Immunotechniques for the Group Determination of Macrolide Antibiotics Traces in the Environment Using a Volume-Mediated Sensitivity Enhancement Strategy.

Macrolide antibiotics, which are effective antimicrobial agents, are intensively used in human and veterinary medicine, as well as in agriculture. Consequently, they are found all over the world as environmental pollutants, causing harm to sensitive ecological communities and provoking a selection of resistant forms. A novel azithromycin derivative, which was used as hapten conjugate, ensured the group immunorecognition of six major macrolide representatives (105-41%), namely erythromycin, erythromycin ethylsuccinate, clarithromycin, roxithromycin, azithromycin, and dirithromycin in a competitive immunoassay based on anti-clarithromycin antibodies. The heterologous hapten-based ELISA format resulted in a 5-fold increase in sensitivity, with an IC50 value of 0.04 ng/mL for erythromycin. In this study, we proposed an underexploited strategy in an immunoassay field to significantly improve the detectability of analytes in environmental samples. Unlike most approaches, it does not require special enhancers/amplifiers or additional concentration/extraction procedures; instead, it involves analyzing a larger volume of test samples. A gradual volume increase in the samples (from 0.025 to 10 mL) analyzed using a direct competitive ELISA, immunobeads, and immunofiltration assay formats based on the same reagents resulted in a significant improvement (more than 50-fold) in assay sensitivity and detection limit up to 5 and 1 pg/mL, respectively. The suitability of the test for detecting the macrolide contamination of natural water was confirmed by the recovery of macrolides from spiked blank samples (71.7-141.3%). During 2022-2023, a series of natural water samples from Lake Onega and its influents near Petrozavodsk were analyzed, using both the developed immunoassay and HPLC-MS/MS. The results revealed no contamination of macrolide antibiotic.

LC-MS/MS Determination of Antibiotic Residues in Distillers Grains: Method Modification.

BACKGROUND: Antibiotics are used in ethanol production to discourage undesirable bacteria growth. To determine if antibiotic residues remain in the distillers grain (DG) byproduct, which is used as an animal food ingredient, the U.S. Food and Drug Administration/Center for Veterinary Medicine previously developed an LC-MS/MS method to detect residues of erythromycin A, penicillin G, virginiamycin M1, and virginiamycin S1 in DG to enable regulatory decision-making. OBJECTIVE: Erythromycin and penicillin G were quantitated using the stable isotope dilution technique with their isotopically labeled compounds, which are considered optimal internal standards (ISTDs) for quantitative mass spectrometry. With the commercial availability of virginiamycin M1-d2 since then, the objectives of this study were to evaluate the feasibility of its use as it is only doubly deuterated, and to incorporate it in the method to enhance method performance. METHOD: Antibiotic residues were solvent-extracted from DG; the extract was cleaned up by a hexane wash and solid phase extraction (SPE) and analyzed by LC-MS/MS. RESULTS: We established suitability of virginiamycin M1-d2 as an ISTD and incorporated it in the method. For all analytes, accuracy and precision ranged 90 to 102% and 3.8 to 6.8, respectively. CONCLUSIONS: We modified a previously developed LC-MS/MS method that uses virginiamycin M1-d2 as an ISTD to support surveillance studies to determine several drugs in DG. HIGHLIGHTS: Virginiamycin M1-d2 was successfully incorporated into the method for better virginiamycin M1 quantitation. This addition also allowed calibration curves for all analytes to be constructed in solvent, thereby simplifying the method.

Simulation and risk assessment of typical antibiotics in the multi-media environment of the Yangtze River Estuary under tidal effect.

Frequent human activities in estuary areas lead to the release of a large number of antibiotics, which poses a great threat to human health. However, there are very limited studies about the influence of the special natural phenomena on the occurrence and migration of antibiotics in the environment. In this study, we simulated the migration and transformation of six typical antibiotics, including oxytetracycline (OTC), tetracycline (TC), norfloxacin (NOR), ofloxacin (OFX), erythromycin (ETM), and amoxicillin (AMOX), in the environmental media from 2011 to 2019 in the Yangtze River Estuary, by using the level III multi-media fugacity model combined with the factor of tides. The simulation results showed that the most antibiotics mainly existed in soil and sediment while erythromycin were found mainly in water. The concentrations of antibiotics in air, freshwater, seawater, groundwater, sediment, and soil were 10-23-10-25, 0.1-12 ng/L, 0.02-7 ng/L, 0.02-16 ng/L, 0.1-13 ng/g, and 0.1-15 ng/g respectively. Sensitivity analysis showed that the degradation rate (Km) and the soil-to-water runoff coefficient (Kl) were important model parameters, indicating that hydrodynamic conditions had a significant impact on the migration of antibiotics in various environmental phases in estuarine areas. Tide can enhance the exchange between water bodies and cause the transformation of the antibiotics from freshwater to seawater and groundwater, which improved the accuracy of the model, especially the seawater and soil phase. Risk assessments showed that amoxicillin, erythromycin, ofloxacin, and norfloxacin posed a threat to the estuarine environment, but the current source of drinking water did not affect human health. Our findings suggested that, when one would like to exam the occurrence and migration of antibiotics in environment, more consideration should be given to the natural phenomena, in addition to human activities and the nature of antibiotics.

Electrochemical oxidation of amoxicillin, ciprofloxacin and erythromycin in water: Effect of experimental factors on COD removal.

An alternative method of electrochemical oxidation was employed to degrade persistent compounds in the form of antibiotics using strong oxidizing agents such as hydroxyl ions. A 24 factorial design was employed to check the effect of four factors namely pH, current density, electrolysis time and electrolyte concentration set at their high (+) and low (-) levels on the antibiotics (amoxicillin, ciprofloxacin and erythromycin) degradation in water. The response was obtained in the form of COD (chemical oxygen demand) removal. A prediction model was developed to predict the values of COD removal. Later the main effect, contribution and interactions were studied with Design Expert Software 7.0. About 89.5% COD removal was obtained when pH and time were set at their high level and the other two factors at their low level. It was determined that the pH when set at high level (pH 9) had the most effect (24.68) and contribution (43.6) in the degradation process and hence the removal of COD. This technology of electrochemical oxidation can be employed in industries to efficiently remove pharmaceuticals, paints, dyes and other organic compounds.

Preparation and characterization of molecularly imprinted polymers based on ß-cyclodextrin-stabilized Pickering emulsion polymerization for selective recognition of erythromycin from river water and milk.

Molecularly imprinted polymers were prepared via ß-cyclodextrin-stabilized oil-in-water Pickering emulsion polymerization for selective recognition and adsorption of erythromycin. The synthesized molecularly imprinted polymers were spherical in shape, with diameters ranging from 20 to 40 µm. The molecularly imprinted polymers showed high adsorption capacity (87.08 mg/g) and adsorption isotherm data fitted well with Langmuir model. Adsorption kinetics study demonstrated that the molecularly imprinted polymers acted in a fast adsorption kinetic pattern and the adsorption features of molecularly imprinted polymers followed a pseudo-first-order model. Adsorption selectivity analysis revealed that molecularly imprinted polymers had a much better specificity for erythromycin than that for spiramycin or amoxicillin, and the relative selectivity coefficient values on the bases of spiramycin and amoxicillin were 3.97 and 3.86, respectively. The Molecularly imprinted polymers also showed a satisfactory reusability after four times of regeneration. In addition, molecularly imprinted polymers exhibited good adsorption capacities for erythromycin under complicated environment, that is, river water and milk. These results proved that the as-prepared molecularly imprinted polymers is a potent absorbent for selective recognition of erythromycin, and therefore it may be a promising candidate for practical applications, such as wastewater treatment and detection of erythromycin residues in food.

Impedimetric electronic tongue based on molybdenum disulfide and graphene oxide for monitoring antibiotics in liquid media.

Antibiotics are considered emerging pollutants which indiscriminate use has led to the development of antibiotic-resistant bacteria, while their improper disposal has caused adverse effects to the environment and human health. Thus, the development of devices or techniques capable of detecting antibiotics with high sensitivity, low detection limits, and reasonable cost becomes of prime importance. In this work, an electronic tongue (e-tongue) based on molybdenum disulfide (MoS2) and graphene oxide (GO) was developed and employed to detect four distinct antibiotics, namely cloxacillin benzathine, erythromycin, streptomycin sulfate, and tetracycline hydrochloride. The five sensing units of the e-tongue were obtained using the drop-casting method to modify gold interdigitated electrodes with MoS2 and GO. Using Principal Component Analysis to process the experimental data allowed the e-tongue to recognize samples contaminated with distinct antibiotics at varied concentrations from 0.5 to 5.0 nmol L-1. Analyses with real samples were also performed using river water and human urine and the electronic tongue was able to differentiate the samples at a nanomolar level. The proposed system represents a sensitive and low-cost alternative for antibiotic analyses in different liquid media.

Molecularly imprinted polymer-based sensor for electrochemical detection of erythromycin.

The increasing global reports on the occurrence of macrolide antibiotics resistance, especially erythromycin (Ery) resistant strains, suggests the possible presence of these antibiotics in the environment hence, their inclusion in the EU watchlist of water pollutants. Consequently, there is an urgent need for the development of portable and cost effective analytical sensing devices for their monitoring in water. The combination of molecularly imprinted polymer (MIP) as a sensing element with a portable electrochemical transducer such as screen printed electrode (SPE) may offer a valuable approach for the desired routine environmental monitoring. This work demonstrates the preparation of an electrochemical MIP-based sensor for Ery detection in aqueous media. Ery-selective MIP, Ery-MIP was generated directly on SPE, Ery-MIP/SPE via electrochemical polymerization of m-phenylenediamine (mPD). The optimization of sensor performance was achieved with special attention given to the selection of functional monomer, template removal, polymer thickness and incubation time. Ery-MIP/SPE sensor demonstrated the ability to discriminate target analyte against very close analogues i.e clarithromycin and azithromycin in both PBS and tap water. In addition, Ery-MIP/SPE could detect Ery down to low limits (LOD = 0.1 nM and LOQ = 0.4 nM) and exhibited good recovery in tap water. The presented analytical approach could be potentially suited and/or further developed for adequate monitoring of Ery as well as other macrolides in environmental water.

Profiling and characterization of odorous volatile compounds from the industrial fermentation of erythromycin.

Complaints caused by odors from the fermentation production of pharmaceuticals are common in China. The elimination of odor remains a challenge for the pharmaceutical industry to meet the increasingly strict environment regulations. Erythromycin is a representative antibiotic produced by microbial fermentation. The fermentation exhaust gas of erythromycin fermentation has an unpleasant odor, but the composition of the key odorants has not been identified. The major odorants from the fermentation production of erythromycin API were analyzed by electronic nose, olfactory measurements, gas chromatography-coupled ion mobility spectrometry (GC-IMS) and gas chromatography-mass spectrometry (GC-MS) analysis. Two compounds, 2-methylisoborneol (2-MIB) and geosmin, were identified as the major odorants of erythromycin fermentation. These had not been detected before using only GC-MS analysis of exhaust gas. Aldehydes, including hexanal, octanal, decanal, and benzaldehyde, also contribute to the odor. The composition analysis of odorants using the fermentation broth headspace was more efficient and reliable, considering the significant dilution effect of exhaust gas. The concentration of 2-MIB and geosmin in the fermentation broth greatly exceeded their odor thresholds. The production of major odorants started in the early fermentation stage and became significant in the middle stage (30-70 h). Due to the extremely low odor thresholds of 2-MIB and geosmin, advanced purification may require deodorization of erythromycin fermentation exhaust gas.

Occurrence and distribution of pharmaceutical compounds in the Danshuei River Estuary and the Northern Taiwan Strait.

Ten pharmaceutically active compounds (PhACs) were determined in northern Taiwan estuarine waters and Taiwan Strait (TS) seawater. The ecological risk of these PhACs was assessed using risk quotient (RQ), which is the ratio of the measured maximum concentration to the predicted no-effect concentration. Six PhACs were detected within the estuarine waters. Caffeine concentration (130-718 ng l-1) was the highest among the analyzed PhACs. The distribution of PhACs in the Danshuei River Estuary generally exhibited addition behavior, except that caffeine showed conservative behavior. Carbamazepine, gemfibrozil, caffeine, and ketoprofen were detected in TS seawaters. Their concentrations follow the sequence: gemfibrozil > ketoprofen > caffeine > carbamazepine. The caffeine concentrations in TS seawaters were 2-3 orders of magnitude lower than those in Danshuei estuarine waters. With few exceptions for caffeine, erythromycin, and sulfadiazine posing low risk in some estuarine waters, most of the RQ values were <0.01, suggesting no adverse effects on aquatic organisms.

Cytotoxic comparison of macrolide antibiotics and their chlorinated disinfection byproduct mixtures.

Erythromycin (ERY), azithromycin (AZI) and telithromycin (TEL) are widely-used macrolide antibiotics that are frequently detected in various water environments, including resource water and drinking water. In the performed chlorination disinfection process, at least 10, 20 and 200 new disinfection byproducts of ERY, AZI and TEL, respectively, were observed (the mixtures of the disinfection byproducts of ERY, AZI and TEL were named ERY-M, AZI-M and TEL-M, respectively). There is limited information available regarding their comparative toxicities, and their potential health risks are still unknown. In this study, the Jurkat cell line was used to compare the toxicities of the disinfection byproduct mixtures and their precursor compounds. The cell viability results indicated that the toxicity of ERY-M may not be enhanced after disinfection by chlorination. In contrast, at the same concentrations, AZI-M and TEL-M induced more significant inhibitory effects on cell viability than their parent compounds. Additionally, the total antioxidant capacity (T-AOC) and cell cytokine release (including interleukin-2, interleukin-8 and tumor necrosis factor-α) analyses of AZI-M and TEL-M further verified these results. Our findings demonstrate that the cytotoxicity of AZI and TEL was enhanced during the chlorination disinfection process. This investigation will provide substantial new details related to the toxicity of the mixed disinfection byproducts (DBPs) of ERY, AZI and TEL generated in the chlorination disinfection process.

An automated on-line turbulent flow liquid-chromatography technology coupled to a high resolution mass spectrometer LTQ-Orbitrap for suspect screening of antibiotic transformation products during microalgae wastewater treatment.

The evaluation of wastewater treatment capabilities in terms of removal of water pollutants is crucial when assessing water mitigation issues. Not only the monitoring of target pollutants becomes a critical point, but also the transformation products (TPs) generated. Since these TPs are very often unknown compounds, their study in both wastewater and natural environment is currently recognized as a tedious task and challenging research field. In this study, a novel automated suspect screening methodology was developed for a comprehensive assessment of the TPs generated from nine antibiotics during microalgae water treatment. Three macrolides (azithromycin, erythromycin, clarithromycin), three fluoroquinolones (ofloxacin, ciprofloxacin, norfloxacin) and three additional antibiotics (trimethoprim, pipemidic acid, sulfapyridine) were selected as target pollutants. The analysis of samples was carried out by direct injection in an on-line turbulent flow liquid chromatography-high resolution mass spectrometry (TFC-LC-LTQ-Orbitrap-MS/MS) system, followed by automatic data processing for compound identification. The screening methodology allowed the identification of 40 tentative TPs from a list of software predicted intermediates created automatically. Once known and unknown TPs were identified, degradation pathways were suggested considering the different mechanisms involved on their formation (biotic and abiotic). Results reveal microalgae ability for macrolide biotransformation, but not for other antibiotics such as for fluoroquinolones. Finally, the intermediates detected were included into an in-house library and applied to the identification of tentative TPs in real toilet wastewater treated in a microalgae based photobioreactor (PBR). The overall approach allowed a comprehensive overview of the performance of microalgae water treatment in a fast and reliable manner: it represents a useful tool for the rapid screening of wide range of compounds, reducing time invested in data analysis and providing reliable structural identification.

Global review and analysis of erythromycin in the environment: Occurrence, bioaccumulation and antibiotic resistance hazards.

Environmental observations of antibiotics and other pharmaceuticals have received attention as indicators of an urbanizing global water cycle. When connections between environment and development of antibiotic resistance (ABR) are considered, it is increasingly important to understand the life cycle of antibiotics. Here we examined the global occurrence of erythromycin (ERY) in: 1. wastewater effluent, inland waters, drinking water, groundwater, and estuarine and coastal systems; 2. sewage sludge, biosolids and sediments; and 3. tissues of aquatic organisms. We then performed probabilistic environmental hazard assessments to identify probabilities of exceeding the predicted no-effect concentration (PNEC) of 1.0 µg L-1 for promoting ABR, based on previous modeling of minimum inhibitory concentrations and minimal selective concentrations of ERY, and measured levels from different geographic regions. Marked differences were observed among geographic regions and matrices. For example, more information was available for water matrices (312 publications) than solids (97 publications). ERY has primarily been studied in Asia, North America and Europe with the majority of studies performed in China, USA, Spain and the United Kingdom. In surface waters 72.4% of the Asian studies have been performed in China, while 85.4% of the observations from North America were from the USA; Spain represented 41.9% of the European surface water studies. Remarkably, results from PEHAs indicated that the likelihood of exceeding the ERY PNEC for ABR in effluents was markedly high in Asia (33.3%) followed by Europe (20%) and North America (17.8%). Unfortunately, ERY occurrence data is comparatively limited in coastal and marine systems across large geographic regions including Southwest Asia, Eastern Europe, Africa, and Central and South America. Future studies are needed to understand risks of ERY and other antibiotics to human health and the environment, particularly in developing regions where waste management systems and treatment infrastructure are being implemented slower than access to and consumption of pharmaceuticals is occurring.

Investigation of pharmaceutically active compounds in an urban receiving water: Occurrence, fate and environmental risk assessment.

Pharmaceutically active compounds (PhACs) recently have been recognized to constitute a health risk for aquatic ecosystems. The major pathways of PhACs to enter the aquatic environment are excretion and discharge of effluents through sewage treatment plants (STPs). The occurrence, bioaccumulation and risk assessment of lipophilic PhACs, including erythromycin, ketoconazole, indomethacin, diclofenac, gemfibrozil, bezafibrate, propranolol, carbamazepine, sertraline and 17α-ethinylestradiol were investigated in a river that receives effluents from STP. The results indicate that the PhACs were extensively existed in fish, sediment, suspended particulate matter (SPM), colloidal phase (5 kDa to 1 µm) and truly dissolved phase (< 5 kDa) water, with total concentration of ten PhACs (Σ10PhACs) of ND-19.6 ng/g, 7.3-11.2 ng/g, 25.3-101.5 ng/g, 10.1-27.7 ng/L and 67.0-107.6 ng/L, respectively. The Σ10PhACs for particulate and water samples collected from STP's outfall site were higher than those collected from upstream and downstream, indicating that the STP is an important PhACs source of river. However, the Σ10PhACs in sediment showed no significant statistical differences in the sampling area, and which was 3.5-9.5 times lower than those in SPM samples. The colloidal phase contributed 2.5-28.5% of erythromycin, 5.8-45.6% of ketoconazole, 8.4-32.2% of indomethacin, 7.0-21.4% of diclofenac, 11.6-36.9% of gemfibrozil, 10.2-45.9% of bezafibrate, 5.9-16.8% of propranolol, 1.9-11.1% of carbamazepine and 1.1-23.8% of sertraline in the aquatic environment. This suggests that aquatic particulates (e.g., colloids and SPM) maybe an important carrier for PhACs in the aquatic system. In general, the Σ10PhACs in the tissues of fish were in order as follows: kidney > brain > liver > gill > muscle. Based on truly dissolved concentrations of PhACs in the water, bioaccumulation factors were between 3.7 and 2727.3 in the fish tissues, sertraline exhibited bioaccumulation potential. In all the risk assessments, erythromycin could cause most harmful adverse health effects for the most sensitive algae group based on the acute and chronic data. In addition, the risk quotient values for diclofenac toward fish were higher than 1. These results indicate that the PhACs pose a potential risk to the aquatic organisms, especially for chronic risk.

Transfer of antibiotics from wastewater or animal manure to soil and edible crops.

Antibiotics are added to agricultural fields worldwide through wastewater irrigation or manure application, resulting in antibiotic contamination and elevated environmental risks to terrestrial environments and humans. Most studies focused on antibiotic detection in different matrices or were conducted in a hydroponic environment. Little is known about the transfer of antibiotics from antibiotic-contaminated irrigation wastewater and animal manure to agricultural soil and edible crops. In this study, we evaluated the transfer of five different antibiotics (tetracycline, sulfamethazine, norfloxacin, erythromycin, and chloramphenicol) to different crops under two levels of antibiotic-contaminated wastewater irrigation and animal manure fertilization. The final distribution of tetracycline (TC), norfloxacin (NOR) and chloramphenicol (CAP) in the crop tissues under these four treatments were as follows: fruit > leaf/shoot > root, while an opposite order was found for sulfamethazine (SMZ) and erythromycin (ERY): root > leaf/shoot > fruit. The growth of crops could accelerate the dissipation of antibiotics by absorption from contaminated soil. A higher accumulation of antibiotics was observed in crop tissues under the wastewater treatment than under manure treatment, which was due to the continual irrigation that increased adsorption in soil and uptake by crops. The translocation of antibiotics in crops mainly depended on their physicochemical properties (e.g. log Kow), crop species, and the concentrations of antibiotics applied to the soil. The levels of antibiotics ingested through the consumption of edible crops under the different treatments were much lower than the acceptable daily intake (ADI) levels.

An LC-MS/MS method for the determination of antibiotic residues in distillers grains.

Antibiotics are used in ethanol production to discourage the growth of bacteria that would result in lower ethanol content and a lower quality product. A survey conducted by the FDA (FY 2010 Nationwide Survey of Distillers Grains for Antibiotic Residues, 2009 [1]) revealed that the residues of these antibiotics can remain in the distillers grains (DG) by-product, which is used as an animal feed ingredient. The low levels of antibiotic residues in DG could be a public health concern, as they could lead to antimicrobial resistance. To enable the quantitative determination of these antibiotics (erythromycin, penicillin G, virginiamycin M1 and virginiamycin S1), we developed a sensitive LC-MS/MS method. The residues were extracted from distillers grains with a mixture of acetonitrile and buffer followed by acetonitrile. The combined extract was diluted with water and washed with hexane. An aliquot was cleaned up on an Oasis HLB solid phase extraction cartridge. Extracts were analyzed by LC-tandem mass spectrometry. The method was successfully validated using a variety of different matrices such as corn DG, corn & milo DG, and deoiled corn DG. Absolute recoveries of the analytes ranged from 53 to 106%. Accuracy ranged from 90 to 101% based on calibration by matrix standards. The limits of quantitation and relative standard deviation were all satisfactory to support future surveillance studies.

Transformation of erythromycin during secondary effluent soil aquifer recharging: Removal contribution and degradation path.

Erythromycin (ERY), a widely used antibiotic, has recently been detected in municipal secondary effluents and poses serious threats to human health during wastewater reusing. In this study, the removal, fate, and degradation pathway of ERY in secondary effluent during soil aquifer treatment was evaluated via laboratory-scale SAT tests. Up to a 92.9% reduction of ERY in synthetic secondary effluent was observed in 1.0m depth column system, which decreased to 64.7% when recharged with wastewater treatment plant secondary effluent. XRD-fractionation results demonstrated that the transphilic acid and hydrophobic acid fractions in secondary effluent compete for the adsorption sites of the packed soil and lead to a declined ERY removal. Moreover, aerobic biodegradation was the predominant role for ERY removal, contributing more than 60% reduction of ERY when recharged with synthetic secondary effluent. Destruction of 14-member macrocyclic lactone ring and breakdown of two cyclic sugars (l-cladinose and d-desosamine) were main removal pathways for ERY degradation, and produced six new intermediates.

Isolation and characterization of a high-efficiency erythromycin A-degrading Ochrobactrum sp. strain.

In this work, Erythromycin A(EA)- degrading bacteria was isolated from the contaminated soil obtained from a pharmaceutical factory in China. The isolate designated as strain WX-J1 was identified as Ochrobactrum sp. by sequence analysis of its 16S rDNA gene. It can grow in a medium containing EA as the sole source of carbon and its optimal growth pH and temperature were 6.5 and 32°C, respectively. Under these conditions, when the initial Erythromycin A concentration was 100mg·L-1, 97% of Erythromycin A has been degraded. HPLC-MS analyses indicated that Erythromycin A degradation produced intermediates contained the following three substances: 3-depyranosyloxy erythromycin A, 7,12-dyhydroxy-6-deoxyerythronolide B, 6-deoxyerythronolide B and propionaldehyde. Since Erythromycin A-degrading Ochrobactrum sp. strain rapidly degraded Erythromycin A, this strain might be useful for bioremediation purposes.

Environmental impact assessment of veterinary drug on fish aquaculture for food safety.

The degradation of veterinary drugs approved for use in aquaculture is very important in the evaluation of the impact of these drugs on the environment and to ensure safe food production. The purpose of this study is to provide guidance on how the interpretation of environmental fate data can be used by applicants to aid in protecting the environment and for the basis for food production, by suggesting the correct interpretation of data as part of an effective registration process. Tests were performed using a modification of the Organisation for Economic Co-operation and Development (OECD) 308 guideline using erythromycin and oxolinic acid under combinations of aerobic and anaerobic conditions, with and without sediment, in sea water and fresh water. Estimated DT50 s of erythromycin in fresh and sea water ranged from 6.8 to 37.9 days and estimated DT90 s were from 22.6 to 125.9 days. Degradation was more rapid in fresh water than in sea water with the formation of three degradation products: anhydroerythomycin A, erythromycin A enol ether, and pseudoerythomycin A enol ether. Estimated DT50 s of oxolinic acid in fresh and sea water were from 10.3 to 63.0 days and estimated DT90 s were from 34.3 to 209.4 days which suggests that oxolinic acid is more persistent in the environment than erythromycin. Copyright © 2016 John Wiley & Sons, Ltd.

Impacts of proline on the central metabolism of an industrial erythromycin-producing strain Saccharopolyspora erythraea via (13)C labeling experiments.

Saccharopolyspora erythraea E3 is an important industrial strain for erythromycin production and knowledge on its metabolism is limited. In the present work, (13)C labeling experiments were conducted to characterize the metabolism of S. erythraea E3. We found that S. erythraea E3 was difficult to grow on minimal medium with glucose as sole carbon source and the addition of proline remarkably improved the cell growth. The activity of EMP pathway was very low and ED pathway was alternatively the main glucose utilization pathway. The addition of proline resulted in remarkable changes in the fluxes of central metabolism. The fluxes in PP pathway, in TCA cycle and in ED pathway were 90% higher, 64% and 31% lower on Glc/Pro than on Glc, respectively. The maintenance energy on Glc/Pro was 58.4% lower than that on Glc. The energy charge was lower on Glc than on Glc/Pro, indicating that the cells on Glc suffered from energy burden. This study elucidates the impacts of proline on the central metabolism of S. erythraea and deepens the understanding of its metabolism.

Reduced persistence of the macrolide antibiotics erythromycin, clarithromycin and azithromycin in agricultural soil following several years of exposure in the field.

The macrolide antibiotics erythromycin, clarithromycin and azithromycin are very important in human and animal medicine, and can be entrained onto agricultural ground through application of sewage sludge or manures. In the present study, a series of replicated field plots were left untreated or received up to five annual spring applications of a mixture of three drugs to achieve a nominal concentration for each of 10 or 0.1mgkg(-1) soil; the latter an environmentally relevant concentration. Soil samples were incubated in the laboratory, and supplemented with antibiotics to establish the dissipation kinetics of erythromycin and clarithromycin using radioisotope methods, and azithromycin using HPLC-MS/MS. All three drugs were dissipated significantly more rapidly in soils with a history of field exposure to 10mgkg(-1) macrolides, and erythromycin and clarithromycin were also degraded more rapidly in field soil exposed to 0.1mgkg(-1) macrolides. Rapid mineralization of (14)C-labelled erythromycin and clarithromycin are consistent with biodegradation. Analysis of field soils revealed no carryover of parent compound from year to year. Azithromycin transformation products were detected consistent with removal of the desosamine and cladinose moieties. Overall, these results have revealed that following several years of exposure to macrolide antibiotics these are amenable to accelerated degradation. The potential accelerated degradation of these drugs in soils amended with manure and sewage sludge should be investigated as this phenomenon would attenuate environmental exposure and selection pressure for clinically relevant resistance.