scholarly journals Biological Activity of Wastewater Assessed Using in Vitro Cell-Based Assays

Author(s):  
Adamo R. Petosa ◽  
Monica Nowierski ◽  
Viviane Yargeau

Abstract Bioanalytical tools, namely in vitro bioassays, can be employed in tandem with chemical analyses to assess the efficacy of wastewater treatment and the potential for adverse effects from the discharges of wastewater into receiving waters. In the present study, samples of untreated wastewater (i.e. influent) and treated wastewater (i.e. effluent) were collected from two wastewater treatment plants and a wastewater treatment lagoon serving municipalities in southern Ontario, Canada. In addition, grab samples of surface water were collected downstream of the lagoon discharge. After solid phase extraction (SPE) using ion-exchange columns for basic/neutral and acidic compounds, respectively, the extracts were analyzed for a suite of 16 indicator compounds. The two SPE extracts were combined for analysis of biological responses in four in vitro cell-based bioassays. The concentrations of several indicator compounds, including the estrogens, 17β-estradiol and 17α-ethinylestradiol, were below the limits of detection. However, androstenedione and estrone were detected in several influent samples. The concentrations of these steroid hormones and some of the other indicator compounds declined during treatment but acesulfame K, carbamazepine, trimethoprim and DEET persisted in the effluent. The MTS- CellTiter 96® AQueous One Solution Cell Proliferation Assay (MTS) indicated that cell viability was not affected by exposure to the extracts. The Qiagen Nuclear Receptors 10-Pathway Reporter Array indicated that several cellular pathways were upregulated, with the greatest upregulation observed with the estrogen receptor (i.e. induction ratios 12 to 47) and the liver X receptor (i.e. induction ratios 10 to 45). The ERα CALUX assay indicated that estrogenic activity was lower in effluents compared to influents, with the greatest estrogenic activity observed for grab samples of influent from the lagoon (i.e. 56-215 ng L-1 17β-estradiol equivalents). Finally, the results of the Nrf2 Luciferase Luminescence Assay indicated a lower oxidative stress in the effluent samples. Overall, the present study demonstrates that chemical analyses are limited in their ability to predict or explain reductions in the toxicity of treated wastewater. There are thus advantages to using a combination of chemical analyses and in vitro bioassays to monitor the treatment efficiency of wastewater treatment plants and to predict the potential impacts of wastewater discharges into receiving waters.

2006 ◽  
Vol 54 (10) ◽  
pp. 87-93 ◽  
Author(s):  
T. Hashimoto ◽  
K. Takahashi ◽  
T. Murakami

Since the natural estrogens 17 β-estradiol (E2) and estron (E1), and the synthetic estrogen 17 α-ethynyl estradiol (EE2) have strong endocrine disrupting effects and the tendency to persist in effluent from wastewater treatment plants, effective measures are needed to remove them from wastewater. In this research, to gain an understanding of the characteristics of estrogen decomposition by ozonation, experiments were conducted using effluent from an actual wastewater treatment plant. In this experiment, estrogen was added to effluent at a concentration of 200 ng/l and 20 ng/l before the ozonation experiments. The results showed 90% or more of estrogen concentration and estrogenic activity of E2, E1 and EE2 to be removed at an ozone dose of 1 mg/l. At an ozone dose of 3 mg/l, the estrogen concentration and estrogenic activity of E2, E1 and EE2 in the treated water fell below the detection limit. The removal rate was not influenced by the kind of estrogen. No generation of byproducts with estrogenic activity was observed. The authors conclude that estrogen in secondary treated wastewater can be almost entirely removed at the practical ozone dose rate applied for the purpose of disinfection, which is up to about 5 mg/l.


2021 ◽  
Author(s):  
Semase Matseleng ◽  
Ozekeke Ogbeide ◽  
Patricks' Otomo Voua

Abstract Wastewater treatment facilities in developing countries like South Africa are major sources of contaminants via effluent into the environment, which could portend high toxicity risks for non-target flora and fauna. To this end, a study was conducted to determine the ecotoxicological responses of selected organism to treated and untreated wastewater from the wastewater treatment plants in an industrial town. The snail Helix pomatia was exposed to OECD artificial soil spiked with untreated or treated wastewater at the following concentrations: 0, 25, 50, 75, 100%. The ecotoxicological responses of Helix pomatia to wastewater were determined by assessing the biomass, survival, reproduction and biomarker responses (Catalase ‒ CAT and Acetylcholinesterase ‒ AChE activities). The overall results showed significant effects on the survival, reproduction and biomass of H. pomatia. Similar results were observed for juvenile emergence. An EC50 of 5.751% for egg production and an EC50 of 6.233% for juvenile emergence were determined in the untreated wastewater. Such indices could not be computed for the treated wastewater, indicating a decreased in toxicity between the untreated and the treated samples. For both the AChE and CAT activities, there was no statistical difference between treated and untreated wastewater treatments. The results from this study highlight the toxic effects of untreated wastewater and indicate that treated wastewater (effluent) released from the wastewater treatment plant in Phuthaditjhaba remains suitable for invertebrate fauna such as H. pomatia.


2011 ◽  
Vol 6 (5) ◽  
pp. 829-837 ◽  
Author(s):  
Mirjana Bistan ◽  
Romana Logar ◽  
Tatjana Tišler

AbstractEstrogenic activity has been detected in aquatic ecosystems across the world. However, there is a lack of such data for Slovenian wastewaters and surface waters. The Slovenian monitoring program of effluents discharged into surface waters does not require that emissions of natural and synthetic estrogens into aquatic environments be assessed and controlled. In our study, we assessed the potential estrogenicity of wastewater samples from three wastewater treatment plants using a yeast estrogen screen assay (YES assay). Due to the high inhibition of yeast growth in samples obtained during our first sampling period, it was impossible to detect any estrogenic activity. An additional silica gel clean-up step reduced the toxicity of samples collected during our second sampling period; as a result, we were able to record up to 95% relative estrogenic activity inhibition. Deconjugation of the estrogens did not significantly influence our results. We detected estrogenic activity using a YES assay in almost all influent and effluent samples tested, which suggests that the wastewater treatment plants (WWTPs) examined do not effectively remove (xeno)estrogens from wastewaters. Our results suggest that a YES assay is an appropriate screening method for monitoring estrogenic activity in effluents. However, prediction of the potential impacts of wastewater (xeno)estrogens on aquatic organisms require additional in vitro and in vivo assays.


2014 ◽  
Vol 931-932 ◽  
pp. 721-726 ◽  
Author(s):  
Apaporn Ruchiraset ◽  
Sopa Chinwetkitvanich

This study is the beginning of attempts to obtain existing data of estrogens contamination in wastewater treatment plants (WWTPs) in Bangkok Metropolitan area. Influents and effluents of seven WWTPs in Bangkok, and water samples from Bang-sue canal (receiving water from Chatuchak (CTC)-WWTP) and Chaopraya River (as receiving water from Chongnonsri (CNS)-WWTP) were collected by grab sampling. The sampling and analyses were done three times in a year round, during March to April 2010, October 2010, and January 2011. 17β-estradiol (E2) was found as in the majority in most samples including the surface water samples. In addition, the results of receiving water samples in both locations show that downstream (of discharged point) estrogens were a little higher than upstream ones. This could imply that WWTPs might be the important sources of estrogens contamination in surface (receiving) waters in Bangkok area and that the biological wastewater treatment processes currently used in WWTPs could not completely remove these estrogens.


2021 ◽  
Vol 5 (3) ◽  
pp. 125-130
Author(s):  
Natasha Berendonk Handam

Determining the presence of endocrine disrupting substances in waters is a relevant aspect for monitoring environmental health. Given its relevance, it is important to use methods that can make the total concentration of substances with estrogenic activity (eg endocrine disruptors), being faster, and without the use of compounds that pollute the environment. The purpose of the study was to compare the effectiveness of the methods of concentration by lyophilization and by vacuum concentration of substances with estrogenic activity present in reused water, using the commonly used methodology, solid phase extraction. Three methods were compared: solid phase extraction, lyophilization, and vacuum centrifugation. Sample aliquots of reused water received 17β-estradiol at a final concentration of 2 μg L-1 and were concentrated by the three methods. The analysis of estrogenic activity was performed by the in vitro YES (Yeast Estrogen Screen) assay. The results showed that the vacuum centrifugation, solid phase extraction and lyophilization methods had different percentages in the recovery of substances with estrogenic activity, being 45%, 40%, and 31%, respectively. The study pointed out that the lyophilization and vacuum centrifugation methods were effective as alternative methods for concentrating samples containing substances with estrogenic activity.


2013 ◽  
Vol 2013 ◽  
pp. 1-15 ◽  
Author(s):  
T. Vega-Morales ◽  
Z. Sosa-Ferrera ◽  
J. J. Santana-Rodríguez

Liquid and solid samples from two wastewater treatment plants (WWTPs) on Gran Canaria Island (Spain) have been tested for the presence of compounds with endocrine-disrupting properties. The selected degradation stages were sampled bimonthly from each WWTP over the 12-month period from July 2010 to July 2011. The analytical methods used for the determination of the endocrine-disrupting compounds (EDCs) were based on on-line solid phase extraction, microwave-assisted extraction (MAE), and ultrasonic-assisted extraction (UAE) coupled to UHPLC-MS/MS. All of the hyphenated methodologies employed in this work showed good recoveries (72–104%) and sensitivities, with LODs lower than 7.0 ng L−1and 6.3 ng g−1for the dissolved and solid fractions, respectively. We have also evaluated the estrogenicity of the samples in terms of their estradiol equivalent concentrations (EEQs). The chemical analysis of the selected EDCs revealed fairly low concentrations for both natural and synthetic oestrogens, alkylphenolic compounds, and bisphenol-A in each of the dissolved, particulate, and sludge samples (ng L−1or ng g−1). However, the estimated estrogenic activity indicated that the majority of samples could represent an important environmental risk, clearly surpassing the threshold to exert deleterious consequences on living beings.


Ecotoxicology ◽  
2017 ◽  
Vol 26 (10) ◽  
pp. 1327-1335 ◽  
Author(s):  
Yien Fang Ting ◽  
Sarva Mangala Praveena ◽  
Ahmad Zaharin Aris ◽  
Sharifah Norkhadijah Syed Ismail ◽  
Irniza Rasdi

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