Kálmán Imre
Banat's University of Agricultural Sciences and Veterinary Medicine, Romania
Title: Comparative detection of Giardia and Cryptosporidium spp. in different water types: microscopy vs. conventional PCR
Biography
Biography: Kálmán Imre
Abstract
Statement of the Problem: Giardia and Cryptosporidium spp. are considered two important waterborne parasites. Their detection in the aquatic environment can be achieved by different methods, each one exhibiting advantages and disadvantages. The aim of the present investigation was to compare the Giardia cysts and Cryptosporidium oocysts water detection efficiency of the immunofluorescence assay (IFA) and the conventional polymerase chain reaction (PCR), in order to bring contribution in the developing and/or improvement of effective identification methods in monitoring of water hygiene for both pathogens. Methodology & Theoretical Orientation: One hundred forty different types of water samples (sewage, river, brook, lake, pond, collector channel) were collected through microfiber filtration, concentrated according to USEPA 1623 protocol and examined for the detection of Giardia and Cryptosporidium spp. through immunofluorescence microscopy and PCR techniques. Findings: Giardia cysts were microscopically detected in 48 (34.3%) samples, while the parasite DNA was recorded in 37 (26.4%) samples. Seven (5.0%) samples showed microscopically positive results for Cryptosporidium oocysts, and the protozoa DNA was isolated from 5 (3.8%) samples. Some of the microscopically positive samples showed negative PCR results and vice-versa, such that the Cohen’s kappa index values, used to measure the degree of agreement between methods, were calculated as being 0.514 for Giardia and 0.826 for Cryptosporidium. Conclusion & Significance: The used methods can be considered effective diagnostic techniques in water detection of Giardia and Cryptosporidium (oo)cysts, and their complementary usage offer a promising tool in the monitoring of aquatic environment pollution for both pathogens.
Acknowledgements: This work was supported by a grant from the Romanian National Authority for Scientific Research and Innovation, CNCS – UEFISCDI, project number PN-II-RU-TE-2014-4-1300.