Molecular Identification and Genotyping of Babesia canis in Dogs from Meshkin Shahr County, Northwestern Iran
,
Abstract
Background: Canine babesiosis is one of the mainly worldwide-distributed tick-borne haemoprotozoan parasitic diseases in dogs.
Methods: A total of 43 blood samples were randomly collected from naturally infected dogs in seven villages from different geographical areas of Meshkin Shahr, Ardabil Province, Iran. The presence of Babesia species detected with standard methods including parasitological and gene sequencing techniques targeting the 18S rRNA gene.
Results: Our results revealed that four dogs 9.3% (4/43) including one female and three male dogs were infected with Babesia. All four Babesia-infected dogs were confirmed B. canis by the molecular-based method. Sequence alignments comparison of the B. canis genotypes A and B, it was revealed that all B. canis isolates belonged to genotype B.
Conclusion: This study provides essential data for subsequently define the critical importance of the molecular studies in management and prevention of the canine babesiosis in Iran.
1. Solano-Gallego L, Sainz A, Roura X, Estra-da-Pena A, Miro G (2016) A review of canine babesiosis: the European per¬spec-tive. Parasit Vectors. 9(1): 336.
2. Uilenberg G (2006) Babesia a historical over¬view. Vet Parasitol. 138(1–2): 3–10.
3. Rene-Martellet M, Moro CV, Chene J, Bourdoiseau G, Chabanne L, Mavingui P (2015) Update on epidemiology of canine babesiosis in southern France. BMC Vet Res. 11: 223.
4. Wang J, Liu J, Yang J, Liu Z, Wang X, Li Y, Luo J, Guan G, Yin H (2019) Molec-ular detection and genetic diversity of Babesia canis canis in pet dogs in He-nan Province, China. Parasitol Int. 71: 37–40.
5. Zygner W, Gorski P, Wedrychowicz H (2016) New localities of Dermacentor reticula¬tus tick (vector of Babesia canis canis) in central and eastern Poland. Pol J Vet Sci. 12(4): 549–555.
6. Vichova B, Miterpakova M, Iglodyova A (2014) Molecular detection of co-infec-tions with Anaplasma phagocytophilum and/or Babesia canis canis in Dirofi-laria-positive dogs from Slovakia. Vet Parasi¬tol. 203(1–2): 167–172.
7. Rubel F, Brugger K, Pfeffer M, Chitimia-Dobler L, Didyk YM, Leverenz S, Hans D, Olaf K (2016) Geographical distribu-tion of Dermacentor marginatus and Der¬macentor reticulatus in Europe. Ticks Tick Borne Dis. 7(1): 224–233.
8. Rar VA, Maksimova TG, Zakharenko LP, Bolykhina SA, Dobrotvorsky AK, Mo-rozova OV (2005) Babesia DNA detec-tion in canine blood and Dermacentor reticulatus ticks in southwestern Sibe¬ria, Russia. Vector Borne Zoonotic Dis. 5(3): 285–287.
9. Orkun O, Karaer Z (2017) Molecular char-acterization of Babesia species in wild animals and their ticks in Turkey. Infect Genet Evol Infect. 55: 8–13.
10. Aktas M, Ozubek S, Altay K, Ipek ND, Balkaya I, Utuk AE, Erdem Utuk A, Kır¬bas A, Şimsek S, Dumanlı N (2015) Mo¬lecular detection of tick-borne rickettsial and protozoan pathogens in domestic dogs from Turkey. Parasit Vectors. 8: 157.
11. Mazloum Z (1971) Ticks of domestic ani-mals in Iran: Geographical distribution, host relation and seasonal activity. Iran J Vet Med. 27: 1–32.
12. Gholamreza S, Somaieh M, Roya S, Ali-reza B, Ghazale A, Yasin B (2017) First detection of Babesia ovis in Dermacen-tor spp in Ardabil area, northwest of Iran. J Vector Borne Dis. 54(3): 277–81.
13. Esmaeilnejad B TM, Asri-Rezaei S, Dalir-Naghadeh B, Mardani K, Golabi M, Arjmand J, Kazemnia A, Jalilzadeh G (2015) Determination of prevalence and risk factors of infection with Babesia ovis in small ruminants from West Azerbai¬jan Province, Iran by polymerase chain reaction. Iran J Arthropod Borne Dis. 9 (2): 246–252.
14. Nabian S, Rahbari S, Shayan P, Hadadza-deh HR (2008) Identification of tick spe¬cies of dermacentor in some localities of Iran. J Vet Res. 63(3): 123–126.
15. Rahbari S, Nabian S, Shayan P (2007) Pri-mary report on distribution of tick fauna in Iran. Parasitol Res. 101 Suppl 2: 175–177.
16. Razmi GR, Glinsharifodini M, Sarvi S (2007) Prevalence of ixodid ticks on cat¬tle in Mazandaran Province, Iran. Kore¬an J Parasitol. 45(4): 307–310.
17. Nabian S, Shayan P, Haddadzadeh HR (2007) Current status of tick fauna in north of Iran. Iran J Parasitol. 2(1): 12–17.
18. Akhtardanesh B, Saberi M, Nurollahifard SR, Aghazamani M (2016) Molecular de¬tection of Babesia spp. in tick-infested dogs in southeastern Iran. J Dis Glob Health. 8(2): 72–77.
19. Bigdeli M, Rafie SM, Namavari MM, Jam¬shidi S (2012) Report of Theileria annu¬lata and Babesia canis infections in dogs. Comp Clin Path. 21(3): 375–377.
20. Jalali R, Mosallanejad MH, Avizeh B, Al-borzi R, Hamidi Nejat AR, Taghipour H (2013) Babesia infection in urban and ru¬ral dogs in Ahvaz District, Southwest of Iran. Arch Razi Inst. 68(1): 37–42.
21. Niak A, Anwar M, Khatibi S (1973) Ca-nine babesiosis in Iran. Trop Anim Health Prod. 5(3): 200–201.
22. Augustine S, Sabu L, Lakshmanan B (2017) Molecular identification of Babesia spp. in naturally infected dogs of Kerala, South India. J Parasit Dis. 41(2): 459–462.
23. Bajer A, Rodo A, Bednarska M, Mierzejew¬ska E, Welc-Faleciak R (2013) Babesia canis and tick-borne encephalitis virus (TBEV) co-infection in a sled dog. Ann Agric Environ Med. 20(3): 426–430.
24. Beck A, Huber D, Polkinghorne A, Kurilj AG, Benko V, Mrljak V (2017) The prev¬alence and impact of Babesia canis and Theileria sp. in free-ranging grey wolf (Canis lupus) populations in Croatia. Par¬asite Vectors. 10(1): 168.
25. Ionita M, Mitrea IL, Pfister K, Hamel D, Buzatu CM, Silaghi C (2012) Canine babesiosis in Romania due to Babesia canis and Babesia vogeli: a molecular ap¬proach. Parasitol Res. 110(5): 1659–1664.
26. Juwaid S, Sukara R, Penezic A, Mihaljica D, Veinovic G, Kavallieratos NG, Ćirović D, Tomanović S (2019) First evidence of tick-borne protozoan pathogens, Babesia sp. and Hepatozoon canis, in red foxes (vulpes vulpes) in Serbia. Acta Vet Hung. 67(1): 70–80.
27. Sukara R, Chochlakis D, Cirovic D, Pene-zic A, Mihaljica D, Cakic S, Valčić M, Tselentis Y, Psaroulaki A, Tomanović S (2018) Golden jackals (Canis aureus) as hosts for ticks and tick-borne pathogens in Serbia. Ticks Tick Borne Dis. 9(5): 1090–1097.
28. Solano-Gallego L, Baneth G (2011) Babesi¬osis in dogs and cats-expanding para¬si¬tological and clinical spectra. Vet par¬a¬sitol. 181(1): 48–60.
29. Baneth G, Kenny MJ, Tasker S, Anug Y, Shkap V, Levy A, Shaw SE (2004) In-fection with a proposed new subspecies of Babesia canis, Babesia canis subsp. presentii, in domestic cats. J Clin Mi¬cro-biol. 42(1): 99–105.
30. Brkljacic M, Matijatko V, Kis I, Kucer N, Forsek J, Rafaj RB, Grden D, Torti M, Mayer I, Mrljak V (2010) Molecular ev-idence of natural infection with Babesia canis canis in Croatia. Acta Vet Hung. 58(1): 39–46.
31. Lyp P, Adaszek L, Furmaga B, Winiar-czyk S (2015) Identification of new 18S rRNA strains of Babesia canis isolated from dogs with subclinical babesiosis. Pol J Vet. 18(3): 573–577.
32. Sanchez-Villeda H, Schroeder S, Flint-Gar¬cia S, Guill KE, Yamasaki M, McMul¬len MD (2008) DNA Align Editor: DNA alignment editor tool. BMC Bioinfor¬mat¬ics. 9: 154.
33. Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sen¬sitivity of progressive multiple sequence alignment through sequence weighting, po¬sition-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22 (22): 4673–4680.
34. Nishimaki T, Sato K (2019) An Extension of the Kimura Two-Parameter Model to the Natural Evolutionary Process. J Mol Evol. 87(1): 60–67.
35. Adaszek L, Winiarczyk S (2008) Molecu-lar characterization of Babesia canis canis isolates from naturally infected dogs in Poland. Vet Parasitol. 152(3–4): 235–241.
36. Hrazdilova K, Mysliwy I, Hildebrand J, Bun¬kowska-Gawlik K, Janaczyk B, Perec-Matysiak A, Modrýa D (2019) Paralogs vs. genotypes, variability of Babesia canis assessed by 18S rDNA and two mitochondrial markers. Vet Parasitol. 266: 103–110.
37. Ashrafi H, Haddadzadeh H, Shirani D, Khazrai¬inia P, Mostofi S (2001) Histo-pathologic, hematologic and clinical study on canine babesiosis. Iran J Vet Med. 56: 93–96.
38. Annoscia G, Latrofa MS, Cantacessi C, Olivieri E, Manfredi MT, Dantas-Torres F, Otranto D (2017) A new PCR assay for the detection and differentiation of Babesia canis and Babesia vogeli. Ticks Tick Borne Dis. 8(6): 862–865.
39. Milanović Z, Vekić J, Radonjić V, Ilić Božović A, Zeljković A, Janac J, Spaso-jević-Kalimanovska V, Buch J, Chan-drashekar R, Bojić-Trbojević Ž, Hajdu-ković L, Christopher MM, Kovačević Fil¬ipović M (2019) Association of acute Babesia canis infection and serum lipid, lipoprotein, and apoprotein concentra-tions in dogs. J Vet Intern Med. 33(4): 1686–1694.
40. Davitkov D, Vucicevic M, Stevanovic J, Krstic V, Tomanovic S, Glavinic U, Stanimi¬rovic Z (2015) Clinical babesio-sis and molecular identification of Babe-sia canis and Babesia gibsoni infections in dogs from Serbia. Acta Vet Hung. 63 (2): 199–208.
41. Kovacevic Filipovic MM, Beletic AD, Ilic Bozovic AV, Milanovic Z, Tyrrell P, Buch J, Breitschwerdt EB, Birkenheuer AJ, Chandrashekar R (2018) Molecular and serological prevalence of Anaplas-ma phagocytophilum, A. platys, Ehr-lichia canis, E. chaffeenses, E. ewingii, Borre¬lia burgdorferi, Babesia canis, B. gib¬soni and B. vogeli among clinically healthy outdoor dogs in Serbia. Vet Parasitol Reg Stud Reports. 14: 117–122.
42. Beugnet F, Chalvet-Monfray K (2013) Im¬pact of climate change in the epidemiol¬ogy of vector-borne diseases in domes¬tic carnivores. Comp Immunol Microbi¬ol Infect Dis. 36(6): 559–566.
2. Uilenberg G (2006) Babesia a historical over¬view. Vet Parasitol. 138(1–2): 3–10.
3. Rene-Martellet M, Moro CV, Chene J, Bourdoiseau G, Chabanne L, Mavingui P (2015) Update on epidemiology of canine babesiosis in southern France. BMC Vet Res. 11: 223.
4. Wang J, Liu J, Yang J, Liu Z, Wang X, Li Y, Luo J, Guan G, Yin H (2019) Molec-ular detection and genetic diversity of Babesia canis canis in pet dogs in He-nan Province, China. Parasitol Int. 71: 37–40.
5. Zygner W, Gorski P, Wedrychowicz H (2016) New localities of Dermacentor reticula¬tus tick (vector of Babesia canis canis) in central and eastern Poland. Pol J Vet Sci. 12(4): 549–555.
6. Vichova B, Miterpakova M, Iglodyova A (2014) Molecular detection of co-infec-tions with Anaplasma phagocytophilum and/or Babesia canis canis in Dirofi-laria-positive dogs from Slovakia. Vet Parasi¬tol. 203(1–2): 167–172.
7. Rubel F, Brugger K, Pfeffer M, Chitimia-Dobler L, Didyk YM, Leverenz S, Hans D, Olaf K (2016) Geographical distribu-tion of Dermacentor marginatus and Der¬macentor reticulatus in Europe. Ticks Tick Borne Dis. 7(1): 224–233.
8. Rar VA, Maksimova TG, Zakharenko LP, Bolykhina SA, Dobrotvorsky AK, Mo-rozova OV (2005) Babesia DNA detec-tion in canine blood and Dermacentor reticulatus ticks in southwestern Sibe¬ria, Russia. Vector Borne Zoonotic Dis. 5(3): 285–287.
9. Orkun O, Karaer Z (2017) Molecular char-acterization of Babesia species in wild animals and their ticks in Turkey. Infect Genet Evol Infect. 55: 8–13.
10. Aktas M, Ozubek S, Altay K, Ipek ND, Balkaya I, Utuk AE, Erdem Utuk A, Kır¬bas A, Şimsek S, Dumanlı N (2015) Mo¬lecular detection of tick-borne rickettsial and protozoan pathogens in domestic dogs from Turkey. Parasit Vectors. 8: 157.
11. Mazloum Z (1971) Ticks of domestic ani-mals in Iran: Geographical distribution, host relation and seasonal activity. Iran J Vet Med. 27: 1–32.
12. Gholamreza S, Somaieh M, Roya S, Ali-reza B, Ghazale A, Yasin B (2017) First detection of Babesia ovis in Dermacen-tor spp in Ardabil area, northwest of Iran. J Vector Borne Dis. 54(3): 277–81.
13. Esmaeilnejad B TM, Asri-Rezaei S, Dalir-Naghadeh B, Mardani K, Golabi M, Arjmand J, Kazemnia A, Jalilzadeh G (2015) Determination of prevalence and risk factors of infection with Babesia ovis in small ruminants from West Azerbai¬jan Province, Iran by polymerase chain reaction. Iran J Arthropod Borne Dis. 9 (2): 246–252.
14. Nabian S, Rahbari S, Shayan P, Hadadza-deh HR (2008) Identification of tick spe¬cies of dermacentor in some localities of Iran. J Vet Res. 63(3): 123–126.
15. Rahbari S, Nabian S, Shayan P (2007) Pri-mary report on distribution of tick fauna in Iran. Parasitol Res. 101 Suppl 2: 175–177.
16. Razmi GR, Glinsharifodini M, Sarvi S (2007) Prevalence of ixodid ticks on cat¬tle in Mazandaran Province, Iran. Kore¬an J Parasitol. 45(4): 307–310.
17. Nabian S, Shayan P, Haddadzadeh HR (2007) Current status of tick fauna in north of Iran. Iran J Parasitol. 2(1): 12–17.
18. Akhtardanesh B, Saberi M, Nurollahifard SR, Aghazamani M (2016) Molecular de¬tection of Babesia spp. in tick-infested dogs in southeastern Iran. J Dis Glob Health. 8(2): 72–77.
19. Bigdeli M, Rafie SM, Namavari MM, Jam¬shidi S (2012) Report of Theileria annu¬lata and Babesia canis infections in dogs. Comp Clin Path. 21(3): 375–377.
20. Jalali R, Mosallanejad MH, Avizeh B, Al-borzi R, Hamidi Nejat AR, Taghipour H (2013) Babesia infection in urban and ru¬ral dogs in Ahvaz District, Southwest of Iran. Arch Razi Inst. 68(1): 37–42.
21. Niak A, Anwar M, Khatibi S (1973) Ca-nine babesiosis in Iran. Trop Anim Health Prod. 5(3): 200–201.
22. Augustine S, Sabu L, Lakshmanan B (2017) Molecular identification of Babesia spp. in naturally infected dogs of Kerala, South India. J Parasit Dis. 41(2): 459–462.
23. Bajer A, Rodo A, Bednarska M, Mierzejew¬ska E, Welc-Faleciak R (2013) Babesia canis and tick-borne encephalitis virus (TBEV) co-infection in a sled dog. Ann Agric Environ Med. 20(3): 426–430.
24. Beck A, Huber D, Polkinghorne A, Kurilj AG, Benko V, Mrljak V (2017) The prev¬alence and impact of Babesia canis and Theileria sp. in free-ranging grey wolf (Canis lupus) populations in Croatia. Par¬asite Vectors. 10(1): 168.
25. Ionita M, Mitrea IL, Pfister K, Hamel D, Buzatu CM, Silaghi C (2012) Canine babesiosis in Romania due to Babesia canis and Babesia vogeli: a molecular ap¬proach. Parasitol Res. 110(5): 1659–1664.
26. Juwaid S, Sukara R, Penezic A, Mihaljica D, Veinovic G, Kavallieratos NG, Ćirović D, Tomanović S (2019) First evidence of tick-borne protozoan pathogens, Babesia sp. and Hepatozoon canis, in red foxes (vulpes vulpes) in Serbia. Acta Vet Hung. 67(1): 70–80.
27. Sukara R, Chochlakis D, Cirovic D, Pene-zic A, Mihaljica D, Cakic S, Valčić M, Tselentis Y, Psaroulaki A, Tomanović S (2018) Golden jackals (Canis aureus) as hosts for ticks and tick-borne pathogens in Serbia. Ticks Tick Borne Dis. 9(5): 1090–1097.
28. Solano-Gallego L, Baneth G (2011) Babesi¬osis in dogs and cats-expanding para¬si¬tological and clinical spectra. Vet par¬a¬sitol. 181(1): 48–60.
29. Baneth G, Kenny MJ, Tasker S, Anug Y, Shkap V, Levy A, Shaw SE (2004) In-fection with a proposed new subspecies of Babesia canis, Babesia canis subsp. presentii, in domestic cats. J Clin Mi¬cro-biol. 42(1): 99–105.
30. Brkljacic M, Matijatko V, Kis I, Kucer N, Forsek J, Rafaj RB, Grden D, Torti M, Mayer I, Mrljak V (2010) Molecular ev-idence of natural infection with Babesia canis canis in Croatia. Acta Vet Hung. 58(1): 39–46.
31. Lyp P, Adaszek L, Furmaga B, Winiar-czyk S (2015) Identification of new 18S rRNA strains of Babesia canis isolated from dogs with subclinical babesiosis. Pol J Vet. 18(3): 573–577.
32. Sanchez-Villeda H, Schroeder S, Flint-Gar¬cia S, Guill KE, Yamasaki M, McMul¬len MD (2008) DNA Align Editor: DNA alignment editor tool. BMC Bioinfor¬mat¬ics. 9: 154.
33. Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sen¬sitivity of progressive multiple sequence alignment through sequence weighting, po¬sition-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22 (22): 4673–4680.
34. Nishimaki T, Sato K (2019) An Extension of the Kimura Two-Parameter Model to the Natural Evolutionary Process. J Mol Evol. 87(1): 60–67.
35. Adaszek L, Winiarczyk S (2008) Molecu-lar characterization of Babesia canis canis isolates from naturally infected dogs in Poland. Vet Parasitol. 152(3–4): 235–241.
36. Hrazdilova K, Mysliwy I, Hildebrand J, Bun¬kowska-Gawlik K, Janaczyk B, Perec-Matysiak A, Modrýa D (2019) Paralogs vs. genotypes, variability of Babesia canis assessed by 18S rDNA and two mitochondrial markers. Vet Parasitol. 266: 103–110.
37. Ashrafi H, Haddadzadeh H, Shirani D, Khazrai¬inia P, Mostofi S (2001) Histo-pathologic, hematologic and clinical study on canine babesiosis. Iran J Vet Med. 56: 93–96.
38. Annoscia G, Latrofa MS, Cantacessi C, Olivieri E, Manfredi MT, Dantas-Torres F, Otranto D (2017) A new PCR assay for the detection and differentiation of Babesia canis and Babesia vogeli. Ticks Tick Borne Dis. 8(6): 862–865.
39. Milanović Z, Vekić J, Radonjić V, Ilić Božović A, Zeljković A, Janac J, Spaso-jević-Kalimanovska V, Buch J, Chan-drashekar R, Bojić-Trbojević Ž, Hajdu-ković L, Christopher MM, Kovačević Fil¬ipović M (2019) Association of acute Babesia canis infection and serum lipid, lipoprotein, and apoprotein concentra-tions in dogs. J Vet Intern Med. 33(4): 1686–1694.
40. Davitkov D, Vucicevic M, Stevanovic J, Krstic V, Tomanovic S, Glavinic U, Stanimi¬rovic Z (2015) Clinical babesio-sis and molecular identification of Babe-sia canis and Babesia gibsoni infections in dogs from Serbia. Acta Vet Hung. 63 (2): 199–208.
41. Kovacevic Filipovic MM, Beletic AD, Ilic Bozovic AV, Milanovic Z, Tyrrell P, Buch J, Breitschwerdt EB, Birkenheuer AJ, Chandrashekar R (2018) Molecular and serological prevalence of Anaplas-ma phagocytophilum, A. platys, Ehr-lichia canis, E. chaffeenses, E. ewingii, Borre¬lia burgdorferi, Babesia canis, B. gib¬soni and B. vogeli among clinically healthy outdoor dogs in Serbia. Vet Parasitol Reg Stud Reports. 14: 117–122.
42. Beugnet F, Chalvet-Monfray K (2013) Im¬pact of climate change in the epidemiol¬ogy of vector-borne diseases in domes¬tic carnivores. Comp Immunol Microbi¬ol Infect Dis. 36(6): 559–566.
| Files | ||
| Issue | Vol 15 No 1 (2021) | |
| Section | Original Article | |
| DOI | https://doi.org/10.18502/jad.v15i1.6489 | |
| Keywords | ||
| Babesia canis; Babesiosis; Dogs; Genotyping; RNA, ribosomal, 18S; Iran | ||
| Rights and permissions | |
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This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Khanmohammadi M, Zolfaghari-Emameh R, Arshadi M, Razmjou E, Karimi P. Molecular Identification and Genotyping of Babesia canis in Dogs from Meshkin Shahr County, Northwestern Iran. J Arthropod Borne Dis. 2021;15(1):97-107.
