Endocrine disrupting potency and toxicity of novel sophorolipid quaternary ammonium salts.

A new class of biosurfactants, namely quaternary ammonium sophorolipids (SQAS), suitable for pharmaceutical applications, was tested for the evaluation of their (anti)estrogenic and (anti)androgenic potency with the help of YES/YAS assays. Also their toxicity towards yeasts (Saccharomyces cerevisiae) and bacteria (Escherichia coli) was checked. The results achieved for SQAS, which can be regarded as potential micropollutants, were compared with those obtained for two well-known micropollutants diclofenac and 17α-ethinylestradiol subjected to the same testing procedures. This work demonstrated that acetylation of the hydroxyl group of the carbohydrate head of SQAS decreased the toxicity of this class of biosurfactants towards Saccharomyces cerevisiae. Furthermore, it contributed to the decrease of their endocrine disrupting potency. None of the SQAS studied showed clear agonist activity for female or male hormones. SQAS1 and SQAS2 revealed weak antiestrogenic and antiandrogenic potency. All of these properties were weaker, not only to the potency of the appropriate positive control in the antagonists bioassays, but also compared to the potency of other tested compounds, i.e. DCF and EE2. SQAS3 possessed most probably inhibitory activity towards male hormones. Moreover, cytotoxicity of two out of four studied SQAS at the highest concentrations towards the strains of Saccharomyces cerevisiae interfered with the endocrine disruption activity. It would be also worth studying it with the use of another endocrine activity test.

Adaptation of microbial communities in activated sludge to 1-decyl-3-methylimidazolium bromide.

The effects of 1-decyl-3-methylimidazolium bromide on activated sludge process and microbial composition were investigated. Ionic liquid (IL) was dosed continuously to the laboratory activated sludge system at an influent concentration from 1 to 20 mg l(-1) for about 1 month. As compared to the control test, mean values of degree of chemical oxygen demand removal and degree of biochemical oxygen demand removal were almost remaining constant at a high level, equaling 92.6% and 98.1%, respectively. In addition, no influence of IL on size and shape of flocs was observed. The values of the sludge biotic index indicate that sludge exposed on IL was stable and very well colonized with good biological activity. Increases in Proteobacteria (mainly Variovorax sp., Vogesella sp., Hydrogenophaga sp.), Bacteroidetes (mainly Lewinella sp., Haliscomenobacter sp., Runella sp.) and Nitrospirae were detected in sludge adapted to IL compared to the control system. The results showed that activated sludge can adapt to IL present in wastewater.

Biodegradability of industrial textile wastewater - batch tests.

Following new trends we applied oxygen uptake rate (OUR) tests as well as long-term tests (in two batch bioreactors systems) in order to assess the biodegradability of textile wastewater. Effluents coming from a dyeing factory were divided into two streams which differed in inorganic and organic contaminants loads. Usefulness of the stream division was proved. Biodegradation of the low-loaded stream led to over 97% reduction of biochemical oxygen demand (BOD5) together with 80% reduction of chemical oxygen demand (COD) and total organic carbon (TOC). Most of the controlled parameter values were below the levels allowed by legislation for influents to surface water, whereas the high-loaded stream was so contaminated with recalcitrant organic compounds that despite the reduction of BOD5 by over 95%, COD, TOC, total nitrogen and total phosphorus levels exceeded permissible values. OUR tests were aimed at determination of the following kinetic parameters: maximum specific growth rate (µMax), half-saturation constant, hydrolysis constant and decay coefficient for activated sludge biomass for both types of textile wastewater studied. The values of kinetic parameters will be applied in activated sludge models used for prediction and optimisation of biological treatment of textile wastewater.

Biodegradation of imidazolium ionic liquids by activated sludge microorganisms.

Biological properties of ionic liquids (ILs) have been usually tested with the help of standard biodegradation or ecotoxicity tests. So far, several articles on the identification of intermediate metabolites of microbiological decay of ILs have been published. Simultaneously, the number of novel ILs with unrecognized characteristics regarding biodegradability and effect on organisms and environment is still increasing. In this work, seven imidazolium ionic liquids of different chemical structure were studied. Three of them are 1-alkyl-3-methyl-imidazolium bromides, while the other four are tetra- or completely substituted imidazolium iodides. This study focused on the identification of intermediate metabolites of the aforementioned ionic liquids subjected to biodegradation in a laboratory activated sludge system. Both fully substituted ionic liquids and 1-ethyl-3-methyl-imidazolium bromide were barely biodegradable. In the case of two of them, no biotransformation products were detected. The elongation of the alkyl side chain made the IL more susceptible for microbiological decomposition. 1-Decyl-3-methyl-imidazolium bromide was biotransformed most easily. Its primary biodegradation up to 100 % could be achieved. Nevertheless, the cleavage of the imidazolium ring has not been observed.

Variations of morphology of activated sludge flocs studied at full-scale wastewater treatment plants.

Digital image analysis has been intensively developed over the last two decades including its application to describe morphology of activated sludge flocs. However, only few studies concerned the variation of flocs morphology with respect to the operational conditions, particularly oxido-reductive conditions, in a full-scale wastewater treatment plant (WWTP). In this work, morphology of activated sludge flocs was monitored over one year in two different full-scale WWTPs. The main aim of this study was to find the relationship between the operational parameters and morphology of sludge flocs. Simultaneously, the variations in floc size along activated sludge chamber were studied with respect to the oxido-reductive conditions. It was found that the sludge loading rate was one of the most important operational parameters influencing floc size. It was estimated that its values higher than 0.1 kg BOD5 kg TS(-1) d(-1) contributed to the decrease in floc size. Also, the oxido-reductive conditions influenced the floc size. It was statistically proved that flocs from the anaerobic zone were usually smaller than flocs from the anoxic or aerobic zones. Distribution of floc size in a full-scale WWTP usually could be described by a log-normal model.

Validation of a new image analysis procedure for quantifying filamentous bacteria in activated sludge.

Quantification of filamentous bacteria in activated sludge systems can be made by manual counting under a microscope or by the application of various automated image analysis procedures. The latter has been significantly developed in the last two decades. In this work a new method based upon automated image analysis techniques was elaborated and presented. It consisted of three stages: (a) Neisser staining, (b) grabbing of microscopic images, and (c) digital image processing and analysis. This automated image analysis procedure possessed the features of novelty. It simultaneously delivered data about aggregates and filaments in an individual calculation routine, which is seldom met in the procedures described in the literature so far. What is more important, the macroprogram performing image processing and calculation of morphological parameters was written in the same software which was used for grabbing of images. Previously published procedures required using two different types of software, one for image grabbing and another one for image processing and analysis. Application of this new procedure for the quantification of filamentous bacteria in the full-scale as well as laboratory activated sludge systems proved that it was simple, fast and delivered reliable results.

Comparison of imidazolium ionic liquids and traditional organic solvents: effect on activated sludge processes.

Data concerning the biodegradability and ecotoxicity of ionic liquids (ILs) obtained so far are insufficient in the context of IL removal from wastewater in activated sludge systems. Thus, in this work the selected imidazolium ionic liquids and two organic solvents (methanol and acetone) were tested with respect to their influence on activated sludge processes, particularly on the morphology of sludge flocs. The presence of ionic liquids with the chemical structure of 1-alkyl-3-methyl imidazolium bromide in wastewater did not deteriorate biological wastewater treatment processes if their concentration was not higher than 5 mg l(-1). Regarding the structure of the ILs studied, the longer the alkyl substituent was, the stronger the effect on sludge flocs. The highest decrease in activated sludge floc area and biomass concentration was exerted by the ionic liquid with the longest alkyl chain, i.e. 1-decyl-3-methylimidazolium bromide. The action of both methanol and acetone on floc size, activated sludge concentration and efficiency of organic pollutants removal was weaker compared to all tested 1-alkyl-3-methyl imidazolium bromides.

Effect of Innovative Bio-Based Plastics on Early Growth of Higher Plants.

Plastic particles are widespread in the environment including the terrestrial ecosystems. They may change the physicochemical properties of soil and subsequently affect plant growth. In recent decades, traditional, petroleum-derived plastics have been increasingly replaced by more environmentally friendly bio-based plastics. Due to the growing role of bio-based plastics it is necessary to thoroughly study their impact on the biotic part of ecosystems. This work aimed for the assessment of the effect of five innovative bio-based plastics of different chemical composition and application on the early growth of higher plants (sorghum, cress and mustard). Each bio-based plastic was tested individually. It was found that the early stages of growth of monocotyledonous plants were usually not affected by any of plastic materials studied. At the same time, the presence of some kinds of bio-based plastics contributed to the inhibition of root growth and stimulation of shoot growth of dicotyledonous plants. Two PLA-based plastics inhibited root growth of dicotyledonous plants more strongly than other plastic materials; however, the reduction of root length did not exceed 22% compared to the control runs. PBS-based plastic contributed to the stimulation of shoot growth of higher plants (sorghum, cress and mustard) at the concentrations from 0.02 to 0.095% w/w. In the case of cress shoots exposed to this plastic the hormetic effect was observed. Lepidium sativum turned out to be the most sensitive plant to the presence of bio-based plastic particles in the soil. Thus, it should be included in the assessment of the effect of bio-based plastics on plant growth.

Effect of bio-based microplastics on earthworms Eisenia andrei.

The contribution of bio-based plastics in the global market is gradually growing and diversifying. Therefore, it is necessary to assess their environmental impact including the biotic parts of ecosystems. Earthworms are regarded as functionally essential and useful bioindicators of ecological disturbances in the terrestrial ecosystems. The purpose of this study was to evaluate the impact of three innovative bio-based plastics on earthworms Eisenia andrei in the long-term experiments. It comprised the mortality, body mass and reproduction ability of earthworms as well as the oxidative stress response. Regarding the latter the activities of catalase (CAT) and superoxide dismutase (SOD) involved in the antioxidant system of earthworms were determined. Two out of three bio-based materials tested were polylactic acid-based (PLA-based) plastics, while one was poly(hydroxybutyrate-co-valerate)-based (PHBV-based) plastic. Neither mortality nor weight of adult earthworms was affected even at high concentration of the bio-based plastics up to 12.5 % w/w in the soil. Reproduction ability occurred to be more sensitive endpoint than mortality or body mass. At the concentration of 12.5 % w/w each of the studied bio-based plastics contributed to the decrease of the earthworm reproduction at statistically significant level. PLA-based plastics exerted stronger effect on earthworm reproduction ability than PHBV-based plastic did. CAT activity turned out to be a good indicator of the cellular response against oxidative stress induced by bio-based plastics in earthworms. The activity of this enzyme increased in the response to the exposure to the bio-based plastics compared to the level achieved in the control tests. It was from 16 % to about 84 % dependent on the material tested and its concentration in the soil. Finally, the reproduction ability and catalase activity are recommended to be used in the evaluation of the potential impacts of bio-based plastics on earthworms.

Application of a small scale-terrestrial model ecosystem (STME) for assessment of ecotoxicity of bio-based plastics.

In this work a small-scale terrestrial model ecosystem (STME) filled with 4 kg of soil mimicking the natural ecosystem was constructed, and then successfully applied to evaluate the effect of bio-based plastics on soil biota at the community level. It was inhabited by higher plants monocotyledonous (Sorghum saccharatum) and dicotyledonous (Lepidium sativum), and earthworms (Eisenia andrei). Two innovative bioplastics based on polylactic acid (PLA) were tested. This work is one of the first studies, in which potential impacts of bioplastic particles on soil organisms were determined at the community level. Owing to the application of the STME the ecotoxicity data for plants and earthworms were simultaneously collected and the mutual interactions might be taken into account. PLA-based plastics studied did not affect the percentage of seed germination of higher plants that was on average not lower than 88.9%. Neither the length nor fresh mass of shoots of cress were affected. One out of two PLA-based plastics (BPE-RP-PLA) inhibited sorghum growth so that it was statistically significant. PLA-based plastics did not cause to the mortality of earthworms as all ten organisms introduced to each STME survived each experiment. However, the presence of PLA-based plastic particles influenced the depth distribution of earthworms in the STMEs. Most of earthworms (60-70%) exposed to PLA-based plastics migrated downwards to the bottom soil zone, while 80% of the earthworms not exposed to PLA-based plastics lived in the top soil zone of the STME. This avoidance behaviour of earthworms known earlier for other contaminants (e.g. metals, pesticides) was for the first time reported with regard to bioplastic particles present in soil. It is a dangerous phenomenon not only for earthworms but also for the functioning and structure of terrestrial ecosystems. The STME proved to be an appropriate tool to detect it.

Phytotoxicity assessment of biodegradable and non-biodegradable plastics using seed germination and early growth tests.

Global production of plastics remains at the high level despite the SARS-Cov-2 pandemic. These are primarily petroleum-derived plastics but the contribution of bio-based plastics estimated at the level of 1% in the plastic market in 2019 is expected to be increasing. Simultaneously, the significant part of plastic waste is still disposed improperly and pollutes the environment making a threat to all living organisms. In this work three plastic materials, two bio-based biodegradable: polylactide (PLA) and polyhydroxybutyrate (PHB), and one petroleum-derived non-biodegradable polypropylene (PP) were studied towards their effects on seed germination and early growth of higher plants. The following plants were used as bioindicators: monocotyledonous plant - Sorghum saccharatum and two dicotyledonous plants: Sinapsis alba and Lepidium sativum. Plastics did not affect seed germination of higher plants even at the highest concentration tested (11.9% w/w) but their presence in soil acted in various ways on growth of the plants. Either no or inhibitive or stimulation effects on growth of roots or stems were noticed. It depended on the concentration and chemical composition of the plastic tested, and plant species. PHB and PLA more often caused to the inhibition of root growth than PP did. This phenomenon was observed in particular with regard to the dicotyledonous plants. Moreover, in the tests with the dicotyledonous plants (S. alba and L. sativum) the dose-response relations were usually determined as statistically relevant. Among these plants cress (L. sativum) occurred to be more sensitive and allowed for obtaining the dose-response dependence for both root and stem length, and, what is important, it took place in the case of each of materials tested. Therefore, cress is recommended to be used as a bioindicator in the assessment of the effect of plastics (petroleum-derived and bio-based plastics) on the early stages of growth of higher plants.

Calibration of a complex activated sludge model for the full-scale wastewater treatment plant.

In this study, the results of the calibration of the complex activated sludge model implemented in BioWin software for the full-scale wastewater treatment plant are presented. Within the calibration of the model, sensitivity analysis of its parameters and the fractions of carbonaceous substrate were performed. In the steady-state and dynamic calibrations, a successful agreement between the measured and simulated values of the output variables was achieved. Sensitivity analysis revealed that upon the calculations of normalized sensitivity coefficient (S(i,j)) 17 (steady-state) or 19 (dynamic conditions) kinetic and stoichiometric parameters are sensitive. Most of them are associated with growth and decay of ordinary heterotrophic organisms and phosphorus accumulating organisms. The rankings of ten most sensitive parameters established on the basis of the calculations of the mean square sensitivity measure (δ(msqr)j) indicate that irrespective of the fact, whether the steady-state or dynamic calibration was performed, there is an agreement in the sensitivity of parameters.

Effect of (bio)plastics on soil environment: A review.

The contribution of improperly disposed plastic wastes is globally evaluated at the level of 30% and these wastes make a particular threat to all living creatures. Thus, the evaluation of the possible impacts of plastic particles on the biotic part of ecosystems has become increasingly important in recent years. As a result, the growing number of publications concerning this subject has been observed since 2018. This paper aims to review the advances in studies on the effect of petroleum-derived plastic and bioplastic particles, taken together in the term (bio)plastics, on the terrestrial ecosystem, particularly on soil biota. It is the first review, in which both petroleum-derived plastics and bioplastics were analysed regarding their potential impacts on the soil compartment. Petroleum-derived plastics were more frequently studied than bioplastics and among analysed papers about 18% concern bioplastics. It was found that (bio)plastics did not affect the germination of seeds. However, they might contribute to the delay in germination processes. Both inhibitory and stimulating effects were observed in relation to the growth of roots and stems. (Bio)plastic microparticles did not inhibit the biochemical activity of nitrifiers and transformation of carbon compounds. Earthworms were predominantly used organisms to test the effect of petroleum-derived plastics on soil biota but there are hardly any data about bioplastics. Petroleum-derived microplastics present in soil at concentrations up to 1000 mg kg-1 usually neither cause to the mortality of earthworms nor affect their reproduction. Micro- and nanoparticles of petroleum-derived plastics could be accumulated in the earthworm intestine and transferred in the food chain. Summarizing, a high variability of results and often appearing lack of dose-dependence relationships hamper the final evaluation of the ecotoxicity of (bio)plastics simultaneously creating a need to develop the ecotoxicological studies on (bio)plastics, especially including these on the effect of bioplastics on soil animals.

Effect of anionic and nonionic surfactants on the kinetics of the aerobic heterotrophic biodegradation of organic matter in industrial wastewater.

While using the contemporary mathematical models for activated sludge systems, it is necessary to describe quantitatively the kinetics of microbiological processes and to characterise substrate (wastewater components in the influent) as well as biomass (activated sludge). In this paper, the kinetic parameters of the aerobic biodegradation of organic matter in wastewater containing synthetic surfactants in an activated sludge system were determined and discussed. Also, the composition of the tested wastewater was estimated and expressed as COD fractions. Five synthetic surfactants, three anionic and two nonionic, of different chemical structure were investigated. Each of them was tested separately and dissolved in wastewater to obtain a concentration of 50 mgl(-1), which can be found in some industrial wastewater. The presence of the elevated amount of synthetic surfactants in wastewater decreased the affinity of biomass to substrate. Nevertheless, maximum specific growth rates (micromax) of heterotrophic biomass exposed to wastewater containing surfactants were high but usually lower than micromax estimated for wastewater without surfactant. Surfactants, which contain a benzene ring, were the most likely to deteriorate wastewater treatment processes in the activated sludge systems.

Kinetics of Biological Removal of the Selected Micropollutants and Their Effect on Activated Sludge Biomass.

17α-Ethinylestradiol (EE2), diclofenac (DCF), and 4-nonylphenol (4NP) belong to the most common micropollutants (MPs) occurring in municipal wastewater treatment plants (WWTPs). The WWTPs are the primary barrier against the spread of micropollutants in the environment. The aim of this work was to study the kinetics of biological removal of the three aforementioned micropollutants from wastewater and to check whether the acclimation of biomass influenced on the kinetic parameters. In addition, the effect of MPs on the biochemical activity of microorganisms was tested. DCF inhibited the respiration activity of biomass to the highest extent, followed by 4NP and EE2, respectively. DCF occurred to be less susceptible to microbial decomposition than the other two MPs and was removed from wastewater at the lowest degree of 58%. The degrees of removal of EE2 and 4NP were higher than that of DCF and equal to 93 and 71%, respectively. The kinetic parameters determined in this work can be used in modelling and simulation of the removal of micropollutants from wastewater. They improve the predictive ability of the biokinetic models. The acclimation of the biomass to the relevant micropollutant does not influence on the kinetic parameters of biomass growth; however, it causes the increase of the yield coefficient for heterotrophic biomass.

Evaluation of biological properties and fate in the environment of a new class of biosurfactants.

Selected sophorolipid quaternary ammonium salts (SQAS), being a new class of modified biosurfactants, were studied in this work for the first time with regard to their biodegradability and fate in the environment. It was made to find whether environment-friendly bioproducts like biosurfactants are still safe to the environment after their chemical modification. The susceptibility of these SQAS for biodegradation was estimated together with the evaluation of their influence on activated sludge microorganisms. Additionally, the mechanisms of removal of the SQAS from wastewater and from the aquatic environment, were analysed. The evaluated SQAS were potentially biodegradable, although none of them could be classified as readily biodegradable. The biodegradation degrees after 28 days ranged from 4 to 42%, dependent on the SQAS tested, i.e. below the required OECD 301D Closed Bottle Test level of 60%. Simultaneously, the analysis of the mass spectra revealed the presence of the breakdown products of each SQAS studied. Biodegradation was preceded by sorption of the SQAS on sludge particles, which occurred to be a main mechanism of the removal of these newly synthesized biosurfactants from wastewater. The mean degree of sorption calculated on the basis of SQAS determination was from 75 to 96%, dependent on the studied SQAS. The presence of SQAS in wastewater did not deteriorate the operation of the activated sludge system, although the products of the SQAS biodegradation remained in the liquid phase and might contribute to the increase of COD of the effluent to be introduced to the environment.

Study on wastewater toxicity using ToxTrak™ method.

ToxTrak™ method is an analytical tool for the measurement of toxicity of drinking water, wastewater and natural water. It is based upon the estimation of the inhibitive effect on bacterial respiration processes. The main aim of this work was to test the applicability of ToxTrak™ method in the assessment of wastewater toxicity in a full-scale WWTP in Poland. In order to achieve it, the study was divided into two parts. First, the validation of ToxTrak™ method was performed. Second, wastewater toxicity was monitored in the long- and short-term campaigns. Validation of ToxTrak™ method revealed that the indigenous biomass (mixed cultures of activated sludge microorganisms) was more sensitive than Escherichia coli for both materials (wastewater and phenol) tested. The values of degree of inhibition determined for phenol towards indigenous biomass and E. coli were close to each other, and no statistically significant difference between them was found. It confirmed the reliability of the results obtained with the help of ToxTrak™ test. The toxicity of the effluent was always lower than that of the influent and the linear correlation between them was found. Despite, the decrease of wastewater toxicity in the WWTP, the effluents were ranked as toxic or highly toxic according to the classification of wastewater based upon the acute toxicity.

Acute toxicity and genotoxicity of five selected anionic and nonionic surfactants.

The results of four toxicity bioassays of selected anionic and nonionic surface active agents were presented. Three widely used anionic surfactants that belong to alkyl sulphates (AS), alkylbenzene sulphonates (LAS) and alkylpolyoxyethylene sulphates (AES) as well as nonionic surfactants: polyoxyethylene alkyl ethers (AE) and polyoxylethylene alkylphenyl ethers (APE) were tested. Three different toxicity assays to aquatic organisms: Physa acuta Draparnaud, Artemia salina and Raphidocelis subcapitata were applied. Additionally, the genotoxicity test with Bacillus subtilis M45 Rec- and H17 Rec+ strains was performed. The obtained results showed that none of the surfactants studied was genotoxic at the concentration 1000 mg l(-1). On the basis of toxicity tests to aquatic organisms all tested anionic surfactants were harmful (LC50 between 10 and 100 mg l(-1)), whereas nonionic ones were toxic (LC50 between 1 and 10 mg l(-1)) or even highly toxic (LC50 below 1 mg l(-1)). Moreover, the bigger was the molecular weight of the tested compound, the higher toxicity was observed.

Estimation of viable biomass in aerobic biodegradation processes of organic fraction of municipal solid waste (MSW).

2-(p-Iodophenyl)-3-(p-nitrophenyl)-5-phenyltetrazolium chloride (INT) dehydrogenase test and RNA assay were introduced to evaluate biomass in the processes of aerobic biodegradation of the organic fraction of municipal solid waste (MSW) in bioreactors. It was found that RNA quantification by KOH/UV method delivered reliable and repeatable results. Relative standard deviation (RSD) for INT test was significantly higher than for RNA assay and achieved values of 3-15%. Moreover, it occurred that the optimum temperature for the growth of autochthonic biomass, which takes part in the biodegradation process, was in the range from 25 to 37 degrees C.