Effect of phenol formaldehyde-associated microplastics on soil microbial community, assembly, and functioning.

Increasing investigations explore the effects of plastic pollutants on bacterial communities, diversity, and functioning in various ecosystems. However, the impact of microplastics (MPs) on the eukaryotic community, microbial assemblages, and interactions is still limited. Here, we investigated bacterial and micro-eukaryotic communities and functioning in soils with different concentrations of phenol formaldehyde-associated MPs (PF-MPs), and revealed the factors, such as soil properties, microbial community assembly, and interactions between microbes, influencing them. Our results showed that a high concentration (1%) of PF-MPs decreased the microbial interactions and the contribution of deterministic processes to the community assembly of microbes, and consequently changed the communities of bacteria, but not eukaryotes. A significant and negative relationship was determined between N2O emission rate and functional genes related to nitrification, indicating that the competitive interactions between functional microbes would affect the nitrogen cycling of soil ecosystem. We further found that vegetable biomass weakly decreased in treatments with a higher concentration of PF-MPs and positively related to the diversity of micro-eukaryotic communities and functional diversity of bacterial communities. These results suggest that a high concentration of the PF-MPs would influence crop growth by changing microbial communities, interactions, and eukaryotic and functional diversity. Our findings provide important evidence for agriculture management of phenol formaldehyde and suggest that we must consider their threats to microbial community compositions, diversity, and assemblage in soils due to the accumulation of PF-MPs widely used in the field.

Impact of sublethal phenol in freshwater fish Labeo rohita on biochemical and haematological parameters.

Phenol, an aromatic chemical commonly found in domestic and industrial effluents, upon its introduction into aquatic ecosystems adversely affects the indigenous biota, the invertebrates and the vertebrates. With the increased demand for agrochemicals, a large amount of phenol is released directly into the environment as a byproduct. Phenol and its derivatives tend to persist in the environment for longer periods which in turn poses a threat to both humans and the aquatic ecosystem. In our current study, the response of Labeo rohita to sublethal concentrations of phenol was observed and the results did show a regular decrease in biochemical constituents of the targeted organs. Exposure of Labeo rohita to sublethal concentration of phenol (22.32 mg/L) for an epoch of 7, 21 and 28 days shows a decline in lipid, protein, carbohydrate content and phosphatase activity in target organs such as the gills, muscle, intestine, liver and kidney of the fish. The present study also aims to investigate the toxic effects of phenol with special reference to the haematological parameters of Labeo rohita. At the end of the exposure period, the blood of the fish was collected by cutting the caudal peduncle with a surgical scalpel. And it was observed that the red blood corpuscle count (RBC), white blood corpuscle (WBC), haemoglobin count (Hb), packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC) values showed a decline after exposure to phenol for 7 days, while white blood corpuscle (WBC) shows an increased count. At 21 days and 28 days, all the haematological parameters showed a significant decrease.

Valorization of phenol contaminated wastewater for lipid production by Rhodosporidium toruloides 9564T.

Phenol is one of the most common hazardous organic compound presents in several industrial effluents which directly affects the aquatic environment. The present study envisaged the phenol biodegradation and simultaneous lipid production along with its underlying mechanism by oleaginous yeast Rhodosporidium toruloides 9564T. Experiments were designed using simulated wastewater by varying phenol concentration in the range of 0.25-1.5 g/L and inoculum size of 1, 5, and 10% with and without glucose. The oleaginous yeast was found to completely degrade up to 0.75 g/L phenol with lipid accumulation of 26.3%. Phenol at > 0.5 g/L severely inhibited the growth of R. toruloides 9564T at 1% and 5% inoculum size. Phenol toxicity up to 0.75 g/L can be overcome by increasing inoculum size to 10%. The maximum specific growth rate (µmax) and phenol degradation rate (qmax) were found to be 0.0717 h-1 and 0.01523 h-1, respectively. The enzymatic pathway study suggested that R. toruloides 9564T follows an ortho cleavage pathway for phenol degradation and lipid accumulation. Phytotoxicty and cytotoxicity tests for treated and untreated samples clearly demonstrated a decline in toxicity of the treated wastewater. R. toruloides brought about an important paradigm shift toward a circular economy in which industrial wastewater is considered a valuable resource for bioenergy production.

Synergistic Interaction of Polycyclic Aromatic Hydrocarbons, Phthalate Esters, or Phenol on DNA Adduct Formation by Aristolochic Acid I: Insights into the Etiology of Balkan Endemic Nephropathy.

Balkan endemic nephropathy (BEN) is a multifactorial environmental disease, with chronic exposure to aristolochic acids (AAs) through AA-contaminated food being one of the major etiological mechanisms. However, the bulk of previous research has only focused on investigating the possible roles of individual pollutants in disease development and the etiological mechanism of BEN remains controversial. In this study, we investigated the exposure concentration and duration dependence of coexposure to phthalate esters and lignite coal-derived phenol and polycyclic aromatic hydrocarbons (PAHs) on the metabolism and DNA adduct formation of aristolochic acid I (AAI). Results showed that both the metabolic activation and DNA adduct formation of AAI in cultured human kidney cells were affected by their coexposure to the above-mentioned environmental pollutants. Furthermore, our results suggest that chemicals leached from lignite coal likely played a role by triggering AA-activating enzymes to produce more of the promutagenic DNA adducts, thus further elevating the nephrotoxicity and carcinogenicity of AAs and increasing the risk of BEN. It is believed that the results of this study provide a better understanding of the etiological mechanism of BEN and offer insights into methods and policies to lower the risk of this devastating disease.

Removal of ammonia and phenol from saline chemical wastewater by ionizing radiation: Performance, mechanism and toxicity.

Saline chemical wastewater containing ammonia and toxic organic pollutants has been a challenge for conventional wastewater treatment technology. Advanced treatment is thus required. In this study, the removal of ammonia and phenol in saline chemical wastewater by radiation was investigated in detail. The results showed that chloridion in saline chemical wastewater could be transferred to •Cl and •ClO by radiation, which promoted ammonia oxidation, but inhibited phenol degradation. Solution pH affected the types of reactive species, which further affected the removal of ammonia and phenol. When ammonia and phenol co-existed in saline chemical wastewater, the removal efficiency of ammonia was depressed compared to that in the absence of phenol. Similarly, the phenol removal efficiency was also depressed in the presence of ammonia when the solution pH was lower than 7.0. Interestingly, the phenol removal efficiency was improved with increase of either chloridion concentration (2-8 g/L) or dose (2-5 kGy), which was attributed to the formation of intermediate nitrogen-centered radicals that can react with phenol. In addition, the intermediate products of phenol degradation under different conditions were identified. The acute toxicity of saline chemical wastewater after radiation treatment was evaluated. The results of this study could provide an insight into the removal of ammonia and phenol from saline chemical wastewater by radiation technology.

Adaptation of phenol-degrading Pseudomonas putida KB3 to suboptimal growth condition: A focus on degradative rate, membrane properties and expression of xylE and cfaB genes.

Detailed characterization of new Pseudomonas strains that degrade toxic pollutants is required and utterly necessary before their potential use in environmental microbiology and biotechnology applications. Therefore, phenol degradation by Pseudomonas putida KB3 under suboptimal temperatures, pH, and salinity was examined in this study. Parallelly, adaptive mechanisms of bacteria to stressful growth conditions concerning changes in cell membrane properties during phenol exposure as well as the expression level of genes encoding catechol 2,3-dioxygenase (xylE) and cyclopropane fatty acid synthase (cfaB) were determined. It was found that high salinity and the low temperature had the most significant effect on the growth of bacteria and the rate of phenol utilization. Degradation of phenol (300 mg L-1) proceeded 12-fold and seven-fold longer at 10 °C and 5% NaCl compared to the optimal conditions. The ability of bacteria to degrade phenol was coupled with a relatively high activity of catechol 2,3-dioxygenase. The only factor that inhibited enzyme activity by approximately 80% compared to the control sample was salinity. Fatty acid methyl ester (FAMEs) profiling, membrane permeability measurements, and hydrophobicity tests indicated severe alterations in bacteria membrane properties during phenol degradation in suboptimal growth conditions. The highest values of pH, salinity, and temperature led to a decrease in membrane permeability. FAME analysis showed fatty acid saturation indices and cyclopropane fatty acid participation at high temperature and salinity. Genetic data showed that suboptimal growth conditions primarily resulted in down-regulation of xylE and cfaB gene expression.

Sublethal effects of phenol on histology of selected organs of freshwater fish Mystus vittatus.

Acute toxicity (96 h LC50) of phenol was analyzed in the cat fish Mystus vittatus in static bio-assay over a 96-h exposure period using probit method. The 24, 48, 72, and 96 h LC50 values (with 95% confidence limits) of phenol for fingerling catfish were found out as 13.98, 13.17, 12.62, and 12.21 mg/l respectively. Investigations pertaining to the histopathological sections have shown high degree of pathological lesions observed in various parts like gill, liver intestine, and kidney of the fish species. Analysis of gill section revealed observable changes in the experimental species such as fusion, malformation at the tip of secondary lamellae, vacuolation, hyperplasia, and epithelial damage. Exposure of phenol showed cytoplasmic vacuolation, tissue damage, and loss of hepatic cell wall in the liver of experimental organism. Lesions of tissue damage at the epithelial site, inflammation, and clumping of adjacent villi made of columnar epithelium have been observed in the intestine of fish, and also the excretory part of the fish kidney revealed various changes like glomerular atrophy, damage of Bowman's capsule, vacuolization, and degeneration of renal epithelium. The current study on histological changes observed in the experimental organisms has thrown light on the current scenario which poses threat and danger to the whole aquatic ecosystem, and this study plays a vital role in assessing the aquatic pollution.

Application of two bioassays as potential indicators of phenol phytoremediation efficiency by tobacco hairy roots.

Phenol is one of the contaminants most frequently found in the environment and it is considered a priority pollutant due to their toxic effects. Hairy roots (HR) constitute a good model tool for the removal of this contaminant. In this work, phenol removal using wild type (WT) and double transgenic (DT) Nicotiana tabacum HR was performed with high efficiency (60-80%, for 25-250 mg L-1 phenol solutions, respectively). After phytoremediation process, the toxicity of post removal solutions (PRS) was evaluated through two-toxicity test belonging to two trophic levels, Lactuca sativa test and Rhinella arenarum (AMPHITOX). Toxicity of PRS showed variable results since these solutions were less toxic to L. sativa seeds compared to R. arenarum embryos, which could be attributed to different sensitivities of the exposed organisms. Although PRS obtained using WT and DT HR reduced phenol phytotoxicity on L. sativa seeds, WT PRS were even less toxic than DT PRS according to this test. Regarding AMPHITOX, HR culture medium without phenol but incubated with HR and phenol PRS exerted a toxic effect on the embryos, which could be related to the presence of toxic products derived from HR metabolism. The results demonstrated that an efficient phenol removal is not always accompanied by a considerable reduction of the solution toxicity and therefore, the use of organisms from different trophic levels to evaluate the toxicity after the removal process gains importance.

Effect of Fe3O4 nanoparticles exposure on the treatment efficiency of phenol wastewater and community shifts in SBR system.

Increasing magnetic Fe3O4 nanoparticles (Fe3O4 NPs) application has aroused concern about its potential environmental toxicity. During acute and chronic exposure, key enzymes involved in phenol biodegradation were promoted at 0-600 mg/L Fe3O4 NPs, while were inhibited at 800 mg/L Fe3O4 NPs, correspondingly affected phenol degradation efficiency. Lactic dehydrogenase (LDH) increased when Fe3O4 NPs exceeded 600 mg/L, indicated the more severe cell rupture at high Fe3O4 NPs concentration. At the same Fe3O4 NPs concentration, the removal of EPS further inhibited key enzymes, decreased phenol degradation, and increased LDH, indicating that the existence of EPS relieved the adverse effects on microorganisms. Spectroscopic analysis showed that protein and polysaccharide associated bonds in EPS decreased at 0-600 mg/L Fe3O4 NPs, while increased when Fe3O4 NPs exceeded 600 mg/L, which was in accordance with EPS content. Biopolymer-degrading and phenol-degrading genera increased at 0-600 mg/L Fe3O4 NPs, while decreased at Fe3O4 NPs exceeded 600 mg/L, which conformed to EPS content and phenol degradation efficiency.

Preparation and characterization of rice husk adsorbents for phenol removal from aqueous systems.

Rice husk is a base adsorbent for pollutant removal. It is a cost-effective material and a renewable resource. This study provides the physicochemical characterization of chemically and thermally treated rice husk adsorbents for phenol removal from aqueous solutions. We revealed new functional groups on rice husk adsorbents by Fourier transform infrared spectroscopy, and observed major changes in the pore structure (from macro-mesopores to micro-mesopores) of the developed rice husk adsorbents using scanning electron microscopy. Additionally, we studied their surface area and pore size distribution, and found a greater enhancement of the morphological structure of the thermally treated rice husk compared with that chemically treated. Thermally treated adsorbents presented a higher surface area (24-201 m2.g-1) than those chemically treated (3.2 m2.g-1). The thermal and chemical modifications of rice husk resulted in phenol removal efficiencies of 36%-64% and 28%, respectively. Thus, we recommend using thermally treated rice husk as a promising adsorbent for phenol removal from aqueous solutions.

Verification on the Developmental Toxicity of Short-term Exposure to Phenol in Rats.

OBJECTIVE: To verify the health advisory for short-term exposure to phenol. METHODS: The method of this validation experiment was the same as the US Environmental Protection Agency (EPA) methodology for toxicology experiments used to determine phenol drinking water equivalent level (DWEL). Pregnant female Sprague-Dawley rats were administered phenol in distilled water by gavage at daily doses of 15, 30, 60, 120, and 240 mg/kg body weight (b.w.) from implantation (the 6th day post-mating) to the day prior to the scheduled caesarean section (the 20th day of pregnancy). The following information was recorded: general behavior; body weight; number of corpus luteum, live birth, fetus, stillbirth, and implantation; fetal gender; body weight; body length; tail length; and abnormalities and pathomorphological changes in the dams. RESULTS: In the 60 mg/kg b.w. dose group, the mortality of pregnant rats increased with increasing doses, suggesting maternal toxicity. Fetal and placental weights decreased as phenol dose increased from 30 mg/kg b.w., and were significantly different compared those in the vehicle control group, which suggested developmental toxicity in the fetuses. However, the phenol-exposed groups showed no significant change in other parameters compared with the vehicle control group ( P > 0.05). CONCLUSION: Despite using the same method as the US EPA, a different NOEAL of 15 mg/(kg·d) was obtained in this study.

Deriving hazardous concentrations of phenol in soil ecosystems using a species sensitivity distribution approach.

Phenol is widely used in many industries, and chemical accidents involving phenol have frequently occurred around the world, resulting in the investigation of phenol toxicity in humans, mammals, and aquatic organisms. However, very few studies have investigated phenol toxicity in terrestrial ecosystems. Therefore, we investigated the acute and chronic toxicity of phenol using various soil organisms, including Chlamydomonas reinhardtii, Chlorococcum infusionum, Folsomia candida, Oryza sativa, Raphanus sativus, Pinus densiflora, and Eisenia fetida. The data obtained were used to calculate hazardous concentrations for 5% of species (HC5) for phenol based on a species sensitivity distribution approach. The acute and chronic soil HC5 values for phenol were estimated to be 18.4 and 0.3 mg kg-1, respectively. To the best of our knowledge, this is the first study to conduct battery testing and calculate hazardous concentrations to assess the risk posed by phenol in terrestrial ecosystems. The results can be used to establish standards or strategies to protect terrestrial environments against unintended phenol contamination.

Formation of new PHE plasmids in pseudomonads in a phenol-polluted environment.

Several years ago, a laboratory-constructed plasmid with a single-component phenol monooxygenase gene (pheBA operon) flanked by two IS elements was released to a phenol-polluted area. During the following years, we found in the test area widely distributed pheBA operon-containing bacteria. The new pheBA+ strains belong predominantly to the Pseudomonas fluorescens group, and they did not arise via selection of the released PHE plasmid. On the contrary, the formation of several different types of PHE plasmids occurred, namely pPHE101 (60,958 bp) from the IncP-9 group, non-transferable plasmid pPHE69 (44,717 bp), mobilizable plasmid pPHE20 (39,609 bp) and the IncP-7 type plasmid pPHE24ΔpheBA (120,754 bp), in which the pheBA operon was translocated from the plasmid to the chromosome. In two cases, PHE plasmid-bearing strains exist in a multi-plasmid state, also containing the non-catabolic plasmids pG20 (133,709 bp) and pG69 (144,433 bp) with backbones sharing 97% DNA identity and with redundant genes for the initiation of replication, repA1and repA2, of which only one was active. Seemingly, several other plasmids and bacterial features besides the pheBA operon were involved in selective distribution of catabolic operons in the natural environment. The comparison of the genetic structure of plasmids and IS elements' functions, as well as resistance to heavy metals of seven completely sequenced plasmids, are discussed.

Impact of phenol on the glycerophospholipid turnover and potential role of circadian clock in the plant response against this pollutant in tobacco hairy roots.

Glycerophospholipids (GPLs) from cell membranes (CM) are a proper source for the synthesis of lipid messengers able to activate signal pathways that will define the plant survival under changing and stressful environmental conditions. Little is known about how GPLs metabolism (GPLsM) is regulated and the effects of phenol treatment on GPLs composition. In this work, we studied the effects of phenol both on GPLs turnover and on the expression of GPLsM-related genes potentially regulated by the circadian clock, using tobacco hairy root cultures (HRC). Phenol decreased the total PC levels and increased PE, PG and CL levels in the dark phase. Different molecular species of PC and PE showed the same trend than the total PC and PE upon phenol treatment. Besides, significant differences in the expression of all studied genes related to GPLsM were found. NtCCT2 expression was affected at all analyzed times while NtPECT1 and NtAAPT1 showed similar expression patterns. NtCDS1, NtPGPS2 and NtCLS genes showed significant and differential expression profiles both in untreated and treated HRC. PECT1 and NtPGPS2 genes seem to conserve a circadian expression profile mainly in untreated HRC. However, phenol was able to modify the GPLs composition and the expression of genes related to GPLs synthesis. The GPLs modification could be explained by the up-regulation of NtPECT1, NtAAPT1 and NtCLS genes during the dark phase, suggesting for being a crucial moment for HRC to trigger an adaptive response against this organic pollutant.

Growth behavior, glucose consumption and phenol removal efficiency of Chlorella vulgaris under the synergistic effects of glucose and phenol.

The use of algae is an effective approach to remove phenol and its derivatives from polluted water. The growth behavior, glucose consumption and phenol removal efficiency of Chlorella vulgaris under the synergistic effects of glucose and phenol were investigated. The evolutions of tolerance and removal efficiency of C. vulgaris to phenol under different trophic modes and glucose contents were observed. The results revealed that growth of C. vulgaris were inhibited with the increase of phenol from 0 to 400 mg L-1 in culture media; the tolerance to phenol enhanced with the addition of glucose from 2 to 10 g L-1, while glucose consumption was inhibited with the increase of phenol content; phenol removal efficiency varied with glucose concentrations in mixotrophic media. The finding suggested that phenol inhibited the growth of C. vulgaris and glucose assimilation under mixotrophic cultivation, while appropriate glucose addition could enhance the tolerance of C. vulgaris to phenol and affect the phenol removal efficiency.

A quantitative evaluation method for wastewater toxicity based on a microbial fuel cell.

A microbial fuel cell (MFC) was successfully developed as a toxicity biomonitoring system to enable a quick response to wastewater with unknown toxicity in toxic events. The objective was to quantitatively assess the toxicity of wastewater by a rapid method. Different concentrations of formaldehyde were introduced into the anode chamber, which led to different stages of voltage change. A relationship between the linear slope of the voltage drop stage and the formaldehyde concentration was established through dose-response fitting results. This relationship makes it possible to convert an unknown toxicity of wastewater into the equivalent concentration of formaldehyde. The minimum detection limit in this study was 13 mg/L formaldehyde equivalents. As the toxicity of the wastewater increased, the test time could be reduced to as low as 921 s or even shorter, with a detection error of 3-12 mg/L. By using this evaluation method, oxidized tail gas scrubber wastewater was identified as the main toxic wastewater component in a phenol acetone production plant.

Effects of phenol on glutathione S-transferase expression and enzyme activity in Chironomus kiiensis larvae.

Detoxifying enzyme mRNAs are potentially useful stress biomarkers. Glutathione S-transferase (GST) metabolises lipophilic organic contaminants and mitigates oxidative damage caused by environmental pollutants. Herein, 12 Chironomus kiiensis GSTs (CkGSTs1-6, CkGSTt1-2, CkGSTd1-2, CkGSTm1-2) were cloned and grouped into sigma, theta, delta and microsomal subclasses. Open reading frames (450-699 bp) encode 170-232 amino acid proteins with predicted molecular masses of 17.31-26.84 kDa and isoelectric points from 4.94 to 9.58. All 12 GSTs were expressed during all tested developmental stages, and 11 displayed higher expression in fourth-instar larvae than eggs. GST activity after 24 h of phenol exposure was used to estimate environmental phenol contamination. After exposure to sublethal concentrations of phenol for 48 h, expression and activity of CkGSTs were inhibited in C. kiiensis larvae. Expression of CkGSTd1-2 and CkGSTs1-2 varied with phenol concentration, indicating potential use as biomarkers for monitoring environmental phenol contamination.

Involvement of brain GABAAR-coupled Cl-/HCO3--ATPase in phenol-induced the head-twitching and tremor responses in rats.

Phenol-induced neurotoxicity manifests as twitching/tremor and convulsions, but its molecular mechanisms underlying the behavioral responses remain unclear. We assessed the role of the brain Cl-/HCO3--ATPase in behavioral responses in rats following an in vivo intraperitoneal injection of phenol (20-160 mg/kg). Low concentrations of phenol (20-80 mg/kg) increased the ATPase activity as well as the head twitching responses in rat, whereas higher phenol concentrations (>60 mg/kg) increased the tremor but reduced the ATPase activity. At phenol concentrations >120 mg/kg, no ATPase activity was detected. Phenobarbital (10 mg/kg) and picrotoxin (1 mg/kg) as well as o-vanadate (2 mg/kg), significantly prevented (˜55-70%) the phenol-induced change in the behavioral responses and completely restored the enzyme activity. In vitro experiments confirmed that phenol stimulated the Cl-/HCO3--ATPase activity at low concentrations, but had no stimulating effect on other transport ATPases. Low doses of phenol increased the formation of phosphoprotein and the rate of ATP-consuming Cl- transport by the reconstituted enzyme. The present findings provide evidence that phenol-induced neurotoxicity involves the Cl-/HCO3--ATPase in the behavioral responses in mammals and indicate the potential benefit of this enzyme as a target for the treatment of head twitching and other types of tremor diseases.

Identification of RNA biomarkers for chemical safety screening in neural cells derived from mouse embryonic stem cells using RNA deep sequencing analysis.

Chemical safety screening requires the development of more efficient assays that do not involve testing in animals. In vitro cell-based assays are among the most appropriate alternatives to animal testing for screening of chemical toxicity. Most studies performed to date made use of mRNAs as biomarkers. Recent studies have however indicated the presence of many unannotated non-coding RNAs (ncRNAs) in the transcriptome that do appear to encode proteins. In the present study, we performed whole-transcriptome sequencing analysis (RNA-Seq) to identify novel RNA biomarkers, including ncRNAs, which showed marked responses to the toxicity of nine chemicals. Chemical safety screening was performed in cell-based assays using mouse embryonic stem cell (mESC)-derived neural cells. Marked responses in the expression of some ncRNAs to the chemical compounds were observed. The results of the present study suggested that ncRNAs may be useful in chemical safety screening as novel RNA biomarkers.