Extended-spectrum β-lactamase controversies

G.A.Jacoby 教授毕业于美国哈佛医学院,为国际知名的传染病学、微生物学和免疫学专家,曾长期担任美国麻省总医院传染病科医师,现任麻省 Lahey 医学中心传染病实验室主任,中国感染与化疗杂志特邀编委。长期以来 Jacoby 教授对细菌耐药机制有深入研究,尤其对细菌(肺炎克雷伯菌、大肠埃希菌等)产生质粒介导的超广谱β内酰胺酶(ESBL)以及喹诺酮类的耐药机制研究。曾发表多篇有关上述专题的权威性论著。此次本刊特邀 Jacoby 教授撰写有关 ESBL 的新进展,文中阐述目前 ESBLs 除通常所指质粒介导β内酰胺酶中扩大了酶水解的底物谱导致细菌对头孢噻肟、头孢他啶、氨曲南等抗生素耐药外,还包括许多具有不同特点的β内酰胺酶,其中某些酶产自共生菌。除 ESBL 外,AmpC 酶和各种碳青霉烯β内酰胺酶也可导致细菌对上述抗生素耐药,因此临床微生物实验室准确检出和鉴别各种β内酰胺酶,对于临床选用适宜的抗茵药十分重要。美国 CLSI(过去称为 NCCLS)推荐采用两步法(筛选及确证)检测细菌产 ESBL;但有的学者认为测定抗茵药物对细菌的 MIC,如用药后 T>MIC 在50%以上即可达到满意疗效,无需检测细菌是否产 ESBL。目前认为碳青霉烯类抗生素治疗产 ESBL 菌感染的疗效最为满意,但由此可能引起细菌对该类药物耐药,值得关注。采用大剂量头孢吡肟或哌拉西林-三唑巴坦治疗产ESBL 菌感染是否有效尚有争论,临床上对β内酰胺类最为耐药的菌株往往对几乎所有现用抗菌药耐药。黏菌素(或多黏菌素 B)曾成功的用于治疗此种多重耐药菌感染。此外替莫西林(temocillin,一种对β内酰胺酶稳定的青霉素类)与替加环素(tigecvcline,为米诺环素衍生物)体外对产 ESBL 菌有抗菌作用,但尚无临床研究资料。

Post-appendectomy pelvic abscess with extended-spectrum beta-lactamase producing Escherichia coli : A case report and review of literature

BACKGROUND Appendicitis, the inflammation of the appendix, is the most common abdominal surgical emergency requiring expedient surgical intervention. Extendedspectrum beta-lactamases(ESBLs) are bacterial enzymes that catalyse the degradation of the betalactam ring of penicillins and cephalosporins(but without carbapenemase activity), leading to resistance of these bacteria to beta-lactam antibiotics. Recent increases in incidence of ESBL-producing bacteria have caused alarm worldwide. Proportion estimates of ESBLEnterobacteriaceae hover around 46% in China, 42% in East Africa, 12% in Germany, and 8% in the United States.CASE SUMMARY The impact of ESBL-producing bacteria on appendiceal abscesses and consequent pelvic abscesses are yet to be examined in depth. A literature review using the search words "appendiceal abscesses" and "ESBL Escherichia coli(E. coli)" revealed very few cases involving ESBL E. coli in any capacity in the context of appendiceal abscesses. This report describes the clinical aspects of a patient with appendicitis whodeveloped a postoperative pelvic abscess infected with ESBL-producing E. coli. In this report, we discuss the risk factors for contracting ESBL E. coli infection in appendicitis and post-appendectomy pelvis abscesses. We also discuss our management approach for postappendectomy ESBL E. coli pelvic abscesses, including drainage, pathogen identification, and pathogen characterisation. When ESBL E. coli is confirmed, carbapenem antibiotics should be promptly administered, as was done efficaciously with this patient. Our report is the first one in a developed country involving ESBL E. coli related surgical complications in association with a routine laparoscopic appendectomy.CONCLUSION Our report is the first involving ESBL E. coli and appendiceal abscesses, and that too consequent to laparoscopic appendectomy.

Detection of Beta-Lactamase and Extended-Spectrum Beta-Lactamase of Pathogens Isolated from Pig and Chicken and Their Antibiotic Susceptibility Test

The antibacterial activity of beta-lactam antibiotics or their combinations with inhibitor sulbactum against non-lactamase- producing strains, lactamase-producing and ESBLs-producing isolates was evaluated with twofold dilution method after pathogens isolated from pigs and chickens were detected, respectively, for beta-lactamase and extended-spectrum beta- lactamases （ESBLs）, The results revealed that most of 43 clinically isolated strains could produce beta-lactamase and 3 strains of shigella isolated from chicken samples produced ESBLs. All of 30 lactamase-producing strains isolated and only one of 16 non-lactamase-producing strains were resistant to amoxicillin and ampicillin. MICs of ampicillin against lactamaseproducing isolates decreased 10-40 and 10-20 times respectively, when it was conbined with sulbactam at ration of 1：2 and 1：4. All clinical isolates were susceptible to third-generation cephalosporins. The MICs of third-generation cephalosporins against lactamase-producing isolates did not change when they were conbined with sulbactam. MICs of ceftiofur and ceftriaxone against ESBLs-producing isolates decreased 2-4 times when they were conbined with sulbactam.

Appendical Perforation by Infection with Extended-Spectrum Beta-Lactamase (ESBL)-Producing <i>Escherichia coli</i>: Case Report

We report the very rare case of a huge appendical abscess with extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli (E. coli) as the pathogen. There have been several reports of appendical infections such as appendicitis and appendical abscess caused by ESBL-producing bacteria in adults. The treatment of ESBL-producing E. coli infection is specific, and ESBL-producing bacteria have recently been reported as pathogens associated appendicitis in children. To the best of our knowledge, this is the second report of perforated appendicitis with abscess due to ESBL-producing E. coli. We discuss the diagnostic modalities and treatments for appendical abscess with ESBL-producing E. coli. and propose that the patients with perforated appendicitis and abscess formation due to ESBL-producing E. coli should be administered the antibiotic MEPM within 2 weeks to treat the abscess more effec-tively without producing other multidrug-resistant bacteria.

Distribution of extended-spectrum β-lactamase genes in antibiotic-resistant strains of Pseudomonas aeruginosa obtained from burn patients in Ahvaz, Iran

Objective: To evaluate the drug susceptibility profiles and the frequency of beta-lactamase encoding genes in Pseudomonas aeruginosa (P. aeruginosa) obtained from burn patients. Methods: Totally 93 non-duplicate clinical isolates of P. aeruginosa were recovered from burn patients of Taleghani Burn Hospital of Ahvaz. Antibiotic susceptibility testing was conducted by disk diffusion method according to the CLSI 2017 recommendations. PCR assay was performed by to find beta-lactamase encoding genes. Results: In this study, most clinical specimen was obtained via wound swabs [65 (69.9%)], followed by blood [14 (15.1%)] and biopsy (7 (7.5%))Forty-two (45.16%) patients were male and 51(54.84%) were female. High resistance was observed for most of antibiotics especially for gentamicin and ciprofloxacin (Up to 85%), whereas the highest susceptibility was reported for colistin (100.0%), followed by ceftazidime (66.7%). According to PCR results, 16.1% (15), 9.7% (9) and 14.0% (13) of isolates carried blaDHA, blaVEB and blaGES genes, respectively. It also revealed that the blaVEB gene was found to coexist within 2 isolates (2.2%). Conclusions: Antibacterial resistance is high among P. aeruginosa isolates. Colistin is highly active against multi-drug resistant P. aeruginosa isolates. Antimicrobial susceptibility testing can confine indiscriminate uses of antibiotics and resistance increase, and can improve management of treatment.

Relationship of multidrug-resistant gene and extended-spectrum carbapenem-resistance in Staphylococcus aureus

The aim of this study was to determine the relationship between phenotypic antimicrobial susceptibility patterns and extended-spectrum,carbapenem-resistance genes.A total of 109 clinical Staphilococcus aureus strains were subjected to 19 antimicrobial susceptibility tests.Resistance to methicillin(mecA),penicillin(blaTEM),and tetracycline(tetM)was detected.We compared the presence of the blaTEM genes with extended-spectrum,carbapenem-related genes and identified the types of SCCmec genes.Of 109 clinical S.aureus strains,62(56.88%)had methicillin resistance and 60 strains carried mecA.The prevalence of blaTEM and tetM genes was 81.65%and 37.61%,respectively.The most predominant SCCmec type was SCCmec type Ⅱ 28/60(46.67%),in 60 mecA-positive methicillin-resistant S.aureus(MRSA)isolates.The SCCmec prevalence rates were type ⅣA 30.00%(18/60),type Ⅳb 8.33%(5/60),type Ⅳd 6.67%(4/60),and non-typable 8.33%(5/60).Sixty of the 109(55.05%)MRSA isolates were positive for extended-spectrum carbapenems(31/60)(51.67%),cephalosporins 40/60(66.67%)and carbapenems 31/60(51.67%).The predominant SCCmec type II demonstrated more carbapenem-resistance than the ⅣA,Ⅳb and Ⅳd types.

Antibiogram of Extended-Spectrum <i>β</i>-Lactamase (ESBL) Producing <i>Escherichia coli</i>

Background: Extended-spectrum β-lactamases (ESBLs) are enzymes capable of hydrolyzing extended-spectrum cephalosporins, penicillins and monobactams but inactive against cephamycins and carbapenems. The ESBL-producing organisms are a breed of multidrug-resistant pathogens. Objectives: This study was aimed to determine the susceptibility pattern of ESBL-producing Escherichia coli to ciprofloxacin, amikacin and imipenem. Methods: A total of 75 ESBL-producing E. coli, were obtained from the tertiary care hospitals of Bangladesh and were studied for susceptibility pattern from October, 2010 to December, 2011. These isolates were identified by double disc synergy test (DDST) and were confirmed phenotypically as ESBL-producer by phenotypic confirmatory disc diffusion test (PCDDT). Minimum inhibitory concentrations (MICs) of ciprofloxacin, amikacin and imipenem among ESBL-producing E. coli were determined using agar dilution method. Results: Out of 75 DDST positive ESBL-producing E. coli, 71 (94.67%) were also positive by PCDDT. All ESBL-producing E. coli, were susceptible to imipenem. About 92.95% ESBL-producing E. coli were susceptible to amikacin but only 14.08% were susceptible to ciprofloxacin. Conclusion: In this study, ESBL-producing E. coli, showed high resistance to ciprofloxacin. Imipenem and amikacin were most effective against ESBL positive strains.

Microbiologic and Clinical Comparison of Patients Harboring <i>Escherichia coli</i>Blood Isolates with and without Extended-Spectrum <i>β</i>-Lactamases

The clinical and microbiologic characteristics of 34 patients with extended-spectrum β-lactamase (ESBL) positive E. coli isolated from blood were compared to 66 bacteremic patients with ESBL negative E. coli, from January 2007 through December 2009. Of the 21 ESBL positive isolates available for PCR analysis, 13 were positive for CTX-M, 8 for TEM, 4 for SHV β-lactamases, with 6 possessing multiple enzymes. Twenty of 34 (59%) ESBL-positive and 41 of 66 (62%) ESBL-negative blood isolates were considered community-associated. All but one isolate in both groups had MICs of ≤1.0 μg/ml to meropenem. However, when compared to ESBL-negative isolates, ESBL-positive isolates were more frequently resistant to levofloxacin, trimethoprim/sulfamethoxazole and had higher MICs to gentamicin, tobramycin and piperacillin/tazobactam. The use of intravenous and urinary catheters was strongly associated with the isolation of E. coli bloodstream isolates in both groups of patients. Although hospital stay was similar in both groups, appropriate therapy was given in 87% of patients with ESBL positive vs. 98% of patients with ESBL negative isolates and mortality was greater for patients with ESBL positive isolates (26% vs. 17%). Since a large proportion of E. coli blood isolates were ESBL-positive and community-associated, carbapenems should be considered as initial empiric therapy for such infections in our locale.

Occurrence of Extended-Spectrum β-1actamase Producing Enterobacteriaceae （ESBLPE） among Primary School Pupil in Obafemi-Owode, Nigeria

Occurrence of extended-β-1actamase producing enterobacteriaceae （ESBLPE）, which has reduced the antibacterial efficacy and potency of many 3rd generation cephalosporins, was investigated among the primary school pupils. 88 primary school pupils in Obafemi-Owode Local Government, Southwestern Nigeria, including 49 males （55.7%） and 39 females （44,3%） （mean age 12 ± 3） were screened for ESBLPE isolates with exclusion criterion of antimicrobial use in the preceding 2 weeks either as therapy for gastro-intestinal complication or prophylaxis. ESBLPE detected include 4.5% of Eschericia coli, 2.3% of Enterobacter cloaca, 0% Proteus mirabilis, 2.3% Pseudomonas aeruginosa, 1.1% Staphylococcus aureus and 4.5% of Klebsiella oxytoca. 10 （76.9%） of ESBLPE isolates were resistant to disc of cefuroxime （30 μg）, 8 （61.5%） susceptible to amoxicillin/clavulanic （20/10 lag） and low susceptibility of 7 （53.8%） was recorded for ceftazidime （30 lag）. 0% susceptibility was recorded for the ESBLPE isolates to cefuroxime MIC 〉 8 gg/mL and ampicillin MIC 〉 8 lag/mL while E, coli and E. cloca each show 50.0% and P. aeruginosa and K. oxytoca show 100.0% and 75.0% susceptibility to augmentin （MIC 〈 8）. This study has shown a 14.7% proportion of the pupil to harbour ESBLPE from enteric source with increased resistant to most new generation cefuroximes. Therefore, transfer of virulent and antibiotic resistant ESBLPE could be aided by sharing feeding materials while fecal-oral route of transmission cannot be ruled out as hygiene level is very low thereby increasing emergence of virulent resistant enteric strains leading to treatment failure.

Phenotypic and Genotypic Characterization of Metallo-Beta-Lactamase and Extended-Spectrum Beta-Lactamase among Enterobacteria Isolated at National Public Health Laboratory of Brazzaville

The improper use of antimicrobials against infectious diseases has allowed microorganisms to develop defense mechanisms that give them insensitivity to these agents. All bacteria are concerned by this phenomenon. This work aimed to assess prevalence of beta-lactamase produced by enterobacterial isolates. Then, disc diffusion, double disc synergy test (DDST) and combined disc test (CDT) were respectively used for antimicrobial resistance, detection of Extended-Spectrum Beta-Lactamases (ESBL) and Metallo-Beta-Lactamases (MBL). bla genes were detected by PCR. A total of 132 enterobacterial strains were studied. Resistance to antibiotic families was observed with a greater frequency than 50%. Gentamicin was the least active beta-lactam antibiotic, with a resistance rate of 88%. 40.9% of strains show an ESBL phenotype and 16.6% were MBL. An overall prevalence of 74% (40/54) and respectively rates of 29.6%, 27.7% and 16.7% for blaSHV, blaCTX and blaTEM genes were observed. SHV, CTX, CTX/SHV/TEM, CTX/TEM, SHV/TEM and CTX/SHV were different ESBL genotypes observed. ESBL-producing enterobacteria isolation worried about the future of antimicrobial therapy in the Republic of Congo. This is a public health problem that requires careful monitoring and implementation of a policy of rational antibiotics use.

Unravelling Antimicrobial Resistance Phenotypes and Carriage of Extended-Spectrum β-Lactamase Genes in Escherichia coli Isolated from Dairy Farms in Kiambu County, Kenya

The use of antibiotics for prophylaxis and growth enhancement in livestock farming is on the increase globally. This practice has led to the emergence and spread of antimicrobial-resistant bacteria in livestock. Only limited research has been done to establish the role of cattle farming in antimicrobial resistance. The current study sought to establish the carriage of multi-drug resistance and extended-spectrum beta-lactamase genes in Escherichia coli from farmers, their cattle, and cattle slurry within Kiambu County. A total of 286 (81%) E. coli isolates were recovered from 352 samples analysed. Antibiotic resistance profiles showed 114 (40%) isolates were resistant to ≥3 antimicrobial classes and were considered multidrug-resistant. Among multidrug-resistant (MDR) E. coli strains, 40 (14%) were resistant to 3 different antimicrobial classes, while 71 (25%) were resistant to between 4 and 7 antibiotic classes. Extended-spectrum β-lactamase resistance was found in 18 isolates: human (n = 14), cattle (n = 2), and environmental (n = 2). Both the blaCTX-M and blaTEM genes were detected in 10 and 15 strains, respectively. Sequence analysis showed that the isolates carried the blaTEM-116 (n = 7), blaTEM-1 (n = 5), and blaCTX-M-15 (n = 8) genes. Genotyping MDR isolates using (GTG) 5 PCR demonstrated that the isolates were not clonal. This data shows antimicrobial resistance profiles and different types of resistance genes in the E. coli population on dairy farms. As a result, more effective, targeted public health policies and measures need to be put in place to control and prevent the emergence and spread of resistant bacteria.

Activity of Fosfomycin in Extended-Spectrum Beta-Lactamases Producing Klebsiella pneumonae from Hospital Acquired Urinary Tract Infections

Treatment of hospital acquired urinary tract infections (UTIs) caused by extended-spectrum beta-Lactamases producing Klebsiella pneumonae is a major problem. This organism expresses a high level of resistance to many groups of antibiotics. Fosfomycin is an agent which is recommended for treatment of UTIs caused by ESBLs producers. The aim of this study is to determine the sensitivity pattern of ESBLs producing urinary K. pneumonae to antimicrobial agents including fosfomycin in patients of MUHs and determine the prevalence of fosfomycin resistance mediated by plasmid mediated fosfomycin modifying enzymes fosA, fosB and fosA3. Methods: Klebsiella pneumonae urinary isolates were collected from patients with hospital acquired UTIs in Mansoura University Hospitals (MUHs). The susceptibility pattern was determined by Kirby Baur method. Isolates resistant to extended spectrum cephalosporins were tested for ESBLs production by double disc diffusion method. Fosfomycin resistance was determined by broth dilution method. Isolates resistant to fosfomycin were tested for fosA, fosB and fosA3 by PCR. Results: A total of 128 ESBLs producing K. pneumonae isolates were collected. The highest sensitivity was to imipenem (94.5%). The lowest was to trimethoprime-sulphamethoxazole (21.8%). Co-resistance of ESBLs isolates with fosfomycin was 23.2%. Eighteen fosfomycin resistant isolates (18/30) were positive to fosA. Conclusion: ESBLs producing urinary Klebsiella pneumonae express moderate sensitivity to fosfomycin. Resistance is mainly mediated by plasmid mediated fosfomycin modifying enzymes fosA.

Study on the Genotype of Extended-spectrum Beta-lactamases Mediated by Plasmid in Southern China

To investigate the prevalence and genotype of extended spectrum beta-lactamases (ESBLs) mediated by plasmid in Gram-negative bacteria found in southern China, a total of 1184 clinical isolates of non-repetitive strains of Gram-negative bacteria were collected in 2001 from 5 different cities in southern China. The ESBLs-producing isolates were distinguished by means of the phenotype confirmatory test based on the NCCLS criteria and were subjected to plasmid conjugation and electroporation experiments. Those clinical isolates succeeded in plasmid transfers had undergone plasmid conjugation and electro-transformation, plasmid DNA extraction and PstⅠ digest finger-printing analysis, as well as the universal primer PCR amplification of the TEM, SHV, CTX-M, VEB, PER and SFO genes and the DNA sequencing in order to determine the genotypes of ESBLs and their plasmid locations. It was found that the incidence of the ESBLs-producing strains of Gram-negative bacteria was 14.6% (173/1184) with 67 strains of transconjugants and 11 strains of electro-transformants, in which CTX-M-14 type was 33.3% (26/78); CTX-M-3 type was 23.1% (18/78); CTX-M-9 type was 14.1% (11/78); CTX-M-5 type was 6.4% ( 5/78); CTX-M-13 type was 2.6% (2/78); SHV-5 type was 7.7% (6/78); SHV-12 type was 5.1% (4/78), SHV-2a type was 2.6% (2/78) and unidentified type was 5.1% (4/78). 29.5% of the wild strains also carried broad-spectrum beta-lactamases TEM-1 and SHV-1 types. The above mentioned ESBLs genes were located on transferable plasmids with variable sizes (from 35 to 190?kb). The CTX-M type ESBLs was characterized by high-level of resistance to cefotaxime. It concluded that the CTX-M-type was the most prevalent genotype in clinical isolates of Gram-negative bacteria in southern China, and the SHV-type ranks in the second place. TEM-, VEB-, Toho- and PER-types were not found in these isolates.

Molecular Detection of Extended-Spectrum Beta Lactamase and Carbapenemase Genes in Escherichia coli and Salmonella spp. Isolated from Chickens Consumed Outdoors in Ouagadougou, Burkina Faso

This study was carried out to assess the prevalence of resistance genes in strains of Escherichia coli and Salmonella spp. isolated from free-range chickens in Ouagadougou, where resistant bacteria can be transmitted to humans via faeces or contaminated meat. A total of 280 strains of Escherichia coli and 129 strains of Salmonella spp. resistant to at least one beta-lactam or carbapenem antibiotic were used in this study. PCR analyses revealed the presence of ESBL (extended spectrum beta lactamase) resistance genes in Escherichia coli isolates, with 3.21% (9/280) possessing the CTX-M (Cefotaximase) gene, 15.35% (43/280) had the SHV (Sulfhydril Variable) gene, and 11.42% (32/280) had carbapenemase resistance genes, more specifically IMP (Imipenemase metallo-beta-lactamase). As regards Salmonella spp. strains, only the presence of the SHV (Sulfhydril Variable) gene was identified in 2.32% (3/129) of isolates belonging to the ESBL family, while 26.35% (34/129) and 13.95% (18/129) of isolates respectively possessed the IMP (Imipenemase metallo-beta-lactamase) and NDM (New Delhi metallo-β-lactamase) genes, both of the carbapenemase type. The significant prevalence of resistance genes in bacterial strains isolated from chickens sold outdoors in Ouagadougou raises major public health concerns, due to the possible transmission of these resistant strains to humans through the consumption of contaminated meat, thus complicating the treatment of bacterial infections.

Impact of Genetic Diversity of Uropathogenic Escherichia coli and Klebsiella pneumoniae Strains on the Dissemination of Extended Spectrum Beta-Lactam Resistance Genes in Côte d’Ivoire

The increase and spread of bacterial resistance to extended-spectrum beta-lactam antibiotics are reported in many infections and are a real public health problem worldwide. Drug pressure is a factor that favors the emergence of a population of better adapted bacteria. However, there is no literature highlighting the genetic diversity and evolutionary structure of E. coli and K. pneumoniae in an environment with high selection pressure in Côte d’Ivoire. The objective of this study was to evaluate the genetic diversity of E. coli and K. pneumoniae strains circulating at the HKB Hospital in Abobo and at the Daloa Regional Hospital and its impact on the dissemination of extended spectrum beta-lactam resistance genes. A total of 39 strains isolated from the urinary tract of infected patients, including 30 strains of E. coli and 9 strains of K. pneumoniae were studied. A total of 39 strains isolated from the urinary tract of infected patients, including 30 strains of E. coli and 9 strains of K. pneumoniae were studied. From genomic DNA extracts, ESBL resistance genes were amplified by PCR and sequenced, in addition to genetic typing by ERIC-PCR. The data obtained were submitted to genetic and bioinformatics analyses. The results have shown a genetic diversity important in E. coli and K. pneumoniae with diversity indexs (SID) ranging from 0.5 to 0.77. The genetic structure of the bacterial species studied has shown a clonal distribution of strains with clones expressing TEM-9 and CTX-M-15 variants. Also, this clonal structure was correlated with the spread of resistance genes in E. coli and K. pneumoniae. The spread of resistant clones is a factor that might limit the fight against antibiotic resistance.

Identification of Klebsiella pneumoniae strains harboring inactive extended-spectrum beta-lactamase antibiotic-resistance genes

Background The extended-spectrum beta-lactamase （ESBL）-producing Klebsiella pneumoniae has increasingly become a major contributor to nosocomial infections and can exhibit multiple antibiotic resistance.Previous studies have focused on the resistance genes in ESBL-producing strains,and the resistance-associated genetic environment of non-ESBL-producing strains has been ignored until now.Here,we investigated the occurrence and characteristics of non-ESBL-producing K.pneumoniae,which potentially carries unexpressed resistance genes.Methods K.pneumoniae strains were collected from five medical institutions in China from February 2010 to August 2013.The VITEK-2 ESBL detection system was used as a primary screen to identify the ESBL-producing phenotype,and the three primary types of ESBL-associated genes （CTX,SHV,and TEM） were detected by polymerase chain reaction （PCR） to confirm the strains presenting with a non-ESBL-producing phenotype.mRNA expression in the non-ESBL-producing strains was further screened by reverse-transcription PCR （RT-PCR） to validate their transcriptional efficiency.Results Out of 224 clinically isolated antibiotic-sensitive K.pneumoniae strains with a non-ESBL-producing phenotype,5 （2.2％） were identified to carry inactivated ESBL blaSHV genes with intact upstream promoter regions and resistance gene sequences.Interestingly,three of the five antibiotic-sensitive K.pneumoniae strains containing ESBL blaSHV genes still exhibited mRNA transcription of blasHv,while the other two exhibited no mRNA transcription.Conclusion These findings suggest that inactivated ESBL genes exist in non-ESBL-producing antibiotic-sensitive K.pneumoniae strains,which have the potential to transform the strain into an ESBL phenotype if an inappropriate application or overdose of antibiotics is implemented during clinical management.

In vitro study on the transmission of multidrug-resistant bacteria from textiles to pig skin

BACKGROUND The survival of microorganisms on textiles and specifically on healthcare profes-sionals’(HCP)attire has been demonstrated in several studies.The ability of microorganisms to adhere and remain on textiles for up to hours or days raises questions as to their possible role in transmission from textile to skin via HCP to patients.AIM To evaluate the presence,survival and transmission of different multidrug-resistant bacteria(MDRB)from HCP attire onto skin.METHODS Three MDRB[methicillin-resistant Staphylococcus aureus(MRSA);vancomycin-resistant Enterococcus faecium(VRE);carbapenem-resistant Klebsiella pneumoniae,(CRKP)]were inoculated on textiles from scrubs(60%cotton-40%polyester)and white coat(100%cotton)at concentrations of 108 colony-forming units(CFU),105 CFU,and 103 CFU per mL.The inoculation of swatches was divided in time intervals of 1 min,5 min,15 min,30 min,1 h,2 h,3 h,4 h,5 h,and 6 h.At the end of each period,textiles were imprinted onto pig skins and each skin square was inverted onto three different selective chromogenic media.Growth from the pig skin squares was recorded for the 3 MDRB at the three above concentrations,for the whole length of the 6-h experiment.RESULTS MRSA was recovered from pig skins at all concentrations for the whole duration of the 6-h study.VRE was recovered from the concentration of 108 CFU/mL for 6 h and from 105 CFU/mL for up to 3 h,while showing no growth at 103 CFU/mL.CRKP was recovered from 108 CFU/mL for 6 h,up to 30 min from 105 CFU/mL and for 1 min from the concentration of 103 CFU/mL.CONCLUSION Evidence from the current study shows that MRSA can persist on textiles and transmit to skin for 6 h even at low concentrations.The fact that all MDRB can be sustained and transferred to skin even at lower concentrations,supports that textiles are implicated as vectors of bacterial spread.

Genomic and Phenotypic Diversity of Carbapenemase-Producing Enterobacteriaceae Isolates from Bacteremia in China: A Multicenter Epidemiological, Microbiological, and Genetic Study

Carbapenemase-producing Enterobacteriaceae(CPE) isolates are recognized as one of the most severe threats to public health. However, the population structure and genetic characteristics of CPE isolates among bloodstream infections(BSIs) are largely unknown. To address this knowledge gap, in this study,we included patients with clinically significant BSIs due to Enterobacterales isolates, recruited from 26 sentinel hospitals in China(2014–2015). CPE isolates were microbiologically and genomically characterized,including their susceptibility profiles, molecular typing, phylogenetic features, and genetic context analysis of carbapenemase-encoding genes. Of the 2569 BSI Enterobacterales isolates enrolled, 42(1.6%) were carbapenemase-positive. Moreover, among the 2242 investigated isolates, 1111(49.6%) extendedspectrum β-lactamase(ESBL)-producing isolates were identified in Escherichia coli(E. coli), Klebsiella pneumoniae(K. pneumoniae), Proteus mirabilis(P. mirabilis), and Klebsiella oxytoca. Whole genome sequencing analysis showed the clonal spread of K. pneumoniae carbapenemase(KPC)-2-producing K. pneumoniae sequence type(ST) 11 and New Delhi metallo-β-lactamase(NDM)-5-producing E. coli ST167 in our collection. Plasmid analysis revealed that carbapenemase-encoding genes were located on multiple plasmids. A high prevalence of biofilm-encoding type 3 fimbriae clusters and yesiniabactin-associated genes was observed in K. pneumoniae isolates. This work demonstrates the high prevalence of ESBLs and the wide dissemination of CPE among BSI isolates in China, which represent real clinical threats. Moreover, our findings first illustrate a more comprehensive genome scenario of CPE isolates among BSIs. The clonal spread of KPC-2-producing K. pneumoniae ST11 and NDM-5-producing E. coli ST167 needs to be closely monitored.

Health care associated infections, antibiotic resistance and clinical outcome: A surveillance study from Sanandaj, Iran

AIM: To study the antibiotic susceptibility patterns of gram-negative healthcare associated bacterial infections at two tertiary hospitals in the Sanandaj city, Kurdistan Province, Iran.METHODS: From January 2012 to December 2012, all positive cultures from potentially sterile body fluids were gathered. They sent to professor Alborzi clinical microbiology center in Shiraz for further analysis and susceptibility testing. The antibiotic susceptibility was determined using the Kirby-Bauer method(disk diffusiontechnique). The Results were interpreted according to Clinical and Laboratory Standards Institute guidelines against a series of antimicrobials. World Health Organization definitions for Healthcare associated infections were followed.RESULTS: Seven hundred and thirty-two positive cultures were reported from both hospitals. Seventynine isolates/patients fulfilled the study criteria for healthcare associated gram-negative infections. The most frequent bacterial cultures were from the pediatric wards(52%). Serratia marcescens(S. marcescens)(38%) Escherichia coli(E. coli)(19%), Klebsiella pneumoniae(K. pneumoniae)(19%), Acinetobacter baumannii(6%), Enterobacter species(6%), Serratia odorifera(4%) and Pseudomonas species(5%) were the most frequently isolated organisms. The susceptibility pattern of common isolates i.e., S. marcescens, E. coli and K. pneumoniae for commonly used antibiotics were as follows: Ampicillin 3.3%, 6.7%, 20%; gentamicin 73.3%, 73.3%, 46.7%; ceftazidim 80%, 73.3%, 33.3%; cefepim 80%, 86.7%, 46.7%; piperacillin/tazobactam 90%, 66.7%, 86.7%; ciprofloxacin 100%, 73.3%, 86.7%; imipenem 100%, 100%, 100%, respectively. CONCLUSION: The most effective antibiotics against gram-negative healthcare associated infections are imipenem followed by ciprofloxacin. The resistance rate is high against ampicillin and cephalothin. The high mortality rate(46.1%) associated with S. marcescens is alarming.

Effect of "Fuzheng Qingre Tonglin" prescription on rats with complicated urinary tract infection caused by ESBLs-producing Escherichia coli

Objective: To observe the effect of traditional Chinese medicine "Fuzheng Qingre Tonglin"prescription on rats with complicated urinary tract infection caused by extended-spectrumβ-lactamases (ESBLs)-producing Escherichia coli and its possible mechanism. Methods:Specific-pathogen-free female SD rats with 7-9 weeks old were randomly divided into model group, levofloxacin group, low-dose Fuzheng Qingre Tonglin group, high-dose Fuzheng Qingre Tonglin group, levofloxacin+Fuzheng Qingre Tonglin group (n=8 in each above group), and also there was a control group with 5 rats. Each group was administered intragastrically 2 days after model establishment for 14 consecutive days. The general condition of the rats was observed. Renal pathological changes were observed by light microscopy under hematoxylin-eosin staining. The number of white blood cells and the percentage of neutral particles (NEUT%) were detected. ELISA was used for the determination of C-reactive protein (CRP), CD4+, CD8+ in the blood, and secretory immunoglobulin A (SIgA), N-acetyl-β-D-glucosidase (NAG) molecules in urine. Results: There were a large number of inflammatory cells in the kidney of the model group and the levofloxacin group. The inflammatory cells in the treatment group were significantly reduced. The NEUT% and CRP in the blood of each treatment group and the NAG in the urine were significantly lower than those in the model group (P<0.05), except levofloxacin group. Compared with the model group, except for the levofloxacin group, serum CD4 concentration and CD4/CD8 ratio in the other treatment groups were significantly decreased, and CD8 molecules were significantly increased (P<0.05). Conclusion: "Fuzheng Qingre Tonglin" can alleviate systemic and local inflammation of complicated urinary tract infections in rats. It may also have the effect of re-sensing the insensitive antibacterial drug to bacteria and may have the function of regulating immune function and protecting kidney function in rats.