Bacterial Dehydrogenases Facilitate Oxidative Inactivation and Bioremediation of Chloramphenicol

Antimicrobial resistance represents a major threat to human health and knowledge of the underlying mechanisms is therefore vital. Here, we report the discovery and characterization of oxidoreductases that inactivate the broad-spectrum antibiotic chloramphenicol via dual oxidation of the C3-hydroxyl group. Accordingly, chloramphenicol oxidation either depends on standalone glucose-methanol-choline (GMC)-type flavoenzymes, or on additional aldehyde dehydrogenases that boost overall turnover. These enzymes also enable the inactivation of the chloramphenicol analogues thiamphenicol and azidamfenicol, but not of the C3-fluorinated florfenicol. Notably, distinct isofunctional enzymes can be found in Gram-positive (e. g., Streptomyces sp.) and Gram-negative (e. g., Sphingobium sp.) bacteria, which presumably evolved their selectivity for chloramphenicol independently based on phylogenetic analyses. Mechanistic and structural studies provide further insights into the catalytic mechanisms of these biotechnologically interesting enzymes, which, in sum, are both a curse and a blessing by contributing to the spread of antibiotic resistance as well as to the bioremediation of chloramphenicol.     

乳和乳制品中残留抗生素的检测方法

论述了牛奶中残留抗生素的原因及危害性，列举了目前世界上较为流行的牛奶抗生素检测方法，并介绍了其检测原理。     

动物食品中四环素残留的研究进展

养殖业上,四环素类药物在促进畜禽生长、提高饲料报酬、疾病防治等方面发挥着重要作用。但是养殖户对四环素类药物的应用状况非常混乱,滥用四环素类药物的情况时有发生。由于四环素的滥用会给人和动物的健康带来危害,同时对动物性食品的食用安全带来不稳定因素。因此对四环素类药物残留的研究就显得尤为重要。本文主要阐述了四环素残留对人体健康的危害,四环素残留研究状况,四环素残留的一些相关的国家标准。最后提出四环素残留有待研究的问题和意义。     

毛细管电泳法分析食品中残留抗生素的研究进展

食品中残留的抗生素是影响食品安全质量的因素之一,严重影响着动物类食品的质量。综述毛细管电泳技术检测方法的特点,以及应用该法检测食品中β-内酰胺类、大环内酯类、氨基糖苷类、四环类等几种抗生素的研究进展。     

液相色谱-质谱联用检测乳中β-内酰胺类抗生素研究进展

β-内酰胺类抗生素是畜牧和牛乳生产中最广泛应用的、治疗动物细菌感染的抗生素。由于它们常用作治疗细菌感染的兽药,因此对于动物性食品和乳中β-内酰胺类抗生素的残留量分析是很重要的。论述了在检测牛乳中β-内酰胺类抗生素中的最新应用状况,表明液相色谱-质谱联用法(LC-MS)是一种高灵敏度和选择性的检测方法,具有广阔的发展前景。     

Alanine Scan of the Peptide Antibiotic Feglymycin: Assessment of Amino Acid Side Chains Contributing to Antimicrobial Activity

The antibiotic feglymycin is a linear 13‐mer peptide synthesized by the bacterium Streptomyces sp. DSM 11171. It mainly consists of the nonproteinogenic amino acids 4‐hydroxyphenylglycine and 3,5‐dihydroxyphenylglycine. An alanine scan of feglymycin was performed by solution‐phase peptide synthesis in order to assess the significance of individual amino acid side chains for biological activity. Hence, 13 peptides were synthesized from di‐ and tripeptide building blocks, and subsequently tested for antibacterial activity against Staphylococcus aureus strains. Furthermore we tested the inhibition of peptidoglycan biosynthesis enzymes MurA and MurC, which are inhibited by feglymycin. Whereas the antibacterial activity is significantly based on the three amino acids D ‐Hpg1, L ‐Hpg5, and L ‐Phe12, the inhibitory activity against MurA and MurC depends mainly on L ‐Asp13. The difference in the position dependence for antibacterial activity and enzyme inhibition suggests multiple molecular targets in the modes of action of feglymycin.     

Pilot survey of hen eggs consumed in the metropolitan area of Rio de Janeiro,Brazil, for polyether ionophores,macrolides and lincosamides residues

A liquid chromatography–tandem mass spectrometry (LC–MS/MS) method, which has recently been developed and validated, was used for the identification and quantification of polyether ionophore, macrolide and lincosamide residues in commercial eggs sold in the metropolitan area of Rio de Janeiro, Brazil. The method was applied to 100 samples and the results showed a high incidence of polyether ionophore residues (25%). Salinomycin was detected in 21% of samples, but only two non-compliant results (5.3 and 53 µg?kg^?1) were found if maximum limits (tolerances) established by European Union were adopted in Brazil and if a method decision limit (CCα) of 3.4?µg?kg^?1 was considered. In 8% of analyzed samples, more than one studied coccidiostat was found. The lincosamide, lincomycin, and the macrolide, tylosin, were detected at trace levels in 4 and 1% of the samples, respectively. Lasalocid, clarithromycin and erythromycin were not found.     

Analysis of sulphonamide residues in edible animal products: A review

The methods of analysis for sulphonamide residues in edible animal products are reviewed. Sulphonamides are widely used for therapeutic and prophylactic purposes in both humans and animals, sometimes as growth promoters as additives in animal feed. As a result of their widespread use, there is concern about whether the levels used of these drugs can generate serious problems in human health, e.g., allergic or toxic reactions. Several methods for the determination of sulphonamides have been reported in the literature and this review considers high-performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC/MS), gas chromatography (GC), thin-layer chromatography (TLC), high-performance capillary electrophoresis (HPCE), enzyme-linked immunosorbant assay (ELISA), biosensor immunoassay (BIA) and microbiological methods. Specific aspects of analysing sulphonamides, such as sample handling, chromatographic conditions and detection methods are discussed. Methods for drug residue monitoring should be accurate, simple, economical in both time and cost, and capable of detecting residues below the maximum residue limits (MRL). The current sulphonamide detection technologies are based on chromatographic methods or bacteriological growth inhibition. The instrumental methods such as HPLC and GC are both sensitive and specific, but are laborious and expensive. Because of the labour-intensive processes, only a few cases of GC methods applied to residue analysis have been published. These methods are suitable for confirmation but not for screening of large numbers of samples. Microbiological methods do not require highly specialized and expensive equipment. They also use highly homogeneous cell populations for testing and thus result in better assay precision. Although HPCE has powerful separation ability, the precision is poor and the instrument still needs to be improved. To date, this technique has not been widely applied to routine analysis. Currently, TLC has been almost replaced by other instrumental analysis. A rapid, sensitive and specific assay is required to detect positive samples in routine analysis, which can then be confirmed for the presence of sulphonamides by HPLC. Immunochemical methods such as ELISA can be simple, rapid and cost-effective, with enough sensitivity and specificity to detect small molecules. This review can be considered as a basis for further research aimed at identifying the most efficient approaches.     

(3-烯丙氧基苯基氨基羰基)-N-对硝基苄氧基羰基-反-4-羟基-L-吡咯烷的合成研究

以4R-羟基-L-脯氨酸为原料,与氯甲酸对硝基苄酯反应进行氨基保护、再与间氨基苯甲酸烯丙酯缩合得到目标化合物(3-烯丙氧基苯基氨基羰基)-N-对硝基苄氧基羰基-反-4-羟基-L-吡咯烷。对反应物配比、催化剂、反应时间和温度等因素进行了研究,确定了缩合优化条件为:二氯甲烷作溶剂,25℃下反应4 h。优化条件下,缩合收率达86.1%。目标产物结构经IR、1H NMR和MS确证。     

Phytol Derivatives as Drug Resistance Reversal Agents

Phytol was chemically transformed into fifteen semi‐synthetic derivatives, which were evaluated for their antibacterial and drug resistance reversal potential in combination with nalidixic acid against E. coli strains CA8000 and DH5α. The pivaloyl ( 4 ), 3,4,5‐trimethoxybenzoyl ( 9 ), 2,3‐dichlorobenzoyl ( 10 ), cinnamoyl ( 11 ), and aldehyde ( 14 ) derivatives of phytol ((2E,7R,11R)‐3,7,11,15‐tetramethyl‐2‐hexadecen‐1‐ol) were evaluated by using another antibiotic, tetracycline, against the MDREC‐KG4 clinical isolate of E. coli. Derivative 4 decreased the maximal inhibitory concentration (MIC) of the antibiotics by 16‐fold, while derivatives 9 , 10 , 11 , and 14 reduced MIC values of the antibiotics up to eightfold against the E. coli strains. Derivatives 4 , 9 , 10 , 11 , and 14 inhibited the ATP‐dependent efflux pump; this was also supported by their in silico binding affinity and down‐regulation of the efflux pump gene yojI, which encodes the multidrug ATP‐binding cassette transporter protein. This study supports the possible use of phytol derivatives in the development of cost‐effective antibacterial combinations.