Wastewater often contains endocrine disrupting compounds (EDCs) such as ethinyl estradiol (EE2) which is excreted by women who use some oral contraceptive pills. When wastewater treatment is followed by advanced recycling processes, most of the hormone-like EDCs are removed, but it is possible that these very low levels might still adversely affect health. Giving recycled water to mice will allow examination of these health risks but it is first necessary to develop methods to measure extremely low levels of three forms of estrogen (estrone, estradiol and EE2), as well as progesterone and bisphenol A (BPA), an EDC that leaches out of plastic drinking bottles. This research modified clinical ‘ELISA’ tests commonly used to measure hormones in patient blood samples, and established protocols that allowed the sensitive quantification of five EDCs in recycled water.

This research was prompted by concerns that rooftop-harvested rainwater fed into household hot water services might expose the public to harmful pathogens such as salmonella. Householders can be exposed to untreated heated rainwater while washing and when this water is used to wash or prepare food. Water heated to 60°C and maintained at this temperature for several minutes’ kills most, but not all infectious pathogens. The problem is that many Australian hot water services operate at lower temperatures that do not kill enteric pathogens. In this study fewer than 5% of water samples collected from unheated rainwater tanks contained pathogens, but this relatively low frequency still poses a risk and is part of the reason that heated rainwater systems fail to meet existing guidelines for safety. Modifications that would improve safety include installing a device that prevents water cooler than 60°C leaving the storage tank and using cold potable water instead of cold rainwater for tempering or in mixer taps. UV disinfection devices attached to rainwater systems are an effective way to reduce health hazards to acceptable levels.

Cryptosporidium, a microscopic pathogen, forms infectious oocysts which are removed by specific and targeted water treatments. Oocysts can only be seen by using a microscope but finding an infectious dose of 10 oocysts in a litre of water is like finding a needle in a haystack. Usually a larger volume of water, 1000mL, is filtered to recover all the oocysts into a small volume of 0.1 to 1 mL, because this is small enough to be examined under a microscope. It is scientific practice to add some dead, colour-dyed oocysts to the large volume of water. If all the coloured oocysts are counted on the filter, the recovery is 100%. From this it became clear that there was a problem with oocyst recovery. This research found that different elements reduced recovery from different types of water, for example, iron and silica reduced oocyst recovery from river or groundwaters. The approved method for quantifying the environmental occurrence of oocysts can now be modified to increase recovery, and this change improves analysis and consequent management which further reduces risks to public health.

This project developed analytical methods sensitive enough to detect the very low levels of compounds that leach out of old coal tar enamel-lined pipes, then catalogued the chemicals and the levels they were found at in a problematic pipeline. One of the chemicals leaching out of the old lining is probably acted on by microbes to produce another substance with an earthy, musty flavour. None of these were toxic in tests and they are therefore unlikely to pose a risk to human health.

Pathogenic microscopic organisms in source waters pose a risk to public health if water treatment plants do not remove them. It was thought that sensitive PCR tests could be developed to inform decision-making about the most appropriate treatment processes, and to check the absence of pathogens from drinking water. This research focussed on four pathogen classes: cryptosporidium, microcystis, adenovirus and ammonia oxidising bacteria; and evaluated six DNA extraction kits. The Qiagen kit was most cost-effective for extracting DNA and Promega To-Taq polymerase was best for carrying out the PCR test on pathogens in real-world water samples. Other components of the PCR tests that were developed included test controls and DNA standards. A test for each class of pathogen was established and written as a ‘Standard Operating Protocol’ (SOP) which was then applied in different laboratories around Australia. Between-laboratory comparison of results showed the developed PCR tests to be highly reproducible and reliable. They can now be added to the existing suite of tools used to minimise risks to public health.