Comparison of Selected Diagnostic Methods for Identification of Cryptosporidium parvum and Cryptosporidium andersoni in Routine Examination of Faeces

2003 ◽  
Vol 50 (8) ◽  
pp. 405-411 ◽  
Author(s):  
M. Kvac ◽  
D. Kvetonova ◽  
G. Puzova ◽  
O. Ditrich
Keyword(s):  
Cryptosporidium Parvum ◽  
Diagnostic Methods ◽  
Routine Examination ◽  
Cryptosporidium Andersoni

Unusual cryptosporidiosis cases in Swedish patients: extended molecular characterization ofCryptosporidium viatorumandCryptosporidiumchipmunk genotype I

Parasitology ◽  
2013 ◽  
Vol 140 (14) ◽  
pp. 1735-1740 ◽  
Author(s):  
MARIANNE LEBBAD ◽  
JESSICA BESER ◽  
MONA INSULANDER ◽  
LILLEMOR KARLSSON ◽  
JENS G. MATTSSON ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Cryptosporidium Parvum ◽  
Geographical Variation ◽  
Indian Subcontinent ◽  
Common Species ◽  
Diagnostic Methods ◽  
Molecular Diagnostic ◽  
Cryptosporidium Hominis ◽  
Genotype I ◽  
Protein Locus

SUMMARYMost human cases of cryptosporidiosis are caused byCryptosporidium parvumorCryptosporidium hominis, but the use of molecular diagnostic methods has revealed that several other less common species or genotypes can also be involved. Here, we describe two unusual causes of cryptosporidiosis, one being the recently described speciesCryptosporidium viatorumand the otherCryptosporidiumchipmunk genotype I. Two Swedish patients who were infected withC. viatorumhad travelled to Kenya and Guatemala, respectively, and two others had been infected withCryptosporidiumchipmunk genotype I in Sweden. None of these four patients were immunocompromised, and all four showed classical symptoms of cryptosporidiosis. We performed extensive molecular characterization, including analysis of four loci. The twoC. viatorumisolates were found to differ slightly at the 70-kDa heat shock protein locus, which may indicate a local geographical variation in this species that has previously been described exclusively on the Indian subcontinent.

scholarly journals Genetic analysis of Cryptosporidium from 2414 humans with diarrhoea in England between 1985 and 2000

2006 ◽  
Vol 55 (6) ◽  
pp. 703-707 ◽  
Author(s):  
F. Leoni ◽  
C. Amar ◽  
G. Nichols ◽  
S. Pedraza-Díaz ◽  
J. McLauchlin
Keyword(s):  
Genetic Analysis ◽  
Cryptosporidium Parvum ◽  
18S Rdna ◽  
Oocyst Wall ◽  
Cryptosporidium Hominis ◽  
Rdna Genes ◽  
Cryptosporidium Andersoni ◽  
Faecal Samples ◽  
Pcr Rflp

The characterization of Cryptosporidium using DNA extracted from whole faecal samples collected from 2414 humans with diarrhoea in England between 1985 and 2000 where cryptosporidial oocysts were detected using conventional methods is described. Characterization was achieved by PCR/RFLP and DNA sequencing of fragments of the Cryptosporidium oocyst wall protein and the 18S rDNA genes. Cryptosporidium parvum was detected in 56.1 % of cases, Cryptosporidium hominis in 41.7 % and a mixture of C. parvum and C. hominis in 0.9 %. In the remainder of cases, Cryptosporidium meleagridis (0.9 %), Cryptosporidium felis (0.2 %), Cryptosporidium andersoni (0.1 %), Cryptosporidium canis (0.04 %), Cryptosporidium suis (0.04 %) and the Cryptosporidium cervine type (0.04 %) were detected.

scholarly journals 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

2021 ◽  
Vol 5 (2) ◽  
pp. 63-69
Author(s):  
Alparslan Yildirim ◽  
Ferda Sevinc ◽  
Zuhal Onder ◽  
Onder Duzlu ◽  
Ozlem Derinbay Ekici ◽  
...  
Keyword(s):  
Cryptosporidium Parvum ◽  
Nested Pcr ◽  
Diagnostic Methods ◽  
Central Anatolia ◽  
Ssu Rrna ◽  
Sequence Analyses ◽  
Significant Difference ◽  
Pcr Rflp ◽  
Species Specific ◽  
Diarrheic Calves

Abstract 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

scholarly journals Cryptosporidiosis in children in a north Jordanian paediatric hospital

2004 ◽  
Vol 10 (4-5) ◽  
pp. 494-501
Author(s):  
E. S. Mahgoub ◽  
A. Al Mahbashi ◽  
B. Abdul Latif
Keyword(s):  
Drinking Water ◽  
Risk Factor ◽  
Cryptosporidium Parvum ◽  
Diagnostic Methods ◽  
Direct Immunofluorescence ◽  
Chemotherapy Treatment ◽  
Paediatric Hospital ◽  
Wet Mount ◽  
Stool Specimens ◽  
Flotation Concentration

We investigated the rate of infection by Cryptosporidium parvum among children from birth to 12 years attending Princess Rahma Teaching Hospital in Irbid, Jordan and evaluated various diagnostic methods. We collected single stool specimens from 300 children; 7 specimens were from children undergoing chemotherapy treatment for cancer. Diagnostic methods used for detection of infection were direct wet mount preparation, flotation concentration, cold Kinyoun Ziehl-Neelsen stain and direct immunofluorescence. We detected C. parvum oocysts in 112 samples [37.3%] using direct immunofluorescence, which showed the highest sensitivity. Source of drinking water appeared to be an important risk factor for transmission of infection. A higher incidence of infection was recorded during January-May, the rainy season

scholarly journals Diagnóstico de criptosporidiose em amostras fecais de bezerros por imunofluorescência direta e microscopia de contraste de fase

2011 ◽  
Vol 41 (6) ◽  
pp. 1057-1062 ◽  
Author(s):  
Weslen Fabricio Pires Teixeira ◽  
William Marinho Dourado Coelho ◽  
Ricardo Velludo Gomes de Soutello ◽  
Fernando Paes de Oliveira ◽  
Camila Guariz Homem ◽  
...  
Keyword(s):  
Cryptosporidium Parvum ◽  
Cryptosporidium Spp ◽  
Cryptosporidium Andersoni

O presente estudo teve como objetivo comparar as técnicas de imunofluorescência direta (IFD) e a microscopia de contraste de fase em solução de Sheather (MCF), para detecção de oocistos de Cryptosporidium spp. em amostras fecais de bezerros. A determinação dos limiares detecção da IFD e da MCF foi realizada utilizando cinco alíquotas de uma amostra fecal de bezerro, comprovadamente negativa para Cryptosporidium spp., adicionadas com diferentes quantidades de oocistos de Cryptosporidium parvum. Ao exame das 5 alíquotas, a IFD e a MCF apresentaram, respectivamente, limiares de detecção de 3,3x104 (duas alíquotas positivas) e 3,3x105 oocistos (1 alíquota positiva) por grama de fezes. Foram também realizadas a comparação entre a positividade obtida e uma análise semiquantitativa do número de oocistos observados por campo de microscopia, em ambos os métodos, em 300 amostras fecais de bezerros. Entre as 300 amostras, 19,7% (59/300) foram positivas pela IFD, com diferença estatisticamente significante (P=0,0098) quando comparada com a positividade obtida pela MCF, que foi de 11,7% (35/300). As amostras positivas foram submetidas à reação em cadeia da polimerase para amplificação de fragmentos da subunidade 18S do rRNA, com posterior sequenciamento dos fragmentos amplificados, o que permitiu a identificação de Cryptosporidium andersoni em 11,9% (7/59) e de C.parvum em 88,1% (52/59) das amostras. Os resultados observados comprovam que a IFD foi mais eficiente que a MCF para detecção de oocistos de Cryptosporidium spp. em amostras fecais de bezerros.

scholarly journals COMPARAÇÃO DA EFICIÊNCIA DAS COLORAÇÕES DE ZIEHL-NEELSEN MODIFICADO E SAFRANINA MODIFICADA NA DETECÇÃO DE OOCISTOS DE Cryptosporidium spp. (EUCOCCIDIORIDA, CRYPTOSPORIDIIDAE) A PARTIR DE AMOSTRAS FECAIS DE BEZERROS DE 0 A 3 MESES

2016 ◽  
Vol 17 (1) ◽  
pp. 119-125
Author(s):  
Renata Dias Rodrigues ◽  
Lara Reis Gomes ◽  
Rafael Rocha de Souza ◽  
Fernando Cristino Barbosa
Keyword(s):  
Cryptosporidium Parvum ◽  
Cryptosporidium Spp ◽  
Cryptosporidium Andersoni

Resumo A criptosporidiose bovina é causada principalmente por quatro espécies distintas: Cryptosporidium parvum, Cryptosporidium bovis, Cryptosporidium ryanae e Cryptosporidium andersoni. A espécie Cryptosporidium parvum (Ordem: Eucoccidiorida, Família: Cryptosporidiidae) é considerada de alto potencial zoonótico, podendo infectar humanos por intermédio da eliminação de oocistos tanto pelos bovinos quanto pelo próprio humano. O objetivo desta pesquisa foi verificar a ocorrência de oocistos de Cryptosporidium spp. em amostras fecais de bezerros (75 machos e 77 fêmeas), tendo sido coletadas 152 amostras de fezes de animais do nascimento até os três meses de idade. O material foi submetido às técnicas de coloração de Ziehl-Neelsen modificado e Safranina modificada, as lâminas foram observadas em toda sua extensão ao microscópio óptico para a verificação da presença de oocistos desta enteroparasitose. Os resultados demonstraram 17,1% (26/152) de positividade no total das amostras examinadas e a análise estatística revelou não haver diferença entre o sexo e as técnicas de coloração utilizadas neste estudo. Conclui-se que a infecção por Cryptosporidium spp. esta presente nas propriedades avaliadas, porém são necessários mais estudos para que o risco de infecção seja mensurado adequadamente e medidas profiláticas implementadas.

scholarly journals Evaluation of recombinant Cryptosporidium hominis GP60 protein and anti-GP60 chicken polyclonal IgY for research and diagnostic purposes

2017 ◽  
Vol 26 (2) ◽  
pp. 205-210 ◽  
Author(s):  
Valéria Chamas Miura ◽  
Sérgio Moraes Aoki ◽  
Paulo Peitl Junior ◽  
Lilian Campos Pires ◽  
Priscila Dalmagro ◽  
...  
Keyword(s):  
Cryptosporidium Parvum ◽  
Diagnostic Methods ◽  
Optimal Conditions ◽  
Cryptosporidium Hominis ◽  
Future Studies ◽  
E Coli ◽  
Gp60 Gene ◽  
Sds Page ◽  
Different Strains ◽  
Temperature Time

Abstract In this study, a method for expressing Cryptosporidium hominis GP60 glycoprotein in Escherichia coli for production of polyclonal anti-GP60 IgY in chickens was developed aiming future studies concerning the diagnosis, prevention and treatment of cryptosporidiosis. The full-length nucleotide sequence of the C. hominis gp60 gene was codon-optimized for expression in E. coli and was synthesized in pET28-a vector. Subcloning was performed on several different strains of BL21 E. coli. Temperature, time and inducer IPTG concentration assays were also performed and analyzed using SDS-PAGE. The optimal conditions were observed at a temperature of 37 °C, with overnight incubation and 1 mM of IPTG. Purification was performed by means of affinity chromatography using the AKTA Pure chromatography system and the Hi-Trap™ HP column (GE Healthcare). The recombinant protein GP60 (rGP60) thus generated was used to immunize laying hens owing the production of polyclonal IgY. Western blot and indirect immunofluorescence showed that the polyclonal antibody was capable of binding to rGP60 and to Cryptosporidium parvum sporozoites, respectively. The rGP60 and the IgY anti-rGP60 generated in this study may be used as templates for research and for the development of diagnostic methods for cryptosporidiosis.

On the Origin of the Microscystalline Structure in Rapidly Solidified IN-100 Powder

1977 ◽  
Vol 35 ◽  
pp. 260-261
Author(s):  
J. M. Walsh ◽  
K. P. Gumz ◽  
J. C. Whittles ◽  
B. H. Kear
Keyword(s):  
The Other ◽  
Coarse Grained ◽  
Microcrystalline Structure ◽  
Routine Examination ◽  
Atomization Process ◽  
Centrifugal Atomization ◽  
Splat Quenching ◽  
Powder Particles ◽  
Dendritic Microstructures ◽  
Rapidly Solidified

During a routine examination of the microstructure of rapidly solidified IN-100 powder, produced by a newly-developed centrifugal atomization process1, essentially two distinct types of microstructure were identified. When a high melt superheat is maintained during atomization, the powder particles are predominantly coarse-grained, equiaxed or columnar, with distinctly dendritic microstructures, Figs, la and 4a. On the other hand, when the melt superheat is reduced by increasing the heat flow to the disc of the rotary atomizer, the powder particles are predominantly microcrystalline in character, with typically one dendrite per grain, Figs, lb and 4b. In what follows, evidence is presented that strongly supports the view that the unusual microcrystalline structure has its origin in dendrite erosion occurring in a 'mushy zone' of dynamic solidification on the disc of the rotary atomizer.The critical observations were made on atomized material that had undergone 'splat-quenching' on previously solidified, chilled substrate particles.

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