 1975; 69: 517–9.
Mohebali M, Poormohammadi B, Kanani A, Hajjaran, Edrissian GH. Rodents: another group of animal reservoir hosts of visceral leishmaniasis in Meshkin-Shahr district, the Is-lamic Republic of