Figure 2 - available via license: Creative Commons Attribution 4.0 International
Content may be subject to copyright.
![Diclofenac degradation by bacteria [9,10,55,56].](publication/368564936/figure/fig1/AS:11431281120584130@1676558667317/Diclofenac-degradation-by-bacteria-9-10-55-56.png)
Source publication
![Figure 2. Diclofenac degradation by bacteria [9,10,55,56].](publication/368564936/figure/fig1/AS:11431281120584130@1676558667317/Diclofenac-degradation-by-bacteria-9-10-55-56_Q320.jpg)
Diclofenac is one of the most popular non-steroidal anti-inflammatory drugs. Due to its over-the-counter availability and high consumption along with municipal and hospital wastewater, it enters the sewage treatment plant, where it is not completely degraded. This results in the appearance of diclofenac in the effluents from the treatment plant, an...
Context in source publication
Context 1
... described pathway leads through a series of oxidation reactions to homogentisic acid. The further oxidation of this acid through a quinone derivative leads to the end products: acetoacetic acid, fumaric acid and 4,6,7-trioxoоct-2-enedioic acid (Figure 2). The adaptation mechanisms of Rhodococcus ruber IEGM 346 to high concentrations of this drug are altered ζ potential of bacterial cells, increased cell hydrophobicity and total cell lipid content, formation of multicellular conglomerates, and altered surface-to-volume ratio [54]. ...
Citations
... The actual amount consumed, however, is difficult to determine due to their wide availability under different trade names (Panchal and Prince Sabina, 2023). Their therapeutic uses and benefits are numerous: the most common is the treatment of acute pain after surgery or chronic pain, but they are also used to treat osteoarthritis and rheumatoid arthritis (Wojcieszyńska et al., 2023). In addition, recently, they have been used to treat COVID-19 (Wojcieszyńska et al., 2022). ...
Drugs are chemical compounds used to treat and improve organic dysfunctions caused by diseases. These include analgesics, antibiotics, antidepressants, and antineoplastics. They can enter aquatic environments through wastewater streams, where their physico-chemical properties allow metabolites to distribute and accumulate. Current climate change and associated extreme weather events may significantly impact these substances' toxicity and aquatic organisms' sensitivity. Among the chemicals present in aquatic environments is the non-steroidal anti-inflammatory drug diclofenac (DIC), which the EU monitors due to its concentration levels. This study investigated the influence of temperature (control at 17 °C vs. 21 °C) on the effects of DIC (0 μg/L vs. 1 μg/L) in the mussel species Mytilus galloprovincialis. Significant results were observed between 17 and 21 °C. Organisms exposed to the higher temperature showed a decrease in several parameters, including metabolic capacity and detoxification, particularly with prolonged exposure. However, in some parameters, after 21 days, the M. galloprovincialis showed no differences from the control, indicating adaptation to the stress. The results of this study confirm that DIC concentrations in the environment, particularly when combined with increased temperatures, can produce oxidative stress and adversely affect M. galloprovincialis biochemical and physiological performance. This study also validates this species as a bioindicator for assessing environmental contamination with DIC. Beyond its direct impact on aquatic organisms, the presence of pharmaceuticals like DIC in the environment highlights the interconnectedness of human, animal, and ecosystem health, underscoring the One Health approach to understanding and mitigating environmental pollution.
... Ibu-2, Variovorax sp. Ibu-1) and (Wojcieszyńska et al., 2023). ...
Introduction
The increasing use of non-steroidal anti-inflammatory drugs (NSAIDs) has raised concerns regarding their environmental impact. To address this, understanding the effects of NSAIDs on bacteria is crucial for bioremediation efforts in pharmaceutical-contaminated environments. The primary challenge in breaking down persistent compounds lies not in the biochemical pathways but in capacity of bacteria to surmount stressors.
Methods
In this study, we examined the biodegradative activity, morphological and physiological changes, and ultrastructural adaptations of Rhodococcus cerastii strain IEGM 1243 when exposed to ibuprofen, diclofenac, and their mixture.
Results and Discussion
Our findings revealed that R. cerastii IEGM 1243 exhibited moderate biodegradative activity towards the tested NSAIDs. Cellular respiration assay showed higher metabolic activity in the presence of NSAIDs, indicating their influence on bacterial metabolism. Furthermore, catalase activity in R. cerastii IEGM 1243 exposed to NSAIDs showed an initial decrease followed by fluctuations, with the most significant changes observed in the presence of DCF and the NSAID mixture, likely influenced by bacterial growth phases, active NSAID degradation, and the formation of multicellular aggregates, suggesting potential intercellular synergy and task distribution within the bacterial community. Morphometric analysis demonstrated alterations in size, shape, and surface roughness of cells exposed to NSAIDs, with a decrease in surface area and volume, and an increase in surface area-to-volume ratio (SA/V). Moreover, for the first time, transmission electron microscopy confirmed the presence of lipid inclusions, polyphosphates, and intracellular membrane-like structures in the ibuprofen-treated cells.
Conclusion
These results provide valuable insights into the adaptive responses of R. cerastii IEGM 1243 to NSAIDs, shedding light on the possible interaction between bacteria and pharmaceutical compounds in the environment.
... Seven genes (ipfABDEFHI) were identified in the fosmid pFOS3G7 obtained from the chromosomal library of Sphingomonas sp. Ibu-2, which Schematic representation of DCF degradation by fungi or bacteria described in the literature (Bessa et al., 2017;Ivshina et al., 2019;Wojcieszyńska et al., 2023). Microbiology 11 frontiersin.org ...
The consumption of non-steroidal anti-inflammatory drugs (NSAIDs) have increased significantly in the last years (2020-2022), especially for patients in COVID-19 treatment. NSAIDs such as diclofenac, ibuprofen, and paracetamol are often available without restrictions, being employed without medical supervision for basic symptoms of inflammatory processes. Furthermore, these compounds are increasingly present in nature constituting complex mixtures discarded at domestic and hospital sewage/wastewater. Therefore, this review emphasizes the biodegradation of diclofenac, ibuprofen, and paracetamol by pure cultures or consortia of fungi and bacteria at in vitro, in situ, and ex situ processes. Considering the influence of different factors (inoculum dose, pH, temperature, co-factors, reaction time, and microbial isolation medium) relevant for the identification of highly efficient alternatives for pharmaceuticals decontamination, since biologically active micropollutants became a worldwide issue that should be carefully addressed. In addition, we present a quantitative bibliometric survey, which reinforces that the consumption of these drugs and consequently their impact on the environment goes beyond the epidemiological control of COVID-19.
... In recent years, the production and consumption of diclofenac has increased to an estimated level of 940 tons per year worldwide. It is administered in capsules, suppositories, tablets, intravenous solution and ointments (Wojcieszyńska et al., 2023). After administration, DCF is hepatically detoxified by hydroxylation and glucuronidation. ...
... Sixty-five percent of the oxidized metabolites are excreted by kidneys and the remainder is excreted in the bile as acyl glucuronide. A part of diclofenac is not metabolised, and, as a parent compound, reaches the sewage system at concentration levels of up to hundreds of μg/L (Wojcieszyńska et al., 2023). As reported by Sathishkumar and coauthors (2020), conventional sewage treatment plants, with limited physico-chemical removing methods, are not able to efficiently eliminate DCF residues which are unavoidably found in freshwater systems. ...
This study assessed the eco-genotoxic impact of diclofenac (DCF) in sentinel species of the freshwater ecosystem. DCF residues are found in freshwater from few ng/L to tens of μg/L due to the inability of conventional wastewater treatment plants to ensure removal efficiency of the drug. An ample body of literature reports on the acute toxicity of DCF in non-target organisms without addressing potential chronic long-term effects on organisms at actual, environmental concentrations. Herein, assessment for acute and chronic toxicity was performed on organisms in vivo exposed to DCF, specifically on the green alga Raphidocelis subcapitata, the rotifer Brachionus calyciflorus and the crustacean Ceriodaphnia dubia. Furthermore, potential DNA damage and expression of antioxidant genes (MnSOD, Cu/ZnSOD and CAT) were evaluated in crustacean neonates. The toxicological risk of DCF was assessed as well as its. GENOTOXIC RISK: The acute toxicity was observed at concentrations far from those of environmental concern. Rotifers and crustaceans were much more chronically sensitive than the algae to DCF, observing besides, the median effect concentrations at tens of μg/L. In crustaceans, DNA damage was noted at units of μg/L, revealing concentrations of environmental concern. The dysregulated activity of SOD and CAT also showed the ability of DCF to provoke oxidative stress. On assessment of environmental risk, the chronic Risk Quotient (RQ) was above the threshold value of 1. Nevertheless, the genotoxic RQ was significantly greater than the chronic RQ, thus, the need of regulatory bodies to acknowledge the genotoxic impact as an environmental risk factor. To our knowledge, this study is the first investigation to perform environmental genotoxic risk assessment of DCF.
... This contributes to their high consumption and, consequently, their presence in sewage and the environment. Among others, naproxen and diclofenac have been found in European waters at concentration ranges of 3-753 ng/L and 1-429 ng/L, respectively [1][2][3][4]. Concentrations observed in the environment are not the root of acute toxicity. However, it has been clearly demonstrated that they lead to chronic toxicity. ...
... On the other hand, diclofenac has two aromatic rings linked together by an amine bridge. Although the literature indicates a relatively easy cleavage of this compound into two rings, the deactivating effect of chlorine substituents in diclofenac significantly slows down the degradation process [1,2,4]. ...
... Phenol is a good carbon source for the B1 strain [17]. However, immobilization significantly affects the growth kinetics of the strains due to the limited availability of the substrate for cells [4,15]. The lack of changes in the rate of ibuprofen degradation in systems with different phenol concentrations may be related to the fact that its degradation proceeds through hydroxylation with the participation of aliphatic monooxygenase and transformation to 1,4-hydroquinone, for which acyl-CoA synthase is responsible, the enzymes of which are not induced by phenol [34,38]. ...
Among the micropollutants identified in the environment, non-steroidal anti-inflammatory drugs (NSAIDs) dominate more and more often. This is due to both the high consumption and low efficiency of biological wastewater treatment plants, where the initial transformation of NSAIDs most often takes place. The solution to the problem may be using preparations supporting activated sludge in sewage treatment plants in the biodegradation of NSAIDs. Therefore, the research aimed to develop a biopreparation stimulating the activated sludge of the sewage treatment plant to decompose paracetamol and selected NSAIDs. This biopreparation is based on strains of Stenotrophomonas maltophilia KB2, Planococcus sp. S5, Bacillus thuringiensis B1(2015b), and Pseudomonas moorei KB4 immobilized on a plant sponge. As a result of the tests, it was shown that the optimal species composition of the proposed preparation includes all tested strains immobilized on a carrier with a mass of 1.2 g/L. The system optimization showed that the optimal amount of strains on the carrier was 17 mg/g of the carrier, 15 mg/g of the carrier, 18 mg/g of the carrier, and 20 mg/g of the carrier for KB4, B1(2015b), KB2, and S5, respectively. The presence of phenol stimulated the degradation of the tested drugs, and this effect deepened with increasing phenol concentration. At the same time, the degradation rate of the mixture of NSAIDs in the presence of phenol did not depend on the amount of biomass. The lack of inhibition in the presence of an additional co-contaminant, i.e., phenol, indicates that the preparation constructed in this way has a chance of being used in sewage treatment plant systems, where introduced strains are exposed to various aromatic compounds.
... In the literature, different genera of both Gram-positive and Gram-negative bacteria were identified as potential DCF-degrading bacteria, such as Klebsiella, Raoultella, Bacillus, and Rhodococcus [20,28,30]. Despite the diversity of the identified bacterial strains, few bacterial isolates were able to completely degrade DCF due to the difficulty of degradable intermediates generated during biodegradation [31]. In this study, two strains of the four selected bacteria were new for DCF biodegradation, namely, A. spanius strain S11 and A. piechaudii S18. ...
... The results obtained from the identification of intermediate metabolites by GC-MS during DCF biodegradation suggested that the bacterial degradation reactions that occurred were mostly multidirectional [31]. The primary hydroxylation of DCF is considered a bottleneck step in DCF metabolites, and the detection of 4 -OH-DCF and 5 -OH-DCF in all investigated isolates indicated that CYP450 monooxygenase is a key enzyme in this degradation pathway [14,30]. ...
... In fact, most of the described microorganisms capable of DCF biodegradation transformed it into a more toxic hydroxylated form. However, only a few isolates could further cleave the aromatic structure of this hydroxylated DCF [2,31]. The formation of quinone imine derivatives of 5-hydroxy diclofenac (DF-2,5-QI) in the A. spanius S11 culture via the action of laccase was previously demonstrated [34]. ...
The accumulation of xenobiotic compounds in different environments interrupts the natural ecosystem and induces high toxicity in non-target organisms. Diclofenac is one of the commonly used pharmaceutical drugs that persist in the environment due to its low natural degradation rate and high toxicity. Therefore, this study aimed to isolate potential diclofenac-degrading bacteria, detect the intermediate metabolites formed, and determine the enzyme involved in the degradation process. Four bacterial isolates were selected based on their ability to utilize a high concentration of diclofenac (40 mg/L) as the sole carbon source. The growth conditions for diclofenac degradation were optimized, and bacteria were identified as Pseudomonas aeruginosa (S1), Alcaligenes aquatilis (S2), Achromobacter spanius (S11), and Achromobacter piechaudii (S18). The highest percentage of degradation was recorded (97.79 ± 0.84) after six days of incubation for A. spanius S11, as analyzed by HPLC. To detect and identify biodegradation metabolites, the GC-MS technique was conducted for the most efficient bacterial strains. In all tested isolates, the initial hydroxylation of diclofenac was detected. The cleavage step of the NH bridge between the aromatic rings and the subsequent cleavage of the ring adjacent to or in between the two hydroxyl groups of polyhydroxylated derivatives might be a key step that enables the complete biodegradation of diclofenac by A. piechaudii S18, as well as P. aeruginosa S1. Additionally, the laccase, peroxidase, and dioxygenase enzyme activities of the two Achromobacter strains, as well as P. aeruginosa S1, were tested in the presence and absence of diclofenac. The obtained results from this work are expected to be a useful reference for the development of effective detoxification bioprocesses utilizing bacterial cells as biocatalysts. The complete removal of pharmaceuticals from polluted water will stimulate water reuse, meeting the growing worldwide demand for clean and safe freshwater.
... The conversion rate remained at 100% after repeating the operation for 41 batches of sPBR. A review of diclofenac biodegradation by microorganisms and immobilized systems was presented [12]. It showed that immobilized fungal and bacterial systems can achieve complete degradation of diclofenac by a metabolic relay that avoids the accumulation of toxic intermediates. ...
Biocatalysis, which can be performed by whole cells and isolated enzymes, has become a topic of public interest for its potential use in the chemical industry in manufacturing, monitoring, and waste management. Enzymes are proteins that organisms produce to catalyze the biochemical reactions needed for life. However, the isolation and purification of enzymes may be costly and time-consuming, and cofactors may need to be added or recovered. An alternative approach is to use whole cells as “Microbial Biocatalysts” to perform multiple enzyme reactions in a single strain and regenerate cofactors internally. Whole-cell biocatalysts can be used for different types of processes, such as biotransformation and fermentation. They involve one or more steps of biocatalysis to produce valuable chemicals through biosynthesis/biotransformation or degrade organic pollutants completely
Significant concentrations of emerging xenobiotics, like diclofenac (DCF), possessing severe irreversible eco-
toxicological threats, has been detected in aquatic systems worldwide, raising the concerns. This present investi-
gation is intended to explore an efficient solution to support the existing wastewater treatment policies to handle
DCF contamination by bacteria-mediated biotransformation. DCF-tolerant bacterial strains were isolated from
pharmaceutical wastewater and selected based on their non-virulence nature and degradation ability. Among
those, Pseudomonas sp. DCα4 was found to be the most dominant DCF degrader exhibiting 99.82% removal of
DCF confirmed by HPLC after optimization of temperature at 30.02 °C, pH at 6.9, inoculum of 4.94%, and time
68.02 h. The degradation kinetics exhibited the process of DCF degradation followed a first-order kinetics with k
of 0.108/h and specific degradation rate of 0.013/h. Moreover, the enzyme activity study indicated predominant
hydrolase activity in the DCF treatment broth of DCα4, implying hydrolysis as the main force behind DCF bio-
transformation. HRMS analysis confirmed the presence of 2-hydroxyphenylacetic acid, 1,3-dichloro,2-amino, 5-
hydroxybenzene, and benzylacetic acid as major intermediates of DCF biodegradation indicating non-specific hy-
drolysis of DCF. Whole genome analysis of most related strains which were confirmed by near full 16S rRNA gene
sequence homology study, predicted involvement of different N–C bond hydrolase producing genes like puud,
atzF, astB, nit1, and nylB. The ecotoxicological study using Aliivibrio fischeri exhibited 47.51% bioluminescence
inhibition by DCF-containing broth which was comparable to the same caused by 1 mg/mL of K2Cr2O7 whereas
remediated broth exhibited only 0.51% inhibition implying reduction of the ecotoxic load caused by DCF conta-
mination. Cost analysis revealed that possible integration of the process with existing ones would increase per
litre expense by $0.45. These results indicated that the described process of DCF biodegradation using the super-
degrader DCα4 would be an advancement of existing pharmaceutical wastewater treatment processes for DCF
bioremediation.
