Neslihan SURSAL, Emrah ŞİMŞEK, Kader YILDIZ

Türkiye’de Bir Kedide Cryptosporidium felis’in Varlığı ve İlk Moleküler Karakterizasyonu

Cryptosporidium türleri, birçok konakta ciddi ishale neden olan oldukça yaygın ve önemli zoonotik protozoan parazitlerdir. Bugüne kadar cryptosporidiosis, Türkiye’de insanlarda ve farklı hayvan konaklarında moleküler olarak rapor edilmiştir. Bu olgu sunumunun amacı, ağır ishalli üç aylık dişi bir kedide mikroskobik olarak saptanan Cryptosporidium ookistlerinin small subunit (SSU) rRNA gen sekansına dayanan moleküler karakterizasyonunu ortaya koymaktır. Cryptosporidium’un SSU rRNA geni, sekans ve filogenetik analizler için amplifiye edilmiştir. Sekans analizi sonucu, Cryptosporidium spp. ookistleri C. felis (izolat adı: ANK_1; erişim numarası: MN394123) olarak karakterize edilmiştir. ANK_1 izolatı, GenBank’ta mevcut C. felis sekanslarının birçoğuyla %99,1 ile %100 arasında benzerlik göstermiştir. Ancak, bazı C. felissekanslarıyla ise yüksek düzeyde genetik farklılık (%1.5-4.3) göstermiştir. Veri setine dahil edilen tüm C. felis izolatlarının SSU rRNA sekansları arasındaki tür içi genetik farklılık %0.0 ile %6.9 arasında belirlenmiştir. Sonuç olarak, bu olgu sunumu ile Türkiye’de bir ev kedisinde C. felis’in varlığı mikroskobik ve moleküler olarak ilk kez gösterilmiştir. Cryptosporidium felis, C. parvum’a kıyasla halk sağlığı açısından nispeten daha az zoonotik risk taşımasına rağmen, insanlar ve kediler arasındaki yakın ilişkiler dikkate alındığında enfekte kedilerin zoonotik bulaşmada önemli olabileceği göz ardı edilmemelidir.

Occurrence and First Molecular Characterization of Cryptosporidium felis in a Cat in Turkey

Cryptosporidium species are highly prevalent and significant zoonotic protozoan parasites that cause severe diarrhea in many hosts. To date, cryptosporidiosis has been molecularly reported from humans and different animal hosts in Turkey. The aim of this case report is to reveal the molecular characterization based on small subunit (SSU) rRNA gene sequence of Cryptosporidium oocysts that were microscopically found in a three-month age female cat with severe diarrhea. SSU rRNA gene of Cryptosporidium was amplified for sequence and phylogenetic analyses. According to sequence analysis, Cryptosporidium spp. oocysts were characterized as C. felis (isolate name: ANK_1; accession number: MN394123). ANK_1 isolate showed a range from 99.1% to 100% identity with the most of C. felis sequences available in GenBank. However, it also exhibited high genetic distance (1.5-4.3%) with some of C. felis sequences. The intraspecific genetic differences among all SSU rRNA sequences of C. felisisolates included in the data set were determined ranged from 0.0% to 6.9%. As a conclusion, we provided the first microscopic and molecular evidence of the occurrence of C. felis in a house cat in Turkey. Although C. felis has a relatively low zoonotic risk to public health when compared with C. parvum, infected cats should not be neglected for zoonotic transmission due to close relationships between cats and humans.

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1. Xiao L, Fayer R, Ryan U, Upton SJ: Cryptosporidium taxonomy: Recent advances and implications for public health. Clin Microbiol Rev, 17 (1): 72- 97, 2004. DOI: 10.1128/cmr.17.1.72-97.2004
2. Fayer R: Taxonomy and species delimitation in Cryptosporidium. Exp Parasitol, 124 (1): 90-97, 2010. DOI: 10.1016/j.exppara.2009.03.005
3. Feng Y, Ryan UM, Xiao L: Genetic diversity and population structure of Cryptosporidium. Trends Parasitol, 34 (11): 997-1011, 2018. DOI: 10.1016/j. pt.2018.07.009
4. Yang R, Ying JLJ, Monis P, Ryan U: Molecular characterisation of Cryptosporidium and Giardia in cats (Felis catus) in Western Australia. Exp Parasitol, 155, 13-18, 2015. DOI: 10.1016/j.exppara.2015.05.001
5. Ryan U, Fayer R, Xiao L: Cryptosporidium species in humans and animals: Current understanding and research needs. Parasitology, 141 (13): 1667-1685, 2014. DOI: 10.1017/S0031182014001085
6. Alves MEM, Martins FDC, Braunig P, Pivoto FL, Sangioni LA, Vogel FSF: Molecular detection of Cryptosporidium spp. and the occurrence of intestinal parasites in fecal samples of naturally infected dogs and cats. Parasitol Res, 117 (9): 3033-3038, 2018. DOI: 10.1007/s00436-018-5986-4
7. Tangtrongsup S, Scorza AV, Reif JS, Ballweber LR, Lappin MR, Salman MD: Seasonal distributions and other risk factors for Giardia duodenalis and Cryptosporidium spp. infections in dogs and cats in Chiang Mai, Thailand. Prev Vet Med, 174: 104820, 2020. DOI: 10.1016/j. prevetmed.2019.104820
8. Li J, Dan X, Zhu K, Li N, Guo Y, Zheng Z, Feng Y, Xiao L: Genetic characterization of Cryptosporidium spp. and Giardia duodenalis in dogs and cats in Guangdong, China. Parasit Vectors, 12 (1): 571, 2019. DOI: 10.1186/s13071-019-3822-z
9. Arai H, Fukuda Y, Hara T, Funakoshi Y, Kaneko S, Yoshida T, Asahi H, Kumada M, Kato K, Koyama T: Prevalence of Cryptosporidium infection among domestic cats in the Tokyo metropolitan district, Japan. Jpn J Med Sci Biol, 43 (1): 7-14, 1990. DOI: 10.7883/yoken1952.43.7
10. Mtambo MM, Nash AS, Blewett DA, Smith HV, Wright S: Cryptosporidium infection in cats: Prevalence of infection in domestic and feral cats in the Glasgow area. Vet Rec, 129 (23): 502-504, 1991. DOI: 10.1136/vr.129.23.502
11.Scorza V, Tangtrongsup S: Update on the diagnosis and management of Cryptosporidium spp infections in dogs and cats. Top Companion Anim Med, 25 (3): 163-169, 2010. DOI: 10.1053/j.tcam.2010.07.007
12. Khurana S, Chaudhary P: Laboratory diagnosis of cryptosporidiosis. Trop Parasitol, 8 (1): 2-7, 2018. DOI: 10.4103/tp.TP_34_17
13. Xiao L, Fayer R: Molecular characterisation of species and genotypes of Cryptosporidium and Giardia and assessment of zoonotic transmission. Int J Parasitol, 38 (11): 1239-1255, 2008. DOI: 10.1016/j.ijpara.2008.03.006
14. Xiao L: Molecular epidemiology of cryptosporidiosis: An update. Exp Parasitol, 124 (1): 80-89, 2010. DOI: 10.1016/j.exppara.2009.03.018
15. Arslan MÖ, İtik Ekinci A: Kars yöresinde sığırlarda Cryptosporidium parvum subtiplerinin belirlenmesi. Kafkas Univ Vet Fak Derg, 18 (Suppl. A): A221-A226, 2012. DOI: 10.9775/kvfd.2012.6565
16. Taylan-Ozkan A, Yasa-Duru S, Usluca S, Lysen C, Ye J, Roellig DM, Feng Y, Xiao L: Cryptosporidium species and Cryptosporidium parvum subtypes in dairy calves and goat kids reared under traditional farming systems in Turkey. Exp Parasitol, 170, 16-20, 2016. DOI: 10.1016/j. exppara.2016.06.014
17. Yildirim A, Adanir R, Inci A, Yukari BA, Duzlu O, Onder Z, Ciloglu A, Simsek E: Prevalence and genotyping of bovine Cryptosporidium species in the Mediterranean and Central Anatolia Region of Turkey. Comp Immunol Microbiol Infect Dis, 69: 101425, 2020. DOI: 10.1016/j. cimid.2020.101425
18. Yılmazer N, Küçük ŞK, Akyıldız G, Gargılı A, Kar S: Cryptosporidiosis in humans with reference to the first case of Cryptosporidium hominis infection in Turkey. Med Bull Haseki, 55: 194-198, 2017. DOI: 10.4274/ haseki.47966
19. Korkmaz UF, Gökpınar S, Yıldız K: Kedilerde bağırsak parazitlerinin yaygınlığı ve halk sağlığı bakımından önemi. Turkiye Parazitol Derg, 40, 194-198, 2016. DOI: 10.5152/tpd.2016.4841
20. Göz Y, Yüksek N, Altug N, Ceylan E, Deger S: Prevalence of Cryptosporidium infection in Van cats. Indian Vet J, 82 (9): 995-996, 2005.
21. Kilinç ÖO, Yilmaz AB, Göz Y, Özkan C, Denizhan V: Determination of Cryptosporidium spp. in Van cats by nested PCR. Med Weter, 74 (7): 456- 459, 2018. DOI: 10.21521/mw.6008
22. Purina: Fecal scoring chart 2016. https://www.proplanveterinarydiets. ca/wp-content/uploads/2016/04/PPPVD-Fecal-Scoring-Chart-EN-FINAL. pdf; Accessed: 12.02.2020.
23. Heine J: A simple technic for the demonstration of cryptosporidia in feces. Zentralbl Veterinarmed B, 29 (4): 324-327, 1982.
24. Ryan U, Xiao L, Read C, Zhou L, Lal AA, Pavlasek I: Identification of novel Cryptosporidium genotypes from the Czech Republic. Appl Environ Microbiol, 69 (7): 4302-4307, 2003. DOI: 10.1128/aem.69.7.4302- 4307.2003
25. Kumar S, Stecher G, Li M, Knyaz C, Tamura K: MEGA X: Molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol, 35 (6): 1547-1549, 2018. DOI: 10.1093/molbev/msy096
26. Kimura M: A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol, 16 (2): 111-120, 1980. DOI: 10.1007/BF01731581
27. McReynolds CA, Lappin MR, Ungar B, McReynolds LM, Bruns C, Spilker MM, Thrall MA, Reif JS: Regional seroprevalence of Cryptosporidium parvum-specific IgG of cats in the United States. Vet Parasitol, 80 (3): 187-195, 1999. DOI: 10.1016/s0304-4017(98)00219-2
28.Rambozzi L, Menzano A, Mannelli A, Romano S, Isaia MC: Prevalence of cryptosporidian infection in cats in Turin and analysis of risk factors. J Feline Med Surg, 9 (5): 392-396, 2007. DOI: 10.1016/j.jfms.2007.03.005
29. Lucio-Forster A, Griffiths JK, Cama VA, Xiao L, Bowman DD: Minimal zoonotic risk of cryptosporidiosis from pet dogs and cats. Trends Parasitol, 26 (4): 174-179, 2010. DOI: 10.1016/j.pt.2010.01.004
30. Kváč M, Hofmannová L, Ortega Y, Holubová N, Horčičková M, Kicia M, Hlásková L, Květoňová D, Sak B, McEvoy J: Stray cats are more frequently infected with zoonotic protists than pet cats. Folia Parasitol, 64: 034, 2017. DOI: 10.14411/fp.2017.034
31. Ito Y, Itoh N, Kimura Y, Kanai K: Molecular detection and characterization of Cryptosporidium spp. among breeding cattery cats in Japan. Parasitol Res, 115 (5): 2121-2123, 2016. DOI: 10.1007/s00436- 016-4984-7
32. Bednarska M, Jankowska I, Pawelas A, Piwczynska K, Bajer A, Wolska-Kusnierz B, Wielopolska M, Welc-Faleciak R: Prevalence of Cryptosporidium, Blastocystis, and other opportunistic infections in patients with primary and acquired immunodeficiency. Parasitol Res, 117 (9): 2869- 2879, 2018. DOI: 10.1007/s00436-018-5976-6
33. Beser J, Toresson L, Eitrem R, Troell K, Winiecka-Krusnell J, Lebbad M: Possible zoonotic transmission of Cryptosporidium felis in a household. Infect Ecol Epidemiol, 5 (1): 28463, 2015. DOI: 10.3402/iee.v5.28463
34. Raccurt CP: [Human Cryptosporidium felis infection in the world]. Parasite, 14 (1): 15-20, 2007. DOI: 10.1051/parasite/2007141015
35. Ebner J, Koehler AV, Robertson G, Bradbury RS, Jex AR, Haydon SR, Stevens MA, Norton R, Joachim A, Gasser RB: Genetic analysis of Giardia and Cryptosporidium from people in Northern Australia using PCR-based tools. Infect Genet Evol, 36, 389-395, 2015. DOI: 10.1016/j.meegid. 2015.08.034
36. Elwin K, Hadfield SJ, Robinson G, Chalmers RM: The epidemiology of sporadic human infections with unusual cryptosporidia detected during routine typing in England and Wales, 2000-2008. Epidemiol Infect, 140 (4): 673-683, 2012. DOI: 10.1017/S0950268811000860