The role of effect-based methods to address water quality monitoring in South Africa: a developing country's struggle.

Water is an important resource, and it is a worldwide struggle to provide water of good quality to the whole population. Despite good governing laws and guidelines set in place to help protect the water resources and ensure it is of good quality for various consumers, the water quality in South Africa is worsening due to lack of management. The deteriorating infrastructure is becoming progressively worse, due to corruption and insufficient funds. The ever-increasing number of toxicants, as well as the identification of emerging chemicals of concern, are also challenges South Africa is facing. Chemical analysis cannot determine the total biological effect of a mixture of chemical compounds, but this shortcoming can be addressed by adding effect-based methods (EBMs) to water quality monitoring programmes. In this paper, the current status of water quality monitoring in South Africa is discussed, as well as the capacity of the country to add EBMs to its water quality monitoring programmes to protect and improve human and animal life. Created in Biorender.com.

Data on UPLC/MS method validation for the biodegradation of pharmaceuticals and intermediates by a fungal consortium and on T47DK-Bluc reporter gene assay to assess the reduction of their estrogenic activity.

In term of pharmaceutical and their intermediate compounds analysis, UPLC/MS method is a valuable equipment to achieve better confirmation on their biodegradation by fungi. The T47D-KBluc reporter gene assay is an appropriate tool to investigate to removal of estrogenic and antiestrogenic activities of pharmaceuticals and their metabolites from a synthetic wastewater. A consortium of isolated South African indigenous fungi Aspergillus niger, Mucor circinelloides, Trichoderma longibrachiatum, Trametes polyzona and Rhizopus microspores was found to perform a removal of pharmaceuticals and their metabolites and to reduce their estrogenic activity below the limit of detection in a sequencing batch reactor. Here are presented data regarding the phenolic compounds list and the method validation for UPLC/MS analysis used for selected pharmaceutical compounds namely carbamazepine, diclofenac, ibuprofen and their metabolites, as well as the T47D-KBluc bioassay using as positive control, the agonist E2 for estrogenic activity and the antagonist ICI 182,780 for antiestrogenic activity. For better understanding of the data presented in this paper, please see the research paper "Removal of pharmaceutical' estrogenic activity of sequencing batch reactor effluents assessed in the T47DK-Bluc reporter gene assay" [1].

Removal of pharmaceutical' estrogenic activity of sequencing batch reactor effluents assessed in the T47D-KBluc reporter gene assay.

Various water treatment processes may be ineffective to remove pharmaceutical compounds (PhCs) and their by-products, leading to endocrine-disruptive activity that might be detrimental to wildlife and human health. This study investigated the degradation of carbamazepine (CBZ), diclofenac (DCF), ibuprofen (IBP), and their intermediates, as well as estrogenic activity that is not effectively removed by conventional methods. A consortium of isolated South African indigenous fungi A. niger, M. circinelloides, T. polyzona, T. longibrachiatum and R. microsporus, was used in a sequencing batch reactor (SBR) to remove PhCs, their intermediates and strongly reduce their estrogenic activity. The fungal ligninolytic enzymatic activity was determined for laccase (Lac), manganese peroxidase (MnP) and lignin peroxidase (LiP) using a spectrophotometric method. The biodegradation of PhCs and their intermediates was monitored by SPE-UPLC/MS. The in vitro estrogenic activity was assessed in the T47D-KBluc reporter gene assay. Lac, MnP and LiP production appeared to be biomass growth dependent. During a lag phase of growth, a constant biomass of about 122.04 mg/100 mL was recorded with average enzymatic activity around 63.62 U/L for Lac, 151.91 U/L for MnP and 42.12 U/L for LiP. The exponential growth phase from day 7 to day 17, was characterised by a biomass increase of 124.46 units, and an increase in enzymatic activity of 9.91 units for Lac, 99.03 units for MnP and 44.24 units for LiP. These enzymes played an important synergistic role in PhCs degradation in the cytochrome P450 system. A decrease of 13.89%, 29.7% and 16.15% in PhC concentrations was observed for CBZ, DCF and IBP, respectively, and their intermediates were identified within 4 h of incubation. The removal efficiency achieved after 24 h in the SBR was about 89.77%, 95.8% and 91.41% for CBZ, DCF and IBP, respectively. The estradiol equivalent (EEq) values of 1.71 ±â€¯0.30 ng/L and 2.69 ±â€¯0.17 ng/L were recorded at the start-up time and after 4 h, respectively. The presence of intermediates was found to induce estrogenic activity. The EEq values after 24 h incubation was found to be below the LoQ and below the LoD of the assay. None of the samples exhibited any anti-estrogenic activity. The fungal consortium inoculum was found to induce toxicity at a 0.4× concentration, as observed under a microscope. This study revealed that the use of the fungal consortium can remove the estrogenic activity of pharmaceutical metabolites, which appeared to be the most significant contributors to the endocrine-disrupting activity of the wastewater treatment plant effluents.

In vitro effects of 2-methoxyestradiol on MCF-12A and MCF-7 cell growth, morphology and mitotic spindle formation.

The influence of 2-methoxyestradiol (2ME) was investigated on cell growth, morphology and spindle formation in a tumorigenic (MCF-7) and non-tumorigenic (MCF-12A) epithelial breast cell line. Inhibition of cell growth was more pronounced in the MCF-7 cells compared to the MCF-12A cells following 2ME treatment. Dose-dependent studies (10(-5)-10(-9) M) revealed that 10(-6) M 2ME inhibited cell growth by 44% in MCF-12A cells and by 84% in MCF-7 cells (p-value < 0.05). 2ME-treated MCF-7 cells showed abnormal metaphase cells, membrane blebbing, apoptotic cells and disrupted spindle formation. These observations were either absent or less prominent in MCF-12A cells. 2ME had no effect on the length of the cell cycle between S-phase and the time a mitotic peak was reached in either cell line but MCF-7 cells were blocked in mitosis with no statistically significant alterations in the phosphorylation status of Cdc25C. Nevertheless, Cdc2 activity was significantly increased in MCF-7 cells compared to MCF-12A cells (p-value < 0.05). The results indicate that 2ME disrupts mitotic spindle formation and enhances Cdc2 kinase activity, leading to persistence of the spindle checkpoint and thus prolonged metaphase arrest that may result in the induction of apoptosis. The tumorigenic MCF-7 cells were especially sensitive to 2ME treatment compared to the normal MCF-12A cells. Therefore, differential mechanism(s) of growth inhibition are evident between the normal and tumorigenic cells.