Open Access
Research (Published online: 28-09-2018)
22. Detection of Toxoplasma gondii copro-prevalence by polymerase chain reaction using repetitive 529 bp gene in feces of pet cats (Felis catus) in Yogyakarta, Indonesia
Muhammad Hanafiah, Joko Prastowo, Sri Hartati, Dwinna Aliza and Raden Wisnu Nurcahyo
Veterinary World, 11(9): 1338-1343

Muhammad Hanafiah: Parasitology Laboratory, Faculty of Veterinary Medicine, Syiah Kuala University, Banda Aceh, Indonesia.
Joko Prastowo: Department of Parasitology, Faculty of Veterinary Medicine, Gadjah Mada University, Yogyakarta, Indonesia.
Sri Hartati: Department of Clinic, Faculty of Veterinary Medicine, Gadjah Mada University, Yogyakarta, Indonesia.
Dwinna Aliza: Pathology Laboratory, Faculty of Veterinary Medicine, Syiah Kuala University, Banda Aceh, Indonesia.
Raden Wisnu Nurcahyo: Department of Parasitology, Faculty of Veterinary Medicine, Gadjah Mada University, Yogyakarta, Indonesia.

doi: 10.14202/vetworld.2018.1338-1343

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Article history: Received: 26-04-2018, Accepted: 31-07-2018, Published online: 28-09-2018

Corresponding author: Raden Wisnu Nurcahyo


Citation: Hanafiah M, Prastowo J, Hartati S, Aliza D, Nurcahyo RW (2018) Detection of Toxoplasma gondii copro-prevalence by polymerase chain reaction using repetitive 529 bp gene in feces of pet cats (Felis catus) in Yogyakarta, Indonesia, Veterinary World, 11(9): 1338-1343.

Aim: The aim of this research was to determine the copro-prevalence of Toxoplasma gondii using polymerase chain reaction (PCR) with repetitive 529 bp gene and to construct the phylogenetic tree of Toxoplasma oocyst from pet cats in Yogyakarta.

Materials and Methods: 9 of 132 pet cat samples which serologically positive for Toxoplasma were used in this research. To determine the copro-prevalence of T. gondii in pet cat, 10 g of feces samples taken from practitioners and household cats in Yogyakarta were used in the PCR method utilizing repetitive 529 bp gene sequences.

Results: The result shows that copro-prevalence by PCR using repetitive 529 bp gene was 33.3% (3/9). The phylogenetic tree of Toxoplasma grouped into two clades, which clade 1 consists of Toxoplasma isolates collected from pet cats in Yogyakarta Indonesia and T. gondii isolates from China and in clade 2 consist of the T. gondii isolates from India.

Conclusion: Copro-prevalence of T. gondii in pet cats in Yogyakarta by means of PCR using repetitive 529 bp gene is around 33.3%.

Keywords: copro-prevalence, pet cat, polymerase chain reaction, Toxoplasma gondii.


1. Hill, D.E. and Dubey, J.P. (2013) Toxoplasma gondii prevalence in farm animals in the United States. Int. J. Parasitol., 43(2): 107-113. [Crossref] [PubMed]

2. Elmore, S.A., Jones, J.L., Conrad, P.A., Patton, S., Lindsay, D.S. and Dubey, J.P. (2010) Toxoplasma gondii: Epidemiology, feline aspects, and prevention. Trends Parasitol., 26(4): 190-196. [Crossref] [PubMed]

3. Riaz, F., Rashid, M., Akbar, H., Shehzad, W., Islam, S., Arshad, B., Saeed, K. and Ashraf, K (2016) DNA amplification techniques for the detection of Toxoplasma gondii tissue cysts in meat producing animals: A narrative review article. Iran. J. Parasitol.,11(4): 431-440. [PubMed] [PMC]

4. Jones, J.L., Moran, K.D., Rivera, H.N., Price, C. and Wilkins, P.P. (2014) Toxoplasma gondii seroprevalence in the United States 2009-2010 and comparison with the past two decades. Am. J. Trop. Med. Hyg., 90(6): 1135-1139. [Crossref] [PubMed] [PMC]

5. Loss, S.R., Will, T. and Marra, P.P. (2013) The impact of free-ranging domestic cats on wildlife of the United States. Nat. Comm, 4:1396. [Crossref] [PubMed]

6. Hanafiah, M., Nurcahyo, R.W., Yuniar, R.S., Prastowo, J., Hartati, S., Sutrisno, B. and Aliza, D. (2017) Detection of Toxoplasma gondii in cat's internal organs by immunohistochemistry methods labeled with-[strept] avidin-biotin. Vet. World, 10(9): 1035-1039. [Crossref] [PubMed] [PMC]

7. Dehkordi, F.S., Haghighi, B.M.R., Rahimi, E. and Abdizadeh, R. (2013) Detection of Toxoplasma gondii in raw caprine, ovine, buffalo, bovine, and camel milk using cell cultivation, cat bioassay, capture ELISA, and PCR methods in Iran. Foodborne Pathog. Dis., 10(2): 120-125. [Crossref] [PubMed]

8. Contini, C., Seraceni, S., Cultrera, R., Incorvaia, C., Sebastiani, A. and Picot, S. (2005) Evaluation of a real-time PCR-based assay using the lightcycler system for detection of Toxoplasma gondii bradyzoite genes in blood specimens from patients with toxoplasmic retinochoroiditis. Int. J. Parasitol., 35(3): 275-283. [Crossref] [PubMed]

9. Caldearo, A., Piccolo, G., Gorrini, C., Peruzzi, S., Zerbini, L., Bommezzadri, S., Dettori, G. and Chezzi, C. (2006) Comparison between two real-time PCR assays and a nested-PCR for the detection of Toxoplasma gondii. Acta Bio.Med., 77(2): 75-80.

10. Bastien, P., Jumas-Bilak, E., Varlet-Marie, E. and Marty, P. (2007) Three years of multi-laboratory external quality control for the molecular detection of Toxoplasma gondii in amniotic fluid in France. Clin. Microbiol. Infect., 13(4): 430-433. [Crossref] [PubMed]

11. Liu, Q., Wang, Z.D., Huang, S.Y. and Zhu, X. Q. (2015) Diagnosis of toxoplasmosis and typing of Toxoplasma gondii. Parasit.Vector, 8: 292. [Crossref] [PubMed] [PMC]

12. Salant, H., Spira, D. and Hamburger J.A. (2010) A comparative analysis of coprologic diagnostic methods for detection of Toxoplasma gondii in cats. Am. J. Trop. Med. Hyg., 82(5): 865-870. [Crossref] [PubMed] [PMC]

13. Stojanovic, V. and Foley P. (2011) Infectious disease prevalence in a feral cat population on Prince Edward Island. Canada, Can. Vet. J., 52(9): 979-982. [PubMed] [PMC]

14. Jones, C.D., Okhravi, N., Adamson, P., Tasker, S. and Lightman, S. (2000) Comparison of PCR detection methods for B1, P30 and 18S rDNA genes of Toxoplasma gondii in aqueous humor. Invest. Ophthalmol. Vis. Sci., 41(3): 634-644. [PubMed]

15. Garcia, J.L., Jennari, S.M., Navarro, I.T., Machado, R.Z., Sinhorini, I.L. and Freire, R.L. (2005) Partial protection against tissue cyst formation in pigs vaccinated with crude rhoptry protein of Toxoplasma gondii. Vet. Parasitol., 129(3-4): 209-217. [Crossref] [PubMed]

16. Singh, B. (1997) Molecular methods for diagnosis and epidemiological studies of parasitic infections. Int. J. Parasitol., 27(10): 1135-1145. [Crossref]

17. Orlandi, P.A. and Lampel, K.A. (2000) Extraction- free, filter-based template preparation for rapid and sensitive PCR detection of pathogenic parasitic protozoa. J. Clin. Microbiol., 38(6): 2271-2277. [PubMed] [PMC]

18. Abbasi, I., Branzburg, A., Campos-Ponce, M., Abdel Hafez, S.K., Raoul, F., Craig, P.S. and Hamburger, J. (2003) Coprodiagnosis of Echinococcus granulosis infection of dogs by amplification of a newly identified repeated DNA sequence. Am. J. Trop. Med. Hyg., 69(3): 324-330. [Crossref] [PubMed]

19. Soulsby, E.J.L. (1982) Helminths, Anthropods and Protozoa of Domesticated Animals. 7th ed. The English Language of Book Society and Bailliere Tindal, London. p507-645.

20. Homan, W.L, Vercammen, M., De Braekeleer, J., Verschueren, H. (2000) Identification of a 200- to 300-fold repetitive 529 bp DNA fragment in Toxoplasma gondii, and its use for diagnostic and quantitative PCR. Int. J. Parasitol., 30(1):69-75. [Crossref]

21. Simamora, A.T.A.J., Suratman, N.A. and Apsari, I.A.P. 2015. Isolation and Identification of Toxoplasma gondii oocysts in cat feces in Denpasar with sugar flotation method Sheater. Indo. Med. Vet., 4(2): 88-96.

22. Liang, Y., Chen, J.Q., Meng, Y., Zou, F.C., Hu, J.J. and Esch, G.W. (2016) Occurrence and genetic characterization of GRA6 and SAG2 from Toxoplasma gondii oocysts in cat feces, Kunming, China. Southeast Asian J. Trop. Med. Public Health., 47(6): 1134-1142. [PubMed]

23. Dubey, J.P., Darrington, C., Tiao, N. (2013) Isolation of viable Toxoplasma gondii from tissues and feces of cats from Addis Ababa, Ethiopia. J. Parasitol., 99(1): 56-58. [Crossref] [PubMed]

24. Haddadzadeh, H.R., Khazraiinia, P., Aslani, M., Rezaeian, M., Jamshidi, S., Taheri, M. and Bahonar, A. (2006). Seroprevalence of Toxoplasma gondii infection in stray and household cats in Tehran. Vet. Parasitol., 138(3-4): 211-216. [Crossref] [PubMed]

25. Akhtardanesh, B., Ziaali, N., Sharifi, H. and Hezaei, S.H. (2010) Feline immunodeficiency virus, feline leukemia virus and Toxoplasma gondii in stray and household cats in Kerman-Iran: Seroprevalence and correlation with clinical and laboratory findings. Res. Vet. Sci., 31: 306-310. [Crossref] [PubMed]

26. Dubey, J.P. (1986) Toxoplasmosis in cats. Feline. Pract., 16(4): 12-26.

27. Virgen, J., Castillo, M., Karla, Y., Acosta, V., Eugenia De, S., Guzm'an, M., Matilde Jim'enez, C., Jos'e, C. Segura-Correa, C., Aguilar-Caballero, A.J. and Antonio, P.O. (2012) Prevalence and risk factors of Toxoplasma gondii infection in domestic cats from the tropics of Mexico using serological and molecular tests. Int. Perspect. Infect. Dis., 20: 1-6.

28. Bolais, P.V., Vignoles, P., Pereira, P.F., Keim, R., Arousi, A., Ismail, K., Darde, M.L., Amendoira, M. R. and Mercier, A. (2017) Toxoplasma gondii survey in cats from two environments of the city of Rio de Janeiro, Brazil by modified agglutination test on sera and filter-paper. Parasit. Vectors, 10(1): 88. [Crossref] [PubMed] [PMC]

29. Emily, L.L. and Caroline, D.W. (2013) High prevalence of Toxoplasma gondii oocyst shedding in stray and pet cats (Felis catus) in Virginia, United States. Parasit. Vectors, 6: 266. [Crossref] [PubMed] [PMC]

30. Feitosa, T.F., Vilela, V.L.R., Dantas, E.S., Souto, D.V.O., Pena, H.F.J., Athayde, A.C.R. and Azevedo, S.S. (2014) Toxoplasma gondii and Neospora caninum in domestic cats from the Brazilian semi-arid: Seroprevalence and risk factors. Arq. Bras. Med. Vet. Zootec., 66(4): 1060-1066. [Crossref]

31. Bizhga, B. (2017) Toxoplasmosis under coproscopic diagnosis in cats. Albanian. J. Agric. Sci., 10: 597-603.

32. Gashout, A., Amro, A., Erhuma, M., Al-Dwibe, H., Elmaihub, E., Babba, H., Nattah, N. and Abudher, A. (2016) Toxoplasma gondii in Libya. BMC. Infect. Dis., 16: 157-163. [Crossref] [PubMed] [PMC]

33. Nascimentoa, C.O.M., Silvab, M.L.C.R., Kima, P.C.P., Gomesb, A.A.B., Gomesc, A.L.V., Maiac, R.C.C., Almeidad, J.C. and Motaa, R.A. (2015) Occurrence of Neospora caninum and Toxoplasma gondii DNA in brain tissue from hoary foxes (Pseudalopex vetulus) in Brazil. Acta Trop., 146: 60-65. [Crossref] [PubMed]

34. Franco, C.W.A., de Araujo, F.A.P. and Gennari, S.M. (2013) Toxoplasma gondii in small neotropical wild felids. Braz. J. Vet. Res. Anim. Sci., 50: 50-67.

35. Hartati, S. (2007) The Characterization of SAG1 and R522 Gene, and the Production of SAG1 as Toxoplasma gondii Isolate Local IS-1 Recombinant in Developing Toxoplasmosis Diagnose. Dissertation, Gadjah Mada University, London.

36. Sibley, L.D. and Howe, D.K. (1996) Genetic basis of pathogenicity in toxoplasmosis. In: Gross, U., editor. Toxoplasma gondii. Springer-Verlag, Berlin. p3-15. [Crossref] [PubMed]

37. Ralte, L S., Baidya, J.A., Pandit, S., Jas. R., Nandi, A., Taraphder, S., Isore, D.P., Jana, C. and Ranjan, R. (2017) Detection of Toxoplasma gondii targeting the repetitive microsatellite sequence by PCR. Explor. Anim. Med. Res., 7(2): 159-164.

38. Christina, N., Oury, B., Ambroise, T.P. and Santoro, F. (1991) Restriction fragment length polymorphisms among Toxoplasma gondii strains. Parasitol. Res., 77: 266-268. [Crossref]

39. Parmley, S.F., Gross, U., Sucharczuk, A., Windeck, T., Sgarlato, G.D. and Remington, J.S. (1994) Two alleles of the gene encoding surface antigen p22 in 25 strains of Toxoplasma gondii. J. Parasitol., 80: 293-301. [Crossref] [PubMed]

40. Costa, J. and Bretagnea, S. (2012) Variation of B1 gene and AF146527 repeat element copy numbers according to Toxoplasma gondii strains assessed using real-time quantitative PCR. J. Clin. Microbiol., 50(4): 1452-1454. [Crossref] [PubMed] [PMC]