Biological effects of vanillic acid, iso-vanillic acid, and orto-vanillic acid as environmental pollutants.

Vanillic acid (4-hydroxy-3-methoxybenzoic acid) (VA) is a natural benzoic acid derivative commonly found in herbs, rice, maize, and some fruits and vegetables. However, due to the wide use of VA in various industrial sectors, its presence in the environment might harm living organisms. This study evaluated the toxicity of VA and its isomers, iso-VA and orto-VA. Firstly, the antimicrobial effect of VA and its isomers iso-VA and orto-VA (in doses of 1000; 100, 10, 1; 0.1; 0.01 mg/L) against Escherichia coli, Sarcina spp., Enterobacter homaechei, Staphylococcus aureus and Candida albicans were identified. The toxic effect and protein degradation potential of VA and its isomers were determined using E. coli grpE:luxCDABE and lac:luxCDABE biosensor strains. However, the genotoxicity and oxidative stress generation were assessed with the E. coli recA:luxCDABE biosensor and E. coli strain. The results showed that VA, iso-VA, and orto-VA exhibited antimicrobial activity against all tested bacterial strains. However, VA's antimicrobial effect differed from iso-VA and orto-VA. Similar toxic, genotoxic, and oxidative stress-inducing effects were observed for VA and its isomers. Each compound exhibited toxicity, cellular protein degradation, and genotoxic activity against E. coli grpE:luxCDABE, E. coli lac:luxCDABE, and E. coli recA:luxCDABE strains. Analysis of reactive oxygen species (ROS) generation within E. coli cells highlighted oxidative stress as a contributing factor to the toxicity and genotoxicity of VA and its isomers. While the findings suggest potential applications of VA compounds as food preservatives, their presence in the environment raises concerns regarding the risks posed to living organisms due to their toxic and genotoxic characteristics.

Determination of the Effect of Wastewater on the Biological Activity of Mixtures of Fluoxetine and Its Metabolite Norfluoxetine with Nalidixic and Caffeic Acids with Use of E. coli Microbial Bioindicator Strains.

In the present work, the conducted research concerned the determination of the toxicity and oxidative stress generation of the antidepressant fluoxetine (FLU), its metabolite nor-fluoxetine (Nor-FLU), the antibiotic nalidixic acid (NA), caffeic acid (CA) and their mixtures in three different environments: microbial medium (MM), raw wastewaters (RW) and treated wastewaters (TW). We evaluated the following parameters: E. coli cell viability, toxicity and protein damage, sodA promoter induction and ROS generation. It was found that FLU, Nor-FLU, NA, CA and their mixtures are toxic and they have the potency to generate oxidative stress in E. coli strains. We also detected that the wastewater, in comparison to the microbial medium, had an influence on the toxic activity and oxidative stress synthesis of the tested chemicals and their mixtures. Regardless of the environment under study, the strongest toxic activity and oxidative stress generation were detected after bacterial incubation with NA at a concentration of 1 mg/dm3 and the mixture of FLU (1 mg/dm3) with Nor-FLU (0.1 mg/dm3) and with NA (0.1 mg/dm3). The ROS synthesis and sodA promoter induction suggest that, in the case of the examined compounds and their mixtures, oxidative stress is the mechanism of toxicity. The analysis of the types of interactions among the substances constituting the mixtures in the wastewater revealed synergism, potentiation and antagonism.

The study of biological activity of mandelic acid and its alkali metal salts in wastewaters.

In the present work we compared the biological activity of mandelic acid (MA) and its Li, Na, K, Rb and Cs salts. The study also investigated the effect of raw wastewaters (RW) and treated wastewaters (TW), comparable to microbial medium (MM) on the biological activity of the tested chemical compounds used in concentrations of 5; 2.5; 1.25; 0.625; 0.3125 mg/ml. In the present experiment the evaluation of the following parameters was performed: E. coli (ATCC 25922) cells viability, growth inhibition of E. coli (ATCC 25922), the inhibition of GFP protein, genotoxicity and ROS generation. Our results showed that three main factors differentiated the antibacterial activity of MA and its Li, Na, K, Rb and Cs salts: study environment (MM, RW, TW), metal forming salt of mandelic acid and concentration of tested compounds. Additionally, raw and treated wastewater, compared to microbial medium, changes the antimicrobial activity of MA and its salts in relation to the E. coli strain. We also detected that both MA and its salts affect the GFP protein and the induction of the recA promoter (genotoxicity test). The activity of the tested salts in relation to these two parameters is strictly dependent on the type of salt-forming metal and the concentration used. The analysis of ROS synthesis suggests that in the majority of the studied mandelic acid salts, oxidative stress is the dominant mechanism of cytotoxicity and genotoxicity. We also showed that both raw wastewaters (RW) and treated wastewaters (TW), compared to microbial medium (MM), change significantly the activity of MA and its salts.

Natural Cinnamic Acid Derivatives: A Comprehensive Study on Structural, Anti/Pro-Oxidant, and Environmental Impacts.

Cinnamic acid (CA), p-coumaric acid (4-hydroxycinnamic acid, 4-HCA), caffeic acid (3,4-vdihydroxycinnamic acid, 3,4-dHCA), and 3,4,5-trihydroxycinnamic acid (3,4,5-tHCA) were studied for their structural, anti-/pro-oxidant properties and biodegradability. The FT-IR, FT-Raman, UV/Vis, 1H and 13C NMR, and quantum chemical calculations in B3LYP/6-311++G** were performed to study the effect on number and position of hydroxyl group in the ring on the molecular structure of molecules. The antioxidant properties of the derivatives were examined using DPPH● and HO● radicals scavenging assays, ferric ion reducing antioxidant power (FRAP), cupric reducing antioxidant capacity (CUPRAC), inhibition of linoleic acid oxidation, as well as the biological antioxidant assay with Saccharomyces cerevisiae. Moreover, the pro-oxidant activity of compounds in Trolox oxidation assay was estimated. The effect of the derivatives on environment on the basis of increasing the carbon and nitrogen compounds transformation processes occurring in biological wastewater treatment was studied.

Biosorption of LMW PAHs on activated sludge aerobic granules under varying BOD loading rate conditions.

Polycyclic aromatic hydrocarbons belong to the main priority substances for the aquatic environment. One of the emission sources of these compounds to environment is wastewater discharged from conventional wastewater treatment systems, which are not designed to cope with this type of pollution. Thus, due to the widely discussed properties of aerobic granular activated sludge in the literature - a conducted study has proven its ability to remove LMW PAHs (naphthalene (Nap), acenaphthylene (Acy), acenaphthene (Ace), fluorene (Flu), phenanthrene (Phe) and anthracene (Ant)) from wastewater by biosorption process at varying loadings of organic compounds expressed as BOD (kg/kg·d) on the activated sludge mass. The maximum biosorption of Nap was 605 µg/kgd.m., Acy equals to 134 µg/kgd.m., Ace equals to 355 µg/kgd.m. Flu equals to 104 µg/kgd.m. Phe equal to 204 µg/kgd.m. and Ant equal to 173 µg/kgd.m. The study showed that the BOD loading rate is one of the factors affecting the biosorption process of LMW PAHs. However, as the amount of adsorbed LMW PAHs increased, the condition of aerobic granular activated sludge deteriorated, which was evidenced by gradual increase in the values of technological parameters of activated sludge (SVI, HRT, SRT) and a smaller increase in activated sludge dry mass.

Synergistic interaction of diclofenac and its metabolites with selected antibiotics and amygdalin in wastewaters.

Diclofenac (DCF), a non-steroidal anti-inflammatory drug (NSAID) belongs to one of the most frequently detected pharmaceutical residues in the environment. Little is known on the interactions of DCF as well as its major biodegradation metabolites 4'-OHDCF and 5-OHDCF with chemical compounds found in wastewater, including antibiotics such as ampicillin and kanamycin. In the present work we examined the potential interactions between DCF, its metabolites 4'-OHDCF and 5-OHDCF and ampicyllin and kanamycin. We also measured the effect of the mixture of DCF with natural compound - amygdalin. We evaluated the following parameters: E. coli K-12 cells viability, growth inhibition of E. coli K-12 culture, genotoxicity, oxidative stress parameters: sodA promoter induction and ROS generation. The reactivity of E. coli SM recA:luxCDABE biosensor strain in wastewaters matrices contaminated with DCF and kanamycin was also monitored. Obtained results indicated that used antibiotics (ampicyllin, kanamycin) enhanced the toxic effect of DCF used individually and in the mixtures with its metabolites 4'-OHDCF and 5-OHDCF toward E. coli. Similar effect was also obtained in genotoxicity assay. The oxidative stress assays revealed that the highest level of ROS generation and sodA promoter induction were obtained also for the mixtures of DCF, its metabolites with antibiotics. It was also showed that amygdalin influenced the activity of DCF and its biodegradation metabolites. The strongest luminescence response of E. coli SM biosensor strain with recA:luxCDABE genetic construct in filtered treated wastewaters, comparable to control sample was noticed. Obtained results showed that DCF and its biodegradation metabolites 4'-OHDCF and 5-OHDCF can interact with tested antibiotics and compounds of natural origin, i.e. amygdalin to form mixtures showing stronger antimicrobial activity against E. coli than parent chemicals. Moreover the assays in wastewater matrices revealed that E. coli SM recA:luxCDABE biosensor strains is a good tool for bacteria monitoring in wastewater environments.

The study of biological activity of transformation products of diclofenac and its interaction with chlorogenic acid.

In the present work we compared the biological activity of DCF, 4'-OHDCF and 5-OHDCF as molecules of most biodegradation pathways of DCF and selected transformation products (2-hydroxyphenylacetic acid; 2,5-dihydroxyphenylacetic acid and 2,6-dichloroaniline) which are produced during AOPs, such as ozonation and UV/H2O2. We also examined the interaction of DCF with chlorogenic acid (CGA). CGA is commonly used in human diet and entering the environment along with waste mainly from the processing and brewing of coffee and it can be toxic for microorganisms included in activated sludge. In the present experiment the evaluation of following parameters was performed: E. coli K-12 cells viability, growth inhibition of E. coli K-12 culture, LC50 and mortality of Chironomus aprilinus, genotoxicity, sodA promoter induction and ROS generation. In addition the reactivity of E. coli SM recA:luxCDABE biosensor strain in wastewater matrices was measured. The results showed the influence of DCF, 4'-OHDCF and 5-OHDCF on E. coli K-12 cells viability and bacteria growth, comparable to AOPs by-products. The highest toxicity was observed for selected, tested AOPs by-products, in comparison to the DCF, 4'-OHDCF and 5-OHDCF. Genotoxicity assay indicated that 2,6-dichloroaniline (AOPs by-product) had the highest toxic effect. The oxidative stress assays revealed that the highest level of ROS generation and sodA promoter induction were obtained for DCF, 4'-OHDCF and 5-OHDCF, compared to other tested compounds. We have also found that there is an interaction between chlorogenic acid and DCF, which resulted in increased toxicity of the mixture of the both compounds to E. coli K-12, comparable to parent chemicals. The strongest response of E. coli SM biosensor strain with recA:luxCDABE genetic construct in filtered treated wastewaters, comparable to control sample was noticed. It indicates, that E. coli SM recA:luxCDABE biosensor strains is a good tool for bacteria monitoring in wastewater environment. Due to toxicity and biological activity of tested DCF transformation products, there is a need to use additional wastewater treatment systems for wastewater contaminated with pharmaceutical residues.

Alternated biodegradation of naphthalene (NAP), acenaphthylene (ACY) and acenaphthene (ACE) in an aerobic granular sludge reactor (GSBR).

The paper presents quantitative changes of selected 2- and 3-ring PAHs after process phases of GSBR reactor. The studies have been carried out for 264 cycles of GSBR reactor, during which concentration of naphthalene was increased in the range of 3.00-710.00 µg/L, acenaphthylene 1.00-160.00 µg/L, acenaphthene 3.00-440.00 µg/L. GSBR operating cycle consisted of filling (30 min), mixing (90 min), aeration (540 min), sedimentation (10 min), decanting (30 min) and downtime (20 min) phases. Activated sludge dry mass concentration was 4.00 kg/m3. Conducted studies showed that in GSBR reactor naphthalene was degraded with the highest intensity. Results of the statistical analysis confirmed that naphthalene concentrations were statistically significantly different (α = 0.05) after each individual GSBR process phase, while in case of acenaphthene and acenaphthylene, the differences were observed only between mixing and aeration phases. Additionally, equations estimating concentrations of PAHs in treated wastewater were developed. Selected activated sludge technological parameters (sludge volume index, sludge and hydraulic retention time) and concentration of PAHs were used for equations. The R2 coefficients of equations were above 0.99, which indicates a good adjustment of estimation to observed values.