Biodegradation Of Antibiotics Research Articles

Assessing Antibiotics Biodegradation and Effects at Sub-inhibitory Concentrations by Quantitative Microbial Community Deconvolution

Antibiotics in the environment cause widespread concern as a result of their potent inhibitory action on microbial growth and their role in potentially creating selective conditions for proliferation of antibiotic resistant bacteria. Comprising a carbon skeleton, antibiotics should be amenable to microbial biodegradation, but this is still largely uncharted territory because of their simultaneous strong toxicity. In this study, we estimated potential antibiotics degradation by and effects on mixed microbial communities at concentrations sufficiently high to allow sensitive detection of biomass growth, but simultaneously, low enough to mitigate their toxic action. We used three different mixed inoculum sources freshly derived from freshwater, activated sludge or soil, and tested a series of 15 antibiotics from different classes at 1 mg C-carbon l−1 dosage. Consistent community growth was observed for freshwater and activated sludge with ampicillin, erythromycin and chloramphenicol, and with sulfomethoxazole for activated sludge, which was accompanied by parent compound disappearance. Community growth could be attributed to a few subclasses of recognized cell types by using supervised machine-learning-based classifiers. Most other tested antibiotics resulted in inhibition of community growth on background assimilable organic carbon, concomitant with altered composition of the resulting communities. We conclude that growth-linked biodegradation of antibiotics at low concentrations may be present among typical environmental microbiota, but for a selected subset only, whereas for the majority of antibiotics negative effects prevail without any sign of productive growth.

Langmuir Monolayer Techniques for the Investigation of Model Bacterial Membranes and Antibiotic Biodegradation Mechanisms.

The amounts of antibiotics of anthropogenic origin released and accumulated in the environment are known to have a negative impact on local communities of microorganisms, which leads to disturbances in the course of the biodegradation process and to growing antimicrobial resistance. This mini-review covers up-to-date information regarding problems related to the omnipresence of antibiotics and their consequences for the world of bacteria. In order to understand the interaction of antibiotics with bacterial membranes, it is necessary to explain their interaction mechanism at the molecular level. Such molecular-level interactions can be probed with Langmuir monolayers representing the cell membrane. This mini-review describes monolayer experiments undertaken to investigate the impact of selected antibiotics on components of biomembranes, with particular emphasis on the role and content of individual phospholipids and lipopolysaccharides (LPS). It is shown that the Langmuir technique may provide information about the interactions between antibiotics and lipids at the mixed film surface (π–A isotherm) and about the penetration of the active substances into the phospholipid monolayer model membranes (relaxation of the monolayer). Effects induced by antibiotics on the bacterial membrane may be correlated with their bactericidal activity, which may be vital for the selection of appropriate bacterial consortia that would ensure a high degradation efficiency of pharmaceuticals in the environment.

Effects of Sulfamethoxazole on Growth and Antibiotic Resistance of A Natural Microbial Community

Diffuse environmental antibiotic and antibiotic resistance gene contamination is increasing human and animal exposure to these emerging compounds with a consequent risk of reduction in antibiotic effectiveness. The present work investigated the effect of the antibiotic sulfamethoxazole (SMX) on growth and antibiotic resistance genes of a microbial community collected from an anaerobic digestion plant fed with cattle manure. Digestate samples were used as inoculum for concentration-dependent experiments using SMX at various concentrations. The antibiotic concentrations affecting the mixed microbial community in terms of growth and spread of resistant genes (sul1, sul2) were investigated through OD (Optical Density) measures and qPCR assays. Moreover, SMX biodegradation was assessed by LC-MS/MS analysis. The overall results showed that SMX concentrations in the range of those found in the environment did not affect the microbial community growth and did not select for antibiotic-resistant gene (ARG) maintenance or spread. Furthermore, the microorganisms tested were able to degrade SMX in only 24 h. This study confirms the complexity of antibiotic resistance spread in real matrices where different microorganisms coexist and suggests that antibiotic biodegradation needs to be included for fully understanding the resistance phenomena among bacteria.

Impacts of biochars on bacterial community shifts and biodegradation of antibiotics in an agricultural soil during short-term incubation

This study investigated the effects of applying different biochars to soil on shifts in the bacterial community, the biodegradation of antibiotics, and their relationships. In total, nine biochars were applied to agricultural soil contaminated with 16 antibiotics. Clustering analysis showed that the responses of bacteria at the genus level to biochars were highly dependent on the biochar feedstock rather than the pyrolysis temperature. Among the antibiotics tested in the study, the biodegradation percentage was lower for tetracyclines (TCs, 6–14%) than sulfonamides (SAs, 8–26%) and quinolones (QLs, 8–24%). For specific individual antibiotics from the same class with similar structures, the high adsorption affinity of soil particles for antibiotics due to hydrophobic interactions (logKow) and electrostatic interactions (pKa) resulted in low biodegradation percentages for antibiotics in the soil. The biodegradation of TCs was affected more by the biochar type (effect size: −10% to 42%) than those of QLs (−26% to 14%) and SAs (−24% to 22%). According to the relationships determined between the bacterial taxonomic composition and biodegradation of antibiotics, Steroidobacter from the phylum Proteobacteria has significant positive correlations with the biodegradation of all SAs (p < 0.01), thereby indicating that Steroidobacter had a high capacity for biodegrading SAs. Significant positive correlations were also detected (p < 0.05) between specific genera (Iamia, Parviterribacter, and Gaiella) from the phylum Actinobacteria and the biodegradation of SAs. No significant positive correlations were found between bacterial genera and the biodegradation percentages for QLs and TCs, possibly due to the specific microorganisms involved in these biodegradation processes. The results in this study provide insights into the biodegradation mechanisms of antibiotics in soil and they may facilitate the development of strategies for the bioremediation of antibiotic-contaminated soil.

Bioaugmentation with immobilized endophytic Penicillium restrictum to improve quorum quenching activity for biofouling control in an aerobic hollow-fiber membrane bioreactor treating antibiotic-containing wastewater

The effects of bioaugmentation with immobilized Penicillium restrictum on the removal efficiency of sulfamethoxazole (SMX), erythromycin (ERY) and tetracycline (TC) antibiotics as well as membrane biofouling was studied using hollow-fiber membrane bioreactor (HF-MBR). Bioaugmentation with P. restrictum led to a significant change in the antibiotic removal efficiency and relative abundance of aerobic microbial community, most probably as a result of its quorum quenching activity. Furthermore, in addition to its role in the increase of SMX and ERY removal efficiencies and the decrease of their sorption on solid phase, bioaugmentation significantly reduced the transmembrane pressure which in turn reduced membrane clogging. The most abundant phyla in sludge and biofilm samples in the presence of P. restrictum were observed to be Proteobacteria, Bacteroidetes and Firmicutes. Differences in bacterial compositions and their specificity in biodegradation of antibiotics in different reactors showed that bacteria were specifically selected under the pressure of antibiotics and growing fungus.

Effects of biochar on biodegradation of sulfamethoxazole and chloramphenicol by Pseudomonas stutzeri and Shewanella putrefaciens: Microbial growth, fatty acids, and the expression quantity of genes

An incubation experiment was conducted to investigate whether different biochar could enhance the biodegradation of sulfamethoxazole (SMX) and chloramphenicol (CAP). During incubation in nutrient medium solution, the degradation efficiencies of SMX by P. stutzeri and S. putrefaciens obtained 61.79% and 68.67% respectively, while CAP was 85.75% and 85.70%. The biodegradation efficiencies of SMX and CAP increased for P. stutzeri cultured with biochar and increased for S. putrefaciens cultured with high-concentration biochar (500, 1,000, 2,000 mg L−1). Additionally, TOC and TN contents were significantly decreased during the biodegradation process. Hence, the effects of biochar on microbial growth, fatty acids and expression genes, biodegradation products were studied. The content of bacteria, saturated fatty acids and expression genes showed a positive correlation with the content of TOC released from biochar, while the biodegradation products would not change when bacteria was cultured with biochar. These indicated that biochar improved the antibiotics biodegradation efficiencies via involvement in the bacterial growth, changing the components of fatty acids, increasing the expression quantity of genes. This research suggests that micro-biological degradation with biochar is a promising technology to treat specific antibiotics in the environment.

Biodegradation of Amoxicillin, Tetracyclines and Sulfonamides in Wastewater Sludge

The removal of antibiotics from the aquatic environment has received great interest. The aim of this study is to examine degradation of oxytetracycline (OTC), tetracycline (TC), chlortetracycline (CTC), amoxicillin (AMO), sulfamethazine (SMZ), sulfamethoxazole (SMX), sulfadimethoxine (SDM) in sludge. Four antibiotic-degrading bacterial strains, SF1 (Pseudmonas sp.), A12 (Pseudmonas sp.), strains B (Bacillus sp.), and SANA (Clostridium sp.), were isolated, identified and tested under aerobic and anaerobic conditions in this study. Batch experiments indicated that the addition of SF1 and A12 under aerobic conditions and the addition of B and SANA under anaerobic conditions increased the biodegradation of antibiotics in sludge. Moreover, the results of repeated addition experiments indicated that the efficiency of the biodegradation of antibiotics using the isolated bacterial strains could be maintained for three degradation cycles. Two groups of potential microbial communities associated with the aerobic and anaerobic degradation of SMX, AMO and CTC in sludge were revealed. Twenty-four reported antibiotics-degrading bacterial genera (Achromobacter, Acidovorax, Acinetobacter, Alcaligenes, Bacillus, Burkholderia, Castellaniella, Comamonas, Corynebacterium, Cupriavidus, Dechloromonas, Geobacter, Gordonia, Klebsiella, Mycobacterium, Novosphingobium, Pandoraea, Pseudomonas, Rhodococcus, Sphingomonas, Thauera, Treponema, Vibrio and Xanthobacter) were found in both the aerobic and anaerobic groups, suggesting that these 24 bacterial genera may be the major antibiotic-degrading bacteria in sludge.

Subsistence and complexity of antimicrobial resistance on a community-wide level.

SummaryThere are a multitude of resistance strategies that microbes can apply to avoid inhibition by antimicrobials. One of these strategies is the enzymatic modification of the antibiotic, in a process generally termed inactivation. Furthermore, some microorganisms may not be limited to the mere inactivation of the antimicrobial compounds. They can continue by further enzymatic degradation of the compounds' carbon backbone, taking nutritional and energetic advantage of the former antibiotic. This driving force to harness an additional food source in a complex environment adds another level of complexity to the reasonably well‐understood process of antibiotic resistance proliferation on a single cell level: It brings bioprotection into play at the level of microbial community. Despite the possible implications of a resistant community in a host and a lurking antibiotic failure, knowledge of degradation pathways of antibiotics and their connections is scarce. Currently, it is limited to only a few families of antibiotics (e.g. β‐lactams and sulfonamides). In this article, we discuss the fluctuating nature of the relationship between antibiotic resistance and the biodegradation of antibiotics. This distinction mainly depends on the genetic background of the microbe, as general resistance genes can be recruited to function in a biodegradation pathway.

Co-metabolism of sulfamethoxazole by a freshwater microalga Chlorella pyrenoidosa

Microalgae-mediated biodegradation of antibiotics has recently gained increased attention from international scientific community. However, limited information is available regarding microalgae-mediated biodegradation of SMX in a co-metabolic system. Here we investigated the biodegradation of sulfamethoxazole (SMX) by five algal species (Pseudokirchneriella subcapitata, Scenedesmus quadricauda, Scenedesmus obliquus, Scenedesmus acuminatus and Chlorella pyrenoidosa), and its transformation pathways by C. pyrenoidosa in a sodium acetate (3 mM) co-metabolic system. The results showed that the highest SMX dissipation (14.9%) was detected by C. pyrenoidosa after 11 days of cultivation among the five tested algal species in the absence of other carbon sources. The addition of sodium acetate (0–8 mM) significantly enhanced the dissipation efficiency of SMX (0.4 μM) from 6.05% to 99.3% by C. pyrenoidosa after 5 days of cultivation, and the dissipation of SMX followed the first-order kinetic model with apparent rate constants (k) ranging from 0.0107 to 0.9811 d−1. Based on the results of mass balance analysis, biodegradation by C. pyrenoidosa was the main mechanism for the dissipation of SMX in the culture medium. Fifteen phase I and phase II metabolites were identified, and subsequently the transformation pathway was proposed, including oxidation, hydroxylation, formylation and side chain breakdown, as well as pterin-related conjugation. The majority of metabolites of SMX were only observed in the culture medium and varied with cultivation time. The findings of the present study showed effective co-metabolism of a sulfonamide by microalgae, and it may be applied in the aquatic environment remediation and wastewater treatment in the future.

Adsorption and biodegradation of sulfamethoxazole and ofloxacin on zeolite: Influence of particle diameter and redox potential

Zeolite played an important role in the removal of antibiotics in constructed wetlands (CWs) due to the high pollution adsorption capacity and tremendous amount of microorganism attachment sites. However, the contribution of adsorption and microbial degradation of this substrates on antibiotic removal, and the degradation difference and mechanism in different redox conditions are still unclear. The adsorption and biodegradation of antibiotics on zeolites with two particle sizes were compared under aerobic, anoxic and anaerobic conditions. The results showed the competitive adsorption strength on powder zeolite was higher than that on mass zeolite. However, the maximum adsorption capacity of sulfamethoxazole (SMX) on powered zeolite was lower than that on massive zeolite, which was mainly due to the destruction of pore structure in powder zeolite. The adsorption effect and mechanism of zeolite to antibiotics can be disrupted by different particle size. Adsorption contributed more than 50% of the total ofloxacin (OFLO) removal, while biodegradation contributed more than 60% of the total SMX removal in our antibiotic removal experiments. In addition, redox conditions can obviously affect removal of antibiotics with the highest removal efficiency for SMX (75%) occurred in aerobic and for OFLO (99.5%) occurred in anaerobic conditions. This phenomenon probably resulted from the different microbial community structures on substrates in anaerobic, aerobic or anoxic under different antibiotic stress. This study can provide strong support for defuzzification of removal mechanism of different antibiotics in CW systems.

Current progress in the adsorption, transport and biodegradation of antibiotics in soil

Antibiotic residues in soil may cause potential risks to human health and soil ecosystems. To avoid these potential risks, comprehensive study of the adsorption, transport and biodegradation of antibiotics in soil is very imperative. This review provided current views about the most recent studies, which have been conducted toward the adsorption, transport and biodegradation of antibiotics in soil. The influencing factors affecting the adsorption behaviors of antibiotics in soil, including the antibiotics properties (e.g., molecular structure, hydrophobicity, polarity, polarizability, and spatial configuration) and the soil characteristics (e.g., soil type, soil pH, coexisting ions, and soil organic matter), were discussed. The effects of fertilizer colloids, porous media, and pH of soil on the transport behaviors of antibiotics were analyzed. The biodegradation of antibiotics in soil were also highlighted by investigating the effects of soil microbiome, soil pH, soil temperature, and interactions between antibiotics. Prospects of antibiotics adsorption, transport and biodegradation were also proposed.

Environmental risk assessment of antibiotics in agroecosystems: ecotoxicological effects on aquatic microbial communities and dissemination of antimicrobial resistances and antibiotic biodegradation potential along the soil-water continuum.

Antibiotics have a wide application range in human and veterinary medicines. Being designed for pharmacological stability, most antibiotics are recalcitrant to biodegradation after ingestion and can be persistent in the environment. Antibiotic residues have been detected as contaminants in various environmental compartments where they cause human and environmental threats, notably with respect to the potential emergence and proliferation of antibiotic-resistant bacteria. An important component of managing environmental risk caused by antibiotics is to understand exposure of soil and water resources to their residues. One challenge is to gain knowledge on the fate of antibiotics in the ecosystem along the soil-water continuum, and on the collateral impact of antibiotics on environmental microorganisms responsible for crucially important ecosystem functions. In this context, the ANTIBIOTOX project aims at studying the environmental fate and impact of two antibiotics of the sulfonamide class of antibiotics, sulfamethazine (SMZ), and sulfamethoxazole (SMX).

Cometabolic biodegradation of cephalexin by enriched nitrifying sludge: Process characteristics, gene expression and product biotoxicity

The nitrifying systems have been reported to be able to biodegrade micropollutants, yet it is still unclear about the cometabolism of ammonia-oxidizing bacteria (AOB) towards micropollutants, in particular their enzyme and transcriptional responses under exposure of micropollutants. This study investigated cometabolic biodegradation of a selected antibiotic, cephalexin (CFX), by an enriched nitrifying culture through a series of batch experiments, together with the assessments of enzymatic activity, key gene expression, and biotoxicity of the degradation products. More than 99% CFX with an initial concentration of 50 μg/L could be removed with the presence of ammonium, while <44% of CFX removal was observed in the absence of ammonium, suggesting the cometabolic degradation of CFX by ammonia-oxidizing bacteria (AOB). After the addition of 50 μg/L CFX, the ammonia oxidizing rate (AOR) decreased from 36.6 to 11.0 mg N/(L·h·g VSS), followed by a slight recovery when CFX concentration decreased to below 8 μg/L. Ammonia monooxygenase (AMO) activity showed a similar trend with that of AOR. The quantitative reverse transcription PCR assay indicated that the expression level of amoA gene was significantly upregulated (up to 3-fold, p < 0.05) due to the addition of CFX, while decreased to the normal level once CFX was degraded, suggesting a mechanism of AOB to neutralize the toxicity of CFX by metabolizing ammonia more effectively. Meanwhile, the biotoxicity test showed the degradation products of CFX did not exhibit any antibacterial impacts in terms of cell viability, compared to the parent compounds. Our finding shed a light on AMO-mediated cometabolic biodegradation of antibiotics in nitrifying cultures.

Influence of high doses of antibiotics on anoxic-aerobic membrane bioreactor in treating solid waste leachate

This study aims to investigate the influence of high antibiotics doses on the efficiency of an anoxic-aerobic membrane bioreactor (ANX-AMBR) in treating solid waste leachate for one-year operation without sludge withdrawal. Two doses of a mixture of norfloxacin (NOR), trimethoprim (TRM), sulfamethoxazole (SMX) and tetracycline (TC), a total of 4 mg L−1 (1 mg L−1 each) and a total of 20 mg L−1 (5 mg L−1 each), had been fed into the ANX-AMBR operated at HRTs of 12 h and 24 h. A high efficiency of organic matter removal could be achieved >90% in all conditions. The highest elimination of NOR, 77%; TRM/SMX/TC, 50–57%, was found under a 12 h-HRT with the antibiotic dose of 4 mg L−1. However, high antibiotic doses gave adverse effects on nitrite reduction-oxidation including antibiotic removal efficiency in the ANX-AMBR. Additionally, the extracellular polymeric substances were produced remarkably in the system after the antibiotic dose of 20 mg L−1. Mass balance analysis suggests that biodegradation was a major role in antibiotics removal in this system. Batch studies showed that the first order kinetics rate constants (k) of antibiotics biodegradation were 0.509–1.573 d−1 of ANX and 0.128–0.516 d−1 of AMBR.

Mechanisms and Influencing Factors of Antibiotic Removal in Sewage Biological Treatment

Antibiotic pollution in the environment is becoming increasingly serious, and the induced antibiotic resistance has become a major threat to human health. A literature review of the antibiotic concentration in the influent and effluent of many sewage treatment plants around the world has shown that current sewage treatment processes do not effectively remove antibiotics. Studies have shown that adsorption and biodegradation are the main ways of removing antibiotics from sewage. The mechanisms of adsorption and the different adsorption extents of diverse antibiotics were analyzed in this paper. Biodegradation of antibiotics in a biological sewage treatment process were analyzed in terms of biodegradability, degrading bacteria, and degradation products. The effects of the operation conditions of biological sewage treatment processes, such as hydraulic retention time, sludge retention time, temperature, and process selection (conventional activated sludge, membrane bioreactor, or biological nitrogen removal processes), on adsorption and biodegradation pathways as well as on the removal efficiency of antibiotics are also discussed. The effects of bacterial composition, growth substrate, and coexisting micro-pollutants on the fate of antibiotics in biological wastewater treatment require more in-depth research.

Degradation of oxytetracycline (OTC) and nitrogen conversion characteristics using a novel strain

A high residue of oxytetracycline (OTC) and high concentrations of ammonia and nitrate remain in aquaculture wastewater, and it is therefore important to identify a heterotrophic nitrification-aerobic denitrifying bacterium that also provides highly efficient OTC degradation for use in practical applications. In this study, a novel OTC degrading strain with aerobic denitrification, Ochrobactrum sp KSS10, was isolated and identified, and the heterotrophic nitrification and denitrification processes were investigated under aerobic conditions. Results suggest that 97.41% of ammonia and 82.86% of nitrate were removed by strain KSS10 within 112 h. Interestingly, a large amount of nitrite accumulation was observed in the middle stage, but this finally disappeared, and a nitrogen balance analysis showed that approximately 75% of nitrogen was lost in the form of gas products. The relationship between OTC degradation and the nitrogen conversion effect was then explored, and a maximum OTC reduction rate of 0.012 mg·L−1·h−1 and removal rates of 0.848 mg·L−1·h−1 of NH4+-N and 1.189 mg·L−1·h−1 of TN were observed from 24 h to 104 h, respectively. The nitrate concentration decreased sharply from 40 h to 80 h with a maximum removal rate of 1.967 mg·L−1·h−1 of NO3−-N. Furthermore, dissolved organic matter (DOM) and chemical functional groups were detected to analyze the main mechanisms involved in the degradation of OTC and nitrogen (N) removal. In conclusion, the results show that the use of strain KSS10 in aquaculture wastewater can potentially be used in the biodegradation of antibiotics and nitrogen removal.

Distribution, dynamics and determinants of antibiotics in soils in a peri-urban area of Yangtze River Delta, Eastern China

Antibiotics are increasingly recognized as anthropogenic contaminants in soils, and they can persist through a complex vicious cycle of transformation and bioaccumulation. In this study, we quantified 11 quinolones (QNs), 5 sulfonamides (SAs), 5 macrolides (MLs), and 4 tetracyclines (TCs) in soils at three soil layers (0–10, 10–20, 20–40 cm) in a typical peri-urban catchment in the Yangtze River Delta, Eastern China. The results showed that total antibiotic levels were significantly higher in cropland topsoil (p < 0.05) compared to orchards and forests (p < 0.05). Moreover, a significant seasonal variation for antibiotic concentrations in croplands’ topsoil were observed in the summer (50.59 ± 84.55 ng/g) and winter (112.44 ± 140.58 ng/g). Chlortetracycline (15.30 ± 45.44 ng/g), enrofloxacin (0.43 ± 0.93 ng/g), sulfamethazine (0.05 ± 0.02 ng/g) and clarithromycin (0.03 ± 0.03 ng/g) were detected with the highest frequencies within TCs, QNs, SAs, and MLs, respectively. Concentrations of TCs, QNs, and SAs decreased with increasing soil depth. The concentrations of TCs, QNs, and SAs were significantly affected by the intensity of human activities. According to the results of redundancy analysis (RDA), anthropogenic effects on the distribution of antibiotics in soils in winter were so strong that they dwarfed the effects of environmental factors. In summer, human activities and their interactions with environmental factors were the dominant contributors to variations in soil antibiotics. In addition, the results of RDA suggested that soil pH and organic matter closely correlated with the levels of antibiotics, and Actinobacteria was the predominant contributor to the biodegradation of antibiotics in this study area.

Elimination of Isoxazolyl-Penicillins antibiotics in waters by the ligninolytic native Colombian strain Leptosphaerulina sp. considerations on biodegradation process and antimicrobial activity removal

In this work, Leptosphaerulina sp. (a Colombian native fungus) significantly removed three Isoxazolyl-Penicillin antibiotics (IP): oxacillin (OXA, 16000 μg L−1), cloxacillin (CLX, 17500 μg L−1) and dicloxacillin (DCX, 19000 μg L−1) from water. The biological treatment was performed at pH 5.6, 28 °C, and 160 rpm for 15 days. The biotransformation process and lack of toxicity of the final solutions (antibacterial activity (AA) and cytotoxicity) were tested. The role of enzymes in IP removal was analysed through in vitro studies with enzymatic extracts (crude and pre-purified) from Leptosphaerulina sp., commercial enzymes and enzymatic inhibitors. Furthermore, the applicability of mycoremediation process to a complex matrix (simulated hospital wastewater) was evaluated. IP were considerably abated by the fungus, OXA was the fastest degraded (day 6), followed by CLX (day 7) and DCX (day 8). Antibiotics biodegradation was associated to laccase and versatile peroxidase action. Assays using commercial enzymes (i.e. laccase from Trametes versicolor and horseradish peroxidase) and inhibitors (EDTA, NaCl, sodium acetate, manganese (II) ions) confirmed the significant role of enzymatic transformation. Whereas, biomass sorption was not an important process in the antibiotics elimination. Evaluation of AA against Staphylococcus aureus ATCC 6538 revealed that Leptosphaerulina sp. also eliminated the AA. In addition, the cytotoxicity assay (MTT) on the HepG2 cell line demonstrated that the IP final solutions were non-toxic. Finally, Leptosphaerulina sp. eliminated OXA and its AA from synthetic hospital wastewater at 6 days. All these results evidenced the potential of Leptosphaerulina sp. mycoremediation as a novel environmentally friendly process for the removal of IP from aqueous systems.

Potential for natural and enhanced attenuation of sulphanilamide in a contaminated chalk aquifer

Understanding antibiotic biodegradation is important to the appreciation of their fate and removal from the environment. In this research an Isotope Ratio Mass Spectrometry (IRMS) method was developed to evaluate the extent of biodegradation of the antibiotic, sulphanilamide, in contaminated groundwater. Results indicted an enrichment in δ13C of 8.44‰ from −26.56 (at the contaminant source) to −18.12‰ (300m downfield of the source). These results confirm reductions in sulphanilamide concentrations (from 650 to 10mg/L) across the contaminant plume to be attributable to biodegradation (56%) vs. other natural attenuation processes, such as dilution or dispersion (42%). To understand the controls on sulphanilamide degradation ex-situ microcosms assessed the influence of sulphanilamide concentration, redox conditions and an alternative carbon source. Results indicated, high levels of anaerobic capacity (~50% mineralisation) to degrade sulphanilamide under high (263mg/L), moderate (10mg/L) and low (0.02mg/L) substrate concentrations. The addition of electron acceptors; nitrate and sulphate, did not significantly enhance the capacity of the groundwater to anaerobically biodegrade sulphanilamide. Interestingly, where alternative carbon sources were present, the addition of nitrate and sulphate inhibited sulphanilamide biodegradation. These results suggest, under in-situ conditions, when a preferential carbon source was available for biodegradation, sulphanilamide could be acting as a nitrogen and/or sulphur source. These findings are important as they highlight sulphanilamide being used as a carbon and a putative nitrogen and sulphur source, under prevailing iron reducing conditions.

Quantification of residual antibiotics in cow manure being spread over agricultural land and assessment of their behavioral effects on antibiotic resistant bacteria

Antibiotic resistant bacteria (ARB) in livestock manure used as fertilizer and spread over agriculture land, may pose a threat to the health of humans. Considering this, the concentrations of tetracycline (TC), oxytetracycline (OTC), and sulfathiazole (STZ) in the surface soil were quantified using LC-MS. These antibiotics have been used in livestock and are found in fertilizer produced from livestock excretions. Species of ABR were identified using 16S rDNA. Soil samples were collected at depths of 0, 7, and 15 cm from farmland in Incheon (South Korea). In the surface soil, three compounds were detected: TC (17.74 μg/kg), OTC (0.78 μg/kg), and STZ (0.23 μg/kg). However, except for STZ, antibiotics were not detected in the deeper samples. Overall, TC can form a chelated complex with cations, which consequently enhances its adsorption to the organic matter and metals in soil. This property can significantly reduce the mobility of TC (to lower than that of STZ). The result of 16S rDNA gene analysis indicated that Pseudomonas spp., Arthrobacter spp., and Rhodococcus spp. showed persistent resistance to the three antibiotics tested. DNA quantification results revealed strong resistance of Pseudomonas spp. to STZ, whereas Arthrobacter spp. and Rhodococcus spp. had resistance to TC and OTC. Antibiotics biodegradation suggested ability of ARB to grow in soil samples in presence of residual antibiotics during 13 days incubation. The concentrations of STZ, TC, and OTC reduced as much as 23.53, 35.60 and 66.88%, respectively.