Molecular analysis and infection Determination of visceral leishmaniasis in asymptomatic dogs in Meshkin-Shahr Ardabil province, northwestern Iran

Subject Areas : Journal of Comparative Pathobiology

روزبه Taslimian ¹ ( Department of Pathobiology, Science and Research branch, Islamic Azad University, Tehran, Iran )
بهار Shemshadi ² ( Department of Pathobiology, Science and Research branch, Islamic Azad University, Tehran, Iran )
عادل Spotin ³ ( Immunology Research Centre, Tabriz University of Medical Sciences, Tabriz, Iran )
R. Fotouhi Ardakani ⁴ ( Cellular and Molecular Research Centre, Qom University of Medical Science, Qom, Iran )
نرمین najfzadeh ⁵ ( pasture institute, tehran, Iran )

Received: 2021-11-03 Accepted : 2021-11-03 Published : 2020-12-21

Keywords: Asymptomatic dogs, Visceral Leishmaniasis, non-invasive method, PCR, Ardabil,

Abstract :

Leishmania infantum parasite is the main causative agent of visceral leishmaniasis (VL) which threatens a wide range of humans and dogs in Iran. Our objective was to investigate Leishmania parasite species in Ardebil Province a well-known VL foci targeting ITS-rDNA gene, sequencing and phylogenetic analysis. Vacuum EDTA tubes were used for blood collection of asymptomatic dogs using non-invasive methods in Ardabil, northwestern Iran in August 2019. Phenol-chloroform DNA extraction, ITS-rDNA gene PCR and sequencing were performed. Leishmania species was accurately identified based on sequences and Maximum likelihood phylogenic analyzes. 11 of 31 asymptomatic dogs were definitively infected with L. infantum. Under 5 years old age group had the highest infection rate, which showed that this group is more sensitive to VL in this area and also the infection was mainly observed in male sheepdogs. These findings show that non-invasive sampling and molecular methods are reliable and convenient in the diagnosis of visceral leishmaniasis. Regarding the 35.5% infection ratio in the area control and monitoring measures of human individuals and canine reservoirs should be taken to consideration.

References:

1. Taslimian R, Shemshadi B, Spotin A, Ardakani RF, Parvizi P. Molecular Characterization of Visceral Leishmaniasis in Asymptomatic Dogs in North Khorasan, Northeastern Iran. Jundishapur Journal of Microbiology. 2019;12(12):1-8.


2. Mohebali M, Moradi-Asl E, Rassi Y. Geographic distribution and spatial analysis of Leishmania infantum infection in domestic and wild animal reservoir hosts of zoonotic visceral leishmaniasis in Iran: A systematic review. Journal of vector borne diseases. 2018;55(3):173-83.


3. Hajjaran H, Mohebali M, Mamishi S, Vasigheh F, Oshaghi MA, Naddaf SR, et al. Molecular identification and polymorphismdetermination of cutaneous and visceral leishmaniasis agents isolated from human and animal hosts in Iran. BioMed research international. 2013;2013.

4. Travi BL, Cordeiro-da-Silva A, Dantas-Torres F, Miro G. Canine visceral leishmaniasis: diagnosis and management of the reservoir living among us. PLoS neglected tropical diseases. 2018;12(1):e0006082.


5. Monteiro FM, Machado AS, Rocha-Silva F, Assunção CB, Graciele-Melo C, Costa LE, et al. Canine visceral leishmaniasis: detection of Leishmania spp. genome in peripheral blood of seropositive dogs by real-time polymerase chain reaction (rt-PCR). Microbial pathogenesis. 2019;126:263-8.


6. Mody RM, Lakhal-Naouar I, Sherwood JE, Koles NL, Shaw D, Bigley DP, et al. Asymptomatic Visceral Leishmania infantum Infection in US Soldiers Deployed to Iraq. Clinical Infectious Diseases. 2018;68(12):2036-44.


7. Abdinia B, Oliaei-Motlagh M, Teimouri-Dereshki A. Pediatric visceral leishmaniasis in northwest of Iran. Medicine. 2016;95(44):e5261.


8. Velez R, Ballart C, Domenech E, Abras A, Fernández-Arévalo A, Gómez S, et al. Seroprevalence of canine Leishmania infantum infection in the Mediterranean region and identification of risk factors: The example of North-Eastern and Pyrenean areas of Spain. Preventive veterinary medicine. 2019;162:67-75.


9. Abbaszadeh-Afshar Mj, Sharifi I, Bamorovat M, Mohebali M, Bahreini MS, Naderi A. Canine Visceral Leishmaniasis; A Seroepidemiological Survey in Jiroft District, Southern Kerman Province, Southeastern Iran in 2015. Iranian journal of parasitology. 2018;13(1):67-71.


10. Baneth G, Koutinas AF, Solano-Gallego L, Bourdeau P, Ferrer L. Canine leishmaniosis–new concepts and insights on an expanding zoonosis: part one. Trends in parasitology. 2008;24(7):324-30.


11. Herrera G, Higuera A, Patiño LH, Ayala MS, Ramírez JD. Description of Leishmania species among dogs and humans in Colombian Visceral Leishmaniasis outbreaks. Infection, Genetics and Evolution. 2018;64:135-8.


12. Teimouri A, Mohebali M, Kazemirad E, Hajjaran H. Molecular Identification of Agents of Human Cutaneous Leishmaniasis and Canine Visceral Leishmaniasis in Different Areas of Iran Using Internal Transcribed Spacer 1 PCR-RFLP. Journal of arthropod-borne diseases. 2018;12(2):162-71.


13. Aschar M, de Oliveira ETB, Laurenti MD, Marcondes M, Tolezano JE, Hiramoto RM, et al. Value of the oral swab for the molecular diagnosis of dogs in different stages of infection with Leishmania infantum. Veterinary parasitology. 2016;225:108-13.


14. Helsinki. World Medical Association Declaration of Helsinki. Ethical principles for medical research involving human subjects. Bulletin of the World Health Organization. 2001;79(4):373.


15. Parvizi P, Mauricio I, Aransay A, Miles M, Ready P. First detection of Leishmania major in peridomestic Phlebotomus papatasi from Isfahan province, Iran: comparison of nested PCR of nuclear ITS ribosomal DNA and semi-nested PCR of minicircle kinetoplast DNA. Acta tropica. 2005;93(1):75-83.


16. Parvizi P, Ready P. Nested PCRs and sequencing of nuclear ITS‐rDNA fragments detect three Leishmania species of gerbils in sandflies from Iranian foci of zoonotic cutaneous leishmaniasis. Tropical Medicine & International Health. 2008;13(9):1159-71.


17. Mohebali M, Arzamani K, Zarei Z, Akhoundi B, Hajjaran H, Raeghi S, et al. Canine visceral leishmaniasis in wild canines (fox, jackal, and wolf) in northeastern Iran using parasitological, serological, and molecular methods. Journal of arthropod-borne diseases. 2016;10(4):538-45. 28032106.


18. Leite RS, de Almeida Ferreira S, Ituassu LT, de Melo MN, de Andrade ASR. PCR diagnosis of visceral leishmaniasis in asymptomatic dogs using conjunctival swab samples. Veterinary parasitology. 2010;170(3-4):201-6.


19. Alvar J, Canavate C, Molina R, Moreno J, Nieto J. Canine leishmaniasis. Advances in parasitology. 2004;57(3):1-88.


20. Santos-Oliveira JR, Da-Cruz AM, Pires LH, Cupolillo E, Kuhls K, Giacoia-Gripp CB, et al. Atypical Lesions as a Sign of Cutaneous Dissemination of Visceral Leishmaniasis in a Human Immunodeficiency Virus–Positive Patient Simultaneously Infected by Two Viscerotropic Leishmania Species. The American journal of tropical medicine and hygiene. 2011;85(1):55-9.


21. Baneth G, Yasur-Landau D, Gilad M, Nachum-Biala Y. Canine leishmaniosis caused by Leishmania major and Leishmania tropica: comparative findings and serology. Parasites & vectors. 2017;10(1):113.


22. Costa DN, Codeço CT, Silva MA, Werneck GL. Culling dogs in scenarios of imperfect control: realistic impact on the prevalence of canine visceral leishmaniasis. PLoS neglected tropical diseases. 2013;7(8):e2355.


23. Courtenay O, Bazmani A, Parvizi P, Ready PD, Cameron MM. Insecticide–impregnated dog collars reduce infantile clinical visceral leishmaniasis under operational conditions in NW Iran: A community–wide cluster randomised trial. PLoS neglected tropical diseases. 2019;13(3):e0007193.

24. Molaei S, Dalimi A, Mohebali M, Zareii Z, Mohamadi B, Akhondi B et al . Study of Canine Visceral Leishmaniasis in Symptomatic and Asymptomatic Domestic Dogs in Meshkinshahr City, Iran. J Ardabil Univ Med Sci. 2016; 16 (1) :105-115