The University of North Texas Stream Research Facility (UNTSRF) was designed to examine contaminant impacts on effluent-dominated stream ecosystems. Stream mesocosms, fed municipal effluent from the City of Denton, TX, Pecan Creek Water Reclamation Plant (PCWRP), were treated with 0, 15 or 140 µg/L cadmium for a 10-day study in August 2000. Laboratory toxicity test and stream macroinvertebrate responses indicated that cadmium bioavailability was reduced by constituents of effluent-dominated streams. The Biotic Ligand Model (BLM) for Cd was used to predict a 48 hour Cd EC50 for Ceriodaphnia dubia of 280 µg/L in these effluent-dominated streams. This value is higher that an EC50 of 38.3 µg/L Cd and a 7-day reproduction effect level of 3.3 µg/L Cd generated for C. dubia in reconstituted laboratory hard water. These results support use of a cadmium BLM for establishing site-specific acute water quality criteria in effluent-dominated streams. Although not affected by 15 µg/L treatments, organisms accumulated Cd in 15 µg/L treated streams. Hence, over longer exposure periods, Cd accumulation may increase and a no effect level may be lower than the observed 10-day no effect level of 15 µg/L. A toxicity identification evaluation procedure was utilized with in vitro and in vivo bioassays to identify estrogenic compounds in PCWRP effluent, previously identified to seasonally induce vitellogenin (VTG) in male fathead minnows. Steroids, nonylphenol ethoxylate metabolites, and other unidentified compounds were identified as causative effluent estrogens. These findings suggest that in vivo VTG bioassays should be used to confirm in vitro Yeast Estrogen Screening assay activity when effluents are fractionated or screened for estrogenicity. A subsequent 90-day cadmium study was initiated to assess long-term effluent and cadmium effects on fish endocrine function. Juvenile fathead minnows were placed in UNTSRF pool sections of replicate streams treated with 0, 5, 20 or 80 µg/L Cd. Male VTG was induced at each treatment level, indicating that PCWRP effluent was estrogenic during fall 2001. 20 and 80 µg/L Cd treatments reduced male circulating estradiol levels and critical swimming performance. Future studies are needed to assess impacts of environmental estrogen exposure on fish calcium metabolism and vertebral integrity.

The Pecan Creek Waste Reclamation Plant in Denton, Texas, an activated sludge WWTP, was sampled monthly for ten months to determine seasonal and site variation in concentrations of triclosan (5-chloro-2-(2,4-dichlorophenoxy)phenol), an antibacterial additive. SNK separation after the highly significant ANOVA on ranked data were: summer = fall > winter = spring and influent > downstream = effluent = wetland inflow > wetland outflow (a=0.05). After the plant converted to ultraviolet disinfection, measurements were made before and after the UV basin to determine if significant amounts of triclosan were converted to dioxin. Percent loss at each of the treatment steps was determined. Concentrations of triclosan in the downstream site were below the published NOEC for the most sensitive species.

Wetland wastewater treatment offers low-cost, energy efficient alternatives to conventional wastewater technologies. In this study, an artificial wetland was constructed at the City of Denton, Texas Pecan Creek Water Reclamation Plant to facilitate diazinon removal from treated effluent.

This study evaluated the effects of chlorinated effluent discharged from the City of Denton, Texas' wastewater treatment plant on Pecan Creek's fish and macroinvertebrate assemblages, and their recovery upon dechlorination. A baseline of ecological conditions was established while chlorine was present in the effluent (June 1993- October 1993), and was evaluated again after dechlorination with sulfur dioxide (October 1993-August 1994). In situ Asiatic clam and fathead minnow ambient toxicity tests, and fish and macroinvertebrate collections were used to establish this baseline for comparison to post-dechlorination results.

Dischargers to the Trinity River in North Texas were required to dechlorinate their effluents in 1990-91. Field surveys were conducted above and below an outfall to determine the response of resident immature insects and caged in situ juvenile Asiatic clams to chlorinated and dechlorinated effluent. Within six months after dechlorination began, insect community composition and C. fluminea survival significantly improved at stations below the outfall. Significantly lower clam growth within one mile below the dechlorinated effluent indicated the presence of non-chlorine toxicants. Effects from chlorinated and dechlorinated effluent exposure were comparable between Ceriodaphnia dubia lab tests and in situ C. fluminea.

Federal regulations such as the Clean Water Act (P.L. 92-500), and its amendments, direct the Environment Protection Agency (EPA) to implement programs to control the releases of conventional pollutants and toxics into the waterways of the United States. The EPA began requiring treatment plants to conduct toxicity tests (biomonitoring) of their effluent discharges. To control toxicity caused by chlorination of wastewater discharges, the EPA also began requiring some treatment facilities to dechlorinate their wastewater before discharging. This research was funded by the EPA to document the changes that occurred in the Trinity River from the dechlorination of the effluent from Ft. Worth's Village Creek municipal wastewater treatment plant. The study occurred over a two year period beginning in August 1990. A wide variety of biological field assessments and toxicological assays were used to measure various responses. Seven river stations, covering approximately twenty river miles, and the treatment plant effluent were assessed. Two of the river stations were upstream from the treatment plant and used as reference sites. The remaining five river stations were downstream from the treatment plant, spread out over seventeen river miles. The study evaluated the impact of chlorination prior to dechlorination, which served as a baseline. Responses determined during dechlorination were compared to the baseline data. An overall improvement in species richness and diversity was seen at those river stations which had previously been adversely impacted by chlorine. Aquatic toxicity tests, such as those required to be used by dischargers, were conducted during this study. Periodic toxicity was observed with these tests in the effluent and river samples after dechlorination was initiated. Those tests, along with in situ toxicity assays, proved to be good predictors of biological community responses.

The City of Denton Pecan Creek Wastewater Treatment Facility has periodically failed effluent toxicity testing. A Toxicity Identification Evaluation has determined that Diazinon in the effluent is contributing to the observed toxicity. Chlorpyrifos is also implicated as a factor. The City of Denton constructed a half acre experimental wetland to remove Diazinon related toxicity. Results from spiking and microcosm experiments indicate that the wetland can reduce the Diazinon.

This investigation had for its purpose the determination of first, the inefficiency of the Perryton, Texas, sewage disposal plant proper; and second the potential effectiveness of a lagoon now used to complete the sewage purification; and consequently, offset an apparent health hazard. The problem, as pursued, has consisted of first, the collection of sewage samples from the six stations listed in Table 1 over the period of time from September 23, 1949 to May 14, 1950; second, a quantitative analysis of each sample in order to determine the number of bacterial organisms present; third, a qualitative analysis of each sample in order to determine more expecially the presence of members of the genera Aerobacter, Escherischia, Salmonella and Shigella; and fourth, an attempt to evaluate the potentialities of the natural purification factors of the lagoon.

MSc (Microbiology) / Department of Microbiology / Background: The occurrence of microorganisms and endocrine disrupting chemicals (EDCs) in water poses a serious concern due to their effects on humans, animals and environment. In recent years, EDCs have been increasingly reported in rivers that receive large amounts of wastewater effluents. Of all the EDCs, natural and synthetic hormones are among those that are recognized for their potential to mimic or interfere with normal hormonal functions of humans and animals. The present study aimed at assessing the occurrence of these hormones in relation to the molecular diversity of Aeromonas and evaluating the resistance of Aeromonas to antibiotics as well as to assess anti-bacterial activity of two selected traditional medicinal plants. Methods: Wastewater, water and fish samples were collected from various sources (rivers, wastewater treatment plants, taps, and dams) for the detection of hormones and isolation of Aeromonas species. The analysis of hormones from various organs of the fish and from water samples was conducted, after extraction using enzymelinked immunosorbent assays (ELISA). Different types of hormones including Estriol, Estradiol, Ethinylesradiol and Testosterone were detected, and their concentrations determined. Aeromonas spp were isolated rom the samples using microbiological methods and Conventional PCR was used for genotyping as well as for detection of the beta-lactamase genes. Kirby-bauer method was used to determine the susceptibility profiles of Aeromonas to different antibiotics. Microdilution assay was used to determine the Anti-bacterial activity of the plant (Annoniceae and Zornia milneana) extracts against Aeromonas species. Results: A total of 144 samples were collected from 23 different locations in two countries: South Africa and Zambia. These included wastewater and treated wastewater, River water, fish and tap water. 17α-ethinylestradiol (EE2) was detected in most of the samples (92.7%) with concentrations varying from 0.59 ng/ml to 65 ng/ml. The hormones were also detected from drinking water, with testosterone detected at high concentrations of up to 140 ng/ml in tap water. Most sewage treatment plants were not able to remove the EE2 from the wastewater as the concentration of this hormone in the final effluent was almost always higher than that in the influent. These homones were also detected in drinking water at high concentrations of up to 53.49 ng/ml in the tap water for EE2 and 1777 ng/ml for E2. The overall detection of Aeromonas species in the samples was 84.5%. A. caviae was the most prevalent species accounting for 73.6%, followed by A. veronii with 64.6%. The bacteria were completely resistant to cefuroxime accounting for 100% resistance. Aeromonas isolates also showed high resistance to trimethroprim (88.7% for A. hydrophila), cefazolin (highest 97.8% for A. cavie), and ceftazidime (83.9% for A. sobria). TEM was the most prevalent beta-lactamase gene with detection rate of 87%. All isolates lacked the presence of the CTX-M3 gene. Also, wastewater had the highest prevalence of A. veronni and A. caviae accounting for 87.5% and 82.5% respectively. Multiple antibiotic resistance was also observed with the Aeromonas isolates being resistant to up to 11 antibiotics. High prevalence of 77.1% of Aeromonas hydrophila was observed in the presence of ethinylestradiol (EE2). Aeromonas veronii and Aeromonas caviae were the most predominant species in the presence of total estriol, A. veronii had a prevalence of 57.1% and A. caviae had a prevalence of 52.8%. Aeromonas hydrophila and Aeromonas caviae had the lower prevalence in the presence of hormones with the percentages of 26.1% and 27.8% respectively. The methanol extracts of both Zornia milneana and Annona species showed good activity against the Aeromonas spp with the lowest MIC of 0.078 mg/ml. Ethyl acetate extracts were the least effective. Conclusion: This study has shown high occurrence of steroid hormones in all types of environmental samples tested. These included tap water, river water, wastewater and fish both in Zambia and South Africa. Therefore, steroid hormones constitute and important health problem in the Southern African Sub-Region. The incapacity of the wastewater treatment plants to remove EE2 is an important problem that needs to be tackled immediately. The prevalence of Aeromonas species is very high in our environmental water as well as in drinking water, with the highest prevalence observed in fish and wastewater. It was also revealed that there is relationship between steroid hormones and Aeromonas species, with the hormones supporting the growth of Aeromonas species. The presence of beta-lactamase genes which causes Aeromonas to be resistant to antibiotics was also noted. Methanol extracts of Zornia milneana and Annona spp were the most effective against Aeromonas spp and could serve as primary sources for the isolation of lead compounds. / NRF