Investigations from Northern Greece on mussels cultivated in areas proximal to wastewaters discharges, as a potential source for human infection with Giardia and Cryptosporidium

Marine bivalves are usually cultivated in shallow, estuarine waters where there is a high concentration of nutrients. Many micro-pollutants, including the protozoan parasites Giardia duodenalis and Cryptosporidium spp., which also occur in such environments, may be concentrated in shellfish tissues during their feeding process. Shellfish can thus be considered as vehicles for foodborne infections, as they are usually consumed lightly cooked or raw. Therefore, the main objective of this study was to investigate the presence of both parasites in Mediterranean mussels, Mytilus galloprovincialis that are cultivated in Thermaikos Gulf, North Greece, which is fed by four rivers that are contaminated with both protozoa. Moreover, the occurrence of these protozoa was monitored in treated wastewaters from 3 treatment plants that discharge into the gulf. In order to identify potential sources of contamination and to estimate the risk for human infection, an attempt was made to genotype Giardia and Cryptosporidium in positive samples. Immunofluorescence was used for detection and molecular techniques were used for both detection and genotyping of the parasites. In total, 120 mussel samples, coming from 10 farms, were examined for the presence of both protozoa over the 6-month farming period. None of them were found positive by immunofluorescence microscopy for the presence of parasites. Only in 3 mussel samples, PCR targeting the GP60 gene detected Cryptosporidium spp. DNA, but sequencing was not successful. Thirteen out of 18 monthly samples collected from the 3 wastewater treatment plants, revealed the presence of Giardia duodenalis cysts belonging to sub-assemblage AII, at relatively low counts (up to 11.2 cysts/L). Cryptosporidium oocysts (up to 0.9 oocysts/L) were also detected in 4 out of 8 samples, although sequencing was not successful at any of the target genes. At the studied location and under the sampling conditions described, mussels tested were not found to be harboring Giardia cysts and the presence of Cryptosporidium was found only in few cases (by PCR detection only). Our results suggest that the likelihood that mussels from these locations act as vehicles of human infection for Giardia and Cryptosporidium seems low.