Influence of seasonality, wastewater treatment plant process, geographical location and environmental parameters on bacterial community selection in activated sludge wastewater treatment plants treating municipal sewage in South Africa.

This is the first comprehensive study that focusses on the correlation between the bacterial community composition and a range of previously identified selective criteria in activated sludge wastewater treatment plants on the African continent. Multivariate statistical analyses were used to determine the relative significance of the geographical location (factor: site), wastewater treatment plant process (factor: configuration), seasonality (factor: season), and environmental parameters on the bacterial communities in nine wastewater treatments plants from two sites in South Africa using terminal restriction fragment length polymorphism as a screening tool to rationalize the number of samples (to 50 samples) for high throughput (Illumina MiSeq) sequencing. Site was the most significant factor (Global ANOSIM R value = 0.91, p = 0.001), and it was established that the inter-site differences were not climatic in origin but related to differences in the composition of the influent and activated sludge. Previous studies that have reported associations between microbial community structure and environmental parameters have measured influent chemistry, and this is the first time, to our knowledge, that the comprehensive chemical character of activated sludge itself has been included in this type of study. It was found using BEST analysis that the activated sludge ammonia, activated sludge total phosphate and influent chemical oxygen demand were the most significant (p < 0.001) drivers for inter-site bacterial community selection (ANOSIM Global R values of 0.862, 0.782 and 0.428, respectively). This link would not have been established with only influent chemical analyses as there was no significant difference (t-test, p > 0.05) in the average influent phosphate concentrations between the 2 sites, but there was a highly significant difference (p < 0.001, t (15.5)>t-crit (2.01)) in the activated sludge total phosphate concentrations (20.8 ± 17.0 and 127.8 ± 40.2 mg/L). This is notable for all future studies on a global level aimed at identifying factors for selection of microbial communities in activated sludge.

Bacterial nitrogen fixation in sand bioreactors treating winery wastewater with a high carbon to nitrogen ratio.

Heterotrophic bacteria proliferate in organic-rich environments and systems containing sufficient essential nutrients. Nitrogen, phosphorus and potassium are the nutrients required in the highest concentrations. The ratio of carbon to nitrogen is an important consideration for wastewater bioremediation because insufficient nitrogen may result in decreased treatment efficiency. It has been shown that during the treatment of effluent from the pulp and paper industry, bacterial nitrogen fixation can supplement the nitrogen requirements of suspended growth systems. This study was conducted using physicochemical analyses and culture-dependent and -independent techniques to ascertain whether nitrogen-fixing bacteria were selected in biological sand filters used to treat synthetic winery wastewater with a high carbon to nitrogen ratio (193:1). The systems performed well, with the influent COD of 1351 mg/L being reduced by 84-89%. It was shown that the nitrogen fixing bacterial population was influenced by the presence of synthetic winery effluent in the surface layers of the biological sand filters, but not in the deeper layers. It was hypothesised that this was due to the greater availability of atmospheric nitrogen at the surface. The numbers of culture-able nitrogen-fixing bacteria, including presumptive Azotobacter spp. exhibited 1-2 log increases at the surface. The results of this study confirm that nitrogen fixation is an important mechanism to be considered during treatment of high carbon to nitrogen wastewater. If biological treatment systems can be operated to stimulate this phenomenon, it may obviate the need for nitrogen addition.

Selection of diazotrophic bacterial communities in biological sand filter mesocosms used for the treatment of phenolic-laden wastewater.

Agri effluents such as winery or olive mill wastewaters are characterized by high phenolic concentrations. These compounds are highly toxic and generally refractory to biodegradation. Biological sand filters (BSFs) represent inexpensive, environmentally friendly, and sustainable wastewater treatment systems which rely vastly on microbial catabolic processes. Using denaturing gradient gel electrophoresis and terminal-restriction fragment length polymorphism, this study aimed to assess the impact of increasing concentrations of synthetic phenolic-rich wastewater, ranging from 96 mg L(-1) gallic acid and 138 mg L(-1) vanillin (i.e., a total chemical oxygen demand (COD) of 234 mg L(-1)) to 2,400 mg L(-1) gallic acid and 3,442 mg L(-1) vanillin (5,842 mg COD L(-1)), on bacterial communities and the specific functional diazotrophic community from BSF mesocosms. This amendment procedure instigated efficient BSF phenolic removal, significant modifications of the bacterial communities, and notably led to the selection of a phenolic-resistant and less diverse diazotrophic community. This suggests that bioavailable N is crucial in the functioning of biological treatment processes involving microbial communities, and thus that functional alterations in the bacterial communities in BSFs ensure provision of sufficient bioavailable nitrogen for the degradation of wastewater with a high C/N ratio.

Ammonium oxidizing bacterial populations in South African activated sludge wastewater treatment plants.

This is the first study that describes ammonium oxidizing bacterial populations and correlations of these populations with a range of criteria in activated sludge wastewater treatment plants in South Africa. In this study, not only the influent but also the activated sludge chemistry was comprehensively characterized. Multivariate statistical analyses were used to determine the relative significances of the geographical location (factor: site), wastewater treatment plant process (factor: configuration), seasonality (factor: season), and environmental parameters on the ammonium oxidizing bacterial genera in six municipal activated sludge wastewater treatments plants from two sites (the cities of Cape Town and Ekurhuleni). The geographical location (site) was significant for selection of the ammonium oxidizing genera (Global ANOSIM R value = 0.538, p = 0.001). It was established that the inter-site differences were not climatic in origin, nor related to the composition of the influent, but were rather driven by the activated sludge chemistry. It was found using BEST analysis that the activated sludge ammonia, activated sludge total phosphate, and activated sludge total chemical oxygen demand were the most significant (p < 0.001) drivers for ammonium oxidizing bacterial selection (ANOSIM Global R value 0.419) and were significantly higher in the activated sludge from the City of Cape Town wastewater treatment plants. Nitrosospira was the most abundant ammonium oxidizing bacterial genus, with notably higher relative and estimated actual abundances in the City of Cape Town wastewater treatment plants than the City of Ekurhuleni wastewater treatment plants. The strong selection of Nitrosospira in the City of Cape Town wastewater treatment plants with higher nutrient concentrations strongly suggests that high concentrations of activated sludge ammonia, activated sludge total phosphate, and activated sludge total chemical oxygen demand are key selective drivers for this genus. PRACTITIONER POINTS: First comprehensive study describing ammonium oxidizing bacterial populations in Southern African domestic activated sludge wastewater treatment plants. The geographical location (site) was significant for selection of different ammonium oxidizing genera (Global ANOSIM R value = 0.538, p = 0.001). Inter-site differences driven by the activated sludge chemistry, not climate or influent wastewater composition. Selection of Nitrosospira driven by high concentrations of activated sludge ammonia, total phosphate and total chemical oxygen demand.

Treatment wetlands and phyto-technologies for remediation of winery effluent: Challenges and opportunities.

The composition and concentration of contaminants present in winery wastewater fluctuate through space and time, presenting a challenge for traditional remediation methods. Bio-hydrogeochemical engineered systems, such as treatment wetlands, have been demonstrated to effectively reduce contaminant loads prior to disposal or reuse of the effluent. This review identifies and details the status quo and challenges associated with (i) the characteristics of winery wastewater, and the (ii) functional components, (iii) operational parameters, and (iv) performance of treatment wetlands for remediation of winery effluent. Potential solutions to challenges associated with these aspects are presented, based on the latest literature. A particular emphasis has been placed on the phytoremediation of winery wastewater, and the rationale for selection of plant species for niche bioremediatory roles. This is attributed to previously reported low-to-negative removal percentages of persistent contaminants, such as salts and heavy metals that may be present in winery wastewater. A case for the inclusion of selected terrestrial halophytes in treatment wetlands and in areas irrigated using winery effluent is discussed. These are plant species that have an elevated ability to accumulate, cross-tolerate and potentially remove a range of persistent contaminants from winery effluent via various phytotechnologies (e.g., phytodesalination).

Functional Microbial Communities in Hybrid Linear Flow Channel Reactors for Desulfurization of Tannery Effluent.

Recent research has demonstrated that hybrid linear flow channel reactors (HLFCRs) can desulfurize tannery effluent via sulfate reduction and concurrent oxidation of sulfide to elemental sulfur. The reactors can be used to pre-treat tannery effluent to improve the efficiency of downstream anaerobic digestion and recover sulfur. This study was conducted to gain insight into the bacterial communities in HLFCRs operated in series and identify structure-function relationships. This was accomplished by interpreting the results obtained from amplicon sequencing of the 16S rRNA gene and quantification of the dissimilatory sulfite reducing (dsrB) gene. In an effort to provide a suitable inoculum, microbial consortia were harvested from saline estuaries and enriched. However, it was found that bioaugmentation was not necessary because native communities from tannery wastewater were selected over exogenous communities from the enriched consortia. Overall, Dethiosulfovibrio sp. and Petrimonas sp. were strongly selected (maximum relative abundances of 29% and 26%, respectively), while Desulfobacterium autotrophicum (57%), and Desulfobacter halotolerans (27%) dominated the sulfate reducing bacteria. The presence of elemental sulfur reducing genera such as Dethiosulfovibrio and Petrimonas is not desirable in HLFCRs, and strategies to counter their selection need to be considered to ensure efficiency of these systems for pre-treatment of tannery effluent.

Impact of Physicochemical Parameters on the Diversity and Distribution of Microbial Communities Associated with Three South African Peatlands.

Peatlands are complex wetland-like ecosystems that harbor diverse microbial communities. In this study, the microbial communities (fungal and actinobacterial) associated with an unimpacted peatland (Vankervelsvlei; VV), an impacted peatland (Goukou River system; GK), and a developing peatland (Nuwejaars River system; NR) were determined through ITS and 16S rRNA metataxonomic analyses. Unidentified Acidimicrobiales dominated in GK and NR, unidentified Intrasporangiaceae and Solirubobacterales in NR, and Corynebacterium, Propionibacterium, and Streptomyces species in VV. The fungal phyla, Ascomycota and Basidiomycota, dominated all three sites, and harbored unique fungal taxa belonging to a wide range of fungal guilds. Physicochemical properties of the peat collected from the three sites were analyzed in association with microbial community structures in order to determine which parameters acted as the main drivers for microbial diversity. BEST analysis (linking microbial diversity patterns to environmental variables) showed that nitrogen (N), aluminum (Al), phosphorus (P), and potassium (K) were the most significant physicochemical drivers of actinobacterial community structure, while iron (Fe) and humification were the environmental parameters that affected the fungal communities the most. In conclusion, this study has provided some insight into the fungal and actinobacterial communities associated with three South African peatlands and the main environmental drivers that influence these communities.

The effect of biogenic and chemically manufactured silver nanoparticles on the benthic bacterial communities in river sediments.

This study was conducted to determine and compare the effect of chemically-synthesised and biogenic silver nanoparticles on the benthic bacterial community structure in mesocosms containing sediment from three rivers in geographical sites with different population densities (low, medium, high), and therefore likely to be associated with respective low, moderate and high degrees of anthropogenic input. The nanoparticles were applied at the upper limit expected to accumulate in impacted environments (4 µg kgsed-1). The biomass, concentrations of elements, including selection metals (P, K, Na, K, Ca, Mg, Zn, Cu, Al, Ag) were all significantly higher at the high density than at the low density sites. Bacterial community profiling (terminal restriction fragment length polymorphism and amplicon sequencing) showed that the bacterial community structure in the sediments from the high population density site were resilient to environmental perturbations [adjustment from in-situ to ex-situ (laboratory) conditions], as well as to exposure to silver nanoparticles, with the converse being true for the low population density site. Results obtained from amplicon sequencing were interrogated to the lowest taxonomic level with a relative abundance >5%. Proteobacteria was the most abundant phylum in all the sediments. Notable resistance (increased relative abundance) to one or both forms of silver nanoparticles was seen in the class Thermoleophilia, and the orders Myxococcales, Bacteriodales, Pirellules CCU21 and iii 1-15 (class Acidobacteria 6). Conversely, sensitivity was demonstrated in the family Koribacteraceae and the orders Rhizobiales, Ellin 329 and Gemmatales. It is recommended that pro-active environmental monitoring is performed in aquatic systems receiving point source pollution from wastewater treatment plants in order to assess the accumulation of silver nanoparticles. If necessary, measures should be implemented to mitigate the entry of silver nanoparticles, especially into more vulnerable environments.

Selection of Clostridium spp. in biological sand filters neutralizing synthetic acid mine drainage.

In this study, three biological sand filter (BSF) were contaminated with a synthetic iron- [1500 mg L⁻¹ Fe(II), 500 mg L⁻¹ Fe(III)] and sulphate-rich (6000 mg L⁻¹ SO4²â») acid mine drainage (AMD) (pH = 2), for 24 days, to assess the remediation capacity and the evolution of autochthonous bacterial communities (monitored by T-RFLP and 16S rRNA gene clone libraries). To stimulate BSF bioremediation involving sulphate-reducing bacteria, a readily degradable carbon source (glucose, 8000 mg L⁻¹) was incorporated into the influent AMD. Complete neutralization and average removal efficiencies of 81.5 (±5.6)%, 95.8 (±1.2)% and 32.8 (±14.0)% for Fe(II), Fe(III) and sulphate were observed, respectively. Our results suggest that microbial iron reduction and sulphate reduction associated with iron precipitation were the main processes contributing to AMD neutralization. The effect of AMD on BSF sediment bacterial communities was highly reproducible. There was a decrease in diversity, and notably a single dominant operational taxonomic unit (OTU), closely related to Clostridium beijerinckii, which represented up to 65% of the total community at the end of the study period.

Microbial community structure stability, a key parameter in monitoring the development of constructed wetland mesocosms during start-up.

Constructed wetlands (CWs) are known to be effective for treating waste streams, and pilot-scale CWs are useful for assessing the impact of pollutants and their remediation. However, little is known with respect to the establishment of these mesocosm systems or the parameters which should be monitored in assessing system equilibration, i.e. when they present stabilised physical and biological patterns. The aim of this study was to evaluate the temporal aspects of CW equilibration as a basis for future studies of system response to amendment. Microbial biomass and hydraulic conductivity values were monitored and microbial community fingerprints were obtained using denaturing gradient gel electrophoresis (DGGE). This study showed that microbial community fingerprinting provides a valuable tool for assessing the time scales of equilibration, as it was the last parameter which stabilised during the equilibration period. Hydraulic conductivity was also an important parameter in determining the time scale for initiation of the equilibration process during the study. For a CW of the dimensions used (173 cm long/106 cm large/30 cm depth), community equilibration times demonstrated on the basis of similar microbial community structures were found to be on the order of 100 days.