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Occurrence of Cryptosporidium and Giardia in recycled waters used for irrigation and first description of Cryptosporidium parvum and C. muris in Greece

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Abstract

Here, we present the first time findings regarding the occurrence of Cryptosporidium and Giardia in sewage waters and the first molecular characterization of Cryptosporidium species in Greece. Biological treatment plants from three regions in Greece have been investigated. The detection of Cryptosporidium oocysts was by modified Ziehl–Neelsen acid fast (MZN-AF) and by immunofluorescence microscopy (IFT) for Cryptosporidium and Giardia (oo)cysts, whereas nested PCR based on the SSU rDNA assay was used for molecular detection of Cryptosporidium followed by sequencing for the genetic characterization of the species. In total, 73 samples (37 raw sewage samples and 38 of treated water samples) were collected and analyzed. Of the 73 water samples, 4 samples were Cryptosporidium-positive by IFT and staining, 12 samples were Cryptosporidium-positive by nested PCR; 9 samples were Giardia-positive by IFT. We showed that Cryptosporidium cysts are found both in the input and the discharge of the biological treatment plants. Molecular characterization of Cryptosporidium based on the small subunit ribosomal DNA gene resulted in the determination of Cryptosporidium parvum and Cryptosporidium muris Greek isolates. This is the first report of Cryptosporidium and Giardia occurrence in wastewaters and the first molecular identification of Cryptosporidium species in Greek environments. As the treated water is used for irrigation, or it is discharged into the sea, our findings indicate that biological treatment facilities constitute a possible risk for public health because the related species are prevalent in humans; the results invite for further epidemiological investigations to evaluate the real public health risk in Greece.

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Acknowledgments

This work was funded by the ARCHIMEDES II project supported by the Greek government.

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The authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. Hellenic Centre for Disease Control and Prevention, 3-5 Agrafon Str., Athens, Greece

    Gregory Spanakos & Anastasia Biba

  2. Department for Parasitology, Entomology and Tropical Diseases, National School of Public Health, 196 Alexandras Ave., Athens, Greece

    Gregory Spanakos & Anastasia Biba

  3. Department of Medical Laboratories, Technological Educational Institution, Agiou Spiridonos, 122 43, Egaleo, Athens, Greece

    Athena Mavridou

  4. Centre for Biomedicine and Infectious Diseases (CBIF), Qinghai Academy for Veterinary Medicine and Animal Sciences, Qinghai University, Xining, 810016, Qinghai, China

    Panagiotis Karanis

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  1. Gregory Spanakos
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  4. Panagiotis Karanis
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Correspondence to Panagiotis Karanis.

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Spanakos, G., Biba, A., Mavridou, A. et al. Occurrence of Cryptosporidium and Giardia in recycled waters used for irrigation and first description of Cryptosporidium parvum and C. muris in Greece. Parasitol Res 114, 1803–1810 (2015). https://doi.org/10.1007/s00436-015-4366-6

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