Vitellogenin in Pimephales promelas was used to assess estrogenicity of a local municipal effluent. Vitellogenin induction in male P. promelas increased in frequency and magnitude with increased exposure duration and was greater ("=0.05) than controls after 2 and 3 weeks of exposure. The level of vitellogenesis induced by effluent exposure was high compared to similar studies. A spring season evaluation followed. Biomarkers in P. promelas were used to assess the efficacy of a treatment wetland to remove toxicity and estrogenicity in final treated wastewater effluent. Comparisons were made with an effluent dominated stream and laboratory controls. Vitellogenin, GSIs (gonado-somatic indices), HSIs (hepato-somatic indices) and secondary sexual characteristics were biomarkers used in P. promelas models to assess aqueous estrogenicity. Biological indicators used to assess general fish health included hematocrit and condition factors. The estrogenic nature of the effluent was screened, concurrent with fish exposure, with GC/MS analysis for target estrogenic compounds including: 17-b estradiol, estrone, ethynylestradiol, Bisphenol A, nonylphenolic compounds, phthalates, and DDT. Plasma vitellogenin measured in P. promelas was significantly elevated (p < 0.0001) at the inflow site of the wetland and stream sites. GSIs for these exposures were less (a=0.001) at the wetland inflow site. At wetland sites closest to the inflow, secondary sexual charateristics, tubercle numbers and fat pad thickness, were less (a=0.0001). Hematocrit and condition factors were less (a=0.001) at sites closer to the wetland inflow. Seasonal variation was examined by repeating the effluent characterization in summer. Additionally, summer testing included exposure to an effluent dilution series. Fish condition heavily influenced interpretation of the results. Pre-acclimation exposure to spawning stresses may have altered many of the biological markers measured. Results are discussed relative to fish health and pre-exposure environment. Toxicity assessed with P. promelas biomarkers was compared with Ceriodaphnia dubia and Vibrio fischeri toxicty tests on this effluent. Biomarkers of fish health were somewhat less sensitive than C. dubia test endpoints, but more sensitive than V. fisheri.

The production of final effluents that meet discharged requirements and guidelines remain a major challenge particularly in the developing world with the resultant problem of surface water pollution. This study assessed the physicochemical and microbiological qualities of two wastewater treatment works in the Eastern Cape Province of South Africa in terms of the prevalence of faecal coliforms and Escherichia coli O157:H7 over a five month period. All physicochemical and microbiological analyses were carried out using standard methods. Data were collected in triplicates and analysed statistically using IBM SPSS version 20.0. The ranges of some of the physicochemical parameters that complied with set guidelines include pH (6.7 – 7.6), TDS (107 – 171 mg/L), EC (168 – 266 μS/cm), Temperature (15 – 24oC), NO3- (0 – 8.2 mg/L), NO2- (0.14 – 0.71 mg/L) and PO4 (1.05 – 4.50 mg/L). Others including Turbidity (2.64 – 58.00 NTU), Free Cl (0.13 – 0.65 mg/L), DO (2.20 – 8.48 mg/L), BOD (0.13 – 6.85 mg/L) and COD (40 – 482 mg/L) did not comply with set guidelines. The microbiological parameters ranged 0 – 2.7 × 104 CFU/100 ml for FC and 0 – 9.3 × 103 for EHEC CFU/100 ml, an indication of non-compliance with set guidelines. Preliminary identification of 40 randomly selected presumptive enterohemorrhagic E. coli isolates by Gram’s staining and oxidase test shows 100% (all 40 selected isolates) to be Gram positive while 90% (36 randomly selected isolates) were oxidase negative. Statistical correlation between the physicochemical and the microbiological parameters were generally weak except in the case of free chlorine and DO where they showed inverse correlation with the microbiological parameters. The recovery of EHEC showed the inefficiency of the treatment processes to effectively inactivate the bacteria, and possibly other pathogenic bacteria that may be present in the treated wastewater. The assessment suggested the need for proper monitoring and a review of the treatment procedures used at these treatment works.

Escherichia coli (E. coli) is a common inhabitant of surface waters in the developed and developing worlds. The majority of E. coli cells present in water are not particularly pathogenic to humans; however, there are some present in small proportion that possess virulence genes that allow them to colonize the digestive tract. Pathogenic E. coli causes acute and chronic diarrheal diseases, especially among children in developing countries and in travelers in these locales. The present study, conducted between August 2007 and July 2008, investigated the prevalence and distribution of virulent E. coli strains as either free or attached cells in the final effluents of three wastewater treatment plants located in the Eastern Cape Province of South Africa and its impact on the physico-chemical quality of the receiving water body. The wastewater treatment plants are located in urban (East Bank Reclamation Works, East London), peri-urban (Dimbaza Sewage Treatment Works) and in rural area (Alice Sewage Treatment Works). The effluent quality of the treatment plants were acceptable with respect to pH (6.9-7.8), temperature (13.8-22.0 °C), dissolved oxygen (DO) (4.9-7.8 mg/L), salinity (0.12-0.17 psu), total dissolved solids (TDS) (119-162 mg/ L) and nitrite concentration (0.1-0.4 mg/l). The other xii physicochemical parameters that did not comply with regulated standards include the following: phosphate (0.1-4.0 mg/L); chemical oxygen demand (COD) (5-211 mg/L); electrical conductivity (EC) (237-325 μS/cm) and Turbidity (7.7-62.7 NTU). Results suggest that eutrophication is intensified in the vicinity of the effluent discharge points, where phosphate and nitrate were found in high concentrations. Presumptive E. coli was isolated from the effluent samples by culture-based methods and confirmed using Polymerase Chain Reaction (PCR) techniques. Antibiogram assay was also carried out using standard in vitro methods on Mueller Hinton agar. The viable counts of presumptive E. coli for the effluent samples associated with 180 μm plankton size ranged between 0 – 4.30 × 101 cfu/ml in Dimbaza, 0 – 3.88 × 101 cfu/ml in Alice and 0 – 8.00 × 101 cfu/ml in East London. In the 60 μm plankton size category E. coli densities ranged between 0 and 4.2 × 101 cfu/ml in Dimbaza, 0 and 2.13 × 101 cfu/ml in Alice and 0 and 8.75 × 101 cfu/ml in East London. Whereas in the 20 μm plankton size category presumptive E. coli density varied from 0 to 5.0 × 101 cfu/ml in Dimbaza, 0 to 3.75 × 101 cfu/ml in Alice and 0 to 9.0 × 101 cfu/ml in East London. The free-living presumptive E. coli density ranged between 0 and 3.13 × 101 cfu/ml in Dimbaza, between 0 and 8.0 × 101 cfu/ml in Alice and between 0 and 9.5 × 101 cfu/ml in East London. Molecular analysis successfully amplified target genes (fliCH7, rfbEO157, ial and aap) which are characteristic of pathogenic E. coli strains. The PCR assays using uidA-specific primer confirmed that a genetic region homologous in size to the E. coli uidA structural gene, including the regulatory region, was present in 3 of the E. coli isolates from Alice, 10 from Dimbaza and 8 from East London. Of the 3 E. coli isolates from Alice, 1 (33.3%) was positive for the fliCH7 genes and 3 was positive for rfbEO157 genes. Out of the 10 isolates from Dimbaza, 4 were xiii positive for fliCH7 genes, 6 were positive for the rfbEO157 genes and 1 was positive for the aap genes; and of the 8 isolates from East London, 1 was positive for fliCH7 genes, 2 were for the rfbEO157 genes, 6 were positive for the ial genes. Antimicrobial susceptibility profile revealed that all of the E. coli strains isolated from the effluent water samples were resistant (R) to linezolid, polymyxin B, penicillin G and sulfamethoxazole. The E. coli isolates from Dimbaza (9/10) and East London (8/8) respectively were resistant to erythromycin. All the isolates were found to be susceptible (S) to amikacin, ceftazidime, ciprofloxacin, colistin sulphate, ceftriaxone, cefotaxime, cefuroxime, ertapenem, gatifloxacin, gentamycin, imidazole, kanamycin, meropenem, moxifloxacin, neomycin, netilmicin, norfloxacin and tobramycin. The findings of this study revealed that the Alice wastewater treatment plant was the most efficient as it produced the final effluent with the least pathogenic E. coli followed by the Dimbaza wastewater treatment plant. In addition, the findings showed that the wastewater treatment plant effluents are a veritable source of pathogenic E. coli in the Eastern Cape Province watershed. We suggest that to maximize public health protection, treated wastewater effluent quality should be diligently monitored pursuant to ensuring high quality of final effluents.

Chorine is used by the Village Creek Waste Water Treatment Plant to kill pathogenic microorganisms prior to discharge of the effluent into the Trinity River. The residual chlorine in the river impacted aquatic life prompting the U.S. Environmental Protection Agency in December 1990 to require dechlorination using sulfur dioxide. One pre-dechlorination and four post-dechlorination assessments of phytoplankton, periphyton, and zooplankton communities were conducted by the Institute of Applied Sciences at the University of North Texas. Dechlorination had no effect on the phytoplankton community. The periphyton community exhibited a shift in species abundance with a more even distribution of organisms among taxa. No change occurred in zooplankton species abundance, however, there was a decrease in zooplankton density following dechlorination.

A toxicological study was initiated because of concerns about allegations that the Texas Water Commission that effluent from the Dallas Central Wastewater Treatment Plant, which discharges into the Trinity River, was affecting downstream water quality. Monthly, flow-weighted composite effluent samples were collected. Grab samples were also collected upstream and downstream from the effluent from April 1989 to August 1991. Toxicity tests were conducted on these samples using Ceriodaphnia dubia as the test organism. Samples were collected four times during this study in which rainfall occurred prior to sampling. In every instance, this "first flush" of the watershed during a rising hydrograph was toxic to C. dubia upstream. Analyzing toxicity by season resulted in a statistically significantly lower neonate production in the effluent than in the river samples during the months of June, July, and August. This impact on neonate production was suspected of being caused by organic pesticides which are used for insect control on lawns. The effluent was never acutely toxic to C. dubia. Primarily, toxic occurrences in either the effluent or the river samples were primarily of a chronic nature. Overall, survival of C. dubia was affected more frequently at the upstream site than in the effluent or the downstream site. Because EPA's Phase I Acute Toxicity Identification Evaluations (TIEs) methods were designed for identifying acute toxicity, two alternative strategies were attempted to identify chronic toxicity. The first attempt was the modification of the phase I acute TIE methodologies. This was done by processing more sample through the phase I characterization tests. This approach was inadequate due to toxicity that occurred during the last several days of the seven-day C. dubia reproduction test. The second strategy for identifying chronic toxicity within a TIE involved the use of freeze concentration. During this preliminary investigation ofthe efficiency of freeze concentration, four metals and two organic compounds were freeze concentrated.

The impact of a power plant discharge (Clinch River Plant, CRP, Carbo, Virginia) on resident snail populations was assessed. In 1988, snail absence below the plant, was attributed to plant discharges rather than naturally occurring habitat limitations. Habitat limitations for the two dominant snail species, Leptoxis praerosa and Pleurocera unciale were defined before power plant impact was assessed. Eleven physicochemical parameters (i.e., flow rate, substrate type, silt accumulation, depth, water chemistry and food biomass parameters) were measured at selected sites and compared to snail density. Flow rate, substrate type and periphyton biomass were the most influential parameters in determining Leptoxis density; while periphyton biomass was the most influential for Pleurocera. Cluster analysis also linked Leptoxis density with river structure and flow. Other variables linked to Pleurocera density were flow rate, river structure and silt. Although Leptoxis is most prolific in riffle/shoal areas and Pleurocera in slower riffle-pool interfaces, these two ‘species often coexist. This research suggests that habitat partitioning between these two species is influenced most by flow rate. Greatest density of Leptoxis occurred at flow rates of 20-30 cm/sec. Frequency of occurrence was greatest at 20-100 cm/sec. Pleurocera occurred most frequently at flow rates of 20-30 cm/sec with greatest density at 25-45 cm/sec. Measurements of impact of the CRP effluent (i.e., toxicity, metals {mainly copper} bioaccumulation in aufwuchs and snails, and cellulase enzyme activity impairment) were summarized by using zone delineation. Habitat parameters were measured below plant discharges and upstream, and compared with water column Cu, snail tissue Cu and aufwuchs Cu measurements. Habitat selection was strongly influenced by effluent but the role of waterborne metals concentration and habitat alterations (e.g. periphyton changes and bioconcentration) was unclear. Feeding studies were conducted to estimate impact of aufwuchs bioconcentration of metals on snails. Leptoxis significantly bioconcentrated Cu when fed aufwuchs containing 564 (±269) ug Cu/g in artificial stream feeding studies, but no cellulase impairments were seen in these studies. No foodborne bioconcentration was found from aufwuchs containing up to 20,000 (±18,400) ug Zn/L. These results suggest that though foodborne uptake of Cu may occur, water column Cu concentrations may have to be an order of magnitude higher for impairment to occur through injestion than through waterborne exposures. Acute and chronic effects of both whole effluent and Cu on Leptoxis were measured in laboratory and artificial stream exposures. The 96-hr LC₅₀ was 95% effluent (containing 148 ug Cu/L)in flow-through exposures, but in Static stirred exposures, 100% effluent (105 ug Cu/L)was not toxic. The lowest-observable effect concentration (LOEC)from 30-day exposures was 10% effluent (22 ug Cu/L) causing cellulase activity impairment (70% of control activity) and bioconcentration (300 ug Cu/g). Constituents of effluent other than Cu were believed to contribute to impairment effects since no impairment was found in 30-day CuSO₄ dosings of up to 25 ug Cu/L. The LOEC for Cu from 30-day CuSO₄ dosings ranged from 17-35 ug/L and the no-observed effect concentration (NOEC)was 12 ug Cu/L. The EPA water quality criteria concentration (17 ug Cu/L)was questionable for Leptoxis in long-term exposures (114-day), causing enzyme impairment and mortality. Chronically toxic conditions to Leptoxis occurred on the left side of the river for 0.7 km downstream of discharge, where the water column contained 42 ug Cu/L, while acutely toxic conditions occurred in the immediate mixing zone. Artificial stream impairment tests were substantiated in the river except in lower reaches of the impairment zone (left side of river, 0.7-0.9 km below cooling tower discharge), where snail absence was attributed to periphyton Cu bioconcentration (242 ug Cu/g). Functional recovery (of enzyme activity) was found at the next acceptable habitat downstream (Station 14A), so the area of impact extended 0.9 km downstream of the discharge on the left side of the river. It was concluded that zone delineation by simultaneously evaluating structural and functional aspects of environmental change is a better approach to impact assessment than approaches that only use functional measurements. / Ph. D.

This study evaluated the effects of a major chlorine disturbance on fish communities in Pecan creek by the City of Denton's Pecan Creek Water Reclamation Plant. Fish communities in Pecan Creek were sampled using a depletion methodology during February, April, July, and November, 1999. February and April sampling events showed that the fish communities were severely impacted by the chlorine. Sampling during July and November showed fish communities recovered in Pecan Creek. The first-twenty minutes of shocking and seining data were analyzed to mirror an equal effort methodology. This methodology was compared to the depletion methodology to see if the equal effort methodology could adequately monitor the recovery of Pecan Creek after the chlorine disturbance. It was determined that the equal effort methodology was capable of monitoring the recovery of Pecan Creek, but could not accurately represent the fisheries community as well as the depletion method. These data using the twenty-minute study were compared to a previous study. Results of this study were similar to those found in a previous study, although fish communities were more severely impacted and took longer to recover.

NCRP report No.160 states that medical exposure increased to nearly half of the total radiation exposure of the U.S. population from all sources in 2006 (NCRP 2009). Part of this increase in exposure is due to the rise in nuclear medicine procedures. With this observed growth in medical radionuclide usage, there is an increase in the radionuclide being released into wastewater after the medical procedures. The question then arises: what is the behavior of medical radionuclides and their impurities in the wastewater process? It is important to note that, often, medical radionuclides are not exactly 100% radionuclide pure, but they meet a certain standard of purity. Of particular interest are the longer lived impurities associated with these medical radionuclides. The longer lived impurities have a higher chance of reaching the environment. The goal of this study is to identify the behavior of medical radionuclides and their impurities associated with some of the more common radiopharmaceuticals, including Tc-99m and I-131, and locate and quantify levels of these impurities in municipal wastewater and develop a model that can be used to estimate potential dose and risk to the public. / Graduation date: 2012 / Access restricted to the OSU Community at author's request from May 24, 2012 - May 24, 2014

Separation of dissolved metals from aqueous solutions using liquid membrane technology is highly advantageous owing to the degree of separation achieved, efficiency and application potential. In the present investigation four types of liquid membranes - bulk liquid membrane (BLM), emulsion liquid membrane (ELM), electrostatic pseudo liquid membrane (ESPLIM) and unified liquid membrane (ULM) have been extensively studied, for their application in extraction and concentration of dissolved metals from effluents. Experiments were conducted with various metal systems to optimize both system and process conditions and to find out the effect of various parameters on the performance of the process. Different mass transport models were proposed for each type, taking diffusional and kinetic resistances into account. Models were extended for simultaneous extraction systems and were verified by different metal-carrier experiments. Good agreement was found between the concentration profiles obtained from the models and the experimental data, thereby establishing the validity of models for all the four types of liquid membranes. The stirred cell employed in BLM process eliminates emulsification and demulsification processes. It also provides simultaneous contact of the organic liquid membrane phase with aqueous feed and strip phases. Overall rate expressions for extraction and stripping in BLM are based on an assumed kinetic mechanism to explain the process qualitatively. It was found that the magnitude^ of diffusional and kinetic resistances determines the overall mass transfer coefficient. The relative magnitude of mass transfer coefficient, reaction rate constants and equilibrium constants enables to visualize the controlling regime of the process. The problem of low flux rate due to high diffusion resistances, inefficient operation and exorbitant costs encountered in bulk and supported liquid membranes (SLM) are overcome in an ELM. In the ELM process, an emulsion of organic membrane phase and aqueous inner phase, is dispersed in the continuous aqueous feed phase. This gives a highly selective and ultra thin liquid film generating a large mass transfer area for separation. Experimental results on membrane instability and emulsion swelling indicate that volumetric leakage rate depends linearly on the stirring speed and that the nature of surfactant does not have any appreciable effect on emulsion swelling. A general permeation model was developed taking into account the external mass transfer around the emulsion drop, diffusion in the drop, reaction at the aqueous-organic interface, leakage of the internal phase to the external phase due to membrane breakup and emulsion swelling due to osmotic pressure difference. Model equations with appropriate boundary conditions were numerically solved by orthogonal collocation technique for a set of model parameters obtained either from known correlations or from independent experiments. Comparison of the model predictions with experimental data from the batch permeation of chromium and other metals using carrier Alamine 336 or LIX 64N, shows that the model predictions are in very good agreement with the experimental findings. Further this model can be used to simulate the effects of various experimental conditions such as metal and hydrogen ion concentrations, carrier concentration, drop diameters, etc., for similar systems. Studies on ESPLIM were conducted with the aim of demonstrating the effectiveness of this new separation process and to develop a simple transport model for metal permeation. In the ESPLIM process, a high electrical field (3-5 kV A.C.) is used for phase dispersion. This system consists of a rectangular reactor filled with membrane solution divided into extraction and stripping cells by a centrally placed integrated type baffle which also acts as an electrode. Two more electrodes were placed in the extraction and stripping cells, where feed and strip phases are introduced from the top of the reactor. When high electrical field is applied across the electrodes, fine droplets of feed and strip are formed and are dispersed in extraction and stripping cells where simultaneous extraction and stripping occurs. The process can be viewed as simultaneous counter current extraction and stripping. The aqueous drops coalesce in the settlers at the bottom of the reactor and are removed continuously. Steady state mass transport model proposed for ESPLIM system accounts for the vertical counter-current extraction and stripping processes taking place in the extraction and stripping cells, together with the lateral transport process of the metal-complex and carrier across the two cells through the integrated baffle zone. The model equations were solved analytically to obtain concentration profiles as a function of the height of the reactor. The required parameters such as mass transfer coefficients, diffusion coefficients etc. were estimated using different correlations. Model predictions agreed remarkably well with the experimental data under various process conditions. From this investigation, it was found that ESPLIM is a simple, efficient and economical process and can be applied in a variety of situations. Based on a suitable combination of solvent extraction, dispersion and liquid membrane technique, a new type of separation system called " Unified Liquid Membrane " was developed. The ULM unit was designed and fabricated, and experiments were conducted to evaluate its performance. The ULM is basically derived from ESPLIM by changing the reactor, baffle design and dispersion technique. Aqueous feed and strip phases were atomized using compressed air through a fine nozzle and are dispersed on either side of an integrated baffle plate that divides the reactor into extraction and stripping cells. Tapering bottom of the reactor reduces the dead volume of the liquid in the settlers and the baffle plate remarkably reduces the leakage problem as well as the resistance through the baffle. Experiments were conducted using LIX 64N and Alamine 336 as carriers for copper and chromium and / or zinc. Mass transport model proposed considers both chemical and phase equilibria in extraction and stripping cells, vertical and lateral transport of carrier and complex across the extraction and stripping cells through the baffle zone. The model equations were solved using initial conditions at the top of the reactor, and equilibrium data for extraction and stripping cells. Effect of various experimental conditions and process parameters was simulated using this model and the model predictions are found to be in excellent agreement with the experimental data. The ULM system developed in this investigation overcomes the major limitations encountered with the other types of liquid membranes while retaining all the advantages of this technology. The problem of high mass transfer resistance from bulk phase to metal permeation as in the case of BLM was eliminated by good phase dispersion. Additional resistance to mass transport from solid membrane as in the case of SLM was removed by using an integrated baffle which also avoids problems of membrane instability, pore clogging and selectivity. The complex problems of emulsification and demulsification were completely eliminated making the system much simpler and efficient. Very good phase dispersion was obtained by atomization without the need for either stirring the whole system or application of high electrical field in the reactor. The membrane liquid within the integrated baffle elements allows easy transport of different species between extraction and stripping cells while completely preventing the mixing of the two aqueous phases. The problems of leakage, swelling and occlusion were avoided due to very short residence time of the aqueous drops in the reactor. It was found that the new ULM configuration is simple, elegant, highly efficient and superior to the other types of liquid membrane systems.

Water is an indispensible and yet a difficult resource to be renewed, thus water scarcity has become one of the major challenges faced worldwide, with the Southern regions of Africa being the most impacted and affected, especially the Eastern Cape Province of South Africa where rural communities depend on receiving waterbodies that are often negatively impacted by wastewater final effluents. This present study was conducted between August and December 2010 to assess the physicochemical and microbial qualities of the final effluents of peri-urban and rural communities based wastewater treatment plants in the Eastern Cape Province. The physicochemical parameters were determined on site and in the laboratory, while bacteriological qualities were determined using culture based techniques. The virological qualities were determined by molecular methods using reverse transcriptase polymerase chain reaction for the target RNA virus and the conventional polymerase chain reaction for the target DNA virus. For both wastewater treatment plants, the physicochemical parameters ranged as follows: chemical oxygen demand (5.95-45 mg/L); total dissolved solids (114.5-187.0 mg/L); salinity (0.12-0.20 psu); temperature (14.2-25.7oC); pH (6.0-7.6); nitrate and nitrites (1.55-6.7 mg/L and 0.023-1.15 mg/L respectively); biological oxygen demand (3.5-7.8 mg/L); turbidity (1.49-6.98 NTU); and chlorine residual (0-2.97 mg/L). Feacal indicator bacteria counts ranged as follows: feacal coliforms (0-1.25×104 cfu/100 ml); total coliforms (0-3.95×104 cfu/100 ml); and enterococci (0-5.0×103 cfu/100 ml). xviii Seventy five percent of the rural community based plant and 80 percent of the peri-urban community based plant were positive for coxsackie A virus, while hepatitis A virus was detected in all the rural community based plant 80 percent of the peri-urban community based plant. This study suggests the need for intervention by appropriate regulatory agencies to ensure regular monitoring of the qualities of final effluents of wastewater treatment facilities in the Eastern Cape Province and ensure compliance to established guidelines.