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Neospora and neosporosis: achievement and perspectives in diagnosis

100%
Ortega-Mora L. M., Gomez-Bautista M., Fernandez-Garcia A.
Wiadomości Parazytologiczne. Suplement
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2005
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tom 51
2

Diagnosis of bovine neosporosis: Recent advances and perspectives

100%
Ortega-Mora L. M., Fernandez-Garcia A., Gomez-Bautista M.
Acta Parasitologica
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2006
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tom 51
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nr 1
Neospora caninum is considered a major cause of abortion in cattle. Appropriate techniques for diagnosis of bovine neosporosis, both in vivo and in aborted foetuses, have been developed in the last ten years and some of them are commercially available. For diagnosis in live animals, detection of antibodies in serum or milk has been shown to be the best option both at the herd and the individual level. These techniques are excellent tools to examine N. caninum-associated abortion problems and to adopt some basic herd-control measures. Concerning foetal diagnosis, detection of compatible lesions by histological examination and parasites by PCR in brain (as well as heart and liver) are the best choices. Diagnostic criteria to distinguish foetal infection and Neospora-associated abortion are based not only on the demonstration of the parasite in the foetus but also on the extent and severity of the lesions in the foetus, foetal age and the assessment of neosporosis at the herd level. In the near future, new tools to diagnose infection should help to detect animals with parasite reactivation by testing the immune response to stage-specific antigens and lead to the development of molecular typing methods to characterise different parasite isolates. Finally, uniform diagnostic procedures need to be established between laboratories and countries in order to standardise result interpretation. The role of National or Regional Reference Laboratories is essential in countries or regions where control programmes for the disease are being developed.
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Characterisation of NcGRA7 and NcSAG4 proteins: immunolocalisation and their role in the host cell invasion by Neospora caninum tachyzoites

76%
Aguado-Martinez A., Alvarez-Garcia G., Schares G., Risco-Castillo V., Fernandez-Garcia A., Marugan-Hernandez V., Ortega-Mora L. M.
Acta Parasitologica
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2010
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tom 55
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nr 4
304-312
Neospora caninum negatively impacts bovine reproductive performance around the world. Addressing this problem requires a greater understanding of the parasite’s molecular biology. In this study, monoclonal antibodies against recombinant proteins were successfully developed and employed to characterise two different proteins of N. caninum: the acute phase-associated NcGRA7 and the chronic phase-associated NcSAG4. Immunofluorescence with the anti-rNcGRA7 monoclonal antibody suggested that NcGRA7 trafficks from tachyzoite dense granules to the matrix of the parasitophorous vacuole and parasite’s surroundings. Furthermore, NcGRA7 is also expressed in the bradyzoite stage and localised on the matrix of bradyzoite-positive vacuoles. NcGRA7 appears to be partially involved in the tachyzoite-invasion mechanisms, as an anti-rNcGRA7 monoclonal antibody partially inhibited in vitro tachyzoite-invasion. A monoclonal antibody specific for NcSAG4 confirmed this protein’s bradyzoitespecific expression both by western blot and immunofluorescence. However, some bradyzoite-positive vacuoles only weakly expressed NcSAG4, if it was expressed at all. The specificity of the anti-rNcSAG4 monoclonal antibody was confirmed by the recognition of the NcSAG4 in the membrane surface of Nc-1SAG4c transgenic tachyzoites, which constitutively expresses NcSAG4. Blocking NcSAG4 of Nc-1SAG4c tachyzoites with the monoclonal antibody did not affect host cell invasion. However, its implication on the host cell adhesion or host immune evasion should not be discarded.
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