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1

Comparison of Sensitivity of Autotrophic and Heterotrophic Microorganisms to the Pollution of Natural Water with Rare Earth Elements (Lanthanum and Cerium)

Lozhkina Roza A., Olkova Anna S., Koval Ekaterina V., Tomilina Irina I., Sysolyatina Maria A.

Journal of Ecological Engineering

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2022

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Vol. 23, nr 4

58--63

EN

The purpose of the work was to compare the sensitivity of autotrophic and heterotrophic organisms used in bioassay to lanthanum and cerium under the conditions of a model experiment with aqueous media. Using bioassay methods, the pre-lethal effects of La and Ce in heterotrophic Paramecium caudatum and Escherichia coli, as well as autotrophic Chlorella vulgaris and Nostoc linckia were determined. Model solutions of La2 (SO4)3∙8H2O and Ce2 (SO4)3∙8H2O were tested in the concentration range of 0.1–200 mg/l. As a result, it was shown that heterotrophic organisms are more sensitive to water pollution with La and Ce than autotrophic ones. According to the totality of experiments, cerium turned out to be more toxic than lanthanum. When planning the environmental studies of wastewater or reservoirs polluted with REE, it is recommended to focus on comparative sensitivity of bioassay methods, taking into account the test-functions used: bioassay for chemotaxis of P. caudatum > bioassay for changes in bioluminescence of E. coli (strain M-17) > bioassay for the content of chlorophyll а and intensity of lipid peroxidation in N. linckia > bioassay on the increase in the number of Cl. vulgaris.

2

Bioremediation Possibilities of Oil-Contaminated Soil by Biosurfactant Based on Bacillus Strain

Nedoroda Vladyslav, Trokhymenko Ganna, Magas Nataliya

Journal of Ecological Engineering

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2022

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Vol. 23, nr 8

49--55

EN

This paper represents the results of the investigation of soil phytotoxicity with a high level of oil pollution. The artificially contaminated soils were exposed to an oil-destroying biosurfactant based on the strains of Bacillus amyloliquefaciens and Bacillus subtilis. The main objective of the work was to evaluate the effectiveness of a consortium of microorganisms in reducing the phytotoxicity of soils with high levels of oil contamination. The determination was performed on the reactions of test organisms to the pollution of the soil environment. The test system in the experiment comprised Sorghum bicolor subsp. Drummondii, Phleum pretense, Galéga officinális, Trifolium pretense, Medicágo sativa, and Sinapis arvensis. The variety of the applied indicator plants has provided a comprehensive analysis of the results of oil destruction and allowed an assessment of the sensitivity of the seeds of each species to toxic substances.

3

Behavioral and Lethal Effects of La Salt and a Mixture of Cu and La Salt on Daphnia magna Straus

Olkova Anna Sergeewna, Sysolyatina Maria

Journal of Ecological Engineering

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2022

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Vol. 23, nr 6

245--252

EN

The study of joint effects of REE and HM is relevant, since they are often satellite deposits, their areas of application are similar, and an increase in concentrations of elements of these groups in the areas that are not places of their extraction and enrichment is proven. The purpose of this work was to find out the pre-lethal and lethal effects of La, Cu and their equimolar mixtures in tests for Daphnia magna Straus. Bioassays of artificially polluted natural waters, initially free of toxic elements, was carried out. In bioassays on the mortality of D. magna in the space of 96 hours it was found that acute toxicity of copper sulfate solutions is observed at the calculated concentration of Cu2+ 0.1 mg/L (0.0016 mmol/L), and the acute toxicity of lanthanum sulfate is when the dose of La3+ is equal to 50 mg/L (0.36 mmol/L). In the solutions comprising mixtures of Cu and La salts (1:1 calculated using metals), the concentrations of which are equimolar to the investigated solutions of copper sulfate, the mortality of D. magna begins in the solution containing 10 times less toxic elements. It was found that 25% of individuals died in the variant “0.00016 mmol/L”, the mortality of 100% of individuals was at the total metal concentration of 0.0008 mmol/L. The solutions containing La (0.072–0.72 mmol/L) and Cu (0.00016–0.0016 mmol/L) naturally inhibit the motor activity of D. magna by 1.3–5.3 times and 1.2–1.9 times in 1 hour and 1.7–2.8 and 1.4–2.2 times in 24 hours, respectively. The solutions containing mixtures of Cu and La salts inhibited the motor activity of D. magna in the same way as copper sulfate solutions with the Cu2+ concentrations equimolar “Cu2+ + La3+”. Therefore, when testing the solutions with the same molar concentrations of Cu2+ and the mixture of “Cu2+ + La3+” it was shown that La potentiates the pre-lethal effect of Cu to the level of individual effects of Cu. The additions of La salt to the solutions containing pre-lethal doses of Cu lead to lethal effects of such mixtures for D. magna.

4

Targeted Selecting of Bioassay Methods as an Alternative to the "Battery of Bioassays"

Olkova Anna

Ecological Engineering & Environmental Technology

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2022

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Vol. 23, iss. 2

153--161

EN

The article describes an algorithm for selecting a targeted bioassay method. It was proposed to consider the most sensitive method to a certain contamination as the targeted method of bioassay. The selection was made from a "battery of bioassays", which necessarily includes the D. magna mortality test. The rest of the methods for comparing sensitivity were selected situationally. The algorithm was tested on model samples. It was found that mortality tests for D. magna and C. affinis are the most sensitive to contamination with mineral nitrogen compounds. E. coli bioluminescence reduction test is preferred when there is contamination with Cu, phosphates and pyrophosphates. It was shown that the test to reduce the chemotactic reaction of P. caudatum should be used when the aquatic environment is polluted with Cd, Pb, Zn, oil products, organic herbicides imazethapyr and imazamox. The proposed algorithm is universal, but it should be applied when the priority pollutant is known, the effects of which prevail over the action of other compounds in the sample.

5

Liquid chromatographic enantioseparation, determination, bioassay and isolation of enantiomers of Ketorolac : a review

Bhushan Ravi

Acta Chromatographica

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2022

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Vol. 34, no. 3

220--236

EN

Ketorolac (Ket) is a potent non-narcotic analgesic drug (among the nonsteroidal anti-inflammatory drugs). The physiological activity of Ket resides with (S)-(-)-Ket while the drug is marketed and administered as a racemic mixture. Therefore, it is desirable that the pharmacokinetics is measured and quantified for enantiomers individually and not as a total drug. The present paper is focused on relevant literature on LC enantioseparation of (RS)-Ket along with bioassay, pharmacokinetic and clinical studiem within the discipline of analytical chemistry. HPLC and Thin layer chromatography (TLC) methods using both direct and indirect approaches are discussed. The methods provide chirality recognition even in the absence of pure enantiomers. Besides, a brief discussion on resolution by crystallization and enzymatic methods is included. The most interesting aspects include establishment of structure and molecular asymmetry of diastereomeric derivatives using LC-MS, proton nuclear magnetic resonance spectrometry, and by drawing conformations in three dimensional views by using certain software. A brief discussion has also been provided on the recovery of native enantiomers by TLC.

6

The Use of the Triad Approach to Ecological Assessment of the State of a Reservoir Receiving Wastewater From a Thermal Power Plant

Mahanova Elena

Journal of Ecological Engineering

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2021

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Vol. 22, nr 2

155--160

EN

Thermal power plants (TPP) belong to the enterprises that accompany the development of any city. These industrial facilities consume and discharge large amounts of water. Therefore, thermal power plants are often located near water bodies that need to assess their ecological state and predict the consequences of the anthropogenic impact. The purpose of the work was to assess the ecological state of the floodplain Lake Ivanovskoye (Russia, Kirov region), which receives the wastewater from a thermal power plant, using a triadic approach and comparing the results of chemical analysis, bioassay and bioindication. It was shown that the main pollutant in the lake and the canal connecting Lake Ivanovskoye with the nearest river Vyatka is ammonium ion (up to 3.2 mg/dm3). In addition, the Russian standards for biological oxygen consumption are exceeded (analysis period is 20 days). The tests for the toxicity of wastewater showed inhibition of Paramecium caudatum and Escherichia coli reactions. In contrast, natural surface water from Lake Ivanovskoye stimulated the test functions of Scenedesmus quadricauda, P. caudatum, and E. coli. The death of Daphnia magna in the samples were not recorded. The totality of the facts pointed to the organic nature of pollution, leading to the anthropogenic eutrophication of the lake. The hypothesis was tested by using the bioindication method and by analyzing the totality of species of coastal aquatic plants. It was shown that the reservoir belongs to the mesotrophic type with a moderate degree of pollution, which decreases with distance from the wastewater discharge points. The increase in the trophicity of the reservoir is facilitated by thermal pollution (up to +15℃) and an alkaline pH of wastewater (up to 9.2). Thus, the joint analysis of the results of chemical analysis, bioassay and bioindication made it possible to carry out an objective assessment of the ecological state of the reservoir and identify the reasons for the revealed facts.

7

Intraspecific Sensitivity to Toxicants – a Methodological Problem of Bioassay: Review

Olkova Anna

Journal of Ecological Engineering

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2021

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Vol. 22, nr 7

113--122

EN

The article discusses the scientific studies that reveal the mechanisms of intraspecific differences in responses of organisms to chemical exposure. The factors of individual sensitivity to toxicants are represented by genetic differences between individuals of the same species and genetically unconditioned factors. The genetic factors are mutations, including under-researched mitochondrial DNA mutations and genomic drift, genetic polymorphism, and gender-related differences. Age, conditions of keeping and state of organisms, seasonal variations of body functions, and biotic interactions are considered as the factors that are not related to the genetic apparatus. The article considers a number of works, in which the effects of the combined action of external factors on sensitivity of organisms in model groups are studied.

8

"Battery of Bioassays" for Diagnostics of Toxicity of Natural Water when Pollution with Aluminum Compounds

Olkova Anna, Berezin Grigory

Journal of Ecological Engineering

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2021

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Vol. 22, nr 2

195--199

EN

The article presents the results of an experimental comparison of the sensitivity of biotests using Daphnia magna Straus, Ceriodaphnia affinis Lilljeborg, Paramecium caudatum Ehrenberg, and Escherichia coli Migula (strain M-17) to water pollution with aluminum compounds. The research was carried out under simulated conditions: the model toxicant was aluminum sulfate Al2(SO4)3∙18H2O, the concentration range per Al was 0.04–2.8 mg/dm3, and the pH of the tested waters was close to the neutral level of 7.2–7.8. The bioluminescence of E. coli significantly decreased at an Al concentration of 0.8 mg/dm3 (toxicity index was 93.3±1.2, which refers to a high level of toxicity). The reaction of P. caudatum was weaker: a high level of toxicity was achieved at an Al concentration of 2.8 mg/dm3. These doses did not cause the death of D. magna and C. affinis in short-term experiments (28 and 96 hours, respectively). However, in the tests for the chronic toxicity of aluminum, we showed that the doses of 0.8 and 2 mg/dm3 Al cause high death of individuals (more than 50%) and a significant decrease in the number of offspring. The range of sensitivity of the bioassay methods to water pollution with aluminum turned out to be as follows: bioassay for the bioluminescence of E. coli > bioassay for the changes in chemotaxis of P. caudatum > bioassay for the changes in fertility of D. magna > bioassay for the changes in fertility of C affinis.

9

Transformation of Urbanozems in the Areas of Gas Stations

Olkova Anna, Zimonina Natalia

Ecological Engineering & Environmental Technology

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2021

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Vol. 22, iss. 3

51--58

EN

The areas near gas stations are places of local specific environmental pollution. The purpose of this work was to study the environmental consequences of gas station functioning on the surrounding soils (urbanozems). The content of oil products (OP), bulk and mobile forms of heavy metals, acute toxicity (Ceriodaphnia affinis Lillieborg, Раramecium caudatum Ehrenberg, Escherichia coli Migula), chronic toxicity (C. affinis), and the state of soil microphotrophs were determined. The content of OP was from 520±130 to 4820±100 mg/kg, which is significantly higher than the concentrations of OP in the soils of the transport zone of cities. The exceeding of the Russian standards for HM was found only for zinc (the maximum for the total form – 1.7 MPC, for the mobile form – 1.4 MPC). The urbanozems samples did not have acute toxicity; however, chronic toxicity in the bioassay for the mortality of C. affinis (up to 85%) and a decrease in the fertility of crustaceans (2–3 times compared to the control) were observed. In the structure of the community of soil microphototrophs, cyanobacteria dominated both in biomass and in abundance: (up to 748±10 thousand cells/1 g of soil) compared with green algae and diatoms (1.1–5.5 and 1.2–19.8 thousand cells/1 g of soil, respectively). As a result of the work, it was concluded that the quantitative algological analysis and biotest with the assessment of chronic toxicity in terms of mortality and fertility of C affinis for the diagnosis of local soil contamination in the areas of gas stations were of the highest informative value.

10

Toxicity of Water after Short-Term Contact with PVC Materials Depending on the Temperature and Components of the Polymer Composition

Olkova Anna

Ecological Engineering & Environmental Technology

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2021

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Vol. 22, iss. 5

119--125

EN

This work shows a paradoxical fact: a highly plasticized PVC sample containing the maximum amount of the plasticizer di(2-ethylhexyl) phthalate (DEHP) compared to medium- and low-plasticized polymers turned out to be the least toxic. This was confirmed in express bioassays for the pre-lethal reactions of Paramecium caudatum Ehrenber and Escherichia coli M-17 when determining the acute toxicity for mortality of Daphnia magna Straus and Ceriodaphnia affinis Lilljeborg, as well as in chronic tests for D. magna. The migration of DEHP from the tested PVC samples into water was confirmed by the mass spectrometry method. It was shown that the contact of PVC materials with hot water (70℃) led to a significant increase in the toxicity of the extracts compared to cold extraction (20℃). The search for safe composition of PVC materials must be carried out in the direction of the optimal ratio of PVC-polymer, plasticizer and heat stabilizer, as well as replacing the components of the composition with safe substances.

11

Assessment of the Toxicity of the Natural and Technogenic Environment for Motor Activity of Daphnia magna

Olkova Anna, Zimonina Natalia

Journal of Ecological Engineering

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2020

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Vol. 21, nr 7

11--16

EN

The Daphnia magna Straus (1820) crustaceans are used in many countries to assess the quality of the environment. Researchers are guided more often by the mortality of individuals. However, the sublethal effects contribute to the development of biological early warning systems (BEWS). A visual method for recording the motor activity of D. magna was proposed. This method has advantages over automated analogs. It is simple, accessible to performers and does not require the use of special instruments. The method was tested under conditions of modeling the lethal and sublethal effects of heavy metal salts (Cu and Zn). The diagnosis of lethal doses is possible after 1 hour of exposure, whereas the death of crustaceans can occur in 3–4 days of the experiment. The effect of sublethal doses of heavy metals becomes statistically significant after 24 (p < 0.05). The sensitivity of the method was confirmed in the studies of the aqueous extracts from the soils contaminated with various heavy metals (heavy metal processing area). The method showed good results in assessing the toxic effect of the waters anthropogenically contaminated with the mineral forms of nitrogen (NH4+and NO3-) in the region of fertilizer production. The maximum inverse correlation between the motor activity and the concentration of ammonium ions in water (-0.83) was shown after 24 hours. Thus, the method of visual assessment of motor activity can be used in environmental monitoring.

12

A. niger susceptibility of HDPE/hemp composites. Part 1: Resistance to growth of A. niger on the surface

Cofta G., Doczekalska B., Bujnowicz K.

Annals of Warsaw University of Life Sciences - SGGW. Forestry and Wood Technology

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2018

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No. 104

109--112

EN

In the present study the composites of polyethylene and hemp were prepared. In addition the polyethylene-hemp composites containing a Polybond 3039 were also obtained. The objective of this study was to evaluate the resistance of PE-hemp composites to attack by mold. At weekly intervals, composites was evaluated visually intensity of the growth of A. niger using the scale on a surface in accordance with ISO-846. It was found that A. niger grew the surface of composites within two weeks. The use of the Polybond 3039 (coupling agent) reduces the resistance of composites to A. niger. Fungus grew faster on the surface of the PE70 H30 P5 composite samples than the composite designated as PE65 H30.

13

Zastosowanie biotestów we wstępnej ocenie jakości wody basenu szkolnego

Łaskawiec E., Kudlek M., Dudziak M., Wyczarska-Kokot J.

Gaz, Woda i Technika Sanitarna

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2016

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Nr 9

330--334

PL

W pracy przedstawiono oddziaływanie związków chemicznych, w tym ubocznych produktów dezynfekcji (DBP) występujących w wodach basenowych na wybrane organizmy wskaźnikowe. Ocenę toksykologiczną poprzedzono krótką analizą podstawowych parametrów fizykochemicznych. Wykonane biotesty obejmowały zróżnicowaną grupę organizmów, tj. bakterie Aliivibrio fischeri, skorupiaki Daphnia magna, larwy owadów Chaoborus fahicans oraz rośliny naczyniowe Lemna minor. Analizowano korelację pomiędzy wynikami testów, co w przyszłości umożliwi wybór organizmów wskaźnikowych przydatnych w ocenie jakości wód basenowych.

EN

The study presents the impact of chemical substances present in swimming pool water, including disinfection by-products (DBP), on selected indicator organisms. Toxicological assessment was preceded by brief analysis of basic physicochemical parameters. The performed biotests involved a varied group of organisms, including bacteria (Aliivibrio fischeri), crustaceans (Daphnia magna), insect larvae (Chaoborus falvicans), and vascular plants (Lemna minor). Consideration was given to correlations between the results of the tests that will facilitate future selection of indicator organisms to be used in the assessment of quality of swimming pool waters.

14

Biotesty - narzędzia do oceny stopnia zanieczyszczenia środowiska

Wieczerzak M., Namieśnik J.

Analityka : nauka i praktyka

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2014

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nr 3

52--55

15

Współczesna bioanalityka użytecznym narzędziem w ocenie jakości powietrza

Cieszyńska M., Wilma K., Gałęzowska G., Wolska L.

Analityka : nauka i praktyka

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2013

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nr 2

28--37

PL

Wykorzystanie biotestów stanowi ogromny potencjał badawczy, który wnosi dużo dodatkowych informacji uzupełniających tradycyjną analitykę chemiczną i dlatego powinnien być rozwijany i udoskonalany.

16

Toksyczność chlorku 1-heksylo-3-metyloimidazoliowego względem wybranych organizmów wodnych

Arendarczyk A., Zgórska A., Grabińska-Sota E.

Inżynieria i Ochrona Środowiska

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2011

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T. 14, nr 2

137-143

PL

Dokonano oceny potencjału toksykologicznego imidazoliowej cieczy jonowej o nazwie chlorek 1-heksylo-3-metyloimidazoliowy (CHM). Zakres badań ekotoksykologicznych obejmował przeprowadzenie testu toksyczności ostrej z wykorzystaniem skorupiaków słodkowodnych (Thamnocephalus platyurus) i testu toksyczności chronicznej z wykorzystaniem bakterii morskich (Vibrio fischeri). Wartości wskaźników ekotoksykologicznych wyznaczone w poszczególnych testach wyniosły odpowiednio: EC50 2,51 mg/dm3 (THAMNOTOXKIT FTM) i EC50/5min 15,30 mg/dm3 i EC50/15min 14,40 mg/dm3 (MICROTOX® Basic Test 81,9%). Na podstawie uzyskanych wyników związek sklasyfikowano jako toksyczny. W ramach badań dokonano również oceny wpływu korekty odczynu i napowietrzania na redukcję toksyczności roztworu CHM. Doświadczenie przeprowadzono zgodnie z metodyką US EPA/600/6-91/005F. Stwierdzono, że korekta odczynu roztworu CHM do poziomu 3 i 11, a także przeprowadzenie symultanicznego procesu korekty pH do 3 i napowietrzania wpływa na obniżenie toksyczności badanego związku.

EN

Ionic liquids (IL's) are defined as organic salts with low melting point. The reputation of those substances as green solvents (environmental friendly chemicals) is based primarily on their negligible vapor pressure. However, high solubility and stability of those substances in water can cause a serious concern about their toxic potential. Relatively little is still known about the toxicity of these substances as a class, especially when compared to conventional organic solvents. The aim of the study was to investigate the toxicity of imidazolium ionic liquid: 1-heksyl-3-methylimidazolium chloride (CHM) according to selected aquatic organisms: bioluminescent marine bacterium and crustacean. The results showed that CHM caused a harmful effect on tested organisms in the whole range of concentrations used in the tests. The estimated toxic index EC50 obtained 2.51 mg/dm3 according to Thamnocephalus platyurus and EC50/5min 15.30 mg/dm3, EC50/15min 14.40 mg/dm3 according to Vibrio fischeri. The CHM was classified as a toxic substance. Moreover, the toxicity of CHM samples with modified pH values and aeration has been measured. Modifications of CHM samples were based on U.S. EPA Protocol (Toxicity Identification Evaluation). Acute toxicity of each modification have been determined by using the MICROTOX® Basic Test 81.9% standard procedure. The results showed that upgrading pH value to 11 as well as diminishing samples pH to value 3 had an influence on samples' post-reaction toxicity. Both processes, pH correction and aeration, used individually caused a significant decrease of analyzed samples' toxicity expressed as toxic index's EC50 measured after 5 and 15 minutes of exposition. The toxic index values were increased from range 30% (pH 3) up to 100% (pH 11) according to non modified sample. The study showed that pH correction to value 3 connected with simultaneous aeration additionally decrease sample toxicity. The results of the present study has proved that 1-heksyl-3methylimidazolium chloride could be toxic to aquatic organisms and it is necessary to carry out the ecotoxicological risk of ionic liquids by the use of bioassays.

17

Biotest as an indicator of dioxin-like PCBs presence

Małagocki P., Piskorska-Pliszczyńska J.

Proceedings of ECOpole

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2010

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Vol. 4, No. 2

267--272

EN

Dioxins (polychlorinated dibenzo-p-dioxins, PCDDs, polychlorinated dibenzofurans, PCDFs) and dioxin-like polychlorinated biphenyls (dl-PCBs) are toxic compounds which are commonly present in the food chain. According to the European Union regulations the presence of these compounds in food may be determined by complementary and screening and methods considered confirmatory. The combination of the two methods allows a relatively fast selection of food and feed samples containing dioxins at concentrations exceeding acceptable levels and permits an unequivocal confirmation of tested compounds. The aim of the study was to evaluate the usefulness of the biotest based on genetically modified cell lines sensitive to PCDDs and PCDFs for the simultaneous detection and quantitative determination of dl-PCBs in food samples. Mouse hepatoma cell lines (Hepa1L6.1c3) with the luciferase reporter gene were used. The principle of the biotest involving the dioxin mechanism of action includes the presence of receptor Ah agonist Hepa1L6.1c3 cell line synthesize luciferase at concentrations proportional to the agonist dose. The measurement of luciferase activity for various 2,3,7,8-TCDD concentrations (calibration curve) permits quantitative measurements of the Ah receptor antagonist content in examined food samples. The extraction of dl-PCBs from matrixes (salmon, herring and sprat muscles) was performed using the extraction and purification procedure for PCDDs/PCDFs analysis. Separation of dl-PCB from PCDD/PCDF was done by a column chromatography. The concentration of dl-PCBs was determined using the biotest. The results involving the same samples were compared with those obtained by the HRGC/HRMS method regarded as confirmatory. The results confirm the biotest as a useful method for dl-PCBs determinations; the results obtained by the two methods are parallel and meets the criteria defined by the Commission Regulation (EC) No. 1881/2006.

PL

Dioksyny (polichlorowane dibenzo-p-doksyny (PCDD) i polichlorowane dibenzofurany (PCDF)) oraz dioksynopodobne polichlorowane bifenyle (dl-PCB)) są związkami toksycznymi występującymi m.in. w łańcuchu żywieniowym. Zgodnie z przepisami Unii Europejskiej, do oznaczania ich zawartości w żywności można zastosować metody komplementarne: przesiewową i potwierdzającą. Ich połączenie pozwala na stosunkowo szybką selekcję próbek żywności i pasz zawierających dioksyny w stężeniach przekraczających dopuszczalne poziomy oraz pozwala na jednoznaczne potwierdzenie obecności badanych związków. Celem pracy było określenie przydatności biotestu bazującego na genetycznie zmodyfikowanej linii komórek wrażliwych na działanie PCDD i PCDF do równoczesnego wykrywania i ilościowego oznaczania dl-PCB w próbkach żywności. W badaniach zastosowano linię komórkową hepatomy mysiej (Hepa1L6.1c3) z wprowadzonym transgenem lucyferazy pod kontrolą receptora Ah. Zasada biotestu wykorzystującego mechanizm działania dioksyn jest następująca: w obecności agonistów receptora Ah komórki Hepa1L6.1c3 syntezują enzym lucyferazę w stężeniu proporcjonalnym do dawki agonisty. Pomiar aktywności lucyferazy wobec serii stężeń 2,3,7,8-TCDD (krzywa kalibracyjna) pozwala na ilościową ocenę zawartości agonistów receptora Ah w badanym ekstrakcie próbki żywności. Do ekstrakcji dl-PCB z matrycy (mięśnie łososia, śledzia, szprota) użyto zoptymalizowanej dla PCDD/PCDF metody ekstrakcji i oczyszczania. Na kolumnach chromatograficznych oddzielano dl-PCB od PCDD/PCDF. Zawartość dl-PCB oznaczano, stosując biotest. Wyniki uzyskane biotestem porównywano z rezultatami oznaczeń chemiczną metodą potwierdzającą HRGC/HRMS, wykonaną w tych samych próbkach. Uzyskane dane pozwalają ocenić biotest jako przydatne narzędzie w badaniach zawartości dl-PCB, ponieważ wyniki uzyskane obydwoma metodami są porównywalne, a metoda spełnia kryteria określone przepisami prawa wspólnotowego (Rozp. 1881/2006/WE).

18

The first Polish interlaboratory comparison of the luminescent bacteria bioassay with three standard toxicants

Nałęcz-Jawecki G., Baran S., Mankiewicz-Boczek J., Niemirycz E., Wolska L., Knapik J., Piekarska K., Bartosiewicz M., Piętowski G.

Environment Protection Engineering

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2010

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Vol. 36, nr 3

95-102

EN

Quality assurance test with standard toxicants should be carried out regularly to check the sensitivity of the testbiont and the quality of the procedure. In the luminescent bacteria bioassay, zinc sulphate and phenol have been used as the respective inorganic and organic reference sub- stances. ISO 11348 standard proposes 3,5-dichlorophenol (DCP) as a standard toxicant for luminescent bacteria toxicity assay. This work presents the results of the first ring study performed by 9 Polish laboratories. One hundred and twenty five valid toxicity data were received and only 7.2% data were rejected. In the case of DCP, all results were valid and the coefficient of variation for this compound was the lowest.

19

Evaluation of the recombinant cell-based bioassay as a screening method for PCDD/Fs monitoring in fish tissue

Stypula-Trebas S., Warenik M., Gembal M., Malagocki P., Piskorska-Pliszczynska J.

Environmental Biotechnology

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2009

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Vol. 5, No. 1

32-41

EN

Among 210 congeners only 17 highly toxic, 2,3,7,8-chlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) are of toxicological concern. They exhibit high binding affinity to an intracellular aryl hydrocarbon receptor (AhR), causing harmful effects at exposure levels of thousands of times lower than most environmental toxicants. The Chemically Activated LUciferase gene expression bioassay (CALUX) utilizes recombinant cells that were transfected with a luciferase reporter gene, which responds to dioxin-like compounds with the induction of luciferase in a time-, dose-, AhR-dependent and chemical-specific manner. The bioassay evaluation concerning the European Community (EC) requirements for the PCDD/Fs determination for the official control of foodstuffs was performed on salmon tissue. In order to evaluate the bioassay performance characteristics, recovery range, limit of detection (LOD), limit of quantification (LOQ) and precision were determined. The results revealed that combining a dichloromethane: hexane extraction, an acid silica plus activated carbon clean-up provides reliable, reproducible (CV = 9-20%) measurements with acceptable recovery (78%) and sensitivity at the required ppt range. Due to the low cost and high throughput characteristics of the CALUX assay, food monitoring for PCDD/Fs may benefit from use of this bioassay as a prescreening tool to select and prioritize samples for subsequent analysis by high resolution gas chromatography/high-resolution mass spectrometry (HRGC/HRMS). Although the bioassay may not be able to specify identity of the reactive substances, it may serve as a very useful tool for the evaluation of contamination sources.