Bismuth antimicrobial drugs serve as broad-spectrum metallo-ß-lactamase inhibitors.

Drug-resistant superbugs pose a huge threat to human health. Infections by Enterobacteriaceae producing metallo-ß-lactamases (MBLs), e.g., New Delhi metallo-ß-lactamase 1 (NDM-1) are very difficult to treat. Development of effective MBL inhibitors to revive the efficacy of existing antibiotics is highly desirable. However, such inhibitors are not clinically available till now. Here we show that an anti-Helicobacter pylori drug, colloidal bismuth subcitrate (CBS), and related Bi(III) compounds irreversibly inhibit different types of MBLs via the mechanism, with one Bi(III) displacing two Zn(II) ions as revealed by X-ray crystallography, leading to the release of Zn(II) cofactors. CBS restores meropenem (MER) efficacy against MBL-positive bacteria in vitro, and in mice infection model, importantly, also slows down the development of higher-level resistance in NDM-1-positive bacteria. This study demonstrates a high potential of Bi(III) compounds as the first broad-spectrum B1 MBL inhibitors to treat MBL-positive bacterial infection in conjunction with existing carbapenems.

A Novel Selective Medium for Isolation of Bacteroides fragilis from Clinical Specimens.

A novel Bacteroides fragilis selective (BFS) medium, consisting of a brain heart infusion agar base supplemented with yeast extract, cysteine hydrochloride, bile salts, vitamin K, hemin, glucose, esculin, ferric ammonium citrate, bromothymol blue, gentamicin, kanamycin, and novobiocin, was evaluated. When BFS agar was tested with a collection of 303 bacteria of different genera, it allowed the growth of B. fragilis as large yellow colonies, with blackening of the medium after 48 h of anaerobic incubation, while the growth of most other anaerobes, facultative anaerobes, and aerobes was inhibited. In a prospective comparison of BFS agar with a routinely used medium (neomycin blood agar) in 1,209 clinical specimens, 60 B. fragilis bacteria were detected on BFS agar while 46 were detected on the routine agar (McNemar's test, P = 0.008). In conclusion, this novel medium may be added to improve the recovery of B. fragilis in clinical specimens and to facilitate surveillance of antimicrobial-resistant strains.

Emergence of Macrolide-Resistant Mycoplasma pneumoniae in Hong Kong Is Linked to Increasing Macrolide Resistance in Multilocus Variable-Number Tandem-Repeat Analysis Type 4-5-7-2.

Macrolide-resistant Mycoplasma pneumoniae (MRMP) is rapidly emerging in Asia, but information on the temporal relationship between the increase in macrolide resistance and changes in strain types is scarce. Between 2011 and 2014, M. pneumoniae infection was diagnosed by PCR as part of routine care in a health care region in Hong Kong. Testing was initiated by clinicians, mainly in patients with suspected M. pneumoniae pneumonia. Specimens positive for M. pneumoniae were retrospectively investigated by macrolide resistance genotyping and a four-locus (Mpn13 to -16) multilocus variable-number tandem-repeat analysis (MLVA) scheme. The overall percentage of M. pneumoniae-positive specimens was 17.9%, with annual rates ranging from 9.8% to 27.2%. The prevalence of MRMP had rapidly increased from 13.6% in 2011 to 30.7% in 2012, 36.6% in 2013, and 47.1% in 2014 (P = 0.038). Two major MLVA types, 4-5-7-2 and 3-5-6-2, accounted for 75% to 85% of the infections each year. MLVA types 4-5-7-2 and 3-5-6-2 predominated among macrolide-resistant and macrolide-sensitive groups, respectively. The increase in MRMP was mainly caused by increasing macrolide resistance in the prevalent MLVA type 4-5-7-2, changing from 25.0% in 2011 to 59.1% in 2012, to 89.7% in 2013, and to 100% in 2014 (P < 0.001). In conclusion, increasing MRMP in Hong Kong was linked to a single MLVA type, which was both prevalent and increasingly resistant to macrolides.

Clinical outcome of extended-spectrum beta-lactamase-producing Escherichia coli bacteremia in an area with high endemicity.

OBJECTIVES: This study assessed the impact of discordant empirical antibiotic therapy on the outcome of bacteremia caused by extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli. METHODS: The clinical features and outcomes of a cohort of patients hospitalized with ESBL E. coli bacteremia between 2007 and 2008 were retrospectively reviewed. The effect of different antimicrobial regimens on patient outcomes was analyzed. RESULTS: ESBL E. coli accounted for 24.2% (207/857) of E. coli bacteremia cases. The urinary tract (43.6%) was the most common source of infection, followed by the hepatobiliary tract (23.0%). Discordant empirical antibiotic therapy was given to 52.0% patients. Admission to the intensive care unit was associated with the use of a carbapenem as empirical antibiotic therapy (p<0.001). Univariate analysis revealed no significant differences in 30-day mortality rates between patients receiving concordant and discordant empirical antibiotic therapy (23.5% vs. 19.8%, p=0.526), carbapenem and non-carbapenem empirical antibiotic therapy (29.8% vs. 19.1%, p=0.118), beta-lactam/beta-lactam inhibitor combinations (BLBLIs) and non-BLBLIs empirical antibiotic therapy (20.3% vs. 22.3%, p=0.734), cephalosporin and non-cephalosporin empirical antibiotic therapy (19.7% vs. 22.6%, p=0.639), and fluoroquinolone and non-fluoroquinolone empirical antibiotic therapy (8.3% vs. 22.4%, p=0.251). The findings were confirmed by multivariate analysis. CONCLUSIONS: Despite a high proportion of discordant empirical antibiotic therapy, ESBL production had little effect on 30-day mortality. Whether the observation can be applied to different ESBL types is unknown and warrants further study.

Epidemiology of Klebsiella oxytoca-associated diarrhea detected by Simmons citrate agar supplemented with inositol, tryptophan, and bile salts.

We studied the clinical and epidemiological characteristics of Klebsiella oxytoca-associated diarrhea in hospitalized patients in Hong Kong. Between 1 November 2009 and 30 April 2011, all inositol-fermenting colonies found on Simmons citrate agar supplemented with inositol, tryptophan, and bile salts (SCITB agar) used for the culturing of diarrheal stool samples were screened by a spot indole test for K. oxytoca. The overall sensitivity of SCITB agar plus the spot indole test (93.3%) for the detection of K. oxytoca in stool samples was superior to that of MacConkey agar (63.3%), while the specificities were 100% and 60.4%, respectively. The former achieved a 23-fold reduction in the workload and cost of subsequent standard biochemical identifications. Cytotoxin production and the clonality of K. oxytoca were determined by a cell culture cytotoxicity neutralization assay using HEp-2 cells and pulsed-field gel electrophoresis (PFGE), respectively. Of 5,581 stool samples from 3,537 patients, K. oxytoca was cultured from 117/5,581 (2.1%) stool samples from 104/3,537 (2.9%) patients. Seventy-six of 104 (73.1%) patients with K. oxytoca had no copathogens in their diarrheal stool samples. Twenty-four (31.6%) of 76 patients carried cytotoxin-producing strains, which were significantly associated with antibiotic therapy after hospital admission (50% versus 21.2%; P = 0.01). Health care-associated diarrhea was found in 44 (42%) of 104 patients with K. oxytoca, but there was no epidemiological linkage suggestive of a nosocomial outbreak, and PFGE showed a diverse pattern. None of the patients with cytotoxin-producing K. oxytoca developed antibiotic-associated hemorrhagic colitis, suggesting that K. oxytoca can cause a mild disease manifesting as uncomplicated antibiotic-associated diarrhea with winter seasonality.

Evolution of antibiotic resistance mechanisms and their relevance to dialysis-related infections.

As the survival of patients with end-stage renal failure has improved, their exposure to antibiotics has also increased. Infections, especially peritoneal dialysis-related peritonitis, are unavoidable because of lapses in technique and the slow worsening of systemic and peritoneal defense associated with aging and dialysis. The selective pressure inherent in the use of antibiotics shapes the pattern of antibiotic resistance in the bacteria causing peritonitis and extraperitoneal infections, and vice versa. Renal function-preserving and non-ototoxic regimens that incorporate double beta-lactams (first- and third-generation cephalosporins) for peritonitis have increased the selective pressure in favor of methicillin-resistant staphylococci (MRS) and extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae. Attempts to use the fluoroquinolones as alternatives to beta-lactams was met with rocketing quinolone resistance. The high incidence of MRS led many nephrologists to use empiric vancomycin-until the début of vancomycin-resistant enterococci. The recent emergence of heterogeneous and high-level vancomycin resistance in staphylococci (which are especially prevalent in patients on dialysis) calls for further prudence in the use of vancomycin. The coming challenges are ESBL-producing Enterobacteriaceae with carbapenemase, multi-resistant Pseudomonas, and highly virulent community-acquired methicillin-resistant Staphylococcus aureus with Panton-Valentine leukocidin. Antibiotic auditing programs and meticulous patient training by nurses are the only available defense at the moment. Novel approaches such as antibiotic-impregnated Tenckhoff catheters, biocompatible dialysis fluid, and peritoneal immuno-augmentation strategies are eagerly awaited.

Bacteremia caused by Escherichia coli producing extended-spectrum beta-lactamase: a case-control study of risk factors and outcomes.

A case-control study was conducted in order to identify the risk factors associated with bloodstream infection caused by Escherichia coli producing extended-spectrum beta-lactamase (ESBL) and to determine the outcomes of infected patients. Risk factors associated with ESBL production, according to univariate analysis, included a history of recent hospitalization [odds ratio (OR) 4.3, 95% confidence interval (CI) 2.1-8.9; p < 0.001], severe underlying diseases (OR 15, 95% CI 4.4-51.5; p < 0.001), prior exposure to urinary catheters (OR 8.3, 95% CI 3.2-21.7; p < 0.001) and nosocomial (OR 14.1, 95% CI 6.1-32.8; p < 0.001) or urinary (OR 3.6, 95% CI 1.7-7.4; p < 0.001) origin of the bacteria. Multivariate analysis revealed that severe underlying diseases (OR 31.2, 95% CI 6.7-144; p < 0.001) and nosocomial (OR 16.5, 95% CI 5.6-49; p < 0.001) and urinary origins (OR 7.8, 95% CI 2.6-23.8; p < 0.001) of the bacteria were independently associated with ESBL production in bacteremic E. coli. Crude mortality in case patients was more than twice as high as that in controls (p = 0.04). Production of ESBL increased the risk of inappropriate initial therapy (OR 95.6, 95% CI 27.4-334.2; p < 0.001). Treatment failed in 4/7 case patients treated with ceftazidime to which the isolate was susceptible in vitro. Our findings have implications for the choice of empirical therapy in nosocomial urinary tract infection.