Evaluation of an Exposure Assay to Measure Uptake of Sediment Pah by Fish

The ecological risks of polynuclear aromatic hydrocarbons (PAH) in aquatic sediments will vary with both toxicity and bioavailability to aquatic biota. While there are standardized protocols to test the acute toxicity of sediment-borne compounds to aquatic invertebrates, there are none for assessing bioavailability to fish. We found that sediment-borne PAH caused an exposure-dependent induction of cytochrome P450 (CYP1A) enzymes in rainbow trout (Oncorhynchus mykiss) fingerlings exposed in semi-static 96 h bioassays, as shown by increased activity of ethoxyresorufin-o-deethylase (EROD). Assuming that PAH are taken up by trout due to partitioning from organic and inorganic constituents of sediments, we tested the effect of different test variables on bioaccumulation using induction as an index of exposure. EROD activity increased with exposure of fish to increasing volumes of sediments containing PAH, i.e., with increasing ratios of sediment to water. Uptake of single compounds from sandy sediments did not differ from uptake from clay or low organic (7% LOI — loss on ignition) sediments, but decreased when organic content was very high (58% LOI). Maximum induction was observed within 24~h of exposure and at 7.5^C relative to 15, 22, or 28^C. Storage and handling techniques had minor effects on sediment EROD induction potency. Absolute EROD activity was greater in white sucker (Catostomus commersoni) a benthic species, than in trout, a pelagic species. However, when basal (negative control) activity was accounted for, there was no difference in response between the species. Together, these experiments provide a basis for developing a standard protocol to test the bioavailability to fish of sediment-borne PAH.     

Determination of atrazine in surface waters by combination of POCIS passive sampling and ELISA detection

Polar organic compound integrative samplers (POCIS) in combination with instrumental techniques such as LC-MS-MS were previously used to monitor environmental pollutants but the performance of alternative immunochemical methods such as ELISA (enzyme-linked immunosorbent assay) has been explored less. In the present study, POCIS technology was applied to surface water sampling in the Czech Republic, and ELISA was used as a detection technique for the herbicide atrazine. In the first study, 28 samples from streams around small municipal waste water treatment plants (WWTPs) were collected using two different devices (POCISpest and POCISpharm) over the course of 21 days. Elevated atrazine concentrations (up to 25 ng per POCIS) were found in samples down-stream of WWTPs. This observation was also confirmed in another two year study (4 sampling periods) investigating 7 river sites around a major city of Brno as well as the inlet and outlet of the city's WWTP. High atrazine levels were systematically determined at the outlet from the WWTPs (120-605 ng per POCIS). A decreasing trend in the atrazine concentrations in rivers around the city of Brno has been observed, with the highest levels observed within the first sampling period in spring 2007 (100-600 ng per POCIS, with an extreme value of 2760 ng per POCIS). Results of the atrazine ELISA were closely correlated with LC-MS/MS, which confirmed good applicability of ELISA as a cost-effective screening tool.     

中国表层水体沉积物中多环芳烃源解析及评价

采用索氏提取气相色谱-质谱法测定中国6个重点水体表层沉积物中16种多环芳烃的含量。各化合物含量范围分别为长江6.20~163 ng/g、淮河7.90~249 ng/g、海河12.1~401 ng/g、松花江5.75~152 ng/g、太湖29.1~2 810 ng/g和滇池19.1~795ng/g；16种多环芳烃的总量分别为：长江1 147 ng/g、淮河1 723 ng/g、海河2 595 ng/g、松花江793 ng/g、太湖12472 ng/g、滇池3 714 ng/g，属中等污染水平。利用特征分子比值法分析结果表明6条水体表层沉积物中PAHs均可能以燃料（包括柴油、汽油、煤、木材）燃烧以及焦化污染为主。淮河和滇池还可能存在轻微石油泄漏污染。利用沉积物质量基准法（SQGs）和沉积物质量标准法分别对6条水体表层沉积物中多环芳烃的风险评估表明严重的多环芳烃生态风险在这些水体表层沉积物中不存在，但长江、淮河、松花江、海河均可能存在一定的潜在风险，负面生物毒性效应会偶尔发生，风险主要来源于荧蒽和菲。太湖和滇池水体中存在的潜在多环芳烃风险种类较多，风险主要来源于菲、荧蒽、芘、苯并（a）蒽、苊和蒽，对水生生物毒性效应较高，有必要进行更深入细致的调查研究高风险区域底栖生物的受损状况、污染来源和途径，以制定合理的污染控制对策。     

Composition and source apportionment of PAHs in sediments at river mouths and channel in Kaohsiung Harbor, Taiwan

Fifty-eight sediment samples were collected in 2009 from the bottom of river mouths near Kaohsiung Harbor (Taiwan) and the harbor channel for the analyses of polycyclic aromatic hydrocarbons (PAHs) using gas chromatography-mass spectrometry (GC-MS). Concentrations of total PAHs varied from 39 to 30,521 ng g(-1) (dry weight); samples collected from the mouths of Love River, Canon River, Jen-Gen River, and Salt River showed the highest PAHs concentrations. This indicates that the major sources of sediment PAHs come from those polluted urban rivers and the harbor channel. In samples collected from the Salt River mouth, approximately 43% of the PAHs are identified as PAHs with 2 or 3 rings. However, samples collected from other locations contain predominantly PAHs with 4 rings (32 to 42%) or 5 and 6 rings (36 to 44%). Emissions from traffic-related sources and waste incineration contribute to the majority of PAHs found in most channel and river mouth sediments. However, coal/oil combustion is the main cause of high concentrations of PAHs observed in the Salt River mouth sediments. Principal component analyses with multivariate linear regression (PCA/MLR) have been used to further quantify the source contributions, and the results show that the contributions of coal/oil combustion, traffic-related and waste incineration are 37%, 33% and 30%, respectively.     

An examination of the toxic properties of water extracts in the vicinity of an oil sand extraction site

The industrial extraction of oil sands (OS) in northern Alberta, Canada, has raised concerns about the quality of the Athabasca River. The purpose of this study was to examine the toxic properties of various water extracts on Oncorhynchus mykiss trout hepatocytes. The water samples were fractionated on a reverse-phase C(18) cartridge and the levels of light-, medium- and heavy-weight polycyclic aromatic hydrocarbons (PAHs) were determined by fluorescence spectroscopy. Primary cultures of trout hepatocytes were exposed for 48 h at 15 °C to increasing concentrations of the C(18) extract corresponding to 0.02, 0.1, 0.5 and 2.5X concentrations from upstream/downstream sites in the Athabasca River, lake and groundwater samples, OS tailings and interceptor well-water samples. Changes in cell viability, phase I and phase II biotransformation enzymes (cytochrome P4501A and glutathione S-transferase activities), oxidative damage (lipid peroxidation LPO) and genotoxicity (single and double DNA strand breaks) were monitored in post-exposure cells. The water samples decreased cell viability and increased all the above endpoints at thresholds of between 0.02 and 0.1X the water concentration. The most responsive biomarker was DNA damage but it also offered the least discrimination among sites. LPO was higher at sites downstream of the industrial operations compared to upstream sites. A decision tree analysis was performed to formulate a set of rules by which to identify the distinctive properties of each type of water samples. The analysis revealed that OS tailings and interceptor waters were characterized by an increased concentration in light PAHs (>42 μg L(-1)) and this fraction represented more than 85% of the total PAHs. These samples also inhibited GST activity, which could compromise the elimination of genotoxic PAHs present in the system. An analysis of groundwater samples revealed a contamination pattern similar to that for OS tailings. There is a need for more research into specific biomarkers of toxicity from OS tailings compounds such as naphthenic acids, light PAHs among others, which are a characteristic fingerprint of OS extraction activities.     

Distribution and sources of polycyclic aromatic hydrocarbons in Wuhan section of the Yangtze River, China

Polycyclic aromatic hydrocarbons (PAHs) are important organic contaminants with great significance for China, where coal burning is the main source of energy. In this study, concentrations, distribution between different phases, possible sources and eco-toxicological effect of PAHs of the Yangtze River were assessed. PAHs in water, suspended particulate matters (SPM) and sediment samples at seven main river sites, 23 tributary and lake sites of the Yangtze River at the Wuhan section were analyzed. The total concentrations of PAHs in the studied area ranged from 0.242 to 6.235 μg/l in waters and from 31 to 4,812 μg/kg in sediment. The average concentration of PAHs in SPM was 4,677 μg/kg, higher than that in sediment. Benzo(a)pyrene was detected only at two stations, but the concentrations were above drinking water standard. The PAHs level of the Yangtze River was similar to that of some other rivers in China but higher than some rivers in foreign countries. There existed a positive relationship between PAHs concentrations and the TOC contents in sediment. The ratio of specific PAHs indicated that PAHs mainly came from combustion process, such as coal and wood burning. PAHs may cause potential toxic effect but will not cause acute biological effects in sedimentary environment of the Wuhan section of the Yangtze River.     

Earthworm cytochrome P450 determination and application as a biomarker for diagnosing PAH exposure

We developed a new microsome purification method and used the Omura and Sato method to measure the total content of cytochrome (Cyt) P450 in earthworm (Eisenia fetida) microsomes. In method development, two different pretreatments, i.e. solubilization or manual separation were used to purify worm microsomes. Solubilization was more effective than manual separation and difference spectra showed a peak at 450 +/- 1 nm in microsomes received solubilization pretreatment. We conducted a 48 h contact test by exposing worms on pyrene (Py) and benzo[a]pyrene (BaP) spiked filter paper. A dose-response relationship was established between total P450 content and the concentration of Py or BaP ranging from 10(-6) mg mL(-1) to 10(-2) mg mL(-1). Results show that total Cyt P450 content in earthworms is a promising biomarker for diagnosing PAHs exposure at sublethal dose ranges.     

Kinetic of Biomarker Responses in Juveniles of the Fish Sparus aurata Exposed to Contaminated Sediments

Sediments in the National Park of the Atlantic Islands (Galicia, Spain) were affected by the spill of the tanker Prestige (November, 2002) and still present high levels of Polycyclic aromatic hydrocarbons. The adverse effects associated with the contaminants in sediments were tested using a chronic bioassay, exposing juveniles of the fish Sparus aurata (seabream). A toxicokinetic approach is proposed to evaluate sediment quality by linking chemical and ecotoxicological data along the time. Sediment samples were physicochemically characterized and the concentration of contaminants (Polycyclic aromatic hydrocarbons – PAHs – and metals) was measured. Fishes were exposed to contaminated sediments, and samples from different tissues were collected every 15 days throughout the 60 days that lasted the experiment. A biomarker of exposure (ethoxyresorufin O-deethylase activity – EROD activity) and a biomarker of effect (histopathology) were analyzed during the exposure period. Results show a relationship between the biomarkers and the concentrations in sediments of polycyclic aromatic hydrocarbons—PAHs. Besides, the toxicokinetic approach links biomarkers response providing information about the relationship between the detoxification process and the damages observed in the different tissues. The frequency of the histological damage is highest when the EROD activity slightly decreases in accordance with the mechanism of detoxification of this enzymatic system against PAHs and other organic contaminants.     

The responses of the hepatosomatic index (HSI), 7-ethoxyresorufin-O-deethylase (EROD) activity and glutathione-S-transferase (GST) activity in sea bass (Dicentrarchus labrax, Linnaeus 1758) caged at a polluted site: implications for their use in environmental risk assessment

The present study investigates the response of three hepatic biomarkers in adult sea bass (Dicentrarchus labrax, Linnaeus 1758) caged at a wastewater outlet of an oil refinery with fish caged at a pristine site used as controls. The biomarkers that were investigated were the hepatosomatic index (HSI), 7-ethoxyresorufin-O-deethylase (EROD) activity and glutathione-S-transferase (GST) activity. In addition, we have measured the levels of polycyclic aromatic hydrocarbons (PAHs) and selected heavy metals (lead, cadmium, mercury, copper and zinc) in sediment samples at the polluted site. Although the polluted site had high environmental levels of PAHs and heavy metals, there was no difference in hepatic EROD activity and HSI between fish caged at the polluted site and controls. On the other hand, GST activity was significantly lower in fish caged at the polluted site compared to controls. Our results point out that the studied biomarkers have limited use in environmental risk assessment studies, at least when caged adult sea bass is used as the sentinel species and complex toxicant mixtures are involved.     

Measurement of organic contaminants and biological effects in Scottish waters between 1999 and 2005

Concentrations of organic contaminants were determined in sediment and fish collected annually at six sites around Scotland between 1999 and 2005, as part of the UK National Marine Monitoring Programme. Polycyclic aromatic hydrocarbons (PAHs) and chlorobiphenyls (CBs) were measured in sediment, while CBs and ethoxyresorufin-O-deethylase (EROD) activities were measured in the fish. Highest PAH concentrations in sediment were found at sites where higher coastal influences would be expected (e.g., Clyde and Solway) and four of the six sites gave PAH concentrations above OSPAR background assessment concentrations (BACs). A significant downward trend in the median total PAH concentrations, normalised to total organic carbon (TOC), was found at one of the six sites (Minches). The PAH profiles at all sites were consistent over the 6 year period, indicating that the sites are relatively stable and PAH sources are not changing. There was an indication of a greater petrogenic input on the west coast, with sediment from the Clyde, Solway and Minches having a greater proportion of 2- and 3-ring PAHs and a lower proportion of 5-ring PAHs. CB concentrations at all sites were low compared with UK estuarine sites and similar to sediment from more remote areas: however, BACs were exceeded. No significant trends were detected in either the sediment CB concentrations or patterns at any of the six sites. CB concentrations were significantly higher in fish liver collected from the Clyde. Concentrations at the other five sites were low, with the majority of samples having concentrations for the ICES7 CBs of <500 microg kg(-1) lipid weight. However, individual CB concentrations were still above the BACs. Hepatic EROD activities were measured in male plaice from 2002-2005 and were generally low (<10 pmol min(-1) mg protein(-1)). No temporal trends were detected in either CB concentrations or the EROD activity.     

Use of biomarkers in Nile tilapia (Oreochromis niloticus) to assess the impacts of pollution in Bolgoda Lake, an urban water body in Sri Lanka

The present study reports the first analysis of water pollutants in Sri Lankan waters using a suite of biomarkers in Nile tilapia (Oreochromis niloticus) residing in Bolgoda Lake which receives urban, industrial and domestic wastes from multiple sources. The fish were collected from the lake in the dry period (April 2005) and wet periods (September 2005, October 2006) and the levels of biomarkers viz. hepatic ethoxyresorufin O-deethylase (EROD), glutathione S-transferase (GST), metallothioneins, biliary fluorescent aromatic compounds, brain and muscle cholinesterases (ChE) were compared with those of the laboratory reared control fish and the fish obtained from a less polluted water body, Bathalagoda reservoir (reference site). The results revealed that biomarker levels of the fish collected from the reference site were not significantly different from the controls. Hepatic EROD and GST activities in fish from Bolgoda Lake were induced 4.2-16.6 folds and 1.4-3.3 folds respectively compared with the control fish. Analysis of bile in the lake fish revealed recent uptake of naphthalene, pyrene and benzo(a)pyrene type polycyclic aromatic hydrocarbons (PAHs). The induction of EROD activities in feral fish reflects the exposure of fish to aryl hydrocarbon receptor agonists including PAHs present as pollutants in the Bolgoda Lake. Cholinesterase activity in the fish inhabiting one sampling site of Bolgoda Lake was lower (22-40% inhibition) than the activity measured in the control fish indicating the presence of anticholinesterase pollutants in the area. Hepatic metallothionein levels in the lake fish were higher (1.9-3.2 folds) in comparison to the controls indicating metal exposure. The results support the potential use of these biomarkers in Nile tilapia in assessing pollution in tropical water bodies.     

Assessing the impacts of water abstractions on river ecosystem services: an eco-hydraulic modelling approach

The provision of important river ecosystem services (ES) is dependent on the flow regime. This requires methods to assess the impacts on ES caused by interventions on rivers that affect flow regime, such as water abstractions. This study proposes a method to i) quantify the provision of a set of river ES, ii) simulate the effects of water abstraction alternatives that differ in location and abstracted flow, and iii) assess the impact of water abstraction alternatives on the selected ES. The method is based on river modelling science, and integrates spatially distributed hydrological, hydraulic and habitat models at different spatial and temporal scales. The method is applied to the hydropeaked upper Noce River (Northern Italy), which is regulated by hydropower operations. We selected locally relevant river ES: habitat suitability for the adult marble trout, white-water rafting suitability, hydroelectricity production from run-of-river (RoR) plants. Our results quantify the seasonality of river ES response variables and their intrinsic non-linearity, which explains why the same abstracted flow can produce different effects on trout habitat and rafting suitability depending on the morphology of the abstracted reach. An economic valuation of the examined river ES suggests that incomes from RoR hydropower plants are of comparable magnitude to touristic revenue losses related to the decrease in rafting suitability.     

Cytochrome P450 system as potential biomarkers of certain toxicants: comparison between plant and animal models

In present study, we measured the activities of some selected cytochrome P450 isozymes like ethoxyresorufin-o-deethylase (EROD), pentoxyresorufin-o-deethylase (PROD), and N-nitrosodimethylamine demethylase (NDMA-d) treating experimental rats with different environmental toxicants, namely trichloroethylene (TCE), heavy metal mixture, and ethyl alcohol individually or in combination and by exposing the Allium cepa bulbs to increasing concentration of TCE for 48 h. In animal system, NDMA-d activity displayed a remarkable amplification by 4.2-fold in the liver of alcohol ingested rats compared with control animals. TCE intake also resulted in a marked increase of around 3.8-fold and 1.2-fold in rats’ NDMA-d activity in the liver and kidney, respectively. In onion bulbs, an amazing rise in the activity of test enzymes was recorded in a dose-dependent manner. Among the said enzymes, PROD showed maximum increase up to the extent of 22-fold in comparison with control at 20 ppm of TCE exposure, whereas NDMA-d and EROD showed 11- and 9-fold increase in enzymatic activity, respectively, compared with the control. Based on this study, we conclude and advocate that all the selected isozymes of CYP450 system, viz. PROD, NDMA-d, and EROD can act as potent biomarkers in plant system for assessing the TCE pollution.     

Biomonitoring with Gammarus pulex at the Meuse (NL), Aller (GER) and Rhine (F) rivers with the online Multispecies Freshwater Biomonitor

Biological early warning systems represent a set of tools that may be able to respond to certain chemical monitoring requirements of recent European legislation, the Water Framework Directive (WFD2000/60/EC), that aims to improve and protect water quality across Europe. In situ biomonitoring was performed along the rivers Meuse (NL), Aller (GER) and Rhine (F) within the frame of the European Union-funded Project SWIFT-WFD. Gammarus pulex was used as a test organism during the evaluation of the Multispecies Freshwater Biomonitor(R) (MFB), an online biomonitor to quantitatively record different behaviour patterns of animals. At the river Meuse G. pulex reacted to pulse exposure of either a mixture of trace metals or of several organic xenobiotics, by showing up to 20% decreased locomotory activity (already at the 1st pulse) and increased mortality (at 2nd or 3rd pulse only). G. pulex deployed within the MFB system were observed to survive well at the monitoring station on the Aller (100%) and monitoring did not result in the measurement of chemical irregularities. In contrast, deployment at the monitoring station on the Rhine river demonstrated that the test organism was able to detect chemical irregularities by up to 20% decreased locomotory activity in the animals. The MFB proved to be an alert system for water quality monitoring at sensitive sites and sites with accidental pollution.     

Comparison of general water quality of rivers in Indonesia and Japan

In Java and Kalimantan in Indonesia, river water plays important roles in human life; for example, for transportation, and economic activities of the inhabitants. However, industrial, agricultural and domestic water is discarded into rivers directly in many developing countries, including Indonesia, since drainage systems have not been completely constructed. In this study, to evaluate the water quality and to compare those levels of environmental contaminants in developing and developed countries, water quality and contents of endocrine disrupters were measured in a total of 64 water samples (Indonesia; 28 samples and Japan; 36 samples) from 53 sites. The results indicated that, rivers in both capital cities, Jakarta and Tokyo, were contaminated. Water in rivers in Indonesia was not so heavily polluted as in Japan. Pollution in the river water in Indonesia appeared to be caused by the lack of sewerage systems. In addition, the findings on endocrine disrupters indicated that the concentration of alkylphenol in water samples was large enough to affect living organisms.     

Polycyclic aromatic hydrocarbons in river sediments from the western and southern catchments of the Bohai Sea, China: toxicity assessment and source identification

The concentration, distribution, and origin of 16 USEPA priority polycyclic aromatic hydrocarbons (PAHs) were investigated in river sediments from the western and southern catchments of the Bohai Sea, China. A toxicity assessment of 28 sediment samples collected from 15 main rivers was conducted by utilizing threshold and probable effect concentrations (TEC and PEC, respectively) derived from consensus-based sediment quality guidelines. The concentrations of total PAHs (∑PAH₁₆) ranged from 0.14 to 10,757 μg/kg dry weight (mean?=?1,368.6 μg/kg). The high concentrations of PAHs found in Binhai New Area of Tianjin and Binzhou City are likely the result of rapid population and heavy industry growth, resulting in elevated point and nonpoint source inputs of PAHs. Of the samples collected, samples 1 and 10 (7.1 % of the total) were categorized as toxic since some of the PAH concentrations exceeded the corresponding PECs. Twenty samples were classified as nontoxic, with both the individual PAH and the∑PAH₁₆ concentrations falling below the corresponding TECs. Analyses of selected PAH isomer ratios aided in the identification of PAH origins, allowing for a discrimination between pyrogenic and petrogenic sources. Spatial variability confirmed source heterogeneity within the study area. The most significant PAH-associated contamination was found in the Beitang River and Ji Canal, which are located in Binhai New Area. The magnitude of contamination and the likelihood of an ongoing influx of PAHs support the need for a better understanding of pollution sources and methods for both control and sediment restoration.     

Polycyclic aromatic hydrocarbons in water, sediment and soil of the Songhua River Basin, China

The Songhua River is the third largest river in China and the primary source of drinking and irrigation water for northeastern China. The distribution of 16 priority polycyclic aromatic hydrocarbons (PAHs) in water [dissolved water (DW) and suspended particulate matter (SPM)], sediment, and soil in the river basin was investigated, and the associated risk of cancer from these PAHs was also assessed. The total concentration of PAHs ranged from 13.9 to 161 ng L^?1 in DW, 9.21 to 83.1 ng L^?1 in SPM, 20.5 to 632 ng g^?1 dw (dry weight) in sediment, and from 30.1 to 870 ng g^?1 dw in soil. The compositional pattern of PAHs indicated that three-ring PAHs were predominant in DW and SPM samples, while four-ring PAHs dominated in sediment and soil samples. The spatial distribution of PAHs revealed some site-specific sources along the river, with principal component analysis indicating that these were from pyrogenic sources (such as coal and biomass combustion, and vehicle emissions) and coke oven emission distinguished as the main source of PAHs in the Songhua River Basin. Based on the ingestion of PAH-contaminated drinking water from the Songhua River, cancer risk was quantitatively estimated by combining the Incremental Lifetime Cancer Risk assessment model and BaP-equivalent concentration for five age groups of people (adults, teenagers, children, toddlers, and infants). Overall, the results suggest that the estimated integrated lifetime cancer risk for all groups was in acceptable levels. This study is the first attempt to provide information on the cancer risk of PAHs in drinking water from the Songhua River.     

Determination of 7-ethoxyresorufin-o-deethylase (EROD) induction in leaping mullet (Liza saliens) from the highly contaminated Aliaga Bay, Turkey

Pollution of the aquatic environment is a global concern owing to the devastating effects of contaminants whose levels are growing at an alarming rate, and it has become a major threat for marine organisms, as well as to humans as consumers. This study has been carried out on leaping mullet obtained from Aliaga Bay, which is located on the west coast of the Aegean Sea near Izmir and hosts the world’s fifth largest shipyard, plus a broad range of industrial activities, including an oil refinery and a paper factory. The waste from these industries, combined with municipal sewer discharges, is the main cause of pollution in this region. There is no national documentation or research on the determination of pollution resulting from the industrial activities in this area. In the present study, the degree of induction of CYP4501A-associated 7-ethoxyresorufin O-deethylase (EROD) activity and immunochemical detection of CYP1A1 in the liver of leaping mullet (Lisa saliens) were used as biomarkers for the assessment of polycyclic aromatic hydrocarbon (PAH)-type organic pollutants in Aliaga Bay. Mullet caught from different locations of the bay had approximately 52 times more EROD activity than the feral fish sampled from a clean reference site near Foca, Izmir. The results of this study indicate that Aliaga Bay is highly contaminated with PAH-type organic pollutants.     

Induction of cytochrome P4501A and endocrine disrupting effects of school incinerator residues

The emission of the dioxin-like compounds from on-site waste incinerators of seven schools in Kyonggi Province of Korea was evaluated by determination of the cytochrome P4501A(CYP1A) catalytic activity and antiestrogenic activity using cell culturemicrobioassay. The residue samples were extracted in a Soxhlet apparatus using toluene for 20 hr. The concentrated crude extractswere fractionated with a basic alumina column. Dioxin-like compounds were then extracted. Induction of CYP1A activity in a rat(H4IIE) hepatoma cell line was used as indicator of biologicaleffect of incinerator residues and measured as 7-ethoxyresorufin-O-deethylase(EROD) activities. The EROD activities of fraction II extracts (one of the two extracts) in the H4IIE cells were from 0.044±0.002 to 4.424±0.351 ng-TEQ g^-1 (TCDD Toxicity equivalent), showing relatively high inducing capacity. Antisetrogenicity of the extracts was measured as decrease in E₂-induced cell proliferation. Most of the extracts showed antiestrogenic activity in MCF7-BUS cell.The TEQ levels of the incinerator residues and the antiestrogenicactivities were in good correlation, strongly suggesting that thepotent toxic emissions were indeed produced from the on-site school waste semi-incinerators and could cause the antiestrogenicity.