Bioguided Fractionation of Local Plants against Matrix Metalloproteinase9 and Its Cytotoxicity against Breast Cancer Cell Models: In Silico and In Vitro Study (Part II).

In our previous work, the partitions (1 mg/mL) of Ageratum conyzoides (AC) aerial parts and Ixora coccinea (IC) leaves showed inhibitions of 94% and 96%, respectively, whereas their fractions showed IC50 43 and 116 µg/mL, respectively, toward Matrix Metalloproteinase9 (MMP9), an enzyme that catalyzes a proteolysis of extracellular matrix. In this present study, we performed IC50 determinations for AC n-hexane, IC n-hexane, and IC ethylacetate partitions, followed by the cytotoxicity study of individual partitions against MDA-MB-231, 4T1, T47D, MCF7, and Vero cell lines. Successive fractionations from AC n-hexane and IC ethylacetate partitions led to the isolation of two compounds, oxytetracycline (OTC) and dioctyl phthalate (DOP). The result showed that AC n-hexane, IC n-hexane, and IC ethylacetate partitions inhibit MMP9 with their respective IC50 as follows: 246.1 µg/mL, 5.66 µg/mL, and 2.75 × 10-2 µg/mL. Toward MDA-MB-231, 4T1, T47D, and MCF7, AC n-hexane demonstrated IC50 2.05, 265, 109.70, and 2.11 µg/mL, respectively, whereas IC ethylacetate showed IC50 1.92, 57.5, 371.5, and 2.01 µg/mL, respectively. The inhibitions toward MMP9 by OTC were indicated by its IC50 18.69 µM, whereas DOP was inactive. A molecular docking study suggested that OTC prefers to bind to PEX9 rather than its catalytic domain. Against 4T1, OTC showed inhibition with IC50 414.20 µM. In conclusion, this study furtherly supports the previous finding that AC and IC are two herbals with potential to be developed as triple-negative anti-breast cancer agents.

Quantification of Veterinary Antibiotics in Pig and Poultry Feces and Liquid Manure as a Non-Invasive Method to Monitor Antibiotic Usage in Livestock by Liquid Chromatography Mass-Spectrometry.

Antibiotics are active substances frequently used to treat and prevent diseases in animal husbandry, especially in swine and poultry farms. The use of manure as a fertilizer may lead to the dispersion of antibiotic residue into the environment and consequently the development of antibiotic-resistant bacteria. Most pharmaceutical active ingredients are excreted after administration, in some cases up to 90% of the consumed dose can be found in the feces and/or urine as parent compound. Therefore, due to antibiotic metabolism their residues can be easily detected in manure. This article describes a method for simultaneous analysis of ciprofloxacin, chlortetracycline, doxycycline, enrofloxacin, lincomycin, oxytetracycline, tetracycline, tiamulin, trimethoprim and tylosin in feces, liquid manure and digestate. Antibiotics were extracted from the different matrices with McIlvaine-Na2EDTA buffer solution and the extract was purified by the use two techniques: d-SPE and SPE (Strata-X-CW cartridges) and final eluent was analyzed by LC-MS and LC-MS/MS. The European Commission Decision 2002/657/EC was followed to conduct the validation of the method. Recoveries obtained from spiked pig and poultry feces and liquid manures samples ranged from 63% to 93% depending on analytes. The analysis of 70 samples (feces, liquid manure and digestate) revealed that 18 samples were positive for the presence of doxycycline, oxytetracycline, tetracycline, chlortetracycline, enrofloxacin, tiamulin and lincomycin. The results obtained in the presented study demonstrated that animal feces can be used as a non-invasive method detection antibiotic usage in animal production.

Low-cost field production of biochars and their properties.

Biochar has been intensively investigated for carbon sequestration, soil fertility enhancement, and immobilization of heavy metals and organic pollutants. Large-scale use of biochar in agricultural production and environmental remediation, however, has been constrained by its high cost. Here, we demonstrated the production of low-cost biochar ($20/ton) in the field from Robinia pseudoacacia biowaste via a combined aerobic and oxygen-limited carbonization process and a fire-water-coupled method. It involved aerobic combustion at the outer side of biomass, oxygen-limited pyrolysis in the inner core of biomass, and the termination of the carbonization by water spray. The properties of biochar thus produced were greatly affected by exposure time (the gap between a burning char fell to the ground and being extinguished by water spray). Biochar formed by zero exposure time showed a larger specific surface area (155.77 m2/g), a higher carbon content (67.45%), a lower ash content (15.38%), and a higher content of carboxyl and phenolic-hydroxyl groups (1.74 and 0.86 mol/kg, respectively) than biochars formed with longer exposure times (5-30 min). Fourier-transform infrared spectroscopic (FTIR) spectra indicated that oxygen-containing functional groups of biochar played a role in Cd and oxytetracycline sorption though a quantitative relationship could not be established as the relative contribution of carbon and ash moieties of biochar to the sorption was unknown. Outcomes from this research provide an option for inexpensive production of biochar to support its use as a soil amendment in developing countries.

Evaluation of antibiotic oxytetracycline removal in water using a gas phase dielectric barrier discharge plasma.

Degradation of oxytetracycline (OTC), a primary member of antibiotics in water, was performed by a gas phase dielectric barrier discharge (GPDBD) plasma reactor. The influences of operation conditions including applied voltages, air bubbling rates, initial OTC concentrations and initial pH values on OTC abatement were investigated respectively. The results showed that the decontamination process can be fitted by first order kinetics, and the removal ratio and rate were affected obviously by those parameters. After 20 min of discharge treatment, approximately 93.4% of OTC was removed under the experimental conditions: applied voltage of 7.5 kV, air flow rate of 1.0 L/min, initial OTC concentration of 100 mg/L, and initial pH of 5.0. In addition, TOC and COD removal efficiency reached 43.0% and 73.7% at the original pH 9.3, respectively. Furthermore, the amounts of hydrogen peroxide and ozone in aqueous were quantitatively measured to evaluate their roles during antibiotic removal, and the main function of hydroxyl radicals was demonstrated by the radicals scavenger test. At last, the analyses of UV-Vis spectra and HPLC-MS were employed to study the OTC elimination mechanism, and the possible decomposition pathway was proposed based on the speculated intermediates.

Chemical adsorption of oxytetracycline from aqueous solution by modified molecular sieves.

The removal of oxytetracycline (OTC) from aqueous solution on modified molecular sieve via adsorption was investigated in the present work. The copper(II) modified molecular sieve had the much higher adsorbed amount than unmodified one. The bigger pore, the more adsorption sites benefitted for the adsorbed amount of OTC. The exchanged amount of copper(II) and the acid-base property of solution were important factors influencing the removal efficiency. The adsorption kinetics, the adsorption isotherm, the adsorption thermodynamics and the proposed adsorption mechanism were studied. The analysis of adsorption isotherm indicated it is a monolayer adsorption. The fitting with adsorption kinetics, pseudo-second-order model, deduced chemical adsorption is the main rate controlling step. And the new formation of Cu-O chemical bond and the changes at bands of N-H vibration and C-N vibration by Fourier transform infrared spectrometer further confirmed the proposal adsorption mechanism was the chemical complexation of copper(II) in modified 13X with NH2 group of OTC. As the real exchanged amount of copper(II) was 149.07 mg·g-1 and the solution pH 7.0, the adsorption capacity of modified 13X for OTC reached the maximum of 2,396 mg·g-1 (with the initial concentration of 1,000 mg·L-1).

Oxytetracycline recovery from aqueous media using computationally designed molecularly imprinted polymers.

Polymers for recovery/removal of the antimicrobial agent oxytetracycline (OTC) from aqueous media were developed with use of computational design and molecular imprinting. 2-Hydroxyethyl methacrylate, 2-acrylamide-2-methylpropane sulfonic acid (AMPS), and mixtures of the two were chosen according to their predicted affinity for OTC and evaluated as functional monomers in molecularly imprinted polymers and nonimprinted polymers. Two levels of AMPS were tested. After bulk polymerization, the polymers were crushed into particles (200-1000 µm). Pressurized liquid extraction was implemented for template removal with a low amount of methanol (less than 20 mL in each extraction) and a few extractions (12-18 for each polymer) in a short period (20 min per extraction). Particle size distribution, microporous structure, and capacity to rebind OTC from aqueous media were evaluated. Adsorption isotherms obtained from OTC solutions (30-110 mg L(-1)) revealed that the polymers prepared with AMPS had the highest affinity for OTC. The uptake capacity depended on the ionic strength as follows: purified water > saline solution (0.9 % NaCl) > seawater (3.5 % NaCl). Polymer particles containing AMPS as a functional monomer showed a remarkable ability to clean water contaminated with OTC. The usefulness of the stationary phase developed for molecularly imprinted solid-phase extraction was also demonstrated. Graphical Abstract Selection of functional monomers by molecular modeling renders polymer networks suitable for removal of pollutants from contaminated aqueous environments, under either dynamic or static conditions.

Biodegradation of three tetracyclines in swine wastewater.

Tetracyclines (TCs), including tetracycline (TC), oxytetracycline (OTC), and chlortetracycline (CTC), are amongst the most common antibiotics used in animal husbandry. Residual amounts of these antibiotics in the environment are a concern because they contribute to selection of resistant bacteria. In this study, we investigated the biodegradation of three TCs in swine wastewater. In batch experiments, OTC and CTC were completely degraded at d 18 and 20, respectively, but TC was remained at 7.1% after 20 d incubation. The degradation rates of TCs in the wastewater were in the order of OTC > CTC > TC. Degradation of the TCs was enhanced by the addition of enzyme extract from spent mushroom compost (SMC) of Pleurotus eryngii. The degradation rates were higher with the addition of extract-containing microcapsules than suspended enzyme extract in swine wastewater. In the bioreactor experiment, the addition of extract-containing microcapsules enhanced the removal rates of the three TCs, and adding TCs twice maintained enzyme activity in the swine wastewater. Of the microorganism strains isolated from the wastewater samples, strain HL2 (identified as Xanthobacter flavus) showed the best degrading ability.

Removal of oxytetracycline from pharmaceutical wastewater using kappa carrageenan hydrogel.

This study investigated the adsorption of Oxytetracycline (OTC) from pharmaceutical wastewater using a kappa carrageenan based hydrogel (KPB). The aim of the present study was to explore the potential of KPB for long-term pharmaceutical wastewater treatment. A sustainable adsorbent was developed to address oxytetracycline (OTC) contamination. The hydrogel's structural and adsorption characteristics were examined using various techniques like Scanning Electron Microscope (SEM), Fourier Transform Infrared (FTIR), X-ray powder diffraction (XRD), and kinetic models. The results revealed considerable changes in the vibrational modes and adsorption bands of the hydrogel, suggesting the effective functionalization of Bentonite nano-clay. Kappa carrageenan based hydrogel achieved the maximum removal (98.5%) of OTC at concerntration of 40 mg/L, pH 8, cotact time of 140 min and adsorbent dose of 0.1 g (KPB-3). Adsorption of OTC increased up to 99% with increasing initial concentrations. The study achieved 95% adsorption capacity for OTC using a KPB film at a concentration of 20 mg/L and a 0.1 g adsorbent dose within 60 min. It also revealed that chemisorptions processes outperform physical adsorption. The Pseudo-Second-Order model, which emphasized the importance of chemical adsorption in the removal process, is better suited to represent the adsorption behavior. Excellent matches were found that R2 = 0.99 for KPB-3, R2 = 0.984 for KPB-2 and R2 = 0.989 for KPB-1 indicated strong chemical bonding interactions. Statisctical analysis (ANOVA) was performed using SPSS (version 25) and it was found that pH and concentration had significant influence on OTC adsorption by the hydrogel, with p-values less than 0.05. The study identified that a Kappa carrageenan-based hydrogel with bentonite nano-clay and polyvinyl alcohol (PVA) can efficiently remove OTC from pharmaceutical effluent, with a p-value of 0.054, but weak positive linear associations with pH, temperature, and contact time. This research contributed to sustainable wastewater treatment and environmental engineering.

Adsorptive removal of trace oxytetracycline from water by acid-modified zeolite: influencing factors.

Because of the wide use of antibiotics in the livestock industry, trace tetracycline antibiotics are frequently detected in swine wastewater and water bodies near pig farms. Based on natural zeolite, modified zeolite was synthesized by treatment with nitric acid. As one kind of typical tetracyclines, oxytetracycline (OTC) was chosen as the target adsorbate. Removal of trace OTC by modified zeolite and the effects of several main water matrices on OTC adsorption were studied in detail. OTC removal efficiency by acid-modified zeolite was about 90%, compared to less than 20% by natural zeolite. In general, in acidic conditions the removal efficiency of OTC by modified zeolite was about 90%, which was much higher than 20-35% in alkaline conditions. An increase in ionic strength from 0.01 to 1.0 M led to a decrease in adsorption efficiency from 90 to 27%. The presence of 10.0 mg L(-1) dissolved humic acid accelerated sorption of OTC on modified zeolite, while 100.0 mg L(-1) humic acid resulted in the opposite effect. An increase in temperature contributed to enhancing the adsorption efficiency.

Heterologous expression and manipulation of three tetracycline biosynthetic pathways.

A very accommodating host: Three tetracycline biosynthetic pathways were overexpressed and manipulated in the heterologous host Streptomyces lividans K4-114. Through the inactivation of various genes and characterization of the resulting biosynthetic intermediates, new tetracycline-modifying enzymes were identified (see scheme).

Synergistic effect of Cu-nanoparticles and ß-cyclodextrin functionalized reduced graphene oxide nanocomposite on the adsorptive remediation of tetracycline antibiotics.

Pollution by tetracyclines antibiotics has a great potential risk on human and animal health even at trace levels. Copper nanoparticles immobilized-ß-cyclodextrin functionalized reduced graphene oxide (Cu/ß-CD/rGO) were successfully prepared as an efficient extractor of tetracycline (TC), oxytetracycline (OTC) and doxycycline (DC) antibiotics from different environmental water samples. Tetracyclines (TCs) are strongly deposited in the matrix of Cu/ß-CD/rGO nanocomposite via surface complexation with the Cu-nanoparticles besides the formation of inclusion complexes with ß-cyclodextrin and π-π interaction of reduced graphene oxide. The novel nanocomposite was characterized by HRSEM, TEM, TGA, FT-IR, XPS, and XRD. The optimization of variables such as the pH, contact time, ionic strength and TC concentration were successfully analyzed. The maximum adsorption capacity (qm) of Cu/ß-CD/rGO calculated from the Langmuir isotherm was 403.2 mg.g-1 for TC, 476.2 mg.g-1 for OTC and 434.8 mg.g-1 for DC at 298 K. The removal efficiency was decreased by 3.7% after five adsorption-desorption cycles. The Cu/ß-CD/rGO nanocomposite was applied for removing TCs from tap water and the Nile River water samples. The novel nanocomposite demonstrated fast and highly efficient removing performance for different TCs with low levels and large sample volume.

Tetracycline residues in royal jelly and honey by liquid chromatography tandem mass spectrometry: validation study according to Commission Decision 2002/657/EC.

A specific, sensitive and robust liquid chromatography tandem mass spectrometry method for determining oxytetracycline, tetracycline, chlortetracycline and doxycycline in royal jelly and honey samples is presented. Extraction of drug residues was performed by ammonium acetate buffer as extractant followed by a clean-up with metal chelate affinity chromatography and solid-phase extraction. Tetracycline analysis was performed using liquid chromatography-electrospray ionisation-tandem mass spectrometry. The presented method is the first validated for royal jelly and in accordance with the requirements set by Commission Decision 2002/657/EC. Recoveries of the methods, calculated spiking the samples at 5.0, 10.0, 20.0 and 30.0 µg kg(-1), were 79% to 90% for honey and 77% to 90% for royal jelly. The intra-day precision (RSD) ranged between 8.1% and 15.0% for honey and from 9.1% to 16.3% for royal jelly, while inter-day precision values were from 10.2% to 17.6% and from 10.6% to 18.4% respectively for honey and royal jelly. Linearity for the four analytes was calculated from 5.0 to 50.0 µg kg(-1). The decision limits (CCα) ranged from 6.2 to 6.4 µg kg(-1) and from 6.1 to 6.5 µg kg(-1) for honey and royal jelly, respectively. Detection capabilities values (CCß) ranged between 7.2 and 7.7 µg kg(-1) and from 7.3 to 7.9 µg kg(-1) respectively for honey and royal jelly. The developed method is currently in use for confirmation of the official control analysis of honey and royal jelly samples.

Development of an efficient extraction method for oxytetracycline in animal manure for high performance liquid chromatography analysis.

Oxytetracycline (2-(amino-hydroxy-methylidene)-4-dimethylamino-5,6,10,11,12a-pentahydroxy-6-methyl-4,4a,5,5a-tetrahydrotetracene-1,3,12-trione) is a major member of the tetracycline antibiotics family of which are widely administered to animals in concentrated animal feeding operations for purposes of therapeutical treatment and health protection. With the disposal of animal manure as fertilizer into agricultural land, tetracyclines enter the environment. However, tetracyclines chelate with multivalent cations and proteins, resulting in low extraction efficiencies from animal manure for tetracycline residue analysis. In this study an efficient extraction method for oxytetracycline from steer manure using methanol/water solution amended with chelating organic acid was developed for the analysis of high performance liquid chromatography. The effect of species and amount of amendment acids, shaking time, methanol/water ratio, manure weight, and repeated times of extraction was investigated. It was optimized to amend 2.5 g citric acid and 1.1 g oxalic acid with 10.0 g manure sample in a 50-ml centrifuge tube and extract with 15 ml methanol/water (9:1 in volume) by vigorously shaking for 30 min in a reciprocating shaker. After centrifugation at 11,000 rpm, supernatant is collected. Sample was extracted for a total of 3 times. The developed extraction method was further applied to extract oxytetracycline from fresh and aged cow manure, swine and poultry manure, and soil. Satisfactory recoveries ranging from (84.1 +/- 2.4) % to (102.0 +/- 3.1) % were obtained, demonstrating that the optimized extraction method is robust for oxytetracycline from different manure sample matrixes.

Halloysite nanoclay supported adsorptive removal of oxytetracycline antibiotic from aqueous media.

Halloysite nanoclay was utilized to retain aqueous oxytetracycline (OTC) which is extensively used in the veterinary industry. The micro-structure and functionality of the nanoclay were characterized through spectroscopic techniques before and after adsorption. The OTC removal experiments were performed at different pH conditions (pH 3.0-9.0), ionic strengths (0.001, 0.01, 0.1 M NaNO3) and contact time (up to 32 h) at an initial 25 mg/L OTC concentration with 1.0 g/L halloysite. Oxytetracycline adsorption was pH dependent, and the best pH was observed in the range of pH 3.5-5.5 at a 0.001 M ionic strength. At pH 3.5, the maximum OTC adsorption amount was 21 mg/g which translated to 68% removal of the initial OTC loading. Positively charged inner lumen and negatively charged outer lumen of the tubular halloysite structure led to form inner-sphere complexes with the anionic and cationic forms of OTC, respectively. A rapid adsorption of OTC was observed in the kinetic study where 62% OTC was adsorbed in 90 min.. Pseudo-second order equation obeyed by the kinetic data indicated that the adsorption was governed by chemisorption, whereas Hill isotherm equation was the most fitted with a maximum adsorption capacity of 52.4 mg/g indicating a cooperative adsorption phenomenon.

Simultaneous determination of olaquindox, oxytetracycline and chlorotetracycline in feeds by high performance liquid chromatography with ultraviolet and fluorescence detection adopting online synchronous derivation and separation.

Olaquindox, oxytetracycline and chlorotetracycline were widely used in feed as antibiotics and growth promoter to improve feed conversion efficiency and increase the rate of weight gain for animals. However, the use of these antibiotics in feed was gradually prohibited because of concerns about contamination and resistance in animals. A quantitative and confirmatory method for determining the presence of olaquindox, oxytetracycline and chlorotetracycline in feed by high performance liquid chromatography equipped with ultraviolet detector in series with fluorescence detector (HPLC-UVD-FLD) was developed, optimized, and validated in three different matrices (compound, concentrated and premix feed). The analytes extraction was performed with a mixture of acetonitrile and 0.1 mol/L ethylenediamine tetraacetic acid disodium-Mcllvaine buffer (1:4, v/v) by one step sample preparation procedure. The validated method presented a broad linear range and good linearity with weighted least square method. The decision limit of the analytes ranged from 0.61 to 0.77 mg/kg for olaquindox, 0.90 to 1.2 mg/kg for oxytetracycline and 1.3 to 2.0 mg/kg for chlorotetracycline. The average recovery values found in intermediate precision conditions were ranged from 88.0 to 99.7% for olaquindox with RSD lower than 11.1%, from 84.4 to 99.0% for oxytetracycline with RSD lower than 9.6%, from 83.8 to 97.5% for chlorotetracycline with RSD lower than 10.0%. By Youden test and bottom-up method, the method was proved to be sufficiently robust and had a small uncertainty for different concentration levels. The developed method was successfully utilized for commercial feed samples to monitor complex cross contamination and residue conditions. Online synchronous derivation and separation using ultraviolet detector in series with fluorescence detector can effectively prevent false positive of chlorotetracycline in feed caused by vegetable meal. Since olaquindox, oxytetracycline and chlorotetracycline are widely used in feed, the developed method provide an important and analytical tool for the simultaneous identification and quantification of them in feed to monitor its risk of cross contamination and excessive content.

Construction of Ce-MOF@COF hybrid nanostructure: Label-free aptasensor for the ultrasensitive detection of oxytetracycline residues in aqueous solution environments.

Porous organic framework (COF) nanomaterials have drawn increasing attention and showed promising potential in the applications of various fields. Nevertheless, its applications in biosensing or biomedical fields are still in the early stage. In this work, we designed and synthesized a series of nanohybrids of COF and Ce-based metal organic framework (Ce-MOF) for the first time as label-free bioplatforms for a sensitive electrochemical aptasensor to detect oxytetracycline (OTC). A novel kinds of Ce-MOF@COF hybrids were prepared by adding different dosages of COF, into the preparation system of Ce-MOF, for which COF was synthesized using melamine and cyanutic acidmonomers through polycondensation (represented by MCA). Basic characterizations revealed that Ce-MOF@MCA nanohybrids not only remained their orignal crystal and chemical structure and features, such as different Ce species containing in Ce-MOF (Ce3+ and Ce4+), various functional amino-groups of MCA, and individual frameworks, but also showed a large specific surface area and interpenetrated morphologies. As a result, the Ce-MOF@MCA hybrid with high content of MCA exhibited high bioaffinity toward the OTC-targeted aptamer, further leading to the incremental detection effect for OTC detection. Among different hybrid-based aptasensors, the Ce-MOF@MCA-based one with an MCA dosage of 500 mg exhibited the lowest limit of detection at 17.4 fg mL-1 within a wider linearity of the OTC concentration within 0.1-0.5 ng mL-1. Additionally, the fabricated aptasensor displayed excellent analytical performance with great reproducibility, high selectivity and stability, and acceptable applicability for detecting OTC in various aqueous solutions, including milk, wastewater, and urine samples. This new Ce-MOF@MCA hybrid will become an excellent aptasensors platform for detecting various analytes, such as antibiotics, heavy metal ions, or cancer markers, and it have shown the promissing application potentials in the fields of biomedicine, food safety and environmental monitoring.

Contrastive removal of oxytetracycline and chlortetracycline from aqueous solution on Al-MOF/GO granules.

The presence of tetracycline antibiotics (TCS) in the water and wastewater has raised growing concern due to its potential environmental impacts; thus, their removal is of high importance. In this study, a novel aluminum-based MOF/graphite oxide (Al-MOF/GO) granule was prepared as an adsorbent for the removal of TCS including oxytetracycline (OTC) and chlortetracycline (CTC). The adsorbent was characterized via XRD, FTIR, BET, SEM, and XPS methods. The granules exhibited similar crystal structure and some new mesopores appearing compared to the parent Al-MOF/GO powder. In addition, the adsorption behavior of OTC and CTC on samples was explored as a function of initial concentration, contact time, pH, and ionic strength by means of batch experiments. The adsorption capacity reached to 224.60 and 240.13 mg·L-1 for OTC and CTC, at C0 = 60 mg·L-1 as well as ambient temperature respectively. Moreover, the adsorption process of OTC and CTC on Al-MOF/GO samples can be better delineated by pseudo-second-order kinetics and Freundlich isotherm models. Besides, the adsorption mechanism over Al-MOF/GO granules was proposed, which could be ascribed to π-π interaction, cation-π bonding, and hydrogen bond. Finally, the great water stability, separation performance, and regeneration efficiency of these novel granules indicated their potential application in the OTC and CTC removals from aqueous solution.

Reversed-phase high-performance liquid chromatography coupled to ultraviolet and electrospray time-of-flight mass spectrometry on-line detection for the separation of eight tetracyclines in honey samples.

Eight antibiotics, chlortetracycline, demeclocycline, doxycycline, methacycline, minocycline, oxytetracycline, tetracycline and rolitetracycline, were separated and quantified in Spanish honey extracts of different floral origin using a commercial RP-C18 HPLC column and two different on-line detectors (diode array and electrospray time-of-flight mass spectrometry (ESI-TOF-MS) systems). Operating a linear gradient at a flow of 2 ml min(-1) the HPLC separation of the eight antibiotics was obtained within 10 min with good peak symmetry and an acceptable resolution (2.1) for the critical band pair rolitetracycline and oxytetracycline. Values of the numbers of theoretical plates (N) were comprised between 2328 and 19448 while the limits of detection in honey were within 0.02-1.03 microg kg(-1) in the case of UV detection and 0.05-0.76 microg kg(-1) for ESI-TOF-MS detection (operating in negative mode). A recovery study was carried out by preparing some quality control samples at four levels of concentration (10, 25, 50 and 100 microg kg(-1)) and percentages between 72% and 98% were attained.

[Oxytetracycline Wastewater Treatment in Microbial Fuel Cells and the Analysis of Microbial Communities].

Oxytetracycline (OTC) as an important broad spectrum antibiotic has been widely used in animal husbandry. However, the abuse of OTC not only has a direct adverse impact on ecosystems, but also exacerbates the appearance of antibiotic resistance bacteria and antibiotic resistance genes. OTC-containing wastewater was treated using microbial fuel cells (MFCs), and the removal efficiency of OTC in MFCs in different operation periods was investigated. Result showed that the removal efficiency of 10 mg·L-1 OTC by MFCs within 132 h was up to 99.0% after 150 d of operation. The bacterial communities in raw pig mature and anodic biofilms were studied by high-throughput sequencing. This showed that Firmicutes were both dominant on phylum a level; However, compared to the raw pig mature, the abundance of Proteobacteria greatly increased from 2.84% to 8.92%-22.75% in the anodic biofilm. In addition, the abundance of Eubacterium spp. in the anodic biofilm increased obviously from nearly 0.00% to 20.49%-49.00%. It has been reported that some Eubacterium spp. were able to biodegrade oxygen heterocyclic aromatic compounds contained in OTC. Therefore, Eubacterium spp. is suggested as potential functional species in the biodegradation of OTC and/or its metabolites. This work proves the feasibility and effectiveness of removing OTC-containing wastewater by using MFCs.

Removal mechanism of di-n-butyl phthalate and oxytetracycline from aqueous solutions by nano-manganese dioxide modified biochar.

In this work, nano-manganese dioxide (nMnO2)-modified biochar (BC) was synthesized in order to improve BC's adsorption capacity for di-n-butyl phthalate (DBP) and oxytetracycline (OTC). The results showed that nMnO2 on the BC surface exhibited a poor crystallinity and oxidation state (Mn (IV)). Sorption experiments showed that, compared to BC, DBP sorption capacity of nMnO2-BC (1:20) and OTC sorption capacity of nMnO2-BC (1:10) were 0.0364 and 0.0867 mmol/g, respectively, which are significantly higher than that of BC (0.0141 and 0.0151 mmol/g). Kinetics and isotherm experiments indicated that physical adsorption and chemical interactions have both exerted their impacts on the adsorption process. Further X-ray photoelectron spectroscopy (XPS) analysis showed that part of the Mn (IV) in nMnO2-BC was reduced to Mn (III) and Mn (II) after DBP or OTC adsorption. Therefore, we suggest the nMnO2 also acted as an oxidizer on modified BC, which may accelerate the degradation of DBP and OTC.