A PCR system for detection of species and genotypes of the Echinococcus granulosus-complex, with reference to the epidemiological situation in eastern Africa

Dinkel, Anke; Njoroge, E.M.; Zimmermann, Anja; Walz, M.; Zeyhle, E.; Elmahdi, I.E.; Mackenstedt, Ute; Römig, Thomas

doi:10.1016/j.ijpara.2003.12.013

Cited by 176 publications

(136 citation statements)

References 27 publications

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“…DNA was extracted using a commercial kit (Roche DNA template extraction kit). The protocol established by Dinkel et al [20] was performed on all DNA samples in order to discriminate with a first screening the G1 strain of E. granulosus from the G5 and G6/7 strains with four different PCR reactions. After amplification, 10 μl of the amplification products were detected and photographed on a 1.5% stained agarose gel.…”

Section: Molecular Studymentioning

confidence: 99%

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Retrospective study on Cystic Echinococcosis in cattle of Italy

Poglayen

Varcasia

Pipia

et al. 2017

J Infect Dev Ctries

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Introduction: Cystic Echinococcosis (CE) is one of the most widespread zoonosis of veterinary and medical importance still constituting a sanitary, economic and socio-cultural problem in Italy. Methodology: The aim of this study was to update epidemiological data on cattle CE in Italy. Data on CE positivity of 5,336 cattle were acquired from abattoir registers between January 2009 and July 2010. Morphobiological characterization of hydatids was performed by direct examination of liver and lungs of 1,664 animals butchered in the same slaughterhouses in 2010. Strain typing of parasites was carried out through the amplification and sequencing of nd1 and cox1 mitochondrial genes. Results: Overall CE prevalence was of 8.1% (430/5,336). Parasitological examination of hydatids showed an overall prevalence of 8.6% with a fertility rate of 0.7% (12/1,664). Regarding localization, hydatids were found in 8% of the livers and in 7.6% of the lungs, respectively. Among positive animals, higher prevalence was observed in the liver (93%) compared to lungs (88.1%) (p > 0.05). Conclusion: The economic loss due to organs condemnation related to CE in cattle amounted to almost € 24,000 per year in the examined abattoir during 2010. Sequence analysis showed the presence of G1 (sheep strain) or Echinococcus granulosus sensu strictu in all examined samples. The G1 confirmed, once more, its possible development into several intermediate hosts such as cattle, especially in areas like southern Italy and Sardinia where the lifecycle of the parasite is still to date carried on by sheep and dogs.

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Section: Molecular Studymentioning

confidence: 99%

“…The molecular surveys carried out both with strain specific PCR through the protocol by Dinkel et al [20] and Mt-DNA sequencing have shown the presence of …”

Section: Parasitological Examinationmentioning

confidence: 99%

Retrospective study on Cystic Echinococcosis in cattle of Italy

Poglayen

Varcasia

Pipia

et al. 2017

J Infect Dev Ctries

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“…Both assays provide the possibility of identifying the ' sheep strain ' of E. granulosus. In addition, 2 research groups have reported PCR-based tests for the identification of several E. granulosus strains (Cabrera et al 2002 ;Dinkel et al 2004). However, these tests have not been further validated for their diagnostic applicability to faecal or environmental samples.…”

Section: Introductionmentioning

confidence: 99%

Identification of taeniid eggs in the faeces from carnivores based on multiplex PCR using targets in mitochondrial DNA

2007

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A multiplex polymerase chain reaction (PCR) was evaluated for the identification of morphologically indistinguishable eggs of the taeniid tapeworms from carnivores using primers targeting mitochondrial genes. The primers for Echinococcus multilocularis (amplicon size 395 bp) were species-specific as assessed by in silico analysis and in the PCR using well-defined control samples. The design of primers that specifically amplify DNA from E. granulosus or Taenia spp. was not possible. The primers designed for E. granulosus also amplified DNA (117 bp) from E. vogeli, and those designed for Taenia spp. amplified products (267 bp) from species of Mesocestoides, Dipylidium and Diphyllobothrium. Nevertheless, as our diagnostic approach includes the concentration of taeniid eggs by sequential sieving and flotation, followed by their morphological detection, this non-specificity has limited practical importance. Sequence analysis of the corresponding amplicon can identify most of the described E. granulosus genotypes. Taenia spp. can be identified by direct sequencing of the 267 bp amplicon, or, for most species, by restriction fragment length polymorphism (RFLP) analysis. The multiplex PCR was readily able to detect 1 egg (estimated to contain 7000 targets, as determined by quantitative PCR). Having been validated using a panel of well-defined samples from carnivores with known infection status, this approach proved to be useful for the identification of taeniid eggs from both individual animals and for epidemiological studies.

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“…This neglected disease is distributed worldwide and causes illnesses in humans and animals (Cardona and Carmena, 2013). Echinococcus ortleppi is found in southern Brazil (Balbinotti et al, 2012) and in determined parts of Europe, Africa, South Asia and the Americas (Dinkel et al, 2004). There are also reports of E. ortleppi causing cystic echinococcosis in humans (Maravilla et al, 2004;Sharma et al, 2013).…”

Section: Resultsmentioning

confidence: 99%

Effect of Poikilacanthus glandulosus (Nees) Ariza branches and gallic acid against viability of Echinococcus ortleppi protoscoleces

Brum¹,

Jesus²,

Belke³

et al. 2017

J App Pharm Sci

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The aim of this study was to assess the antioxidant capacity and to evaluate effect of P. glandulosus on the viability of protoscoleces of Echinococcus spp. as well as determine the possible substance in charge for this action. Echinococcus ortleppi protoscoleces were exposed front of the crude extract and butanol fraction and gallic acid at different times.Phytochemical study revealed the presence of triterpenoids, steroids, coumarins, catechins, phenols, anthocyanins, anthocyanidins and cardioactiveheterosides. The highest polyphenol and flavonoids content was found in the ethyl acetate(151.34±1.71mg/g) and dichlorometane (123.30±1.46mg/g), respectively. For assays with DPPH the ethyl acetate fraction was highlighted (IC5095.48±0.35 µg/mL) and the dichlorometane fraction significantly reduced lipid peroxidation (IC50 135.75±2.39 µg/mL) and also the oxidation of DCFH (15.6 µg/mL). The crude extract, butanol fraction and gallic acid showed scolicidal action of 100% at 15, 10 and 15 minutes, respectively.

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A PCR system for detection of species and genotypes of the Echinococcus granulosus-complex, with reference to the epidemiological situation in eastern Africa

Cited by 176 publications

References 27 publications

Retrospective study on Cystic Echinococcosis in cattle of Italy

Retrospective study on Cystic Echinococcosis in cattle of Italy

Identification of taeniid eggs in the faeces from carnivores based on multiplex PCR using targets in mitochondrial DNA

Effect of Poikilacanthus glandulosus (Nees) Ariza branches and gallic acid against viability of Echinococcus ortleppi protoscoleces

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