Morphological Description, Phylogenetic and Molecular Analysis of Dirofilaria immitis Isolated from Dogs in the Northwest of Iran
Background: Dirofilariasis is a globally distributed arthropod-borne parasitic disease of mainly canids and felids. We evaluated to extend the knowledge of morpho-molecular characteristics and outer ultrastructure of Dirofilaria immitis isolated from Northwest of Iran.
Methods: Overall, 67 filarial worms including 41 females and 26 males parasites were collected from the cardiovascular system of the 43 stray dogs in Meshkinshar, Ardebil Province, Northwest of Iran in 2017, and subjected to light and scanning electron microscopy (SEM) as well as carmine alum staining for morpho-molecular and identification. Molecular methods were used for confirmation of morphological findings by sequencing of Cytochrome c oxidase subunit I (cox1) gene.
Results: The partial DNA sequencing of cox1 gene of adult parasites showed considerable homology and close proximity to the previously isolated from Kerman and Meshkinshahr, Iran. The lowest genetic variation and the highest intra-species variability was found in D. immitis and Dirofilaria repens, respectively. No similarity was identified between D. immitis nucleotide sequence and Wolbachia species as its endosymbiont bacteria.
Conclusion: The SEM technique is an excellent tool for differential recognition of the parasite surface morphology and molecular techniques could differentiate and identify Dirofilaria spp.
2. Soulsby EJL. Helminths. In: Helminths, Arthropods and Protozoa of Domesticated Animals 7th ed. Baltimore, MD: Williams & Wilkins; 1982:311.
3. Hoch H, Strickland K. Canine and feline dirofilariasis: life cycle, pathophysiology, and diagnosis. Compendium (Yardley, PA) 2008; 30: 133-40; quiz 141.
4. Canestri Trotti G, Pampiglione S, Rivasi F. The species of the genus Dirofilaria, Railliet & Henry, 1911. Parasitologia. 1997; 39: 369-74.
5. Azari-Hamidian S, Yaghoobi-Ershadi MR. Review of Dirofilariasis in Iran. J Guilan Univ Med Sci. 2007; 15: 102-14.
6. Heidari Z, Kia EB, Arzamani K et al. Morphological and molecular identification of Dirofilaria immitis from Jackal (Canis aureus) in North Khorasan, northeast Iran. J Vector Borne Dis. 2015; 52: 329-33.
7. Khedri J, Radfar MH, Borji H et al. Canine heartworm in southeastern of Iran with review of disease distribution. Iran J parasitol. 2014; 9: 560-67.
8. Alborzi A, Mosallanejad B, Najafabadi MG et al. Infestation of heartworm (Dirofilaria immitis) in a cat in Ahvaz City: a case report. J Vet Res. 2010; 65: 255-57.
9. Khodabakhsh M, Malmasi A, Mohebali M et al. Feline dirofilariosis due to Dirofilaria immitis in Meshkin Shahr district, Northwestern Iran. Iran J parasitol. 2016; 11: 269-73.
10. Rokni MB. The present status of human helminthic diseases in Iran. Ann Trop Med Parasitol. 2008; 102: 283-95.
11. Ionica AM, Matei IA, D'Amico G et al. Dirofilaria immitis and D. repens show circadian co-periodicity in naturally co-infected dogs. Parasit Vectors. 2017; 10: 116.
12. Borthakur SK, Deka DK, Islam S et al. Prevalence and molecular epidemiological data on Dirofilaria immitis in dogs from Northeastern States of India. Sci World J. 2015; 2015.
13. Albonico F, Loiacono M, Gioia G et al. Rapid differentiation of Dirofilaria immitis and Dirofilaria repens in canine peripheral blood by real-time PCR coupled to high resolution melting analysis. Vet Parasitol. 2014; 200: 128-32.
14. Zolfaghari Emameh R, Kuuslahti M, Nareaho A et al. Innovative molecular diagnosis of Trichinella species based on beta-carbonic anhydrase genomic sequence. Microb Biotechnol. 2016; 9: 172-79.
15. Zolfaghari Emameh R, Purmonen S, Sukura A et al. Surveillance and diagnosis of zoonotic foodborne parasites. Food Sci Nutr. 2017; 6: 3-17.
16. Aroch I, Rojas A, Slon P et al. Serological cross-reactivity of three commercial in-house immunoassays for detection of Dirofilaria immitis antigens with Spirocerca lupi in dogs with benign esophageal spirocercosis. Vet Parasitol. 2015; 211: 303-305.
17. Drake J, Gruntmeir J, Merritt H et al. False negative antigen tests in dogs infected with heartworm and placed on macrocyclic lactone preventives. Parasit Vectors. 2015; 8.
18. Stillman BA, Monn M, Liu J et al. Performance of a commercially available in-clinic ELISA for detection of antibodies against Anaplasma phagocytophilum, Anaplasma platys, Borrelia burgdorferi, Ehrlichia canis, and Ehrlichia ewingii and Dirofilaria immitis antigen in dogs. J Am Vet Med Assoc. 2014; 245: 80-86.
19. Solgi R, Sadjjadi SM, Mohebali M et al. Development of new recombinant DgK antigen for diagnosis of Dirofilaria immitis infections in dogs using ELISA technique and its comparison to molecular methods. Iran Biomed J. 2018; 22: 283-89.
20. Gutierrez Y. Diagnostic pathology of parasitic infections with clinical correlations, Oxford University Press, 2000:461-490.
21. Fischer AH, Jacobson KA, Rose J et al. Hematoxylin and eosin staining of tissue and cell sections. CSH protocols 2008; 49-86.
22. Khanmohammadi M, Halajian A, Ganji S. First scanning electron microscope observation on adult Oesophagostomum venulosum (Rudolphi, 1809)(Nematoda: Strongylida, Chabertiidae). Veterinarija ir Zootechnika. 2013; 62.
23. Song KH, Tanaka S, Hayasaki M. Scanning electron microscopic observation of ultrastructure of Dirofilaria immitis microfilaria. J Vet Med Sci. 2009; 71: 779-83.
24. Murata K, Yanai T, Agatsuma T et al. Dirofilaria immitis infection of a snow leopard (Uncia uncia) in a Japanese zoo with mitochondrial DNA analysis. J Vet Med Sci. 2003; 65: 945-47.
25. Furtado AP, Melo FT, Giese EG et al. Morphological redescription of Dirofilaria immitis. J Parasitol. 2010; 96: 499-504.
26. Sim C, Kim H, Son HY et al. Description of peritoneal cavity dirofilariosis caused by Dirofilaria immitis (Filarioidea: Onchocercidae) in a dog: a case report. Vet Med – Czech. 2013; 58: 105-8.
27. Kozek WJ. Ultrastructure of the microfilaria of Dirofilaria immitis. J Parasitol. 1971 1052-67.
28. Wong MM, Brummer M. Cuticular morphology of five species of Dirofilaria: A scanning electron microscope study. J Parasitol. 1978; 108-14.
29. Sano Y, Aoki M, Takahshi H et al. The first record of Dirofilaria immitis infection in a Humboldt penguin, Spheniscus humboldti. J Parasitol. 2005; 91: 1235-37.
30. Otranto D, Diniz DG, Dantas-Torres F et al. Human intraocular filariasis caused by Dirofilaria sp. nematode, Brazil. Emerg Infect Dis. 2011; 17: 863-66.
31. Rodrigues-Silva R, Lanfredi RM, Vicente JJ et al. On the rugose area and caudal papillae distribution in Dirofilaria immitis (Leidy, 1856) Railliet & Henry, 1911, by scanning electron microscopy. Parasitol Res. 1999; 85: 867-69.
32. Vezzani D, Moré GA, Eiras DF. Evidencias sobre una nueva especie del género Dirofilaria en perros de Neuquén, Argentina Analecta Veterinaria. 2017; 37.
33. Bourguinat C, Keller K, Prichard RK et al. Genetic polymorphism in Dirofilaria immitis. Vet Parasitol. 2011; 176: 368-73.
34. Rjeibi MR, Rouatbi M, Mabrouk M et al. Molecular Study of Dirofilaria immitis and Dirofilaria repens in Dogs from Tunisia. Transbound Emerg Dis. 2017, 1505-1509.
35. Bouchery T, Lefoulon E, Karadjian G et al. The symbiotic role of Wolbachia in Onchocercidae and its impact on filariasis. Clin Microbiol Rev. 2013;19:131-40.
36. Huang H, Wang T, Yang G et al. Molecular characterization and phylogenetic analysis of Dirofilaria immitis of China based on COI and 12S rDNA genes. Vet Parasitol. 2009; 160: 175-9.
|Issue||Vol 15 No 1 (2020)|
|Dirofilaria immitis Scanning electron mi-croscopy (SEM) Homology Iran|
|Rights and permissions|
|This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.|