Investigation of tylosin and tilmicosin residues in meat by high-performance liquid chromatography method.

In this study, the presence and level of macrolide group antibiotics (tylosin and tilmicosin) were analyzed by the High-Performance Liquid Chromatography (HPLC) method in a total of 126 raw meat samples, including 42 chicken breast and 84 beef neck, available for consumption in the Burdur province (Turkey). The method demonstrated good linearity (R2 > 0.999) over the assayed concentration range (0.10-10 µg/mL). Intra-day and inter-day recoveries were used to express the accuracy of the method at three different levels of 0.5, 1, 2.5 µg/mL. Intraday recoveries and relative standard deviation values ranged from 97.270 (0.054)% to 98.643 (0.061)%, and inter-day recoveries and relative standard deviation values ranged from 97.057 (0.070)% to 98.197(0.042)% for tylosin. Intraday recoveries and relative standard deviation values ranged from 96.360 (0.065)% to 98.153 (0.046)%, and inter-day recoveries and relative standard deviation values ranged from 96.050 (0.058)% to 97.053 (0.096)% for tilmicosin. The limit of detection (LOD) value was calculated as 0.473 µg/kg for tylosin, and 0.481 µg/kg for tilmicosin; the limit of quantification (LOQ) value was calculated as 1.561 µg/kg for tylosin, and 1.587 µg/kg for tilmicosin. In general, tylosin and tilmicosin were determined in the range of 8-256 µg/kg and 30-447 µg/kg, respectively, in chicken breast meat samples; also, they were detected in the range of 36-1209 µg/kg and 30-1102 µg/kg, respectively, in beef neck meat samples. It was also found that the residues of tylosin and tilmicosin in chicken and beef meats from the market were at a much higher level than the acceptable limits specified in the regulations. This creates serious problems in terms of the ecosystem, food technology, and public health, and causes significant economic losses.

Validation and application of an immunochromatographic test to detect four macrolides and two lincosamides in raw cow milk.

In this study, an immunochromatographic test (using the Charm QUAD2® Test) was used to screen for residual macrolides and lincosamides in raw cow's milk. The validation parameters (selectivity/specificity, detection capability (CCß), and ruggedness) were in agreement with the requirements of[EC] 2021. The selectivity of the immunochromatographic test was verified by the negative results of microbiological tests. The false-positive rate was 0%. The CCß values of the immunochromatographic test for various antibiotics in milk were as follows: erythromycin 0.02 mg/kg, spiramycin 0.1 mg/kg, tilmicosin 0.025 mg/kg, tylosin 0.05 mg/kg, lincomycin 0.15 mg/kg, and pirlimycin 0.15 mg/kg. The determined CCß values were lower than the respective maximum residue limits (MRLs; regulatory limits in Japan) for milk, except for lincomycin (equal to the MRL). The presence of antibiotic groups other than macrolides and lincosamides did not interfere with the specificity of the test. It showed no significant difference in lot-to-lot repeatability. The results obtained by the two researchers showed no significant differences. Finally, the test was applied to milk samples obtained from a tylosin-treated cow. The outcome was positive and in agreement with the results of the chemical analytical and microbiological methods. Therefore, this validated immunochromatographic test is expected to be suitable for routine analysis to ensure milk safety.

Health risk assessment of antimicrobial residues in sheep carcasses marketed in Kuwait.

A total of 450 samples comprising 150 each of muscles, livers and kidneys were collected from 150 sheep carcasses in Kuwait and tested by both Rapid Premi®Test kits for rapid detection of antimicrobial-positive samples, and the High-performance liquid chromatography (HPLC) for residual determination of amoxicillin, tetracycline, oxytetracycline, and tylosin. Premi test revealed that 82%, 64% and 100% of the muscle, liver and kidney samples examined, respectively were positive for antimicrobials. HPLC analysis revealed mean concentrations (µg/kg) of 45.26, 148.17, 103.18, and 71.80 for amoxicillin, oxytetracycline, tetracycline, and tylosin respectively in muscles; 64.43, 263.15, 177.04, and 112.94, respectively in livers; and 53.12, 368.21, 196.40, and 138.63, respectively in kidneys. Although many samples exceeded the maximum residue limit (MRL) of Codex Alimentarius, the assessment of dietary exposure to the antimicrobials tested through consumption of sheep meat and organs did not constitute any health hazards in the different age groups of Kuwaiti population.

Distribution of quinolone and macrolide resistance genes and their co-occurrence with heavy metal resistance genes in vegetable soils with long-term application of manure.

The spread of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) has become an increasingly serious global public health issue. This study investigated the distribution characteristics and influencing factors of ARB and ARGs in greenhouse vegetable soils with long-term application of manure. Five typical ARGs, four heavy metal resistance genes (MRGs), and two mobile genetic elements (MGEs) were quantified by real-time quantitative polymerase chain reaction (qPCR). The amount of ARB in manure-improved soil greatly exceeded that in control soil, and the bacterial resistance rate decreased significantly with increases in antibiotic concentrations. In addition, the resistance rate of ARB to enrofloxacin (ENR) was lower than that of tylosin (TYL). Real-time qPCR results showed that long-term application of manure enhanced the relative abundance of ARGs in vegetable soils, and the content and proportion of quinolone resistance genes were higher than those of macrolide resistance genes. Redundancy analysis (RDA) showed that qepA and qnrS significantly correlated with total and available amounts of Cu and Zn, highlighting that certain heavy metals can influence persistence of ARGs. Integrase gene intI1 correlated significantly with the relative abundance of qepA, qnrS, and ermF, suggesting that intI1 played an important role in the horizontal transfer of ARGs. Furthermore, there was a weakly but not significantly positive correlation between specific detected MRGs and ARGs and MGEs. The results of this study enhance understanding the potential for increasing ARGs in manure-applied soil, assessing ecological risk and reducing the spread of ARGs.

Immunochromatographic assays based on three kinds of nanoparticles for the rapid and highly sensitive detection of tylosin and tilmicosin in eggs.

Three kinds of immunochromatographic assays (ICAs) are proposed for the highly sensitive and rapid determination  of tylosin (TYL) and tilmicosin (TIM) in eggs based on colloidal gold (CG), latex microsphere (LM), and time-resolved fluorescent microsphere (TRFM). Three types of ICAs could tolerate the egg matrix via simple sample pretreatment and demonstrated high sensitivity for TYL and TIM with cut-off values of 6/6/3 µg/kg and 14/14/6 µg/kg, respectively. Furthermore, in a single-blind parallel study 20 egg samples were analyzed  by the three developed ICAs and confirmed by LC-MS/MS. The  results showed good consistency, and there were no false positive and false negative results in our three ICAs. Consequently, the proposed three ICAs offered rapid, highly sensitive, reliable, and selectable testing platforms for screening veterinary medicine or other small molecule contaminants.

Infrared spectroscopy combined with random forest to determine tylosin residues in powdered milk.

The contamination of milk by antibiotic residues is a worldwide health and food safety problem. There is a need to develop new methods for the rapid determination of antibiotic residues in milk. A method has been developed for determining tylosin residues directly in powdered milk using Fourier Transformed Infrared spectroscopy (FTIR). Tylosin is a broad-spectrum macrolide antibiotic. The spectra obtained were submitted to chemometric analysis to obtain a prediction model for tylosin concentration in powdered milk. Using the Boruta algorithm, the absorption bands related to the milk contamination by the antibiotic were identified. Random forest was shown to be adequate for the prediction of tylosin residues in milk at low concentrations (≤ 100 µg L-1) and the prediction model generated showed high correlation and determination coefficients (greater than 0.95). The proposed methodology proved to be efficient for the investigation of antibiotic residues in powdered milk.

Synthesis of molecularly imprinted polymers based on boronate affinity for diol-containing macrolide antibiotics with hydrophobicity-balanced and pH-responsive cavities.

In this research, in order to separate and purify diol-containing macrolide antibiotics, like tylosin, from complex biological samples, molecularly imprinted polymer (MIP) based on boronate affinity for tylosin was synthesized by using precipitation polymerization method with 4-vinylphenylboronic acid (VPBA) and dimethyl aminoethyl methacrylate (DMAEMA) as pH-responsive functional monomers, and N,N'-methylene bisacrylamide (MBAA)/ ethylene glycol dimethacrylate (EGDMA) as the co-crosslinkers that balance the hydrophobicity of the MIP. The synthesized tylosin-MIP had the advantages of high adsorption capacity (120 mg/g), fast pH-responsiveness responsible for the accessibility of imprinted cavities, and high selectivity coefficient towards tylosin versus its analogues (2.8 versus spiramycin, 7.3 versus desmycosin) in an aqueous environment. The mechanism of boronate affinity between tylosin and VPBA in the form of charged hydrogen bonding was analyzed via density functional theory (DFT). MIPs were used to successfully separate diol-containing macrolides through molecularly imprinted solid phase extraction (MISPE). The results show that MIPs prepared in this method have a good application prospect in the separation and purification of the diol-containing macrolide antibiotics.

When is it safe to eat different broiler chicken tissues after administration of doxycycline and tylosin mixture?

Residues of veterinary drugs in poultry meat have serious health effects on humans (e.g., antimicrobial resistance, carcinogenicity, and hypersensitivity), which make the control of veterinary drug residues an important parameter in ensuring consumer protection.  This work was performed to quantitatively determine two co-formulated anti-infective veterinary agents, tylosin tartrate (TYT) and doxycycline hydrochloride (DOX) in different tissues of broiler chickens (liver, muscles, and fat) using high performance liquid chromatography. The chicken was treated with the recommended dose of a binary mixture of the drugs (Tydovet). Moreover, the study aimed to estimate the withdrawal time of both drugs in chicken tissues. The analysis was done by solvent extraction and solid-phase extraction for clean-up of samples from the tissue matrix, followed by liquid chromatographic determination of the cited drugs with UV-detection. Residue decline with time was tracked, and both antibiotics were found to be more persistent in liver tissues than other tissues (muscle and fat). The effect of freezing and cooking was investigated on tissue residue levels. While freezing had little effect on the concentration of both antibiotics; cooking, as anticipated, led to a marked decline. Therefore, it is recommended to pay attention to the proper withdrawal periods before marketing to ensure the hygienic suitability of broilers edibles for safe human consumption. PRACTICAL APPLICATION: This novel study measures tylosin and doxycycline residues simultaneously in different tissues (muscle, fat, and liver) after administration of Tydovet powder to the broiler chicken. Residues in fat persisted for a longer time than in muscle in case of TYT, whereas the reverse was noticed in DOX.

FTIR spectroscopy with chemometrics for determination of tylosin residues in milk.

BACKGROUND: Contamination of milk by antibiotic residues represents risks to the health of consumers; therefore they should be monitored. The objective of this study was to propose a methodology for the determination of tylosin residues directly in fluid milk based on mid-infrared spectroscopy associated with chemometrics, using attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy associated with multilayer perceptron network (MLP) and partial least squares (PLS). RESULTS: MLP was shown to be adequate for the discrimination of milk samples contaminated with tylosin below or equal to or above the maximum residue limit (MRL), with an accuracy greater than 99%, using FTIR spectra data. PLS was shown to be appropriate for the prediction of the very low concentrations (0-100 µg L-1 ) of tylosin residues in milk using FTIR spectra data. PLS models with high correlation coefficients (R > 0.99) were generated. CONCLUSION: FTIR with chemometrics proved to be a non-destructive, efficient and low-cost method for the investigation and quantification of tylosin residues directly in fluid milk. © 2020 Society of Chemical Industry.

Development of a double immunochromatographic test system for simultaneous determination of lincomycin and tylosin antibiotics in foodstuffs.

This study is devoted to the development of a sensitive immunochromatographic analysis (ICA) for simultaneous determination of tylosin (TYL) and lincomycin (LIN) as antibiotics of the macrolide and lincosamide classes, widely used in animal husbandry and implicated in the contamination of foodstuffs. The ICA was implemented in an indirect competitive format, using antispecies antibodies conjugated with gold nanoparticles (GNPs) as a label. After the multistep optimization, the developed double ICA allowed for antibiotics detection with instrumental limits of detection/cutoff levels of 0.09/2 ng/mL and 0.008/0.8 ng/mL for TYL and LIN, respectively, within 10 min. The cross-reactivity was 40% to lincosamide clindamycin and negligible to other antibiotics tested. The test system allowed for the detection of TYL and LIN in milk, honey, and eggs. The recoveries of antibiotics from foodstuffs were 87.5-112.5%. The results demonstrate that the developed double ICA is an effective approach for the detection of other food contaminants.

Design of Novel Haptens and Development of Monoclonal Antibody-Based Immunoassays for the Simultaneous Detection of Tylosin and Tilmicosin in Milk and Water Samples.

In this work, a monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay (icELISA) was established to detect tylosin and tilmicosin in milk and water samples. A sensitive and specific monoclonal antibody was prepared by rational designed hapten, which was achieved by directly oxidizing the aldehyde group on the side chain of tylosin to the carboxyl group. Under the optimized conditions, the linear range of icELISA for tylosin and tilmicosin were 1.3 to 17.7 ng/mL and 2.0 to 47.4 ng/mL, with half-maximal inhibition concentration (IC50) values of 4.7 and 9.6 ng/mL, respectively. The cross-reactivity with other analogues of icELISA was less than 0.1%. The average recoveries of icELISA for tylosin and tilmicosin ranged from 76.4% to 109.5% in milk and water samples. Besides, the detection results of icELISA showed good correlations with HPLC-MS/MS. The proposed icELISA was satisfied for rapid and specific screening of tylosin and tilmicosin residues in milk and water samples.

Efficient reduction of antibiotic residues and associated resistance genes in tylosin antibiotic fermentation waste using hyperthermophilic composting.

Insufficient removal of antibiotics and antibiotic resistance genes (ARGs) from waste products can increase the risk of selection for antibiotic resistance in non-clinical environments. While composting is an efficient way to reduce ARGs, most conventional methods are ineffective at processing highly contaminated antibiotic fermentation waste. Here we explored the efficacy and underlying mechanisms of hyperthermophilic composting at removing tylosin antibiotic fermentation residues (TFR) and associated ARGs and mobile genetic elements (MGEs; plasmids, integrons and transposon). Hyperthermophilic composting removed 95.0% of TFR, 75.8% of ARGs and 98.5% of MGEs and this reduction mainly occurred after extended exposure to temperatures above 60 °C for at least 6 days. Based on sequencing and culture-dependent experiments, reduction in ARGs and MGEs was strongly associated with a decrease in the number of bacterial taxa that were initially associated with ARGs and MGEs. Moreover, we found 94.1% reduction in plasmid genes abundances (ISCR1 and IncQ-oriV) that significantly correlated with reduced ARGs during the composting, which suggests that plasmids were the main carriers for ARGs. We verified this using direct culturing to show that ARGs were more often found in plasmids during the early phase of composting. Together these results suggest that hyperthermophilic composting is efficient at removing ARGs and associated resistance genes from antibiotic fermentation waste by decreasing the abundance of antibiotic resistance plasmids and associated host bacteria.

Evaluation of Tilmicosin Contamination in Eggs Following Its Administration to Laying Hens and Subsequent Assessment of Dietary Risks to Chinese Consumers.

In the present study, we analyzed the tilmicosin residues in eggs as well as the dietary risks posed by the residual drug to Chinese consumers. Tilmicosin was administrated to laying hens via drinking water in doses of 100 mg/L and 300 mg/L (dose 1 and dose 2) for 5 days. Its residues distribution within egg matrices were detected using a QuEChERS method coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). The results indicated that the tilmicosin residue was the highest in the whole eggs and yolks on day 2 after medication and in the whites on day 5 of drug administration for dose 1 and dose 2 with the residues (µg/kg) being the following: 73.4 and 444.3 in the whole eggs; 152.0 and 1141.1 in the yolks; and 48.8 and 277.6 in the whites, respectively. During withdrawal phase, the tilmicosin levels declined to less than the limit of detection (LOD) after 29 days and 49 days for the whole eggs, after 24 days and 44 days for the yolks, and after 19 days and 49 days for the whites for doses 1 and 2, respectively. An assessment of the dietary risk for Chinese consumers revealed that the hazard quotients (HQ) values for tilmicosin consumption were less than 1 for dose 1 and greater than 1 for dose 2, with children (2 to 7 years) and adult males (over 65 years) being the high-risk groups. These results suggest that the possible risk associated with tilmicosin contamination in eggs should not be ignored since the accidental or occasional misuse of tilmicosin in the case of egg-laying hens may occur from time to time, particularly on small farms. PRACTICAL APPLICATION: The modified QuEChERS method was used to analyze the tilmicosin residues in egg matrices and a subsequent dietary exposure assessment for Chinese consumers was performed, which can serve as a reference for the food safety risk posed by antibiotic misuse in egg layers.

Removal of tylosin and copper from aqueous solution by biochar stabilized nano-hydroxyapatite.

Antibiotics and heavy metals are frequently detected simultaneously in water environment. Effective elimination methods for antibiotics and heavy metals pollution should deserve our attention. This study investigates the adsorption performance of biochar modified with nano-hydroxyapatite (nHAP) on tylosin (TYL) and Cu from water simultaneously. Composite adsorbents of nHAP and biomass, derived from three waste residues, which were wood-processing residues (WR), wheat straw (WS) and Chinese medicine residues (CMR), were prepared. According to the results of orthogonal experiment, the degree of influence of the three factors on TYL and Cu were the pyrolysis temperature > the proportion of nHAP and biomass > the sources of biomass, and pyrolysis temperature> the sources of biomass> the proportion of nHAP and biomass, respectively. The optimum conditions for nHAP@biochar were screened. At pH < 7.0, the adsorption quality of TYL increased with pH increased, while at pH > 7.0, the adsorption quality of TYL changed slightly. At low pH, Cu and TYL could compete for the same adsorption sites on nHAP@biochars. The adsorption amount of TYL and Cu were both increased with increasing of the temperature. Compared with Langmuir model, Freundlich model could better fit the TYL adsorption on nHAP@biochars, with Kf values of TYL 62.35 (mmol/kg) (L/mmol)n (WR1) and 4.84 (mmol/kg) (L/mmol)n (CMR1), respectively.

Tylosin A and desmycosin in honey by salting-out assisted liquid-liquid extraction and aqueous normal phase ultraperformance liquid chromatography-tandem mass spectrometry.

A simple and rapid method was developed for the determination of tylosin A and desmycosin residues in honey. Aliquots of honey samples were dissolved in a concentrated solution of sodium acetate and the target analytes were subsequently extracted with acetonitrile. The resulting organic extract was chromatographed under aqueous normal phase (ANP) LC conditions using a bare silica stationary phase with acidified solutions of ammonium formate in both water and 5:95 water: acetonitrile as the mobile phases. Tylosin A and desmycosin residues were measured using MS/MS in the multiple reaction monitoring (MRM) mode. Based on the analysis of replicate honey samples fortified at 5, 20, and 100 µg kg-1, the method was found to provide high accuracy and precision with average intraday trueness ranging from 90.2 to 111.2% and standard deviations of less than 7%. For spiked replicates fortified at the limit of quantification (1 µg kg-1), the intraday accuracies ranged from 72.0 to 102.7% for tylosin A and from 72.1 to 93.8% for desmycosin, with standard deviations all lower than 12%. Matrix effects were relatively minimal and consistent between honey samples which eliminated the need to perform any additional cleanup of the sample extracts prior to ANP-UPLC-MS/MS analysis. Graphical abstract.

Size-dependent adsorption of antibiotics onto nanoparticles in a field-scale wastewater treatment plant.

This work present aims to evaluate the effect of a conventional wastewater treatment process on the number of nanoparticles, and the role of nanoparticles as a carrier of antibiotics. A set of methods based on asymmetrical flow field flow fractionation coupled with multi-angle light scattering to separate and quantify nanoparticles in real wastewater was established. The characterization of nanoparticles was conducted by transmission electron microscopy, energy dispersive spectrometer, UV-visible spectrophotometer and three-dimensional excitation-emission matrix fluorescence spectroscopy. The adsorption of different sizes of nanoparticles separated from the real wastewater for four targeted antibiotics (sulfadiazine, ofloxacin, tylosin and tetracycline) was studied. The results show that the number of nanoparticles were increased in the wastewater treatment process and the size range between 60 and 80 nm was predominant in wastewater samples. The nanoparticles were mainly composed of O, Si, Al and Ca elements and organic components were in the size range of 0-10 nm. Targeted antibiotics were dominantly adsorbed onto nanoparticles with 60-80 nm size range at each stage. The concentrations of tetracycline adsorbed on nanoparticles were surprisingly increased in the end of the treatment process, while ofloxacin and tylosin had the completely opposite phenomenon to tetracycline. The pH and ionic strength definitely affected the aggregation of nanoparticles and interaction with the antibiotics. It is of great significance to give insights into nanoparticle-antibiotic assemblages for the effective treatment and avoiding the water risks due to nanoparticles' ubiquitous and their risks of carrying antibiotics.

Rapid untargeted screening for drug residues in animal tissues with liquid microjunction surface sampling probe mass spectrometry.

An untargeted screening method for the rapid identification of veterinary drug residues in incurred animal tissues using liquid microjunction surface sampling probe mass spectrometry (LMJSSP-MS) was developed. Current analytical methods for veterinary drug residue screening involve lengthy sample preparation, extraction, and instrumental analysis steps. This method identifies veterinary drug residues in several different incurred animal tissues more quickly than conventional analytical methods. This LMJSSP-MS method uses an ambient ionization technology called liquid microjunction surface sampling probe along with a data dependent scan function of a quadrupole orbitrap mass spectrometer. Collected product ion spectra are searched against the mzCloud™ online mass spectral database to identify veterinary drug residues found in incurred animal tissue samples. Examples of veterinary drugs identified with this method include flunixin, tilmicosin, pentobarbital, xylazine, and ketamine. Optimization of method parameters is described and discussed. The limit of identification (LOI) of this method is estimated to be approximately 1 µg g-1 for xylazine and ketamine.

A mass spectrometer-compatible liquid chromatographic method for the analysis of tylosin and its impurities using a superficially porous particle column.

In this study, an improved liquid chromatographic (LC) method for the analysis of tylosin and its impurities has been developed. A Kinetex EVO C18 column (150 × 4.6 mm, 2.6 µm) packed with superficially porous particles was used as stationary phase. Gradient elution was applied with two mobile phases (A and B) containing acetonitrile, water and 0.2 M ammonium acetate at different ratios (20:10:70 (v/v/v) for A and 60:10:30 (v/v/v) for B). This volatile mobile phase enables the method to be coupled with mass spectrometry (MS) for additional detection and characterization of tylosin impurities. Selectivity, sensitivity, linear calibration, accuracy, precision and robustness of this analytical method were assessed through method validation. In addition, impurities above 0.05% were characterized via LC-MS/MS. It is the first time that a MS compatible method for analysis of tylosin and its impurities is presented. Moreover, it shows a considerably shorter analysis time than previously published methods.

Distribution of hydrophilic and lipophilic antibacterial drugs in skim milk, cream, and casein.

This study determined the distribution of drugs to different milk fractions according to their physicochemical properties. Hydrophilic drugs tend to concentrate in skim milk, whereas lipophilic drugs tend to concentrate in cream. The concentration of a drug in casein is related to its degree of binding to milk proteins. Thus, we aimed to determine whether withdrawal time in whole milk differs from that in cream, casein, and skim milk. Amoxicillin and tylosin were selected as prototype hydrophilic and lipophilic drugs, respectively. The study was conducted in vitro and in vivo to determine whether in vitro conditions reflect the distribution of drugs in the different milk fractions in vivo. The in vivo study was conducted using a crossover design on 6 healthy Holstein dairy cattle. First, amoxicillin (i.m., single dose, 14 mg/kg) was administered to cows. Following a 1-wk washout period, tylosin (i.m., single dose, 15 mg/kg) was administered. Concentrations of amoxicillin and tylosin in milk and milk fractions were measured using HPLC-UV. In the in vitro study, 0.04 to 400 µg/g of amoxicillin and 0.05 to 50 µg/g of tylosin were spiked to drug-free milk and the concentrations in milk and milk fractions were measured. In addition, the percentage of total protein in milk and milk fractions was determined. Amoxicillin accumulated more in skim milk than in cream and casein, both in vitro (92%) and in vivo (73%, skim milk-to-whole milk ratio). The distribution of tylosin in whole and skim milk was similar to that of amoxicillin in the in vitro study, in contrast to the accumulation of tylosin in cream seen in vivo. However, the accumulation ratio of tylosin in cream was lower than expected. By either method, tylosin was less concentrated in casein than in skim milk and cream. The percentage of total protein was similar in skim milk and whole milk and higher than in cream. Thus, amoxicillin accumulates less in cream and casein, suggesting that these fractions would pose a lower risk to the consumer. Tylosin was still present at the maximum residue limit (50 µg/kg) 24 h after injection in the casein fraction and 48 h after injection in the cream fraction.

Contributions of the microbial community and environmental variables to antibiotic resistance genes during co-composting with swine manure and cotton stalks.

Understanding the main drivers that affect the spread of antibiotic resistance genes (ARGs) during the composting process is important for the removal of ARGs. In this study, three levels of tylosin (25, 50, and 75 mg kg-1 on a dry weight basis) were added to swine manure plus a control, which was composted with cotton stalks. Each treatment was repeated in triplicate and the ARG profiles were determined with different levels of tylosin. The top 35 genera and ARGs profiles were clustered together based on the composting time. Combined composting parameters (temperature, pH, NH4+-N, NO3-N, and moisture content) accounted for 78.4% of the total variation in the changes in the potential host bacteria. In addition, the selected five composting parameters and six phyla (including 25 potential host bacterial genera) explained 46.9% and 30.7% of the variation in the ARG profiles according to redundancy analysis, respectively. The variations in ARGs during the composting process were mainly affected by the dynamics of potential host bacteria rather than integrons and the selective pressure due to bio-Cu and bio-Zn.