Cryptosporidium parvum in diarrheic calves detected by microscopy and identified by immunochromatographic and molecular methods

Díaz-Lee, A; Mercado, Rubén; Onuoha, Ezenwa Obi; Ozaki, Luiz Shozo; Muñoz, Pamela; Muñoz, Vı́ctor; Martínez, Francisco Javier; Fredes, Fernando

doi:10.1016/j.vetpar.2010.11.001

Cited by 22 publications

(34 citation statements)

References 26 publications

(32 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…This method has been used in many previous studies; Henriksen and Pohlenz (1981), Fathia (1993), Kvac and Vitovec (2003), El-Sherbini and Mohammed (2006), Diaz-Lee et al (2011) and Bhat et al (2012). The MZN staining was the most efficient method in the detection of Cryptosporidium oocysts, so it was recommended as rapid, easy and less costly method for diagnosis of cryptosporidiosis (Abdel-Rady and Sayed, 2008).…”

Section: Discussionmentioning

confidence: 99%

Molecular Characterization of Bubaline Isolate of Cryptosporidium Species from Egypt

Abdelrahma¹,

Megeed²,

Hammam³

et al. 2015

Research J. of Parasitology

View full text Add to dashboard Cite

Cryptosporidium an apicomplexan parasite has the ability to induce diarrhea in bovines, goats, pigs, dogs and cats worldwide. In this study, buffalo calves fecal samples were examined after staining their smears with Modified Ziehl-Neelsen Stain (MZN). Ileal sections were examined for the detection of pathological changes. Further molecular characterization was done using nested PCR amplification and partial sequence analysis. The detected oocysts were morphologically similar to Cryptosporidium parvum. Light microscopic examination of Cryptosporidium infected ileal Tissue Section (TS) stained with H and E revealed the presence of altered mucosal architecture with congestion of blood vessels, infiltration, sloughing and complete erosion of epithelial cells and shortening, blunting, stunting and atrophy of the intestinal villi. Molecular characterization gave PCR amplicons of 18S SSU rRNA gene products approximately at 823 bp. Sequences proved specified generalized relatedness with 21 species of Cryptosporidium but the nucleotide homogeneity percentage was insufficient to designate species or genotypes. Further bioinformatics analysis showed that resulting Cryptosporidium isolates had the closest match with three isolates. It was implied that the Cryptosporidium isolates is mostly like Cryptosporidium parvum (JX237832.1) previously isolated from buffaloes in Ismailia province.

show abstract

Section: Discussionmentioning

confidence: 99%

Molecular Characterization of Bubaline Isolate of Cryptosporidium Species from Egypt

Abdelrahma¹,

Megeed²,

Hammam³

et al. 2015

Research J. of Parasitology

View full text Add to dashboard Cite

show abstract

“…To our knowledge, this study delivered the lowest prevalence rate (3.9%) among other reports from different locations of Turkey [17,[19][20][21][22]24,[26][27][28] . The difference could be due to a vast number of factors such as breed, age, management, environment, and season as well as diagnostic method [5,7,13] . The low prevalence could also be caused by spot fecal sampling instead of serial sampling, which may result in underestimation because of intermittent oocyst excretion [9,11] .…”

Section: Discussionmentioning

confidence: 99%

“…The majority of C. parvum infections appear to be limited to dairy calves under eight weeks of age [10,35] , being highest in calves up to 1-month-old [7,8,36] . In calves, the highest infection rates are reported in calves 7-14 days old [7,37] , 8-14 days old [4,38] and 8-21 days old [39] .…”

Section: Discussionmentioning

confidence: 99%

“…The severity of infection ranges from mild to severe depending on Cryptosporidium species as well as age, previous exposure and immune status of the host. Asymptomatic infection is common in yearling heifers and mature cows [6,7] .…”

Section: Prevalence Of Cryptosporidiosis and Molecular Characterizatimentioning

confidence: 99%

“…The disease in calves has been studied in many countries, with prevalence ranging from 2.4 to 100% [3,7] . In Turkey, cryptosporidiosis was first diagnosed in calves in 1984 [15] .…”

Section: Prevalence Of Cryptosporidiosis and Molecular Characterizatimentioning

confidence: 99%

See 2 more Smart Citations

Erzurum Yöresinde Buzağılarda Cryptosporidiosisin Prevalansı ve Cryptosporidium Türlerinin Moleküler Karekterizasyonu

Güven¹,

Avcıoğlu²,

Balkaya³

et al. 2013

Kafkas Univ Vet Fak Derg

View full text Add to dashboard Cite

SummaryThis study was conducted to determine the prevalence of cryptosporidiosis and to identify Cryptosporidium species found in preweaned calves, in Erzurum, Turkey. Fecal samples were collected from 307 calves up to one month old from 5 dairy farms. Genomic DNA was obtained by DNA extraction (QIAamp DNA Stool kit). The prevalence of cryptosporidiosis was determined based on identification through a nested PCR protocol to amplify fragments of the Cryptosporidium SSU rRNA gene. 3.9% of calves were positive for Cryptosporidium. Calves that were subjected to traditional herd management, were female, aged 2 weeks, and had watery feces were affected by the disease at a greater incidence than those were subjected to planned herd surveillance program, were males, were older than 3 weeks, and had firm feces. DNA sequence analysis of the SSU rRNA gene on all of the PCR positive samples ascertained that C. parvum was the only species present. Further studies should be performed comprehensive fecal analysis for other causative agents for association Cryptosporidium species in calf diarrhea and mortality resulting in economic loss in the region. Keywords: Calf, Cryptosporidium, Nested-PCR, SSU rRNA, Erzurum Erzurum Yöresinde Buzağılarda Cryptosporidiosisin Prevalansı veCryptosporidium Türlerinin Moleküler Karekterizasyonu ÖzetBu çalışma, Erzurum yöresindeki sütten kesim öncesi dönemdeki buzağılarda cryptosporidiosisin prevalansının ve Cryptosporidium türlerinin moleküler karekterizasyonunun ortaya konması amacıyla yapılmıştır. Bu amaçla beş süt işletmesinden, bir aydan küçük 307 buzağının dışkı örnekleri toplanmış ve DNA ekstraksiyonu (QIAamp DNA Stool kit) yapılarak genomik DNA elde edilmiştir. Nested PCR protokolü ile Cryptosporidium SSU rRNA gen bölgesinin kısmi amplifikasyonu yapılmış ve cryptosporidiosis prevalansı %3.9 olarak belirlenmiştir. Geleneksel yöntemlerle yetiştirilen, dişi, 2 haftalık yaşta ve sulu dışkıya sahip buzağıların modern işletmelerde yetiştirilen, erkek, 3 haftadan büyük ve katı kıvamlı dışkıya sahip olanlara göre hastalıktan daha çok etkilendiği saptanmıştır. PCR pozitif örneklerin SSU rRNA gen bölgesi hedef alınarak yapılan DNA dizi analizleri sonuçlarına göre C. parvum'un hayvanlarda bulunan tek tür olduğu anlaşılmıştır. Sonuç olarak, yörede ekonomik kayıplara neden olan buzağı ishalleri ve ölümlerinde Cryptosporidium türleri ile diğer hastalık etkenlerinin etkileşimlerinin ortaya konması amacıyla daha kapsamlı çalışmaların yapılmasının gerekliliği sonucuna varılmıştır.

show abstract

Development of duplex real‐time PCR for quick detection of cryptosporidiosis in goats

Sharma

Kumaresan

Sharma

et al. 2022

Cell Biochemistry & Function

View full text Add to dashboard Cite

Cryptosporidium spp. is the most important foodborne and waterborne pathogens and a leading cause of mortality from foodborne and waterborne gastrointestinal diseases. In neonates of domestic animals, it is associated with consistent diarrhea and dehydration. Cryptosporidium infection begins with the ingestion of sporulated oocytes disseminated by carrier animals that consistently contaminate the environment. Many diagnostic tests are available including microscopy and antigen trap-ELISA, but none of the diagnostic tests available currently cannot differentiate between active and passive infection in the host. In the current study, to address this challenge an mRNA-based duplex TaqMan ® probe PCR was developed to target the Cryptosporidium oocyst wall protein gene and 18SSU rRNA gene in a single tube that can detect metabolically active cryptosporidial oocysts. The mRNA transcripts are the direct indicator of any actively replicating cell and they will help decipher the active stages of its lifecycle in a host. This diagnostic assay was standardized by computing transcript copy number-based limit of detection (LOD). For COWP and 18SSU rRNA genes, the LOD was 7.08 × 10 04 and 5.95 × 10 05 , respectively. During active infections, the oocyst wall protein will be active and so its COWP gene transcripts will act as a marker for active infection. While transcripts for 18SSU rRNA are constitutively expressed in cryptosporidial life cycle. This current diagnostic assay will be a quantitative marker that will help assess the active stages of Cryptosporidium infection in neonates. The disease dynamics will help better understand to formulate the control strategies and contain infection among healthy animals.dead and live oocysts, duplex real-time PCR assay, oocyst wall protein | INTRODUCTIONCryptosporidiosis disease is caused by the intracellular, extracytoplasmic protozoan parasite, which belongs to the phylum Apicomplexa. Cryptosporidiosis is the primary cause of chronic diarrhea among immunocompromised patients, especially in children suffering from HIV, AIDS-defining illness because of its high association with mortality. [1][2][3][4] Cryptosporidium is accountable for 8%-19% of diarrhea in less developed countries, 5 and till the year 2010, it already caused about 100,000 deaths globally. 6 More than 15% of the world's population is drinking contaminated water and suffering from waterborne parasitic protozoan diseases globally, 7 it is responsible for four billion cases of mild to severe diarrhea and 1.6 million deaths annually. 8,9 Cryptosporidium oocysts are

show abstract

Cryptosporidium parvum in diarrheic calves detected by microscopy and identified by immunochromatographic and molecular methods

Cited by 22 publications

References 26 publications

Molecular Characterization of Bubaline Isolate of Cryptosporidium Species from Egypt

Molecular Characterization of Bubaline Isolate of Cryptosporidium Species from Egypt

Erzurum Yöresinde Buzağılarda Cryptosporidiosisin Prevalansı ve Cryptosporidium Türlerinin Moleküler Karekterizasyonu

Development of duplex real‐time PCR for quick detection of cryptosporidiosis in goats

Contact Info

Product

Resources

About