Treatment of waste sludge has become an important issue in recent years around the world. However, the trend of waste sludge treatment has shifted from volume minimization and stabilization to reuse of the sludge and recover the energy potential of it. Therefore, anaerobic treatment of sludge is gaining popularity because of byproduct methane production and high percentage of VSS reduction. Pre-treatment of sludge before anaerobic digestion in order to increase methane production, and ozone pre-treatment in this context, is one such option. Domestic sludge also contains the recently recognized, so called, emerging compounds such as Endocrine Disrupting Compounds (EDCs). Therefore treatment of EDCs in sludge is another challenge in waste sludge treatment since direct discharge of such chemicals may harm the environment by causing gender shifts within the fauna. In this context two hormones (estrone and progesterone), three pharmaceuticals (acetaminophen, carbamazepine and diltiazem) and one plasticizer (benzyl-butyl phthalate) were routinely analyzed in sludge samples which were subjected to treatment during this study. Treatment of EDCs during anaerobic digestion and the effect of ozonation both on the performance of digestion and the treatability of EDCs were investigated in this study. Four 2.5L anaerobic jars were used for anaerobic digestion connected to four 1L plastic graduated cylinders immersed in salt-water to collect the off gas. Anaerobic sludge culture of the reactor and the sludge feed to the reactors were obtained from Ankara Tatlar Wastewater Treatment Plant anaerobic digester and return activated sludge (RAS) line, respectively. One of the anaerobic digesters was used as control (no ozonation) and the others were fed with sludge samples ozonated at three different ozone doses 0.65, 1.33 and 2.65 mg ozone/g biomass. Sludge ages of the reactors were initially set to 25 days and the reactors were fed once every 2 days. The TSS, VSS, total gas volume, COD, pH, CH4 percentage and EDCs were analyzed routinely. In the reactors, operated at 25 days, because of the observation of reduction of TSS, SRT was set to infinity / thus, sludge wastage was terminated. Following the startup it was seen that at 2.65 mg ozone/g biomass dose TSS and VSS did not stay constant in the reactor and dropped sharply in the course of operation, indicating that system was not steady at this SRT. However, upon stoppage of sludge wastage from the reactors, thereby setting SRT to infinity, a steady culture could be maintained in the reactors. Both total gas production and CH4 percentage increased with the increasing doses of ozone with respect to control reactor. For 2.65 mg/g ozonated reactor total gas volume doubled the amount produced in the control reactor. All the EDCs within the scope of this study were analyzed in sludge using ultrasound-aided sequential sludge extraction method twice a week and the results showed that ozonation affected treatment of EDCs for up to 96%. The highest removal rate was obtained with natural hormones. Rates of treatment of pharmaceuticals were the second best.

Treatment of waste sludge has become an important issue in recent years around the world. However, the trend of waste sludge treatment has shifted from volume minimization and stabilization to reuse of the sludge and recover the energy potential of it. Therefore, anaerobic treatment of sludge is gaining popularity because of byproduct methane production and high percentage of VSS reduction. Pre-treatment of sludge before anaerobic digestion in order to increase methane production, and ozone pre-treatment in this context, is one such option. Domestic sludge also contains the recently recognized, so called, emerging compounds such as Endocrine Disrupting Compounds (EDCs). Therefore treatment of EDCs in sludge is another challenge in waste sludge treatment since direct discharge of such chemicals may harm the environment by causing gender shifts within the fauna. In this context two hormones (estrone and progesterone), three pharmaceuticals (acetaminophen, carbamazepine and diltiazem) and one plasticizer (benzyl-butyl phthalate) were routinely analyzed in sludge samples which were subjected to treatment during this study. Treatment of EDCs during anaerobic digestion and the effect of ozonation both on the performance of digestion and the treatability of EDCs were investigated in this study. Four 2.5L anaerobic jars were used for anaerobic digestion connected to four 1L plastic graduated cylinders immersed in salt-water to collect the off gas. Anaerobic sludge culture of the reactor and the sludge feed to the reactors were obtained from Ankara Tatlar Wastewater Treatment Plant anaerobic digester and return activated sludge (RAS) line, respectively. One of the anaerobic digesters was used as control (no ozonation) and the others were fed with sludge samples ozonated at three different ozone doses 0.65, 1.33 and 2.65 mg ozone/g biomass. Sludge ages of the reactors were initially set to 25 days and the reactors were fed once every 2 days. The TSS, VSS, total gas volume, COD, pH, CH4 percentage and EDCs were analyzed routinely. In the reactors, operated at 25 days, because of the observation of reduction of TSS, SRT was set to infinity / thus, sludge wastage was terminated. Following the startup it was seen that at 2.65 mg ozone/g biomass dose TSS and VSS did not stay constant in the reactor and dropped sharply in the course of operation, indicating that system was not steady at this SRT. However, upon stoppage of sludge wastage from the reactors, thereby setting SRT to infinity, a steady culture could be maintained in the reactors. Both total gas production and CH4 percentage increased with the increasing doses of ozone with respect to control reactor. For 2.65 mg/g ozonated reactor total gas volume doubled the amount produced in the control reactor. All the EDCs within the scope of this study were analyzed in sludge using ultrasound-aided sequential sludge extraction method twice a week and the results showed that ozonation affected treatment of EDCs for up to 96%. The highest removal rate was obtained with natural hormones. Rates of treatment of pharmaceuticals were the second best.

Treatment of waste sludge has become an important issue in recent years around the world. However, the trend of waste sludge treatment has shifted from volume minimization and stabilization to reuse of the sludge and recover the energy potential of it. Therefore, anaerobic treatment of sludge is gaining popularity because of byproduct methane production and high percentage of VSS reduction. Pre-treatment of sludge before anaerobic digestion in order to increase methane production, and ozone pre-treatment in this context, is one such option. Domestic sludge also contains the recently recognized, so called, emerging compounds such as Endocrine Disrupting Compounds (EDCs). Therefore treatment of EDCs in sludge is another challenge in waste sludge treatment since direct discharge of such chemicals may harm the environment by causing gender shifts within the fauna. In this context two hormones (estrone and progesterone), three pharmaceuticals (acetaminophen, carbamazepine and diltiazem) and one plasticizer (benzyl-butyl phthalate) were routinely analyzed in sludge samples which were subjected to treatment during this study. Treatment of EDCs during anaerobic digestion and the effect of ozonation both on the performance of digestion and the treatability of EDCs were investigated in this study. Four 2.5L anaerobic jars were used for anaerobic digestion connected to four 1L plastic graduated cylinders immersed in salt-water to collect the off gas. Anaerobic sludge culture of the reactor and the sludge feed to the reactors were obtained from Ankara Tatlar Wastewater Treatment Plant anaerobic digester and return activated sludge (RAS) line, respectively. One of the anaerobic digesters was used as control (no ozonation) and the others were fed with sludge samples ozonated at three different ozone doses 0.65, 1.33 and 2.65 mg ozone/g biomass. Sludge ages of the reactors were initially set to 25 days and the reactors were fed once every 2 days. The TSS, VSS, total gas volume, COD, pH, CH4 percentage and EDCs were analyzed routinely. In the reactors, operated at 25 days, because of the observation of reduction of TSS, SRT was set to infinity / thus, sludge wastage was terminated. Following the startup it was seen that at 2.65 mg ozone/g biomass dose TSS and VSS did not stay constant in the reactor and dropped sharply in the course of operation, indicating that system was not steady at this SRT. However, upon stoppage of sludge wastage from the reactors, thereby setting SRT to infinity, a steady culture could be maintained in the reactors. Both total gas production and CH4 percentage increased with the increasing doses of ozone with respect to control reactor. For 2.65 mg/g ozonated reactor total gas volume doubled the amount produced in the control reactor. All the EDCs within the scope of this study were analyzed in sludge using ultrasound-aided sequential sludge extraction method twice a week and the results showed that ozonation affected treatment of EDCs for up to 96%. The highest removal rate was obtained with natural hormones. Rates of treatment of pharmaceuticals were the second best.

Evidence from an abundant number of studies suggests that human female reproductive functions have become impaired over the past half century and that there might be a relationship between endocrine disrupting chemicals (EDCs) and reduced fertility. It is, however, not known by what mechanisms EDCs affect different reproductive functions such as endometrial receptivity, embryo implantation and placentation. The endometrium is continuously changing its morphological and functional properties, responding to cyclic changes of oestrogen and progesterone levels during the menstrual cycle. These changes include monthly preparation for embryo implantation through changed endometrial angiogenic activity and consequent changes in endometrial vasculature. Use of primary human endometrial endothelial cells (HEECs) in this work was evaluated as a possible screening tool for effects caused by EDCs on human endometrial vasculature and subsequently on various endometrial functions. In this study HEEC and endometrial stromal cells were isolated. HEECs were grown in monocultures, and together with stromal cells in co-cultures, and exposed to endocrine active substances. These were cadmium, which has oestrogenic properties, tamoxifen, with anti-oestrogenic effects, mifepristone, which is an anti-progestin, and bisphenol A, with oestrogenic properties. The effects were evaluated by using proliferation and viability assays, migration and tube formation assays, quantitative PCR (qPCR), immunohistochemistry and western blot. Cadmium affected the expression of angiogenesis-related genes, and caused different effects in HEECs cultured alone vs. HEECs co-cultured with stromal cells. Tamoxifen altered the expression of angiogenesis-related genes and reduced HEEC migration, thus having an anti-angiogenic effect. Mifepristone caused reduced formation of tubular structures in tube-formation assays involving HEECs co-cultured with stromal cells. Bisphenol A promoted tube formation in co-cultured HEECs which was related to changes in the expression of several angiogenesis-related genes as well as up-regulated expression of VEGF-D protein. In conclusion, we showed that EDCs have the ability to induce changes in endometrial angiogenic activity in vitro and may thus disturb normal endometrial functions related to fertility and pregnancy. HEECs grown in vitro may provide valuable information on the effects of EDCs on human endometrial functions. However, this model is not suitable as a large-scale screening tool.

Alkylphenol polyethoxylates (APEOs), a class of nonionic surfactants, have been widely used for industrial, agricultural and household applications. The biodegradation metabolites of APEOs, such as nonylphenol and octylphenol, are more persistent and known to disrupt endocrine function in wildlife and human. These compounds are also recognized as endocrine disrupting chemicals (EDCs). The objectives of this study were to investigate the distribution and removal efficiencies of EDCs, including nonylphenol diethoxylates (NP2EO), nonylphenol monoethoxylates (NP1EO), nonylphenol (NP) and octylphenol (OP), of wastewater treated by the constructed wetland systems along the Dahan River and around the Dapeng Bay, respectively. In addition, the method of risk quotient was used to evaluate the potential ecological risk of APEOs to aquatic organisms in current study. The water samples collected from 32 sampling sites in the Dapeng Bay National Scenic Area including Datan, Pengcun and Linbain right bank constructed wetlands. The samples were then concentrated by solid phase extraction, and analyzed for target compounds by HPLC/fluorescence. According to the results, nonylphenol diethoxylates, nonylphenol monoethoxylates, nonylphenol and octylphenol were found approximately equal to 29.9, 47.3, 20.5 and 57.7 %, respectively of the samples from three constructed wetlands with concentrations ranged from <3.3 to 968.7, <3.3 to 226.5, <1.3 to 238.4 and <1.0 to 1458.7 ng/L, respectively. Temporal variation of APEOs showed a decreasing in the order of summer¡Öspring¡Öwinter¡Öautumn. The removal efficiencies of APEOs in these constructed wetlands showed a decreasing order of Datan wetland¡ÖPengcun wetland¡ÖLinbain right bank wetland. In addition, the samples collected from 18 sampling sites from the constructed wetlands along the riparian of Dahan River including Daniaopi, Hsin-Hai Bridge Phases I and II constructed wetlands. According to the results, nonylphenol diethoxylates, nonylphenol monoethoxylates, nonylphenol and octylphenol were found approximately equal to 91.9¡B84.8¡B17.1 and 73.7 %, respectively of samples collected from three constructed wetlands with concentrations ranged from <3.3 to 11191.5, <3.3 to 6069.0, <1.3 to 671.0 and <1.0 to 5581.9 ng/L, respectively. The removal potential of APEOs in these constructed wetlands showed a decreasing order of Hsin-Hai Bridge Phases II¡ÖDaniaopi¡ÖHsin-Hai Bridge Phases I constructed wetland. Regarding the ecological assessment in this research, the calculated risk quotients were up to 30 times higher in the constructed wetland systems of Dahan River than those in the Dapeng Bay National Scenic Area, indicating that the existing concentrations of these EDCs in wetland systems might cause potential ecological risks to aquatic organisms. Furthermore, the decreasing risk quotient from influent to effluent indicating the capabilities of treating alkylphenolic compounds in these constructed wetlands.

abstract / Biological Sciences / Doctoral / Doctor of Philosophy

Phytosterols are naturally occurring compounds which regulate membrane fluidity and serve as hormone precursors in plants. They also have the potential to cause endocrine disturbances in aquatic animals at concentrations as low as 10 µg/L. Wastewaters from several industries which process plant matter can contain phytosterols at concentrations in excess of the above-stated level. Despite their endocrine disruption potential, very little is known about phytosterol physical properties and their biotransformation potential in biological treatment systems. Aerated stabilization basins (ASBs) are common biological treatment systems in North American pulp and paper mills. ASBs are large open lagoons which use tapered surface aeration to remove COD and prevent sulfate reduction in the water column. Phytosterols are released from wood during the pulping process and a small fraction enters the wastewater stream during washing of the pulp. Therefore, phytosterols may be exposed to aerobic or anaerobic environments depending on their solubility and solid-liquid partitioning behavior. The overall objective of this research was to systematically and quantitatively assess the biotransformation potential of phytosterols in biological treatment systems and to examine conditions leading to reduction of these compounds in wastewater effluent streams. The results of this research showed that phytosterols are sparingly soluble with aqueous solubility below 1 µg/L when present as a mixture. Phytosterols have a strong affinity to adsorb to solids and dissolved organic matter. The affinity for aerobic biomass was greater than for wastewater solids. The stigmasterol desorption rate and extent from wastewater solids increased with an increase in pH from 5 or 7 to 9. Phytosterols were biotransformed under aerobic conditions but not under sulfate-reducing or methanogenic conditions by stock cultures developed in this study. Biotransformation under nitrate-reducing conditions could not be confirmed conclusively. The continuous-flow system was successful in removing 72 to 96% of phytosterols. Biotransformation accounted for 23, 14 and 41 % of campesterol, stigmasterol and β-sitosterol removal, respectively. Phytosterols accumulated in the reactor sediment and accounted for 97 % of the total phytosterols remaining in the system. Phytosterols can be removed from wastewater streams during biological treatment by a combination of biotransformation and solids partitioning and control of system pH, DO and available carbon and energy sources can increase the degree of phytosterols removal. The results of this research can be used to engineer effective biological treatment systems for the removal of phytosterols from pulp mill wastewaters and other phytosterol-bearing wastewater streams.

Over the past several decades, there has been growing concern among the public and scientific community regarding adverse health effects resulting from exposure to natural and synthetic compounds that act as endocrine disrupters. The structural similarity of many of these compounds to natural hormones and receptors, as well as their ubiquity in the environment, can result in the potential for interference with the endocrine system of wildlife and humans. Much of the research examining the adverse effects of wildlife exposure to endocrine disrupting chemicals (EDCs) has focused on effects on reproduction or short-term changes in hormone physiology. However, organisms exposed to low levels of EDCs at early life stages could also be susceptible to developmental effects, including neurological and other physiological changes affecting later life stages. In birds, migration can be an important component of the annual life cycle and it can be vulnerable to disruption given that it is under endogenous hormonal and neurological control. Previous studies have shown that developmental exposure of birds to thyroid hormone disruptors, such as polychlorinated biphenyls (PCBs), have resulted in reduced hatching success, lower growth rates, teratogenicity, impaired development, and immunotoxicity. In this thesis, I aimed to supplement what is currently known regarding the effects of developmental exposure to low levels of a mixture of endocrine disrupting chemicals in songbirds, as well as further investigate the latent consequences of such an exposure on migratory life stages. I initially investigated the potential physiological and developmental effects of early exposure to Aroclor 1254, a PCB mixture, in two passerine songbird species: European starlings (Sturnus vulgaris) and red-winged blackbirds (Agelaius phoeniceus) during the critical nestling period. In 2011, European starlings and red-winged blackbirds were orally administered Aroclor 1254 over the period of nestling development, which was repeated in 2012 with only European starlings. For both years, morphological parameters (body mass, tarsus, wing-chord and bill-lengths) were measured daily and plasma thyroid hormones were measured. Additional measurements of wing chord and tarsus length fluctuating asymmetry (FA) were taken in the second year, to further assess contaminant-induced alterations in developmental stability. I found that treatment with environmentally-relevant levels of Aroclor 1254 caused increasing liver residues above the controls but did not result in overt effects on morphological growth parameters during the nestling period in either starlings or red-winged blackbirds. However, we did observe significant differences in 2012 starling’s wing chord FA at day 10 and 13, and tarsus length FA between all treatment groups and controls, indicating the potential for PCB-induced stress. Nestling thyroid hormone profiles (T3) sampled throughout the nestling period supported developmental changes but did not reveal any differences among treatment groups. Starlings were subsequently reared in captivity and further tested during a simulated autumn migration. Migratory activity and orientation were tested using Emlen funnel trials over 6 consecutive weeks. Across treatment groups, we found a significant increase in mass, fat, and feather moult, and decreasing plasma thyroid hormones over time. At 12L:12D, control birds showed a peak in activity and a directional preference for 155.95° (South-southeast), while high-dosed birds did not. High-dosed birds showed a delayed directional preference for 197.48° (South-southwest) under 10L:14D, concomitant with apparent delays in moult. These findings link alterations in avian migratory behaviour to contaminant-specific mechanisms. Exposure to a ubiquitous environmental endocrine disruptor exerted only subtle short-term effects during the period of exposure but importantly, latent effects may be far more relevant for individual fitness. We discuss how the impacts of exposure during early stages of development were not significant for short-term nest success, but can still give rise to longer time-scale effects that are potentially relevant for survival and population stability for migratory birds.

Thesis (Ph. D.)--Michigan State University. Dept. of Zoology & Environmental Toxicology, 2008. / Title from PDF t.p. (viewed on Mar. 30, 2009) Includes bibliographical references. Also issued in print.

Thesis (M.S.)--University of Alabama at Birmingham, 2010. / Title from PDF t.p. (viewed June 30, 2010). Includes bibliographical references (p. 19-22).