Prevalence and Molecular Characterization of Fluoroquinolone-Resistant Escherichia coli in Healthy Children

Faecal E. coli can act as reservoirs for resistance genes. Here, we analyzed prevalence of drug resistance in faecal E. coli isolated from healthy children at a single kindergarten in Beijing, China, then used whole genome sequencing to characterize fluoroquinolone-non-susceptible strains. Our results revealed high resistance to ampicillin (54.0%), trimethoprim/sulphurmethoxazole (47.5%) and tetracycline (58.9%) among 576 faecal E. coli isolates, 49.2% of which exhibited multidrug resistance. A total of 113 E. coli isolates were not susceptible to ciprofloxacin, with four sequence types, namely ST1193 (25.7%), ST773 (13.3%), ST648 (8.8%) and ST131 (7.1%) found to be the most prevalent (54.9%). With regards to resistance to quinolones, we detected chromosomal mutations in gyrA, parC, and parE in 111 (98.2%), 105 (92.9%), and 67 (61.1%) isolates, respectively. blaCTX-M (37.2%) was the major ESBL gene, whereas blaCTX-M-14 (12.4%) and blaCTX-M-27 (11.5%) were the most frequent subtypes. A total of 90 (79.6%) ExPEC and 65 (57.5%) UPEC isolates were classified. Overall, these findings revealed clonal spread of certain prevalent STs, namely ST1193, ST773, ST648 and ST131 E. coli isolates in healthy children within a single kindergarten in Beijing, China, affirming the seriousness of the multidrug resistance problem and potential pathogenicity of E. coli isolates in healthy children. Therefore, there is an urgent need for increased surveillance to enhance control of this problem.

The Carrier Rate of Extended Spectrum Beta Lactamase (ESBL) Producing Bacteria in Cockroaches (Periplaneta americana) in Hospital and Community

Highlight :Bacteriologically for colonization of ESBL producing Enterobacteriaceae in cockroaches (Periplaneta americana) were analyzed.The prevalence of ESBL producing bacteria among cockroaches in hospitals is bigger than in households.Abstract: Cockroach (Periplaneta americana) is one of the vectors in the environment that can transmit disease. Cockroaches can act as potential mechanical vectors of antibiotic resistant bacteria. Enterobacteriaceae is a gram-negative bacteria that has natural habitats in the digestive tract of humans and animals. Enterobacteriaceae that produce Extended Spectrum β-lactamases (ESBLs) have emerged as major pathogens in hospitals. The study analyzed the prevalence of ESBL producing bacteria in cockroaches that lived in hospitals and residential homes. In this study, a total of 200 cockroaches consisting of 100 cockroaches from the hospital environment and 100 cockroaches from the residential environment were analyzed bacteriologically for colonization of ESBL producing Enterobacteriaceae. The specimen of the alimentary tract was taken and sub-cultured in MacConkey agar supplemented with cefotaxime 2 ug/ml. Growth colonies were suggested as an ESBL-producing bacteria, then were confirmed as ESBL producers by the Double Disk Synergy Test (DDST). The ESBL gene was detected by Polymerase Chain Reaction (PCR). Among 100 household cockroach samples, 14 (14%) were identified as ESBL producers, while 100 hospital cockroaches were 26 (26%) positive ESBL. The ESBL gene, in hospital cockroach were identified of CTXM 19 (19%), SHV 7 (7%), and not any TEM gene, while among household cockroaches were identified CTXM 2 (2%), SHV 11 (11%), and also not detected TEM ESBL gene. Among ESBL genes, only the CTXM gene was significantly different between household and hospital cockroaches.

ESBL/pAmpC-producing Escherichia coli and Klebsiella pneumoniae carriage among veterinary healthcare workers in the Netherlands

Background Animals are a reservoir for ESBL/pAmpC-producing Escherichia coli/Klebsiella pneumoniae (ESBL-E/K). We investigated the association between occupational contact with different types of animals and the prevalence of ESBL-E/K carriage among veterinary healthcare workers, assessed molecular characteristics of ESBL-E/K, and followed-up on the ESBL-E/K carriage status of participants and their household members. Methods Participants completed a questionnaire about their contact with animals at work and at home, health status, travel behaviour and hygiene, and sent in a faecal sample which was tested for the presence of ESBL-E/K. Resistance genes were typed using PCR and sequencing. ESBL-E/K positive participants and their household members were followed up after 6 months. Risk factors were analysed using multivariable logistic regression methods. Results The prevalence of ESBL-E/K carriage was 9.8% (47/482; 95%CI 7.4–12.7). The most frequently occurring ESBL genes were blaCTX-M-15, blaCTX-M-14 and blaDHA-1. The predominant sequence type was ST131. None of the occupation related factors, such as contact with specific animal species, were significantly associated with ESBL-E/K carriage, whereas travel to Africa, Asia or Latin America in the past 6 months (OR 4.4), and stomach/bowel complaints in the past 4 weeks (OR 2.2) were. Sixteen of 33 initially ESBL-E/K positive participants (48.5%) tested positive again 6 months later, in 14 persons the same ESBL gene and E. coli ST was found. Four of 23 (17.4%) household members carried ESBL-E/K, in three persons this was the same ESBL gene and E. coli ST as in the veterinary healthcare worker. Conclusions Despite the absence of specific occupation related risk factors, ESBL-E/K carriage in veterinary healthcare workers was high compared to the prevalence in the general Dutch population (5%). This indicates that occupational contact with animals is a potential source of ESBL-E/K for the population at large.

Circulation of third-generation cephalosporin resistant Salmonella Typhi in Mumbai, India

We report the persistent circulation of third-generation cephalosporin resistant Salmonella Typhi in Mumbai, linked to the acquisition and maintenance of a previously characterized IncX3 plasmid carrying the ESBL gene blaSHV-12 and the fluoroquinolone resistance gene qnrB7 in the genetic context of a triple mutant also associated with fluoroquinolone resistance.

Occurrence of Escherichia coli producing extended spectrum β-lactamases in food-producing animals

Multidrug resistance due to the production of extended-spectrum beta-lactamases (ESBLs) is a major problem in human as well as in veterinary medicine. These strains appear in animal and human microbiomes and can be the source of infection both in animal and in human healthcare, in accordance with the One Health theorem. In this study we examined the prevalence of ESBL-producing bacteria in food-producing animals. We collected 100 porcine and 114 poultry samples to examine the prevalence of ESBL producers. Isolates were identified using the MALDI-TOF system and their antibiotic susceptibility was tested using the disk diffusion method. ESBL gene families and phylogroups were detected by polymerase chain reactions. The prevalence of ESBL producers was relatively high in both sample groups: 72 (72.0%) porcine and 39 (34.2%) poultry isolates were ESBL producers. Escherichia coli isolates were chosen for further investigations. The most common ESBL gene was CTX-M-1 (79.3%). Most of the isolates belong to the commensal E. coli phylogroups. The porcine isolates could be divided into three phylogroups, while the distribution of the poultry isolates was more varied. In summary, ESBL-producing bacteria are prevalent in the faecal samples of the examined food-producing animals, with a dominance of the CTX-M-1 group enzymes and commensal E. coli phylogroups.

Circulation of third-generation cephalosporin resistant Salmonella Typhi in Mumbai, India

We report the persistent circulation of third-generation cephalosporin resistant Salmonella Typhi in Mumbai, linked to the acquisition and maintenance of a previously characterized IncX3 plasmid carrying the ESBL gene blaSHV-12 and the fluoroquinolone resistance gene qnrB7 in the genetic context of a triple mutant also associated with fluoroquinolone resistance.

From the Urinary Catheter to the Prevalence of Three Classes of Integrons, β-Lactamase Genes, and Differences in Antimicrobial Susceptibility of Proteus mirabilis and Clonal Relatedness with Rep-PCR

Introduction. Proteus mirabilis is a biofilm-forming agent that quickly settles on the urinary catheters and causing catheter-associated urinary tract infections. Thus, the spread of multidrug-resistant P. mirabilis isolates, with the ability to form a biofilm that carries integron, extended-spectrum β-lactamases (ESBLs), and plasmid-mediated colistin resistance genes (mcr), represents a severe threat to managing nosocomial infectious diseases. This study is aimed at surveying the prevalence of ESBL, integrase, and mcr genes of P. mirabilis, isolated from the catheter, to assess the differences in their antimicrobial susceptibility and clonal dissemination. Method. Microtiter plate assay was adopted to measure biofilm formation. The antimicrobial susceptibility was assessed by the disk diffusion method. Antimicrobial resistance genes (intI1, intI2, intI3, blaTEM, blaCTX-M, blaSHV, mcr1, and mcr2) were detected by PCR. All of the isolates were characterized by repetitive sequence-based PCR. Result. From 385 collected catheters in patients admitted to the intensive care unit (ICU), 40 P. mirabilis were isolated. All of the isolates could form a biofilm. Proteus spp. had intrinsic resistance to tetracycline (95%) and nitrofurantoin (92.5%), which explains the high resistance prevalence. The most widely resistant antibiotic was trimethoprim-sulfamethoxazole (75%). Thirty-three (82.5%) isolates were classified as multidrug resistance (MDR). The prevalence of intI1 and intI2 genes was 60% and 25%, respectively. In 6 (15%) isolates, both genes were detected. The most frequent ESBL gene detected in all of the isolates was blaTEM. Also, no detection for mcr1 and mcr2 antibiotic resistance genes was reported. Rep-PCR identified 39(GTG)5 types (G1–G39) of 40 isolates that 38 isolates had unique patterns. Conclusion. In this study, 82.5% of isolates were MDR with high antibiotic resistance to trimethoprim-sulfamethoxazole. The intI1 and blaTEM were the most prevalent genes in the integrase and ESBL gene family. High diversity was seen in the isolates with Rep-PCR. The increasing rate of MDR isolates with a high prevalence of resistance genes could be alarming and demonstrate the need for hygienic procedures to prevent the increased antibiotic resistance rate in the future.

Existence of Multiple ESBL Genes among Phenotypically Confirmed ESBL Producing Klebsiella pneumoniae and Escherichia coli Concurrently Isolated from Clinical, Colonization and Contamination Samples from Neonatal Units at Bugando Medical Center, Mwanza, Tanzania

The proportions and similarities of extended-spectrum β-lactamase (ESBL) producing K. pneumoniae (ESBL-KP) and E. coli (ESBL-EC) carrying multiple ESBL genes is poorly known at our setting. This study investigated the existence of multiple ESBL genes (blaCTX-M, blaTEM, and blaSHV) among ESBL-KP and ESBL-EC concurrently isolated from clinical, colonization, and contamination samples from neonatology units in Mwanza-Tanzania. Twenty and 55 presumptive ESBL-EC and ESBL-KP, respectively, from a previous study archived at −80 °C were successfully recovered for this study. Isolates were screened and confirmed for production of ESBLs by phenotypic methods followed by multiplex PCR assay to determine ESBL genes. All (100%) and 97.3% of presumptive ESBL isolates were phenotypically confirmed by Clinical and Laboratory Standards Institute (CLSI) and modified double-disc synergy methods, respectively. About 93.3% (70/75) of phenotypically confirmed ESBL isolates had at least one ESBL gene, whereby for 62.9% (44/70), all ESBL genes (blaCTX-M, blaTEM, and blaSHV) were detected. Eight pairs of ESBL bacteria show similar patterns of antibiotics susceptibility and ESBL genes. ESBL-KP and ESBL-EC, concurrently isolated from clinical, colonization and contamination samples, harbored multiple ESBL genes. Further, eight pairs of ESBL isolates had similar patterns of antibiotics susceptibility and ESBL genes, suggesting transmission of and/or sharing of mobile genetic elements (MGEs) among ESBL-KP and ESBL-EC.

Draft Genome Sequences of Seven Extended-Spectrum β-Lactamase-Producing Escherichia coli Strains Isolated from New Zealand Waterways

ABSTRACT Draft genomes of seven extended-spectrum β-lactamase (ESBL)-producing Escherichia coli strains recovered from New Zealand waterways are described. The mean genome size was 5.1 Mb, with 4,724 coding sequences. All genomes contained the ESBL gene blaCTX-M, and one carried a plasmid-mediated AmpC gene, blaCMY-2. A multidrug-resistant genotype was detected in three isolates.

Prevalence of Carriage of Extended-Spectrum Beta-Lactamase-Producing Enterobacteriaceae Among Pregnant Women in the Primary Health Care Center and Hospital Setting in Indonesia

Background: The incidence of healthy individuals carrying Extended-Spectrum Beta-Lactamase producing Enterobacteriaceae (ESBL-E), especially E. coli (ESBL-EC) and Klebsiella pneumoniae (ESBL-KP), is increasing worldwide. ESBL-E causes early or late onset of neonatal sepsis, resulting in increased morbidity and mortality rates. Although maternal-neonatal transmissions of ESBL-E have been reported in several countries, the prevalence of ESBL-E carriage among pregnant women in Indonesia is not clear. In the present study, we compared the prevalence of carriage of ESBL-E among pregnant women in a primary health care center (PHC) versus two hospitals in Indonesia and identified the phenotypic and genotypic characteristics of the isolated ESBL-E strains. Methods: We collected rectal swab samples from 200 pregnant women who visited a PHC (101 women) or were admitted to Dr. Soetomo Referral Hospital or Airlangga University Hospital (99 women) in Surabaya, Indonesia from July to October 2018. The samples were cultivated on MacConkey Agar plates supplemented with cefotaxime 2 mg/L, at 37oC overnight. The isolated strains were identified by Bruker MALDI BiotyperÒ System, phenotypic detection of ESBL was performed by the combination disk method, and antibiotic susceptibility was tested by the disk diffusion method. In addition, ESBL gene was identified by PCR and DNA sequencing and molecular epidemiological studies were performed by PFGE. Results: ESBL-E strains were isolated from 25 (rate of fecal carriage; 24.8%) pregnant women who visited the PHC and 49 (49.5%) pregnant women who were admitted to the hospitals. The rate of ESBL-E carriage of pregnant women in the hospitals was significantly higher than that in the PHC. Among the 74 isolated ESBL-E strains, ESBL-EC was most frequently isolated (62 strains), followed by ESBL-KP (12 strains). In addition, blaCTX-M15 was the most frequent ESBL gene type of the isolated ESBL-E strains. Conclusion: Our results revealed the high prevalence of ESBL-E carriage in pregnant women, especially those who were admitted to the hospitals. CTX-M15 ESBL-EC was the most frequent type of ESBL-E in the pregnant women in our study. Continuous surveillance for ESBL-E carriage in pregnant women is strongly recommended to reduce the incidence of neonatal sepsis in Indonesia.