Association Between Minimum Inhibitory Concentration, Beta-lactamase Genes and Mortality for Patients Treated With Piperacillin/Tazobactam or Meropenem From the MERINO Study.

INTRODUCTION: This study aims to assess the association of piperacillin/tazobactam and meropenem minimum inhibitory concentration (MIC) and beta-lactam resistance genes with mortality in the MERINO trial. METHODS: Blood culture isolates from enrolled patients were tested by broth microdilution and whole genome sequencing at a central laboratory. Multivariate logistic regression was performed to account for confounders. Absolute risk increase for 30-day mortality between treatment groups was calculated for the primary analysis (PA) and the microbiologic assessable (MA) populations. RESULTS: In total, 320 isolates from 379 enrolled patients were available with susceptibility to piperacillin/tazobactam 94% and meropenem 100%. The piperacillin/tazobactam nonsusceptible breakpoint (MIC >16 mg/L) best predicted 30-day mortality after accounting for confounders (odds ratio 14.9, 95% confidence interval [CI] 2.8-87.2). The absolute risk increase for 30-day mortality for patients treated with piperacillin/tazobactam compared with meropenem was 9% (95% CI 3%-15%) and 8% (95% CI 2%-15%) for the original PA population and the post hoc MA populations, which reduced to 5% (95% CI -1% to 10%) after excluding strains with piperacillin/tazobactam MIC values >16 mg/L. Isolates coharboring extended spectrum ß-lactamase (ESBL) and OXA-1 genes were associated with elevated piperacillin/tazobactam MICs and the highest risk increase in 30-day mortality of 14% (95% CI 2%-28%). CONCLUSIONS: After excluding nonsusceptible strains, the 30-day mortality difference from the MERINO trial was less pronounced for piperacillin/tazobactam. Poor reliability in susceptibility testing performance for piperacillin/tazobactam and the high prevalence of OXA coharboring ESBLs suggests that meropenem remains the preferred choice for definitive treatment of ceftriaxone nonsusceptible Escherichia coli and Klebsiella.

Levels of Mycoplasma genitalium Antimicrobial Resistance Differ by Both Region and Gender in the State of Queensland, Australia: Implications for Treatment Guidelines.

Mycoplasma genitalium is frequently associated with urogenital and rectal infections, with the number of cases of macrolide-resistant and quinolone-resistant M. genitalium infection continuing to increase. In this study, we examined the levels of resistance to these two common antibiotic treatments in geographically distinct locations in Queensland, Australia. Samples were screened for macrolide resistance-associated mutations using a commercially available kit (ResistancePlus MG; SpeeDx), and quinolone resistance-associated mutations were identified by PCR and DNA sequencing. Comparisons between antibiotic resistance mutations and location/gender were performed. The levels of M. genitalium macrolide resistance were high across both locations (62%). Quinolone resistance mutations were found in ∼10% of all samples, with a number of samples harboring mutations conferring resistance to both macrolides and quinolones. Quinolone resistance was higher in southeast Queensland than in north Queensland, and this was consistent in both males and females (P = 0.007). The M. genitalium isolates in rectal swab samples from males harbored high levels of macrolide (75.9%) and quinolone (19%) resistance, with 15.5% harboring resistance to both classes of antibiotics. Overall, the lowest observed level of resistance was to quinolones in females from north Queensland (1.6%). These data highlight the high levels of antibiotic resistance in M. genitalium isolates within Queensland and the challenges faced by sexually transmitted infection clinicians in managing these infections. The data do, however, show that the levels of antibiotic resistance may differ between populations within the same state, which has implications for clinical management and treatment guidelines. These findings also support the need for ongoing antibiotic resistance surveillance and tailored treatment.

Comparison of test specificities of commercial antigen-based assays and in-house PCR methods for detection of rotavirus in stool specimens.

Seven commercial rotavirus antigen assays were compared with in-house PCR methods for detecting rotavirus in stool specimens. The assay sensitivities were 80% to 100%, while the specificities were 54.3% for one commercial immunochromatographic (ICT) method and 99.4% to 100% for other assays. Thus, except for one commercial ICT, all the assays were generally reliable for rotavirus detection.

Molecular investigation of bacterial communities on the inner and outer surfaces of peripheral venous catheters.

Peripheral venous catheters (PVCs) are some of the most widely used medical devices in hospitals worldwide. PVC-related infections increase morbidity and treatment costs. The inner surfaces of PVCs are rarely examined for the population structure of bacteria, as it is generally believed that bacteria at this niche are similar to those on the external surface of PVCs. We primarily test this hypothesis and also study the effect of antibiotic treatment on bacterial communities from PVC surfaces. The inner and outer surfaces of PVCs from 15 patients were examined by 454 GS FLX Titanium 16S rRNA sequencing and the culture method. None of the PVCs were colonised according to the culture method and none of the patients had a bacteraemia. From a total of 127,536 high-quality sequence reads, 14 bacterial phyla and 268 diverse bacterial genera were detected. The number of operational taxonomic units for each sample was in the range of 86-157, even though 60 % of patients had received antibiotic treatment. Stenotrophomonas maltophilia was the predominant bacterial species in all the examined PVC samples. There were noticeable but not statistically significant differences between the inner and outer surfaces of PVCs in terms of the distribution of the taxonomic groups. In addition, the bacterial communities on PVCs from antibiotic-treated patients were significantly different from untreated patients. In conclusion, the surfaces of PVCs display complex bacterial communities. Although their significance has yet to be determined, these findings alter our perception of PVC-related infections.

Increases in Australian cutaneous abscess hospitalisations: 1999-2008.

Staphylococcus aureus is the most common cause of skin and soft tissue infections (SSTIs). Such infections have increased in several countries recently and at a time when community-associated methicillin-resistant S. aureus (CA-MRSA) strains have emerged globally. We examined changes in Australian hospitalisations for the treatment of cutaneous abscesses between 1999 and 2008, a period when increased numbers of CA-MRSA infections were being reported. National hospitalisation data for cutaneous abscess treatment (1999-2008) were examined. Hospitalisation numbers were collated and age-specific admission rates calculated and examined for changes over time. Yearly admissions for the treatment of cutaneous abscesses increased by 48%, from 8,849 (1999-2000) to 13,126 (2007-2008). The crude annual hospitalisation rate per 100,000 population rose from 46 to 62 respectively. However, increases in admission rates were limited to the 10 to 54 years age range. Incidence rate ratios (IRRs) for final versus baseline year admission rates for these age groups ranged from 1.36 (95% confidence interval [CI] 1.04-1.78) for those aged 10-14 years to 1.64 (95% CI 1.26-2.12) for those aged 45-49 years; p<0.05. Increases in hospitalisation for cutaneous abscess treatment have occurred in Australia during the last decade. Research into the underlying causes and prevention of these infections is a public health priority.

Influenza-associated bacterial pathogens in patients with 2009 influenza A (H1N1) infection: impact of community-associated methicillin-resistant Staphylococcus aureus in Queensland, Australia.

BACKGROUND: Secondary bacterial pneumonia due to community onset methicillin-resistant Staphylococcus aureus (MRSA) has become a highly publicised cause of influenza-associated death. There is a risk that case reports of fatal outcomes with post-influenza MRSA pneumonia may unduly influence antibiotic prescribing. AIMS: The aim of this study was to demonstrate the incidence of community-onset MRSA pneumonia in 2009 H1N1 influenza patients. METHODS: The microbiology records of patients positive for influenza A (H1N1) in 2009 were reviewed for positive blood or respiratory tract cultures and urinary pneumococcal antigen results within a Queensland database. Patients with such positive results within 48 h of hospital admission and a positive H1N1 influenza result in the prior 6 weeks were included. RESULTS: In 2009, 4491 laboratory-confirmed pandemic influenza A (H1N1) infections were detected. Fifty patients (1.1% of the H1N1 cohort) who were hospitalised with H1N1 and who had a bacterial respiratory tract pathogen were identified. Streptococcus pneumoniae (16 patients; 32%), Staphylococcus aureus (13 patients; 26%) and Haemophilus influenzae (9 patients; 18%) were the most commonly cultured organisms. Of the cohort of 4491 patients, MRSA was detected in only two patients, both of whom were admitted to intensive care units and survived after prolonged admissions. CONCLUSIONS: Influenza-associated community-onset MRSA pneumonia was infrequently identified in the 2009 H1N1 season in Queensland, despite community-onset MRSA skin and soft tissue infections being very common. Although post-influenza MRSA pneumonia is of great concern, its influence on empiric-prescribing guidelines should take into account its incidence relative to other secondary bacterial pathogens.

The role of surveillance cultures in the prediction of susceptibility patterns of Gram-negative bacilli in the intensive care unit.

Surveillance cultures may detect colonisation with drug-resistant Gram-negative bacteria and can be hypothesised to guide appropriate initial antibiotic treatment for intensive care unit (ICU) patients. We investigated the microbiological data of 228 episodes of nosocomial bloodstream infection (BSI) due to Gram-negative bacteria in an ICU in which piperacillin/tazobactam or meropenem was used empirically for serious infections, to evaluate the contribution of surveillance cultures to an appropriate choice of initial antibiotic therapy. Surveillance cultures were taken in advance of BSI in 218 (95.6%) of 228 episodes. Concordant organisms with identical identification and susceptibilities were found in prior surveillance cultures and subsequent blood cultures in 65 (29.8%) of 218 episodes. Surveillance cultures predicted resistance in 52.9% and 51.4% of BSIs caused by resistant pathogens to piperacillin/tazobactam and meropenem, respectively. The negative predictive value of surveillance cultures negative for a resistant organism also exceeded 90% for piperacillin/tazobactam and meropenem. Given that the overall resistant rates of BSI pathogens of our study were 11.3% to piperacillin/tazobactam and 16.4% to meropenem, surveillance cultures in our setting may provide important information on the probability of drug resistance of the causative pathogens and some utility in aiding empiric antibiotic therapy for ICU patients who subsequently develop BSI.

Epidemiology of non-multiresistant methicillin-resistant Staphylococcus aureus infection in Queensland, Australia: associations with indigenous populations and Panton-Valentine leukocidin.

The purpose of this study was to determine the extent of the spread of epidemic clones of non-multiresistant methicillin-resistant Staphylococcus aureus (nmMRSA) and the epidemiology of resultant infections throughout the state of Queensland. We collected a sample of clinical isolates of nmMRSA from laboratories serving public hospitals and clinics throughout the state. Three hundred isolates were typed and tested for the presence of Panton-Valentine leukocidin (PVL) genes and demographic and clinical data were collected from associated cases. Fifteen percent of S. aureus isolates were nmMRSA and 69% of these belonged to PVL-positive clones, predominantly ST93 and CC30. Low numbers of USA300- and USA400-like isolates were also present. Infections due to PVL-positive strains were much less frequently acquired in hospital (3.4%) than those due to PVL-negative nmMRSA (23.7%). Thirty-seven percent of cases were in indigenous people who make up only 3.6% of the general population. The proportion of cases with PVL-positive, but non-negative isolates decreased progressively with age, suggesting that immunity to PVL might be an important determinant of protection. nmMRSA strains are present throughout Queensland and cause infections in both community and healthcare settings.

First outbreak of PVL-positive nonmultiresistant MRSA in a neonatal ICU in Australia: comparison of MALDI-TOF and SNP-plus-binary gene typing.

The purpose of this brief report is to describe the first outbreak of a community-associated nonmultiresistant and PVL-positive MRSA strain (CC30) in a neonatal intensive care unit in Australia. The utility of matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF-MS) for microbial typing is compared with single nucleotide polymorphism (SNP) plus binary gene analysis. The composite correlation index analysis of the MALDI-TOF-MS data demonstrated the similar inter-strain relatedness found with the SNP-plus-binary gene typing used to confirm the outbreak. The evolving spread of MRSA emphasizes the importance of surveillance, infection control vigilance and the ongoing investigation of rapid typing methods for MRSA.

Geospatial epidemiology of Staphylococcus aureus in a tropical setting: an enabling digital surveillance platform.

Delivery of information to clinicians on evolving antimicrobial susceptibility needs to be accurate for the local needs, up-to-date and readily available at point of care. In northern Australia, bacterial infection rates are high but resistance to first- and second-line antibiotics is poorly described and currently-available datasets exclude primary healthcare data. We aimed to develop an online geospatial and interactive platform for aggregating, analysing and disseminating data on regional bacterial pathogen susceptibility. We report the epidemiology of Staphylococcus aureus as an example of the power of digital platforms to tackle the growing spread of antimicrobial resistance in a high-burden, geographically-sparse region and beyond. We developed an online geospatial platform called HOTspots that visualises antimicrobial susceptibility patterns and temporal trends. Data on clinically-important bacteria and their antibiotic susceptibility profiles were sought from retrospectively identified clinical specimens submitted to three participating pathology providers (96 unique tertiary and primary healthcare centres, n = 1,006,238 tests) between January 2008 and December 2017. Here we present data on S. aureus only. Data were available on specimen type, date and location of collection. Regions from the Australian Bureau of Statistics were used to provide spatial localisation. The online platform provides an engaging visual representation of spatial heterogeneity, demonstrating striking geographical variation in S. aureus susceptibility across northern Australia. Methicillin resistance rates vary from 46% in the west to 26% in the east. Plots generated by the platform show temporal trends in proportions of S. aureus resistant to methicillin and other antimicrobials across the three jurisdictions of northern Australia. A quarter of all, and up to 35% of methicillin-resistant S. aureus (MRSA) blood isolates in parts of the northern Australia were resistant to inducible-clindamycin. Clindamycin resistance rates in MRSA are worryingly high in regions of northern Australia and are a local impediment to empirical use of this agent for community MRSA. Visualising routinely collected laboratory data with digital platforms, allows clinicians, public health physicians and guideline developers to monitor and respond to antimicrobial resistance in a timely manner. Deployment of this platform into clinical practice supports national and global efforts to innovate traditional disease surveillance systems with the use of digital technology and to provide practical solutions to reducing the threat of antimicrobial resistance.

Prevalence of Staphylococcus aureus strains in an Australian cohort, 1989-2003: evidence for the low prevalence of the toxic shock toxin and Panton-Valentine leukocidin genes.

The purpose of this paper is to determine the prevalence of the toxic shock toxin gene (tst) and to enumerate the circulating strains of methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant S. aureus (MRSA) in Australian isolates collected over two decades. The aim was to subtype these strains using the binary genes pvl, cna, sdrE, pUB110 and pT181. Isolates were assayed using real-time polymerase chain reaction (PCR) for mecA, nuc, 16 S rRNA, eight single-nucleotide polymorphisms (SNPs) and for five binary genes. Two real-time PCR assays were developed for tst. The 90 MRSA isolates belonged to CC239 (39 in 1989, 38 in 1996 and ten in 2003), CC1 (two in 2003) and CC22 (one in 2003). The majority of the 210 MSSA isolates belonged to CC1 (26), CC5 (24) and CC78 (23). Only 18 isolates were tst-positive and only 15 were pvl-positive. Nine MSSA isolates belonged to five binary types of ST93, including two pvl-positive types. The proportion of tst-positive and pvl-positive isolates was low and no significant increase was demonstrated. Dominant MSSA clonal complexes were similar to those seen elsewhere, with the exception of CC78. CC239 MRSA (AUS-2/3) was the predominant MRSA but decreased significantly in prevalence, while CC22 (EMRSA-15) and CC1 (WA-1) emerged. Genetically diverse ST93 MSSA predated the emergence of ST93-MRSA (the Queensland clone).

Changing epidemiology of meticillin-resistant S. aureus in Queensland, Australia, 2000-2006: use of passive surveillance of susceptibility phenotypes.

The epidemiology of meticillin-resistant S. aureus (MRSA) infection has changed remarkably in recent years with the appearance of new MRSA strains causing infections in the community. These strains have now begun to cause healthcare-associated infections. The ability to track such changes is necessary to guide clinical and public health action. Here we report passive surveillance of all public laboratory susceptibility data in Queensland to track changes in MRSA phenotypes corresponding to the major epidemic strains from 2000 to 2006. The inpatient rate of MRSA isolation from pus, tissue and fluid (PTF) and blood culture (BC) specimens declined by 26% and 35%, respectively. The rate of isolation of the AUS-2/3-like phenotype (corresponding to ST239-MRSA-III) decreased from 651 to 242 isolates per million accrued patient days in inpatient PTF and BC, whereas that for non-multiresistant MRSA (nmMRSA, corresponding to community MRSA strains) increased from 71 to 315. The overall outpatient rate of MRSA isolation from PTF and BC increased by 224% and 31%, respectively. The rate of AUS-2/3-like isolates in outpatient PTF decreased from 131 to 60 per million outpatient occasions of service while the nmMRSA rate increased from 52 to 490. Surveillance of phenotypes derived from routine susceptibility data is a useful tool for tracking changes in the epidemiology of MRSA over large geographical regions.

Genomic insights into the emergence and spread of international clones of healthcare-, community- and livestock-associated meticillin-resistant Staphylococcus aureus: Blurring of the traditional definitions.

The evolution of meticillin-resistant Staphylococcus aureus (MRSA) from meticillin-susceptible S. aureus has been a result of the accumulation of genetic elements under selection pressure from antibiotics. The traditional classification of MRSA into healthcare-associated MRSA (HA-MRSA) and community-associated MRSA (CA-MRSA) is no longer relevant as there is significant overlap of identical clones between these groups, with an increasing recognition of human infection caused by livestock-associated MRSA (LA-MRSA). Genomic studies have enabled us to model the epidemiology of MRSA along these lines. In this review, we discuss the clinical relevance of genomic studies, particularly whole-genome sequencing, in the investigation of outbreaks. We also discuss the blurring of each of the three epidemiological groups (HA-MRSA, CA-MRSA and LA-MRSA), demonstrating the limited relevance of this classification.

Comparison of intravenous glucagon and dextrose in treatment of severe hypoglycemia in an accident and emergency department.

Hypoglycemia is a serious problem in insulin-treated diabetic patients. In this study the efficacy of intravenous glucagon (1 mg) was compared with that of intravenous dextrose (25 g) in the treatment of hypoglycemia in insulin-treated patients attending an accident and emergency department. In addition, the prevailing glycemic control of these patients was compared with patients routinely attending a diabetic outpatient clinic. Both intravenous glucagon and dextrose were effective in the treatment of hypoglycemic coma. There was a difference in the glycemic profile after intravenous glucagon compared with intravenous dextrose, and recovery of a normal level of consciousness after glucagon was slower than after dextrose (6.5 vs. 4.0 min, respectively; P less than .001), although the average duration of hypoglycemic coma was 1.4 h. The glucagon- and dextrose-treated groups had significantly lower HbA1 than comparable patients routinely attending the clinic (9.5 +/- 0.8 vs. 12.0 +/- 3.8%, respectively; P less than .001). In view of the ease of administration and the small risk of vascular and extravascular complications, intravenous glucagon appears to be a useful alternative to intravenous dextrose in the treatment of severe hypoglycemia.

ST2249-MRSA-III: a second major recombinant methicillin-resistant Staphylococcus aureus clone causing healthcare infection in the 1970s.

Typing of healthcare-associated methicillin-resistant Staphylococcus aureus (MRSA) from Australia in the 1970s revealed a novel clone, ST2249-MRSA-III (CC45), present from 1973 to 1979. This clone was present before the Australian epidemic caused by the recombinant clone, ST239-MRSA-III. This study aimed to characterize the genome of ST2249-MRSA-III to establish its relationship to other MRSA clones. DNA microarray analysis was conducted and a draft genome sequence of ST2249 was obtained. The recombinant structure of the ST2249 genome was revealed by comparisons to publicly available ST239 and ST45 genomes. Microarray analysis of genomic DNA of 13 ST2249 isolates showed gross similarities with the ST239 chromosome in a segment around the origin of replication and with ST45 for the remainder of the chromosome. Recombination breakpoints were precisely determined by the changing pattern of nucleotide polymorphisms in the genome sequence of ST2249 isolate SK1585 compared with ST239 and ST45. One breakpoint was identified to the right of oriC, between sites 1014 and 1065 of the gene D484_00045. Another was identified to the left of oriC, between sites 1185 and 1248 of D484_01632. These results indicate that ST2249 inherited approximately 35.3% of its chromosome from an ST239-like parent and 64.7% from an ST45-like parent. ST2249-MRSA-III resulted from a major recombination between parents that resemble ST239 and ST45. Although only limited Australian archival material is available, the oldest extant isolate of ST2249 predates the oldest Australian isolate of ST239 by 3 years. It is therefore plausible that these two recombinant clones were introduced into Australia separately.

Influence of antibody affinity on the performance of different antibody assays.

The effect of antibody affinity on the performance of 5 commonly used assays was studied. The assays used were measurement of antigen binding capacity in a Farr type assay, haemagglutination, solid-phase radioimmunoassay (SP-RIA), solid-phase ELISA and precipitation. The first 4 assays were all more sensitive for high affinity antibodies. Precipitation was not related to affinity, suggesting that factors secondary to antigen-antibody binding may be more important in determining the level of precipitate formation. The effect of epitope density of the antigen was also investigated in the SP-RIA and ELISA. Affinity dependence was more marked when antigen of low epitope density was used and this dependence could be reduced in the ELISA by choosing a low OD endpoint. Thus, the most reliable way to estimate antibody content by the ELISA may be to determine a low OD endpoint titre against antigen of high epitope density. When epitope density per molecule cannot be increased, an alternative approach to the problem is to increase epitope density by covalent coupling of antigen to the solid-phase rather than by adsorption.

Myoglobinuric acute renal failure associated with major urological surgery--an avoidable problem?

Recently, the problems associated with crush injury have recently been highlighted [1]. The association between major urological surgery and severe muscle damage is well recognised [2]. We report a case of myoglobinuric acute renal failure following radical cystectomy and suggest possible prevention.

The haemodynamic effects of intermittent haemofiltration in critically ill patients.

The haemodynamic effects of intermittent high volume venovenous haemofiltration were studied in 13 critically ill patients. The mean negative fluid balance during filtration was 1.21 and the mean duration of treatment 3 h 40 min. The cardiac index fell initially (4.5 +/- 0.2 to 3.8 +/- 0.21/min/m2; p less than 0.05) but then remained stable throughout treatment before returning to baseline at the end of haemofiltration. The mean arterial pressure was unchanged with an increase in the systemic vascular resistance (651 +/- 33 to 765 +/- 65 dyne.s/cm5; p less than 0.05) suggesting that vascular responsiveness is maintained during haemofiltration.

Adrenaline in treatment of septic shock: effects on haemodynamics and oxygen transport.

The effects of adrenaline on haemodynamics and oxygen transport were studied in 13 patients with septic shock persisting after optimal fluid loading. Adrenaline was administered by intravenous infusion at an increasing dose until no further benefit was seen. There were significant increases in mean arterial pressure, cardiac index, left ventricular stroke work index and oxygen delivery index. There was no significant change in oxygen consumption although the trend was towards an increase. There was a significant reduction in oxygen extraction ratio, but no change in shunt fraction. Adrenaline would appear to have beneficial haemodynamic effects in septic shock.