Environmental impact assessment of dicationic ionic liquids with ammonium-phosphonium cations and amino acid anions.

Progress in the development of biodegradable or biobased ionic liquids (ILs) has led to the design of green compounds for several applications. Herein, four biocompatible dicationic ionic liquids (DILs) with ammonium-phosphonium cations and amino acid anions were synthesized and investigated their environmental impact. The structures of the DILs were confirmed by spectral analyses (1H, 13C and 31P NMR). Furthermore, physicochemical properties such as density, viscosity and refractive index were determined. Water content, bromide content and solubility were thereafter determined as the parameters needed for further studies. Subsequently, their antifeedant activity towards economically important pests of grain in storage warehouses: the granary weevil, the confused flour beetle, and the khapra beetle was examined, showing the dependence on structure. Moreover, selected DILs were investigated for toxicity towards white mustard, Daphnia magna, and Artemia franciscana to specify the environmental impact. These studies were complemented by understand the biodegradation of DILs by bacterial communities derived from soil at the agricultural land. The result was DILs with limited environmental footprints that have great potential for further application studies.

Cannabidiol as a Modulator of the Development of Alcohol Tolerance in Rats.

The study aimed to explore in vivo the influence of cannabidiol (CBD) on the development of alcohol tolerance in rats. Rats were treated with ethanol (3.0 g/kg, i.p.) and CBD (20 mg/kg, p.o.) for nine successive days, and rectal body temperature, sedation (sleeping time), and blood alcohol concentration (BAC) were measured. In the prefrontal cortex, hippocampus, and striatum, the cannabinoid (CB1R and CB2R) and dopaminergic (DRD1, DRD2, DRD4, DRD5) receptors' mRNA level changes were analyzed using the quantitative RT-PCR method. CBD inhibited the development of tolerance to the hypothermic and sedative action of alcohol, coupled with BAC elevation. On a molecular level, the most pronounced effects of the CBD + ethanol interaction in the striatum were observed, where CBD reversed the downregulation of CB2R gene transcription caused by ethanol. For CB1R, DRD1, and DRD2 mRNAs, the CBD + ethanol interaction produced opposite effects than for CB2R ones. In turn, for the transcription of genes encoding dopaminergic receptors, the most potent effect of alcohol as CBD occurred in the hippocampus. However, the combined CBD and alcohol administration showed the same effect for each substance administered separately. Since tolerance is considered a prelude to drug addiction, obtained results allow us to emphasize the thesis that CBD can inhibit the development of alcohol dependence in rats.

Multi-faceted analysis of bacterial transformation of nitrofurantoin.

Excessive presence of antibiotics and their residues can be dangerous to the natural environment. To reduce this negative effect, efficient strategies to remove them from the ecosystem are required. This study aimed to explore the potential of bacterial strains to degrade nitrofurantoin (NFT). Single strains isolated from contaminated areas, namely Stenotrophomonas acidaminiphila N0B, Pseudomonas indoloxydans WB, and Serratia marcescens ODW152 were employed in this study. Degradation efficiency and dynamic changes within the cells during NFT biodegradation were investigated. For this purpose, atomic force microscopy, flow cytometry, zeta potential, and particle size distribution measurements were applied. Serratia marcescens ODW152 showed the highest performance in removal of NFT (96 % in 28 days). The AFM images revealed modifications of cell shape and surface structure induced by NFT. Zeta potential showed significant variations during biodegradation. Cultures exposed to NFT had a broader size distribution than the control cultures due to increased cells agglomeration or aggregation. 1-Aminohydantoin and semicarbazide were detected as nitrofurantoin biotransformation products. They showed increased cytotoxicity toward bacteria as determined by spectroscopy and flow cytometry. Results of this study suggest that nitrofurantoin biodegradation leads to formation of stable transformation products that significantly affect the physiology and structure of bacterial cells.

Potential negative effect of long-term exposure to nitrofurans on bacteria isolated from wastewater.

Nitrofurans are broad-spectrum bactericidal agents used in a large quantity for veterinary and human therapy. This study reports the long-term impact of two nitrofuran representatives, nitrofurantoin (NFT) and furaltadone (FTD) on the bacterial strains Sphingobacterium siyangense FTD2, Achromobacter pulmonis NFZ2, and Stenotrophomonas maltophilia FZD2, isolated from a full-scale wastewater treatment plant. Bacterial whole genome sequencing was used for preliminary strains characterization. The metabolomic, electrochemical, and culture methods were applied to understand changes in the bacterial strains after 12-month exposure to nitrofurans. The most significantly altered metabolic pathways were observed in amino acid and sugar metabolism, and aminoacyl-tRNA biosynthesis. Disrupted protein biosynthesis was measured in all strains treated with antibiotics. Prolonged exposure to NFT and FTD also triggered mutagenic effects, affected metabolic activity, and facilitated oxidative stress within the cells. Nitrofuran-induced oxidative stress was evidenced from an elevated activity of catalase and glutathione S-transferases. NFT and FTD elicited similar but not identical responses in all analyzed strains. The results obtained in this study provide new insights into the potential risks of the prolonged presence of antimicrobial compounds in the environment and contribute to a better understanding of the possible impacts of nitrofuran antibiotics on the bacterial cells.

Dynamics of microbial communities during biotransformation of nitrofurantoin.

The purpose of this research was to investigate the biodegradation of nitrofurantoin (NFT), a typical nitrofuran antibiotic of potential carcinogenic properties, by two microbial communities derived from distinct environmental niches - mountain stream (NW) and seaport water (SS). The collected environmental samples represent the reserve of the protected area with no human intervention and the contaminated area that concentrates intense human activities. The structure, composition, and diversity of the communities were analyzed at three timepoints during NFT biodegradation. Comamonadaceae (43.2%) and Pseudomonadaceae (19.6%) were the most abundant families in the initial NW sample. The top families in the initial SS sample included Aeromonadaceae (31.4%) and Vibrionaceae (25.3%). The proportion of the most abundant families in both consortia was remarkably reduced in all samples treated with NFT. The biodiversity significantly increased in both consortia treated with NFT suggesting that NFT significantly alters community structure in the aquatic systems. In this study, NFT removal efficiency and transformation products were also studied. The biodegradation rate decreased with the increasing initial NFT concentration. Biodegradation followed similar pathways for both consortia and led to the formation of transformation products: 1-aminohydantoin, semicarbazide (SEM), and hydrazine (HYD). SEM and HYD were detected for the first time as NFT biotransformation products. This study demonstrates that the structure of the microbial community may be directly correlated with the presence of NFT. Enchanced biodiversity of the microbial community does not have to be correlated with increase in functional capacity, such as the ability to biodegradation because higher biodiversity corresponded to lower biodegradation. Our findings provide new insights into the effect of NFT contamination on aquatic microbiomes. The study also increases our understanding of the environmental impact of nitrofuran residues and their biodegradation.

Azole fungicides: (Bio)degradation, transformation products and toxicity elucidation.

The increasing consumption of azole antifungal agents leads to their uncontrolled release into the environment. Therefore, it is crucial to remove their residues from natural ecosystems. This study aimed to examine the biological and chemical degradation of four typical azole fungicides: fluconazole (Fc), clotrimazole (Cl), climbazole (Cb), and epoxiconazole (Ep). The biodegradation was investigated using activated sludge and two novel Gram-negative bacterial strains. The chemical degradation experiments aimed to assess the efficiency of fungicides removal through UV treatment, the Fenton reaction, and a combination of these methods. Transformation products of Cb, Ep, and Cl photocatalytic removal were identified by mass spectrometry. In addition, the AlamarBlue® Assay and the MTT Assay allowed careful evaluation of the toxicity of azole derivatives and their transformation products towards newly isolated strains, Stenotrophomonas maltophilia AsPCl2.3 and Pseudomonas monteilii LB2. Among all azole fungicides, Cb was the most susceptible to biological removal while Fc, Ep, and Cl were basically resistant to biodegradation. Cl and Ep showed a significant biosorption on the activated sludge. Under optimized photolysis conditions, the removal efficiency of Cl, Cb, and Ep was significantly higher than that of biodegradation. The Fenton reaction supported by the UV-irradiation offered the best results of fungicides elimination. After 1 min of the experiment, Cl was almost completely removed while Cb and Ep removal rates reached an average of 60%. The proposed main degradation route of azole fungicides during UV-irradiation includes halogen atoms substitution by hydroxyl moieties. The final degradation product was imidazole or triazole. Azole fungicides and their transformation products differently affected the metabolic activity of Gram-negative bacteria. Cl and Cb intermediates showed lower toxicity than parent compounds. The findings help better understand the environmental impact of azole fungicides, their degradation, and toxicity. They also stress the need for reducing their uncontrolled release to the environment.

Nitrofurazone Removal from Water Enhanced by Coupling Photocatalysis and Biodegradation.

(1) Background: Environmental contamination with antibiotics is particularly serious because the usual methods used in wastewater treatment plants turn out to be insufficient or ineffective. An interesting idea is to support natural biodegradation processes with physicochemical methods as well as with bioaugmentation with efficient microbial degraders. Hence, the aim of our study is evaluation of the effectiveness of different methods of nitrofurazone (NFZ) degradation: photolysis and photodegradation in the presence of two photocatalysts, the commercial TiO2-P25 and a self-obtained Fe3O4@SiO2/TiO2 magnetic photocatalyst. (2) Methods: The chemical nature of the photocatalysis products was investigated using a spectrometric method, and then, they were subjected to biodegradation using the strain Achromobacter xylosoxidans NFZ2. Additionally, the effects of the photodegradation products on bacterial cell surface properties and membranes were studied. (3) Results: Photocatalysis with TiO2-P25 allowed reduction of NFZ by over 90%, demonstrating that this method is twice as effective as photolysis alone. Moreover, the bacterial strain used proved to be effective in the removal of NFZ, as well as its intermediates. (4) Conclusions: The results indicated that photocatalysis alone or coupled with biodegradation with the strain A. xylosoxidans NFZ2 leads to efficient degradation and almost complete mineralization of NFZ.

The metabolic pathways of polyhydroxyalkanoates and exopolysaccharides synthesized by Haloferax mediterranei in response to elevated salinity.

How polymer synthesis is mobilized or activated as a biological response of Haloferax mediterranei against hypertonic conditions remains largely unexplored. This study investigated the protein expression of H. mediterranei in response to high salinity by using isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomic analysis. The microbes were harvested at end of fermentation at the NaCl salinity of 75 and 250 g L-1. Among the identified 2123 proteins, 170 proteins were differentially expressed. Gene ontology annotation revealed that the highest number of proteins was annotated in biological process category, which was responsible for metabolic process, cellular component and catalytic activity. Differentially expressed proteins were belonged to the class of response to stimulus as well as catalytic activity and binding. Under high salinity conditions, three pathways were established as key responses of PHA and EPS production to hypertonic pressure. Two overexpressed proteins, beta-ketoacyl-ACP reductase and 3-hydroxyacyl-CoA dehydrogenase, enhanced the synthesis of PHAs. The serine-pyruvate transaminase and serine-glyoxylate transaminase were upregulated, thereby increasing the conversion of glucose to PHA. Downregulated levels of sulfate-adenylyl transferase and adenylyl-sulfate kinase could cause diminished EPS synthesis. This study could contribute to better understanding of the proteomic mechanisms of the synthesized polymers in defending against salt stress. SIGNIFICANCE: Haloferax mediterranei, a family member of halophilic archaea, is well known for its fermentative production of poly-ß-hydroxyalkanoates (PHAs). PHAs are natural polymers that exhibit great potential in a wide range of applications such as a good alternative to petroleum-based plastics and the biocompatible material. For decades, the functional role of PHAs synthesized by H. mediterranei is deemed to be carbon and energy reservations. The finding proved that differential production of PHA and EPS in H. mediterranei exposed to elevated salinity was caused by differential protein expression. This is the first report on how PHA and EPS synthesized by H. mediterranei is mobilized as the response of increased salinity, contributing to the understanding of halophilic archaea's response to hypertonic stress and the precise control of fermentation production. Despite its advantages as a PHA cell factory, H. mediterranei synthesized EPS simultaneously, thereby lowering the maximum yield of PHA production. Overall, salinity can be used as a vital microbial fermentation parameter to obtain the highest harvest of PHA, as well as the lowest EPS synthesis in industrial fermentation.

Investigation of the bacterial cell envelope nanomechanical properties after long-term exposure to nitrofurans.

Antibiotic residues in the environment may negatively affect biological communities in the natural ecosystems. However, their influence on environmental bacterial strains has not been thoroughly investigated. In this study, two representatives of 5-nitrofuran antibiotics (nitrofurantoin and furaltadone) were investigated in terms of their long-term influence on the cell envelopes of newly isolated environmental bacterial strains (Sphingobacterium caeni FTD2, Achromobacter xylosoxidans NFZ2 and Pseudomonas hibiscicola FZD2). A 12-month exposure of bacterial cells to nitrofurans at a concentration of 20 mg L-1 induced changes in the cell structure and texture (bacteria under stress conditions showed a loss of their original shape and seemed to be vastly inflated, the cells increased average surface roughness after exposure to NFT and FTD, respectively). AFM observations allowed the calculation of the bacterial cell nanomechanical properties. Significant increase in adhesion energy of bacteria after prolonged contact with nitrofurantoin was demonstrated. Changes in the permeability of bacterial membrane, fatty acids' composition and bacterial cell surface hydrophobicity were determined. Despite visible bacterial adaptation to nitrofurans, prolonged presence of pharmaceuticals in the environment has led to significant alterations in the cells' structures which was particularly visible in P. hibiscicola.

Surfactant addition in diesel oil degradation - how can it help the microbes?

PURPOSE: Despite wide research on bioremediation of hydrocarbon-contaminated soil, the mechanisms of surfactant-enhanced bioavailability of the contaminants are still unclear. The presented study was focused on the in-depth description of relationships between hydrocarbons, bacteria, and surfactants. In order to that, the biodegradation experiments and cell viability measurements were conducted, and the properties of cell surface were characterized. METHODS: MTT assay was employed to measure plant extracts toxicity to microbes. Then, membrane permeability changes were evaluated, followed by diesel oil biodegradation in the presence of surfactants measurements by GCxGC-TOFMS and PCR-RAPD analysis. RESULTS: Our study undoubtedly proves that different surfactants promote assimilation of different groups of hydrocarbons and modify cell surface properties in different ways. Increased biodegradation of diesel oil was observed when cultures with Acinetobacter calcoaceticus M1B were supplemented with Saponaria officinalis and Verbascum nigrum extracts. Interestingly, these surfactants exhibit different influences on cell surface properties and their viability in contrast to the other surfactants. Moreover, the preliminary analyses have shown changes in the genome caused by exposure to surfactants. CONCLUSIONS: The results indicated that the benefits of surfactant use may be related to deep modification at the omics level, not only that of cell surface properties and confirms the complexity of the interactions between bacterial cells, pollutants and surfactants.

Evaluating the Effect of Azole Antifungal Agents on the Stress Response and Nanomechanical Surface Properties of Ochrobactrum anthropi Aspcl2.2.

Azole antifungal molecules are broadly used as active ingredients in various products, such as pharmaceuticals and pesticides. This promotes their release into the natural environment. The detailed mechanism of their influence on the biotic components of natural ecosystems remains unexplored. Our research aimed to examine the response of Ochrobactrum anthropi AspCl2.2 to the presence of four azole antifungal agents (clotrimazole, fluconazole, climbazole, epoxiconazole). The experiments performed include analysis of the cell metabolic activity, cell membrane permeability, total glutathione level and activity of glutathione S-transferases. These studies allowed for the evaluation of the cells' oxidative stress response to the presence of azole antifungals. Moreover, changes in the nanomechanical surface properties, including adhesive and elastic features of the cells, were investigated using atomic force microscopy (AFM) and spectrophotometric methods. The results indicate that the azoles promote bacterial oxidative stress. The strongest differences were noted for the cells cultivated with fluconazole. The least toxic effect has been attributed to climbazole. AFM observations unraveled molecular details of bacterial cell texture, structure and surface nanomechanical properties. Antifungals promote the nanoscale modification of the bacterial cell wall. The results presented provided a significant insight into the strategies used by environmental bacterial cells to survive exposures to toxic azole antifungal agents.

Increased biological removal of 1-chloronaphthalene as a result of exposure: A study of bacterial adaptation strategies.

Effective biodegradation of hydrophobic pollutants, such as 1-chloronaphthalene, is strictly associated with the adaptation of environmental bacteria to their assimilation. This study explores the relation between the modifications of cell properties of bacteria belonging to Pseudomonas and Serratia genera resulting from a 12-month exposure to 1-chloronaphthalene, and their biodegradation efficiency. In the presented study, both bacterial strains exhibited higher (70%) degradation of this compound after exposure compared to unexposed (55%) systems. This adaptation can be associated with increased ratio of polysaccharides in the outer layers of bacterial cells, which was confirmed using infrared spectroscopy analysis. Additionally, the analysis of Raman spectra indicated conformational changes of extracellular carbohydrates from α- to ß-anomeric structure. Moreover, the changes in the cell surface hydrophobicity and cell membrane permeability differed between the strains and the Pseudomonas strain exhibited more significant modifications of these parameters. The results suggest that adaptation strategies of both tested strains are different and involve diverse reconstructions of the cell wall and membranes. The results provide a novel and deep insight into the interactions between environmental bacterial strains and chloroaromatic compounds, which opens new perspectives for applying spectrometric methods in investigation of cell adaptation strategies as a result of long-term contact with toxic pollutants.

Nitrofurantoin-Microbial Degradation and Interactions with Environmental Bacterial Strains.

The continuous exposure of living organisms and microorganisms to antibiotics that have increasingly been found in various environmental compartments may be perilous. One group of antibacterial agents that have an environmental impact that has been very scarcely studied is nitrofuran derivatives. Their representative is nitrofurantoin (NFT)-a synthetic, broad-spectrum antibiotic that is often overdosed. The main aims of the study were to: (a) isolate and characterize new microbial strains that are able to grow in the presence of NFT, (b) investigate the ability of isolates to decompose NFT, and (c) study the impact of NFT on microbial cell properties. As a result, five microbial species were isolated. A 24-h contact of bacteria with NFT provoked modifications in microbial cell properties. The greatest differences were observed in Sphingobacterium thalpophilum P3d, in which a decrease in both total and inner membrane permeability (from 86.7% to 48.3% and from 0.49 to 0.42 µM min-1) as well as an increase in cell surface hydrophobicity (from 28.3% to 39.7%) were observed. Nitrofurantoin removal by selected microbial cultures ranged from 50% to 90% in 28 days, depending on the bacterial strain. Although the isolates were able to decompose the pharmaceutical, its presence significantly affected the bacterial cells. Hence, the environmental impact of NFT should be investigated to a greater extent.

Environmental Aspects of the Use of Hedera helix Extract in Bioremediation Process.

This paper analyzes the impact of saponins from English ivy leaves on the properties of environmental bacterial strains and hydrocarbon degradation ability. For this purpose, two bacterial strains, Raoultella ornitinolytica M03 and Acinetobacter calcoaceticus M1B, have been used in toluene, 4-chlorotoluene, and α,α,α-trifluorotoluene biodegradation supported by Hedera helix extract. Moreover, theeffects of ivy exposition on cell properties and extract toxicity were investigated. The extract was found to cause minor differences in cell surface hydrophobicity, membrane permeability, and Zeta potential, although it adhered to the cell surface. Acinetobacter calcoaceticus M1B was more affected by the ivy extract; thus, the cells were more metabolically active and degraded saponins at greater amounts. Although the extract influenced positively the cells' viability in the presence of hydrocarbons, it could have been used by the bacteria as a carbon source, thus slowing down hydrocarbon degradation. These results show that the use of ivy saponins for hydrocarbon remediation is environmentally acceptable but should be carefully analyzed to assess the efficiency of the selected saponins-rich extract in combination with selected bacterial strains.

Biological impact of octyl d-glucopyranoside based surfactants.

Development of many branches of industry has stimulated the search for new, effective surfactants with interesting properties. Potential use of alkyl glucose derivatives on a large scale, raises questions about the possible risks associated with their entry into the natural environment. To be able to evaluate this risk, the aim of the study was to determine the physicochemical properties of octyl d-glucopyranoside and its three derivatives: N-(octyl d-glucopyranosiduronyl)aspartic acid, N-(octyl d-glucopyranosiduronyl)glicyne and octyl d-glucopyranosiduronic acid. Moreover, their biodegradability by pure bacterial strains and biocenosis present in river water was examined. While descriptions of sugar-based surfactants on microbial cells are limited, the essential element of the study was to determine the effect of surfactants on cell surface properties of microorganisms isolated from activated sludge and compare it to the effects of the petroleum based surfactants and the surfactants produced from renewable materials. The results obtained indicate that physicochemical properties of surface active agents differ depending on the presence of functional groups in the surfactants molecules. What is more, the presence of amino acid substituent in the derivatives of octyl d-glucopyranoside resulted in a slight decrease in the surfactants biodegradation efficiency, in comparison to the compounds that did not contain such a substituent, prolonging this process from 5 to 10 days. Interestingly, even relatively slightly different derivatives modified the cell surface properties in a different way. Importantly, the surfactants based on octyl d-glucopyranoside have less negative impact on environmental microorganism and better biodegradability than the surfactant synthesized from petroleum products.

Butylbenzene and tert-Butylbenzene-Sorption on Sand Particles and Biodegradation in the Presence of Plant Natural Surfactants.

The effects of hydrocarbons sorption on sand and saponins presence in the system on butylbenzene and tert-butylbenzene biological degradation was investigated. Additionally, the impact of saponins-containing plant extracts on environmental microorganisms was studied. Results of cell surface property measurements in samples with saponins only revealed changes in cell surface hydrophobicity, electrokinetic potential and membrane permeability when compared to corresponding values for glucose-grown microbes. Subsequently, in sorption experiments, the hydrocarbon adsorption kinetics in bacteria-free samples were better explained with the pseudo-second order kinetic model as compared to the pseudo-first order and intraparticular diffusion models. Moreover, the equilibrium data fitted better to the Freundlich isotherm for both benzene derivatives. In the samples combining hydrocarbons sorption and biological degradation in the presence of saponins, alkane-substituted hydrocarbons removal was accelerated from 40% to 90% after 14 days and the best surfactant in this aspect was S. officinalis extract.

Saponaria officinalis L. extract: Surface active properties and impact on environmental bacterial strains.

Plant-derived surfactants are characterised by low toxicity, high biodegradability and environmental compatibility. They therefore have many applications; for instance, they can be used in bioremediation to accelerate biodegradation processes, especially of hydrophobic pollutants. This paper analyses the properties of an extract from Saponaria officinalis L. containing saponins and its impact on bacterial strains isolated from soil, as well as its potential for application in hydrocarbon bioremediation. The tested extract from Saponaria officinalis L. contains gypsogenin, hederagenin, hydroxyhederagenin and quillaic acid aglycone structures and demonstrates good emulsification properties. Contact with the extract led to modification of bacterial cell surface properties. A decrease in cell surface hydrophobicity and an increase in membrane permeability were recorded in the experiments. An increase of up to 63% in diesel oil biodegradation was also recorded for Pseudomonas putida DA1 on addition of 1gL-1 of saponins from Saponaria officinalis L. Saponaria extract showed no toxic impact on the tested environmental bacterial strains at the concentration used in the biodegradation process.