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Gastrointestinal carriage of Klebsiella pneumoniae is a predisposing factor for liver abscess in several Asian countries. To determine whether hypervirulent K. pneumoniae in the gut may be transmitted through food, we screened a range of raw and ready-to-eat retail food by culture and recovered K. pneumoniae in 21% (147 of 698) of samples tested. Based on PCR, no K. pneumoniae isolates carried the rmpA gene linked to community-acquired pyogenic liver abscess, providing no evidence of a link between food and liver disease. However, phenotypic resistance to multiple antibiotic classes was seen through disk diffusion tests, and carriage of genetic elements (wcaG and capsule types K1, K2, and K54) associated with increased virulence (8%, 11 of 147) was observed by PCR. Multidrug-resistant isolates were from raw vegetables, chicken or pork liver, and a ready-to-eat poultry dish; one multidrug-resistant K. pneumoniae isolate from raw bean sprouts was resistant to a third-generation cephalosporin (ceftriaxone). Although K. pneumoniae may be present in food without causing harm, we found isolates belonging to the K1 capsular serotype coexisting with the wcaG gene, one also conferring multidrug resistance. K. pneumoniae that carry antibiotic resistance genes, regardless of pathogenicity, may increase the available genetic pool of resistance along the food chain. Hygienic food handling practices are necessary to lower risks of acquiring K. pneumoniae and other opportunistic pathogens. Multidrug-resistant and potentially virulent K. pneumoniae was found in food. Food samples did not carry hypervirulent K. pneumoniae linked to liver abscess. Foodborne K. pneumoniae can be a potential reservoir of antibiotic resistance genes. Good food and personal hygiene are needed to help curb antimicrobial resistance. [ABSTRACT FROM AUTHOR]

Abstract: The purpose of this study was to characterise the metallo-β-lactamase VIM-6 and the genetic environment of the bla VIM-6 gene. The bla VIM-6 gene was cloned into an Escherichia coli expression system and the purified VIM-6 enzyme was obtained. VIM-6 has an isoelectric point of 4.9 and a molecular weight of 28.368Da. VIM-6 hydrolysed all tested penicillins, cephalosporins and carbapenems with the exception of aztreonam. In Pseudomonas putida, the bla VIM-6 gene is the first in a class 1 integron also containing bla OXA-10, aacA4, an open reading frame of unknown function, aadA and qacEΔ1. [Copyright &y& Elsevier]

• ERG11 mutations in Candida auris (C. auris) are clade-specific. • 42.9% of C. auris isolated in Singapore is multidrug-resistant. • The updated VITEKⓇ2 System has limited abilities for identifying C. auris. • Genomic data must be correlated with epidemiological information. Candida auris (C. auris) has globally emerged as a multidrug-resistant pathogen. While it is known that there are four geographic clades, little is known about its genomic epidemiology in the Southeast Asian region. Laboratory identification can be challenging but the VITEKⓇ2 system (version 8.01 software) has recently updated its database to include C. auris. This study aimed to investigate the genomic epidemiology of C. auris isolated in Singapore and the susceptibility profiles in relation to ERG11 and FKS1 mutations. Seven C. auris isolates from 2012–2018 were analysed using whole-genome sequencing, and antifungal susceptibility testing was performed. The performance of the updated VITEKⓇ2 system in identifying C. auris was also evaluated using these C. auris strains together with five closely related Candida species. Three clades were identified: South Asian (71.4%), South American (14.3%) and East Asian (14.3%). Local transmission was unlikely as there was no obviously identified cluster and most cases were likely to be imported at different time points following overseas hospitalisation exposure. Three isolates (42.9%) were multidrug-resistant. All South Asian strains were resistant to fluconazole and harboured ERG11 mutations, which were clade-specific. No FKS1 mutation was detected. The VITEKⓇ2 system was able to correctly identify most of the South Asian C. auris strains but misidentified the East Asian strain and gave a low discrimination result for the South American clade. This study showed that the introduction of C. auris into Singapore was possibly over multiple episodes and from different sources. The VITEKⓇ2 System version 8.01 software has limited abilities in identifying C.auris. [ABSTRACT FROM AUTHOR]

• Polymyxin resistance was most common in carbapenem resistant Enterobacter spp. • Prior polymyxin and carbapenem exposures were predictors of polymyxin resistance. • Many polymyxin-resistant cases were detected in patients not exposed to polymyxins. Increasing resistance to polymyxin, a last-line antibiotic, is a growing public health concern worldwide. The primary objective of this study was to identify predictors for the isolation of polymyxin-resistant (PR) carbapenem-resistant Enterobacteriaceae (CRE) among hospitalized patients. The secondary objective was to describe the clinical outcomes of patients with PR-CRE infections. A retrospective case–control study including patients admitted to Singapore General Hospital between June 2012 and June 2016 was conducted. Cases were defined as patients who had clinical cultures from which a PR-CRE was isolated. Controls were randomly selected from patients with polymyxin-susceptible (PS) CRE admitted during the same period, and frequency-matched to site of isolation. We included 37 PR cases and 111 PS controls. Polymyxin resistance was detected predominantly in Enterobacter spp. (54.1%) and Klebsiella pneumoniae (43.2%). Multilocus sequence typing showed little clonal relatedness among the isolates. mcr-1 was detected in two PR-CRE isolates. Multivariable analyses showed that PR-CRE isolation was associated with prior polymyxins (adjusted odds ratio (OR), 21.31; 95% confidence interval (CI), 3.04–150.96) and carbapenem exposures (OR 3.74; CI 1.13–12.44), when adjusted for time at risk and bacteria species. In PR-CRE patients with infections, the 30-day all-cause in-hospital mortality was 50.0% as compared to 38.1% in patients with PS-CRE (P = 0.346). Prior polymyxin and carbapenem exposures were independent risk factors for isolation of PR-CRE. Outcomes of PR-CRE and PS-CRE infections were similar in this study. [ABSTRACT FROM AUTHOR]

One of the most important resistance mechanisms in Gram-negative bacteria today is the production of enzymes causing resistance to cephalosporin and carbapenem antibiotics. The spread of extended-spectrum β-lactamases (ESBL)- and carbapenemase- producing Gram-negative bacteria is an emerging global public health problem. The aim of the present study was to (i) assess the prevalence of carbapenem-resistant bacteria (CRB) and ESBL-producing strains in sewage effluents from two major hospitals in Singapore, (ii) characterize the isolated strains and (iii) identify some of the ESBL and carbapenemase genes responsible for the resistance. CHROMagar ESBL and KPC plates were used to rapidly screen for ESBL-producing bacteria and those expressing reduced susceptibility to carbapenems, respectively. The abundance of ESBL-producers and CRB in hospital wastewater ranged between 10 3 and 10 6 CFU/mL. Out of the 66 isolates picked from ESBL and KPC plates, 95%, 82%, 82% and 76% were resistant to ceftriaxone, ceftazidime (3rd generation cephalosporin family), ertapenem and meropenem (carbapenem family), respectively. Among the resistant isolates, the most predominant taxa identified were Pseudomonas spp. (28.2%), Klebsiella spp. (28.2%), Enterobacter spp. (18.3%) and Citrobacter spp. (11.3%). PCR and sequencing analysis showed that the predominant β-lactamase genes were bla SHV (41.1%) followed by bla NDM-1 (35.6%), bla CTX (35.6%) and bla KPC (28.8%). The results of this study show a high prevalence of bacteria resistant to modern extended-spectrum cephalosporins and carbapenems and the presence of ESBL- and carbapenemase producers in hospital effluents. These findings support the need to improve management of hospital wastewater in order to minimize the spread of antimicrobial resistant microorganisms from this source. [ABSTRACT FROM AUTHOR]

Purpose: To describe an outbreak of Acanthamoeba keratitis (AK) cases among contact lens wearers. Design: Retrospective cohort study. Methods: Patients with AK were included. Relevant demographic and clinical data were obtained from case records, and patients were interviewed using a standardized questionnaire. Contact lens practices, including type of contact lens and solution used, were noted. In addition, clinical features at presentation, management, and clinical outcomes were recorded. Results: Forty-two patients (affecting 43 eyes) treated between 2000 and 2007 were included. Diagnosis was made by microbiologic culture in 35 cases and by microbiologic and histologic analysis in 2 cases, whereas the remainder were diagnosed based on clinical features and response to treatment. There was a gradual increase in cases since 2005, with a sharp increase in 2007, when 8 local patients were treated. Of 30 patients where contact lens solution data were available, 18 reported using a Complete brand Multipurpose solution (Advanced Medical Optics, Santa Ana, California, USA) before the infection. Among resident cases treated since February 2006, 7 (63%) of 11 patients used a Complete brand solution. Suboptimal hygiene practices were found in all patients interviewed. Fifteen patients required corneal grafting, with 11 undergoing therapeutic deep lamellar keratoplasty (DLK), 2 undergoing optical penetrating keratoplasty (PK), 1 undergoing optical DLK, and 1 undergoing therapeutic PK. The remainder were treated successfully medically with combination antiamebic therapy. The average duration of therapy was 116.2 days (range, 15 to 283 days). Of patients with radial keratoneuritis with or without epithelial disease, 83.3% achieved final vision of 20/40 or better, whereas this was achieved in 41.7% of those with ring infiltrate. Twenty-five percent of patients with ring infiltrate had final visual acuity of counting fingers or worse, whereas no patient with keratoneuritis and epithelial disease had final vision worse than counting fingers. Conclusions: There was an increase in the number of contact lens users with AK seen in the major eye departments of Singapore. Most of our patients also reported using a Complete brand Multipurpose solution before infection, and this parallels a similar outbreak in the United States. Increasing severity of infection was associated with worse visual outcome. [Copyright &y& Elsevier]