Invasive group A streptococcal disease surveillance in Canada, 2021-2022.

Background: Invasive group A streptococcal (iGAS, Streptococcus pyogenes) disease has been a nationally notifiable disease in Canada since 2000. This report summarizes the demographics, emm types, and antimicrobial resistance of iGAS isolates collected in Canada in 2021 and 2022. Methods: The Public Health Agency of Canada's National Microbiology Laboratory collaborates with provincial and territorial public health laboratories to conduct national surveillance of invasive S. pyogenes. Emm typing was performed using the Centers for Disease Control and Prevention emm sequencing protocol or extracted from whole-genome sequencing data. Antimicrobial susceptibilities were determined using Kirby-Bauer disk diffusion according to Clinical and Laboratory Standards Institute guidelines or predicted from whole-genome sequencing data based on the presence of resistance determinants. Results: Overall, the incidence of iGAS disease in Canada was 5.56 cases per 100,000 population in 2021, decreasing from the peak of 8.6 cases per 100,000 population in 2018. A total of 2,630 iGAS isolates were collected during 2022, representing an increase from 2021 (n=2,179). In particular, there was a large increase in isolates collected from October to December 2022. The most predominant emm type overall in 2021 and 2022 was emm49, at 21.5% (n=468) and 16.9% (n=444), respectively, representing a significant increase in prevalence since 2018 (p<0.0001). The former most prevalent type, emm1, increased from 0.5% (n=10) in 2021 to 4.8% (n=125) in 2022; similarly, emm12 increased from 1.0% (n=22) in 2021 to 5.8% (n=151) in 2022. These two types together accounted for almost 25% of isolates collected in late 2022 (October to December). Antimicrobial resistance rates in 2021 and 2022 included: 14.9%/14.1% erythromycin resistance, 4.8%/3.0% clindamycin resistance, and <1% chloramphenicol resistance. Conclusion: The increase of iGAS isolates collected in Canada is an important public health concern. Continued surveillance of iGAS is critical to monitor expanding emm types and antimicrobial resistance patterns.

The microbiological outcomes of culture-negative blood specimens using 16S rRNA broad-range PCR sequencing: a retrospective study in a Canadian province from 2018 to 2022.

Broad-range 16S rRNA PCR and sequencing of 1,183 blood specimens from 853 unique patients yielded an interpretable sequence and bacterial identification in 29%, 16S rRNA amplification with uninterpretable sequences in 53%, and no amplification in 18%. This study highlights the potential utility of this technique in identifying fastidious gram-negative and anaerobic bacteria but the frequent recovery of environmental and contaminant organisms argues for its judicious use. IMPORTANCE: The existing literature focuses on its performance compared to blood cultures in patients with sepsis, leaving a gap in the literature regarding other blood specimens in suspected infectious syndrome across the severity spectrum. We aimed to characterize its microbiological outcomes and provide insight into its potential clinical utility.

Oral Sex following Abortion: Case Report of a Sexually Transmitted Infection of Group A Streptococcus Causing Peritonitis.

Group A Streptococcus (GAS) is a rare cause of peritonitis with only a few reports of disease associated with surgical abortion, vaginal delivery, or intrauterine devices, most of which are speculated to be in association with the female genital tract. Only a single case of GAS infection transmission through contemporary oral sex has been previously reported. We report a strange case of GAS peritonitis occurring after abortion and oral sex.

Invasive group A streptococcal disease surveillance in Canada, 2020.

Background: Invasive group A streptococcal (iGAS) disease (caused by Streptococcus pyogenes) has been a nationally notifiable disease in Canada since 2000. This report summarizes the demographics, emm types and antimicrobial resistance of iGAS infections in Canada in 2020. Methods: The Public Health Agency of Canada's National Microbiology Laboratory (Winnipeg, Manitoba) collaborates with provincial and territorial public health laboratories to conduct national surveillance of invasive S. pyogenes. Emm typing was performed on all isolates using the Centers for Disease Control and Prevention emm sequencing protocol. Antimicrobial susceptibilities were determined using Kirby-Bauer disk diffusion according to Clinical and Laboratory Standards Institute guidelines. Population-based iGAS disease incidence rates up to 2019 were obtained through the Canadian Notifiable Disease Surveillance System. Results: Overall, the incidence of iGAS disease in Canada has increased from 4.0 to 8.1 cases per 100,000 population from 2009 to 2019. The 2019 incidence represents a slight decrease from the 2018 rate of 8.6 cases per 100,000 population. A total of 2,867 invasive S. pyogenes isolates that were collected during 2020 are included in this report, representing a decrease from 2019 (n=3,194). The most common emm types in 2020 were emm49 (16.8%, n=483) and emm76 (15.0%, n=429), both increasing significantly in prevalence since 2016 (p<0.001). The former most prevalent type, emm1, decreased to 7.6% (n=217) in 2020 from 15.4% (n=325) in 2016. Antimicrobial resistance rates in 2020 included 11.5% resistance to erythromycin, 3.2% resistance to clindamycin and 1.6% nonsusceptibility to chloramphenicol. Conclusion: Though the number of collected invasive S. pyogenes isolates decreased slightly in 2020 in comparison to previous years, iGAS disease remains an important public health concern. The emm distribution in Canada has been subtly shifting over the past five years, away from common and well-known emm1 and towards emm49 and emm76. It is important to continue surveillance of S. pyogenes in Canada to monitor expanding replacement emm types, as well as outbreak clones and antimicrobial resistance.

Validation of a Gradient Diffusion Method (Etest) for Testing of Antimicrobial Susceptibility of Aerococcus urinae to Fluoroquinolones.

Aerococcus urinae is a urinary pathogen with well-described resistance to fluoroquinolones. This study aimed to validate the gradient diffusion (GD) method (Etest) on cation-adjusted Mueller-Hinton agar with 5% sheep blood for testing the susceptibilities of Aerococcus urinae to the antimicrobial agents ciprofloxacin and levofloxacin and to compare the Etest to the broth microdilution (BMD) method from CLSI document M45-A3. Agar dilution (AD), as recommended by EUCAST, was used as an alternative reference method to arbitrate discrepancies or address technical issues. Aerococcus urinae isolates from urinary specimens were prospectively collected between June 2016 and December 2017 from six hospitals in Quebec, Canada, and identifications were confirmed using Vitek MS with the IVD 3.0 database. Of the 207 isolates tested using BMD, 37 (17.9%) showed trailing and 19 (9.2%) showed insufficient growth; these were tested using AD. Also, 38 isolates (18.4%) for ciprofloxacin and 13 isolates (6.3%) for levofloxacin showed a lack of essential or categorical agreement between the Etest and BMD and were also tested by AD. By use of a combined reference method (BMD or AD), the susceptibility rates of Aerococcus urinae were 82.6% and 81.6% for ciprofloxacin and levofloxacin, respectively. Categorical agreement between GD and the combined reference methods was 95.2% for ciprofloxacin and 97.1% for levofloxacin, with no very major error identified. Major and minor error rates were 0.6% and 4.3% for ciprofloxacin and 1.2% and 1.9% for levofloxacin. Overall, antimicrobial susceptibility testing (AST) using the Etest on sheep blood agar showed good agreement with the reference methods and can be considered by clinical laboratories wishing to perform AST on Aerococcus urinae isolates.

Enemella gen. nov., Enemella evansiae sp. nov., Enemella dayhoffiae sp. nov. and Parenemella sanctibonifatiensis gen. nov., sp. nov., novel taxa assignable to the family Propionibacteriaceae and derived from human clinical samples.

Nine Gram-stain-positive cocci, coccobacilli or short, rod-shaped strains recovered from clinical sources from patients located in two Canadian provinces and one environmental source were extensively studied. Clinical sources included blood cultures, cerebral spinal fluid, lymph node, lung biopsy and peritoneal fluid. Through 16S rRNA gene and whole genome sequencing analyses, the strains were found to cluster into three groups, closest to but distinguished from other genera in the family Propionibacteriaceae. The genomes from these bacteria had high G+C content, ranging from 67.8-69.56 mol%, and genome sizes of 3.02-4.52 Mb. Biochemical and chemotaxonomic properties including branched-chain cellular fatty acids, l-lysine diaminopimelic acid (ll-DAP) and cell-wall type A3γ (ll-DAP-gly) containing ll-DAP, alanine, glycine and glutamic acid were found and so the strains were therefore deemed to be consistent with other new genera in this family. Based on this investigation, we propose Enemella gen. nov., Enemella evansiae sp. nov., Enemella dayhoffiae sp. nov. and Parenemella sanctibonifatiensis gen. nov., sp. nov. for these taxa. Misidentified taxon 'Ponticoccus gilvus' was found to be assignable to Enemella evansiae based on this study.

Bacillus cereus infection in neonates and the absence of evidence for the role of banked human milk: Case reports and literature review.

BACKGROUND: Banked human milk (BHM) has inherent infectious risks, even when pasteurized. Because of the ubiquity of Bacillus cereus in the environment and its ability to resist the Holder pasteurization process, there is a concern that BHM might lead to severe B. cereus infections. OBJECTIVE: We reviewed observed and published cases to determine the potential causal role of BHM as the source of these infections. METHODS: Two infants in the province of Québec (Canada) developed a B. cereus neonatal infection, and both had received BHM. We conducted bacteriological studies to compare clinical isolates and those found in these cases. RESULTS: After extended culture of BHM retention lots, B. cereus was found to have been involved in batches related to the first case. However, molecular typing showed that the strain was different from the clinical isolate, therefore excluding BHM as the source of contamination. In the second case, a Brevibacillus spp was isolated, a species distinct from the clinical isolate. CONCLUSION: Based on these cases and others reported in the literature, a causal link between B. cereus contaminated BHM and preterm neonatal infection has never been documented. Therefore, the risk that BHM can cause this infection remains theoretical. Given the widespread presence of B. cereus in the hospital environment and its capacity to resist standard cleaning procedures, it seems likely that airborne or direct or indirect contact are the main sources of most, if not all, cases of severe B. cereus neonatal infections, even in babies exposed to BHM.

Criibacterium bergeronii gen. nov., sp. nov., a new member of the family Peptostreptococcaceae, isolated from human clinical samples.

A rod-shaped, motile anaerobic bacterium, designated CCRI-22567T, was isolated from a vaginal sample of a woman diagnosed with bacterial vaginosis and subjected to a polyphasic taxonomic study. The novel strain was capable of growth at 30-42 °C (optimum, 42 °C), at pH 5.5-8.5 (optimum, pH 7.0-7.5) and in the presence of 0-1.5 % (w/v) NaCl (optimally at 0.5 % NaCl). The phylogenetic trees based on 16S rRNA gene sequences showed that strain CCRI-22567T forms a distinct evolutionary lineage independent of other taxa in the family Peptostreptococcaceae. Strain CCRI-22567T exhibited 90.1 % 16S rRNA gene sequence similarity to Peptoanaerobacter stomatis ACC19aT and 89.7 % to Eubacterium yurii subsp. schtitka ATCC 43716. The three closest organisms with an available whole genome were compared to strain CCRI-22567T for genomic relatedness assessment. The genomic average nucleotide identities (OrthoANIu) obtained with Peptoanaerobacter stomatis ACC19aT, Eubacterium yurii subsp. margaretiae ATCC 43715 and Filifactor alocis ATCC 35896T were 71.8, 70.3 and 69.6 %, respectively. Strain CCRI-22567T contained C18 : 1 ω9c and C18 : 1 ω9c DMA as the major fatty acids. The DNA G+C content of strain CCRI-22567T based on its genome sequence was 33.8 mol%. On the basis of the phylogenetic, chemotaxonomic and other phenotypic properties, strain CCRI-22567T is considered to represent a new genus and species within the family Peptostreptococcaceae, for which the name Criibacterium bergeronii gen. nov., sp. nov., is proposed. The type strain of Criibacterium bergeronii is CCRI-22567T (=LMG 31278T=DSM 107614T=CCUG 72594T).

Group A Streptococcus Outbreak in a Canadian Armed Forces Training Facility.

Between December 2016 and April 2017, two cases of invasive Group A Streptococcus (GAS) infections were reported at a Canadian military training facility. An outbreak was declared and a field investigation was launched to characterize the outbreak and identify associated risk factors to limit transmission. Throat culture data from military personnel at the garrison were analyzed. Investigators tracked invasive GAS cases and non-invasive hospitalized GAS cases, and conducted site visits and case interviews. Sensitivity and specificity for a rapid antigen detection test were evaluated. Molecular typing and phylogenomic relationships of outbreak isolates were analyzed using whole-genome sequencing. During this outbreak, four invasive cases were reported and six non-invasive cases were hospitalized. In a sample of 705 throat cultures examined, 35.2% were GAS-positive. Among 65 platoon contacts of one invasive case, 30.2% were GAS-positive. Reluctance to seek medical care, challenges in following cough etiquette, and low compliance with antibiotics were identified among recruits. The rapid antigen detection test had low sensitivity (31.6%) during the outbreak. The outbreak sequence type was emm6.4 and outbreak isolates were highly related phylogenetically, differing by 0-4 single nucleotide variants. This is the first report of a GAS outbreak among Canadian military trainees. Increased surveillance of GAS infections, increased control measures and outbreak-specific clinical guidelines were implemented in-garrison. No further invasive GAS cases were identified. A GAS surveillance system was implemented and efforts to improve antibiotic compliance and medical consultation were recommended.

Canada-Wide Epidemic of emm74 Group A Streptococcus Invasive Disease.

BACKGROUND: The number of invasive group A Streptococcus (iGAS) infections due to hitherto extremely rare type emm74 strains has increased in several Canadian provinces since late 2015. We hypothesized that the cases recorded in the different provinces are linked and caused by strains of an emm74 clone that recently emerged and expanded explosively. METHODS: We analyzed both active and passive surveillance data for iGAS infections and used whole-genome sequencing to investigate the phylogenetic relationships of the emm74 strains responsible for these invasive infections country-wide. RESULTS: Genome analysis showed that highly clonal emm74 strains, genetically different from emm74 organisms previously circulating in Canada, were responsible for a country-wide epidemic of >160 invasive disease cases. The emerging clone belonged to multilocus sequence typing ST120. The analysis also revealed dissemination patterns of emm74 subclonal lineages across Canadian provinces. Clinical data analysis indicated that the emm74 epidemic disproportionally affected middle-aged or older male individuals. Homelessness, alcohol abuse, and intravenous drug usage were significantly associated with invasive emm74 infections. CONCLUSIONS: In a period of 20 months, an emm74 GAS clone emerged and rapidly spread across several Canadian provinces located more than 4500 km apart, causing invasive infections primarily among disadvantaged persons.

Comparison of sequential multiplex PCR, sequetyping and whole genome sequencing for serotyping of Streptococcus pneumoniae.

Streptococcus pneumoniae is one of the major causes of pneumonia, meningitis and other pneumococcal infections in young children and elders. Determination of circulating S. pneumoniae serotypes is an essential service by public health laboratories for the monitoring of putative serotype replacement following the introduction of pneumococcal conjugate vaccines (PCVs) and of the efficacy of the immunization program. The Quellung method remains the gold standard for typing S. pneumoniae. Although this method is very effective, it is also costly, time consuming and not totally reliable due to its subjective nature. The objectives of this study were to test and evaluate the efficiency of 3 different molecular methods compared to the Quellung method. Sequential multiplex PCR, sequetyping and whole genome sequencing (WGS) were chosen and tested using a set of diverse S. pneumoniae. One-hundred and eighteen isolates covering 83 serotypes were subjected to multiplex PCR and sequetyping while 88 isolates covering 53 serotypes were subjected to WGS. Sequential multiplex PCR allowed the identification of a significant proportion (49%) of serotypes at the serogroup or subset level but only 27% were identified at the serotype level. Using WGS, 55% to 60% of isolates were identified at the serotype level depending on the analysis strategy used. Finally, sequetyping demonstrated the lowest performance, with 17% of misidentified serotypes. The use of Jin cpsB database instead of the GenBank database slightly improved results but did not significantly impact the efficiency of sequetyping. Although none of these molecular methods may currently replace the Quellung method, WGS remains the most promising molecular pneumococcal serotyping method.

A case report of a deep surgical site infection with Terrisporobacter glycolicus/T. Mayombei and review of the literature.

BACKGROUND: There are increasing data regarding Terrisporobacter glycolicus as an emerging anaerobic pathogen. However, the few published cases to date usually report it as part of a polymicrobial infection. Here, we describe the first reported monomicrobial surgical site infection with this bacterium. Identification methods, taxonomy, and clinical management of this rarely identified pathogen are also discussed. CASE PRESENTATION: A previously healthy 66-year-old sustained an open olecranon fracture of his left arm after trauma. He subsequently underwent open reduction and internal fixation (ORIF), with insertion of an olecranon locking plate and two locking screws. Ten days after surgery, the patient developed increasing pain at the surgical site and noted green discharge from the wound. Culture of the wound discharge yielded grew a pure Gram-positive anaerobe identified by the RapidANA® microbial identification system as C. difficile (profile 000010, 99.1 % probability). Reference laboratory testing identified the isolate as T. glycolicus/mayombei (previously designated as Clostridium glycolicum/mayombei) by 16S rRNA gene sequencing and as Clostridium glycolicum by MALDI-TOF mass spectrometry. The patient received an 8-week course of moxifloxacin and metronidazole with an excellent clinical response at 12 months' follow-up. CONCLUSIONS: We describe the case of a deep surgical site infection with T. glycolicus/mayombei (formerly known as Clostridium glycolicum and Clostridium mayombei, respectively), which extends our knowledge of the clinical spectrum of this pathogen. The isolate was misidentified by phenotypic identification methods.

First Report of Clostridium lavalense Isolated in Human Blood Cultures.

An 88-year-old man was admitted to the hospital with worsening malaise, fever, and weakness. Anaerobic blood culture bottles revealed the presence of an anaerobic, Gram-positive sporulated bacillus. Empirical antibiotherapy with intravenous piperacillin-tazobactam was initiated. The patient defervesced after four days and was switched to oral amoxicillin on his 6th day of antibiotic therapy and later discharged from the hospital. Four months later, he had recovered. The bacterium was initially identified as Clostridium butyricum using anaerobic manual identification panel. 16S rRNA gene sequence and phylogenetic analysis showed the bacterium to be Clostridium lavalense, a recently described species with no previously published case of isolation in human diagnostic samples so far. This is the first report of Clostridium lavalense isolation from human blood cultures. Further studies are needed in order to elucidate the role of Clostridium lavalense in human disease and its virulence factors.

Escherichia coli antimicrobial susceptibility profile and cumulative antibiogram to guide empirical treatment of uncomplicated urinary tract infections in women in the province of Québec, 2010-15.

OBJECTIVES: Empirical treatment of uncomplicated urinary tract infections (UTIs) in women should be based on local susceptibility data. We aimed to generate regional and provincial cumulative antibiograms combining data from different laboratory information systems and determine the impact of basic patient characteristics on susceptibility results. METHODS: All positive urine samples for Escherichia coli obtained from women aged 18-65 years old in outpatient settings between 1 April 2010 and 31 March 2015 from four hospitals in Quebec, Canada, were included. The cumulative antibiogram for ciprofloxacin, nitrofurantoin and trimethoprim/sulfamethoxazole was calculated. A clinically significant difference in susceptibility profile was defined as factor(s) that lowered the susceptibility proportion below 80%. RESULTS: A total of 36 293 positive urine cultures were analysed. In the last year of the study, the proportion of susceptibility for ciprofloxacin, nitrofurantoin and trimethoprim/sulfamethoxazole was 90.3%, 95.4% and 81.9%, respectively. The susceptibility proportion was <80% for trimethoprim/sulfamethoxazole in the Montreal region (73.4%; 95% CI 71.1%-75.9%), whereas it remained >80% for the other regions. A significant decrease in susceptibility with time was identified for ciprofloxacin (92.1%-90.3%, P < 0.001) and nitrofurantoin (97.1%-95.4%, P < 0.001). Increasing age, recent hospitalization and site of collection were associated with an increase in resistance for certain antibiotics. CONCLUSIONS: Overall, all first-line antimicrobials remain acceptable choices for empirical treatment of uncomplicated UTIs in women in Quebec. The regional variability in susceptibility data within a single province emphasizes the importance of local susceptibility data to inform the development of empirical treatment guidelines for UTIs.

The initial state of the human gut microbiome determines its reshaping by antibiotics.

Microbiome studies have demonstrated the high inter-individual diversity of the gut microbiota. However, how the initial composition of the microbiome affects the impact of antibiotics on microbial communities is relatively unexplored. To specifically address this question, we administered a second-generation cephalosporin, cefprozil, to healthy volunteers. Stool samples gathered before antibiotic exposure, at the end of the treatment and 3 months later were analysed using shotgun metagenomic sequencing. On average, 15 billion nucleotides were sequenced for each sample. We show that standard antibiotic treatment can alter the gut microbiome in a specific, reproducible and predictable manner. The most consistent effect of the antibiotic was the increase of Lachnoclostridium bolteae in 16 out of the 18 cefprozil-exposed participants. Strikingly, we identified a subgroup of participants who were enriched in the opportunistic pathogen Enterobacter cloacae after exposure to the antibiotic, an effect linked to lower initial microbiome diversity and to a Bacteroides enterotype. Although the resistance gene content of participants' microbiomes was altered by the antibiotic, the impact of cefprozil remained specific to individual participants. Resistance genes that were not detectable prior to treatment were observed after a 7-day course of antibiotic administration. Specifically, point mutations in beta-lactamase blaCfxA-6 were enriched after antibiotic treatment in several participants. This suggests that monitoring the initial composition of the microbiome before treatment could assist in the prevention of some of the adverse effects associated with antibiotics or other treatments.

Enteroccocus pallens as a potential novel human pathogen: three cases of spontaneous bacterial peritonitis.

INTRODUCTION: Enterococcus pallens is one of the four yellow-pigmented members of the genus Enterococcus. To date, a single report of E. pallens isolated from a human sample has been published. CASE PRESENTATION: We report three cases of E. pallens spontaneous bacterial peritonitis in patients with liver cirrhosis that all occurred in Quebec, Canada. Ascitic fluid analysis revealed the presence of E. pallens in culture. Identification was made by classical biochemical testing and MALDI-TOF MS, as well as 16S rRNA and elongation factor (tuf) gene sequencing. Two of the three patients recovered after antimicrobial treatment. CONCLUSION: This report identifies E. pallens as a novel human pathogen that appears to possess particular but as-yet unidentified virulence factors that favour the development of peritoneal fluid infections, as previously reported for other Enterococcus species. Clinical microbiologist should be aware of this micro-organism which can be identified by phenotypic and molecular methods.

A Side by Side Comparison of Bruker Biotyper and VITEK MS: Utility of MALDI-TOF MS Technology for Microorganism Identification in a Public Health Reference Laboratory.

Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has emerged as a rapid, highly accurate, and cost-effective method for routine identification of a wide range of microorganisms. We carried out a side by side comparative evaluation of the performance of Bruker Biotyper versus VITEK MS for identification of a large and diverse collection of microorganisms. Most difficult and/or unusual microorganisms, as well as commonly encountered microorganisms were selected, including Gram-positive and negative bacteria, mycobacteria, actinomycetes, yeasts and filamentous fungi. Six hundred forty two strains representing 159 genera and 441 species from clinical specimens previously identified at the Laboratoire de santé publique du Québec (LSPQ) by reference methods were retrospectively chosen for the study. They included 254 Gram-positive bacteria, 167 Gram-negative bacteria, 109 mycobacteria and aerobic actinomycetes and 112 yeasts and moulds. MALDI-TOF MS analyses were performed on both systems according to the manufacturer's instructions. Of the 642 strains tested, the name of the genus and / or species of 572 strains were referenced in the Bruker database while 406 were present in the VITEK MS IVD database. The Biotyper correctly identified 494 (86.4%) of the strains, while the VITEK MS correctly identified 362 (92.3%) of the strains (excluding 14 mycobacteria that were not tested). Of the 70 strains not present in the Bruker database at the species level, the Biotyper correctly identified 10 (14.3%) to the genus level and 2 (2.9%) to the complex/group level. For 52 (74.2%) strains, we obtained no identification, and an incorrect identification was given for 6 (8.6%) strains. Of the 178 strains not present in the VITEK MS IVD database at the species level (excluding 71 untested mycobacteria and actinomycetes), the VITEK MS correctly identified 12 (6.8%) of the strains each to the genus and to the complex/group level. For 97 (54.5%) strains, no identification was given and for 69 (38.7%) strains, an incorrect identification was obtained. Our study demonstrates that both systems gave a high level (above 85%) of correct identification for a wide range of microorganisms. However, VITEK MS gave more misidentification when the microorganism analysed was not present in the database, compared to Bruker Biotyper. This should be taken into account when this technology is used alone for microorganism identification in a public health laboratory, where isolates received are often difficult to identify and/or unusual microorganisms.

Draft Genome Sequences of Two Toxigenic Corynebacterium ulcerans Strains.

Here, we present the draft genome sequences of two toxigenic Corynebacterium ulcerans strains isolated from two different patients: one from a blood sample and the other from a scar exudate following surgery. Although these two strains harbor the diphtheria toxin gene tox, no full prophage sequences were found in the flanking regions.

Carbapenem disks on MacConkey agar in screening methods for detection of carbapenem-resistant Gram-negative rods in stools.

Direct plating of simulated stool specimens on MacConkey agar (MCA) with 10-µg ertapenem, meropenem, and imipenem disks allowed the establishment of optimal zone diameters for the screening of carbapenem-resistant Gram-negative rods (CRGNR) of ≤ 24 mm (ertapenem), ≤ 34 mm (meropenem), and ≤ 32 mm (imipenem).

First Canadian outbreak of Enterobacteriaceae-expressing Klebsiella pneumoniae carbapenemase type 3.

BACKGROUND: Organisms expressing Klebsiella pneumoniae carbapenemase (KPC) are found in several regions worldwide but are rarely detected in Canada. The first outbreak of KPC-expressing strains of Enterobacteriaceae clinical isolates in a university-affiliated hospital intensive care unit (ICU) in Canada is described. METHODS: Enterobacteriaceae isolates that were flagged by the Vitek 2 (bioMérieux, France) system as possible carbapenemase producers were subjected to the modified Hodge test. Modified Hodge test-positive organisms were analyzed by pulsed-field gel electrophoresis, tested for KPC and other beta-lactamase genes by polymerase chain reaction analysis and underwent subsequent nucleic acid sequencing. Antimicrobial susceptibility profiles were determined by Vitek 2 and Etest (bioMérieux, France). A chart review was conducted to establish epidemiological links. RESULTS: During the study period, 10 unique Enterobacteriaceae isolates expressing KPC were detected from nine ICU patients. Five patients had infections (three pneumonias, one surgical site infection, one urinary tract infection). Isolates included Escherichia coli (5), Klebsiella oxytoca (2), Serratia marcescens (2) and Citrobacter freundii (1). Polymerase chain reaction analysis and sequencing confirmed the presence of KPC-3 in all isolates; four also carried TEM, two CTX-M and one CMY-2. The imipenem minimum inhibitory concentrations as determined by Etest ranged from 0.75 µg/mL to ≥32 µg/mL. Pulsed field gel electrophoresis clonal patterns and patient location in the ICU revealed presumptive horizontal transmission events. CONCLUSIONS: In the present study, Enterobacteriaceae isolates with KPC are emerging and can result in serious infections. The KPC gene can spread via plasmids to different genera of the Enterobacteriaceae family. The dissemination of KPC in Enterobacteriaceae and the consequences for treatment and infection control measures warrant a high degree of vigilance among clinicians and microbiologists.

HISTORIQUE: On trouve des organismes producteurs de carbapénèmases de type Klebsiella pneumoniae (KPC) dans diverses régions du monde, mais rarement au Canada. Les auteurs décrivent la première flambée de souches exprimant le KPC dans des isolats cliniques d'entérobactéries prélevés à l'unité de soins intensifs (USI) d'un hôpital universitaire du Canada. MÉTHODOLOGIE: Les chercheurs ont soumis au test de Hodge modifié les isolats d'entérobactéries que le système Vitek 2 (bioMérieux, France) signalait comme de possibles producteurs de carbapénèmases. Les chercheurs ont analysé les organismes positifs au test de Hodge par électrophorèse sur gel en champ pulsé, ont vérifié la présence de KPC et d'autres gènes de bêta-lactamase par analyse de la réaction en chaîne de la polymérase et ont ensuite effectué un séquençage de l'acide nucléique. Ils ont déterminé les profils de susceptibilité antimicrobienne par le système Vitek 2 et le test E (bioMérieux, France), puis procédé à une analyse des dossiers pour établir des liens épidémiologiques. RÉSULTATS: Pendant la période de l'étude, les chercheurs ont décelé dix isolats uniques d'entérobactéries productrices de KPC chez neuf patients de l'USI. Cinq patients avaient une infection (trois pneumonies, une infection au foyer d'une opération, une infection urinaire). Les isolats incluaient l'Escherichia coli (5), le Klebsiella oxytoca (2), le Serratia marcescens (2) et le Citrobacter freundii (1). L'analyse et le séquençage de la réaction en chaîne de la polymérase ont confirmé la présence de KPC-3 dans tous les isolats. Quatre contenaient un TEM, deux, un CTX-M, et un, un CMY-2. Les concentrations inhibitrices minimales de l'imipénem déterminées par test E variaient entre 0,75 µg/mL et au moins 32 µg/mL. Les schémas clonaux de l'électrophorèse en champ pulsé et la présence du patient à l'USI révélaient des événements présomptifs de transmission horizontale. CONCLUSIONS: Dans la présente étude, les isolats d'entérobactéries productrices de KPC sont émergents et peuvent donner lieu à de graves infections. Le gène de KPC peut se propager par les plasmides à divers genres de la famille des entérobactéries. La dissémination du KPC dans les entérobactéries et les conséquences pour les traitements et les mesures de contrôle des infections justifient un degré élevé de vigilance chez les cliniciens et les microbiologistes.