Subtype analysis of Cryptosporidium parvum and Cryptosporidium hominis isolates from humans and cattle in Iran

Nazemalhosseini‐Mojarad, Ehsan; Haghighi, Ali; Taghipour, Niloofar; Keshavarz, Akbar; Mohebi, Seyed Reza; Zali, Mohammad Reza; Xiao, Lihua

doi:10.1016/j.vetpar.2011.01.051

Cited by 47 publications

(40 citation statements)

References 18 publications

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“…C. parvum is the most common in the Middle East, and the prevalence varies between countries and regions. C. hominis has been reported to be more prevalent in Netherlands (12) and Egypt (14), whereas C. parvum in Ireland (24), Ethiopia (13) and Iran (25). The prevalence of the species reported here is similar to that in the UK (9).…”

Section: Discussionsupporting

confidence: 72%

Cryptosporidiosis in Humans with Reference to the First Case of Cryptosporidium hominis Infection in Turkey

Yilmazer¹,

Kucuk²,

Akyıldız³

et al. 2017

Haseki

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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...

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Section: Discussionsupporting

confidence: 72%

Cryptosporidiosis in Humans with Reference to the First Case of Cryptosporidium hominis Infection in Turkey

Yilmazer¹,

Kucuk²,

Akyıldız³

et al. 2017

Haseki

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“…Previous studies in humans in the Middle East, have reported that C. parvum is the predominant Cryptosporidium species in humans (Sulaiman et al, 2005;Meamar et al, 2007;AlBrikan et al, 2008;Pirestani et al, 2008;Hijjawi et al, 2010;Nazemalhosseini-Mojarad et al, 2011;Taghipour et al, 2011;Sharbatkhori et al, 2015), however two studies reported that C.…”

Section: Discussionmentioning

confidence: 97%

“…Few studies have conducted subtyping, but studies in children from urban areas in Iran have reported a predominance of zoonotic C. parvum subtype families (IIa, IId) (Sulaiman et al, 2005;Sharbatkhori et al, 2015;Nazemalhosseini-Mojarad et al, 2011;Taghipour et al, 2011). Analysis of C. parvum and C. hominis isolates from river-water in Iran, showed that all belonged to the IId and Id subtype families, respectively (Mahmoudi et al 2015).…”

Section: Discussionmentioning

confidence: 99%

Genetic characterization of Cryptosporidium in animal and human isolates from Jordan

Hijjawi

Mukbel

Yang

et al. 2016

Veterinary Parasitology

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(Mukbel R). #Joint first authors Highlights First genotyping study of Cryptosporidium in animals in Jordan  Faecal samples (n=284) from domestic animals and humans (n=48) screened  Overall prevalence of 11.6% in animals and six species detected.  Novel Cryptosporidium genotype detected in horses AbstractLittle is known about the epidemiology of Cryptosporidium in Jordan and to date, only one genotyping study has been conducted on Cryptosporidium isolates from Jordanian children. In the present study, a total of 284 faecal samples from Jordanian cattle, sheep, goats and chicken and 48 human faecal samples were screened for the presence of Cryptosporidium using an 18S quantitative PCR (qPCR) and a C. parvum/C. hominis specific qPCR at a lectin locus. Of these, 37 of 284 animal faecal samples were positive by qPCR at the 18S locus giving an overall prevalence of 11.6%. The point prevalence of Cryptosporidium in chickens, sheep, horses, cattle and goats ranged from 4.8% (chickens) to 18.7% (cattle). A total of six species were detected; C. xiaoi (n=9), C.andersoni (n=7), C. ryanae (n=5), C. parvum (n=4), C. baileyi (n=1) and a genetically distinct and potentially novel species in two isolates from horses. Sub-genotype analysis of the 4 C. parvum isolates at the 60-kDa glycoprotein (gp60) locus identified subtype IIaA19G2R1 (n=2) and IIaA16GR1 (n=2). For the human samples, 4 positives (8.3% prevalence) were detected. Of these, two were C. parvum (subtypes IIdA20G1 and IIaA15G2R1) and two were C. hominis (subtypes 1bA9G3 and 1bA10G2R2). Further studies are required to better understand the epidemiology and transmission of Cryptosporidium in Jordan.

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“…Cattle are one of the major hosts of Cryptosporidium parvum, a species that includes subtypes capable of infecting humans (Dixon et al 2011;Meireles et al 2011;Nazemalhosseini-Mojarad et al 2011). Due to their high zoonotic potential, the genotyping of this protozoan is mainly performed using nested PCR and DNA sequencing using various DNA extraction methods and several gene targets (Coklin et al 2010;Diaz et al 2010;Fayer et al 2010a,b;Das et al 2011;Dixon et al 2011;Meireles et al 2011;Nazemalhosseini-Mojarad et al 2011).…”

mentioning

confidence: 99%

“…Due to their high zoonotic potential, the genotyping of this protozoan is mainly performed using nested PCR and DNA sequencing using various DNA extraction methods and several gene targets (Coklin et al 2010;Diaz et al 2010;Fayer et al 2010a,b;Das et al 2011;Dixon et al 2011;Meireles et al 2011;Nazemalhosseini-Mojarad et al 2011). The 18S and GP60 genes are the most commonly analysed genes for Cryptosporidium genotyping (Xiao 2010).…”

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confidence: 99%

Comparison of techniques for DNA extraction and agarose gel staining of DNA fragments using samples of Cryptosporidium

Couto¹,

Sudré²,

Lima³

et al. 2013

Vet. Med. - Czech

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Differentiating between the Cryptosporidium species and their subtypes using only microscopy is impossible. Therefore, molecular tools are indispensable for accurate species and subtype diagnosis. However, if these tools are to be used correctly and accurately, the techniques used must be standardised. In the present study, two molecular techniques for diagnosing Cryptosporidium infection in cows were compared to determine the optimal methods. For each technique, we tested two DNA extraction methods, several annealing temperatures for nested PCR reactions targeting the 18S, SSU rRNA (small subunit ribosomal RNA), and the GP60 (60 kDa glycoprotein) genes, and two types of DNA staining reagents, ethidium bromide and GelRed TM . We determined that one of the tested protocols yields a higher purity of extracted DNA. Additionally, optimised temperatures for the nested PCR of the 18S and GP60 genes were established. Finally, we determined that the GelRed TM dye was more sensitive than ethidium bromide, and its low toxicity facilitates handling and disposal and reduces environmental contamination.

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Subtype analysis of Cryptosporidium parvum and Cryptosporidium hominis isolates from humans and cattle in Iran

Cited by 47 publications

References 18 publications

Cryptosporidiosis in Humans with Reference to the First Case of Cryptosporidium hominis Infection in Turkey

Cryptosporidiosis in Humans with Reference to the First Case of Cryptosporidium hominis Infection in Turkey

Genetic characterization of Cryptosporidium in animal and human isolates from Jordan

Comparison of techniques for DNA extraction and agarose gel staining of DNA fragments using samples of Cryptosporidium

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