Özlem BOZKURT, ALPARSLAN YILDIRIM, ABDULLAH İNCİ, ARİF ÇİLOĞLU, Zuhal BİŞKİN, ÖNDER DÜZLÜ

Kayseri İli Parklarında Bulunan Oyun Alanlarının Askarit Türleri İle Kontaminasyonunun Parazitolojik ve Moleküler Yöntemlerle Araştırılması

Bu çalışma, Kayseri'de toplam 20 mesire alanı ve parktaki çocuk oyun alanlarından toplanan 248 adet toprak örneğinin köpek ve kedi askarit türleri ile kontaminasyon durumunu saptamak amacıyla yürütülmüştür. Bu amaçla 2009 yılının Ağustos ve Ekim ayları arasında her bir parktan toprak örnekleri tekniğine uygun olarak alınmış ve örnekleme sahası köpek ve kedi dışkılarının varlığı yönünden kontrol edilmiştir. Laboratuvarda toprak ve dışkı örnekleri helmint yumurtalarının varlığı yönünden mikroskobik olarak incelenmiştir. Askarit yumurtası belirlenen örneklerden genomik DNA izolasyonu yapılmıştır. Askarit türlerini belirleme amacıyla elde edilen genomik DNA'lar Toxocara canis, Toxocara cati ve Toxascaris leonina tür spesifik primerler ile PZR analizine tabii tutulmuştur. Mikroskobik analiz neticesinde 20 parkın 10'u (%50.0) ve 248 adet toprak örneğinin 33'ü (%13.3) askarit ve diğer bazı helmint yumurtaları ile kontamine bulunmuştur. En yaygın tür %7.3 ile Toxocara sp. bulunmuş, bunu sırasıyla Toxascaris leonina (%4.0), Spirocerca lupi (%0.8), Taenia sp.Echinococcus sp. (%0.8) ve Ancylostoma caninum (%0.4) izlemiştir. Askarit kontaminasyonu belirlenen toprak örneklerinde moleküler analiz sonuçlarına göre en yaygın tür %12.0 ile T. canis bulunmuş, bunu sırasıyla T. leonina (%7.5) ve T. cati (%3.0) izlemiştir. Türlerin yayılışı arasındaki farklılık istatistiksel açıdan önemli bulunmuştur (P

Investigation of Contamination with Ascarid Species by Parasitological and Molecular Techniques in Playgrounds in Parks of Kayseri Province

This study was designed to determine the contamination status of playgrounds with ascarid species of dogs and cats in Kayseri province of Turkey and was carried out on totally 248 sand samples collected from children playgrounds in 20 picnic areas and parks. For this aim sand samples were collected from each park between August-October 2009 and each sampling area was inspected for the presence of dog and/or cat feces. Sand and feces samples were examined by microscopically for the presence of helmint eggs in the laboratory. Genomic DNA's were extracted from samples that were positive for ascarid eggs. PCR was performed on obtained genomic DNA's with primers specific to Toxocara canis, Toxocara cati and Toxascaris leonina. Ten (50.0%) of 20 parks and 33 (13.3%) of 248 sand samples were found to be contaminated with ascarid and some other helmint eggs. Toxocara sp. was found as the most prevalent species with the ratio of 7.3% and this was followed by T. leonina (4.0%), Spirocerca lupi (0.8%), Taenia sp.- Echinococcus sp. (0.8%) and Ancylostoma caninum (0.4%). Molecular analysis results revealed that T. canis was the most prevalent species with the ratio of 12.0% in the ascarid contaminated sand samples, and this was followed by T. leonina (7.5%) and T. cati (3.0%). The differences among the prevalence rates of species were found statistically significant (P<0.001). Consequently, the molecular prevalence of zoonotic ascarid species was designated in the playgrounds of parks for the first time in Turkey with this study.

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Soulsby EJL: Helminths, Arthropods and Protozoa of Domesticated Animals. Bailliere and Tindall, London, 1986.

Kassai T: Veterinary Helminthology. In, Butterworth-Heinemann, Linarce House, Jordon Hill, pp. 103-108, Oxford, 1999.

Öncel T: İstanbul'da evlerde beslenen köpeklerde Toxocariosis canis. Kafkas Univ Vet Fak Derg, 10 (2): 151-153, 2004.

Kazacos KR: Improved method for recovering ascarid and other helminth eggs from soil associated with epizootics and during survey studies. Am J Vet Res, 44, 896-900, 1983.

Borecka A, Gawor J: Modification of gDNA extraction from soil for PCR designed for the routine examination of soil samples contaminated with Toxocara spp. eggs. J Helminthol, 82 (2): 119-22, 2008.

Graczyk TK, Cranfield MR, Fayer R: Recovery of waterborne oocysts of Cryptosporidium from water samples by the membrane-filter dissolution method. Parasitol Res, 83, 121-125, 1997.

Li MW, Lin RQ, Chen HH: PCR tools for the verification of the specific identity of ascaridoid nematodes from dogs and cats. Mol Cell Probes, 21 (5-6): 349-354, 2007.

Güçlü F, Aydenizöz M: Çocuk parklarındaki kumların köpek ve kedi helminti yumurtaları ile kontaminasyonunun tesbiti. Türkiye Parazitol Derg, 22, 194-198, 1998.

Öge S, Öge H: Prevalence of Toxocara spp. eggs in the soil of public parks in Ankara, Turkey. Dtsch Tierarztl Wschr, 107, 72-75, 2000.

Avcıoğlu H, Burgu A: Seasonal prevalence of Toxocara ova in soil samples from public parks in Ankara, Turkey. Vector Borne Zoonotic Dis, 8 (3): 345-50, 2008.

Toparlak M, Gargılı A, Tüzer E: Contamination of children's play- ground sandpits with Toxocara eggs in İstanbul, Turkey. Turk J Vet Anim Sci, 26, 317-320, 2002.

Kaplan K, Kuk S, Kalkan A: Elazığ'daki çocuk parkları ve oyun sahalarında Toxocara spp. araştırılması. Fırat Üniv Sağlık Bil Derg, 16, 277- , 2002.

Avcıoğlu H, Balkaya I: The relationship of public park accessibility to dogs to the presence of Toxocara species ova in the soil. Vector Borne Zoonotic Dis, 11 (2): 177-180, 2010.

Ayaz E, Yaman M, Gül A: Prevalence of Toxocara spp. eggs in soil of public parks in Van, Turkey. Indian Vet J, 80, 574-576. 2003.

Gürel FS, Ertuğ S, Okyay P: Aydın il merkezindeki parklarda Toxocara spp. yumurta görülme sıklığının araştırılması. Turkiye Parazitol Derg, 29, 179, 2005.

Aydenizöz Özkayhan M: Soil contamination with ascarid eggs in playgrounds in Kırıkkale, Turkey. J Helminthol, 80, 15-18, 2006.

Akdemir C: Visceral larva migrans among children in Kütahya (Turkey) and an evaluation of playgrounds for T. canis eggs. Turk J Pediatr, (2): 158-62, 2010.

Bowles J, Blair D, McManus DP: A molecular phylogeny of the genus Echinococcus. Parasitol, 110, 317-328, 1995.

Blair D, Campos A, Cummings MP, Laclette JP: Evolutionary biology of the parasitic platyhelminths: The role of molecular phylogenetics. Parasitol, 12, 66-71, 1996.

Chilton NB, Gasser RB, Beveridge I: Differences in a ribosomal DNA sequence of the Hypodontus macropi complex. Int J Parasitol, 25, 647-651,

Hoste H, Chilton NB, Gasser RB, Beveridge I: Differences in the second internal transcribed spacer (ribosomal DNA) between five species of Trichostrongylus (Nematode: Trichostrongylidae). Int J Parasitol, 25, 75- , 1995.

Stevenson LA, Chilton NB, Gasser RB: Differentiation of Haemonchus placei from H. contortus (Nematode: Trichostrongylidae) by the ribosomal second internal transcribed spacer. Int J Parasitol, 25, 483-488, 1995.

Jacobs DE, Zhu X, Gasser RB, Chilton NB: PCR-based methods for identification of potentially zoonotic ascaridoid parasites of the dog, fox and cat. Acta Tropica, 68, 191-200, 1997.

Gasser RB, Chilton NB, Hoste H, Stevenson LA: Species identification of trichostrongyle nematodes by PCR-linked RFLP. Int J Parasitol, 24, 291- , 1994.

Gasser RB, Nansen P, Guldberg P: Fingerprinting sequence variation in ribosomal DNA of parasites by DGGE. Mol Cell Probes, 10, 99-105, 1996.

Gasser RB, Stevenson LA, Chilton NB, Nansen P, Bucknell DG, Beveridge I: Species markers for equine strongyles detected in intergenic spacer rDNA by PCRRFLP. Mol Cell Probes, 10, 371-378, 1996.

Fogt-Wyrwas R, Jarozy W, Mızgajska-Wıktor H: Utilizing a poly- merase chain reaction method for the detection of Toxocara canis and T. cati eggs in soil. J Helminthol, 81 (1): 75-8, 2007.