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Chemical stability of chlortetracycline and chlortetracycline degradation products and epimers in soil interstitial water.

Tetracyclines and tetracycline degradation products and epimers end up in the environment. In order to predict the persistence of the potential dominating species of the chlortetracyclines in the environment, the chemical stability of chlortetracycline (CTC) and four major CTC degradation products and epimers (iso-CTC, 4-epi-CTC, anhydro-CTC, and 4-epi-anhydro-CTC) was studied in milliQ water and soil interstitial water (SIW) under environmentally relevant conditions (oxygen, light, pH (3-9), and temperature (6 degrees C and 20 degrees C)). The chemical stability of the compounds was evaluated by following the decrease in amount of parent compound over time. In order to compare the results obtained under the varying conditions, apparent pseudo-first-order rate constants (k(obs)) for the disappearance of the parent compound and corresponding apparent half-lives were calculated. A statistical evaluation of the data showed that the chemical stability of the chlortetracyclines was generally dependent on photolysis, temperature, and matrix. The presence or absence of oxygen did not influence on the chemical stability. The presence of calcium and magnesium ions in SIW is believed to account for the significant differences in half-lives between milliQ water and SIW, although numerous of other factors are believed to influence as well. Generally, the five compounds were more persistent at pH 3-4 than at pH above 5.

Calcium↗

Changes in the prevalence of resistant Escherichia coil in cattle receiving subcutaneously injectable oxytetracycline in addition to in-feed chlortetracycline compared with cattle receiving only in-feed chlortetracycline.

Information about the prevalence of antibiotic resistance in commensal enteric bacteria is of interest because these bacteria are potential indicators of selection pressure on enteric bacteria and represent a reservoir of resistance genes in potentially pathogenic bacteria. This study reports changes in the prevalence of resistance to antibiotics in commensal Escherichia coli from cattle receiving either subcutaneously injectable oxytetracycline in addition to in-feed chlortetracycline or only in-feed chlortetracycline. Resistance to 19 antibiotics was examined. The use of injectable oxytetracycline in addition to in-feed chlortetracycline was significantly associated (P < 0.05) with an increase in the prevalence of resistance only to chloramphenicol and sulfisoxazole.

Animals↗

Liquid chromatographic determination of epimerization of chlortetracycline residue to 4-epi-chlortetracycline residue in animal feed, using McIlvain's buffer as extractant.

A method for studying McIlvain's solution as a factor in epimerization of chlortetracycline in animal feed has been developed. McIlvain's solution (McBuffer) was used previously to extract oxytetracycline (OTC), tetracycline-HCl (TC-HCl), and chlortetracycline-HCl (CTC-HCl) from animal feed because it gave the best recoveries for all 3 tetracyclines; however, the McBuffer solution caused epimerization of CTC-HCl to 4-epi-chlortetracycline (4-epi-CTC). Recovery results were 30-40% lower for CTC than for OTC and TC from the same sample extract. The levels of concentration tested for CTC and 4-epi-CTC were 10 ppm, the detection limits for both were 3 ppb, and the calibration curves were linear between 5 and 80 ppm for both CTC-HCl and 4-epi-CTC. The present study shows that CTC recoveries averaged 35% lower because of epimerization initiated by the use of McBuffer in the extraction procedure.

Animal Feed↗

Study on the fluorescence system of chlortetracycline-Eu-TOPO-sodium dodecyl sulfonate and the determination of chlortetracycline.

The fluorescence system of Eu-chlortetracycline-TOPO-sodium dedecyl sulfonate was studied. It was found that chloretetracycline formed a complex with Eu(III) at pH 8.0-9.0 and then emitted the characteristic fluorescence of Eu(III). TOPO and sodium dodecyl sulfonate greatly enhanced the fluorescence intensity of the system. The experiments indicated that under the optimum determining conditions a linear relationship was obtained between the fluorescence intensity and chlortetracycline concentration in the range of 2.0 x 10(-8)-1.0 x 10(-5) M. The detection limit was 6.0 x 10(-9) M. In addition, the luminescence mechanism of the complex system has been discussed.

Chlortetracycline↗

Analytical procedure for the determination of chlortetracycline and 4-epi-chlortetracycline in pig kidneys.

A liquid chromatography-diode array detection (LC-DAD) procedure has been developed for assaying chlortetracycline (CTC) and its 4-epimer (4-epi-CTC) residues in pig kidneys. The procedure involved extraction with 0.1 M oxalic buffer followed by protein precipitation with trichloroacetic acid. Further solid-phase extraction (SPE) clean-up on a Strata X polymeric cartridge was allowed to obtain an extract suitable for LC analysis. Chromatographic separation was carried out on a C8 analytical column, using isocratic elution with methanol-acetonitrile-0.01 M oxalic acid (15:15:70, v/v/v) at ambient temperature. The flow-rate was 1.2 ml/min and the eluate was analysed at 365 nm. The whole procedure was evaluated according to the requirements of the European Union regulation 2002/657/EC determining specificity, decision limit (CCalpha), detection capacity (CCbeta), trueness, precision and robustness during validation process. The decision limit (CCalpha) was 674.8 microg/kg for CTC and 683.6 microg/kg for 4-epi-CTC. The detection capacity (CCbeta) was 683.6 and 696.3 microg/kg for CTC and 4-epi-CTC, respectively. The recoveries of CTC and 4-epi-CTC from spiked samples at the levels of 300, 600 and 900 microg/kg (0.5 x MRL, 1 x MRL and 1.5 x MRL) were higher than 70%. This method has higher throughput than reported previously extraction method with oxalic acid and acetonitrile used for dechelation and deproteinization.

Animals↗

Iso- and epi-iso-chlortetracycline and the principal metabolites of chlortetracycline in the hen's egg.

Chlortetracycline (CTC) is a broad spectrum antibiotic, licensed for use without any withdrawal period, in chickens laying eggs intended for human consumption. In the European Union, a maximum residue limit (MRL) in eggs of 200 microgram/kg for the sum of the concentrations of CTC and its 4-epimer (4-epi-CTC) has been established. Two major CTC metabolites have been identified in eggs. These compounds, iso-CTC and 4-epi-iso-CTC, have never previously been shown to be significant products of CTC metabolism in poultry or in any other species. The total amount of CTC present in eggs, as all of the chemical forms measured, can exceed the MRL by anything up to a factor of four (170-820 micrograms/kg).

Aluminum↗

Total internal reflected resonance light scattering determination of chlortetracycline in body fluid with the complex cation of chlortetracycline-europium-trioctyl phosphine oxide at the water/tetrachloromethane interface.

A highly selective method of chlortetracycline (CTC) is proposed on the basis of the measurements of total internal reflected resonance light scattering (TMR-RLS) at water/tetrachloromethane (H20/CCl4) interfaces. In the pH range of 7.54-8.14, the interaction of the binary complex of Eu(III)/CTC in the presence of trioctyl phosphine oxide (TOPO) occurs at the H20/CCl4 interface, resulting in greatly enhanced TIR-RLS signals with the maximum peak located at 340 nm. The enhanced TIR-RLS intensity is in proportion to the CTC concentration in the range 0.98 to approximately 20.0 x 10(-7) mol/L. The limit of detection is 9.8 x 10(-9) mol/L. Synthetic samples and body fluid samples including human urine, human serum, and fresh milk were determined with the recovery of 95.4-106.4% and RSD of 2.9-3.9%.

Animals↗

[Determinants of resistance to chlortetracycline and other antibiotics in chlortetracycline-producing strain of Streptomyces aureofaciens].

Data are presented on resistance of Streptomyces aureofaciens strain TB-633 FU--the producer of chlortetracycline (CTC) to autogenous antibiotics and a number of other antibiotics. It is demonstrated that resistance to CTC is specified by ctr genes of constitutive expression as well as by inducible genes. CTC and ethidium bromide may serve as efficient inductors of inducible ctr genes. The induction process is accompanied by increase in antibiotic biosynthesis level. Genes responsible for strain resistance to a number of macrolide antibiotics and thiostrepton are inducible and only function in the presence of appropriate antibiotics in the medium. The action of inducible mtr gene(s) is described in detail. The gene(s) simultaneously ensure increase in resistance to CTC and a number of macrolide antibiotics in the presence of exogenous inductors in media, such as both CTC and macrolide antibiotics. Mutants have been isolated which provide constitutive level of resistance to these antibiotics. A series of ctr and mtr mutants have increased CTC biosynthesis as compared to the initial level. Data on comparative analysis of the results obtained from hybridization of fragments of S. aureofaciens and S. rimosus DNAs to actI and actIII genes, responsible for polyketide synthases' synthesis, demonstrate that genes for CTC and OTC biosynthesis are situated on DNA fragments of similar size. This determines the strategy for cloning ctr and mtr genes as well as genes for CTC biosynthesis from S. aureofaciens.

Chlortetracycline↗

[Molecular cloning of chlortetracycline resistance gene from chlortetracycline producer Streptomyces aureofaciens].

The chlortetracycline (CT) resistance gene ctr was cloned from S. aureofaciens 633, a strain producing the antibiotic. The 6.6-kb DNA Bam HI fragment containing the resistance gene was cloned with the plasmid vector pIJ699. Comparison of the restriction maps of the cloned gene and the oxytetracycline (OT) resistance gene otrA from S. rimosus revealed their similarity which enabled identification of the cloned resistance gene as otrA. Investigation of the resistance determinants in S. aureofaciens 633 made it possible to identify a mtr gene(s). It was demonstrated that introduction of a ctrA gene into S. lividance provided a simultaneous increase in the resistance of the recipient strain to CT and a number of macrolide antibiotics. The CT resistance determinants in S. lividans TK64 showed properties of exogenous induction by CT and the macrolide antibiotics similar to the properties of the mtr gene(s) of S. aureofaciens. Possible adaptation properties of mtr genes are discussed.

Chlortetracycline↗