Comparison of three diagnostic methods in the diagnosis of cryptosporidiosis and gp60 subtyping of Cryptosporidium parvum in diarrheic calves in Central Anatolia Region of Turkey

Comparison of three diagnostic methods in the diagnosis of cryptosporidiosis and gp60 subtyping of Cryptosporidium parvum in diarrheic calves in Central Anatolia Region of Turkey

The aim of this study was to compare three diagnostic methods for the diagnosis of cryptosporidiosis and to detect subtypes ofCryptosporidium parvum by sequences analyses of gp60 gene in diarrheic calves in several herds in Konya province located in Central Anatolia Region of Turkey. Fecal samples w...

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Main Authors: Yildirim Alparslan, Sevinc Ferda, Onder Zuhal, Duzlu Onder, Ekici Ozlem Derinbay, Isik Nermin, Ciloglu Arif, Simsek Emrah, Yetismis Gamze, Inci Abdullah
Format: Article
Language:English
Published: Sciendo 2021-04-01
Series:The EuroBiotech Journal
Subjects:
cryptosporidium parvum
diarrheic calves
gp60 subtype
nested pcr-rflp
prevalence
taqman qpcr
turkey
Online Access:https://doi.org/10.2478/ebtj-2021-0010
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spelling doaj-0c491aafd31f4a98a21408d94342ec3c2021-09-05T21:00:45ZengSciendoThe EuroBiotech Journal2564-615X2021-04-0152636910.2478/ebtj-2021-0010Comparison of three diagnostic methods in the diagnosis of cryptosporidiosis and gp60 subtyping of Cryptosporidium parvum in diarrheic calves in Central Anatolia Region of TurkeyYildirim Alparslan0Sevinc Ferda1Onder Zuhal2Duzlu Onder3Ekici Ozlem Derinbay4Isik Nermin5Ciloglu Arif6Simsek Emrah7Yetismis Gamze8Inci Abdullah9Erciyes University, Faculty of Veterinary Medicine, Parasitology Department, Kayseri, TurkeySelcuk University, Faculty of Veterinary Medicine, Parasitology Department, Konya, TurkeyErciyes University, Faculty of Veterinary Medicine, Parasitology Department, Kayseri, TurkeyErciyes University, Faculty of Veterinary Medicine, Parasitology Department, Kayseri, TurkeySelcuk University, Faculty of Veterinary Medicine, Parasitology Department, Konya, TurkeySelcuk University, Faculty of Veterinary Medicine, Parasitology Department, Konya, TurkeyErciyes University, Faculty of Veterinary Medicine, Parasitology Department, Kayseri, TurkeyErciyes University, Faculty of Veterinary Medicine, Department of Preclinical Science, Kayseri, TurkeyErciyes University, Faculty of Veterinary Medicine, Parasitology Department, Kayseri, TurkeyErciyes University, Faculty of Veterinary Medicine, Parasitology Department, Kayseri, TurkeyThe aim of this study was to compare three diagnostic methods for the diagnosis of cryptosporidiosis and to detect subtypes ofCryptosporidium parvum by sequences analyses of gp60 gene in diarrheic calves in several herds in Konya province located in Central Anatolia Region of Turkey. Fecal samples were collected from a total of 194 pre-weaned calves (n=158, ≤15 days old, and n=36, 15 to 40 days old), with diarrhoea. For comparative diagnosis, all samples were examined by modified Ziehl-Neelsen staining of fecal smears for the presence of oocyst, nested PCR-RFLP of SSU rRNA and TaqMan qPCR for the detection of Cryptosporidium DNA. A total of 92 (47.4%) and 104 (53.6%) out of the examined samples were found positive by microscopic examination and molecular tools, respectively. The diagnostic sensitivity and specificity of microscopic identification were determined as 88.5% and 100.0%, respectively compared to molecular assays. Cryptosporidium parvum was the only detected species in all positive samples by species-specific qPCR and nested PCR-RFLP assays. Species identifications were further confirmed by sequence analyses of the SSU rRNA PCR products. There was no statistically significant difference in C. parvum prevalence between early pre-weaned calves and calves older than 15 days. The sequence analyses of the gp60 gene of C. parvum isolates revealed a one subtype IIaA13G2R1 belonging to zoonotic family IIa in diarrheic calveshttps://doi.org/10.2478/ebtj-2021-0010cryptosporidium parvumdiarrheic calvesgp60 subtypenested pcr-rflpprevalencetaqman qpcrturkey
collection DOAJ
language English
format Article
sources DOAJ
author Yildirim Alparslan
Sevinc Ferda
Onder Zuhal
Duzlu Onder
Ekici Ozlem Derinbay
Isik Nermin
Ciloglu Arif
Simsek Emrah
Yetismis Gamze
Inci Abdullah
spellingShingle Yildirim Alparslan
Sevinc Ferda
Onder Zuhal
Duzlu Onder
Ekici Ozlem Derinbay
Isik Nermin
Ciloglu Arif
Simsek Emrah
Yetismis Gamze
Inci Abdullah
Comparison of three diagnostic methods in the diagnosis of cryptosporidiosis and gp60 subtyping of Cryptosporidium parvum in diarrheic calves in Central Anatolia Region of Turkey
The EuroBiotech Journal
cryptosporidium parvum
diarrheic calves
gp60 subtype
nested pcr-rflp
prevalence
taqman qpcr
turkey
author_facet Yildirim Alparslan
Sevinc Ferda
Onder Zuhal
Duzlu Onder
Ekici Ozlem Derinbay
Isik Nermin
Ciloglu Arif
Simsek Emrah
Yetismis Gamze
Inci Abdullah
author_sort Yildirim Alparslan
title Comparison of three diagnostic methods in the diagnosis of cryptosporidiosis and gp60 subtyping of Cryptosporidium parvum in diarrheic calves in Central Anatolia Region of Turkey
title_short Comparison of three diagnostic methods in the diagnosis of cryptosporidiosis and gp60 subtyping of Cryptosporidium parvum in diarrheic calves in Central Anatolia Region of Turkey
title_full Comparison of three diagnostic methods in the diagnosis of cryptosporidiosis and gp60 subtyping of Cryptosporidium parvum in diarrheic calves in Central Anatolia Region of Turkey
title_fullStr Comparison of three diagnostic methods in the diagnosis of cryptosporidiosis and gp60 subtyping of Cryptosporidium parvum in diarrheic calves in Central Anatolia Region of Turkey
title_full_unstemmed Comparison of three diagnostic methods in the diagnosis of cryptosporidiosis and gp60 subtyping of Cryptosporidium parvum in diarrheic calves in Central Anatolia Region of Turkey
title_sort comparison of three diagnostic methods in the diagnosis of cryptosporidiosis and gp60 subtyping of cryptosporidium parvum in diarrheic calves in central anatolia region of turkey
publisher Sciendo
series The EuroBiotech Journal
issn 2564-615X
publishDate 2021-04-01
description The aim of this study was to compare three diagnostic methods for the diagnosis of cryptosporidiosis and to detect subtypes ofCryptosporidium parvum by sequences analyses of gp60 gene in diarrheic calves in several herds in Konya province located in Central Anatolia Region of Turkey. Fecal samples were collected from a total of 194 pre-weaned calves (n=158, ≤15 days old, and n=36, 15 to 40 days old), with diarrhoea. For comparative diagnosis, all samples were examined by modified Ziehl-Neelsen staining of fecal smears for the presence of oocyst, nested PCR-RFLP of SSU rRNA and TaqMan qPCR for the detection of Cryptosporidium DNA. A total of 92 (47.4%) and 104 (53.6%) out of the examined samples were found positive by microscopic examination and molecular tools, respectively. The diagnostic sensitivity and specificity of microscopic identification were determined as 88.5% and 100.0%, respectively compared to molecular assays. Cryptosporidium parvum was the only detected species in all positive samples by species-specific qPCR and nested PCR-RFLP assays. Species identifications were further confirmed by sequence analyses of the SSU rRNA PCR products. There was no statistically significant difference in C. parvum prevalence between early pre-weaned calves and calves older than 15 days. The sequence analyses of the gp60 gene of C. parvum isolates revealed a one subtype IIaA13G2R1 belonging to zoonotic family IIa in diarrheic calves
topic cryptosporidium parvum
diarrheic calves
gp60 subtype
nested pcr-rflp
prevalence
taqman qpcr
turkey
url https://doi.org/10.2478/ebtj-2021-0010
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