超声波降解四环素类抗生素废水

采用超声波降解模拟废水中四环素类抗生素(TCs):土霉素(OTC)、四环素(TC)和金霉素(CTC),考查废水初始浓度、初始pH、超声波输入功率密度、曝气量和自由基清除剂(正丁醇)对降解效率的影响。结果表明,超声波可以快速高效地降解废水中的TCs,CTC最易被降解,TC和OTC次之;在OTC、TC和CTC初始浓度0.25 mg/L,初始pH=8.2,声功率密度1.2 W/mL,气水体积比30∶1的条件下,超声波辐照20 min后去除率分别达到76.8%、84.0%和94.4%。在实验研究条件下,TCs去除率均随初始浓度(0.25~2.00 mg/L)的增大而降低,但总去除浓度升高;碱性条件有利于TCs降解,去除率随初始pH(6~11)升高而升高;功率密度(0.24~1.96 W/mL)越高,TCs去除率也越高;曝气后TCs的去除率明显升高,随着气水比(10∶1~60∶1)的增大,去除率逐渐上升;正丁醇有效抑制TCs超声波降解,可推断TCs主要降解途径为自由基氧化。TCs超声波降解过程符合伪一级动力学反应特征。     

水中四环素类化合物在不同光源下的光降解

研究了模拟太阳光和实际室外太阳光下四环素类化合物在水溶液中的光降解。结果表明,四环素类化合物(TCs)在模拟太阳光照射下均能发生直接光降解,且其降解速率随pH不同变化很大。四环素(TC)、土霉素(OTC)、金霉素(CTC)和多西环素(DTC)的光降解速率随溶液pH升高明显加快,而米诺环素(MTC)在水中的光降解速率随溶液pH升高略有下降。天然水体pH范围内,模拟太阳光照下四环素类化合物表观光降解速率常数在0.004～0.026 min-1范围内,降解半衰期在26～136 min范围内。在室外实际太阳光照下,四环素类化合物光降解反应速率与太阳光强成正比例关系。晴朗无云天气下,四环素类化合物降解半衰期在春冬季节较长、夏季节较短;四环素在东江实际水体中表观光降解速率很快,在冬季晴天天气下,其平均半衰期仅为8.2 min。     

Transformation kinetics and pathways of tetracycline antibiotics with manganese oxide

Tetracycline antibiotics including tetracycline (TTC), oxytetracycline (OTC) and chlorotetracycline (CTC) undergo rapid transformation to yield various products in the presence of MnO₂ at mild conditions (pH 4-9 and 22 °C). Reaction rates follow the trend of CTC > TTC > OTC, and are affected by pH and complexation of TCs with Mg²⁺ or Ca²⁺. Experimental results of TTC indicate that MnO₂ promotes isomerization at the C ring to form iso-TTC and oxidizes the phenolic-diketone and tricarbonylamide groups, leading to insertion of up to 2 O most likely at the C9 and C2 positions. In contrast, reactions of OTC with MnO₂ generate little iso-OTC, but occur mainly at the A ring’s dimethylamine group to yield N-demethylated products. CTC yields the most complicated products upon reactions with MnO₂, encompassing transformation patterns observed with both TTC and OTC. The identified product structures suggest lower antibacterial activity than that of the parent tetracyclines.     

Occurrence and distribution of heavy metals and tetracyclines in agricultural soils after typical land use change in east China

Land use in east China tends to change from paddy rice to vegetables or other high-value cash crops, resulting in high input rates of organic manures and increased risk of contamination with both heavy metals (HMs) and antibiotics. This investigation was conducted to determine the accumulation, distribution and risks of HMs and tetracyclines (TCs) in surface soils and profiles receiving different amounts of farmyard manure. Soil samples collected from suburbs of Hangzhou city, Zhejiang province were introduced to represent three types of land use change from paddy rice to asparagus production, vineyards and field mustard cultivation, and divided into two portions, one of which was air-dried and sieved through 2-, 0.3- and 0.149-mm nylon mesh for determination of pH and heavy metals. The other portion was frozen at ?20 °C, freeze-dried and sieved through a 0.3-mm nylon mesh for tetracyline determination. HM and TC concentrations in surface soils of 14-year-old mustard fields were the highest with total Cu, Zn, Cd and ∑TCs of 50.5, 196, 1.03 mg?kg^?1 and 22.9 μg?kg^?1, respectively, on average. The total Cu sequence was field mustard?>?vineyards?>?asparagus when duration of land use change was considered; oxytetracycline (OTC) and doxycycline were dominant in soils used for asparagus production; OTC was dominant in vineyards and chlortetracycline (CTC) was dominant in mustard soils. There were positive pollution relationships among Cu, Zn and ∑TCs, especially between Cu and Zn or Cu and ∑TCs. Repeated and excessive application of manures from intensive farming systems may produce combined contamination with HMs and TCs which were found in the top 20 cm of the arable soil profiles and also extended to 20–40 cm depth. Increasing manure application rate and cultivation time led to continuing increases in residue concentrations and movement down the soil profile.     

堆肥对污泥中四环素类抗生素及抗性基因的影响

近年来,抗生素及抗性基因的污染与扩散成为全球关注的热点。污泥,作为污水处理系统的副产物,是环境中抗生素抗性基因的重要存储库。污泥经堆肥后可作为肥料施用于农田,然而关于污泥堆肥过程中抗性基因丰度变化的研究却很少。对污泥堆肥过程中四环素类抗生素(四环素、土霉素和金霉素)和四环素类抗性基因(tet(A), tet(C), tet(M), tet(O)和tet(X))的动态变化进行了定量研究。结果表明,四环素、土霉素和金霉素的浓度经堆肥处理后分别减少85.6%, 91.4% 和 85.3%。高温是导致四环素类抗生素降解的主要因素。经堆肥处理后,tet(A)和tet(X)的相对丰度增加,tet(C)、tet(M)和tet(O)的相对丰度略微下降,说明堆肥处理对污泥中四环素类抗性基因的削减效果有限。     

Dissipation of oxytetracycline, chlortetracycline, tetracycline and doxycycline using HPLC-UV and LC/MS/MS under aquatic semi-field microcosm conditions

A mixture of four tetracyclines; oxytetracycline (OTC), chlortetracycline (CTC), tetracycline (TC), and doxycycline (DC) was applied in fifteen 12000l outdoor microcosms at four treatment levels plus controls each with three replicates (n = 3). The dissipation times of parent compounds were monitored and half-lives (DT50) of 1-4 days, depending on treatment level were recorded. This is in accordance with half-lives from previous findings in bench-top experiments. Parent compound DT50, were determined using HPLC-UV. Furthermore, the samples were analyzed for ten different tetracycline products using LC/MS/MS. Two products were found for chlortetracycline; 4-epi-anh-chlortetracyline and the iso-chlortetracycline. Iso-forms were only found for CTC and only at the highest treatment (300 microg l(-1)). The half-lives, trajectories, and relative amounts of the products were analogous for all four tetracyclines. DT50 for products were less than 1.2 days. Formation of 4-epi-anh-tetracyclines, occurred at neutral to weak alkaline conditions.     

Anaerobic digestion of livestock and poultry manures spiked with tetracycline antibiotics

Abstract

We investigated the anaerobic degradation of tetracycline antibiotics (tetracycline [TC], oxytetracycline [OTC] and chlortetracycline [CTC]) in swine, cattle, and poultry manures. The manures were anaerobically digested inside polyvinyl chloride batch reactors for 64?days at room temperature. The degradation rate constants and half-lives of the parent tetracyclines were determined following first-order kinetics. For CTC the fastest degradation rate was observed in swine manure (k?=?0.016?±?0.001 d^?1; half-life = 42.8?days), while the slowest degradation rate was observed in poultry litter (k?=?0.0043?±?0.001 d^?1; half-life = 161?days). The half-lives of OTC ranged between 88.9 (cattle manure) and 99.0?days (poultry litter), while TC persisted the longest of the tetracycline antibiotics studied with half-lives ranging from 92.4?days (cattle manure) to 330?days (swine manure). In general, the tetracyclines were found to degrade faster in cattle manure, which had the lowest concentrations of organic matter and metals as compared to swine and poultry manures. Our results demonstrate that tetracycline antibiotics persist in the animal manure after anaerobic digestion, which can potentially lead to emergence and persistence of antibiotic resistant bacteria in the environment when anaerobic digestion byproducts are land applied for crop production.     

Residues of pharmaceutical products in recycled organic manure produced from sewage sludge and solid waste from livestock and relationship to their fermentation level

In recent years, sludge generated in sewage treatment plants (STPs) and solid waste from livestock being utilized is useful for circulation of nourishment in farmlands as recycled organic manure (ROM). In this study, we determined the residue levels and patterns of 12 pharmaceutical products generated by human activity in the ROMs produced from human waste sludge (HWS), sewage sludge (SS), cattle manure (CM), poultry manure (PM), swine manure (SM) and horse manure (HM).The kind and number of pharmaceutical products detected in ROMs were different. Fluoroquinolones (FQs) were detected at high levels in HWS and SS samples. In addition, the detection frequency and concentration levels of sulfonamides (SAs) in PM and SM were high. Moreover, high concentrations of chlortetracycline (CTC) were found in only SM. These differences reflect specific adherence adsorption of the pharmaceutical products to different livestock and humans. Moreover, it was found that the concentrations of pharmaceutical products and fermentation levels of ROMs had significant positive correlation (r = 0.41, p = 0.024).When the fermentation test of ROM was conducted in a rotary fermentor in a lab scale test, the residue levels of pharmaceutical products decreased effectively except carbamazepine (CBZ). The rates of decrease were in the case of tetracyclines (TCs): 85-92%, FQs: 81-100%, erythromycine: 67%, SAs: 79-95%, trimethoprim: 86% and CBZ: 37% by 30 d. Pharmaceutical products that can be decomposed by fermentation process at the lowest impact of residual antibiotic activities may therefore be considered as environmentally friendly medicines.     

The influence of light exposure,water quality and vegetation on the removal of sulfonamides and tetracyclines: A laboratory-scale study

The effect of aquatic vegetation (Spyrogira sp. and Zannichellia palustris), light exposure and water quality (secondary-treated wastewater vs. ultrapure water) on the removal efficiency of six antibiotics (sulfonamides and tetracyclines) is studied in laboratory-scale reactors. After 20 d of treatment, 3–59% of sulfonamides were eliminated in the reactors exposed to light. Removal was about 10% in unplanted reactors in darkness. The elimination of tetracycline (TC) and oxytetracycline (OTC) ranged between 83% and 97% in both planted and unplanted reactors. However, in dark unplanted reactors, OTC was largely removed (88%) while only 15% of TC was eliminated. These results suggest that TC was mainly removed by photodegradation whereas biodegradation or hydrolysis process seems to be significant processes for OTC. Sulfonamides were mainly eliminated by biodegradation or indirect photodegradation processes. Pseudo-first order kinetics removal rates ranged from 0.003 and 0.007 d^?1 for Sulfamethazine and TC in the covered control reactors to 0.13 and 0.21 d^?1 for TC and OTC in the uncovered control reactors, with half-lives from 3 to 350 d. A TC photodegradation product was tentatively identified in uncovered reactors. This study highlights the important role played by light exposure in the elimination of antibiotics in polishing ponds.     

Characteristics of oxytetracycline sorption and potential bioavailability in soils with various physical-chemical properties

Veterinary antibiotics are widely used for disease treatment, prevention and animal growth promoting. Frequent detection of veterinary antibiotics in environments, caused by land application of untreated or even treated antibiotics-containing animal wastes, has posed the growing concern of their adverse effect on natural ecosystems. Oxytetracycline (OTC) is one of the most widely-used veterinary antibiotics in livestock industry. OTC present as a cation, zwitterions, or net negatively charged ion in soils complicates predicting its sorption characteristics and potential bioavailability and toxicity. This study was to identify soil properties influencing OTC sorption and its subsequent bioavailability in five soils with various physical-chemical properties. A solution used to determine bioavailable analytes in soils and sediments, 1 M MgCl₂ (pH 8.5), was chosen to desorb the potentially bioavailable fraction of OTC sorbed onto soils. Our results demonstrated that soils with higher illite content and permanent cation exchange capacity have higher OTC sorption capacity, but increase the availability of sorbed OTC indicated by higher release of sorbed OTC from soils into aqueous phase in 1 M MgCl₂ (pH 8.5). Reversely, soil organic matter (SOM), clay, kaolinite, variable cation exchange capacity, DCB-Fe and -Al have lower OTC sorption capacity, but decrease the release of sorbed OTC from soils into 1 M MgCl₂. These findings indicate that SOM and clay greatly influence OTC adsorption and potential availability. This study contributes significantly to our understanding of the potential bioavailability of sorbed OTC and the effects of soil properties on OTC sorption behaviors in soils.     

Binding of the veterinary drug tetracycline to bovine hemoglobin and toxicological implications

Tetracycline (TC) is a widely used veterinary drug in animal breeding and fishery. Because of its low bioavailability, the TC residue extensively exists in the environment (e.g. soils, lakes and rivers), which can enter the human body, being potentially harmful. Hemoglobin (Hb) is a protein responsible for oxygen carrying in the vascular system of animals. The aim of this study was to investigate the interaction of bovine hemoglobin (BHb) with TC through spectroscopic and molecular modeling methods. The experimental results revealed that TC can interact with BHb with one binding site to form a TC-BHb complex, mainly through van der Waals interactions and hydrogen bonds. The UV-visible absorption, synchronous fluorescence, and circular dichroism (CD) results revealed that the binding of TC can cause conformational and some microenvironmental changes of BHb, which may affect BHb physiological functions. The synchronous fluorescence experiment disclosed that TC binds into BHb central cavity, which was verified by molecular modeling study. The work contributes to clarify the molecular mechanism of TC toxicity in vivo.     

Methyl-triclosan binding to human serum albumin: Multi-spectroscopic study and visualized molecular simulation

Methyl-triclosan (MTCS), a transformation product and metabolite of triclosan, has been widely spread in environment through the daily use of triclosan which is a commonly used anti-bacterial and anti-fungal substance in consumer products. Once entering human body, MTCS could affect the conformation of human serum albumin (HSA) by forming MTCS–HSA complex and alter function of protein and endocrine in human body. To evaluate the potential toxicity of MTCS, the binding mechanism of HSA with MTCS was investigated by UV–vis absorption, circular dichroism and Fourier transform infrared spectroscopy. Binding constants, thermodynamic parameters, the binding forces and the specific binding site were studied in detail. Binding constant at room tempreture (T = 298 K) is 6.32 × 10³ L mol⁻¹; ΔH⁰, ΔS⁰ and ΔG⁰ were 22.48 kJ mol⁻¹, 148.16 J mol⁻¹ K⁻¹ and −21.68 kJ mol⁻¹, respectively. The results showed that the interactions between MTCS and HSA are mainly hydrophobic forces. The effects of MTCS on HSA conformation were also discussed. The binding distance (r = 1.2 nm) for MTCS–HSA system was calculated by the efficiency of fluorescence resonance energy transfer. The visualized binding details were also exhibited by molecular modeling method and the results could agree well with that from the experimental study.     

纳米TiO2暴露对水中四环素类抗生素光降解行为的影响

本文研究了纳米二氧化钛(TiO₂)暴露对水中土霉素(OTC)、金霉素(CTC)光降解行为的影响。结果表明：纳米TiO₂暴露下OTC、CTC在模拟太阳光下的降解遵循准一级反应动力学模型,光降解效率随纳米TiO₂投加量的增大、粒径的减小和初始pH的升高而逐渐提高。当纳米TiO₂投加量为50.0 mg·L⁻¹、粒径为5 nm,体系初始pH为6.0时,经360 min反应后 OTC、CTC光降解率分别为95.93%、97.13%。溶液中共存的HCO₃⁻对光降解过程起促进作用,而Cl⁻起抑制作用。自由基捕获实验证明·O₂⁻是OTC、CTC光降解过程中起主导作用的活性物种,光降解贡献率分别为91.01%、87.35%。通过毒性实验证明纳米TiO₂长期暴露下可降低OTC、CTC的急性毒性和遗传毒性。     

Evaluation of food chain transfer of the antibiotic oxytetracycline and human risk assessment

There has been recent concern regarding the possibility of antibiotics entering the aquatic food chain and impacting human consumers. This work reports experimental results of the bioconcentration of the antibiotic oxytetracycline (OTC) by the Asian watermeal plant (Wolffia globosa Hartog & Plas) and bioaccumulation of OTC in watermeal and water by the seven-striped carp (Probarbus jullieni). They show, for the first time, the extent to which OTC is able to transfer from water to plant to fish and enter the food chain. The mean bioconcentration factor (dry weight basis) with watermeal was 1.28 × 10³ L kg⁻¹. Separate experiments were undertaken to characterize accumulation of OTC by carp from water and watermeal. These showed the latter pathway to be dominant under the conditions employed. The bioconcentration and biomagnification factors for these processes were 1.75 L kg⁻¹ and 2 × 10⁻⁴ kg g⁻¹ respectively. Using an aqueous concentration range of 0.34–3.0 μg L⁻¹, hazard quotients for human consumption of contaminated fish of 1.3 × 10⁻² to 1.15 × 10⁻¹ were derived.     

Influence of anionic, cationic and nonionic surfactants on adsorption and desorption of oxytetracycline by ultrasonically treated and non-treated multiwalled carbon nanotubes

High adsorption capacity of carbon nanotubes (CNTs) may greatly determine the bioavailability and mobility of organic contaminants. The fate of contaminants adsorbed by CNTs may be substantially influenced by surfactants used both in the synthesis and dispersion of CNTs. The aim of this research was to determine the influence of surfactants (nonionic - TX100, cationic - CTAB and anionic - SDBS) on adsorption and desorption of oxytetracycline (OTC) by multiwalled carbon nanotubes (MWCNTs). The surfactants used had a substantial influence on both adsorption and desorption of OTC. The direction of changes depended clearly on the type of surfactant. In case of anionic SDBS, increased adsorption of OTC by MWCNTs was observed. The presence of TX100 and CTAB decreased the adsorption of OTC by MWCNTs significantly. The increase of OTC adsorption after ultrasonic treatment was observed in case of MWCNTs alone and MWCNTs with SDBS and TX100. However, ultrasonic treatment caused OTC adsorption decrease in the presence of CTAB. The change of pH had also an important effect on OTC adsorption in the presence of surfactants. Depending on the surfactant and pH, an increase or decrease of OTC adsorption was observed. The presence of surfactants increased OTC desorption from MWCNTs significantly as follows: SDBS = CTAB < TX100. The results obtained suggest new potential threats and constitute a basis for further research considering the bioavailability and toxicity of antibiotics in the presence of MWCNTs and surfactants.     

Occurrence, distribution and seasonal variation of antibiotics in the Huangpu River, Shanghai, China

Water samples were collected from 19 sampling sites along the Huangpu River in June and December 2009. The occurrence, distribution and seasonal variation of 22 antibiotics, including four tetracyclines, three chloramphenicols, two macrolides, six fluoroquinolones, six sulfonamides and trimethoprim were investigated. It was found that all 19 sampling sites were contaminated by antibiotics. Four antibiotics (sulfamerazine, norfloxacin, fleroxacin and sarafloxacin) were not detected. The detection frequencies of the other 18 antibiotics were in the range of 5.3-100%. The median concentrations of the detected antibiotics ranged from quantification limits to 36.71 ng L⁻¹ (tetracycline) in June and to 313.44 ng L⁻¹ (sulfamethazine) in December. The number of detected antibiotics and the overall antibiotic concentrations were higher in December than in June due to the different river flow conditions. Different dominant antibiotics were observed for each group of antibiotics between June and December. Higher total concentrations of veterinary antibiotics such as tetracyclines were observed in suburban sampling sites than in unban sites, indicating the role of livestock and agricultural activities as an important source of antibiotic contamination.     

Characterization of dissolved organic matter using high-performance liquid chromatography (HPLC)-size exclusion chromatography (SEC) with a multiple wavelength absorbance detector

High-performance liquid chromatography-size exclusion chromatography (HPLC-SEC) coupled with a multiple wavelength absorbance detector (200-445 nm) was used in this study to investigate the apparent molecular weight (AMW) distributions of dissolved organic matter (DOM). Standard DOM, namely humic acid, fulvic acid and hydrophilic acid, from the Suwannee River were tested to ascertain the performance and sensitivity of the method. In addition to four compounds groups: humic substances (Peak 1, AMW 16 kD), fulvic acids (Peak 2, AMW 11 kD), low AMW acids (Peak 3, AMW 5 kD), and low AMW neutral and amphiphilic molecules, proteins and their amino acid building blocks (Peak 4, AMW 3 kD), an new group that appears to include low AMW, 6-10 kD, humic substances was found based on investigating the spectra at various elution times. The spectroscopic parameter S_>365 (slope at wavelengths >365 nm) was determined to be a good predictor of the AMW of the DOM. The detector wavelength played an important role in evaluating the AMW distribution. For some fractions, such as the humic and low AMW non-aromatic substances, the error in measurement was ±30% as determined by two-dimensional chromatograms detected at an artificially selected wavelength. HPLC-SEC with multiple wavelength absorbance detection was found to be a useful technique for DOM characterization. It characterized the AMW distributions of DOM more accurately and provided additional, potentially important information concerning the properties of DOM with varying AMWs.     

Electrochemical oxidation of nitrogen-heterocyclic compounds at boron-doped diamond electrode

Nitrogen-heterocyclic compounds (NHCs) are toxic and bio-refractory contaminants widely spread in environment. This study investigated electrochemical degradation of NHCs at boron-doped diamond (BDD) anode with particular attention to the effect of different number and position of nitrogen atoms in molecular structure. Five classical NHCs with similar structures including indole (ID), quinoline (QL), isoquinoline (IQL), benzotriazole (BT) and benzimidazole (BM) were selected as the target compounds. Results of bulk electrolysis showed that degradation of all NHCs was fit to a pseudo first-order equation. The five compounds were degraded with the following sequence: ID > QL > IQL > BT > BM in terms of their rates of oxidation. Quantum chemical calculation was combined with experimental results to describe the degradation character of NHCs at BDD anode. A linear relationship between degradation rate and delocalization energy was observed, which demonstrated that electronic charge was redistributed through the conjugation system and accumulated at the active sites under the attack of hydroxyl radicals produced at BDD anode. Moreover, atom charge was calculated by semi empirical PM3 method and active sites of NHCs were identified respectively. Analysis of intermediates by GC-MS showed agreement with calculation results.     

Residual veterinary antibiotics in swine manure from concentrated animal feeding operations in Shandong Province, China

The scientific interest in the occurrence and fate of antibiotics in animal husbandry has increased during the past decades because of the emergence and development of antimicrobial resistance in pathogenic bacteria. This study developed a method for simultaneous detection of five sulfonamides, three tetracyclines and one macrolide in swine manure with stable recoveries (73.0-110.6%) and high sensitivity (limit of quantification <90 μg kg⁻¹). Thereafter, a total of 126 swine manure samples, collected from 21 concentrated animal feeding operations (CAFOs) in Shandong Province of China during summer and winter, were analyzed. The potential influences of different sampling seasons, swine types and food sources on residual antibiotic concentrations were examined in detail. The maximum concentration of residual antibiotic could reach up to 764.4 mg kg⁻¹ (chlortetracycline), and the detection frequencies were 84.9-96.8% for tetracyclines, 0.8-51.6% for sulfonamides and 4.8% for macrolide. These data reveal that antibiotics were extensively used in CAFOs in this district and the manure may act as a non-specific source of antibiotic residue in farmlands and aquatic environments.     

Discrepancies in the acute versus chronic toxicity of compounds with a designated narcotic mechanism

In this study, it was illustrated that even for certain simple organic compounds with a designated mode of action (MOA) (i.e. narcotic toxicity) unexpected differences in acute and chronic toxicity can be observed. In a first part of the study, species sensitivity distributions (SSDs) based on either acute or chronic toxicity data of three narcotic test compounds (methanol, ethanol and 2-propanol) were constructed. The results of the acute SSDs were as expected for narcotic compounds: rather similar sensitivity and small differences in toxicity were observed among different species. On the contrary, the chronic SSDs of methanol and ethanol indicated larger interspecies variation in sensitivity. Furthermore, the chronic toxicity trend (ethanol > methanol > 2-propanol) was unexpectedly different from the acute toxicity trend (2-propanol > ethanol > methanol) and acute versus chronic extrapolation could not be successfully described for methanol and ethanol using an ACR of 10 (as suggested for narcotic compounds). In contrast to the interspecies approach in the first part of this study, the second part of the study was focused on the assessment of acute and chronic toxicity of the three test compounds in Daphnia magna, which was identified as one of the most sensitive organisms to methanol and ethanol. Here, the differences in acute and chronic toxicity trend were in accordance to the results of the SSDs. The enhancement of membrane penetration due to the small molecular size of methanol and ethanol, in combination with the higher toxicity of their respective biotransformation products were suggested as potential causes of the increased chronic toxicity. Furthermore, it was stressed that larger awareness of these irregularities in acute to chronic extrapolations of narcotic compounds is required and should receive additional attention in further environmental risk assessment procedure.