Master of Science / Department of Civil Engineering / Alok Bhandari / Antibiotics and pharmaceuticals are used to improve the quality of life worldwide. However, incomplete metabolism in humans has resulted in the release of large amounts of pharmaceutical drugs into municipal wastewater treatment plant. Past research has shown the release of antibiotic resistant organisms through wastewater effluents into streams and several studies have reported the occurrence of antibiotic resistant bacteria in major U.S. Rivers. Antibiotic resistant bacteria evolve and are selected by long-term environmental exposure to the low concentrations of antibiotics at the ng /L to g/L range. Infections caused by antibiotic resistant organisms are difficult to treat. The aim of this study was to analyze antibiotic resistance patterns in selected wastewater bacteria that include fecal coliforms, Escherichia coli and enterococci. Microorganisms in municipal wastewater treatment plant influent, secondary clarifier effluent and disinfected effluent were plated in the presence of predetermined concentrations of selected antibiotics. These antibiotics included ciprofloxacin, sulfamethoxazole/ trimethoprim and vancomycin. The diversity of enterococci was further investigated with PCR analysis. Fecal coliforms, E. coli and enterococci were found to be resistant or highly resistant to one or more target antibiotics in the influent and secondary clarifier (SC) effluent. Biological treatment reduced the number of overall and resistant bacteria in the SC effluent sample. UV disinfection was generally very effective and eliminated all fecal indicator organisms.

A growing population, urbanization and an increase in the number of industries is causing an increase in sewage sludge (SS) that needs to be either beneficially used or disposed of. Application of SS to agricultural lands is a well-known practice but the plant available phosphate and phosphate fertilizer value of SS has been of concern. This is especially due to the wide variety of soils that SS is applied to and the different phosphate removal processes that sewage waste water undergoes at the water care works and the source used to produce SS. Phosphate phyto-availability and phosphate fertilizer value of petrochemical and municipal wastewater sludges (SS) were determined in four different soils using an incubation study over 168 days, a pot trial over 42 days and a field trial over one season. Phosphate phyto-availability was determined/calculated by means of an incubation approach. Soils were incubated with sludge as well as mono ammonium phosphate (MAP). The soils were then subjected to a Bray-1 extraction after a certain time (42 days, 168 days). The relative phosphate fertilizer value (RPFV) was then expressed as a percentage of the Bray-1 extractability of the sludge-amended soil, relative to the MAP-amended soil. The influence of soil properties, especially clay content, and sludge properties namely phosphate extraction method (chemical and biological) from waste water stream, was investigated to determine the effect on phytoavailability and fertilizer value of phosphate in SS-amended soil. Soil properties were the dominant factor determining plant available phosphate, where plant available phosphate decreased with increasing clay content, irrelevant of the type of treatment. There were, however, significant differences between the chemical and biologically removed SS, where the biologically removed SS had higher plant available phosphate. The RPFV % of the SS was comparable to MAP in terms of its plant availability. Wetting and drying cycles in the pot trial influenced the plant available phosphate from the SS, where the chemically treated sludge showed in general lower plant availability. The RPFV % of the biologically removed phosphate sludges was better than that of MAP and that of the chemically phosphate removed sludge were lower. Application of all the different sludge types resulted in a positive reaction on plant available phosphate for all the soils. All the trials were conducted at pH of about 5.5. However, it is expected that biologically P removed sludge will perform better in acid soils. The reason being that ferric phosphate in the chemically treated sludge is less soluble under pH conditions lower than 5.5 than above it. / Dissertation (MScAgric)--University of Pretoria, 2014. / tm2015 / Plant Production and Soil Science / MScAgric / Unrestricted

This master thesis gives an account for various aeration schemes that are utilized in conventional activated sludge treatment process in a wastewater treatment plant. By altering between different aeration schemes, the project aimed at reducing energy consumption along with maintaining the treatment performance at the plant. A series of experiments on the treatment performance over several different aeration schemes thus has been conducted on site at Solviken in Mora, Sweden. The project is basically a case study where no parameters were manually controlled except for the aeration schemes at the plant. Energy consumption reduction is logically relative to the ratio between aeration and non-aeration intervals. Several criteria were tested in regards to the treatment performance, including pH value, organic matters, phosphorus substances and microorganisms. The study has come up with a superior aeration scheme for the plant, as well as indicated processes that can be improved at the plant.

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In this paper, replacing complete or partly of growth mediums with secondarily wastewater was studied. Lipid content of Neochloris oleoabundans grown in a 0.3 X SE medium and autoclaved secondarily treated wastewater mixture was 22.27 % (w/w). The maximum biomass concentration of N. oleoabundans grown in wastewater with no additional nutrients was 0.636 g/L with 33% (w/w) glucose. Two culture lines, MA, and NA were isolated within our laboratory and could grow in secondarily treated wastewater with no additional nutrients. The maximum biomass concentration of MA in batch culture was 0.860 g/L and the sum of glucose and xylose was 40% (w/w). The maximum biomass concentration of NA was 1.562 g/l and the sum of glucose and xylose was 33.8% (w/w). The maximum specific growth rates of NA and MA were determined to be 0.0566 and 0.0337 per hour.

Microplastics are present in municipal wastewater treatment plant (WWTP) effluents; however, it is unclear whether these contaminants are ingested by biota living downstream of these outfalls. This study examined whether microplastic levels in caged biota, resident fish, and environmental samples were elevated near the Waterloo and Kitchener WWTP outfalls along the Grand River in the fall of 2019. Amphipods (Hyalella azteca), fluted-shell mussels (Lasmigona costata), and rainbow trout (Oncorhynchus mykiss) were caged at one upstream reference site and two impacted sites downstream of the Kitchener WWTP for 14 (amphipods and trout) or 28 (mussels) days. Rainbow darter (Etheostoma caeruleum) were collected using a backpack electrofisher from 10 sites up and downstream of both the Kitchener and Waterloo WWTPs, along with surface water and sediment samples. Whole body Hyalella, fish digestive tracts, and fluted-shell mussel tissues (hemolymph, digestive glands, and gills) were digested in 20% potassium hydroxide. Environmental samples were processed using filtration and density separation, then visual identification of microplastics was done. Elevated particle counts were found in rainbow trout digestive tracts at the Kitchener outfall site, compared to the upstream reference and downstream farfield sites. Additionally, particle concentrations in sediment were significantly higher at the Waterloo outfall, compared to all other sites (except for one upstream location). However, whole Hyalella, fluted-shell mussel tissues (hemolymph, digestive glands, and gills), digestive tracts of rainbow darter, and surface waters did not show elevated counts downstream of these discharges. Across all samples, fibers were the most common morphology, and blue and clear particles were prevalent in samples collected near WWTPs. Overall, these findings suggest that the Kitchener and Waterloo WWTPs could be important sources of particles to the Grand River, adding to our understanding of the fate of this contaminant in freshwater ecosystems. / Thesis / Master of Science (MSc)

This research addresses refractory forms of nitrogen that, even with advanced biological nitrification-denitrification systems are not removed completely from domestic wastewater. TKN (Total Kjeldahl Nitrogen), ammonia plus organic nitrogen, is one of the forms to measure the levels of nitrogen present in effluent wastewaters. Ferrate, a strong oxidant, was used for the treatment of these nitrogen forms with the objective of producing nitrogen compounds that can be removed by subsequent biological processes. Bench-scale experiments were performed on effluent samples taken prior to chlorination from an Orlando, FL wastewater treatment facility, using a biological nutrient removal process. The samples were treated with doses of ferrate ranging from 1 to 50 mg/L as FeO4–2 under unbuffered conditions. TKN removal as high as 70% and COD removal greater than 55% was observed. The TSS production after ferrate treatment was in a range of 12 to 200 mg/L for doses between 10 and 50 mg/L FeO4-2. After an optimum dose of ferrate was determined, three bench-scale reactors were operated under anoxic conditions for 10 to 12 days, two as duplicates containing the treated effluent and one as a control with untreated sample. Two different doses of ferrate were used as optimum dose for these experiments, 10 and 25 mg/L as FeO4-2. The purpose of these reactors was to determine the potential for biological removal of remaining nitrogen after ferrate oxidation of refractory nitrogen. Treated and raw samples were analyzed for Total Kjeldahl Nitrogen (TKN) (filtered and unfiltered), chemical oxygen demand (COD) (filtered and unfiltered), total suspended solids (TSS), nitrate (NO3-N), nitrite (NO2-N), and heterotrophic plate count (HPC). As a result, more than 70% of the soluble TKN was removed by chemical and biological oxidation for a sample treated with a dose of 25 mg/L FeO4-2, and less than 50% when treated with 10 mg/L FeO4-2. For the control samples run parallel to the ferrate treated samples, a maximum of 48% of soluble TKN and a minimum of 12% was removed. A three-log increase was observed in heterotrophic bacteria numbers for both doses during the operation of the reactors. Sodium ferrate was found to be an effective oxidant that can enhance the biodegradability of recalcitrant TKN present in municipal wastewaters. As mentioned before this research was develop using batch reactor units at bench-scale, therefore it is recommended to follow the investigation of the biodegradability of recalcitrant TKN of a ferrate treated sample under continuous flow conditions so that results can be extrapolated to a full-scale treatment facility.

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MSc Thesis - The effect of wastewater effluent on the gut microbiome of rainbow darter / The microbiome plays an important role in host physiology and can be influenced by species, diet, and environment. Municipal wastewater effluent contains a mixture of chemicals including antibiotics and antimicrobials that may affect the gut microbiome of fish living downstream of these discharges. Thus, this study examines the effect of wastewater treatment plant (WWTP) effluent on the gut microbiome of wild rainbow darter (Etheostoma cearuleum), and examines how the gut microbiome of wild fish changes in the lab. Fish were collected from sites upstream and downstream of 2 major WWTPs along the central Grand River and gut contents were aseptically sampled. After extracting gDNA, nested PCR of the V3-V4 region of the 16S rRNA gene, and Illumina sequencing were performed. The gut microbiome of exposed fish had increased bacterial diversity and was dominated by Proteobacteria, which has been linked to altered health outcomes in mammals. Next, rainbow darters were collected from a reference site on the Grand River. Fish were sampled in the field, after a 14 day lab acclimation, and after a 28 day exposure to environmental stressors (WWTP effluent or triclosan, an antimicrobial found in WWTP effluent). Surprisingly, there were no changes in the microbiome after exposure to environmental stressors. Major changes were observed between the field and laboratory fish suggesting that environment and diet are important factors influencing the gut microbiome. Changes in the gut microbiome continued up to 42 days in the lab, indicating longer acclimation periods may be needed. This study showed that effluents altered the gut microbiome of fish in the field, but not in the laboratory for unknown reasons. Laboratory studies indicated that transitioning to a new environment may require greater than 14 days before achieving a stable microbiome. / Thesis / Master of Science (MSc) / Wastewater is the largest source of pollution affecting Canada’s aquatic ecosystems; effluents contain antibiotics and antimicrobials that can affect fish and other aquatic life. The gut microbiome of fish is influenced by host species, its diet, and the environment, and thus contaminants released via wastewater effluents may alter the gut microbiome of fishes in receiving waters. This study found that the gut microbiota of rainbow darter fish exposed to wastewater effluents in the central Grand River (Waterloo/Kitchener, Ontario) were dominated by Proteobacteria and had increased diversity. Wild fish transitioned to the lab were dominated by Firmicutes and had decreased bacterial diversity in the gut compared to those in the wild. Altogether, these results suggest that wild fish exposed to wastewater effluents had altered gut microbiomes; transitions to new environments and laboratory acclimation periods are important considerations when studying the fish gut microbiome.