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1.
Clin Infect Dis ; 71(16): 2207-2210, 2020 11 19.
Artículo en Inglés | MEDLINE | ID: mdl-32147731

RESUMEN

We report diagnosis and management of the first laboratory-confirmed case of coronavirus disease 2019 (COVID-19) hospitalized in Toronto, Canada. No healthcare-associated transmission occurred. In the face of a potential pandemic of COVID-19, we suggest sustainable and scalable control measures developed based on lessons learned from severe acute respiratory syndrome.


Asunto(s)
Infecciones por Coronavirus , Coronavirus , Pandemias , Neumonía Viral , Coronavirus Relacionado al Síndrome Respiratorio Agudo Severo , Betacoronavirus , COVID-19 , Prueba de COVID-19 , Canadá , Técnicas de Laboratorio Clínico , Infecciones por Coronavirus/diagnóstico , Infecciones por Coronavirus/epidemiología , Humanos , Neumonía Viral/epidemiología , SARS-CoV-2
2.
J Antimicrob Chemother ; 73(suppl_7): vii5-vii11, 2018 07 01.
Artículo en Inglés | MEDLINE | ID: mdl-29982570

RESUMEN

Objectives: To assess antimicrobial susceptibility for 14 agents tested against 6001 invasive isolates of Streptococcus pneumoniae cultured from invasive patient samples from 2011 to 2015 as a part of the annual SAVE study. Methods: Isolates of S. pneumoniae were tested using the standard CLSI broth microdilution method (M07-A10, 2015) with MICs interpreted by CLSI M100 27th Edition (2017) MIC breakpoints. Results: From 2011 to 2015, small but significant increases (P ≤ 0.05) in the percentage susceptibility for penicillin (interpreted by all three CLSI MIC breakpoint criteria) (increase of 1.7%-3.2%), clindamycin (3.1%) and ceftriaxone (interpreted by non-meningitis and meningitis CLSI MIC breakpoint criteria) (1.1%-1.5%) were observed. Susceptibility rates for clarithromycin and other commonly tested antimicrobial agents remained unchanged (P > 0.05) over the 5 year period. Isolates with an MDR phenotype (resistance to three or more antimicrobial agent classes) decreased significantly (P < 0.001) from 8.5% in 2011 to 5.6% in 2015. Antimicrobial susceptibility rates were not generally associated (P > 0.05) with patient gender (exception: clarithromycin) but were associated (P ≤ 0.05) with patient age (chloramphenicol and clindamycin) or specimen source (penicillin, doxycycline, trimethoprim/sulfamethoxazole and clindamycin), as well as geographic location in Canada and concurrent resistance to penicillin or clarithromycin. Conclusions: The in vitro susceptibility of invasive isolates of S. pneumoniae in Canada to penicillin, clindamycin and ceftriaxone increased from 2011 to 2015, coincident with a significant decrease in MDR phenotypes.


Asunto(s)
Antibacterianos/farmacología , Farmacorresistencia Bacteriana Múltiple , Infecciones Neumocócicas/microbiología , Streptococcus pneumoniae/efectos de los fármacos , Adolescente , Adulto , Factores de Edad , Anciano , Canadá , Claritromicina/farmacología , Clindamicina/farmacología , Femenino , Humanos , Masculino , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Penicilinas/farmacología , Infecciones Neumocócicas/sangre , Infecciones del Sistema Respiratorio/microbiología , Factores de Riesgo , Adulto Joven
3.
J Antimicrob Chemother ; 73(suppl_7): vii12-vii19, 2018 07 01.
Artículo en Inglés | MEDLINE | ID: mdl-29982572

RESUMEN

Objectives: This study assessed MDR invasive isolates of Streptococcus pneumoniae, in relation to serotype evolution in Canada between 2011 and 2015 as part of the annual SAVE study. Methods: As part of a collaboration between the Canadian Antimicrobial Resistance Alliance and Public Health Agency of Canada-National Microbiology Laboratory, 6207 invasive isolates of S. pneumoniae were evaluated. All isolates were serotyped and had antimicrobial susceptibility testing performed, in accordance with CLSI guidelines (M07-A10, 2015). Complete susceptibility profiles were available for 6001 isolates. MDR was defined as resistance to three or more classes of antimicrobial agents (with penicillin MIC ≥2 mg/L defined as resistant). Results: The overall rate of MDR S. pneumoniae was 6.2% (372/6001) in SAVE, decreasing significantly from 8.5% in 2011 to 5.6% in 2015 (P = 0.0041). MDR was observed in 32 serotypes, with serotypes 15A and 19A predominating (26.6% and 41.7% of the MDR isolates, respectively). The overall proportion of serotypes 19A, 7F and 33A decreased significantly (P < 0.0001) throughout the study. The annual proportion of serotypes 7C, 8, 9N, 10A, 20, 24F, 29, 31, 33F, 35B and 38 increased throughout the study; however, among these increasing serotypes, MDR was only notable (>5%) for 24F and 33F. Conclusions: In 2015, 56.3% of invasive MDR S. pneumoniae were serotypes included in the PCV-13 vaccine. PCV-13 includes the most commonly identified serotype, 19A; however, other increasingly important MDR serotypes, such as 15A, 24F and 33F, are notably not in the currently used vaccines.


Asunto(s)
Antibacterianos/farmacología , Farmacorresistencia Bacteriana Múltiple , Streptococcus pneumoniae/efectos de los fármacos , Adolescente , Adulto , Anciano , Canadá , Niño , Preescolar , Claritromicina/farmacología , Humanos , Lactante , Recién Nacido , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Penicilinas/farmacología , Infecciones Neumocócicas/sangre , Infecciones Neumocócicas/microbiología , Vacunas Neumococicas/administración & dosificación , Serogrupo , Serotipificación , Streptococcus pneumoniae/clasificación , Vancomicina/farmacología , Adulto Joven
4.
J Antimicrob Chemother ; 73(suppl_7): vii20-vii31, 2018 07 01.
Artículo en Inglés | MEDLINE | ID: mdl-29982573

RESUMEN

Objectives: This study characterized the 11 most predominant serotypes of invasive Streptococcus pneumoniae infections collected by the annual SAVE study in Canada, between 2011 and 2015. Methods: A subset of the 11 most predominant serotypes (7F, 19A, 22F, 3, 12F, 11A, 9N, 8, 33F, 15A and 6C) collected by the SAVE study was analysed using PFGE and MLST, as well as PCR to identify pilus-encoding genes. WGS analyses were performed on a subset of the above isolates plus a random selection of background strains. Results: Of the predominant serotypes analysed, 7F, 33F and 19A were obtained more commonly from children <6 years of age, whereas 15A, 6C, 22F and 11A were more common in adults >65 years of age. Pneumococcal pilus PI-1 was identified in antimicrobial-susceptible serotype 15A (61/212) and <10% of 6C isolates (16/188). PI-2 was found in serotype 7F (683/701) and two-thirds of 11A isolates (162/241). Only serotype 19A-ST320 possessed both pili. Molecular and phylogenetic analyses identified serotypes 19A, 15A, 6C, 9N and 33F as highly diverse, whereas 7F, 22F and 11A demonstrated clonality. Antimicrobial resistance determinants were common within diverse serotypes, and usually similar within a clonal complex. Conclusions: Despite successful use of conjugate vaccines, S. pneumoniae remains a highly diverse organism in Canada. Several predominant serotypes, both antimicrobial susceptible and MDR, have demonstrated rapid clonal expansion or an increase in diversity. As S. pneumoniae continues to evolve in Canada, WGS will be a necessary component in the ongoing surveillance of antimicrobial-resistant and expanding clones.


Asunto(s)
Antibacterianos/farmacología , Infecciones Neumocócicas/epidemiología , Streptococcus pneumoniae/efectos de los fármacos , Streptococcus pneumoniae/genética , Adolescente , Adulto , Anciano , Técnicas de Tipificación Bacteriana , Canadá/epidemiología , Niño , Preescolar , Farmacorresistencia Bacteriana Múltiple/genética , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Tipificación de Secuencias Multilocus , Infecciones Neumocócicas/sangre , Infecciones Neumocócicas/microbiología , Reacción en Cadena de la Polimerasa , Serogrupo , Serotipificación , Secuenciación Completa del Genoma , Adulto Joven
5.
Appl Environ Microbiol ; 82(3): 939-53, 2016 02 01.
Artículo en Inglés | MEDLINE | ID: mdl-26590290

RESUMEN

A novel genomic island (LGI1) was discovered in Listeria monocytogenes isolates responsible for the deadliest listeriosis outbreak in Canada, in 2008. To investigate the functional role of LGI1, the outbreak strain 08-5578 was exposed to food chain-relevant stresses, and the expression of 16 LGI1 genes was measured. LGI1 genes with putative efflux (L. monocytogenes emrE [emrELm]), regulatory (lmo1851), and adhesion (sel1) functions were deleted, and the mutants were exposed to acid (HCl), cold (4°C), salt (10 to 20% NaCl), and quaternary ammonium-based sanitizers (QACs). Deletion of lmo1851 had no effect on the L. monocytogenes stress response, and deletion of sel1 did not influence Caco-2 and HeLa cell adherence/invasion, whereas deletion of emrE resulted in increased susceptibility to QACs (P < 0.05) but had no effect on the MICs of gentamicin, chloramphenicol, ciprofloxacin, erythromycin, tetracycline, acriflavine, and triclosan. In the presence of the QAC benzalkonium chloride (BAC; 5 µg/ml), 14/16 LGI1 genes were induced, and lmo1861 (putative repressor gene) was constitutively expressed at 4 °C, 37 °C, and 52 °C and in the presence of UV exposure (0 to 30 min). Following 1 h of exposure to BAC (10 µg/ml), upregulation of emrE (49.6-fold), lmo1851 (2.3-fold), lmo1861 (82.4-fold), and sigB (4.1-fold) occurred. Reserpine visibly suppressed the growth of the ΔemrELm strain, indicating that QAC tolerance is due at least partially to efflux activity. These data suggest that a minimal function of LGI1 is to increase the tolerance of L. monocytogenes to QACs via emrELm. Since QACs are commonly used in the food industry, there is a concern that L. monocytogenes strains possessing emrE will have an increased ability to survive this stress and thus to persist in food processing environments.


Asunto(s)
Antibacterianos/farmacología , Antiinfecciosos Locales/farmacología , Farmacorresistencia Bacteriana/genética , Genes MDR , Listeria monocytogenes/efectos de los fármacos , Listeria monocytogenes/genética , Compuestos de Amonio Cuaternario/farmacología , Acriflavina/farmacología , Proteínas Bacterianas/genética , Compuestos de Benzalconio/farmacología , Células CACO-2 , Canadá/epidemiología , Manipulación de Alimentos/normas , Industria de Procesamiento de Alimentos/normas , Islas Genómicas , Células HeLa , Humanos , Listeria monocytogenes/fisiología , Listeria monocytogenes/efectos de la radiación , Listeriosis/epidemiología , Listeriosis/microbiología , Listeriosis/prevención & control , Pruebas de Sensibilidad Microbiana , Mutación , Triclosán/farmacología
6.
J Clin Microbiol ; 53(1): 105-12, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25355764

RESUMEN

An efficient workflow to screen for and confirm the presence of carbapenemase-producing Gram-negative bacilli was developed by evaluating five chromogenic screening agar media and two confirmatory assays, the Rapid Carb screen test (Rosco Diagnostica A/S, Taastrup, Denmark) and the modified Hodge test. A panel of 150 isolates was used, including 49 carbapenemase-producing isolates representing a variety of ß-lactamase enzyme classes. An evaluation of analytical performance, assay cost, and turnaround time indicated that the preferred workflow (screening test followed by confirmatory testing) was the chromID Carba agar medium (bioMérieux, Marcy l'Étoile, France), followed by the Rapid Carb screen test, yielding a combined sensitivity of 89.8% and a specificity of 100%. As an optional component of the workflow, a determination of carbapenemase gene class via molecular means could be performed subsequent to confirmatory testing.


Asunto(s)
Antibacterianos/farmacología , Proteínas Bacterianas/genética , Bacterias Gramnegativas/efectos de los fármacos , Bacterias Gramnegativas/genética , Infecciones por Bacterias Gramnegativas/microbiología , Pruebas de Sensibilidad Microbiana/métodos , Juego de Reactivos para Diagnóstico , beta-Lactamasas/genética , Medios de Cultivo , Bacterias Gramnegativas/clasificación , Infecciones por Bacterias Gramnegativas/diagnóstico , Humanos , Reacción en Cadena de la Polimerasa Multiplex , Reproducibilidad de los Resultados , Flujo de Trabajo
7.
J Clin Microbiol ; 53(3): 951-60, 2015 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-25588655

RESUMEN

Shigellae cause significant diarrheal disease and mortality in humans, as there are approximately 163 million episodes of shigellosis and 1.1 million deaths annually. While significant strides have been made in the understanding of the pathogenesis, few studies on the genomic content of the Shigella species have been completed. The goal of this study was to characterize the genomic diversity of Shigella species through sequencing of 55 isolates representing members of each of the four Shigella species: S. flexneri, S. sonnei, S. boydii, and S. dysenteriae. Phylogeny inferred from 336 available Shigella and Escherichia coli genomes defined exclusive clades of Shigella; conserved genomic markers that can identify each clade were then identified. PCR assays were developed for each clade-specific marker, which was combined with an amplicon for the conserved Shigella invasion antigen, IpaH3, into a multiplex PCR assay. This assay demonstrated high specificity, correctly identifying 218 of 221 presumptive Shigella isolates, and sensitivity, by not identifying any of 151 diverse E. coli isolates incorrectly as Shigella. This new phylogenomics-based PCR assay represents a valuable tool for rapid typing of uncharacterized Shigella isolates and provides a framework that can be utilized for the identification of novel genomic markers from genomic data.


Asunto(s)
Disentería Bacilar/diagnóstico , Variación Genética , Genoma Bacteriano , Reacción en Cadena de la Polimerasa Multiplex/métodos , Filogenia , Shigella/clasificación , Shigella/aislamiento & purificación , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , Disentería Bacilar/microbiología , Humanos , Sensibilidad y Especificidad , Análisis de Secuencia de ADN , Shigella/genética
8.
J Antimicrob Chemother ; 70(7): 1960-4, 2015 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-25761605

RESUMEN

OBJECTIVES: Serotype replacement in Streptococcus pneumoniae following the implementation of a new vaccine has been associated with the emergence of non-vaccine serotypes as prominent causes of invasive pneumococcal disease (IPD). The aim of this study was to characterize specific non-PCV-13 serotypes 15A, 22F, 33F and 35B from IPD, isolated in Canada post-PCV-13 introduction in 2010. METHODS: Of 3802 IPD isolates collected from across Canada in 2011-13, 18.4% were found to be serotypes 15A, 22F, 33F and 35B. These 699 isolates were subjected to antimicrobial susceptibility testing, PFGE, MLST, molecular detection of pneumococcal pili and comparison with Pneumococcal Molecular Epidemiology Network (PMEN) clones. RESULTS: This study demonstrated clonal spread of specific STs, including MDR ST63 and its Sweden(15A)-25-related variants, the increasingly common ST433 and a variant of piliated, penicillin-non-susceptible ST558, related to PMEN clone Utah(35B)-24 (ST377). New STs of serotype 33F were identified. Several potential capsular switching events were identified within these serotypes. CONCLUSIONS: Non-PCV-13 serotype 22F is increasing in Canada through the rapid clonal expansion of ST433. Numerous new STs associated with serotype 33F indicate the potential divergence of the serotype. Serotypes 15A and 35B in Canada are related to international clones of S. pneumoniae.


Asunto(s)
Farmacorresistencia Bacteriana Múltiple , Genotipo , Infecciones Neumocócicas/microbiología , Serogrupo , Streptococcus pneumoniae/clasificación , Streptococcus pneumoniae/efectos de los fármacos , Factores de Virulencia/genética , Canadá/epidemiología , Proteínas Fimbrias/genética , Humanos , Pruebas de Sensibilidad Microbiana , Tipificación Molecular , Infecciones Neumocócicas/epidemiología , Vacunas Neumococicas/inmunología , Prevalencia , Streptococcus pneumoniae/aislamiento & purificación , Virulencia
9.
J Antimicrob Chemother ; 70(8): 2199-202, 2015 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-25921512

RESUMEN

OBJECTIVES: The goal of this study was to characterize Streptococcus pneumoniae demonstrating MDR (resistant to three or more antimicrobial classes) or XDR (resistant to five or more classes) phenotypes, collected from Canada during the CANWARD 2007-13 study. METHODS: From 2007 to 2013 inclusive, S. pneumoniae isolates were collected as a part of the CANWARD surveillance study. MDR and XDR isolates were subjected to PFGE, MLST, molecular detection of pneumococcal pili and macrolide resistance determinants mef(A/E) and erm(B), sequencing of PBPs 1A, 2B and 2X and comparison with Pneumococcal Molecular Epidemiology Network (PMEN) clones. RESULTS: Of 2129 S. pneumoniae isolates collected during the CANWARD 2007-13 study, 61 (2.9%) were found to be MDR. Of these MDR isolates, 43 (70.5%) were XDR. The most common serotypes for both MDR and XDR S. pneumoniae were 19A and 19F. Twenty-nine of 61 isolates (48%) demonstrated resistance to clarithromycin, clindamycin, doxycycline, penicillin and trimethoprim/sulfamethoxazole. All isolates possessed at least one macrolide resistance determinant and mutations in PBPs 1A, 2B and 2X. The most common clone was piliated, XDR ST320, an internationally circulating double-locus variant of Taiwan(19F)-14 (ST236). CONCLUSIONS: Though the rate of MDR S. pneumoniae has remained relatively stable since 2007, XDR strains have emerged in Canada. These strains are virulent, possess resistance determinants and are related to international clones.


Asunto(s)
Antibacterianos/farmacología , Farmacorresistencia Bacteriana Múltiple , Infecciones Neumocócicas/epidemiología , Infecciones Neumocócicas/microbiología , Streptococcus pneumoniae/efectos de los fármacos , Adolescente , Adulto , Canadá/epidemiología , Monitoreo Epidemiológico , Femenino , Genes Bacterianos , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Tipificación Molecular , Streptococcus pneumoniae/clasificación , Streptococcus pneumoniae/genética , Streptococcus pneumoniae/aislamiento & purificación , Adulto Joven
10.
Int J Syst Evol Microbiol ; 65(Pt 6): 1959-1966, 2015 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-25807976

RESUMEN

Polyphasic taxonomic analysis was performed on a clinical isolate (NML 06-3099T) from a cystic fibrosis patient, including whole-genome sequencing, proteomics, phenotypic testing, electron microscopy, chemotaxonomy and a clinical investigation. Comparative whole-genome sequence analysis and multilocus sequence analysis (MLSA) between Tatumella ptyseos ATCC 33301T and clinical isolate NML 06-3099T suggested that the clinical isolate was closely related to, but distinct from, the species T. ptyseos. By 16S rRNA gene sequencing, the clinical isolate shared 98.7 % sequence identity with T. ptyseos ATCC 33301T. A concatenate of six MLSA loci (totalling 4500 bp) revealed < 93.9 % identity between T. ptyseos ATCC 33301T, other members of the genus and the clinical isolate. A whole-genome sequence comparison between NML 06-3099T and ATCC 33301T determined that the average nucleotide identity was 76.24 %. The overall DNA G+C content of NML 06-3099T was 51.27 %, consistent with members of the genus Tatumella. By matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS analysis, NML 06-3099T had a genus-level match, but not a species-level match, to T. ptyseos. By shotgun proteomics, T. ptyseos ATCC 33301T and NML 06-3099T were found to have unique proteomes. The two strains had similar morphologies and multiple fimbriae, as observed by transmission electron microscopy, but were distinguishable by phenotypic testing. Cellular fatty acids found were typical for members of the Enterobacteriaceae. NML 06-3099T was susceptible to commonly used antibiotics. Based on these data, NML 06-3099T represents a novel species in the genus Tatumella, for which the name Tatumella saanichensis sp. nov. is proposed (type strain NML 06-3099T = CCUG 55408T = DSM 19846T).


Asunto(s)
Fibrosis Quística/microbiología , Enterobacteriaceae/clasificación , Filogenia , Adolescente , Técnicas de Tipificación Bacteriana , Composición de Base , Colombia Británica , ADN Bacteriano/genética , Enterobacteriaceae/genética , Enterobacteriaceae/aislamiento & purificación , Ácidos Grasos/química , Humanos , Masculino , Datos de Secuencia Molecular , Tipificación de Secuencias Multilocus , Hibridación de Ácido Nucleico/genética , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Esputo/microbiología
11.
J Clin Microbiol ; 52(4): 1081-8, 2014 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-24452166

RESUMEN

Vibrio parahaemolyticus is the leading bacterial cause of food-borne illness due to the consumption of contaminated seafood. The aim of the present study was to determine the population of its subtypes and establish a better understanding of the various types of V. parahaemolyticus strains that are causing human illness in Canada. The subtypes for 100 human clinical isolates of V. parahaemolyticus collected between 2000 and 2009 were determined by performing serotyping, ribotyping, pulsed-field gel electrophoresis, and multilocus sequence typing. Within this panel of strains, there was a high level of diversity (between 22 and 53 subtypes per method), but the presence of predominant clones with congruent subtypes between the various methods was also observed. For example, all 32 isolates belonging to sequence type 36 (ST36) were from serogroup O4, while 31 of them were ribotype EcoVib235-287, and 24 of the 32 were SfiI pulsed-field gel electrophoresis (PFGE) pattern VPSF1.0001. With regard to the presence of known virulence genes, 74 of the 100 isolates were PCR positive for the presence of the thermostable direct hemolysin (tdh); and 59 of these 74 strains also contained the second virulence marker, the tdh-related hemolysin (trh). The detection of trh was more predominant (81%) among the clinical isolates, and only four (4%) of the clinical isolates tested negative for the presence of both tdh and trh. This database, comprising 100 clinical isolates of V. parahaemolyticus strains from Canada, forms a baseline understanding of subtype diversity for future source attribution and other epidemiologic studies.


Asunto(s)
Tipificación Molecular , Serotipificación , Vibriosis/microbiología , Vibrio parahaemolyticus/clasificación , Vibrio parahaemolyticus/aislamiento & purificación , Canadá , Genotipo , Humanos , Fenotipo , Reacción en Cadena de la Polimerasa , Vibrio parahaemolyticus/genética , Vibrio parahaemolyticus/fisiología , Factores de Virulencia/genética
12.
BMC Microbiol ; 14: 70, 2014 Mar 19.
Artículo en Inglés | MEDLINE | ID: mdl-24641125

RESUMEN

BACKGROUND: The presence of Campylobacter jejuni temperate bacteriophages has increasingly been associated with specific biological effects. It has recently been demonstrated that the presence of the prophage CJIE1 is associated with increased adherence and invasion of C. jejuni isolates in cell culture assays. RESULTS: Quantitative comparative proteomics experiments were undertaken using three closely related isolates with CJIE1 and one isolate without CJIE1 to determine whether there was a corresponding difference in protein expression levels. Initial experiments indicated that about 2% of the total proteins characterized were expressed at different levels in isolates with or without the prophage. Some of these proteins regulated by the presence of CJIE1 were associated with virulence or regulatory functions. Additional experiments were conducted using C. jejuni isolates with and without CJIE1 grown on four different media: Mueller Hinton (MH) media containing blood; MH media containing 0.1% sodium deoxycholate, which is thought to result in increased expression of virulence proteins; MH media containing 2.5% Oxgall; and MHwithout additives. These experiments provided further evidence that CJIE1 affected protein expression, including virulence-associated proteins. They also demonstrated a general bile response involving a majority of the proteome and clearly showed the induction of almost all proteins known to be involved with iron acquisition. The data have been deposited to the ProteomeXchange with identifiers PXD000798, PXD000799, PXD000800, and PXD000801. CONCLUSION: The presence of the CJIE1 prophage was associated with differences in protein expression levels under different conditions. Further work is required to determine what genes are involved in causing this phenomenon.


Asunto(s)
Proteínas Bacterianas/biosíntesis , Ácidos y Sales Biliares/metabolismo , Campylobacter jejuni/metabolismo , Campylobacter jejuni/virología , Regulación Bacteriana de la Expresión Génica , Profagos/genética , Proteínas Bacterianas/genética , Medios de Cultivo/química , ADN Bacteriano/química , ADN Bacteriano/genética , Datos de Secuencia Molecular , Proteoma/análisis , Análisis de Secuencia de ADN
13.
Can J Microbiol ; 60(6): 417-8, 2014 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-24849211

RESUMEN

We report a concurrent case of infection with non-O157 Shiga-toxin-producing Escherichia coli (STEC) strain in an 8-month-old child. Laboratory and epidemiological investigations indicated child exposure to contaminated sheep meat following the Muslim feast of sacrifice (Eid al-Adha). Microbiological and molecular typing confirmed that the ovine strain O52:H45 (stx1+, eae-, hlyA-) was the causal agent. This is the first documented case of human infection to this STEC serotype.


Asunto(s)
Infecciones por Escherichia coli/microbiología , Carne/microbiología , Escherichia coli Shiga-Toxigénica/clasificación , Animales , Heces/microbiología , Humanos , Lactante , Masculino , Reacción en Cadena en Tiempo Real de la Polimerasa , Serotipificación , Ovinos , Toxina Shiga I/genética , Escherichia coli Shiga-Toxigénica/genética , Escherichia coli Shiga-Toxigénica/aislamiento & purificación
14.
Nat Rev Microbiol ; 22(9): 543-555, 2024 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-38789668

RESUMEN

Foodborne illnesses pose a substantial health and economic burden, presenting challenges in prevention due to the diverse microbial hazards that can enter and spread within food systems. Various factors, including natural, political and commercial drivers, influence food production and distribution. The risks of foodborne illness will continue to evolve in step with these drivers and with changes to food systems. For example, climate impacts on water availability for agriculture, changes in food sustainability targets and evolving customer preferences can all have an impact on the ecology of foodborne pathogens and the agrifood niches that can carry microorganisms. Whole-genome and metagenome sequencing, combined with microbial surveillance schemes and insights from the food system, can provide authorities and businesses with transformative information to address risks and implement new food safety interventions across the food chain. In this Review, we describe how genome-based approaches have advanced our understanding of the evolution and spread of enduring bacterial foodborne hazards as well as their role in identifying emerging foodborne hazards. Furthermore, foodborne hazards exist in complex microbial communities across the entire food chain, and consideration of these co-existing organisms is essential to understanding the entire ecology supporting pathogen persistence and transmission in an evolving food system.


Asunto(s)
Bacterias , Microbiología de Alimentos , Enfermedades Transmitidas por los Alimentos , Genoma Bacteriano , Enfermedades Transmitidas por los Alimentos/microbiología , Enfermedades Transmitidas por los Alimentos/epidemiología , Enfermedades Transmitidas por los Alimentos/prevención & control , Humanos , Bacterias/genética , Bacterias/clasificación , Inocuidad de los Alimentos
15.
BMC Med Genomics ; 17(1): 71, 2024 Mar 05.
Artículo en Inglés | MEDLINE | ID: mdl-38443925

RESUMEN

BACKGROUND: The timely and accurate diagnosis of bloodstream infection (BSI) is critical for patient management. With longstanding challenges for routine blood culture, metagenomics is a promising approach to rapidly provide sequence-based detection and characterisation of bloodborne bacteria. Long-read sequencing technologies have successfully supported the use of clinical metagenomics for syndromes such as respiratory illness, and modified approaches may address two requisite factors for metagenomics to be used as a BSI diagnostic: depletion of the high level of host DNA to then detect the low abundance of microbes in blood. METHODS: Blood samples from healthy donors were spiked with different concentrations of four prevalent causative species of BSI. All samples were then subjected to a modified saponin-based host DNA depletion protocol and optimised DNA extraction, whole genome amplification and debranching steps in preparation for sequencing, followed by bioinformatical analyses. Two related variants of the protocol are presented: 1mL of blood processed without bacterial enrichment, and 5mL of blood processed following a rapid bacterial enrichment protocol-SepsiPURE. RESULTS: After first identifying that a large proportion of host mitochondrial DNA remained, the host depletion process was optimised by increasing saponin concentration to 3% and scaling the reaction to allow more sample volume. Compared to non-depleted controls, the 3% saponin-based depletion protocol reduced the presence of host chromosomal and mitochondrial DNA < 106 and < 103 fold respectively. When the modified depletion method was further combined with a rapid bacterial enrichment method (SepsiPURE; with 5mL blood samples) the depletion of mitochondrial DNA improved by a further > 10X while also increasing detectable bacteria by > 10X. Parameters during DNA extraction, whole genome amplification and long-read sequencing were also adjusted, and subsequently amplicons were detected for each input bacterial species at each of the spiked concentrations, ranging from 50-100 colony forming units (CFU)/mL to 1-5 CFU/mL. CONCLUSION: In this proof-of-concept study, four prevalent BSI causative species were detected in under 12 h to species level (with antimicrobial resistance determinants) at concentrations relevant to clinical blood samples. The use of a rapid and precise metagenomic protocols has the potential to advance the diagnosis of BSI.


Asunto(s)
Saponinas , Sepsis , Humanos , ADN Mitocondrial , Metagenómica , Mitocondrias
16.
J Antimicrob Chemother ; 68(9): 1982-6, 2013 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-23710071

RESUMEN

OBJECTIVES: Over the last decade, a marked increase in Salmonella enterica serotype 4,[5],12:i:- with a core resistance to ampicillin, streptomycin, sulphonamides and tetracycline (ASSuT) has been observed in Europe. This study describes the emergence and characterization of isolates of multidrug-resistant Salmonella 4,[5],12:i:- in Canada. METHODS: Human clinical isolates of Salmonella 4,[5],12:i:- were identified by provincial laboratories from 2003 to 2010. Serotyping and phage typing were performed by standardized methodologies. MIC values were determined using broth microdilution. PCR was used to determine the presence of resistance genes. Multilocus sequence typing was performed on a selected number of isolates. RESULTS: A total of 26 251 Salmonella were submitted as part of the Canadian Integrated Program on Antibiotic Resistance Surveillance (CIPARS). Of these, Salmonella 4,[5],12:i:- accounted for a total of 766 isolates (2.9%), and the number increased significantly from 42 (1.4%) in 2003 to 164 (4.8%) in 2010. The ASSuT+ phenotype was observed in 11.9% (n = 91) of Salmonella 4,[5],12:i:- isolates and increased from two isolates in 2003 to 35 isolates in 2010. Two sequence types (STs) were observed. ST34 was mainly associated with the ASSuT isolates (n = 24; 38%), which contained blaTEM, strA-strB, tet(B) and sul2. ST19 was more likely to be associated with the ACSSuT phenotype and contained blaTEM, floR, strA-strB, sul2 and tet(A) or blaPSE-1, floR, aadA2, sul1 and tet(G). CONCLUSIONS: The prevalence of Salmonella 4,[5],12:i:- has significantly increased from 2003 to 2010 and it is now the fifth most common serotype reported in Canada causing human disease. Similar antimicrobial resistance patterns, phage types and STs have been observed in Europe.


Asunto(s)
Farmacorresistencia Bacteriana Múltiple , Monitoreo Epidemiológico , Infecciones por Salmonella/epidemiología , Infecciones por Salmonella/microbiología , Salmonella enterica/efectos de los fármacos , Salmonella enterica/aislamiento & purificación , Antibacterianos/farmacología , Tipificación de Bacteriófagos , Canadá/epidemiología , Genes Bacterianos , Genotipo , Humanos , Pruebas de Sensibilidad Microbiana , Tipificación de Secuencias Multilocus , Reacción en Cadena de la Polimerasa , Salmonella enterica/clasificación , Serotipificación
17.
Can J Microbiol ; 59(10): 671-8, 2013 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-24102220

RESUMEN

The monitoring of antimicrobial susceptibilities in Neisseria gonorrhoeae isolates and characterization of N. gonorrhoeae multiantigen sequence types (NG-MAST, ST) provide important surveillance data as resistance rates continue to rise. A total of 2970 N. gonorrhoeae isolates were collected by Canadian provincial public health laboratories in 2010, and 1233 were submitted to the National Microbiology Laboratory for testing. The NG-MAST and minimum inhibitory concentration (MIC) by agar dilution were determined for each isolate. Of the 2970 isolates, 25.1% were resistant to penicillin, 34.6% resistant to tetracycline, 31.5% resistant to erythromycin, 35.9% resistant to ciprofloxacin, and 1.2% resistant to azithromycin. Decreased susceptibility to cefixime (MIC ≥ 0.25 mg/L) and ceftriaxone (MIC ≥ 0.125 mg/L) was identified in 3.2% and 7.3% of the isolates, respectively. The most common STs found in Canada were ST1407 (13.3%), ST3150 (11.3%), and ST3158 (9.0%), with 249 different STs identified among the isolates. Within the ST1407 group, 19.5% and 43.3% isolates have decreased susceptibility to cefixime and ceftriaxone, respectively. ST1407, the most prevalent NG-MAST in Canada in 2010, has been associated with high-level ceftriaxone MICs and with cefixime treatment failure cases worldwide. Identification and monitoring of STs and corresponding antimicrobial resistance profiles may be useful in surveillance programs and be used to inform public health actions.


Asunto(s)
Antibacterianos/farmacología , Farmacorresistencia Bacteriana , Gonorrea/microbiología , Neisseria gonorrhoeae/efectos de los fármacos , Canadá/epidemiología , Monitoreo Epidemiológico , Gonorrea/epidemiología , Humanos , Pruebas de Sensibilidad Microbiana , Neisseria gonorrhoeae/genética , Neisseria gonorrhoeae/aislamiento & purificación , Prevalencia
18.
Can J Microbiol ; 59(12): 778-88, 2013 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-24313450

RESUMEN

The introduction of the 7-valent pneumococcal vaccine (PCV7) in Canada was very effective in reducing invasive pneumococcal disease (IPD) in children; however, increases of non-PCV7 serotypes have subsequently offset some of these reductions. A 13-valent pneumococcal vaccine (PCV13) targeting additional serotypes was implemented between 2010 and 2011, and in 2012 changes in the incidence of disease and the distribution of IPD serotypes began to emerge. The incidence of IPD in children <5 years of age declined from 18.0 to 14.2 cases per 100 000 population between 2010 and 2012; however, the incidence in ages ≥5 years remained relatively unchanged over the 3-year period, at about 9.7 cases per 100 000 population. From 2010 to 2012, PCV13 serotypes declined significantly from 66% (224/339) to 41% (101/244, p < 0.001) in children <5 years of age, and from 54% (1262/2360) to 43% (1006/2353, p < 0.001) in children ≥5 years of age. Serotypes 19A, 7F, 3, and 22F were the most common serotypes in 2012, with 19A decreasing from 19% (521/2727) to 14% (364/2620, p < 0.001), 7F decreasing from 14% (389/2727) to 12% (323/2620, p = 0.04), and 22F increasing from 7% (185/2727) to 11% (279/2620, p < 0.001) since 2010. Serotype 3 increased from 7% (23/339) to 10% (24/244) in <5-year-olds (p = 0.22) over the 3-year period. The highest rates of antimicrobial resistance were observed with clarithromycin (23%), penicillin using meningitis breakpoints (12%), clindamycin (8%), and trimethoprim-sulfamethoxazole (6%). Shifts in the distribution of IPD serotypes and reductions in the incidence of disease suggest that current immunization programs in Canada are effective in reducing the burden of IPD in children. While we acknowledge the limited data on the effectiveness of the PCV13 vaccine, to our knowledge, this study represents one of the first descriptions of the potential impact of the PCV13 vaccine in the Canadian population. Continued surveillance will be important to recognize replacement serotypes, to determine the extent of herd immunity effects in nonpaediatric populations, and to assess the overall effectiveness of PCV13 in reducing IPD in Canada.


Asunto(s)
Infecciones Neumocócicas/epidemiología , Infecciones Neumocócicas/prevención & control , Vacunas Neumococicas/administración & dosificación , Streptococcus pneumoniae/clasificación , Adolescente , Adulto , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Canadá/epidemiología , Niño , Preescolar , Femenino , Humanos , Incidencia , Lactante , Recién Nacido , Masculino , Persona de Mediana Edad , Infecciones Neumocócicas/microbiología , Serotipificación , Streptococcus pneumoniae/efectos de los fármacos , Streptococcus pneumoniae/aislamiento & purificación , Vacunación , Adulto Joven
19.
Food Microbiol ; 34(2): 319-27, 2013 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-23541198

RESUMEN

Antimicrobial resistance (AMR), co-selection phenomenon, and the relationship between reduced susceptibility (RSC) to ciprofloxacin (CIP) and resistance to other antimicrobials in Listeria spp. (n = 103) recovered from food processing environments (FPE) and food were investigated. Resistance of Listeria monocytogenes and other listeriae, respectively, to cefoxitin (FOX; 98% vs. 88%), CIP (7% vs. 4%), clindamycin (CLI; 33% vs. 59%) and tetracycline (6% vs. 8%) was observed, as was RSC to CIP (67% vs. 57%) and CLI (65% vs. 41%). L. monocytogenes also possessed RSC to linezolid (LZD; 6%), rifampicin (2%) and streptomycin (6%), with other listeriae displaying RSC to chloramphenicol (4%). L. monocytogenes serotype 1/2a (90%) isolates were more frequently resistant or possessed RSC to CIP compared to serotype 4b (55%) (p = 0.015). When eight strains were experimentally adapted to high concentrations of CIP, co-selection occurred as MICs to benzalkonium chloride (BAC) increased (n = 5), gentamicin MICs remained the same (n = 6) or increased 2-fold (n = 2), and led to RSC to LZD (n = 1) and resistance to CLI (n = 8). Overall, levels of resistance/RSC to CIP in food chain isolates, particularly 1/2a, are concerning. Further, reduced sensitivity to disparate antimicrobials following CIP exposure highlights the need for increased knowledge of co-selection phenomenon linked with antimicrobial agents.


Asunto(s)
Antibacterianos/farmacología , Productos Lácteos/microbiología , Farmacorresistencia Bacteriana Múltiple , Listeria/efectos de los fármacos , Alimentos Marinos/microbiología , Animales , Bovinos , Peces , Contaminación de Alimentos/análisis , Manipulación de Alimentos , Listeria/clasificación , Listeria/genética , Listeria/aislamiento & purificación
20.
Foodborne Pathog Dis ; 10(9): 747-56, 2013 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-23786604

RESUMEN

Most bacterial pathogens associated with human enteric illness have zoonotic origins and can be transmitted directly from animals to people or indirectly through food and water. This multitude of potential exposure routes and sources makes the epidemiology of these infectious agents complex. To better understand these illnesses and identify solutions to reduce human disease, an integrative approach like One Health is needed. This article considers the issue of Salmonella in Canada and interprets data collected by several Canadian surveillance and research programs. We describe recovery of Salmonella from various samples collected along the exposure pathway and compare the serovars detected in the different components under surveillance (animal, food, environment, and human). We then present three examples to illustrate how an approach that interprets multiple sources of surveillance data together is able to address issues that transcend multiple departments and jurisdictions. First, differences observed in recovery of Salmonella from different cuts of fresh chicken collected by different programs emphasize the importance of considering the surveillance objectives and how they may influence the information that is generated. Second, the high number of Salmonella Enteritidis cases in Canada is used to illustrate the importance of ongoing, concurrent surveillance of human cases and exposure sources to information domestic control and prevention strategies. Finally, changing patterns in the occurrence of ceftiofur-resistant Salmonella Heidelberg in retail meats and humans demonstrates how integrated surveillance can identify an issue in an exposure source and link it to a trend in human disease. Taken together, surveillance models that encompass different scales can leverage infrastructure, costs, and benefits and generate a multidimensional picture that can better inform disease prevention and control programs.


Asunto(s)
Monitoreo del Ambiente , Contaminación de Alimentos/prevención & control , Carne/microbiología , Salmonella enteritidis/aislamiento & purificación , Animales , Antibacterianos/farmacología , Canadá , Bovinos , Cefalosporinas/farmacología , Pollos , Farmacorresistencia Bacteriana Múltiple , Microbiología de Alimentos , Humanos , Pruebas de Sensibilidad Microbiana , Salmonella enteritidis/efectos de los fármacos , Porcinos
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