How to accurately assess surfactant biodegradation-impact of sorption on the validity of results.

Surfactants not only are widely used in biotechnological processes but also constitute significant contaminants of the modern world. Among many reports, there is a shortage of works which summarize the issue of surfactant sorption to biomass in a way that would elucidate the biological factors for analysts and analytical factors for microbiologists. The main factor, which is not as obvious as one would expect, is associated with the susceptibility of analytical approaches to errors resulting from incorrect handling of biomass. In case of several publications reviewed in the framework of this study, it was not possible to establish whether the decrease of the analytical signal observed by the authors actually resulted from biodegradation of the surfactant. This review emphasizes the necessity to consider the possibility of surfactant sorption to microbial cells, which may result in significant detection errors as well as conceptual inconsistency. In addition, a reference study regarding representative surfactants (cationic, anionic and non-ionic) as well as yeast, Gram-negative, Gram-positive bacteria, and activated sludge was provided to highlight the possible errors which may arise from disregarding sorption processes when determining degradation of surfactants. This particularly applies to systems which include ionic surfactants and activated sludge as sorption may account for 90% of the observed depletion of the surfactant. Therefore, a systematic approach was proposed in order to improve the credibility of the obtained results. Finally, the need to employ additional procedures was highlighted which may be required in order to verify that the decrease of surfactant concentration results from biodegradation processes.

Continuous Flow Methylene Blue Active Substances Method for the Determination of Anionic Surfactants in River Water and Biodegradation Test Samples.

Anionic surfactants are commonly determined with the use of the methylene blue active substances (MBAS) standard method, which is time-consuming and labor-intensive. Therefore, new methods for determination of anionic surfactants are needed. In this study, the standard MBAS method for determination of anionic surfactants was modified and adjusted to work in a continuous flow system combined with spectrophotometric measurement. The developed method was found to be satisfactory in terms of sensitivity and precision, with a short time of analysis. The quantification limit for anionic surfactants was at 16 µg L-1, with a relative standard deviation of 1.3 % for a model sample and 3.8 % for a river water sample. The results obtained for environmental samples were comparable to those obtained by using the standard MBAS method; however, the developed continuous flow method is faster, more sensitive and consumes smaller doses of chemical reagents.

Biodegradation of Triton X-100 and its primary metabolites by a bacterial community isolated from activated sludge.

A set of studies was carried using a continuous flow biodegradation unit in order to isolate a microbial community capable of efficient and complete utilization of octylphenol ethoxylates from activated sludge. Increasing concentrations of Triton X-100 (in the range of 1-1000 mg/l) were applied over a time period of 35 days in order to select microorganisms, which exhibit high tolerance towards this surfactant. The fate of the surfactant and its primary degradation products was assessed by HPLC/MS. It was observed that even small doses of the surfactant contributed to the disruption of the activated sludge, due to adsorption of primary Triton X-100 metabolites (octylphenol and short-chained ethoxylates) on the cells, although the long-chain octylphenol ethoxylates were efficiently degraded during the isolation process. The toxicity assessment of octylphenol as well as octylphenol di- and monoethoxylates towards activated sludge allowed for determination of EC50 values (8 and 55 mg/l, respectively). The identification of the residual microorganisms revealed the presence of Acinetobacter junii, Acinetobacter calcoaceticus, Aeromonas hydrophilia, Alcaligenes spp., Pseudomonas fluorescens and Sphingomonas capsulata. The isolated community exhibited a high resistance towards Triton X-100 and was capable of growth even at 10,000 mg/l, with the highest specific growth rate (0.47 h(-1)) observed at 4000 mg/l. Under aerobic conditions both octylphenol and the short-chained ethoxylates were completely degraded while no toxic effect towards the isolated bacterial community was observed.

Fluoride exposure and factors affecting dental caries in preschool children living in two areas with different natural levels of fluorides.

BACKGROUND: Successful oral health interventions must be based on the specific needs of the population that they serve. Evaluation of habits related to dental caries development and estimation of fluoride exposure in a target group of young patients helps to plan effective and safe caries prevention strategies. OBJECTIVES: The study aimed to evaluate factors affecting dental caries experience and sources of fluoride exposure in preschool children living in two areas: with optimal and low natural content of fluoride in drinking water. MATERIALS AND METHODS: The study included a group of 73 children of both sexes aged 4-7 years attending two kindergartens in Sroda Wielkopolska and Turek (Wielkopolska Voivodeship, Poland), where the content of fluoride in drinking water according to data obtained in the sanitary station ranged from 0.68 to 0.74 mg/L (optimal concentration of fluoride) and from 0.19 to 0.30 mg/L (low concentration of fluoride), respectively. Parents of patients completed a survey about diet, hygiene, and dental care, taking into account the child's fluoride exposure. The calibrated dentist assessed the oral health condition using a mirror, a CPI probe, and a headlamp. Oral hygiene was recorded using the Silness and Löe plaque index, caries experience by calculating the numbers of decayed, missing, and filled primary, and permanent teeth (dmf and DMF, respectively) while caries frequency by calculating the percentage of children with caries experience above 0. In order to assess the fluoride concentrations in urine and drinking water, parents were asked to provide a urine sample collected on fasting and a tap water sample. Fluoride concentrations were assessed using a 09-37 (MARAT) fluoride ion-selective electrode and a RAE 111 silver-chloride reference electrode. Statistical analysis was conducted using the data analysis software system Statistica (version 12, StatSoft, Inc. 2014), assuming a statistical significance level p < 0.05. RESULTS: No statistically significant differences were found between caries indices of the examined children in each kindergarten (p > 0.05). Urinary fluoride levels were higher in children who tended to swallow toothpaste or used fluoride rinses and positively correlated with fluoride concentrations in the drinking water. Dental caries experience in the examined children depended on the effectiveness and frequency of oral hygiene procedures and dietary habits. CONCLUSIONS: The strategy aimed at improving the oral health of the examined group of children should include accomplishing oral hygiene, promoting a non-cariogenic diet, and, finally, controlling fluoride exposure from at-home fluoride products. Caries prevention program ought to be adjusted to individual characteristics of each child, taking into consideration oral hygiene practices, dietary habits and total fluoride intake.

Biodegradation of diesel fuel by a microbial consortium in the presence of 1-alkoxymethyl-2-methyl-5-hydroxypyridinium chloride homologues.

Fast development of ionic liquids as gaining more and more attention valuable chemicals will undoubtedly lead to environmental pollution. New formulations and application of ionic liquids may result in contamination in the presence of hydrophobic compounds, such as petroleum mixtures. We hypothesize that in the presence of diesel fuel low-water-soluble ionic liquids may become more toxic to hydrocarbon-degrading microorganisms. In this study the influence of 1-alkoxymethyl-2-methyl-5-hydroxypyridinium chloride homologues (side-chain length from C(3) to C(18)) on biodegradation of diesel fuel by a bacterial consortium was investigated. Whereas test performed for the consortium cultivated on disodium succinate showed that toxicity of the investigated ionic liquids decreased with increase in side-chain length, only higher homologues (C(8)-C(18)) caused a decrease in diesel fuel biodegradation. As a result of exposure to toxic compounds also modification in cell surface hydrophobicity was observed (MATH). Disulphine blue active substances method was employed to determine partitioning index of ionic liquids between water and diesel fuel phase, which varied from 1.1 to 51% for C(3) and C(18) homologues, respectively. We conclude that in the presence of hydrocarbons acting as a solvent, the increased bioavailability of hydrophobic homologues is responsible for the decrease in biodegradation efficiency of diesel fuel.

Evaluation of surfactant removal efficiency in selected domestic wastewater treatment plants in Poland.

The aim of this study was to evaluate the work of a two types of household sewage treatment plant: wetland wastewater treatment plant (ORS type) and treatment plant of SBR type (SBR-K-6 type). Physicochemical analyses of selected pollution indices (BOD5, COD, total suspension, total phosphorus) and surfactants were carried out and compared with currently applicable values of such indexes according to the Regulation of the Minister of the Environment in Poland on the conditions to be met when discharging sewage into water or soil, and on the substances particularly harmful to the aquatic environment. The removal efficiency of organic compounds, expressed as COD and BOD5, reached the threshold of 90%, which is required in regulations. In contrast, the effects of removal of biogenic compounds were low - in case of total nitrogen the removal rate reached approx. 40% and the desired admissible concentration of 30 mg N/L was not achieved. The reduction efficiency of total suspended solids reached 57.0 and 59.6% for the ORS and SBR-K-6 type objects, respectively, and therefore the required threshold of minimum 90% was not reached. Anionic surfactants were removed by up to 98 and 88% in the ORS and SBR-K-6 type wastewater treatment plants, respectively. Lower removal efficiency was achieved in case on non-ionic surfactants, which reached 76% for the ORS type object and 56% for the SBR-K-6 type object. This article proven high wastewater treatment efficiency and lower than necessary concentrations in the effluent from domestic wastewater treatment plants may be achieved mainly by proper exploitation of the devices and appropriately selected vegetation.

Effects of ammonium-based ionic liquids and 2,4-dichlorophenol on the phospholipid fatty acid composition of zebrafish embryos.

Ionic liquids consisting of a combination of herbicidal anions with a quaternary ammonium cation act as efficient herbicides, which are under consideration to be used in the agriculture. In the present study, we used embryos of the zebrafish (Danio rerio) as a model to assess the toxic potential of ammonium-based ionic liquids for aquatic organisms. As we assumed interference of the partially hydrophobic ionic liquid cation with lipids, we investigated the adaptation response in the lipid composition of the zebrafish embryos, triggered by the ionic compound. Therefore, the impact of ammonium-based ionic liquids with different lengths of the alkyl chain ([C6,C6,C1,C1N][Br], [C8,C8,C1,C1N][Br]) on the phospholipid fatty acid (PLFA) profile of zebrafish embryos up to 72 hours post fertilization (hpf) was examined. Furthermore, the changes in the unsaturation index (UI) of PLFAs, as the sum parameter of membrane fluidity in eukaryotic cells, were presented. The PLFA's UI in the zebrafish embryos upon exposure to quaternary ammonium salts was compared to the UI of the embryos upon exposure to nonionic 2,4-dichlorophenol, which has a similar hydrophobicity but is structurally different to [C8,C8,C1,C1N][Br]. It was shown that for ammonium-based ionic liquid precursors non-specific mode of action occurs and the toxic effect on lipid composition of zebrafish embryos can be well predicted based on chemical properties, like hydrophobicity. Furthermore, the changes in PLFAs, expressed by the UI, can be useful to study toxic effects of organic contamination. However, for zebrafish embryos, after ionic liquids and 2,4-DCP exposure, the changes were observed at high lethal concentrations, which caused the incidence of lethality of 30 and 50% of a group of test animals.

Toxicity evaluation of selected ammonium-based ionic liquid forms with MCPP and dicamba moieties on Pseudomonas putida.

Combination of the hydrophilic herbicidal anion with hydrophobic, antimicrobial ammonium cation allows to obtain compounds in ionic liquid form with better properties then conventional herbicides. Both cation and anion can be modified by selection of herbicide and the length of alkyl chains in cation structure. However the knowledge of their potential toxic effects are still limited. Furthermore, the relation between hydrophobicity associated with the length of alkyl chains and toxicity for ionic liquids has not been thoroughly studied. Therefore we investigated toxic effects of herbicidal ionic liquid forms on growth inhibition, given as EC50, of the common soil bacterium Pseudomonas putida. We thereby concentrated on quaternary ammonium salts. Analyzed compounds were composed of dicamba or MCPP moieties and cation with various alkyl chain lengths (n = 6,8,10) We compared them with commercial herbicides, and ammonium-based ionic liquids with neutral anion (Br-). In addition, cis-trans isomerisation of unsaturated membrane fatty acids in Pseudomonas putida was applied as the proxy for toxicity and membrane activity. We showed that toxicity increased with the length of alkyl chains. However, this correlation is only valid for six and eight carbon atom in alkyl chains, where for n = 10 the EC50 values rise by one order of magnitude. In our studies, the herbicidal ionic liquids [C10,C10,C1,C1N][MCPP] and [C10,C10,C1,C1N][dicamba] showed the lowest toxicity among analyzed quaternary ammonium salts and comparable toxicity with corresponding herbicides. No clear increase in toxicity could be followed by changing the anion moieties for ammonium-based ionic liquid forms.

Determination of non-ionic surfactants and their biotransformation by-products adsorbed on alive activated sludge.

A procedure has been developed for the determination of non-ionic surfactants (NS) adsorbed on particles of alive and dead activated sludge. The procedure also enables the determination of adsorption of major biodegradation by-products: short-chained ethoxylates, long- and short-chained PEG. The basis of measurement is the determination of NS concentration in a slurry of activated sludge and in a solution phase. The difference between these two concentrations represents the NS adsorbed on activated sludge. Separation of NS and their biotransformation by-products from samples and then on narrower fractions was performed by a sequential liquid-liquid extraction and precipitation with modified Dragendorff reagent. The indirect tensammetric technique (ITT) was applied for the final determination. The developed method was checked using the example of the treatment of the surfactant C12E10 (oxyethylated fatty alcohol) (C12E10) in the continuous flow activated sludge facility. No statistically significant accumulation of C12E10 on the alive activated sludge was detected, probably because of faster C12E10 fission than its adsorption. However, significant adsorption of the short-chained ethoxylates (including free alcohol) on the alive activated sludge was found, as well as statistically significant adsorption of long- and short-chained PEG. The adsorption of surfactant C12E10 and its biodegradation by-products on dead activated sludge was found to be higher than the species adsorption on alive activated sludge.

Biodegradation of oxo-alcohol ethoxylates in the continuous flow activated sludge simulation test.

Biodegradation of two alpha-methyl branched oxo-alcohol ethoxylates (OAE) of different polydispersity: LIAL 125/14 BRD (LIALB) (broad M.W. distribution) and LIAL 125/14 NRD (LIALN) (narrow M.W. distribution), both having an average of 14 oxyethylene subunits (EO) and a C(12-15) alkyl moiety were tested under the continuous flow activated sludge conditions of the classical Husmann plant. Primary biodegradation and concentration of metabolites: free oxo-alcohol fraction (FOA) and poly(ethylene glycols) (PEG), were measured. PEG were divided into two fractions: short-chained PEG (PEGshch) (1-4 EO) and long-chained PEG (PEGlch) (>4 EO). The indirect tensammetric technique combined with an adequate separation was used for analysis. Central fission was found to be a highly dominating pathway, as is the case with fatty alcohol ethoxylates. OAE are highly primarily biodegraded (above 95%). High concentrations of FOA and PEG are formed. Once formed the PEGlch are further fragmented into the PEGshch. Free alcohol fraction compounds are biodegraded sooner when alkyl moiety is shorter. OAE polydispersity has an influence on the kinetics of biodegradation; PEG formed from LIALN are biodegraded slower and to a lower degree than those from LIALB.

Genetic and chemical analyzes of transformations in compost compounds during biodegradation of oiled bleaching earth with waste sludge.

Composting of oiled bleaching earth with waste sludge and corn straw was carried out to investigate the ability of microorganisms to synthesize biosurfactants that might decrease the surface tension of composts. Analytical results and changes in the surface tension suggest that biodegradation of fatty by-products was the consequence of emulsifying properties of higher fatty acids. The surface tension for isolates from all composting phases was between 37 and 43 mN m(-1). No substances synthesized by microorganisms that might be able to decrease the surface tension were detected in composts. Tensammetric, TLC and HPLC-MS results and changes in surface tension suggest that biodegradation of fatty by-products results from the emulsifying properties of higher fatty acids. A decrease in fatty content from 144 to 6 mg g(-1) dry matter was obtained.

Utilization of Triton X-100 and polyethylene glycols during surfactant-mediated biodegradation of diesel fuel.

The hypothesis regarding preferential biodegradation of surfactants applied for enhancement of microbial hydrocarbons degradation was studied. At first the microbial degradation of sole Triton X-100 by soil isolated hydrocarbon degrading bacterial consortium was confirmed under both full and limited aeration with nitrate as an electron acceptor. Triton X-100 (600 mg/l) was utilized twice as fast for aerobic conditions (t(1/2)=10.3h), compared to anaerobic conditions (t(1/2)=21.8h). HPLC/ESI-MS analysis revealed the preferential biodegradation trends in both components classes of commercial Triton X-100 (alkylphenol ethoxylates) as well as polyethylene glycols. The obtained results suggest that the observed changes in the degree of ethoxylation for polyethylene glycol homologues occurred as a consequence of the 'central fission' mechanism during Triton X-100 biodegradation. Subsequent experiments with Triton X-100 at approx. CMC concentration (150 mg/l) and diesel oil supported our initial hypothesis that the surfactant would become the preferred carbon source even for hydrocarbon degrading bacteria. Regardless of aeration regimes Triton X-100 was utilized within 48-72 h. Efficiency of diesel oil degradation was decreased in the presence of surfactant for aerobic conditions by approx. 25% reaching 60 instead of 80% noted for experiments without surfactant. No surfactant influence was observed for anaerobic conditions.