Directive 2000/60/EC requires the achievement of a ‘good chemical status’ for surface water within pre-established dates. Disinfection is needed to achieve compulsory final microbial limit values (in Italy for wastewater discharges the parameter Escherichia coli – EC – is imposed by law with a maximum limit value of 5,000 cfu/100 mL). Liquid waste and disinfection by-products must be considered when designing appropriate monitoring of dangerous substances; the specific classes of substances must be investigated according to the typology of received wastewaters and liquid wastes (where applicable) and specific analytical techniques, with Limit of Detection (LOD) lower than the limit values, must be applied; the difficulties faced by national and regional environmental control Agencies is that these techniques have to be applied during ordinary activity and not only for research purposes. The study aims to present the control of dangerous substances, as a screening view, in wastewater treatment plant (WWTP) discharges in the province of Venice (Northern Italy) for the period 2007–2010 based on available data from institutional controls. In addition, the wastewater disinfection process with ozone applied to a medium size WWTP (45,000 Population Equivalents) is presented as a case study, with a view to assessing the microbiological abatement efficacy and the presence of dangerous substances. Discharge quality of the WWTPs in the province of Venice presented mean values that were higher than the LOD, but only for certain metals. For the Paese plant, zinc and chloroform were the only micro-pollutants detected with a higher level than the LOD. From microbiological data in the period 2006–2011 the disinfection abatement efficiency for Paese was, in most cases above 99% for EC, faecal coliform (FC), faecal streptococci (FS) while efficiency was slightly lower for total coliform (TC); however, the proposed criterion aimed at respecting 99.99% abatement was not completely satisfied. Therefore, despite the high organic and industrial load of the considered plant and the need to find an alternative system for chlorine, as chlorine disinfection has been banned in the Veneto region since December 2012, ozone efficiency is not completely satisfactory and other systems such as peracetic or performic acids and UV systems must be considered.