Aim:Cryptosporidiosis is a worldwide zoonosis. Microscopic examinations may fail due to indistinctive morphological peculiarities of causative species. Hence, molecular diagnostics has become more important.Methods: Stool samples from 150 patients were examined using carbol-fuchsin stain to determine Cryptosporidium spp. oocysts. Combined nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was used for establishing different species in positive samples. The samples were also screened for other parasites by wet-mount and zinc sulfate flotation methods.Results: Microscopic examinations and molecular techniques revealed 0.67% (1/150) and 8.93% (5/56) positivity, respectively. Nested PCR-RFLP enabled the detection of Cryptosporidium hominis (C. hominis) in one sample, while Cryptosporidium parvum (C. parvum) was detected in four samples. With this study, C. hominis was reported from humans for the first time in Turkey. Among infected ones, three of which were children, four patients excreted C. parvum oocysts had gastroenteritis, and a patient positive for C. hominis had gastroenteritis accompanied by nausea and vomiting. No Giardia spp. and Entamoeba spp. were detected in all infected individuals. Conclusion:C. parvum cases outnumbered C. hominis cases, suggesting a zoonotic transmission although infected individuals were living in an urban area where animal husbandry was not allowed. However, water-borne pathogen contamination in the city's water supply is considered a factor for transmission.Keywords: Cryptosporidiosis, human, Turkey, polymerase chain reaction-restriction fragment length polymorphism Amaç: Cryptosporidiosis dünya çapında bir zoonozdur. Mikroskopik muayene, etken türlerin belirgin olmayan morfolojik özellikleri nedeniyle başarısız olabilir. Bu nedenle moleküler tanı daha da önem kazanmıştır.Yöntemler: Cryptosporidium spp. oosistlerini belirlemek için 150 hastanın dışkı örnekleri karbol-fuksin boya kullanılarak incelendi. Pozitif örneklerde farklı türlerin belirlenmesi için nested polimeraz zincir reaksiyonu-restriksiyon parça uzunluk polimorfizmi (PZR-RPUP) kullanıldı. Örnekler, diğer parazitler için yaş preparasyon ve çinko sülfat flotasyonu yöntemleriyle de araştırıldı. Bulgular:Mikroskobik muayene ve moleküler yöntemlerle sırasıyla %0,67 (1/150) ve %8,93 (5/56) pozitiflik saptandı. Nested PZR-RPUP bir örnekte Cryptosporidium hominis'in (C. hominis), dört örnekte ise Cryptosporidium parvum'un (C. parvum) saptanmasına olanak verdi. Bu çalışma ile C. hominis Türkiye'de ilk kez insanlarda bildirildi. Enfekte olanlar arasında üçü çocuktu, dışkısında C. parvum oosistleri görülen dört hastada gastroenterit, C. hominis pozitif olan bir hastada ise mide bulantısı ve kusmanın eşlik ettiği gastroenterit vardı. Enfekte kişilerin hiçbirinde Giardia spp. ve Entamoeba spp. saptanmadı. Sonuç: C. parvum olgularının C. hominis olgularından fazla olması, enfekte kişilerin hayvancılık yapılmasına izin verilmeyen kentsel alanda yaşamasına rağmen, bir zoonotik bulaşma olduğ...

Objective: Giardiasis is a common disease, and clinical forms can vary based on the assemblage types of the parasite. Detailed information on the subgenotypes may indicate the transmission routes and enlighten the gaps in the epidemiology of the disease. This study aims to reveal the occurrence of giardiasis in Thrace, Turkey, and assemblage types of Giardia duodenalis. Materials and Methods:In total, 573 stool samples taken from the individuals applied to Tekirdağ Central State Hospital in 2009, were examined by wet-mount and zinc sulfate floatation methods. Giardia-positive 26 samples and 64 samples taken from the individuals with gastrointestinal complaints were analyzed by nested PCR-RFLP to differentiate the assemblage types. Sequence analysis was employed for confirmation of assemblage types and subgenotypes.Results: Giardia spp. cysts were detected in 3.66% and 4.54% of the samples with wetmount and zinc sulphate floatation techniques respectively. A total of 27 samples were found positive by nested PCR-RFLP out of 90 samples. Fifteen samples were determined as assemblage A, 2 and 10 samples as B and B/E mix respectively. Sequence analysis showed that the latter assemblage (B/E mix) as A3-B3 mix. Conclusion:Fast identification techniques, namely zinc sulphate flotation can be used for screening stool samples in order to determine Giardia cysts with considerably high sensitivity and specificity. Based on this method, the occurrence rate of giardiasis was found as 4.54% in the studied group. DNA sequencing is necessary to distinguish assemblages and confirm the results of PCR-RFLP.

SummaryThe present study was undertaken to determine time dependent viability changes of purified Cryptosporidium parvum oocysts, stored in antimicrobial-supplemented PBS at +4 o C, using vital dyes (DAPI/PI). The trials demonstrated that vital dyes could provide estimation of oocyst viability, and furthermore, if interpreted correctly, they could be used to determine the degree of the viability in Cryptosporidium parvum oocysts as cell culture-PCR assay is used. Keywords: Cryptosporidium parvum, Viability, Vital dye Cryptosporidium parvum Oocystlerindeki Zamana Bağlı CanlılıkDeğişim Derecesi Vital Boyalarla Belirlenebilir mi? ÖzetBu çalışmada, antimikrobiyal destekli PBS içerisinde +4 o C'de muhafaza edilen pürifiye Cryptosporidium parvum oocystlerindeki canlılık değişimi vital boyalarla (DAPI/PI) belirlenmeye çalışılmıştır. Denemeler, vital boyaların oocyst canlılığındaki değişimi ortaya koyabildiğini ve ayrıca iyi değerlendirildiği taktirde, canlılıktaki söz konusu değişimin derecesinin de, hücre kültürü-PCR uygulamalarına benzer şekilde belirlenebileceğini göstermiştir.