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Genotyping of Toxoplasma gondii isolates originated from sheep by multilocus PCR-RFLP

Year 2019, , 273 - 280, 14.06.2019

Yılmaz Emre Gencay Sami Gökpınar Cahit Babür Kader Yıldız

https://doi.org/10.33988/auvfd.469526
Cited By: 1

Abstract

The aim of the study was to determine the genotype of Toxoplasma gondii isolates obtained from sheep naturally infected. Genotyping was performed on the sheep isolates (n: 10) using multiplex multilocus nested PCR-RFLP at nine loci (SAG1, SAG2, c22-8, L358, 5’-SAG2, 3’-SAG2, SAG3, BTUB and PK1). Toxoplasma gondii isolates originated from sheep possessed multiple alleles at the same loci. The isolates (n:8) showed alleles of clonal type II and III at two loci (alt. SAG2 and SAG3), type II allele at the locus (c22-8), type I and III alleles at the locus 5'SAG2 and type I and II alleles at the locus L358, type I/III alleles at the locus 3'-SAG2. One isolate showed alleles of type I at four loci (alt. SAG2 and L358), type II allele at the locus c22-8 and type I and II alleles at the locus BTUB, type I/III alleles at the locus 3'-SAG2. One isolate showed alleles of type III at three loci (SAG1, alt. SAG2 and SAG3), type II allele at two loci (c22-8 and L358), type I/III alleles at the locus 3'-SAG2 and type I and II alleles at the locus BTUB. Genotyping may provide knowledge on infection route of T. gondii in people. To the author’s knowledge, it is the first report on genotyping of T. gondii isolates originated from sheep in Turkey. Genotyping of isolates obtained from different hosts and from various regions will contribute to achieving the important knowledge about infection routes and clonal types of T. gondii in Turkey.

Keywords

multilocus nested PCR RFLP Toxoplasma gondii Genotyping sheep Turkey

References

  • Açıcı M, Babür C, Kılıç S ve ark. (2008): Prevalence of antibodies to Toxoplasma gondii infection in humans and domestic animals in Samsun province, Turkey. Trop Anim Health Prod, 40, 311-315.
  • Ajzenberg D, Banuls AL, Tybayrenc M ve ark. (2002): Microsatellite analysis of Toxoplasma gondii shows considerable polymorphism structured into two main clonal groups. Int J Parasitol, 32, 27–38.
  • Aktaş M, Dumanlı N, Babür C ve ark. (2000): Determination of seropositivity for Toxoplasma gondii infection in pregnant and aborted sheep in Elazıg and vicinity by Sabin-Feldman (SF) test. Turk J Vet Anim Sci, 24, 239-241.
  • Araujo F, Slifer T, Kim S (1997): Chronic infection with Toxoplasma gondii does not prevent acute disease or colonization of the brain with tissue cysts following reinfection with different strains of the parasite. J Parasitol, 83, 521–522.
  • Babür C, Karaer Z (1997): Isolation and seroincidence studies of Toxoplasma gondii infection in sheep. Turkiye Parazitol Derg, 21, 293-299.
  • Berger-Schoch AE, Herrmann DC, Schares G ve ark. (2011): Prevalence and genotypes of Toxoplasma gondii in feline faeces (oocysts) and meat from sheep, cattle and pigs in Switzerland. Vet Parasitol, 177, 290–297.
  • Bossi P, Bricaire F (2004): Severe acute disseminated toxoplasmosis. The Lancet, 364, 579.
  • Buxton D, Maley SW, Wright SE ve ark. (2007): Toxoplasma gondii and ovine toxoplasmosis: New aspects of an old story. Vet Parasitol, 149, 25–28.
  • Can H, Doskaya M, Ajzenberg D ve ark. (2014): Genetic characterization of Toxoplasma gondii isolates and toxoplasmosis seroprevalence in stray cats of Izmir, Turkey. PlosOne, 9, e104930.
  • Dao A, Fortier B, Soete M ve ark. (2001): Successful reinfection of chronically infected mice by a different Toxoplasma gondii genotype. Int J Parasitol, 31, 63–65.
  • Döşkaya M, Caner A, Ajzenberg D ve ark. (2013): Isolation of Toxoplasma gondii strains similar to Africa 1 genotype in Turkey. Parasitol Int, 62, 471-474.
  • Dubey JP, (2010): Toxoplasmosis of Animals and Humans. Second edition, CRC Press, Boca Raton.
  • Dubey JP, Zhu XQ, Sundar N ve ark. (2007): Genetic and biological characterization of Toxoplasma gondii isolates of cats from China. Vet Parasitol, 145, 352–356.
  • Dubey JP, Velmurugan GV, Chockalingam A ve ark. (2008): Genetic diversity of Toxoplasma gondii isolate from chickens from Brazil. Vet Parasitol, 157, 299–305.
  • Dumetre A, Ajzenberg D, Rozette L ve ark. (2006): Toxoplasma gondii infection in sheep from Haute-Vienne, France: seroprevalence and isolate genotyping by microsatellite analysis. Vet Parasitol, 142, 376–379.
  • Grigg ME, Ganatra J, Boothroyd JC ve ark. (2001): Unusual abundance of atypical strains associated with human ocular toxoplasmosis. J Infect Dis, 184, 633-639.
  • Halos L, Thébault A, Aubert D ve ark. (2009): An innovative survey underlining the significant level of contamination by Toxoplasma gondii of ovine meat consumed in France. Int J Parasitol, 40, 193–200.
  • Herrmann DC, Pantchev N, Globokar Vrhovec M ve ark. (2010): Atypical Toxoplasma gondii genotypes identified in oocysts shed by cats in Germany. Int J Parasitol, 40, 285–292.
  • Howe DK, Sibley LD (1994): Toxoplasma gondii: analysis of different laboratory stocks of the RH strain reveals genetic heterogeneity. Exp Parasitol, 78, 242-245.
  • Jungersen G, Jensen L, Rask MR ve ark. (2002): Non-lethal infection parameters in mice separate sheep Type II Toxoplasma gondii isolates by virulence. Comp Immunol Microbiol Infect Dis, 25, 187–195.
  • Karakavuk M, Aldemir D, Mercier A ve ark. (2018): Prevalence of toxoplasmosis and genetic characterization of Toxoplasma gondii strains isolated in wild birds of prey and their relation with previously isolated strains from Turkey. PLoS One, 18, e0196159.
  • Khan A, Jordan C, Muccioli C ve ark. (2006): Genetic divergence of Toxoplasma gondii strains associated with ocular toxoplasmosis, Brazil. Emerging Infect Dis, 12, 942–949.
  • Lehmann T, Marcet PL, Graham DH ve ark. (2006): Globalization and the population structure of Toxoplasma gondii. Proc Nat Acad Sci USA, 103, 11423–11428.
  • Mahami-Oskouei M, Moradi M, Fallah E ve ark. (2017): Molecular detection and genotyping of Toxoplasma gondii in chicken, beef, and lamb meat consumed in Northwestern Iran. Iran J Parasitol, 12, 38-45.
  • Mor N, Arslan MO (2007): Seroprevalence of Toxoplasma gondii in sheep in Kars Province. Kafkas Univ Vet Fak Derg, 13, 165-170.
  • Owen MR, Trees AJ (1999): Genotyping of Toxoplasma gondii associated with abortion in sheep. J Parasitol, 85, 382–384.
  • Saeij JP, Boyle JP, Boothroyd JC (2005): Differences among the three major strains of Toxoplasma gondii and their specific interactions with the infected host. Trends Parasitol, 21, 476-481.
  • Saeij JP, Boyle JP, Coller S ve ark. (2006): Polymorphic secreted kinases are key virulence factors in toxoplasmosis. Science, 314, 1780–1783.
  • Satbige AS, Sreekumar C, Rajendran C ve ark. (2017): Isolation and characterization of Toxoplasma gondii from small ruminants (sheep and goats) in Chennai City, South India. J Parasit Dis, 41, 869-873.
  • Scwab EK, Zhu XQ, Majumdar D ve ark. (2014): Geographical patterns of Toxoplasma gondii genetic diversity revealed by multilocus PCR-RFLP genotyping. Parasitol, 141, 453-61.
  • Sevgili M, Babür C, Nalbantoğlu S ve ark. (2005): Determination of seropositivity for Toxoplasma gondii in sheep from Sanlurfa Province. Turk J Vet Anim Sci, 29, 107-111.
  • Sevinc F, Kamburgil K, Dik B ve ark. (2000): The seroprevalence of toxoplasmosis by indirect fluorescent antibody (IFA) test in ewes with and without abortion in Konya province. Firat Univ Saglık Bilim Derg, 14, 137-142.
  • Sibley LD, Boothroyd JC (1992): Virulent strains of Toxoplasma gondii comprise a single clonal lineage. Nature, 359, 82–85.
  • Silva LA, Andrade RO, Carneiro AC ve ark. (2014): Overlapping Toxoplasma gondii genotypes circulating in domestic animals and humans in southeastern Brazil. PLoS One, 9, e90237.
  • Su C, Howe DK, Dubey JP ve ark. (2002): Identification of quantitative trait loci controlling acute virulence in Toxoplasma gondii. Proc Natl Acad Sci USA, 99, 10753–10758.
  • Su C, Zhang X, Dubey JP (2006): Genotyping of Toxoplasma gondii by multilocus PCR-RFLP markers: A high resolution and simple method for identification of parasites. Int J Parasitol, 36, 841–848.
  • Su C, Schwab EK, Zhou P ve ark. (2010): Moving towards an integrated approach to molecular detection and identification of Toxoplasma gondii. Parasitol, 137, 1–11.
  • Velmurugan GV, Dubey JP, Su C (2008): Genotyping studies of Toxoplasma gondii isolates from Africa revealed that the archetypal clonal lineages predominate as in North America and Europe. Vet Parasitol, 155, 314–318.
  • Weiss LM, Kim K (2007): Toxoplasma gondii, The Model Apicomplexan: Perspectives and Methods. Academic Press, Amsterdam.
  • Yıldız K, Babür C, Kılıç S ve ark. (2000): Investigation of anti-Toxoplasma antibodies in slaughtered sheep and cattle as well as in workers in the abattoir of Kırıkkale. Turkiye Parazitol Derg, 24, 180-185.
  • Yildiz K, Kul O, Gokpinar S ve ark. (2014): The relationship between seropositivity and tissue cysts in sheep naturally infected with Toxoplasma gondii. Turk J Vet Anim Sci, 38, 169-175.

Koyun orijinli Toxoplasma gondii izolatlarının multilokus PCR-RFLP yöntemi ile genotiplendirilmesi

Year 2019, , 273 - 280, 14.06.2019

Yılmaz Emre Gencay Sami Gökpınar Cahit Babür Kader Yıldız

https://doi.org/10.33988/auvfd.469526
Cited By: 1

Abstract

Bu çalışmada doğal enfekte koyunlardan elde edilen Toxoplasma gondii izolatlarının (n:10) genotiplendirilmesi amaçlanmıştır. Bu amaçla T. gondii izolatlarının dokuz lokusu (SAG1, SAG2, c22-8, L358, 5’-SAG2, 3’-SAG2, SAG3, BTUB and PK1) multipleks multilokus nested PCR-RFLP yöntemi ile amplifiye edilmiştir. Bunun sonucunda koyun orijinli T. gondii izolatlarının bir dizi lokusta birden çok allel barındırdığı tespit edilmiştir. Koyun izolatları (n: 8) iki lokusta (alt.SAG2 ve SAG3) tip II ve III, bir lokusta (c22-8) klonal tip II, bir lokusta (5'-SAG2) tip I ve III, bir lokusta (L358) I ve II ve bir lokusta (3'-SAG2) ise tip I/III alleli göstermiştir. Bir koyun izolatı; iki lokusta (alt.SAG2 ve L358) tip I, bir lokusta (c22-8) tip II, bir lokusta (BTUB) tip I ve II, bir lokusta (3'-SAG2) ise I/III alleli göstermiştir. Sonuncu koyun izolatı ise üç lokusta (SAG1, alt.SAG2 ve SAG3) tip III, iki lokusta (c22-8 ve L358) tip II, bir lokusta (3'-SAG2) I/III, bir lokusta (BTUB) ise tip I ve II allelleri göstermiştir. Bu çalışma Türkiye’de koyunlarda T. gondii’nin genotiplendirilmesine ilişkin ilk rapordur. Genotiplendirmeye yönelik olarak parazite arakonaklık yapan farklı hayvanlardan ve çeşitli bölgelerden elde edilen izolatlar üzerinde çalışılması Türkiye’de T. gondii’nin bulaşma yolları ve tipleri hakkında önemli bilgiler elde etmeye katkı sağlayacaktır.

Keywords

: Genotiplendirme koyun multilocus nested PCR RFLP Toxoplasma gondii Türkiye

References

  • Açıcı M, Babür C, Kılıç S ve ark. (2008): Prevalence of antibodies to Toxoplasma gondii infection in humans and domestic animals in Samsun province, Turkey. Trop Anim Health Prod, 40, 311-315.
  • Ajzenberg D, Banuls AL, Tybayrenc M ve ark. (2002): Microsatellite analysis of Toxoplasma gondii shows considerable polymorphism structured into two main clonal groups. Int J Parasitol, 32, 27–38.
  • Aktaş M, Dumanlı N, Babür C ve ark. (2000): Determination of seropositivity for Toxoplasma gondii infection in pregnant and aborted sheep in Elazıg and vicinity by Sabin-Feldman (SF) test. Turk J Vet Anim Sci, 24, 239-241.
  • Araujo F, Slifer T, Kim S (1997): Chronic infection with Toxoplasma gondii does not prevent acute disease or colonization of the brain with tissue cysts following reinfection with different strains of the parasite. J Parasitol, 83, 521–522.
  • Babür C, Karaer Z (1997): Isolation and seroincidence studies of Toxoplasma gondii infection in sheep. Turkiye Parazitol Derg, 21, 293-299.
  • Berger-Schoch AE, Herrmann DC, Schares G ve ark. (2011): Prevalence and genotypes of Toxoplasma gondii in feline faeces (oocysts) and meat from sheep, cattle and pigs in Switzerland. Vet Parasitol, 177, 290–297.
  • Bossi P, Bricaire F (2004): Severe acute disseminated toxoplasmosis. The Lancet, 364, 579.
  • Buxton D, Maley SW, Wright SE ve ark. (2007): Toxoplasma gondii and ovine toxoplasmosis: New aspects of an old story. Vet Parasitol, 149, 25–28.
  • Can H, Doskaya M, Ajzenberg D ve ark. (2014): Genetic characterization of Toxoplasma gondii isolates and toxoplasmosis seroprevalence in stray cats of Izmir, Turkey. PlosOne, 9, e104930.
  • Dao A, Fortier B, Soete M ve ark. (2001): Successful reinfection of chronically infected mice by a different Toxoplasma gondii genotype. Int J Parasitol, 31, 63–65.
  • Döşkaya M, Caner A, Ajzenberg D ve ark. (2013): Isolation of Toxoplasma gondii strains similar to Africa 1 genotype in Turkey. Parasitol Int, 62, 471-474.
  • Dubey JP, (2010): Toxoplasmosis of Animals and Humans. Second edition, CRC Press, Boca Raton.
  • Dubey JP, Zhu XQ, Sundar N ve ark. (2007): Genetic and biological characterization of Toxoplasma gondii isolates of cats from China. Vet Parasitol, 145, 352–356.
  • Dubey JP, Velmurugan GV, Chockalingam A ve ark. (2008): Genetic diversity of Toxoplasma gondii isolate from chickens from Brazil. Vet Parasitol, 157, 299–305.
  • Dumetre A, Ajzenberg D, Rozette L ve ark. (2006): Toxoplasma gondii infection in sheep from Haute-Vienne, France: seroprevalence and isolate genotyping by microsatellite analysis. Vet Parasitol, 142, 376–379.
  • Grigg ME, Ganatra J, Boothroyd JC ve ark. (2001): Unusual abundance of atypical strains associated with human ocular toxoplasmosis. J Infect Dis, 184, 633-639.
  • Halos L, Thébault A, Aubert D ve ark. (2009): An innovative survey underlining the significant level of contamination by Toxoplasma gondii of ovine meat consumed in France. Int J Parasitol, 40, 193–200.
  • Herrmann DC, Pantchev N, Globokar Vrhovec M ve ark. (2010): Atypical Toxoplasma gondii genotypes identified in oocysts shed by cats in Germany. Int J Parasitol, 40, 285–292.
  • Howe DK, Sibley LD (1994): Toxoplasma gondii: analysis of different laboratory stocks of the RH strain reveals genetic heterogeneity. Exp Parasitol, 78, 242-245.
  • Jungersen G, Jensen L, Rask MR ve ark. (2002): Non-lethal infection parameters in mice separate sheep Type II Toxoplasma gondii isolates by virulence. Comp Immunol Microbiol Infect Dis, 25, 187–195.
  • Karakavuk M, Aldemir D, Mercier A ve ark. (2018): Prevalence of toxoplasmosis and genetic characterization of Toxoplasma gondii strains isolated in wild birds of prey and their relation with previously isolated strains from Turkey. PLoS One, 18, e0196159.
  • Khan A, Jordan C, Muccioli C ve ark. (2006): Genetic divergence of Toxoplasma gondii strains associated with ocular toxoplasmosis, Brazil. Emerging Infect Dis, 12, 942–949.
  • Lehmann T, Marcet PL, Graham DH ve ark. (2006): Globalization and the population structure of Toxoplasma gondii. Proc Nat Acad Sci USA, 103, 11423–11428.
  • Mahami-Oskouei M, Moradi M, Fallah E ve ark. (2017): Molecular detection and genotyping of Toxoplasma gondii in chicken, beef, and lamb meat consumed in Northwestern Iran. Iran J Parasitol, 12, 38-45.
  • Mor N, Arslan MO (2007): Seroprevalence of Toxoplasma gondii in sheep in Kars Province. Kafkas Univ Vet Fak Derg, 13, 165-170.
  • Owen MR, Trees AJ (1999): Genotyping of Toxoplasma gondii associated with abortion in sheep. J Parasitol, 85, 382–384.
  • Saeij JP, Boyle JP, Boothroyd JC (2005): Differences among the three major strains of Toxoplasma gondii and their specific interactions with the infected host. Trends Parasitol, 21, 476-481.
  • Saeij JP, Boyle JP, Coller S ve ark. (2006): Polymorphic secreted kinases are key virulence factors in toxoplasmosis. Science, 314, 1780–1783.
  • Satbige AS, Sreekumar C, Rajendran C ve ark. (2017): Isolation and characterization of Toxoplasma gondii from small ruminants (sheep and goats) in Chennai City, South India. J Parasit Dis, 41, 869-873.
  • Scwab EK, Zhu XQ, Majumdar D ve ark. (2014): Geographical patterns of Toxoplasma gondii genetic diversity revealed by multilocus PCR-RFLP genotyping. Parasitol, 141, 453-61.
  • Sevgili M, Babür C, Nalbantoğlu S ve ark. (2005): Determination of seropositivity for Toxoplasma gondii in sheep from Sanlurfa Province. Turk J Vet Anim Sci, 29, 107-111.
  • Sevinc F, Kamburgil K, Dik B ve ark. (2000): The seroprevalence of toxoplasmosis by indirect fluorescent antibody (IFA) test in ewes with and without abortion in Konya province. Firat Univ Saglık Bilim Derg, 14, 137-142.
  • Sibley LD, Boothroyd JC (1992): Virulent strains of Toxoplasma gondii comprise a single clonal lineage. Nature, 359, 82–85.
  • Silva LA, Andrade RO, Carneiro AC ve ark. (2014): Overlapping Toxoplasma gondii genotypes circulating in domestic animals and humans in southeastern Brazil. PLoS One, 9, e90237.
  • Su C, Howe DK, Dubey JP ve ark. (2002): Identification of quantitative trait loci controlling acute virulence in Toxoplasma gondii. Proc Natl Acad Sci USA, 99, 10753–10758.
  • Su C, Zhang X, Dubey JP (2006): Genotyping of Toxoplasma gondii by multilocus PCR-RFLP markers: A high resolution and simple method for identification of parasites. Int J Parasitol, 36, 841–848.
  • Su C, Schwab EK, Zhou P ve ark. (2010): Moving towards an integrated approach to molecular detection and identification of Toxoplasma gondii. Parasitol, 137, 1–11.
  • Velmurugan GV, Dubey JP, Su C (2008): Genotyping studies of Toxoplasma gondii isolates from Africa revealed that the archetypal clonal lineages predominate as in North America and Europe. Vet Parasitol, 155, 314–318.
  • Weiss LM, Kim K (2007): Toxoplasma gondii, The Model Apicomplexan: Perspectives and Methods. Academic Press, Amsterdam.
  • Yıldız K, Babür C, Kılıç S ve ark. (2000): Investigation of anti-Toxoplasma antibodies in slaughtered sheep and cattle as well as in workers in the abattoir of Kırıkkale. Turkiye Parazitol Derg, 24, 180-185.
  • Yildiz K, Kul O, Gokpinar S ve ark. (2014): The relationship between seropositivity and tissue cysts in sheep naturally infected with Toxoplasma gondii. Turk J Vet Anim Sci, 38, 169-175.
There are 41 citations in total.

Details

Primary Language Turkish
Subjects Veterinary Surgery
Journal Section Research Article
Authors

Yılmaz Emre Gencay 0000-0002-2154-9663

Sami Gökpınar 0000-0001-7071-869X

Cahit Babür 0000-0002-1961-7305

Kader Yıldız 0000-0001-5802-6156

Publication Date June 14, 2019
Published in Issue Year 2019

Cite

APA Gencay, Y. E., Gökpınar, S., Babür, C., Yıldız, K. (2019). Koyun orijinli Toxoplasma gondii izolatlarının multilokus PCR-RFLP yöntemi ile genotiplendirilmesi. Ankara Üniversitesi Veteriner Fakültesi Dergisi, 66(3), 273-280. https://doi.org/10.33988/auvfd.469526
AMA Gencay YE, Gökpınar S, Babür C, Yıldız K. Koyun orijinli Toxoplasma gondii izolatlarının multilokus PCR-RFLP yöntemi ile genotiplendirilmesi. Ankara Univ Vet Fak Derg. June 2019;66(3):273-280. doi:10.33988/auvfd.469526
Chicago Gencay, Yılmaz Emre, Sami Gökpınar, Cahit Babür, and Kader Yıldız. “Koyun Orijinli Toxoplasma Gondii izolatlarının Multilokus PCR-RFLP yöntemi Ile Genotiplendirilmesi”. Ankara Üniversitesi Veteriner Fakültesi Dergisi 66, no. 3 (June 2019): 273-80. https://doi.org/10.33988/auvfd.469526.
EndNote Gencay YE, Gökpınar S, Babür C, Yıldız K (June 1, 2019) Koyun orijinli Toxoplasma gondii izolatlarının multilokus PCR-RFLP yöntemi ile genotiplendirilmesi. Ankara Üniversitesi Veteriner Fakültesi Dergisi 66 3 273–280.
IEEE Y. E. Gencay, S. Gökpınar, C. Babür, and K. Yıldız, “Koyun orijinli Toxoplasma gondii izolatlarının multilokus PCR-RFLP yöntemi ile genotiplendirilmesi”, Ankara Univ Vet Fak Derg, vol. 66, no. 3, pp. 273–280, 2019, doi: 10.33988/auvfd.469526.
ISNAD Gencay, Yılmaz Emre et al. “Koyun Orijinli Toxoplasma Gondii izolatlarının Multilokus PCR-RFLP yöntemi Ile Genotiplendirilmesi”. Ankara Üniversitesi Veteriner Fakültesi Dergisi 66/3 (June 2019), 273-280. https://doi.org/10.33988/auvfd.469526.
JAMA Gencay YE, Gökpınar S, Babür C, Yıldız K. Koyun orijinli Toxoplasma gondii izolatlarının multilokus PCR-RFLP yöntemi ile genotiplendirilmesi. Ankara Univ Vet Fak Derg. 2019;66:273–280.
MLA Gencay, Yılmaz Emre et al. “Koyun Orijinli Toxoplasma Gondii izolatlarının Multilokus PCR-RFLP yöntemi Ile Genotiplendirilmesi”. Ankara Üniversitesi Veteriner Fakültesi Dergisi, vol. 66, no. 3, 2019, pp. 273-80, doi:10.33988/auvfd.469526.
Vancouver Gencay YE, Gökpınar S, Babür C, Yıldız K. Koyun orijinli Toxoplasma gondii izolatlarının multilokus PCR-RFLP yöntemi ile genotiplendirilmesi. Ankara Univ Vet Fak Derg. 2019;66(3):273-80.

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