Increasing Tetracycline Concentrations Research Articles

Effective degradation of tetracycline via persulfate activation using silica-supported zero-valent iron: process optimization, mechanism, degradation pathways and water matrices.

Pure zero-valent iron (ZVI) was supported on silica and starch to enhance the activation of persulfate (PS) for tetracycline degradation. The synthesized catalysts were characterized by microscopic and spectroscopic methods to assess their physical and chemical properties. High tetracycline removal (67.55%) occurred using silica modified ZVI (ZVI-Si)/PS system due to the improved hydrophilicity and colloidal stability of ZVI-Si. Incorporating light into the ZVI-Si/PS system improved the degradation performance by 9.45%. Efficient degradation efficiencies were recorded at pH 3-7. The optimum operating parameters determined by the response surface methodology were PS concentration of 0.22mM, initial tetracycline concentration of 10mg/L, and ZVI-Si dose of 0.46g/L, respectively. The rate of tetracycline degradation declined with increasing tetracycline concentration. The degradation efficiencies of tetracycline were 77%, 76.4%, 75.7%, 74.5%, and 73.75% in five repetitive runs at pH 7, 20mg/L tetracycline concentration, 0.5g/L ZVI-Si dose, and 0.1mM PS concentration. The degradation mechanism was explained, and sulfate radicals were the principal reactive oxygen species. The degradation pathway was proposed based on liquid chromatography-mass spectroscopy. Tetracycline degradation was favorable in distilled and tap water. The ubiquitous presence of inorganic ions and dissolved organic matter in the lake, drain, and seawater matrices interfered with the tetracycline degradation. The high reactivity, degradation performance, stability, and reusability of ZVI-Si substantiate the potential practical application of this material for the degradation of real industrial effluents.

Tetracycline-Induced Release and Oxidation of As(III) Coupled with Concomitant Ferrihydrite Transformation

Cocontamination with tetracycline (TC) and arsenic (As) is very common in paddy fields. However, the process and underlying mechanism of arsenite (As(III)) transformation on iron mineral surfaces in the presence of antibiotic contaminants remain unclear. In this study, the release and oxidation of As(III) on ferrihydrite (Fh) surfaces and Fh transformation in the presence of TC under both aerobic and anaerobic conditions were investigated. Our results indicated that the TC-induced reductive dissolution of Fh (Fe(II) release) and TC competitive adsorption significantly promote the release of As, especially under anaerobic conditions. The release of As was increased with increasing TC concentration, whereas it decreased with increasing pH. Interestingly, under both aerobic and anaerobic conditions, the addition of TC enhanced the oxidation of As(III) by Fh and induced the partial transformation of Fh to lepidocrocite. Under aerobic conditions, the adsorbed Fe(II) activated the production of reactive oxygen species (·OH and 1O2) from dissolved O2, with Fe(IV) being responsible for As(III) oxidation. Under anaerobic conditions, the abundant oxygen vacancies of Fh affected the oxidation of As(III) during Fh recrystallization. Thus, this study provided new insights into the role of TC on the migration and transformation of As coupled with Fe in soils.

Response of immobilized denitrifying bacterial consortium to tetracycline exposure

Tetracycline (TC) as one of the most widely used antibiotics commonly exists in aquaculture tail water and piggery wastewater, causing risks to human. However, the response of immobilized anaerobic denitrifying bacterial consortium to TC exposure lacks systematic research. In this study, the denitrification performance and the compositional shift of extracellular polymeric substances (EPS) and microbial community under TC stress were investigated. The inhabitation effect of TC on nitrate reduction of the immobilized bacterial consortium became evident at high concentrations (50 mg/L and 100 mg/L). Nitrite reduction was more sensitively inhibited than nitrate reduction. The inhabitation effect was mainly due to the fact that TC damaged cell membranes and subsequently effect the intracellular enzymes activities relating to denitrification (NAR and NIR activities). About 50% of TC can be removed by the immobilized bacterial consortium under all tested TC concentrations. Three-dimensional excitation-emission matrix (3D-EEM) results implied that the tryptophan like substances of EPS were obviously quenched with increasing TC concentration. EPS played an important role in TC removal. The denitification performance of the immobilized bacterial consortium under TC stress was attributed to the genera Paraccoccus, Pseudoxanthomonas, Diaphorobacter and Pseudomonas. Initial TC concentration obviously affected the microbial communities. This study may facilitate the management of aquaculture tail water and piggery wastewater contaminated with nitrate and antibiotics.

Novel sodium bicarbonate activation of cassava ethanol sludge derived biochar for removing tetracycline from aqueous solution: Performance assessment and mechanism insight

NaHCO3 was used as a novel activator to produce cassava ethanol sludge-based biochar. The NaHCO3-activated biochar showed superior adsorption capacity for tetracycline (154.45 mg/g) than raw biochar (34.04 mg/g). Orthogonal experiments confirmed the optimal preparation conditions of biochar. Increasing adsorbent dosage and temperature facilitated tetracycline removal. The maximum removal was 92.60% at pH = 3.0. Calcium ions and alkalinity decreased tetracycline removal. The time for attaining equilibrium was extended with increasing tetracycline concentration, but the equilibrium could be completed within 24 h. Langmuir model fitted the equilibrium data well. Kinetics process followed the Elovich model. The adsorption rate was controlled by both intraparticle and liquid film diffusion and the process was endothermic and spontaneous. The electrostatic attraction, hydrogen bonding, π-π interactions, and pore-filling were involved in the adsorption mechanism. The findings may provide an underlying guide for sludge disposal and removal of tetracycline from wastewater in practical application.

Impact of Tetracycline Antibiotic on the Transcriptional Expression of Tetracycline Resistance Genes in Shigella flexneri

The tetracycline (TC) antibiotic has been widely found in different environmental matrices. The tetracycline resistant bacterium (TRB) of Shigella flexneri was screened and purified from activated sludge, and was then used to study the impact of TC stress on the gene abundances and expression levels of TC resistance genes (TC-ARGs), including tetC, tetO, and tetX, which were respectively quantified by quantitative PCR and reverse transcriptional PCR. Correlations between the TC concentration and gene abundances of TC-ARGs and their expression levels were discussed. The results showed that TC stress had an inhibiting effect on the growth of Shigella flexneri during the entire culture cycle (24 h) and that the growth rate of the bacterial concentration decreased with increasing TC concentration. However, less impact on the gene abundance of TC-ARGs was found. TC stress could promote the expression of TC-ARGs in Shigella flexneri, and the expression levels of tetC, tetO, and tetX genes first increased and then decreased. The correlation results indicated that no significant correlation was observed between the TC concentration and gene abundances of TC-ARGs and their expression levels. Nevertheless, the gene abundances of tetC and tetO were significantly correlated with their expression levels, thus indicating that they can be used to evaluate and assess expression levels to a certain extent.

World within world: Intestinal bacteria combining physiological parameters to investigate the response of Metaphire guillelmi to tetracycline stress

Due to the abusive usage of antibiotics in animal husbandry, a large amount of residual antibiotics has been released into the environment, therein posing great threat against both environment security and public health. Therefore, it is of great significance to investigate the toxicity of antibiotics on the widely-applied bioindicator-earthworm. In this work, the physiological parameters and the intestinal bacteria community of Metaphire guillelmi were monitored simultaneously to evaluate their sensitivity to the tetracycline (TC) exposure. As expected, the antioxidant enzyme activity and coelomocyte apoptosis acted fairly well as biomarkers for the TC toxicity. In contrast, the intestinal bacteria of Metaphire guillelmi responded varyingly to different TC doses. When TC concentration increased from 0 to 35.7 μg cm−2, the percentage of the Proteobacteria phylum declined significantly from 85.5% to 34.4%, while the proportions of the Firmicutes, Planctomycetes and Atinomycete phyla clearly increased (p < 0.05). Meanwhile, the levels of TC resistance genes tetA, tetC, and tetW increased with the increasing TC concentration, in contrast to the declined abundance in denitrifying genes nirS and nosZ (p < 0.05). By analyzing the correlation between the antioxidant enzyme activity and the dominant intestinal bacteria in the worm gut, it is interesting to found that the four dominant bacteria genera Mesorhizobium, Aliihoeflea, Romboutsia, and Nitrospira are the promising bioindicator of TC stress due to their sensitive response. This work shed novel light on evaluating the ecotoxicological risks posed by residual TC in environment by using a combination of physiological parameters and intestinal bacterial activity in earthworms.

Effect of Tetracycline Antibiotic on Abundance and Transcriptional Expression Level of Tetracycline Resistance Genes in Activated Sludge

Tetracycline resistant bacteria (TRB) screened from activated sludge were used to study the effect of tetracycline (TC) antibiotic on the transcriptional expression of tetracycline resistance genes (TC-ARGs). The gene abundances of seven TC-ARGs including tetA, tetC, tetG, tetM, tetO, tetW, and tetX, as well as their expression levels, were quantified by quantitative PCR (qPCR) and reverse transcriptional PCR (RT-PCR). The correlations between TC concentrations and gene abundance of TC-ARGs and their expression levels were discussed. The results showed that the gene abundances of tetA, tetG, and tetW generally increased with increasing TC exposure concentrations during the entire culture cycle, whereas other TC-ARGs fluctuated greatly. The impact of TC stress on the transcriptional expression level of different TC-ARGs varied to a great extent. The gene expression of tetA was relatively stable and exhibited an upregulated trend with increasing TC concentrations. When the TC concentration was 100 mg·L-1, the upregulation of tetA expression was as high as 5.3-fold compared with the control. Under short-term TC stress (one day), the transcriptional expression level was upregulated with increasing TC concentration. The correlation results showed that gene abundances of tetA and tetW correlated significantly with their respective expression levels, indicating that they can evaluate expression levels to a certain extent, which can further mirror functional activities and environmental risks.

Increasing tetracycline concentrations on the performance and communities of mixed microalgae-bacteria photo-bioreactors

This study investigated the impact of varying concentrations of tetracycline on the performance of mixed microalgae-bacteria photo-bioreactors. Photo-bioreactors were assessed for their ability to remove carbon dioxide (CO2) from the biogas of anaerobic membrane bioreactor (anMBR), and nutrients from the anaerobic effluent. The varying concentrations of tetracycline had no impact on the removal of CO2 from biogas. 29% v/v of CO2 was completely removed to generate >20% v/v of oxygen (O2) in all reactors. Removal of nutrients and biomass was not affected at low concentrations of tetracycline (≤150μg/L), but 20mg/L of tetracycline lowered the biomass generation and removal efficiencies of phosphate. Conversely, high chlorophyll a and b content was observed at 20mg/L of tetracycline. High tetracycline level had no impact on the diversity of 18S rRNA gene-based microalgal communities but adversely affected the 16S rRNA gene-based microbial communities. Specifically, both Proteobacteria and Bacteroidetes phyla decreased in relative abundance but not phylum Chloroplast. Additionally, both nitrogen-fixing (e.g. Flavobacterium, unclassified Burkholderiales and unclassified Rhizobiaceae) and denitrifying groups (e.g. Hydrogenophaga spp.) were significantly reduced in relative abundance at high tetracycline concentration. Phosphate-accumulating microorganisms, Acinetobacter spp. and Pseudomonas spp. were similarly reduced upon exposure to high tetracycline concentration. Unclassified Comamonadaceae, however, increased in relative abundance, which correlated with an increase in the abundance of tetracycline resistance genes associated with efflux pump mechanism. Overall, the findings demonstrate that antibiotic concentrations in municipal wastewaters will not significantly affect the removal of nutrients by the mixed microalgae-bacteria photo-bioreactors. However, utilizing such photo-bioreactors as a polishing step for anMBRs that treat wastewaters with high tetracycline concentration may not be effective as evidenced from the lower nutrient removal and occurrence of antibiotic resistance genes.

Performance and microbial community variations of anaerobic digesters under increasing tetracycline concentrations

The impact of different concentrations of tetracycline on the performance of anaerobic treatment was evaluated. Results revealed that for all of the tested tetracycline concentrations, no major sustained impact on methane production was observed. Instead, a significant increase in propionic acid was observed in the reactor subjected to the highest concentration of tetracycline (20 mg/L). Microbial community analyses suggest that an alternative methanogenic pathway, specifically that of methanol-utilizing methanogens, may be important for ensuring the stability of methane production in the presence of high tetracycline concentrations. In addition, the accumulation of propionate was due to an increase in volatile fatty acids (VFA)-producing bacteria coupled with a reduction in propionate utilizers. An increase in the abundance of tetracycline resistance genes associated with ribosomal protection proteins was observed after 30 days of exposure to high concentrations of tetracycline, while other targeted resistance genes showed no significant changes. These findings suggest that anaerobic treatment processes can robustly treat wastewater with varying concentrations of antibiotics while also deriving value-added products and minimizing the dissemination of associated antibiotic resistance genes.

Minimal selective concentrations of tetracycline in complex aquatic bacterial biofilms

Selection pressure generated by antibiotics released into the environment could enrich for antibiotic resistance genes and antibiotic resistant bacteria, thereby increasing the risk for transmission to humans and animals. Tetracyclines comprise an antibiotic class of great importance to both human and animal health. Accordingly, residues of tetracycline are commonly detected in aquatic environments. To assess if tetracycline pollution in aquatic environments promotes development of resistance, we determined minimal selective concentrations (MSCs) in biofilms of complex aquatic bacterial communities using both phenotypic and genotypic assays. Tetracycline significantly increased the relative abundance of resistant bacteria at 10μg/L, while specific tet genes (tetA and tetG) increased significantly at the lowest concentration tested (1μg/L). Taxonomic composition of the biofilm communities was altered with increasing tetracycline concentrations. Metagenomic analysis revealed a concurrent increase of several tet genes and a range of other genes providing resistance to different classes of antibiotics (e.g. cmlA, floR, sul1, and mphA), indicating potential for co-selection. Consequently, MSCs for the tet genes of ≤1μg/L suggests that current exposure levels in e.g. sewage treatment plants could be sufficient to promote resistance. The methodology used here to assess MSCs could be applied in risk assessment of other antibiotics as well.

Relation between tetR and tetA expression in tetracycline resistant Escherichia coli.

BackgroundTetracyclines are among the most used antibiotics in livestock worldwide. Resistance is widely disseminated in Escherichia coli, where it is generally mediated by tetracycline efflux pumps, such as TetA. Expression of tetracycline efflux pumps is tightly controlled by the repressor TetR, which has been shown to be tetracycline-responsive at sub-MIC tetracycline concentrations. The objective of this study was to investigate the effects of increasing tetracycline concentrations on the growth of TetA-producing E. coli, and to determine how expression of tetA and tetR related to each other in different growth phases in the presence of tetracycline.ResultsA tetracycline resistant E. coli strain containing tetA and tetR on the chromosome was constructed and cultured in the presence of increasing concentrations of tetracycline. Expression of tetR and tetA was measured at four time points in different growth phases by quantitative real-time PCR. The TetA-producing E. coli exhibited prolonged lag phase with increasing concentrations of tetracycline, while expression of tetA and tetR increased and decreased, respectively, with increasing tetracycline concentration. The levels of tetA and tetR mRNA varied depending on growth phase, resulting in a gradual decrease of the tetA/tetR ratio from approximately 4 in the lag phase to approximately 2 in the stationary phase.ConclusionThis study shows that the expression of tetR and tetA is tetracycline concentration- and growth phase-dependent, contributing to improved understanding of the relationships between E. coli growth, tetracycline exposure and expression of tetracycline resistance.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-016-0649-z) contains supplementary material, which is available to authorized users.

Pharmacodynamic modelling of in vitro activity of tetracycline against a representative, naturally occurring population of porcine Escherichia coli.

BackgroundThe complex relationship between drug concentrations and bacterial growth rates require not only the minimum inhibitory concentration but also other parameters to capture the dynamic nature of the relationship. To analyse this relationship between tetracycline concentration and growth of Escherichia coli representative of those found in the Danish pig population, we compared the growth of 50 randomly selected strains. The observed net growth rates were used to describe the in vitro pharmacodynamic relationship between drug concentration and net growth rate based on Emax model with three parameters: maximum net growth rate (αmax); concentration for a half-maximal response (Emax); and the Hill coefficient (γ).ResultsThe net growth rate in the absence of antibiotic did not differ between susceptible and resistant isolates (P = 0.97). The net growth rate decreased with increasing tetracycline concentrations, and this decline was greater in susceptible strains than resistant strains. The lag phase, defined as the time needed for the strain to reach an OD600 value of 0.01, increased exponentially with increasing tetracycline concentration. The pharmacodynamic parameters confirmed that the alpha_{max} between susceptible and resistant strains in the absence of a drug was not different. EC50 increased linearly with MIC on a log–log scale, and γ was different between susceptible and resistant strains.ConclusionsThe in vitro model parameters described the inhibition effect of tetracycline on E. coli when strains were exposed to a wide range of tetracycline concentrations. These parameters, along with in vivo pharmacokinetic data, may be useful in mathematical models to predict in vivo competitive growth of many different strains and for development of optimal dosing regimens for preventing selection of resistance.

Tetracycline modifies competitive interactions in experimental microcosms containing bacteria isolated from freshwater.

Interspecific microbial interactions are important for community structure, function and evolution, but it is not fully understood how interactions among bacterial species are influenced by some types of abiotic environmental variation, such as exposure to antibiotics. We tested for the effect of an antibiotic, tetracycline, on interspecific interactions in vitro among four species of aquatic bacteria isolated from European water bodies. Interactions among species in experimental microcosms containing artificial lake water (ALW) supplemented with glucose were largely competitive, as detected by comparing mixed cultures to pure cultures of their constituent species. Sublethal concentrations of tetracycline changed the relative competitive abilities of different species and revealed considerable variation in antibiotic sensitivity, but did not reduce the average strength of competition. Interspecific interactions at a given concentration were largely predictable from growth in pure cultures and indirect interactions with other species. These results suggest that antibiotics such as tetracycline may have important consequences for interactions among bacterial species, but in our experiments this was because species varied in their capacities for growth in the presence of tetracycline, rather than reduced competition at increasing tetracycline concentrations.

Electrochemical aptasensor for the detection of tetracycline with multi-walled carbon nanotubes amplification

Herein, we present a simple electrochemical tetracycline (TET) aptasensor with multi-walled carbon nanotubes (MWCNTs) modification. MWCNTs were dropped on the glassy carbon electrode (GCE) to immobilize the anti-TET aptamer and to construct the aptasensor. The stepwise assembly process of the aptasensor was characterized by cyclic voltammetry. Results demonstrated that the peak currents of Fe(CN)63−/Fe(CN)64− redox pair decreased due to the formation of anti-TET/TET complexes on the modified electrode. The optimization of the loading amount of MWCNTs, the incubating conditions of aptamer and the detection time of TET were investigated in details. Under optimal conditions, the peak currents change obtained by DPV increased linearly with the increasing TET concentrations in the range from 1×10−8M to 5×10−5M with a linear coefficiency of 0.995. This electrochemical aptasensor has a detection limit of 5×10−9M and was successfully applied to the determination of TET in spiked milk samples.

Aqueous photolysis of tetracycline and toxicity of photolytic products to luminescent bacteria

The extensive utilization of antibiotics in the pharmaceutical therapies and agricultural husbandry has led to the worldwide pollution in environments. In this study the photolysis behaviors of tetracycline (TC) and toxicity of its degradation products were investigated. The results showed that TC photolysis followed first-order kinetics. The photolysis rate was found to be dependent on the initial TC concentration and increasing TC concentration from 10 to 40 mg l −1 led to the decrease of the photolysis rate constant from 0.0045 min −1 to 0.0014 min −1. TC photolysis was highly pH-dependent and strongly enhanced at high pH value. Markedly elevated TC photolysis was also observed in the presence of nitrate and dissolved organic matter. Upon irradiation for 300 min, only 15% reduction of total organic carbon (TOC) occurred in spite of quick conversion of 73% of TC, suggesting that a majority of TC transformed into intermediate products without complete mineralization. The intermediate products from TC photolysis were analyzed using high-performance liquid chromatography–electrospray ionisation-mass spectrometry (HPLC–ESI-MS) and the main photolysis products of TC were proposed. The toxicity of the photolysis compounds was evaluated using luminescent bacterium, and the results revealed that the toxicity increased with irradiation, indicative of a higher adversity risk of the degradation products of TC on bacteria upon photolysis.

Mechanism of DNA strand breakage induced by photosensitized tetracycline–Cu(II) complex

Tetracyclines (TCs) in combination with Cu(II) ions exhibited significant DNA damaging potential vis a vis tetracyclines per se. Interaction of tetracyclines with DNA resulted in alkylation at N-7 and N-3 positions of adenine and guanine bases, and caused destabilization of DNA secondary structure. Significant release of acid-soluble nucleotides from tetracycline-modified DNA upon incubation with S 1 nuclease ascertained the formation of single stranded regions in the DNA. Also, the treatment of tetracycline-modified DNA with 0.1 and 0.5 M NaOH resulted in 62 and 76% hydrolysis compared to untreated control. Comparative alkaline hydrolysis of DNA modified with tetracycline derivatives showed differential DNA damaging ability in the order as DOTC>DMTC>TC>OTC>CTC. Addition of Cu(II) invariably augmented the extent of tetracycline-induced DNA damage. The alkaline unwinding assay clearly demonstrated the formation of approximately six strand breaks per unit DNA at 1:10 DNA nucleotide/TC molar ratio in the presence of 0.1 mM Cu(II) ions. At a similar Cu(II) concentration, a progressive transformation of covalently closed circular (CCC) (form-I) plasmid pBR322 DNA to forms-II and -III was noticed with increasing tetracycline concentrations. The results obtained with the free-radical quenchers viz. mannitol, thiourea, sodium benzoate and superoxide dismutase (SOD) suggested the involvement of reactive oxygen species in the DNA strand breakage. It is concluded that the tetracycline–Cu(II)-induced DNA damage occurs due to (i) significant binding of tetracycline and Cu(II) with DNA, (ii) methyl group transfer from tetracycline to the putative sites on nitrogenous bases, and (iii) metal ion catalyzed free-radical generation in close vicinity of DNA backbone upon tetracycline photosensitization. Albeit, the DNA alkylation and strand cleavage are repairable lesions, but any defect in the critical repair pathway may augment the damage accumulation and mutagenesis.