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Real-time PCR detection of Toxoplasma gondii in surface water samples in São Paulo, Brazil

  • Ana Tereza Galvani
  • Ana Paula Guarnieri Christ
  • José Antonio Padula
  • Mikaela Renata Funada Barbosa
  • Ronalda Silva de Araújo
  • Maria Inês Zanoli Sato
  • Maria Tereza Pepe RazzoliniEmail author
Protozoology - Original Paper

Abstract

Water is considered an important vehicle for the spread of human toxoplasmosis in several countries. Toxoplasma gondii oocysts can persist in the environment for long periods, being highly resistant to the various chemical inactivation processes commonly used by water supply systems, distinctly from simple filtration and flocculation that are efficient in removing oocysts from drinking water. The existing methodologies for identification and quantification of this parasite in water samples are not standardized and have limitations. This study aimed to evaluate the presence of T. gondii oocysts in surface water samples used as a source for the production of drinking water in the State of São Paulo, through the implementation of a specific methodology using real-time PCR technique (qPCR). Volumes of 20 L of the sample were concentrated by filtration in Envirocheck® HV capsules. For DNA extraction, the PowerSoil DNA isolation® kit (currently DNeasy PowerSoil®) was used. The target sequence selected for qPCR was a 62-base-pair fragment of the B1 gene. In the initial recovery evaluation of the method in four replicates of reverse osmosis water, the mean recovery was 48.5% (SD ± 11.5), while the mean recovery for method performance in matrices was 3.2% (SD ± 3.2) (rainy season) and 62.0% (SD ± 6.2) (dry period), suggesting that the characteristics of the samples and the climatic conditions interfere in the recovery efficiency. Of the 39 samples analyzed (May to December 2015), 7.7% (3/39) were positive for T. gondii, and among the ten sources studied; the occurrence of the oocysts was detected in 30% (3/10).

Keywords

Water Toxoplasma gondii B1 gene qPCR 

Notes

Acknowledgements

To Professor Dr. Hilda Fátima de Jesus Pena, Professor Dr. Solange Maria Gennari, and Professor Dr. Rodrigo Martins Soares, from the Laboratory of the Department of Preventive Medicine and Animal Health, School of Veterinary and Zootechnic Medicine, University of Sao Paulo, for their key contributions and donations (Toxoplasma gondi oocysts). Additionally, we are grateful to Professor Dr. Luciana Regina Meireles Jaguaribe Ekman from the Tropical Medicine Institute, University of Sao Paulo, for their substantial contributions in the development of the present study and Dr. Elayse Maria Hachich from the Division of Microbiology and Parasitology of São Paulo State Environmental Company for their support at the beginning of this study.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Ana Tereza Galvani
    • 1
  • Ana Paula Guarnieri Christ
    • 1
  • José Antonio Padula
    • 1
  • Mikaela Renata Funada Barbosa
    • 1
  • Ronalda Silva de Araújo
    • 1
  • Maria Inês Zanoli Sato
    • 1
    • 2
  • Maria Tereza Pepe Razzolini
    • 2
    • 3
    Email author return OK on get
  1. 1.CETESB – São Paulo State Environmental CompanySão PauloBrazil
  2. 2.NARA – Center for Research in Environmental Risk AssessmentSão PauloBrazil
  3. 3.School of Public Health – Environmental Health Department, School of Public HealthUniversity of São PauloSão PauloBrazil

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