ESBLs-producing Escherichia coli from sheep-origin: Genome-wide virulence genes identification and in vivo virulence assessment in mice and Galleria mellonella.

The worldwide spread of pathogenic Escherichia coli, together with the multidrug resistant linked with extended-spectrum ß-lactamases (blaCTX-M , blaTEM and blaOXA ), not only affect the health of animals and humans but also bring huge economic losses to animal husbandry. Despite the high levels of virulence present in many extended-spectrum beta-lactamases (ESBLs)-producing E. coli isolates, however, few studies have comprehensively assessed the pathogenicity of ESBLs-producing E. coli isolates. Thus, the aim of the present study was to investigate the presence of virulence genes in third-generation cephalosporin-resistant E. coli and to assess their pathogenicity and zoonotic potential. Previously, we identified 67 ESBLs-producing E. coli strains from sheep anal swabs in northwest China. In this study, we genotypically and phenotypically characterized isolates of E. coli that produce ESBLs. According to the VirulenceFinder and virulence factors database, all ESBLs-producing E. coli strains harboured a wide range of virulence genes. The ColV plasmid-related genes (hlyF, ompT, iss, iutA and cvaC) were present in 52 (77.6%) ESBLs-producing E. coli isolates. Surprisingly, quite a number of extraintestinal pathogenic E. coli virulence-related genes were detected in 62 (92.5%) of 67 isolates. A total of 33 serotypes and 37 sequence types (STs) were found in 67 ESBLs-producing isolates. ST10 is the most prevalent ST, which is represented by five strains. The cluster analysis showed that CC10 and CC23 were the common clonal complexes (CCs). Predominant serotypes were O8 (10%) and O9 (9%) followed by 6% each of O89, O101 and O185. Most sheep-origin ESBLs-producing E. coli held the highly pathogenic to human and displayed moderate-to-vigorous-intensity motor capacity. The ESBLs-producing E. coli isolates with numerous virulence-related genes were able to cause multiple infectious diseases in animal models (mice, neonatal rats and Galleria mellonella). To our knowledge, this study represents an important first step for a comprehensive characterization of pathogenicity and zoonotic potential of sheep-origin ESBLs-producing E. coli isolates. These findings may be of significant value for the identification of pathogenicity and zoonotic potential risks associated with sheep-origin ESBLs-producing E. coli.

Impact of l-Arginine and l-Histidine on the structural, optical and antibacterial properties of Mg doped ZnO nanoparticles tested against extended-spectrum beta-lactamases (ESBLs) producing Escherichia coli.

Magnesium doped Zinc oxide nanoparticles (Mg:ZnO NPs) were synthesized by co-precipitation method. The synthesized Mg:ZnO NPs exhibited hexagonal wurtzite structure, which was confirmed by X-ray diffraction results. After structural confirmation of Mg doped ZnO NPs, base amino acids like l-Arginine and l-Histidine were separately incorporated with the Mg: ZnO NPs. l-Arginine added Mg:ZnO (Mg:ZnO:LA) and l-Histidine added Mg:ZnO (Mg:ZnO: LH) NPs retained the same wurtzite hexagonal structure and average crystallite sizes of Mg: ZnO:LA and Mg: ZnO:LH NPs were found to be 25 nm and 20 nm respectively. The sizes of Mg:ZnO:LH and Mg: ZnO: LA NPs decreased as compared to that of the Mg doped ZnO NPs. From the FT-IR spectra, the ZnO stretching frequencies were observed at 516, 517 and 518 cm-1 for Mg:ZnO, Mg:ZnO: LA and Mg: ZnO:LH NPs respectively. From the FESEM images, the morphologies of ZnO:Mg and ZnO:Mg:LA NPs were spherical and the Mg: ZnO: LH NPs formed nano-flakes structure. From the EDAX study, the amount of elements incorporated in the samples was determined. The photoluminescence measurements revealed the existence of zinc vacancies, oxygen vacancies and surface defects of the samples. Antibacterial activity of the amino acid added Mg doped ZnO NPs was studied against extended-spectrum beta-lactamases (ESBLs) producing Escherichia coli (E. coli).The Minimal Inhibitory Concentration (MIC) of the LH added ZnO:Mg NPs was found to be 1000 µg/ml for which the growth of E. coli completely inhibited. l-Histidine added Mg doped ZnO NPs showed the highest antibacterial activity as compared to that of the Mg:ZnO NPs and ZnO:Mg:LA NPs.

Bacterial surveillance and drug resistance analysis of ESBLs Enterobacteriaceae produced by female vaginal secretions / 国际检验医学杂志

Objective In order to understand the distribution and drug resistance of the extended-spectrumβ-lactamase-producing (ESBLs) Enterobacteriaceae in female vaginal secretions and to provide the basis for clinical treatment.Methods 939 strains of Enterobacteriaceae isolated from female vaginal secretions were collected from Obstetrics and Gynecology Hospital of Fudan University from January 2014 to December 2017.The strain identification and drug sensitivity test of VITECK 2 Compact-totally automatic bacterial identification analyzer were used to analyze the detection rate and drug resistance of ESBLs Enterobacteriaceae.Results 257 strains of ESBLs-producing strains were detected in 939 strains of Enterobacteriaceae with a detection rate of 27％, including 220 Escherichia coli, 34 Klebsiella pneumoniae, 2 Stink-nose Klebsiella and 1 strain of Acidogenic Klebsiella.ESBLs Escherichia coli and Klebsiella pneumoniae were all resistant to Ampicillin and Cefazolin, and to Ceftazidime, Nitrofurantoin, SMZ, Amikacin, Ciprofloxacin, Gentamicin, tobramycin, Ampicillin/Sulbactam, Ceftriaxone, Ertapenem, Piperacillin/Tazobactam, Aztreonam, Cefepime, Levofloxacin the drug resistance rates of were 36％and 44％, 2％ and 41％, 67％ and 68％, 2％ and 15％, 58％ and 38％, 51％ and65％, 14％and 26％, 65％and 76％, 99％and 97％, 0％and 29％, 0％and 3％, 50％and 65％, 26％and 18％, 57％ and 32％, but they are all sensitive to Imipenem and Cefotetan.Conclusion The inicdence of ESBLs Enterobacteriaceae in female vaginal secretions is high, and Imipenem, Cefotetan, Piperacillin/Tazobactam have high antibacterial activity, which can be used as the experience of initial treatment.

Biologically synthesized zinc oxide nanoparticles as nanoantibiotics against ESBLs producing gram negative bacteria.

The accelerative outgrowth of extended spectrum ß-lactamases (ESBLs) producing Escherichia coli (E. coli) and Proteus mirabilis (P. mirabilis) was mainly due to incessant relentless influence of antibiotics thereby increasing incidence and death rate which was obvious from the survey of ESBLs producing bacteria related health problem. In the present paper, we synthesized and characterized zinc oxide nanoparticles (ZnO NPs) employing using Camellia japonica leaf extract, bactericidal action of these NPs against extended spectrum ß lactamases (ESBLs) positive E. coli and P. mirabilis clinical strains owing the minimal inhibitory concentration (MIC) percentage 83, 81% at 100 µg/mL concentration and minimum bactericidal concentration (MBC) final inhibiting concentration at 150 µg/mL. Moreover, confocal laser scanning microscopy (CLSM) and scanning electron microscope (SEM) results evident for loss of viability, cell shrinkage, disarrangement of cell membrane, and cell wall lysis activity of ZnO NPs against ESBLs positive E. coli BDUMS3 (KY617770) and P. mirabilis BDUMS1 (KY617768) strains. From the results, it was observed that the biologically synthesized ZnO NPs has stronger antibacterial effect against ESBLs producing bacterial strains. Nevertheless, current date there is no reports of antibacterial activity of metal oxide (ZnO) NPs against ESBL producing gram negative bacteria. Consequently, this finding is the first report in this respect and it shows band gap energy and ROS accumulation to damage the cell wall and inhibit the growth of ESBL producing gram negative strains.

Pharmaceutical Care for a Case of ESBLs Producing Klebsiella pneumoniae Severe Pneumonia / 中国药房

OBJECTIVE: To probe the role of clinical pharmacists in drug use of patients with severe pneumonia. METHODS: Clinical pharmacists participated in the treatment for a patient with ESBLs producing Klebsiella pneumoniae severe pneumonia. The patient was given cefoperazone tazobactam combined with moxifloxacin and ganciclovir initially. Clinical pharmacists suggested stopping cefoperazone tazobactam and moxifloxacin and additionally using meropenem according to the elevation of hemogram infection indexes; suggested stopping ganciclovir and continuously using meropenem according to the results of sputum culture; suggested providing cefoperazone sulbactam de-escalation sequential therapy for ESBLs producing Klebsiella pneumoniae and stopping cefoperazone sulbactam and azithromycin according to clinical symptoms of the patient. RESULTS: The physicians adopted the suggestions of clinical pharmacists. After treatment, body temperature and lab indexes of the patient recovered to normal; the result of sputum culture turned to negative. Chest CT showed that the infection focus was obviously absorbed compared to before. After discharged from hospital and followed up, the patient was found to have a good prognosis. CONCLUSIONS: Through actively participating in drug therapy, based on lab indexes and results of sputum culture, clinical pharmacists provide pharmaceutical care and adjust medication plan to improve treatment rate of patients with severe pneumonia and the safety and effectiveness of drug treatment, be of great significance to promote the rational use of antibiotics.

Study on integron drug resistance gene of ESBLs-prodcing Klebsiella pneumonia infection in ICU elderly patients / 国际检验医学杂志

Objective To understand the gene distribution and drug resistance rate of integron gene of extended spectrumβ‐lac‐tamases(ESBLs) producing Klebsiella pneumonia infection in ICU elderly patients in order to provide the basis for rational use of antimicrobial agents .Methods The BioMerieux VITEK‐2 Automated Microbes Identification System was adopted to conduct the bacteria identification and drug susceptibility test on various clinical specimens of ICU elderly patients in our hospital from January 2013 to December 2015 .The integron gene in 167 strains of ESBLs‐producing Klebsiella pneumoniae was analyzed by PCR ,and the gene was identified by sequencing .Results Among 386 strains of Klebsiella pneumoniae ,the detection rate of ESBLs‐producing strains was 43 .26% ;the positive rate of integron was 50 .89% ,the detected integron was class Ⅰ integron;aadA2 ,aadA1 ,aada16 , dfra27 and arr‐3 genes were amplified from integron variable region;the drug resistance rate of ESBLs‐producing integron gene pos‐itive strains was significantly higher than that of integron gene negative strains .Conclusion the ICU elderly patients with ESBLs‐producing Klebsiella pneumoniae infection is closely related to the integron gene and integron plays an important role in bacterial drug resistance .

Comparison of extended spectrum ß-lactamases-producing Escherichia coli with non-ESBLs-producing E.coli: drug-resistance and virulence.

BACKGROUND: The virulent factors of Escherichia coli (E.coli) play an important role in the process of pathopoiesis. The study aimed to compare drug-resistant genes and virulence genes between extended spectrum ß-lactamases (ESBLs)-producing E.coli and non-ESBLs-producing E.coli to provide a reference for physicians in management of hospital infection. METHODS: From October 2010 to August 2011, 96 drug-resistant strains of E.coli isolated were collected from the specimens in Qingdao Municipal Hospital, Qingdao, China. These bacteria strains were divided into a ESBLs-producing group and a non-ESBLs-producing group. Drug sensitivity tests were performed using the Kirby-Bauer (K-B) method. Disinfectant gene, qacEΔ1-sull and 8 virulence genes (CNF2, hlyA, eaeA, VT1, est, bfpA, elt, and CNF1) were tested by polymerase chain reaction (PCR). RESULTS: Among the 96 E.coli isolates, the ESBLs-producing E.coli comprised 46 (47.9%) strains and the non-ESBLs-producing E.coli consisted of 50 (52.1%) strains. The detection rates of multiple drug-resistant strain, qacEΔ1-sull, CNF2, hlyA, eaeA,VT1, est, bfpA, elt, and CNF1 in 46 ESBLs-producing E.coli isolates were 89.1%, 76.1%, 6.5%, 69.6%, 69.6%, 89.1%, 10.9%, 26.1%, 8.7%, and 19.6%, respectively. In the non-ESBLs-producing E.coli strains, the positive rates of multiple drug-resistant strain, qacEΔ1-sull, CNF2, hlyA, eaeA, VT1, est, bfpA, elt, and CNF1 were 62.0%, 80.0%, 16.0%, 28.0%, 64.0%, 38.0%, 6.0%, 34.0%, 10.0%, and 24.0%, respectively. The difference in the detection rates of multiple drug-resistant strain, hlyA and VT1 between the ESBLs-producing E.coli strains and the non-ESBLs-producing E.coli strains was statistically significant (P<0.05). CONCLUSION: The positive rate of multiple drug-resistant strains is higher in the ESBLs-producing strains than in the non-ESBLs-producing strains. The expression of some virulence genes hlyA and VT1 varies between the ESBLs-producing strains and the non-ESBLs-producing strains. Increased awareness of clinicians and enhanced testing by laboratories are required to reduce treatment failures and prevent the spread of multiple drug-resistant strains.

Comparison of extended spectrum β-lactamases-producing Escherichia coli with non-ESBLs-producing E.coli: drug-resistance and virulence / 世界急诊医学杂志（英文）

@#BACKGROUND: The virulent factors ofEscherichia coli (E.coli) play an important role in the process of pathopoiesis. The study aimed to compare drug-resistant genes and virulence genes between extended spectrum β-lactamases (ESBLs)-producingE.coli and non-ESBLs-producing E.coli to provide a reference for physicians in management of hospital infection. METHODS: From October 2010 to August 2011, 96 drug-resistant strains ofE.coli isolated were colected from the specimens in Qingdao Municipal Hospital, Qingdao, China. These bacteria strains were divided into a ESBLs-producing group and a non-ESBLs-producing group. Drug sensitivity tests were performed using the Kirby-Bauer (K-B) method. Disinfectant gene, qacEΔ1-sull and 8 virulence genes (CNF2, hlyA, eaeA, VT1, est, bfpA, elt, and CNF1) were tested by polymerase chain reaction (PCR). RESULTS: Among the 96E.coli isolates, the ESBLs-producingE.coli comprised 46 (47.9％) strains and the non-ESBLs-producingE.coli consisted of 50 (52.1％) strains. The detection rates of multiple drug-resistant strain, qacEΔ1-sull, CNF2, hlyA, eaeA,VT1, est, bfpA, elt, and CNF1 in 46 ESBLs-producingE.coli isolates were 89.1％, 76.1％, 6.5％, 69.6％, 69.6％, 89.1％, 10.9％, 26.1％, 8.7％, and 19.6％, respectively. In the non-ESBLs-producingE.coli strains, the positive rates of multiple drug-resistant strain, qacEΔ1-sull, CNF2, hlyA, eaeA, VT1, est, bfpA, elt, and CNF1 were 62.0％, 80.0％, 16.0％, 28.0％, 64.0％, 38.0％, 6.0％, 34.0％, 10.0％, and 24.0％, respectively. The difference in the detection rates of multiple drug-resistant strain, hlyA and VT1 between the ESBLs-producingE.coli strains and the non-ESBLs-producingE.coli strains was statistically significant (P<0.05). CONCLUSION: The positive rate of multiple drug-resistant strains is higher in the ESBLs-producing strains than in the non-ESBLs-producing strains. The expression of some virulence genes hlyA and VT1 varies between the ESBLs-producing strains and the non-ESBLs-producing strains. Increased awareness of clinicians and enhanced testing by laboratories are required to reduce treatment failures and prevent the spread of multiple drug-resistant strains.

Detection of ESBLs Produced by E. coli in Urinary Tract and Monitoring of Drug Resistance of ESBLs-producing E.coli / 中国药房

OBJECTIVE:To investigate the condition of ESBLs produced by E.coli isolated from urinary tract and the drug resistance of ESBLs-producing E.coli.METHODS:The identification of bacteria was performed using ATB-Expression analysator(France);the susceptibility test was performed using K-B method,and ESBLs were detected using disc diffusion confirmatory test.RESULTS:The detection rate of ESBLs-producing E.coli was 31.8%.All(100%)of the 107 strains of ESBLs-producing E.coli were sensitive to imipenem,however,in which different degree of resistance to other antibiotics was noted.The resistance rate was significantly higher in ESBLs-producing strains than in non-ESBLs-producing strains.CON-CLUSION:In view of the high antibiotic resistance of ESBLs-producing E.coli,great importance should be attached to the detection of the ESBLs.Antibiotics should be used rationally based on the results of susceptibility test.