INTERNATIONAL JOURNAL OF ANTIMICROBIAL AGENTS
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2017
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IF 4.307
Hong-Yan Luo,Ma-Feng Liu,Ming-Shu Wang,Xin-Xin Zhao,Ren-Yong Jia,Shun Chen,Kun-Feng Sun,Qiao Yang,Ying Wu,Xiao-Yue Chen,Francis Biville,Yuan-Feng Zou,Bo Jing,An-Chun Cheng,De-Kang Zhu
The Gram-negative bacterium Riemerella anatipestifer CH-2 is resistant to lincosamides, having a lincomycin (LCM) minimum inhibitory concentration (MIC) of 128 µg/mL. The G148_1775 gene of R. anatipestifer CH-2, designated lnu (H), encodes a 260-amino acid protein with ≤41% identity to other reported lincosamide nucleotidylyltransferases. Escherichia coli Rosetta TM (DE3) containing the pBAD24- lnu (H) plasmid showed four- and two-fold increases in the MICs of LCM and clindamycin (CLI), respectively. A kinetic assay of the purified Lnu(H) enzyme for LCM and CLI showed that the protein could inactive lincosamides. Mass spectrometry analysis demonstrated that the Lnu(H) enzyme catalysed adenylylation of lincosamides. In addition, an lnu (H) gene deletion strain exhibited 512- and 32-fold decreases in LCM and CLI MICs, respectively. The wild-type level of lincosamide resistance could be restored by complementation with a shuttle plasmid carrying the lnu (H) gene. The transformant R. anatipestifer ATCC 11845 [ lnu (H)] acquired by natural transformation also exhibited high-level lincosamide resistance. Moreover, among 175 R. anatipestifer field isolates, 56 (32.0%) were positive for the lnu (H) gene by PCR. In conclusion, Lnu(H) is a novel lincosamide nucleotidylyltransferase that inactivates LCM and CLI by nucleotidylylation, thus conferring high-level lincosamide resistance to R. anatipestifer CH-2.
研究领域:
微生物学
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分子生物学
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抗生素抗性研究
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产品关联
DOI
PubMed
高影响因子
VETERINARY MICROBIOLOGY
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2022
·
IF 3.246
Hongyan Luo,Mengru Li,Zhishuang Yang,Dekang Zhu,Pei Li
A novel chromosomally-located β-lactamase gene, bla RASA-1 , was identified in Riemerella anatipestifer RA-CH-1. The RASA-1, encoded by bla RASA-1 , was a class A extended-spectrum β-lactamase (ESBL), which shared 42.7% and 40.5% identities with the RAA-1 and CGA-1 β-lactamase, respectively. Overexpression of RASA-1 in Escherichia coli confers broad resistance to β-lactams and the purified native RASA-1 revealed ESBL-like hydrolysis activity. Blasting in GenBank showed that bla RASA-1 was exclusively detected in Riemerella anatipestifer . Moreover, sequence analysis revealed that this gene was located within the multi-resistance region of Riemerella anatipestifer genome.
研究领域:
微生物学
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抗药性研究
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DOI
高影响因子
Frontiers in Pharmacology
·
2024
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IF 4.4
Lu Yixing,Qiao Weimei,Xue Yaqian,Hong Xiaoxin,Jin Yuhang,Li Jie,Peng Xianfeng,Zeng Dongping,Zeng Zhenling
Introduction: Riemerella anatipestifer (R. anatipestifer) is an important pathogen in waterfowl, leading to substantial economic losses. In recent years, there has been a notable escalation in the drug resistance rate of R. anatipestifer. Consequently, there is an imperative need to expedite the development of novel antibacterial medications to effectively manage the infection caused by R. anatipestifer.Methods: This study investigated the in vitro and in vivo antibacterial activities of a novel substituted benzene guanidine analog, namely, isopropoxy benzene guanidine (IBG), against R. anatipestifer by using the microdilution method, time-killing curve, and a pericarditis model. The possible mechanisms of these activities were explored.Results and Discussion: The minimal inhibitory concentration (MIC) range of IBG for R. anatipestifer was 0.5–2 μg/mL. Time-killing curves showed a concentration-dependent antibacterial effect. IBG alone or in combination with gentamicin significantly reduced the bacterial load of R. anatipestifer in the pericarditis model. Serial-passage mutagenicity assays showed a low probability for developing IBG resistance. Mechanistic studies suggested that IBG induced membrane damage by binding to phosphatidylglycerol and cardiolipin, leading to an imbalance in membrane potential and the transmembrane proton gradient, as well as the decreased of intracellular adenosine triphosphate. In summary, IBG is a potential antibacterial for controlling R. anatipestifer infections.
研究领域:
微生物学
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兽医学
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药理学
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DOI
PubMed
高影响因子
SCIENCE OF THE TOTAL ENVIRONMENT
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2023
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IF 9.8
Zhishuang Yang,Tianjing Lan,Hongyan Luo,Pei Li,Mingshu Wang,Renyong Jia,Shun Chen,Mafeng Liu,Xinxin Zhao,Qiao Yang,Ying Wu,Shaqiu Zhang,Juan Huang,Xumin Ou,Sai Mao,Qun Gao,Di Sun,Bin Tian,Anchun Cheng,Dekang Zhu
Antimicrobial resistance remains an utmost concern in human and veterinary medicine, impacting humans, animals, and the environment while significantly influencing the principles of One Health. While Riemerella anatipestifer ( R. anatipestifer ) is recognized as a waterfowl pathogen with multidrug-resistant properties, the specifics of its lincosamide resistance mechanism are inadequately understood. In this study, we identified a novel lincosamide resistance gene, lnu (I), in R. anatipestifer RCAD0121, and investigated its potential origin, transfer mechanisms, and dissemination status through genomic epidemiology. This exhibited 74.80 % amino acid identity with a previously reported gene, lnu (H). PCR analysis revealed lnu (I) prevalence in at least 44 R. anatipestifer isolates collected from multiple provinces in China. Furthermore, genomic mining unveiled 56 lnu (I) sequences within publicly available databases, primarily originating from environmental sources. In addition, members of the family Flavobacteriaceae were the dominant (16/56, 28.57 %) bacteria carrying the lnu (I) gene, with Flavobacterium exhibiting a similar GC content as lnu (I). Notably, specific instances of the lnu (I) gene were linked to mobile genetic elements within human and animal pathogenic bacteria. These findings suggest that Flavobacterium species within the environment could serve as potential ancestral sources of the novel lnu (I) gene, which has undergone mobilization events toward pathogenic bacteria.
研究领域:
微生物学
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分子生物学
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医学
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基因组学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF ANTIMICROBIAL CHEMOTHERAPY
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2023
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IF 5.2
Pei Li,Zhishuang Yang,Ting Lei,Yujie Dai,Yang Zhou,Dekang Zhu,Hongyan Luo
Objectives: To elucidate the role of a novel carbapenem-hydrolysing class D β-lactamase (RAD-1) from Riemerella anatipestifer. Methods: We applied WGS and bioinformatic analysis to screen putative β-lactamase genes in R. anatipestifer SCVM0004. A putative class D β-lactamase gene was cloned into pET24a and transferred into Escherichia coli BL21 (DE3) for antibiotic susceptibility determination and protein purification. Meanwhile, the purified native protein was used to determine the enzymatic activities. Results: A class D β-lactamase, RAD-1, was identified in the genome of R. anatipestifer SCVM0004. It was distinct from all characterized class D β-lactamases (≤42% amino acid sequence identity). Searching in GenBank showed that blaRAD-1 was widely disseminated among R. anatipestifer. Genomic environment analysis indicated that the chromosomal structures of blaRAD-1-located regions were relatively conserved. Expression of RAD-1 in E. coli results in elevated MICs for various β-lactam antibiotics, including penicillins, extended-spectrum cephalosporins, a monobactam and carbapenems. Moreover, kinetic analysis of purified RAD-1 revealed: (i) high-level activity against penicillins; (ii) highest affinity for carbapenems; (iii) moderate hydrolysis of extended-spectrum cephalosporins and a monobactam; and (iv) no activity for oxacillin and cefoxitin. Conclusions: This study identified a novel chromosomally located class D carbapenemase RAD-1 (Bush-Jacoby functional group 2def) in R. anatipestifer SCVM0004. Moreover, bioinformatic analysis confirmed that the RAD-1 was widely prevalent and conserved in R. anatipestifer.
研究领域:
微生物学
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基因组学
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抗菌素耐药性
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生物信息学
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DOI
PubMed
高影响因子
BMC MICROBIOLOGY
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2019
·Vol. 19
·期 1
·1-8
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IF 3.287
Zhu Dekang,Zheng Mingyu,Xu Jinge,Wang Mingshu,Jia Renyong,Chen Shun,Liu Mafeng,Zhao Xinxin,Yang Qiao,Wu Ying,Zhang Shaqiu,Huang Juan,Liu Yunya,Zhang Ling,Yu Yanling,Pan Leichang,Chen Xiaoyue,Cheng Anchun
Background Riemerella anatipestifer is one of the most serious infectious disease-causing pathogens in the duck industry. Drug administration is an important method for prevention and treatment of infection in duck production, leading to widespread drug resistance in R. anatipestifer . Methods For a total of 162 isolates of R. anatipestifer, the MICs were determined for a quinolone antimicrobial agent, namely, nalidixic acid, and three fluoroquinolones, namely, ciprofloxacin, enrofloxacin and ofloxacin. The gyr A, par C, and par E gene fragments were amplified by PCR to identify the mutation sites in these strains. Site-directed mutants with mutations that were detected at a high frequency in vivo were constructed (hereafter referred to as site-directed in vivo mutants), and the MICs of these four drugs for these strains were determined. Results In total, 100, 97.8, 99.3 and 97.8% of the 137 R. anatipestifer strains isolated between 2013 and 2018 showed resistance to nalidixic acid, ciprofloxacin, enrofloxacin, and ofloxacin, respectively. The high-frequency mutation sites were detected in a total of 162 R. anatipestifer strains , such as Ser83Ile and Ser83Arg, which are two types of substitution mutations of amino acid 83 in GyrA; Val799Ala and Ile811Val in ParC; and Val357Ile, His358Tyr, and Arg541Lys in ParE. MIC analysis results for the site-directed in vivo mutants showed that the strains with only the Ser83Ile mutation in GyrA exhibited an 8–16-fold increase in MIC values, and all mutants showed resistance to ampicillin and ceftiofur. Conclusions The resistance of R. anatipestifer to quinolone agents is a serious problem. Amino acid 83 in GyrA is the major target mutation site for the fluoroquinolone resistance mechanism of R. anatipestifer .
研究领域:
兽医学
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微生物学
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分子生物学
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DOI
高影响因子
危险品化学品经营许可证（带存储）