Toxoplasma gondii and pre-treatment protocols for polymerase chain reaction analysis of milk samples: a field trial in sheep from Southern Italy

  • Alice Vismarra Institute for Experimental Veterinary Medicine of Brescia, Brescia, Italy.
  • Elena Barilli Department of Veterinary Sciences, University of Parma, Parma, Italy.
  • Maura Miceli Department of Veterinary Sciences, University of Parma, Parma, Italy.
  • Carlo Mangia Department of Veterinary Sciences, University of Parma, Parma, Italy.
  • Cristina Bacci | cristina.bacci@unipr.it Department of Veterinary Sciences, University of Parma, Parma, Italy.
  • Franco Brindani Department of Veterinary Sciences, University of Parma, Parma, Italy.
  • Laura Kramer Department of Veterinary Sciences, University of Parma, Parma, Italy.

Abstract

Toxoplasmosis is a zoonotic disease caused by the protozoan Toxoplasma gondii. Ingestion of raw milk has been suggested as a risk for transmission to humans. Here the authors evaluated pre-treatment protocols for DNA extraction on T. gondii tachyzoite-spiked sheep milk with the aim of identifying the method that resulted in the most rapid and reliable polymerase chain reaction (PCR) positivity. This protocol was then used to analyse milk samples from sheep of three different farms in Southern Italy, including real time PCR for DNA quantification and PCR-restriction fragment length polymorphism for genotyping. The pre-treatment protocol using ethylenediaminetetraacetic acid and Tris-HCl to remove casein gave the best results in the least amount of time compared to the others on spiked milk samples. One sample of 21 collected from sheep farms was positive on one-step PCR, real time PCR and resulted in a Type I genotype at one locus (SAG3). Milk usually contains a low number of tachyzoites and this could be a limiting factor for molecular identification. Our preliminary data has evaluated a rapid, cost-effective and sensitive protocol to treat milk before DNA extraction. The results of the present study also confirm the possibility of T. gondii transmission through consumption of raw milk and its unpasteurised derivatives.

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Published
2017-02-09
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Original Articles
Keywords:
Toxoplasma gondii, Serology, Meat juice, ELISA
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How to Cite
Vismarra, A., Barilli, E., Miceli, M., Mangia, C., Bacci, C., Brindani, F., & Kramer, L. (2017). Toxoplasma gondii and pre-treatment protocols for polymerase chain reaction analysis of milk samples: a field trial in sheep from Southern Italy. Italian Journal of Food Safety, 6(1). https://doi.org/10.4081/ijfs.2017.6501

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