Review: Pharmaceutical and personal care products (PPCPs) as endocrine disrupting contaminants (EDCs) in South African surface waters

Globally, water resources are under constant threat of being polluted by a diverse range of man-made chemicals, and South Africa is no exception. These  contaminants can have detrimental effects on both human and wildlife health. It is increasingly evident that several chemicals may modulate endocrine system pathways in vertebrate species, and these are collectively referred to as endocrine disrupting contaminants (EDCs). Although the endocrine-disrupting effect of water pollutants has been mainly linked to agricultural pesticides and industrial effluents, other pollutants such as pharmaceuticals and personal care products (PPCPs) are largely unnoticed, but also pose a potentially significant threat. Here we present for the first time in a South African context, a summarised list of PPCPs and other EDCs detected to date within South African water systems, as well as their possible endocrine-disrupting effect in-vitro and in-vivo. This review addresses other factors which should be investigated in future studies, including endocrine disruption, PPCP metabolites, environmental toxicology, and antibiotic resistance. The challenges of removing EDCs and other pollutants at South African wastewater treatment works (WWTWs) are also highlighted. The need for focused research involving both in-vitro and in-vivo studies to detect PPCPs in water systems, and to delineate adverse outcome pathways (AOPs) of priority PPCPs to aid in environmental impact  assessment (EIA), are discussed.Keywords: pharmaceutical, endocrine disruption, wastewater, sewage wate

The fate of pharmaceuticals and personal care products (PPCPs), endocrine disrupting contaminants (EDCs), metabolites and illicit drugs in a WWTW and environmental waters.

A large number of emerging contaminants (ECs) are known to persist in surface waters, and create pressure on wastewater treatment works (WWTW) for their effective removal. Although a large database for the levels of these pollutants in water systems exist globally, there is still a lack in the correlation of the levels of these pollutants with possible long-term adverse health effects in wildlife and humans, such as endocrine disruption. The current study detected a total of 55 ECs in WWTW influent surface water, 41 ECs in effluent, and 40 ECs in environmental waters located upstream and downstream of the plant. A list of ECs persisted through the WWTW process, with 28% of all detected ECs removed by less than 50%, and 18% of all ECs were removed by less than 25%. Negative mass balances of some pharmaceuticals and metabolites were observed within the WWTW, suggesting possible back-transformation of ECs during wastewater treatment. Three parental illicit drug compounds were detected within the influent of the WWTW, with concentrations ranging between 27.6 and 147.0 ng L−1 for cocaine, 35.6–120.6 ng L−1 for mephedrone, and 270.9–450.2 ng L−1 for methamphetamine. The related environmental risks are also discussed for some ECs, with particular reference to their ability to disrupt endocrine systems. The current study propose the potential of the pharmaceuticals carbamazepine, naproxen, diclofenac and ibuprofen to be regarded as priority ECs for environmental monitoring due to their regular detection and persistence in environmental waters and their possible contribution towards adverse health effects in humans and wildlife

Identification of synergistic interactions among microorganisms in biofilms by digital image analysis

Digital image analysis showed that reductions in biofilm plating efficiency were due to the loss of protection provided by two benzoate-degrading strains of Pseudomonas fluorescens. This loss in protection was due to the spatial separation of the protective organisms from benzoate-sensitive organisms during the dilution process. Communities were cultivated in flow cells irrigated with trypticase soy broth. When the effluent from these flow cells was plated on 0.15% benzoic acid, satellite colonies formed only in the vicinity of primary colonies. A digital image analysis procedure was developed to measure the size and spatial distribution of these satellites as a function of distance from the primary colony. The size of satellites served as a measure of growth, and the number per unit area served as a measure of survival. At the three dilutions tested, the size and concentration of satellite colonies varied inversely with distance from the primary colonies. When these measurements were plotted, the slopes were used to quantify the effect of bacterial association on the growth and survivability of the satellites. In the absence of the primary colonies, satellites grew in axenic culture only at low benzoate concentrations. Thus benzoate-degrading organisms are capable of creating a protective microenvironment for other members of biofilm communities

Form and Function of Clostridium Thermocellum Biofilms

The importance of bacterial adherence has been acknowledged in microbial lignocellulose conversion studies; however, few reports have described the function and structure of biofilms supported by cellulosic substrates. We investigated the organization, dynamic formation, and carbon flow associated with biofilms of the obligately anaerobic cellulolytic bacterium Clostridium thermocellum 27405. Using noninvasive, in situ fluorescence imaging, we showed biofilms capable of near complete substrate conversion with a characteristic monolayered cell structure without an extracellular polymeric matrix typically seen in biofilms. Cell division at the interface and terminal endospores appeared throughout all stages of biofilm growth. Using continuous-flow reactors with a rate of dilution (2 h1) 12-fold higher than the bacterium’s maximum growth rate, we compared biofilm activity under low (44 g/liter) and high (202 g/liter) initial cellulose loading. The average hydrolysis rate was over 3-fold higher in the latter case, while the proportions of oligomeric cellulose hydrolysis products lost from the biofilm were 13.7% and 29.1% of the total substrate carbon hydrolyzed, respectively. Fermentative catabolism was comparable between the two cellulose loadings, with ca. 4% of metabolized sugar carbon being utilized for cell production, while 75.4% and 66.7% of the two cellulose loadings, respectively, were converted to primary carbon metabolites (ethanol, acetic acid, lactic acid, carbon dioxide). However, there was a notable difference in the ethanol-to-acetic acid ratio (g/g), measured to be 0.91 for the low cellulose loading and 0.41 for the high cellulose loading. The results suggest that substrate availability for cell attachment rather than biofilm colonization rates govern the efficiency of cellulose conversion

Live-streaming: time-lapse video evidence of novel streamer formation mechanism and varying viscosity

Time-lapse videos of growing biofilms were analyzed using a background subtraction method, which removed camouflaging effects from the heterogeneous field of view to reveal evidence of streamer formation from optically dense biofilm segments. In addition, quantitative measurements of biofilm velocity and optical density, combined with mathematical modeling, demonstrated that streamer formation occurred from mature, high-viscosity biofilms. We propose a streamer formation mechanism by sudden partial detachment, as opposed to continuous elongation as observed in other microfluidic studies. Additionally, streamer formation occurred in straight microchannels, as opposed to serpentine or pseudo-porous channels, as previously reported

The assessment of phytoplankton dynamics in two reservoirs in southern Africa with special reference to water abstraction for inter-basin transfers and potable water production

Toxic phytoplankton in the aquatic ecosystems are dynamic, affecting water quality. It remains unclear as to how possible toxic phytoplankton assemblages vary vertically and temporally in Swakoppoort and Von Bach dams, located in a dry subtropical desert region in central Namibia. The following variables were analyzed: pH, Secchi depths, turbidity, water temperature, total phosphorus, orthophosphate, chlorophyll-a, phytoplankton cells, and water depths. Cyanobacteria dominated the phytoplankton community in the autumn, winter and spring (dry) and summer (wet) seasons, at all the depth ranges in both dams. Microcystis dominated the vertical and temporal dynamics, followed by Dolichospermum. In the dry seasons, higher cyanobacteria cell numbers were observed in comparison to the rainy season in both dams. Spring blooms of cyanobacteria were evident in the Von Bach Dam while autumn and spring cyanobacteria blooms were observed in the Swakoppoort Dam. In the Swakoppoort Dam, the preferable depth ranges for toxic cyanobacteria species were at 5 to 10 m while in the Von Bach Dam at 0 to 5 m range. The findings of the current study indicate that the traditional selective withdrawal of water in the two dams should be performed with vertical and temporal dynamics of possible toxic cyanobacteria accounted for to aid the abstraction of water with the lowest possible toxic phytoplankton numbers, which could lower the public health risk.http://www.mdpi.com/journal/waterParaclinical Science

A flow cell simulating a subsurface rock fracture for investigations of groundwater-derived biofilms

Laboratory scale continuous-flow-through chambers (flow cells) facilitate the observation of microbes in a controlled, fully hydrated environment, although these systems often do not simulate the environmental conditions under which microorganisms are found. We developed a flow cell that mimics a subsurface groundwater-saturated rock fracture and isamenable to confocal laser scanning microscopy while allowing for the simple removal of the attached biomass. This flow cell was used to investigate the effect of toluene, a representative contaminant for non-aqueous phase liquids, on groundwater-derived biofilms. Reduced average biofilm biomass and thickness, and diminished diversity of amplifiable 16S rRNA sequences were observed for biofilms that developed in the presence of toluene, compared to the biofilms grown in the absence of toluene. The flow cell also allowed the detection of fluorescent protein-labelled cells

Accumulation and degradation of diclofop methyl by cultured biofilm communities

Traditionally, bacteria are viewed as unicellular organisms and are studied as isolated cell lines. However, in natural systems they rarely exist as pure cultures, and thus there is a need to study them as communities under defined laboratory conditions. In this study, a degradative microbial community consisting of nine bacterial and one algal species was isolated from soil using the herbicide diclofop methyl as sole carbon source. The presence of the algae, or addition of an exogenous carbon source, significantly increased diclofop mineralization in continuous flow (36% increase) and batch (11% increase) cultures. Pure cultures isolated from this bacterial consortium could not mineralize diclofop. However, when supplied with an additional carbon source, two strains could degrade more than 25% of it to CO2. Biofilm formation also resulted in more efficient degradation (36% increase). These observations indicated that interspecies interactions and spatial organization within biofilms enhance degradative efficiency. Subsequent analysis of the degradative biofilms, using scanning confocal laser microscopy, revealed distinctive spatial relationships among members of the consortium when grown on diclofop. These relationships, such as the formation of cell clusters within biofilms and an irregular surface topography, were absent when more labile substrates were provided as the carbon source. Stable biofilms having these distinctive features, normally developed in 14 to 21 days. Their development was accompanied by an increase in autofluorescence, indicating accumulation of diclofop and its aromatic breakdown products. Probe mass spectroscopy confirmed the presence of these compounds in the biofilm matrix. Most accumulation occurred in cell capsules and exopolymeric materials within specific regions of the biofilm matrix. Additional evidence for spatial organization in the degradative biofilms was obtained using a set of FITC-conjugated probes. These revealed that regions of the exopolymer matrix which accumulated diclofop had a unique chemical composition and charge. These regions were not present in biofilms grown on a more labile growth medium. Fluorescence extinction and radioisotopic techniques demonstrated that the adsorbed diclofop was utilized during periods of carbon deprivation, indicating that exopolymers can store adsorbed carbon for subsequent utilization

CO2 Production as an Indicator of Biofilm Metabolism▿ †

Biofilms are important in aquatic nutrient cycling and microbial proliferation. In these structures, nutrients like carbon are channeled into the production of extracellular polymeric substances or cell division; both are vital for microbial survival and propagation. The aim of this study was to assess carbon channeling into cellular or noncellular fractions in biofilms. Growing in tubular reactors, biofilms of our model strain Pseudomonas sp. strain CT07 produced cells to the planktonic phase from the early stages of biofilm development, reaching pseudo steady state with a consistent yield of ∼107 cells·cm−2·h−1 within 72 h. Total direct counts and image analysis showed that most of the converted carbon occurred in the noncellular fraction, with the released and sessile cells accounting for <10% and <2% of inflowing carbon, respectively. A CO2 evolution measurement system (CEMS) that monitored CO2 in the gas phase was developed to perform a complete carbon balance across the biofilm. The measurement system was able to determine whole-biofilm CO2 production rates in real time and showed that gaseous CO2 production accounted for 25% of inflowing carbon. In addition, the CEMS made it possible to measure biofilm response to changing environmental conditions; changes in temperature or inflowing carbon concentration were followed by a rapid response in biofilm metabolism and the establishment of new steady-state conditions

Microbes at Surface-Air Interfaces: The Metabolic Harnessing of Relative Humidity,Surface Hygroscopicity and Oligotrophy or Resilience

The human environment is predominantly not aqueous, and microbes are ubiquitous at the surface-air interfaces with which we interact. Yet microbial studies at surface-air interfaces are largely survival-oriented, whilst microbial metabolism has overwhelmingly been investigated from the perspective of liquid saturation. This study explored microbial survival and metabolism under desiccation, particularly the influence of relative humidity, surface hygroscopicity, and nutrient availability on the interchange between these two types of behavior. The combination of a hygroscopic matrix (i.e., clay or 4,000 MW polyethylene glycol) and high relative humidity (RH) resulted in persistent measurable microbial metabolism during desiccation. In contrast, no microbial metabolism was detected at (a) hygroscopic interfaces at low RH, and (b) less hygroscopic interfaces (ie., sand and plastic/glass) at high or low RH. Cell survival was conversely inhibited at high RH and promoted at low RH, irrespective of surface hygroscopicity. Based on this demonstration of metabolic persistence and survival inhibition at high relative humidity, it was proposed that biofilm metabolic rates might inversely influence whole-biofilm resilience, with ‘resilience’ defined in this study as a biofilm’s capacity to recover from desiccation. The concept of whole-biofilm resilience being promoted by oligotrophy was supported in desiccation-tolerant Arthrobacter spp. biofilms, but not in desiccation-sensitive Pseudomonas aeruginosa biofilms. The ability of microbes to interact with surfaces to harness water vapor during desiccation was demonstrated, and potentially to harness oligotrophy (the most ubiquitous natural condition facing microbes) for adaptation to desiccation