Hostname: page-component-8448b6f56d-m8qmq Total loading time: 0 Render date: 2024-04-24T08:15:47.325Z Has data issue: false hasContentIssue false
  • English
  • Français

Isolation and biological and molecular characterization of Neospora caninum (NC-SP1) from a naturally infected adult asymptomatic cattle (Bos taurus) in the state of São Paulo, Brazil

Published online by Cambridge University Press:  11 January 2017

SOLANGE OLIVEIRA ,
RODRIGO MARTINS SOARES ,
JULIANA AIZAWA ,
HERBERT SOUSA SOARES ,
DANIELA PONTES CHIEBAO ,
LUIS MIGUEL ORTEGA-MORA ,
JAVIER REGIDOR-CERRILLO ,
NATÁLIA QUADROS BRESSA SILVA ,
SOLANGE MARIA GENNARI  and
HILDA FÁTIMA JESUS PENA
SOLANGE OLIVEIRA
Affiliation:
Department of Preventive Veterinary Medicine and Animal Health, Faculty of Veterinary Medicine, University of São Paulo, Av. Prof. Dr. Orlando Marques de Paiva, 87, CEP 05508-270, São Paulo, Brazil
RODRIGO MARTINS SOARES
Affiliation:
Department of Preventive Veterinary Medicine and Animal Health, Faculty of Veterinary Medicine, University of São Paulo, Av. Prof. Dr. Orlando Marques de Paiva, 87, CEP 05508-270, São Paulo, Brazil
JULIANA AIZAWA
Affiliation:
Department of Preventive Veterinary Medicine and Animal Health, Faculty of Veterinary Medicine, University of São Paulo, Av. Prof. Dr. Orlando Marques de Paiva, 87, CEP 05508-270, São Paulo, Brazil
HERBERT SOUSA SOARES
Affiliation:
Department of Preventive Veterinary Medicine and Animal Health, Faculty of Veterinary Medicine, University of São Paulo, Av. Prof. Dr. Orlando Marques de Paiva, 87, CEP 05508-270, São Paulo, Brazil
DANIELA PONTES CHIEBAO
Affiliation:
Department of Preventive Veterinary Medicine and Animal Health, Faculty of Veterinary Medicine, University of São Paulo, Av. Prof. Dr. Orlando Marques de Paiva, 87, CEP 05508-270, São Paulo, Brazil
LUIS MIGUEL ORTEGA-MORA
Affiliation:
SALUVET, AnimalHealth Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040 Madrid, Spain
JAVIER REGIDOR-CERRILLO
Affiliation:
SALUVET, AnimalHealth Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040 Madrid, Spain
NATÁLIA QUADROS BRESSA SILVA
Affiliation:
Department of Preventive Veterinary Medicine and Animal Health, Faculty of Veterinary Medicine, University of São Paulo, Av. Prof. Dr. Orlando Marques de Paiva, 87, CEP 05508-270, São Paulo, Brazil
SOLANGE MARIA GENNARI
Affiliation:
Department of Preventive Veterinary Medicine and Animal Health, Faculty of Veterinary Medicine, University of São Paulo, Av. Prof. Dr. Orlando Marques de Paiva, 87, CEP 05508-270, São Paulo, Brazil
HILDA FÁTIMA JESUS PENA*
Affiliation:
Department of Preventive Veterinary Medicine and Animal Health, Faculty of Veterinary Medicine, University of São Paulo, Av. Prof. Dr. Orlando Marques de Paiva, 87, CEP 05508-270, São Paulo, Brazil
*
*Corresponding author: Department of Preventive Veterinary Medicine and Animal Health, Faculty of Veterinary Medicine, University of São Paulo, Av. Prof. Dr. Orlando Marques de Paiva, 87, CEP 05508-270, São Paulo, Brazil. E-mail: hfpena@usp.br
Get access Rights & Permissions [Opens in a new window]

Summary

The biological and genetic diversity of Neospora caninum is very limited because of availability of only a few viable isolates worldwide. This study describes the isolation and biological and molecular characterization of a new viable isolate of N. caninum (NC-SP1), from a cattle in Brazil. Approximately 400 g of brain from a naturally infected adult male cattle from an abattoir was fed to a 2-month-old dog. Neospora-like oocysts were observed on day 7 post-inoculation (PI) and the duration of oocyst shedding was 14 days. The DNA obtained from oocysts was characterized molecularly and the final sequence was 99% identical to homologous sequences of N. caninum available in GenBank®. For bioassay, gerbils (Meriones unguiculatus) were orally inoculated with 10 100 and 1000 oocysts; all gerbils remained clinically normal but developed N. caninum antibodies 14 days PI. Cell culture isolation was successful using the brain homogenate from one of the gerbils and tachyzoites were observed 24 days PI. Microsatellite genotyping revealed a unique genetic profile for this new reference isolate.

Keywords

Neosporosiscattlebioassayoocyst sheddinggerbilsmicrosatellite analysis
Type
Research Article
Information
Parasitology , Volume 144 , Issue 6 , May 2017 , pp. 707 - 711
Check for updates
Copyright
Copyright © Cambridge University Press 2017 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Basso, W., Venturini, L., Venturini, M. C., Hill, D. E., Kwok, O. C., Shen, S. K. and Dubey, J. P. (2001). First isolation of Neospora caninum from the feces of a naturally infected dog. Journal of Parasitology 87, 612–618.CrossRefGoogle ScholarPubMed
Dellarupe, A., Regidor-Cerrillo, J., Jiménez-Ruiz, E., Schares, G., Unzaga, J. M., Venturini, M. C. and Ortega-Mora, L. M. (2014). Comparison of host cell invasion and proliferation among Neospora caninum isolates obtained from oocysts and from clinical cases of naturally infected dogs. Experimental Parasitology 145, 22–28.Google Scholar
Dubey, J. P. (1998). Refinement of pepsin digestion method for isolation of Toxoplasma gondii from infected tissues. Veterinary Parasitology 74, 75–77.Google Scholar
Dubey, J. P. and Schares, G. (2011). Neosporosis in animals – the last five years. Veterinary Parasitology 180, 90–108.Google Scholar
Dubey, J. P., Carpenter, J. L., Speer, C. A., Topper, M. J. and Uggla, A. (1988). Newly recognized fatal protozoan disease of dogs. Journal American Veterinary Medicine Association 192, 1269–1285.Google Scholar
Dubey, J. P., Schares, G. and Ortega-Mora, L. M. (2007). Epidemiology and control of neosporosis and Neospora caninum . Clinical Microbiology Reviews 20, 323–367.Google Scholar
Dubey, J. P., Choudhary, S., Ferreira, L. R., Kwok, O. C. H., Butler, E., Carstensen, M., Yu, L. and Su, C. (2013). Isolation and RFLP genotyping of Toxoplasma gondii from the gray wolf (Canis lupus). Veterinary Parasitology 197, 685–690.Google Scholar
Dubey, J. P., Hemphill, A., Calero-Bernal, R. and Schares, G. (2017). Neosporosis of Animals. CRC Press, Boca Raton, Florida, USA. In press.Google Scholar
Feil, E. J., Li, B. C., Aanensen, D. M., Hanage, W. P. and Spratt, B. G. (2004). eBURST: inferring patterns of evolutionary descent among clusters of related bacterial genotypes from multilocus sequence typing data. Journal of Bacteriology 186, 1518–1530.CrossRefGoogle ScholarPubMed
García-Melo, D. P., Regidor-Cerrillo, J., Ortega-Mora, L. M., Collantes-Fernández, E., de Oliveira, V. S. F., de Oliveira, M. A. P. and da Silva, A. C. (2009). Isolation and biological characterisation of a new isolate of Neospora caninum from an asymptomatic calf in Brazil. Acta Parasitologica 54, 180–185.Google Scholar
Gondim, L. F. P., Pinheiro, A. M., Santos, P. O. M., Jesus, E. E. V., Ribeiro, M. B., Fernandes, H. S., Almeida, M. A. O., Freire, S. M., Meyer, R. and McAllister, M. M. (2001). Isolation of Neospora caninum from the brain of a naturally infected dog, and production of encysted bradyzoites in gerbils. Veterinary Parasitology 101, 1–7.Google Scholar
Gondim, L. F. P., Gao, L. and McAllister, M. M. (2002). Improved production of Neospora caninum oocysts, cyclical oral transmission between dogs and cattle, and in vitro isolation from oocysts. Journal of Parasitology 88, 1159–1163.CrossRefGoogle ScholarPubMed
Gondim, L. F. P., McAllister, M. M. and Gao, L. (2005). Effects of host maturity and prior exposure history on the production of Neospora caninum oocysts by dogs. Veterinary Parasitology 134, 33–39.Google Scholar
Lindsay, D. S., Dubey, J. P. and Duncan, R. B. (1999). Confirmation that the dog is a definitive host for Neospora caninum . Veterinary Parasitology 82, 327–333.CrossRefGoogle ScholarPubMed
Müller, N., Zimmermann, V., Hentrich, B. and Gottstein, B. (1996). Diagnosis of Neospora caninum and Toxoplasma gondii infection by PCR and DNA hybridization immunoassay. Journal of Clinical Microbiology 34, 2850–2852.CrossRefGoogle ScholarPubMed
Pena, H. F. J., Soares, R. M., Ragozo, A. M. A., Monteiro, R. M., Yai, L. E. O., Nishi, S. M. and Gennari, S. M. (2007). Isolation and molecular detection of Neospora caninum from naturally infected sheep from Brazil. Veterinary Parasitology 147, 61–66.Google Scholar
Regidor-Cerrillo, J., Pedraza-Díaz, S., Gómez-Bautista, M. and Ortega-Mora, L. M. (2006). Multilocus microsatellite analysis reveals extensive genetic diversity in Neospora caninum . Journal of Parasitology 92, 517–524.Google Scholar
Regidor-Cerrillo, J., Gómez-Bautista, M., Sodupe, I., Aduriz, G., Álvarez-García, G., Del Pozo, I. and Ortega-Mora, L. M. (2011). In vitro invasion efficiency and intracellular proliferation rate comprise virulence-related phenotypic traits of Neospora caninum . Veterinary Research 42, 41.Google Scholar
Regidor-Cerrillo, J., Díez-Fuertes, F., García-Culebras, A., Moore, D. P., González-Warleta, M., Cuevas, C., Schares, G., Katzer, F., Pedraza-Díaz, S., Mezo, M. and Ortega-Mora, L. M. (2013). Genetic diversity and geographic population structure of bovine Neospora caninum determined by microsatellite genotyping analysis. PLoS ONE 8, 1–12.Google Scholar
Rodrigues, A. A. R., Gennari, S. M., Aguiar, D. M., Sreekumar, C., Hill, D. E., Miska, K. B., Vianna, M. C. B. and Dubey, J. P. (2004). Shedding of Neospora caninum oocysts by dogs fed tissues from naturally infected water buffaloes (Bubalus bubalis) from Brazil. Veterinary Parasitology 124, 139–150.CrossRefGoogle ScholarPubMed
Shwab, E. K., Zhu, X. Q., Majumdar, D., Pena, H. F. J., Gennari, S. M., Dubey, J. P. and Su, C. (2014). Geographical patterns of Toxoplasma gondii genetic diversity revealed by multilocus PCR-RFLP genotyping. Parasitology 141, 453–461.CrossRefGoogle ScholarPubMed
Šlapeta, J. R., Koudela, B., Votýpka, J., Modrý, D., Hořejš, R. and Lukeš, J. (2002). Coprodiagnosis of Hammondia heydorni in dogs by PCR based amplification of ITS 1 rRNA: differentiation from morphologically indistinguishable oocysts of Neospora caninum . Veterinary Journal 163, 147–154.CrossRefGoogle ScholarPubMed
Soares, R. M., Lopes, E. G., Keid, L. B., Sercundes, M. K., Martins, J. and Richtzenhain, L. J. (2011). Identification of Hammondia heydorni oocysts by a heminested-PCR (hnPCR-AP10) based on the H. heydorni RAPD fragment AP10. Veterinary Parasitology 175, 168–172.Google Scholar
Supplementary material: File

Oliveira supplementary material

Oliveira supplementary material

Download Oliveira supplementary material(File)
File 304 KB