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This study investigated the phytotoxicity effect of untreated and treated wastewater collected from two different configurations of domestic wastewater treatment plants in South Africa. The phytotoxicity effect on vegetable seed growth was studied in terms of germination index (GI), relative seed germination (RSG) and relative root elongation (RRE) using four commercial crop varieties, viz., tomato (Lycopersicon esculentum), radish (Raphanus sativus), carrot (Daucus carota) and onion (Allium cepa). According to phyototoxicity limits, 80% germination and above is regarded as non-toxic and less than 50% GI is regarded as highly toxic and not suitable for agricultural purposes. In our study, seeds were irrigated with concentrations of 25%, 50%, 75%, 100% of treated effluent (TE) and untreated effluent (UTE). The TE results were best with the highest GI (%) recorded as tomato, 177; carrot, 158.5; onion, 132; and lettuce, 124. The results of this study indicate that TE showed no phytotoxicty effects and recorded above 80% GI. The UTE irrigated crops reached a GI of only 50% and above which is clear evidence of the beneficial effect of waste water treatment. The overall results confirmed that treated wastewater has a beneficial effect on agricultural crops and can be used as a liquid fertilizer. [ABSTRACT FROM AUTHOR]

South Africa is a water-scarce country with increasing pressure on its water resources. Urgent interventions are needed to protect water security within this rapidly developing country. This paper reports on an interdisciplinary Water Security Colloquium, convened by the South African Young Academy of Science in 2014. A selected group of water professionals from academia, civil society and local government was brought together to discuss water security under three focus themes: 'public expectations and municipal obligations', 'water security and governance: challenges and advances', and 'water re-use: health and infrastructural considerations'. Participant perceptions were generated using a focus group methodology, combined with participatory data collection methods. Under each theme, inputs were categorised as 'challenges', 'gaps in knowledge', and 'solutions/recommendations' and these inputs were thereafter ranked in order of importance via a 'voting' process. Major challenges perceived included a lack of both skills and political will in government, a need to restore citizen trust in government intention and capability to deliver water-related services, and a failure to up-scale existing water re-use technology. Participants identified understanding of the process and implications of the Green and Blue Drop Programmes, knowledge transfer to the public, and the role of educators as major knowledge gaps. The top suggestions proposed included creating public awareness around and buy-in to initiatives to improve water security, accessible and user-friendly conversion of research results to implementation, and ensuring an active role for educators in creating awareness around water security. In view of the concerns identified, participants suggested as potential solutions: improving government and public understanding around water issues, incentivising water re-use and conservation, introducing rising block tariffs and improving human capacity development in the water sector. Developing the ecological infrastructure that protects both quantity and quality of water and building strong partnerships among all stakeholders were also recognised as key. [ABSTRACT FROM AUTHOR]

Three wastewater treatment plants in South Africa were investigated to understand the phylogeny and distribution of Microthrix parvicella using real-time polymerase chain reaction (RT-PCR). The phylogenetic analysis of the 16S rRNA of M. parvicella revealed 98% to 100% homology of South African clones to M. parvicella reported in Genbank. The standard curves for RT-PCR showed R2 values greater than 0.99, accurate for quantification. The relative occurrence of M. parvicella 16S rRNA gene copies in the three wastewater treatment plants was in the range 0% to 3.97%. M. parvicella copies increased when the environmental temperature (≤20°C) and food/microorganism (F/M) ratio was low. The M. parvicella 16S rRNA copies could be positively correlated to the sludge volume index at low temperature. At higher temperature, there was a rapid reduction in M. parvicella population irrespective of other favorable factors, indicating the strong influence of temperature on filamentous proliferation. RT-PCR has potential applications in wastewater treatment plants to monitor sudden shift in the microbial population and assessing the plants efficacy. [ABSTRACT FROM AUTHOR]

Infections caused by protozoan parasites are a major public health concern globally. These infections are commonly diagnosed during water-borne outbreaks, necessitating accurate and highly sensitive detection procedures to assure public health protection. Current molecular techniques are challenged by several factors, such as low parasite concentration, inefficient DNA extraction methods, and inhibitors in environmental samples. This study focused on the development and validation of a molecular protocol for DNA extraction, efficient protozoan (oo)cyst recovery and quantification of protozoan parasites from wastewater using droplet digital polymerase chain reaction (ddPCR). Five DNA extraction methods, including commercial kits, custom phenol-chloroform, and in-house modified methods, were evaluated. The efficiency of each method was assessed via spectrophotometric analysis and ddPCR amplification using specific primers. Lastly, the developed protocol was evaluated for the detection and quantification of Cryptosporidium parvum in wastewater from different regions in South Africa. The conventional phenol-chloroform extraction method yielded the highest DNA concentration of 223 (±0.71) ng/μl and detected the highest number of Cryptosporidium parvum (1807 (±0.30) copies/ddPCR reaction) compared to other methods evaluated in this study. Additionally, the phenol-chloroform method demonstrated high sensitivity in extracting DNA from as few as one cyst/L of Cryptosporidium parvum, corresponding to 5.93 copies/ddPCR reaction. It was also observed that analysis of both the filtered supernatant and pellets after centrifugation improves the recovery efficiency of oocysts from wastewater by 10.5%, resulting in a total recovery of 64.1%. This optimized protocol was successfully applied to measure protozoan concentration in wastewater from different regions in South Africa. The improved DNA extraction and quantification method proposed in this study would be effective in monitoring protozoan concentration in the environment, which will help in instituting mitigation measures to reduce water-borne infections. [Display omitted] • Droplet digital PCR assay was optimized for detection and quantification of C. parvum from wastewater. • The ddPCR limit of detection for C. parvum was estimated as 1.32 copies per reaction. • The phenol-chloroform method demonstrated high sensitivity in extracting DNA from as few as 1 cyst/L of Cryptosporidium parvum corresponding to 5,93 copies per ddPCR reaction. • The recovery rate of (oo)cysts from wastewater was improved by sample centrifugation followed by supernatant filtration. [ABSTRACT FROM AUTHOR]

Monitoring of COVID-19 infections within communities via wastewater-based epidemiology could provide a cost-effective alternative to clinical testing. This approach, however, still requires improvement for its efficient application. In this paper, we present the use of wastewater-based epidemiology in monitoring COVID-19 infection dynamics in the KwaZulu-Natal province of South Africa, focusing on four wastewater treatment plants for 14 weeks. The SARS-CoV-2 viral load in influent wastewater was determined using droplet digital PCR, and the number of people infected was estimated using published models as well as using a modified model to improve efficiency. On average, viral loads ranged between 0 and 2.73 × 10 5 copies/100 ml, 0-1.52 × 10 5 copies/100 ml, 3 × 10 4 -7.32 × 10 5 copies/100 ml and 1.55 × 10 4 -4.12 × 10 5 copies/100 ml in the four wastewater treatment plants studied. The peak in viral load corresponded to the reported COVID-19 infections within the districts where these catchments are located. In addition, we also observed that easing of lockdown restrictions by authorities corresponded with an increase in viral load in the untreated wastewater. Estimation of infection numbers based on the viral load showed that a higher number of people could potentially be infected, compared to the number of cases reported based on clinical testing. The findings reported in this paper contribute to the field of wastewater-based epidemiology for COVID-19 surveillance, whilst highlighting some of the challenges associated with this approach, especially in developing countries., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

To ensure the safe discharge of treated wastewater to the environment, continuous efforts are vital to enhance the modelling accuracy of wastewater treatment plants (WWTPs) through utilizing state-of-art techniques and algorithms. The integration of metaheuristic modern optimization algorithms that are natlurally inspired with the Fussy Inference Systems (FIS) to improve the modelling performance is a promising and mathematically suitable approach. This study integrates four population-based algorithms, namely: Particle swarm optimization (PSO), Genetic algorithm (GA), Hybrid GA-PSO, and Mutating invasive weed optimization (M-IWO) with FIS system. A full-scale WWTP in South Africa (SA) was selected to assess the validity of the proposed algorithms, where six wastewater effluent parameters were modeled, i.e., Alkalinity (ALK), Sulphate (SLP), Phosphate (PHS), Total Kjeldahl Nitrogen (TKN), Total Suspended Solids (TSS), and Chemical Oxygen Demand (COD). The results from this study showed that the hybrid PSO-GA algorithm outperforms the PSO and GA algorithms when used individually, in modelling all wastewater effluent parameters. PSO performed better for SLP and TKN compared to GA, while the M-IWO algorithm failed to provide an acceptable modelling convergence for all the studied parameters. However, three out of four algorithms applied in this study proven beneficial to be optimized in enhancing the modelling accuracy of wastewater quality parameters., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The release of emerging contaminants (ECs) to the environment is a serious concern due to its health implications on humans, aquatic species, and the development of anti-microbial resistance. This review focuses on the critical analysis of available literature on the prevalence of ECs in the aquatic environment and their removal from wastewater treatment plants (WWTPs) in South Africa. Besides, a risk assessment is performed on the reported ECs from the South African surface water to augment the knowledge towards mitigation of EC pollution, and prioritisation of ECs to assist future monitoring plans and regulation framework. A zone wise classification approach was carried out to identify the spatial inferences and data deficiencies that revealed a non-uniformity in the monitoring of ECs throughout South Africa, with few zones rendering no data. The overarching data mining further revealed that unmanaged urine diverted toilets could be a potential source of EC pollution to groundwater in South Africa. Based on the available literature, it can be deduced that the complete adoption of EC management practices from developed countries might only contribute partly in the mitigation of EC pollution in South Africa. Therefore, an EC monitoring programme specific to the country is recommended which should be based on their occurrence levels, sources and removal in WWTPs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The quality of surface water could be influenced by both anthropogenic and natural factors. This study was designed to determine the impact of informal settlement and wastewater treatment plants on helminth egg contamination of urban rivers and the risks associated with everyday use. We also ascertained the accumulation of these eggs in the river sediments. The study was carried out in two rivers in the eThekwini Municipality of South Africa. Grab samples were taken at different points over a 10-month period. Ascaris spp., hookworm, Toxocara spp., Trichuris spp. and Taenia spp. were the helminth eggs detected in both the water column and sediments, with mean Ascaris spp. eggs of 0-6.3 (± 5.1)/L in the water and 0-6.8 (± 5.2)/kg in sediment samples. The helminth egg concentrations showed seasonal variation, probably due to changes in infection levels of the populations or natural factors, such as rainfall. The informal settlements had a greater impact than treated wastewater. For every 10,000 recreational users of the rivers 19 to 58 may be infected under undisturbed conditions, increasing to 29-88 individuals when the riverbed is disturbed. The risk from agricultural use of the rivers was above the tolerable risk values applicable for wastewater reuse, recommended by the World Health Organization. This calls for a re-evaluation of the policies governing surface water quality assessment, where the inclusion of helminth eggs and sediment monitoring will be critical.

Phenol, a common constituent in many industrial wastewaters is a major pollutant and has several adverse effects on the environment. The potential of various microorganisms to utilize phenol for their metabolic activity has been observed to be an effective means of remediating this toxic compound from the environment particularly wastewater. Five indigenous bacterial isolates (PD1-PD5) were obtained from phenol bearing industrial wastewater using the mineral salts medium. The isolates were further characterized based on their morphology, biochemical reactions and 16S rRNA phylogeny. The 16S rRNA sequence analysis using universal primers (27f/1492r) revealed that PD1, PD2, PD3 and PD4 were closely related to the actinomycete Rhodococcus pyrinidivorans (99%) and PD5 to Pseudomonas aeruginosa (99%). Growth kinetic patterns and phenol degradation abilities of the two representative isolates (PD1 and PD5) were also evaluated. Both the species were effective in utilizing phenol as the sole carbon source and could tolerate phenol concentrations of up to 500 to 600 mg/L. The ability of these isolates to utilize higher concentrations of phenol as their sole carbon source makes them potential candidates and better competitors in the bioremediation process.

South Africa has a rich microalgal biodiversity which has the potential to be used for renewable bio-fuel production in the region. Bioprospecting for oleaginous microalgae in KwaZulu Natal Province, South Africa, resulted in the establishment of a microalgal culture collection system for alternative energy research in the country. A potential hyper-lipid-producing Chlorella spp. strain was isolated, purified, and cultured in supplemented post-chlorinated wastewater for biomass and lipid production at the laboratory scale under batch mode. The microalgal strain was cultivated in different strengths of BG-11 media supplemented with wastewater from a local municipal domestic wastewater treatment plant. The Chlorella spp. was grown using ambient dissolved carbon dioxide in shake flasks under photosynthetically active radiation (±120 μmol m⁻² s⁻¹). Microalgal biomass and lipid productivity were monitored at 24-h intervals in the batch mode. The microalgal biomass was analyzed by direct light microscopy and indirectly by spectrophotometry at 600 nm, and the lipids were extracted and quantified. The growth rate of the Chlorella spp. was enhanced in post-chlorinated wastewater supplemented with 5 mM NaNO₃ with maximal biomass productivity. A dramatic increase in lipid yield was achieved with the post-chlorinated wastewater supplemented with 25 mM NaNO₃. Low dosages of free chlorine were found to enhance microalgal growth. These findings serve as a basis for further scale-up trials using municipal wastewater as a medium for microalgal biomass and lipid production.

This WRC funded project has studied the appropriateness of the ABR (anaerobic baffled reactor) for on-site primary sanitation in low-income communities. A 3,000 L pilot reactor was located at the Kingsburgh wastewater treatment plant south of Durban, South Africa. Feed to the reactor was raw domestic wastewater containing a significant proportion of particulate organic matter. The compartments of the ABR were routinely monitored for pH, COD, and gas production, among other physical-chemical determinants. The microbial population in each compartment was analysed by fluorescent in situ hybridisation, using general oligonucleotide probes for eubacteria and archeae and a suite of 10 genera or family specific probes. Scanning electron microscopy was conducted on the sludge fraction of each compartment. Mixed fractions from each compartment were also analysed for health-related indicator bacteria (total coliforms and E. coli). Results indicated that methanogenesis was not occurring to the expected extent in the latter compartments, and that this was probably due to a hydraulic load limitation. This contrasted with earlier studies on industrial effluent, for which the organic load was exclusively in soluble form. Inactivation of health-related indicator bacteria was less than 1 log, indicating the need for an additional post-treatment of the effluent to protect community health.