Original Article

The Infestation of Supplied Mincemeat to Sarcocystis Species and Their Polymorphism Using PCR-SSCP Method

Abstract

Background: Sarcocystis spp. is zoonotic parasitic pathogen endangering the safety of meat and derived meat products. This study was carried on for determination of presence of Sarcocystis species in mincemeat in Tabriz (Iran) using single-stranded conformation polymorphism (SSCP) method.

Methods: Thirty samples of mixed ground meat were collected from butchers and DNA was extracted. The 18S rRNA gene fragment of S. cruzi and S. hominis species in the extracted DNA amplified by PCR using one pair specific primers, which resulted in 937 bp length fragments in parasite-positive samples. The results showed that 11 samples out of 30 (36%) were positive for S. cruzi and S. hominis species. The SSCP technique products of gene indicated 4 SSCP patterns (A, B, C and D) among study samples on 18S rRNA gene of S. cruzi and S. hominis.

Results: Contamination with Sarcocystis species in minced meat sold in Tabriz City in Iran is relatively high, which is of particular importance from a health point of view. This study was currently by SSCP technique on minced meat for the first time, based on the primers in the 18S rRNA gene locus, there are polymorphisms in S. cruzi and S. hominis species.

Conclusion: Due to the relatively high level of Sarcocystis contamination in minced meats in Tabriz, the necessary hygienic measures should be must implemented. Also, there was polymorphism in S. cruzi and S. homonis.

1. Fayer R. Sarcocystis spp. in human infections. Clin Microbiol Rev. 2004; 17(4):894–902.
2. .Dubey J, Calero-Bernal R, Rosenthal B, et al. Sarcocystosis of animals and humans: CRC
Press; 2015.
3. Stojecki K, Karamon J, Sroka J, et al. Molecular diagnostics of Sarcocystis spp. infections. Pol J Vet Sci. 15(3): 589–596.
4. Amini-Bavil-Olyaee S, Pourkarim M. How can a novel molecular diagnostic assay instill confidence in researchers and encourage its future use? Hepat Mon. 2012; 12(4): 292–293.
5. Neefs JM, Van de Peer Y, De Rijk P, et al. Compilation of small ribosomal subunit RNA sequences. Nucleic Acids Res.1991; 19 Suppl(Suppl):1987-2015.
6. Kusakabe T, Maekawa K, Ichikawa A, et al. Conformation-selective DNA strand breaks by dynemicin: a molecular wedge into flexible regions of DNA. Biochemistry.1993; 32(43): 11669–11675.
7. Sarafraz N, Spotin A, Haniloo A, et al. Prevalence and molecular analysis of Sarcocystis infections in cattle in Northwest Iran and the first global report of S. gigantea in cattle. Comp Immunol Microbiol Infect Dis. 2020; 73:101566.
8. Najafian HR, Mohebali M, Keshavarz H. Study on frequency of Sarcocystis spp. by macroscopic and microscopic methods in slaughtered cattle in Shahriar district and their public health importance. Pajohesh and Sazandegy, 2008; 77:15–19.
9. Nematollahia A, Khoshkerdar A, Helan J, et al. A study on rate of infestation to Sarcocystis cysts in supplied raw hamburgers. J Parasit Dis. 2015; 39(2): 276–279.
10. Shekarforoush SS, Razavi SM, Farahani, et al. Prevalence of Sarcocysts species in slaughtered goats in Shiraz, Iran. Vet Rec.2005 ; 156(13):418-20.
11. Campos PF, Gilbert TM. DNA extraction from formalin-fixed material. Methods Mol Biol. 2012; 840:81–85.
12. Yang ZQ, Zuo YX, Yao YG, et al. Analysis of the 18SrRNA genes of Sarcocystis species suggests that the morphologically similar organisms from cattle and water buffalo should be considered the same species. Mol Biochem Parasitol. 2001; 115(2): 283-288.
13. Kalantari N, Bayani M, Ghaffari S. Sarcocystis cruzi: First molecular identification from cattle in Iran. Int J Mol Cell Med. 2013; 2(3):125-30.
14. Hamidinejat H, Razi Jalali MH, Nabavi, L. Survey on Sarcocystis infection in slaughtered cattle in South-West of Iran, emphasized on evaluation of muscle squash in comparison with digestion method. J Anim Vet Adv. 2010; 9(12):1724–1726.
15. Hooshyar H, Abbaszadeh Z, Sharafati-Chaleshtori R, et al. Molecular identification of Sarcocystis species in raw hamburgers using PCR-RFLP method in Kashan, central Iran. J Parasit Dis. 2017; 41(4): 1001–1005.
16. Jafari F, Motavallihaghi SM, Bakhtiari M, et al. Sarcocystis bovifelis in raw hamburgers marketed in Hamadan city, western Iran. Iran J Parasitol. 2022; 17(1):36-42.
17. 17.Hajimohammadi B, Ahmadi M, Eslami G, et al. Molecular method development to identify foodborne Sarcocystis hominis in raw beef commercial hamburger. Int J Entric Pathog.2014; 2(4):211-239.
18. Konstantinos KV, Panagiotis P, Antonios VT. PCR–SSCP: A method for the molecular analysis of genetic diseases. Mol Biotechnol.2008; 38(2):155–163.
Files
IssueVol 21 No 1 (2026) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijpa.v21i1.21631
Keywords
Sarcocystis Mincemeat Polymorphism
Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
1.
Nematollahi A, Amandi M, Emdadi M. The Infestation of Supplied Mincemeat to Sarcocystis Species and Their Polymorphism Using PCR-SSCP Method. Iran J Parasitol. 2026;21(1):29-35.