Multidrug-resistant Enterobacteriaceae Research Articles

Sewage from a secondary hospital in Ribeirão Preto, southeastern Brazil: a source of multidrug-resistant Enterobacteriaceae.

Antimicrobial resistance is one of the severe threats to global health. Hospital sewage can serve as a reservoir for multi-resistant bacteria and promote the spread of antimicrobial resistance. This study aimed to investigate the antimicrobial susceptibility and the pathogenic potential of Enterobacteriaceae isolated from the sewage of a secondary hospital in Ribeirão Preto, a city in southeastern Brazil. The strains were isolated by membrane filtration and identified by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF). The antimicrobial susceptibility profile was performed by disk diffusion. Polymerase chain reaction (PCR) assays were used to detect virulence genes among the strains. Twenty-eight isolates were obtained, with Klebsiella pneumoniae being the predominant species (71.4%, n = 20). All isolates were classified as multidrug-resistant, including four isolates that were non-susceptible to at least 50% of the tested antibiotics. All isolates were also non-susceptible to cefuroxime and sulfonamides antibiotics; however, they were susceptible to norfloxacin, ofloxacin, amikacin, gentamicin, netilmicin, ertapenem, cefazolin, cefaclor, and cefotetan. The virulence genes ycfM, fimH, mrkD, kfu, and entB were detected in several isolates. Our study showed that even in a secondary hospital, without the routine of major surgeries and intensive care admissions, the hospital sewage can harbor a high percentage of multidrug-resistant bacteria with pathogenic potential. This leads to the worrying risk of public health and environmental contamination.

Enhancement of anti-bactericidal and anti-biofilm activities of silver nanoparticles against multidrug-resistant enteric pathogens isolated from children with diarrhea

Background: Gastroenteritis has a significant mortality andmorbidity incidence in children globally. Antimicrobial resistance in Enterobacteriaceaeis a serious public health problem, especially in developing countries.Objective: The present study aimed to evaluate the anti-bacterial and anti-biofilmactivities of AgNPs alone and in combination with kanamycin against multidrug-resistant Enterobacteriaceae and P. aeruginosa isolated from diarrheal children.Materials and methods: 90 Enterobacteriaceae and P. aeruginosa isolates fromdiarrheal children were evaluated against 10 antibiotics. Minimum inhibitory doses ofAgNPs and kanamycin were determined using broth microdilution, synergistic wasdetermined using Checkerboard dilution tests, and the Calgary technique was used toanalyze biofilm development.Results: A total of 90 stool cultures were conducted for bacteria associated withdiarrhea among children attending some Baghdad hospitals. The findings revealed thatbacterial diarrhea was most often caused by E. coli 31 (34.5%), followed by S. typhi 19(21.1%), K. pneumoniae 14 (15.5%), P. aeruginosa 11 (12.2%) and S. sonnei 6 (6.7%),and significant variations between the strain's species were discovered using statisticalanalysis (P < 0.05). The present study's findings revealed that bacteria isolated fromchildren with diarrhea were spread significantly in age groups of 37–48 months andsignificantly different between age groups (P < 0.05), with a male/female ratio of0.57/1. Imipenem and amikacin were the most active antibiotics compared to penicillin,which was the least effective antibiotic. The combination of sublethal doses of AgNPswith sub-MIC (½MIC) of kanamycin exhibited substantial synergistic bactericidaleffects against MDR-Enterobacteriaceae. AgNPs inhibited biofilm formation by55%–65% for diarrhea-causing bacteria, while the combination of AgNPs withkanamycin demonstrated the strongest biofilm inhibition of around 80%–90% againstMDR-Enterobacteriaceae with a highly significant variation (P < 0.05).Conclusions: The outcomes of the research shows that the combination of AgNPs withkanamycin has remarkable synergistic bactericidal and anti-biofilm effectivenessagainst MDR-Enterobacteriaceae isolated from diarrheal children.

A novel cyclic dipeptide from Lactiplantibacillus plantarum MC39 inhibits proliferation of multidrug-resistant Klebsiella pneumoniae W8 and Enterobacter hormaechei U25

The present study pays special attention to lactic acid bacteria products, cyclic dipeptides (CDPs), as an effective prevention strategy that can be used to eradicate the threats posed by multidrug-resistant (MDR) Enterobacteriaceae. Herein, Lactiplantibacillus plantarum MC39, a strain isolated from an artisanal Egyptian cheese, was a producer of a novel CDP that could strongly inhibit, in vitro, MDR Klebsiella pneumoniae W8 and Enterobacter hormaechei U25, for the first time. Following partial purification with 80% ammonium sulphate precipitation and Sephadex G-100 chromatography, this biomolecule was identified as the cyclic dipeptide cyclo(Ala–Glu) using IR, HNMR, and LC–MS. Cyclo(Ala–Glu) was thermostable up to 121 °C, insensitive to pepsin and α-chymotrypsin, tolerant to organic solvents, active at acidic pH, and stable for 45 days of storage. Thus, it shows clear evidence to be usable as a bio-preservative in dairy products and as a potential therapeutic agent, controlling MDR K. pneumoniae and E. hormaechei.

Prevalence and Antimicrobial Susceptibility Pattern of Enterobacteriaceae Isolated from Hospitals Wastewater

Background: Hospital wastewater (HWW) is bearing in mind the main root to proliferate multidrug-resistant among the Enterobacteriaceae family. Enterobacteriaceae is suited to be a considerable risk for antibiotic-resistant through CTX-M production which is observed as an important factor leading to multiple drug-resistant. Study Design: A total of 100 samples of HWW and community wastewater were collected aseptically. Samples were sub-cultured through basic, selective, and UTI Chrome agar. Place of the study: This research was conducted in the Postgraduate Research Laboratory of the Department of Microbiology, Government College University Faisalabad. Duration of the study: The duration of the study was started from September 2018 to May 2019. Methodology: Bacteria were identified based on cultural characteristics and different biochemical tests. Antimicrobial susceptibility testing was performed as per CLSI 2016 guidelines. The molecular characterization of the CTX-M was done bt PCR to present its association with multidrug-resistant. Results: Among the total 165 isolates, the predominant bacteria were E.coli (48.1%) followed by K. pneumonia (25.3%), Shigella species (8.0%), and P.aeruginosa (12.7%) from hospital wastewater while E.coli (30.3%) followed by K. pneumonia (32.6%), Shigella species (20.9%) and P.aeruginosa (11.6%) were isolated from community wastewater. The AST of CTX-M producers and Non-CTX-M producers was also compared to find the relation of CTX-M to multidrug-resistant. Tigecycline was observed as a predominant resistant (≥96%) antibiotic followed by meropenem (≥40%) against all isolated CTX-M producers. Conclusion: The conclusion represented the existence of MDR Enterobacteriaceae in untreated hospital wastewater. All efforts revealed realize the requisite that hospital wastewater should be treated with standard treatment plants before exposing it to community wastewater. Keywords: Hospital wastewater; Antibiotic Resistance genes; CTX-M producers; Urinary Tract Infection (UTI),Enterobacteriaceae

Increasing trends of colistin resistance in patients at high-risk of carbapenem-resistant Enterobacteriaceae

Introduction Occurrence of colistin-resistant Enterobacteriaceae in response to the unregulated use of this antibiotic has been documented. This study reports an investigation of colistin resistance rates among carbapenem-resistant enterobacterial clinical isolates. Methods A total of 196 multidrug-resistant Enterobacteriaceae isolates (Klebsiella pneumoniae (n = 100), Escherichia coli (n = 89) and Enterobacter cloacae (n = 7) were selected from Gram-negative isolates over one year. Susceptibility to antimicrobials was determined using Vitek2. Broth microdilution method was used to detect colistin antimicrobial susceptibility. Identification of ESBL and carbapenemases were both done phenotypically and by PCR. Results All the studied isolates showed multidrug-resistant phenotypes with 51.5% resistance to carbapenems (meropenem, imipenem). Very low resistance rates towards tigecycline (n = 9) 4.6% were found. Thirty-nine isolates (19.9%) showed reduced susceptibility to colistin among the MDR isolates. Sixty-four isolates (32.7%) were ESBL producers. Hundred isolates (51%) were carbapenemase producers using Carba NP test. The PCR amplification results revealed that 40 isolates (20%) harboured NDM-1 and 40 isolates contained OXA-48-like gene. Coexistence of both (NDM-1 and OXA-48-like) was observed in nine (4.59%) isolates. A Statistically significant relationship was observed between carbapenem resistance and each of the followings; OXA-48 producers (p= .009), amikacin resistance (p = .000), gentamicin resistance (p = .032), tobramycin resistance (p = .000), and tigecycline resistance (p-value ≤ .001). A statistical significance was detected between ESBL-producing isolates and carbapenem susceptible isolates ESBL producers with p = 0.000. Conclusion An alarming sign is the increasing colistin resistance rates among carbapenem-resistant isolates. Aminoglycosides are still a therapeutic option to decrease the use of colistin and avoid further development of resistance. KEY MESSAGES High rates of colistin resistance among carbapenem-resistant Enterobacteriaceae. The choice of antibiotic is significantly associated with the clinical site of infection. Aminoglycosides are offered choices for treating multiple drug-resistant Enterobacteriaceae to preserve the colistin and carbapenems.

Invitro Activity of Tigecycline against Multidrug-Resistant Enterobacteriaceae from Blood Stream Infection in a Tertiary Care Hospital of Nepal

Background: Multidrug resistant (MDR) enterobacteriaceae like extended spectrum beta lactamase (ESBL) and metallo-beta lactamase (MBL) are being encountered as the causative agents of blood stream infection. To overcome this, accurate diagnosis of MDR pattern with appropriate antibiotics is required. Tigecycline is a broad spectrum antibiotic which exhibit strong activity against MDR enterobacteriaceae. This study is aimed to find out resistance pattern like ESBL and MBL with invitro activity of tigecycline against enterobacteriaceae. Methods: A descriptive cross-sectional study was conducted in the Department of Microbiology, B.P. Koirala Institute of Health Sciences, from 1st September 2014 to 31st August 2015. Confirmation for ESBL was done as recommended by Clinical and Laboratory Standard Institute (CLSI) and MBL production was detected by double disk synergy test. Antibiotic sensitivity test against tigecycline was done by Kirby-Bauer disk diffusion method. Results: 192 (1.70%) enterobacteriaceae were isolated throughout the study. Among them, 94 (49%) were ESBL, 51 (26.5%) were carbapenemase and 22 (11.5%) were MBL producers. A total of 64 (33.4%) isolates were found to be MDR. None of the isolates was resistant against tigecycline. Conclusion: Tigecycline is found to have excellent invitro activity against MDR enterobacteriaceae from BSI.

Detection of an IMI-2 carbapenemase-producing Enterobacter asburiae at a Swedish feed mill.

Occurrence of multidrug resistant Enterobacteriaceae in livestock is of concern as they can spread to humans. A potential introduction route for these bacteria to livestock could be animal feed. We therefore wanted to identify if Escherichia spp., Enterobacter spp., Klebsiella spp., or Raoutella spp. with transferable resistance to extended spectrum cephalosporins, carbapenems or colistin could be detected in the environment at feed mills in Sweden. A second aim was to compare detected isolates to previous described isolates from humans and animals in Sweden to establish relatedness which could indicate a potential transmission between sectors and feed mills as a source for antibiotic resistant bacteria. However, no isolates with transferable resistance to extended-cephalosporins or colistin could be identified, but one isolate belonging to the Enterobacter cloacae complex was shown to be carbapenem-resistant and showing carbapenemase-activity. Based on sequencing by both short-read Illumina and long-read Oxford Nanopore MinIon technologies it was shown that this isolate was an E. asburiae carrying a bla IMI-2 gene on a 216 Kbp plasmid, designated pSB89A/IMI-2, and contained the plasmid replicons IncFII, IncFIB, and a third replicon showing highest similarity to the IncFII(Yp). In addition, the plasmid contained genes for various functions such as plasmid segregation and stability, plasmid transfer and arsenical transport, but no additional antibiotic resistance genes. This isolate and the pSB89A/IMI-2 was compared to three human clinical isolates positive for bla IMI-2 available from the Swedish antibiotic monitoring program Swedres. It was shown that one of the human isolates carried a plasmid similar with regards to gene content to the pSB89A/IMI-2 except for the plasmid transfer system, but that the order of genes was different. The pSB89A/IMI-2 did however share the same transfer system as the bla IMI-2 carrying plasmids from the other two human isolates. The pSB89A/IMI-2 was also compared to previously published plasmids carrying bla IMI-2, but no identical plasmids could be identified. However, most shared part of the plasmid transfer system and DNA replication genes, and the bla IMI-2 gene was located next the transcription regulator imiR. The IS3-family insertion element downstream of imiR in the pSB89A was also related to the IS elements in other bla IMI-carrying plasmids.

Lactobacillus supports Clostridiales to restrict gut colonization by multidrug-resistant Enterobacteriaceae

Infections by multidrug-resistant Enterobacteriaceae (MRE) are life-threatening to patients. The intestinal microbiome protects against MRE colonization, but antibiotics cause collateral damage to commensals and open the way to colonization and subsequent infection. Despite the significance of this problem, the specific commensals and mechanisms that restrict MRE colonization remain largely unknown. Here, by performing a multi-omic prospective study of hospitalized patients combined with mice experiments, we find that Lactobacillus is key, though not sufficient, to restrict MRE gut colonization. Lactobacillus rhamnosus and murinus increase the levels of Clostridiales bacteria, which induces a hostile environment for MRE growth through increased butyrate levels and reduced nutrient sources. This mechanism of colonization resistance, an interaction between Lactobacillus spp. and Clostridiales involving cooperation between microbiota members, is conserved in mice and patients. These results stress the importance of exploiting microbiome interactions for developing effective probiotics that prevent infections in hospitalized patients.

The effect of re-directed patient flow in combination with targeted infection control measures on the spread of multi-drug-resistant Enterobacteriaceae in the German health-care system: a mathematical modelling approach

ObjectiveThe introduction of multi-drug-resistant Enterobacteriaceae (MDR-E) by colonized patients transferred from high-prevalence countries has led to several large outbreaks of MDR-E in low-prevalence countries, with the risk of propagated spread to the community. The goal of this study was to derive a strategy to counteract the spread of MDR-E at the regional health-care network level. MethodsWe used a hybrid ordinary differential equation and network model built based on German health insurance data to evaluate whether the re-direction of patient flow in combination with targeted infection control measures can counteract the spread of MDR-E in the German health-care system. We applied pragmatic re-direction strategies focusing on a reduced choice of hospitals for subsequent stays after initial hospitalization but not manipulating direct transfers because these are most likely determined by medical needs. ResultsThe re-direction strategies alone did not reduce the system-wide spread of MDR-E (system-wide prevalence of MDR-E is 18.7% vs. 25.7%/29.9%). In contrast, targeted hospital-based infection control measures restricted to institutions with the highest institutional basic reproduction numbers in the network were identified as an effective tool for reducing system-wide prevalence (system-wide prevalence of MDR-E is 18.7% vs. 9.3%). If these measures were applied to the top one-third of hospitals, the system-wide prevalence could be reduced by approximately 80% (system-wide prevalence of 18.7% vs. 3.5% for one-third of patients subjected to interventions). A combination of this hospital-based intervention and patient re-direction strategies could not improve the effectiveness of the hospital-based approach (system-wide prevalence of MDR-E is 9.3% vs. 14.2%/14.3%). ConclusionsThe pragmatic patient re-direction strategies were not capable of restricting the spread of MDR-E in a simulation of the German health-care system; in contrast, hospital-based interventions focusing on institutions identified based on network transmission patterns seem to be a promising approach for sustainable reduction of the spread of MDR-E through the German population.

The diversity in antimicrobial resistance of MDR Enterobacteriaceae among Chinese broiler and laying farms and two mcr-1 positive plasmids revealed their resistance-transmission risk.

This research aimed to investigate the microbial composition and diversity of antimicrobial resistance genes (ARGs) found in Chinese broiler and layer family poultry farms. We focused on the differences in resistance phenotypes and genotypes of multidrug-resistant Enterobacteriaceae (MDRE) isolated from the two farming environments and the existence and transmissibility of colistin resistance gene mcr-1. Metagenomic analysis showed that Firmicutes and Bacteroides were the dominant bacteria in broiler and layer farms. Many aminoglycoside and tetracycline resistance genes were accumulated in these environments, and their absolute abundance was higher in broiler than in layer farms. A total of 526 MDRE were isolated with a similar distribution in both farms. The results of the K-B test showed that the resistance rate to seven antimicrobials including polymyxin B and meropenem in broiler poultry farms was significantly higher than that in layer poultry farms (P ≤ 0.05). PCR screening results revealed that the detection rates of mcr-1, aph(3’)Ia, aadA2, blaoxa–1, blaCTX–M, fosB, qnrD, sul1, tetA, and catA1 in broiler source MDRE were significantly higher than those in layers (P ≤0.05). A chimeric plasmid p20432-mcr which carried the novel integron In1866 was isolated from broiler source MDRE. The high frequency of conjugation (10–1 to 10–3) and a wide range of hosts made p20432-mcr likely to play an essential role in the high detection rate of mcr-1, aph(3’)-Ia, and aadA2 in broiler farms. These findings will help optimize disinfection and improve antimicrobial-resistant bacteria surveillance programs in poultry farms, especially broilers.

PHENOTYPIC AND GENOTYPIC DETECTION OF CARBAPENAMASE PRODUCTION AMONG ENTEROBACTERIACEAE IN A TERTIARY CARE HOSPITAL, PORT BLAIR, INDIA

Background and Objective Multidrug resistance is emerging worldwide at an alarming rate among a variety of bacterial species, causing both community-acquired and nosocomial infections, including Enterobacteriaceae. The clinical utility of carbapenems, the agents of last resort against multi-drug resistant Enterobacteriaceae, is under threat with the growing incidence of pan resistant isolates. This study aims to determine the prevalence of carbapenemase production among Enterobacteriaceae, as such data is not available from this area. The study, prospective in design, was conducted during the period from April 2016 -Materials and Methods August 2018. Any isolate reistant to IMP and/or MRP were subjected to phenotypic detection (Modied Hodge Test, MHT and Double Disk Synergy Test, DDST) and genotypic (blaIMP, blaKPC, blaVIM, blaOXA-48, blaOXA-23, blaOXA-51, blaOXA-58 and blaNDM) basis of carbapenamase production. A total of 218 consecutive isolates of Enterobacteriaceae, which were resistant to IMP and/or MRP, wereResult: considered for the study. The most common isolates of Enterobacteriaceae were Klebsiella pneumoniae (n=122, 55.5%) followed by Escherichia coli (n=80, 36.4%). Modied Hodge Test (MHT) were positive for 12, 02 and Double Disc Synergy Test (DDST) were 11, 05 for Klebsiella and E.coli respectively. Multiplex PCR I and II, was performed for 36 isolates. The carbapenemase producing organisms were positive for 60% for single gene and 36% for multiple genes. Delay in detection of carbapenemase producing organisms result in longer hospitalizationConclusion: and increased health-care cost and this further affects the morbidity and mortality of patients in hospital set up.

Co-transfer of mcr-8 with blaNDM-1 or tmexCD1-toprJ1 by plasmid hybridisation

Carbapenems, tigecycline and colistin are three important antimicrobial agents for the treatment of clinical infections caused by multidrug-resistant Enterobacteriaceae. Here we characterised the formation of hybrid plasmids containing mcr-8 and blaNDM-1 or tmexCD1-toprJ1 that could confer resistance to colistin and carbapenems or tigecycline. More specifically, these clinically important genes could be co-transferred through IS26- and ltrA-mediated plasmid fusion to clinical isolates during conjugation under single drug (colistin) selection, following which the recipient strains became carbapenem- or tigecycline-resistant. The transferability and stability of these hybrid multidrug resistance (MDR) plasmids depend on the bacterial host and the presence of antibiotics. Further evolution and adaptation of these hybrid plasmids may facilitate their emergence and spread, which is of great concern for clinical therapy.

The use of high-affinity polyhistidine binders as masking probes for the selection of an NDM-1 specific aptamer

The emergence of carbapenemase-producing multi-drug resistant Enterobacteriaceae poses a dramatic, world-wide health risk. Limited treatment options and a lack of easy-to-use methods for the detection of infections with multi-drug resistant bacteria leave the health-care system with a fast-growing challenge. Aptamers are single stranded DNA or RNA molecules that bind to their targets with high affinity and specificity and can therefore serve as outstanding detection probes. However, an effective aptamer selection process is often hampered by non-specific binding. When selections are carried out against recombinant proteins, purification tags (e.g. polyhistidine) serve as attractive side targets, which may impede protein target binding. In this study, aptamer selection was carried out against N-terminally hexa-histidine tagged New Delhi metallo-ß-lactamase 1. After 14 selection rounds binding to polyhistidine was detected rather than to New Delhi metallo-ß-lactamase 1. Hence, the selection strategy was changed. As one aptamer candidate showed remarkable binding affinity to polyhistidine, it was used as a masking probe and selection was restarted from selection round 10. Finally, after three consecutive selection rounds, an aptamer with specific binding properties to New Delhi metallo-ß-lactamase 1 was identified. This aptamer may serve as a much-needed detection probe for New Delhi metallo-ß-lactamase 1 expressing Enterobacteriaceae.

Evaluating the Potential and Synergetic Effects of Microcins against Multidrug-Resistant Enterobacteriaceae.

ABSTRACTThe advent of multidrug-resistant bacteria has hampered the development of new antibiotics, exacerbating their morbidity and mortality. In this context, the gastrointestinal tract reveals a valuable source of novel antimicrobials. Microcins are bacteriocins produced by members of the family Enterobacteriaceae, which are endowed with a wide diversity of structures and mechanisms of action, and exert potent antibacterial activity against closely related bacteria. In this study, we investigated the antibacterial activities of four microcins against 54 Enterobacteriaceae isolates from three species (Escherichia coli, Klebsiella pneumoniae, and Salmonella enterica). The selected microcins, microcin C (McC, nucleotide peptide), microcin J25 (MccJ25, lasso peptide), microcin B17 (MccB17, linear azol(in)e-containing peptide), and microcin E492 (MccE492, siderophore peptide) carry different post-translational modifications and have distinct mechanisms of action. MICs and minimal bactericidal concentrations (MBC) of the microcins were measured and the efficacy of combinations of the microcins together or with antibiotics was assessed to identify potential synergies. Every isolate showed sensitivity to at least one microcin with MIC values ranging between 0.02 μM and 42.5 μM. Among the microcins tested, McC exhibited the broadest spectrum of inhibition with 46 strains inhibited, closely followed by MccE492 with 38 strains inhibited, while MccJ25 showed the highest activity. In general, microcin activity was observed to be independent of antibiotic resistance profile and strain genus. Of the 42 tested combinations, 20 provided enhanced activity (18 out of 20 being microcin–antibiotic combinations), with two being synergetic.IMPORTANCE With their wide range of structures and mechanisms of action, microcins are shown to exert antibacterial activities against Enterobacteriaceae resistant to antibiotics together with synergies with antibiotics and in particular colistin.

High Prevalence of Multi-Drug Resistance and Extended-Spectrum Beta-Lactamase-Producing Enterobacteriaceae Among Hospitalized Patients Presumptive for Bacterial Infection at Debre Berhan Comprehensive Specialized Hospital, Ethiopia.

BackgroundMulti-drug resistant Enterobacteriaceae (MDR-E), primarily extended-spectrum beta-lactamase producers (ESBLs), have emerged as a major public health concern. This study aimed to determine the prevalence of multi-drug resistance and extended-spectrum beta-lactamase-producing Enterobacteriaceae among hospitalized patients presumptive for bacterial infections at Debre Berhan Comprehensive Specialized Hospital, Ethiopia.MethodsA hospital-based cross-sectional study was conducted from January to May 2021. A total of 384 hospitalized patients presumptive for bacterial infections were included in the study. Urine, wound, blood, stool, and sputum samples were collected and cultured on MacConkey agar, Cysteine Lactose Electrolyte Deficient medium, and Blood agar. Identification was done using a panel of biochemical tests. The antimicrobial susceptibility test was done by disc diffusion. Screening of ESBL production was done by using cefotaxime and ceftazidime and confirmed by the combination disk method per clinical laboratory standard institute guidelines. Data analysis was performed by Statistical Package for Social Sciences software version 25, and a P-value ≤0.05 was considered as statistically significant.ResultsOut of 384 study participants, a total of 164 Enterobacteriaceae were isolated. The overall multi-drug resistance rate (MDR) was 92.1%. The overall prevalence of ESBL-PE was 104 (63.4%). E. coli 50 (30.5%) and K. pneumoniae 24 (14.6%) were the predominant ESBL producers. The highest ESBL producers E. coli (13.4%) and K. pneumoniae (6.1%) were isolated from urine sample. History of antibiotic use for the last three months (P-value=0.01), admission in neonatal intensive care unit (P-value=0.02), history of hospital stays (P-value=0.01), and chronic disease (P-value=0.04) showed statistically significant association with ESBL-PE infection.ConclusionThe prevalence of MDR-E and ESBL-PE was high. Therefore, strong infection prevention and control measures and careful selection of antibiotics are needed in the study area to block the transmission and infection in the healthcare setting.

Phenotypic Detection of ESBL, AmpC, MBL, and Their Co-occurrence among MDR Enterobacteriaceae Isolates.

Background Emergence of extended-spectrum beta-lactamases (ESBLs), AmpC β-lactamases, and metallo-β lactamases (MBL), and their co-existence among members of Enterobacteriaceae pose newer diagnostic and therapeutic challenges. The present study examines the ESBL, AmpC, and MBL production by various phenotypic methods and their co-occurrence among the multidrug-resistant (MDR) Enterobacteriaceae clinical isolates. Materials and Methods Four hundred non-repetitive Enterobacteriaceae clinical isolates were collected from the Central Referral Hospital, Sikkim. The isolates were used for identification and their antibiotic susceptibility tests were performed according to the Clinical and Laboratory Standard Institute (CLSI) guidelines. ESBL was detected by double-disc synergy test (DDST) and phenotypic confirmatory disc-diffusion test (PCDDT), AmpC detection by AmpC E-test, and boronic acid disc diffusion (BD) test. MBL was detected using the imipenem–imipenem/EDTA disc and carba-NP tests. Results Around 76% were considered MDR. ESBL was seen in 58% and 50.4% based on DDST and phenotypic confirmation disc-diffusion test (PCDDT), respectively. AmpC was detected in 11.8% and 13.1% using a commercial E-test and boronic acid test, respectively. MBL were identified in 12.8% and 14.8% based on MBL imipenem-EDTA and carba-NP tests, respectively. Co-occurrence of ESBL and AmpC, ESBL and MBL, AmpC and MBL was seen in 5.2%, 11.5%, 1.3%, respectively, whereas a combination of these three β-lactamases was observed in only 0.3% of 304 MDR isolates. Conclusion The findings highlight a high prevalence of β-lactamases and their co-production among the Enterobacteriaceae , mainly in Klebsiella pneumoniae and Escherichia coli isolates. The study further highlights the necessity to identify the MDR β-lactamases stains for effective therapy in severe as well as mild bacterial infections, thereby enabling to reduce the risk of MDR in hospital and community settings.

Clinical Feature, Therapy, Antimicrobial Resistance Gene Distribution, and Outcome of Nosocomial Meningitis Induced by Multidrug-Resistant Enterobacteriaceae-A Longitudinal Cohort Study From Two Neurosurgical Centers in Northern China.

ObjectivesThis is a comparative cohort study aiming to evaluate the mortality risk factors for patients with nosocomial meningitis (NM) induced by multidrug-resistant Enterobacteriaceae (MDRE) in China. The clinical features and therapies of patients and the resistance mechanisms of MDRE pathogens were also assessed.MethodsMDRE-NM patients from two neurosurgical centers in China from 2014 to 2019 were included in this study. Clinical features were extracted from the medical record databases of the two centers. The molecular mechanisms underlying the microbiological resistance mechanisms of each MDRE pathogen were determined, Kaplan–Meier survival analysis was conducted, and multivariable analyses were performed using a Cox proportional hazard model.ResultsNinety MDRE-NM patients were included in this study. Klebsiella pneumoniae accounted for the highest proportion of causative pathogens (46/90, 51.1%), and 40 causative pathogens (44.4%) were meropenem-resistant. blaKPC (27/40, 67.5%) was the predominant carbapenem resistance gene. Multivariate Cox analysis showed that external ventricular drainage (EVD) [hazard ratio (HR) = 2.524, 95% confidence interval (CI) = 1.101–5.787, p = 0.029] and a Glasgow Coma Scale (GCS) score ≤;8 (HR = 4.033, 95% CI = 1.526–10.645, p = 0.005) were mortality risk factors for patients with MDRE-NM. A total of 90.0%, 94.4%, and 97.8% of MDRE-NM patients received antibiotic prophylaxis (AP), antibiotic empirical therapy (AET), and antibiotic definitive therapy (ADT), respectively.ConclusionsNM caused by MDRE is an important sign of the failure of neurosurgery. MDRE possesses multiple drug resistance genotypes, and EVD and a GCS score ≤;8 are independent mortality risk factors for patients with MDRE-NM, which deserve the attention of microbiologists and neurosurgical clinicians.

Multidrug-Resistant Bacteria and Enterobacteriaceae Count in Abattoir Wastes and Its Receiving Waters in Limbe Municipality, Cameroon: Public Health Implications.

The release of untreated wastes from abattoirs into the environment and nearby water bodies poses a significant threat to public health. Such litters may contain pathogens, including antibiotic-resistant bacteria. This study investigated 80 samples collected from butchering tables, slaughter slabs, meat rinsing points, and abattoir wastes receiving water from two abattoirs (A and B). Total Enterobacteriaceae count (TEC) for each sample was determined, and Escherichia coli (E. coli), Salmonella spp., Shigella spp., and Staphylococcus aureus (S. aureus) were isolated and identified. Antimicrobial susceptibility testing was done on all bacterial isolates against nine locally used antibiotics. Overall, 118 bacterial isolates, comprising E. coli (42.5%), Salmonella spp. (27.5%), Shigella spp. (37.5%), and S. aureus (40.0%), were recovered. Of the 118 bacterial isolates, 104 (88.1%) were multidrug-resistant, including 58 (55.8%) from abattoir A and 46 (44.2%) from abattoir B; however, this difference was not statistically significant (p = 0.6837). Of the 32 S. aureus isolates, 29 (90.6%) were multidrug-resistant. All S. aureus were 100% sensitive to vancomycin, kanamycin, and amikacin. Similarly, 31 (91.2%) of the 34 E. coli isolates recovered in this study were multidrug-resistant. Salmonella spp. and Shigella spp. also showed high levels of multidrug resistance corresponding to 81.8% and 86.7%, respectively. All isolates of E. coli, Salmonella, and Shigella were 100% resistant to ampicillin and 100% sensitive to ciprofloxacin. Minimum and maximum mean values for TEC were 3.62-5.83 log CFU/mL for abattoir A and 4.08–5.56 log CFU/mL for abattoir B. The highest and lowest TEC counts were from slaughter slab and upstream water, respectively, in each abattoir. Our results indicate a predominance of multidrug-resistant bacteria in abattoir wastes and their receiving waters in the study sites. Hence, we recommend the treatment of abattoir wastes before disposal and improved hygiene and sanitation practices to enhance public health.

High prevalence of fecal carriage of Extended-spectrum beta-lactamase and carbapenemase-producing Enterobacteriaceae among food handlers at the University of Gondar, Northwest Ethiopia.

Fecal carriage of extended-spectrum beta-lactamase and Carbapenemase-producing Enterobacteriaceae is a potential risk for the transmission of infection with resistant strains. Understanding the burden of these resistant strains in asymptomatic people is essential to reduce the chain of infection transmission. However, data on the fecal carriage of Extended-spectrum Beta-lactamase and Carbapenemase-producing Enterobacteriaceae among food handlers were limited in developing countries especially in Ethiopia. The aim of the present study is, therefore, to assess fecal carriage rate, associated factors, and antimicrobial resistance patterns of Extended-spectrum Beta-lactamase and Carbapenemase-producing Enterobacteriaceae among food handlers at the University of Gondar Cafeterias, Northwest Ethiopia. An institution-based cross-sectional study was conducted from February to June 2021 at the University of Gondar cafeterias. A total of 290 stool samples were collected, transported using Cary Blair transport medium, and processed. All isolates were cultured and identified by using MacConkey agar, and routine biochemical tests. Antimicrobial susceptibility testing was done to each isolate following the Kirby Bauer disk diffusion method. If the zone of inhibition was ≤ 22 mm for ceftazidime, ≤25 mm for ceftriaxone, and ≤27 for cefotaxime they were considered as potential ESBL strain and selected for a further phenotypic confirmatory. Moreover, the double-disc diffusion test and the modified carbapenem inactivation method were used for confirmations of Extended-spectrum β-lactamase and Carbapenemase-producing Enterobacteriaceae respectively. If a ≥5mm difference in zone diameter for either antimicrobial agent in combination with clavulanic acid versus the zone diameter of the agent when tested alone (without B-lactamase inhibitor), was confirmed as ESBL-PE and if the zone of inhibition diameter between 6-15mm and 16- 18mm with a pinpoint colony, it was considered as carbapenem resistance Enterobacteriaceae. Data were entered using Epi-data version 4.6 and then exported to SPSS version 26 for analysis. Potential risk factors were assessed using multivariable logistic regression and a p-value less than 0.05 was considered statistically significant. Out of 290 stool samples, 63 (21.7%) and 7 (2.4%) were confirmed as Extended-spectrum β-lactamase and Carbapenemase-producing Enterobacteriaceae. The most predominant ESBL-PE was E. coli 43 (14.8%) followed by K. pneumoniae 17 (5.9%). Most of the Extended-spectrum β-lactamase and Carbapenemase-producing isolates were resistant to tetracycline, cefotaxime, ceftazidime, and ceftriaxone (100% each). In contrast, a low resistance level was recorded for Meropenem and cefoxitin. The overall Multi-drug resistant Enterobacteriaceae (MDR) was 147 (42.3%). Antibiotics usage in the last 3 months and drinking unpasteurized milk were associated with the carriage of the Extended-spectrum beta-lactamase-Producing Enterobacteriaceae. The high fecal carriage rate of Multi-drug resistance isolate, Extended-spectrum β-lactamase, and Carbapenemase-producing Enterobacteriaceae were recorded among food handlers. Therefore, this study gives signals in the spread of drug-resistant bacteria easily to the community. Hence, the need for adjusting and promotion of infection prevention measures to prevent the spread of drug-resistant bacteria should not be underestimated.

Molecular Detection and Antibiogram of ESBL-Producing and Carbapenem-Resistant Escherichia coli from Rabbit, Swine, and Poultry in Malaysia

The emergence of multidrug-resistance Enterobacteriaceae such as extended-spectrum β-lactamase (ESBL) producing Escherichia coli ( E. coli ) and carbapenem-resistant E. coli (CREC) has become an urgent veterinary and public health threat. These multidrug-resistant microorganisms are frequently associated with diseases that have high mortality with limited treatment options. This study aims to investigate the prevalence of ESBL producing E. coli and CREC from the rabbit, swine, and poultry and to determine the antibiogram profile of these E. coli isolates. In this study, 400 fecal swab samples were collected from rabbits, swine, and poultry from several selected animal farms in Malaysia. After incubation and isolation processes, suspected E. coli isolates were subjected to a PCR test to confirm the identity of the bacteria. The antibiogram of the E. coli isolates was determined via the Kirby Bauer disk diffusion method. A total of 212 (53%) E. coli isolates were isolated from rabbits (51 isolates), poultry (110 isolates), and swine (51 isolates). Screening of antimicrobial resistance genes revealed twelve ESBL producing E. coli (3%; 12/400). Two ESBL producing E. coli were also carrying carbapenemase gene (BlaNDM), indicating ESBL producing and carbapenem-resistant E. coli (ESBL-CREC) in poultry fecal swab samples. The bacteria isolates were found to show resistance against nine antibiotics, including ertapenem, ampicillin, and amoxicillin-clavulanate. A total of 3.3% (7/212) of the E. coli isolates were found to be multidrug-resistance. This study demonstrated the presence of ESBL-producing E. coli and ESBL-producing CREC from poultry fecal swabs in Malaysia.