Parasitic contamination in wastewater and sludge samples in Tunisia using three different detection techniques

The limited availability of water results in the reuse of wastewater or sludge. The Tunisian wastewater regulatory guidelines have specific limits for ova of helminths (<1 egg/l) but none for protozoan parasites. We assessed the presence and loads of parasites in 20 samples of raw, treated wastewater and sludge collected from six wastewater treatment plants. Samples were tested by microscopy using the modified Bailenger method (MBM), immunomagnetic separation (IMS) followed by immunofluorescent assay microscopy, and PCR and sequence analysis for the protozoa Cryptosporidium and Giardia. The seven samples of raw wastewater had a high diversity of helminth and protozoa contamination. Giardia spp., Entamoeba histolytica/dispar, Entamoeba coli, Ascaris spp., Enterobius vermicularis, and Taenia saginata were detected by MBM, and protozoan loads were greater than helminth loads. Cryptosporidium and Giardia were also detected by IMS microscopy and PCR. Six of the eight samples of treated wastewater had parasites: helminths (n = 1), Cryptosporidium (n = 1), Giardia (n = 4), and Entamoeba (n = 4). Four of five samples of sludge had microscopically detectable parasites, and all had both Cryptosporidium and Giardia. The genotypes and subtypes of Cryptosporidium and Giardia were of both human and animal origin. These findings suggest that it may be important to monitor the presence of protozoan parasites in treated wastewater and sludge in Tunisia.