Evaluation of prevalence and phylogenetic analysis of Wolbachia pipientis isolated from Dirofilaria immitis of canids in northern Iran

Meysam  Sharifdini; Hadi  Sedigh Ebrahim-Saraie; Meysam  Hasannejad-Bibalan; Ali  Mirsaeidi; Edris  Mehmandoost Du; Farid  Ajori

doi:10.61882/jcbior.6.1.309

Authors

Meysam Sharifdini Department of Medical Parasitology and Mycology, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran https://orcid.org/0000-0003-2686-7748
Hadi Sedigh Ebrahim-Saraie Department of Microbiology, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran
Meysam Hasannejad-Bibalan Department of Microbiology, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran
Ali Mirsaeidi Student Research Committee, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran
Edris Mehmandoost Du Department of Microbiology, Faculty of Medicine, Shahed University, Tehran, Iran
Farid Ajori Student Research Committee, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran

DOI:

https://doi.org/10.61882/jcbior.6.1.309

Keywords:

Dirofilaria, Nematodes, Wolbachia, Phylogenetic, Carnivores

Abstract

Dirofilaria immitis and Dirofilaria repens, two mosquito-borne filarial nematodes, are the primary causes of subcutaneous and cardiopulmonary dirofilariasis in humans and canines. Wolbachia pipientis infection is widespread among arthropods and nematodes globally. One safe, cost-effective, and efficient way to avoid dirofilariasis infections is to eradicate Wolbachia. So, herein we determined the prevalence and phylogenetic analysis of W. pipientis strains isolated from D. immitis of canids in northern Iran. A descriptive-analytical cross-sectional study from March 2019 to April 2020 in Guilan province, north of Iran, was conducted. Sampling consisted of 32 road-killed carnivores (12 Canis familiaris and 20 Canis aureus) were gathered and necropsied for this study. The 16S ribosomal RNA (rRNA) gene was the basis for the molecular analysis. D. immitis infection was discovered in 7/20 (35%) of the jackals and 9/12 (75%) of the dogs. The W. pipientis infection was present in all 16 of the infected dogs and jackals. In this study, 16S rRNA sequences had 100% similarity with previously submitted W. pipientis sequences from the USA, Russia, Myanmar, and Italy. This paper hopes to provide a new vision for further studies on the symbiotic relationship between D. immitis and Wolbachia, providing an advance in the therapeutic and diagnostic approach. There is no report of molecular identification for Wolbachia spp. isolated from D. immitis in northern Iran. So, to fill this study gap, herein we determined the prevalence and phylogenetic analysis of W. pipientis strains isolated from D. immitis of canids in northern Iran.

Author Biography

Hadi Sedigh Ebrahim-Saraie, Department of Microbiology, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran

References

1. Shirozu T, Regilme MAF, Ote M, Sasaki M, Soga A, Bochimoto H, et al. Wolbachia infection in Aedes aegypti does not affect its vectorial capacity for Dirofilaria immitis. Sci Rep. 2024;14(1):22528.

DOI: 10.1038/s41598-024-73421-9 PMID: 39341970

2. Orihel TC, Eberhard ML. Zoonotic filariasis. Clin Microbiol Rev. 1998;11(2):366-81.

DOI: 10.1128/CMR.11.2.366 PMID: 9564568

3. Carvajal TM, Hashimoto K, Harnandika RK, Amalin DM, Watanabe K. Detection of Wolbachia in field-collected Aedes aegypti mosquitoes in metropolitan Manila, Philippines. Parasit Vectors. 2019;12(1):361. DOI: 10.1186/s13071-019-3629-y PMID: 31340862

4. Sarwar MS, Jahan N, Shahbaz F. Molecular Detection and Characterization of Wolbachia pipientis from Culex quinquefasciatus Collected from Lahore, Pakistan. Am J Trop Med Hyg. 2018;98(1):154-161. DOI: 10.4269/ajtmh.17-0329 PMID: 29141766

5. Younes L, Barré-Cardi H, Bedjaoui S, Ayhan N, Varloud M, Mediannikov O, et al. Dirofilaria immitis and Dirofilaria repens in mosquitoes from Corsica Island, France. Parasit Vectors. 2021;14(1):427. DOI: 10.1186/s13071-021-04931-y

PMID: 34446069

6. Kaiser C, Müller P, Zinsstag J. Occurrence of canine subcutaneous dirofilariosis (Dirofilaria repens) in Switzerland [German]. Schweiz Arch Tierheilkd. 2023;165(5):321-31. URL: https://sat.gstsvs.ch/fileadmin/datapool_upload/IgJournal/Artikel/pdf/321_331_Kaiser_02.pdf

7. Epis S, Varotto-Boccazzi I, Crotti E, Damiani C, Giovati L, Mandrioli M, et al. Chimeric symbionts expressing a Wolbachia protein stimulate mosquito immunity and inhibit filarial parasite development. Commun Biol. 2020;3(1):105.

DOI: 10.1038/s42003-020-0835-2 PMID: 32144396

8. Bawm S, Khaing Y, Chel HM, Hmoon MM, Win SY, Bo M, et al. Molecular detection of Dirofilaria immitis and its Wolbachia endosymbionts in dogs from Myanmar. Curr Res Parasitol Vector Borne Dis. 2023;4:100148. DOI: 10.1016/j.crpvbd.2023.100148 PMID: 38021190

9. de Lavalle Galvis R, de La Puente M, Socarras TdJO, et al. Wolbachia pipientis: Endosymbiont Bacteria of Dirofilaria immitis and its Role in Feline Dirofilariasis. Aquac Aquar Conserv Legis. 2020;12(2):59-64.

URL: http://www.hvm.bioflux.com.ro/docs/2020.59-64a.pdf

10. Kang Y, Dempsey B. Investigating the presence of Wolbachia pipientis in various mosquito species. J Exp Second Sci. 2011;1:1-4.

URL: https://www.researchgate.net/publication/216466570

11. Kamkong P, Jitsamai W, Thongmeesee K, Ratthawongjirakul P, Taweethavonsawat P. Genetic diversity and characterization of Wolbachia endosymbiont in canine filariasis. Acta Trop. 2023;246:107000. DOI: 10.1016/j.actatropica.2023.107000 PMID: 37567493

12. Khanmohammadi M, Falak R, Meamar AR, Arshadi M, Akhlaghi L, Razmjou E. Molecular Detection and Phylogenetic Analysis of Endosymbiont Wolbachia pipientis (Rickettsiales: Anaplasmataceae) Isolated from Dirofilaria immitis in Northwest of Iran. J Arthropod Borne Dis. 2019;13(1):83-93.

PMID: 31346538

13. Louzada-Flores VN, Kramer L, Brianti E, Napoli E, Mendoza-Roldan JA, Bezerra-Santos MA, et al. Treatment with doxycycline is associated with complete clearance of circulating Wolbachia DNA in Dirofilaria immitis-naturally infected dogs. Acta Trop. 2022;232:106513.

DOI: 10.1016/j.actatropica.2022.106513 PMID: 35598650

14. Sharifdini M, Karimi M, Ashrafi K, Soleimani M, Mirjalali H. Prevalence and molecular characterization of Dirofilaria immitis in road killed canids of northern Iran. BMC Vet Res. 2022;18(1):161. DOI: 10.1186/s12917-022-03270-z

PMID: 35501899

15. Bereczki J, Rácz R, Varga Z, Tóth JP. Controversial patterns of Wolbachia infestation in the social parasitic Maculinea butterflies (Lepidoptera: Lycaenidae). Organisms Diversity & Evolution. 2015;15(3):591-607. DOI: 10.1007/s13127-015-0217-7

16. DE Pinho Mixão V, Mendes AM, Maurício IL, Calado MM, Novo MT, Belo S, et al. Molecular detection of Wolbachia pipientis in natural populations of mosquito vectors of Dirofilaria immitis from continental Portugal: first detection in Culex theileri. Med Vet Entomol. 2016;30(3):301-9.

DOI: 10.1111/mve.12179 PMID: 27279553

17. Dyab AK, Galal LA, Mahmoud AE, Mokhtar Y. Finding Wolbachia in Filarial larvae and Culicidae Mosquitoes in Upper Egypt Governorate. Korean J Parasitol. 2016;54(3):265-72.

DOI: 10.3347/kjp.2016.54.3.265 PMID: 27417080

18. Karimi J, Darsouei R. Presence of the endosymbiont Wolbachia among some fruit flies (Diptera: Tephritidae) from Iran: A multilocus sequence typing approach. Journal of Asia-Pacific Entomology. 2014;17(1):105-12.

DOI: 10.1016/j.aspen.2013.11.002

19. Parvizi P, Fardid F, Soleimani S. Detection of a New Strain of Wolbachia pipientis in Phlebotomus perfiliewi transcaucasicus, a Potential Vector of Visceral Leishmaniasis in North West of Iran, by Targeting the Major Surface Protein Gene. J Arthropod Borne Dis. 2013;7(1):46-55. PMID: 23785694

20. Poorjavad N, Goldansaz SH, Machtelinckx T, Tirry L, Stouthamer R, Van Leeuwen T. Iranian Trichogramma: ITS2 DNA characterization and natural Wolbachia infection. BioControl. 2012;57(3):361-74.

DOI: 10.1007/s10526-011-9397-z

21. Teixeira L, Ferreira A, Ashburner M. The bacterial symbiont Wolbachia induces resistance to RNA viral infections in Drosophila melanogaster. PLoS Biol. 2008;6(12):e2.

DOI: 10.1371/journal.pbio.1000002 PMID: 19222304

22. Tijsse-Klasen E, Braks M, Scholte EJ, Sprong H. Parasites of vectors--Ixodiphagus hookeri and its Wolbachia symbionts in ticks in The Netherlands. Parasit Vectors. 2011;4:228.

DOI: 10.1186/1756-3305-4-228 PMID: 22152674

23. Zha X, Zhang W, Zhou C, Zhang L, Xiang Z, Xia Q. Detection and characterization of Wolbachia infection in silkworm. Genet Mol Biol. 2014;37(3):573-80.

DOI: 10.1590/s1415-47572014000400014 PMID: 25249781

24. Bandi C, Trees AJ, Brattig NW. Wolbachia in filarial nematodes: evolutionary aspects and implications for the pathogenesis and treatment of filarial diseases. Vet Parasitol. 2001;98(1-3):215-38. DOI: 10.1016/s0304-4017(01)00432-0 PMID: 11516587

25. Rossi MI, Aguiar-Alves F, Santos S, Paiva J, Bendas A, Fernandes O, et al. Detection of Wolbachia DNA in blood from dogs infected with Dirofilaria immitis. Exp Parasitol. 2010;126(2):270-2. DOI: 10.1016/j.exppara.2010.05.002

PMID: 20457156

26. Tabar MD, Altet L, Martínez V, Roura X. Wolbachia, filariae and Leishmania coinfection in dogs from a Mediterranean area. J Small Anim Pract. 2013;54(4):174-8. DOI: 10.1111/jsap.12041 PMID: 23425244

27. Comandatore F, Cordaux R, Bandi C, Blaxter M, Darby A, Makepeace BL, et al. Supergroup C Wolbachia, mutualist symbionts of filarial nematodes, have a distinct genome structure. Open Biol. 2015;5(12):150099. DOI: 10.1098/rsob.150099 PMID: 26631376

28. Enemark HL, Oksanen A, Chriél M, le Fèvre Harslund J, Woolsey ID, Al-Sabi MN. Detection and molecular characterization of the mosquito-borne filarial nematode Setaria tundra in Danish roe deer (Capreolus capreolus). Int J Parasitol Parasites Wildl. 2017;6(1):16-21. DOI: 10.1016/j.ijppaw.2017.01.002

PMID: 28229043

29. Taylor MJ, Voronin D, Johnston KL, Ford L. Wolbachia filarial interactions. Cell Microbiol. 2013;15(4):520-6.

DOI: 10.1111/cmi.12084 PMID: 23210448

30. Hosseini SH, Manshori-Ghaishghorshagh F, Ramezani M, Nayebzadeh H, Ahoo MB, Eslamian A, et al. Canine microfilaraemia in some regions of Iran. Parasit Vectors. 2022;15(1):90. DOI: 10.1186/s13071-022-05209-7

PMID: 35303931

31. Khedri J, Radfar MH, Borji H, Azizzadeh M, Akhtardanesh B. Canine Heartworm in Southeastern of Iran with Review of disease distribution. Iran J Parasitol. 2014;9(4):560-7. PMID: 25759738

32. Ranjbar-Bahadori Sh, Veshgini A, Shirani D, Eslami A, Mohieddin H, Shemshadi B, et al. Epidemiological aspects of canine dirofilariasis in the north of iran. Iran J Parasitol. 2011;6(1):73-80. PMID: 22347277