Evaluation of a Shotgun Metagenomics Approach for Detection of ESBL- and/or Carbapenemase-Producing Enterobacterales in Culture Negative Patients Recovered from Acute Leukemia.

Patients diagnosed with acute leukemia (AL) have a weakened immune system. Infections acquired by these patients are cause for concern and especially worrisome when Gram-negative multidrug-resistant (MDR) bacteria are involved, as they are difficult to treat, especially in the case of ESBL- and/or carbapenemase-producing Enterobacterales. Culture-based approaches have been relied on over the past decades as the method of choice for the early detection of gut colonization by MDR Gram-negative bacteria. However, various studies have indicated its limited sensitivity, underlining the need for new screening procedures in onco-hematological patients. Here, we evaluated a shotgun metagenomics approach to detect ESBL- and/or carbapenemase-producing Enterobacterales in the gut of 28 patients who had recovered from AL, which were previously colonized by these bacteria but cured at the time of sampling, as judged by culture-based methods. No ESBL or carbapenemase determinants were detected among the many resistance genes found by the metagenomics approach, supporting that patients were truly decolonized, with considerable consequences for their future clinical management. Due to the relatively low number of patients available for the present investigation, further studies should be conducted to support the utility and applicability of metagenomics for the routine screening of MDR bacteria in onco-hematological patients.

Clinical and Microbiological Risk Factors for 30-Day Mortality of Bloodstream Infections Caused by OXA-48-Producing Klebsiella pneumoniae.

Bloodstream infections (BSI) caused by carbapenem-resistant Klebsiella pneumoniae are associated with high morbidity and mortality, and the therapy options available for their treatment are frequently scarce. The aim of this study was to analyze risk factors for 30-day mortality in patients with BSI caused by OXA-48-producing K. pneumoniae. The clinical and treatment features of the patients, who attended a single hospital over a five-year period, were retrospectively reviewed. The microbiological features, including the sequence types (ST) and the somatic (O) and capsular (K) antigens, as well as their resistance properties, comprising phenotypes and genetic background, were also considered. To identify the risk factors for 30-day mortality, uni- and multivariate statistical analyses were performed. The univariate analysis revealed statistically significant correlations for age, male gender, lower respiratory system infection, infection by ST147 isolates, and infection by isolates expressing the K64 antigen. The multivariate analysis, applied to variables yielding p-values close to or lower than 0.05 in the univariate analysis, confirmed gender, lower respiratory system infection, and infection with ST147 isolates, but not age or infection with K64 isolates, as risk factors for 30-day mortality. Moreover, the multivariate analysis showed that patients suffering from hematological malignancies or having been treated with inappropriate therapy, both having p-values slightly higher than 0.05 in the univariate analysis, exhibited significantly poorer outcomes in the multivariant analysis. The association of the ST147 clone with an increased risk of mortality is a novel finding that deserves further attention. Studies like the one presented here can certainly benefit the management of patients with nosocomial BSI caused by carbapenemase-producing K. pneumoniae.

Detection of the optrA Gene Among Polyclonal Linezolid-Susceptible Isolates of Enterococcus faecalis Recovered from Community Patients.

Dispersion of transferable oxazolidinone resistance genes among enterococci poses a serious problem to human health. Prompt detection of bacteria carrying these genes is crucial to avoid their spread to multidrug-resistant bacteria. The aim of the study was to describe the presence of optrA-positive isolates among enterococci in a Spanish hospital, and to determine their genetic context and location through whole genome sequencing. All enterococci recovered in a Spanish hospital (Hospital El Bierzo; HEB) from February to December 2018 (n = 443), with minimal inhibitory concentrations (MICs) to linezolid (LZD) ≥4 mg/L, were tested by polymerase chain reaction for the presence of cfr, optrA, and poxtA transferable genes. Only four Enterococcus faecalis isolates (0.9%) had LZD MICs ≥4 mg/L and none of them was positive for cfr or poxtA genes. However, the optrA gene was detected in three isolates collected from urine samples of community patients, whose genomes were sequenced and subjected to bioinformatics analysis. These isolates belonged to different clones: ST7, ST480, and ST585. In these three isolates, the optrA gene was located on plasmids, associated with IS1216 in different arrays. In one isolate, the optrA plasmid coexists with a second plasmid, which carried multiple resistance genes for different classes of antibiotics. Detection of optrA-positive E. faecalis isolates in the community is a matter of concern. The spread of these bacteria into hospital settings, particularly in those, such as the HEB, where vancomycin-resistant enterococci are endemic, should be avoided, to preserve the efficacy of the last-resort oxazolidinones.

Plasmid-Mediated Quinolone Resistance (PMQR) in Two Clinical Strains of Salmonella enterica Serovar Corvallis.

Non-typhoid serovars of Salmonella enterica are one of the main causes of bacterial food-borne infections worldwide. For the treatment of severe cases of salmonellosis in adults, fluoroquinolones are amongst the drugs of choice. They are categorized by the World Health Organization (WHO) as "critically important with highest priority in human medicine". In the present study, two clinical S. enterica serovar Corvallis isolates (HUA 5/18 and HUA 6/18) from a Spanish hospital, selected on the basis of fluoroquinolone resistance, were characterized. The MICs of ciprofloxacin, determined by E-test, were 0.5 and 0.75 µg/mL for HUA 5/18 and HUA 6/18, respectively, and both were also resistant to pefloxacin but susceptible to nalidixic acid. Whole genome sequencing (WGS) of the isolates was performed with Illumina platform, and different bioinformatics tools were used for sequence analysis. The two isolates belonged to ST1541, and had the Thr57Ser substitution in the ParC protein which is also found in ciprofloxacin susceptible isolates. However, they harbored identical ColE plasmids of 10 kb carrying the qnrS1 gene. In these plasmids, the gene was flanked by defective versions of IS2-like and ISKra4-like insertion sequences. HUA 5/18 and HUA 6/18 were also phenotypically resistant to streptomycin, sulfonamides and tetracycline, with the responsible genes: strA, strB, sul2 and tet(A) genes, being located on a IncQ1 plasmid. ColE plasmids with the qnrS1 gene are widely spread among multiple serovars of S. enterica from different samples and countries. These mobilizable plasmids are playing an important role in the worldwide spread of qnrS1. Thus, their detection in hospitals is a cause of concern which deserves further attention.

Nosocomial Pneumonia Caused in an Immunocompetent Patient by the Emergent Monophasic ST34 Variant of Salmonella enterica Serovar Typhimurium: Treatment-Associated Selection of Fluoroquinolone and Piperacillin/Tazobactam Resistance.

The present report describes an uncommon case of nosocomial pneumonia caused by Salmonellaenterica in an immunocompetent patient. The patient was admitted to ICU of a tertiary hospital due to low level of consciousness, aphasia and seizure episodes. Four days after hospitalization, he developed nosocomial pneumonia, which evolved into septic shock. Gram-negative bacilli were recovered from blood, tracheal aspirate and fecal samples of the patient. The isolates, which were identified as Salmonella enterica, proved to be resistant to ciprofloxacin, amoxicillin/clavulanic acid and piperacillin/tazobactam. Four months before, the same bacterial species was recovered from feces and blood cultures of the patient, admitted to the nephrology ward of the same hospital with diagnosis of gastroenteritis and acute renal failure. However, at that time, the isolates were susceptible to the above-mentioned antibiotics. Genome sequencing revealed that all isolates were closely related and belonged to the emergent ST34 monophasic variant of S. enterica serovar Typhimurium. Since the patient has received therapy with fluoroquinolones and amoxicillin/clavulanic acid, these results support treatment-associated selection of the acquired resistances. In conclusion, this case represents a paradigm of selective pressure leading to in vivo development of resistance to highly relevant antibiotics, including the piperacillin/tazobactam combination used for empirical management of severe infections at ICU.

Establishment and Persistence of Glycopeptide-Resistant Enterococcus faecium ST80 and ST117 Clones Within a Health Care Facility Located in a Low-Prevalence Geographical Region.

Vancomycin-resistant Enterococcus faecium (VREfm) is one of the most important nosocomial pathogens with limited therapeutic alternatives. In this study, we followed the trends of VREfm and E. faecium causing bloodstream infections (BSIs) in a Spanish hospital, from 2011 to 2020. During this period, 832 E. faecium strains were isolated and 121 (14.5%) were vancomycin resistant. Nineteen of 101 BSIs (18.8%) caused by E. faecium were due to VREfm. The number of BSI-producing E. faecium isolates increased significantly over the past 5 years, with the percentage of invasive VREfm isolates being substantially higher than the average values in Europe and especially in Spain (<3%). VREfm isolates recovered in 2018 (28) and BSI-producing isolates from 2019 (3) and 2020 (2) were molecularly characterized. All were positive for vanA and belonged to sequence type (ST) 80 (28) or ST117 (5), within clonal complex 17. The isolates were only susceptible to linezolid, although most of them were also susceptible (dose dependent) to daptomycin. We report for the first time the establishment and persistence of the VREfm ST80 and ST117 clones in a Spanish hospital. The spread and establishment of hospital-adapted, multidrug-resistant VREfm clones in health care settings are cause for concern and may precede an increment in the BSIs caused by them.

High-Level Carbapenem Resistance among OXA-48-Producing Klebsiellapneumoniae with Functional OmpK36 Alterations: Maintenance of Ceftazidime/Avibactam Susceptibility.

The aim of this work was to analyze outer membrane porin-encoding genes (ompK35 and ompK36) in a collection of OXA-48 producing Klebsiella pneumoniae, to assess the effect of porin alterations on the susceptibility to ceftazidime/avibactam, and to describe a screening methodology for phenotypic detection of OXA-48-producing K. pneumoniae with disrupted porins. Antimicrobial susceptibility was tested by Microscan and Etest. The genomes of 81 OXA-48-producing K. pneumoniae were sequenced. MLST, detection of antimicrobial resistance genes, and analysis of ompK35 and ompK36 were performed in silico. Tridimensional structures of the OmpK36 variants were assessed. Receiver operating characteristics curves were built to visualize the performance ability of a disk diffusion assay using carbapenems and cefoxitin to detect OmpK36 functional alterations. A wide variety of OmpK36 alterations were detected in 17 OXA-48-producing K. pneumoniae isolates. All displayed a high-level meropenem resistance (MIC ≥ 8 mg/L), and some belonged to high-risk clones, such as ST15 and ST147. Alterations in ompK35 were also observed, but they did not correlate with high-level meropenem resistance. All isolates were susceptible to ceftazidime/avibactam and porin alterations did not affect the MICs of the latter combination. Cefoxitin together with ertapenem/meropenem low inhibition zone diameters (equal or lower than 16 mm) could strongly suggest alterations affecting OmpK36 in OXA-48-producing K. pneumoniae. OXA-48-producing K. pneumoniae with porin disruptions are a cause of concern; ceftazidime/avibactam showed good in vitro activity against them, so this combination could be positioned as the choice therapy to combat the infections caused by this difficult-to-treat isolates.

High Prevalence and Diversity of Cephalosporin-Resistant Enterobacteriaceae Including Extraintestinal Pathogenic E. coli CC648 Lineage in Rural and Urban Dogs in Northwest Spain.

The aim of this work was to assess the prevalence of extended spectrum-ß-lactamase (ESBL)- and carbapenemase-producing Enterobacteriaceae in fecal samples recovered from rural and urban healthy dogs in Northwest Spain (Galicia) to identify potential high-risk clones and to molecularly characterize positive isolates regarding the genes coding for ESBL/pAmpC resistance and virulence. Thirty-five (19.6%) out of 179 dogs were positive for cephalosporin-resistant Enterobacteriaceae, including Escherichiacoli and Klebsiella pneumoniae (39 and three isolates, respectively). All the isolates were multidrug resistant, with high rates of resistance to different drugs, including ciprofloxacin (71.4%). A wide diversity of ESBL/pAmpC enzymes, as well as E. coli phylogroups (A, B1, C, D, E, F and clade I) were found. The eight isolates (20.5%) found to conform to the ExPEC status, belonged to clones O1:H45-clade I-ST770 (CH11-552), O18:H11-A-ST93-CC168 (CH11-neg), O23:H16-B1-ST453-CC86 (CH6-31), and O83:H42-F-ST1485-CC648 (CH231-58), with the latter also complying the uropathogenic (UPEC) status. The three K. pneumoniae recovered produced CTX-M-15 and belonged to the ST307, a clone previously reported in human clinical isolates. Our study highlights the potential role of both rural and urban dogs as a reservoir of high-risk Enterobacteriaceae clones, such as the CC648 of E. coli and antimicrobial resistance traits. Within a One-Health approach, their surveillance should be a priority in the fight against antimicrobial resistance.

A Plasmid-Encoded FetMP-Fls Iron Uptake System Confers Selective Advantages to Salmonella enterica Serovar Typhimurium in Growth under Iron-Restricted Conditions and for Infection of Mammalian Host Cells.

The resistance plasmid pUO-StVR2, derived from virulence plasmid pSLT, is widespread in clinical isolates of Salmonella enterica serovar Typhimurium recovered in Spain and other European countries. pUO-StVR2 carries several genes encoding a FetMP-Fls system, which could be involved in iron uptake. We therefore analyzed S. Typhimurium LSP 146/02, a clinical strain selected as representative of the isolates carrying the plasmid, and an otherwise isogenic mutant lacking four genes (fetMP-flsDA) of the fetMP-fls region. Growth curves and determination of the intracellular iron content under iron-restricted conditions demonstrated that deletion of these genes impairs iron acquisition. Thus, under these conditions, the mutant grew significantly worse than the wild-type strain, its iron content was significantly lower, and it was outcompeted by the wild-type strain in competition assays. Importantly, the strain lacking the fetMP-flsDA genes was less invasive in cultured epithelial HeLa cells and replicated poorly upon infection of RAW264.7 macrophages. The genes were introduced into S. Typhimurium ATCC 14028, which lacks the FetMP-Fls system, and this resulted in increased growth under iron limitation as well as an increased ability to multiply inside macrophages. These findings indicate that the FetMP-Fls iron acquisition system exceeds the benefits conferred by the other high-affinity iron uptake systems carried by ATCC 14028 and LSP 146/02. We proposed that effective iron acquisition by this system in conjunction with antimicrobial resistance encoded from the same plasmid have greatly contributed to the epidemic success of S. Typhimurium isolates harboring pUO-StVR2.

Analysis of the Degradation of Broad-Spectrum Cephalosporins by OXA-48-Producing Enterobacteriaceae Using MALDI-TOF MS.

The objective of the study was to evaluate the activity of OXA-48 against different broad-spectrum cephalosporins and to identify the reaction products by MALDI-TOF MS. The action of OXA-48 on cefotaxime, ceftazidime, and ceftriaxone was assessed by this method, using an Escherichia coli J53 transconjugant carrying only the ~62 Kb IncL plasmid containing the blaOXA-48 gene, and the same strain without any plasmid was included as a negative control. In addition, a collection of 17 clinical OXA-48-producing Enterobacteriaceae, which were susceptible to broad-spectrum cephalosporins, was evaluated. MALDI-TOF MS-based analysis of the E. coli transconjugant carrying the blaOXA-48-harboring plasmid, and also the clinical isolates, showed degradation of cefotaxime into two inactive compounds-decarboxylated and deacetylated cefotaxime (~370 Da) and deacetyl cefotaxime (~414 Da), both with the hydrolyzed beta-lactam ring. Reaction products were not obtained when the experiment was performed with ceftriaxone or ceftazidime. From a clinical point of view, our study supports the idea that the efficacy of cefotaxime against OXA-48-producing Enterobacteriaceae is doubtful, in contrast to ceftazidime and ceftriaxone which could be valid choices for treating infections caused by these bacteria. However, further clinical studies confirming this hypothesis are required.

Antimicrobial Drug Resistance and Molecular Typing of Salmonella enterica Serovar Rissen from Different Sources.

Salmonella enterica serovar Rissen is one of the most common serovars found in pigs and pork products in different countries, including Spain. However, information on the molecular bases of antimicrobial drug resistance and the population structure of Salmonella Rissen from different sources in Spain is limited. The present study focused on 84 isolates collected in Spain from pig and beef carcasses, foods and clinical samples associated with sporadic cases of gastroenteritis, and one outbreak. The majority of the isolates were resistant to tetracycline (73.8%), mainly conferred by tet(A). Resistances to streptomycin (aadA1-like, aadA2, and strAB), sulfonamides (sul1, sul2, and sul3), trimethoprim (dfrA1-like and dfrA12), ampicillin (blaTEM-1-like), and chloramphenicol (cmlA1-like) were also detected, with frequencies ranging from 12% to 20.2%. Most of the identified genes were carried by integrons, including three class 1 integrons of the sul1 type, a class 1 integron of the sul3 type, and the class 2 integron of Tn7. Two sul1 integrons, the sul3 integron, and the class 2 integron are first reported in Salmonella Rissen. Typing of the isolates with XbaI pulsed-field gel electrophoresis detected a major clone, which was circulating in humans and animals during the past decade, and was responsible for the outbreak. The obtained results are relevant for food safety and public health.

Antimicrobial resistance and virulence determinants in European Salmonella genomic island 1-positive Salmonella enterica isolates from different origins.

Salmonella genomic island 1 (SGI1) contains a multidrug resistance region conferring the ampicillin-chloramphenicol-streptomycin-sulfamethoxazole-tetracycline resistance phenotype encoded by bla(PSE-1), floR, aadA2, sul1, and tet(G). Its increasing spread via interbacterial transfer and the emergence of new variants are important public health concerns. We investigated the molecular properties of SGI1-carrying Salmonella enterica serovars selected from a European strain collection. A total of 38 strains belonging to S. enterica serovar Agona, S. enterica serovar Albany, S. enterica serovar Derby, S. enterica serovar Kentucky, S. enterica serovar Newport, S. enterica serovar Paratyphi B dT+, and S. enterica serovar Typhimurium, isolated between 2002 and 2006 in eight European countries from humans, animals, and food, were subjected to antimicrobial susceptibility testing, molecular typing methods (XbaI pulsed-field gel electrophoresis [PFGE], plasmid analysis, and multilocus variable-number tandem-repeat analysis [MLVA]), as well as detection of resistance and virulence determinants (PCR/sequencing and DNA microarray analysis). Typing experiments revealed wide heterogeneity inside the strain collection and even within serovars. PFGE analysis distinguished a total of 26 different patterns. In contrast, the characterization of the phenotypic and genotypic antimicrobial resistance revealed serovar-specific features. Apart from the classical SGI1 organization found in 61% of the strains, seven different variants were identified with antimicrobial resistance properties associated with SGI1-A (S. Derby), SGI1-C (S. Derby), SGI1-F (S. Albany), SGI1-L (S. Newport), SGI1-K (S. Kentucky), SGI1-M (S. Typhimurium), and, eventually, a novel variant similar to SGI1-C with additional gentamicin resistance encoded by aadB. Only minor serovar-specific differences among virulence patterns were detected. In conclusion, the SGI1 carriers exhibited pathogenetic backgrounds comparable to the ones published for susceptible isolates. However, because of their multidrug resistance, they may be more relevant in clinical settings.

Food poisoning and Staphylococcus aureus enterotoxins.

Staphylococcus aureus produces a wide variety of toxins including staphylococcal enterotoxins (SEs; SEA to SEE, SEG to SEI, SER to SET) with demonstrated emetic activity, and staphylococcal-like (SEl) proteins, which are not emetic in a primate model (SElL and SElQ) or have yet to be tested (SElJ, SElK, SElM to SElP, SElU, SElU2 and SElV). SEs and SEls have been traditionally subdivided into classical (SEA to SEE) and new (SEG to SElU2) types. All possess superantigenic activity and are encoded by accessory genetic elements, including plasmids, prophages, pathogenicity islands, vSa genomic islands, or by genes located next to the staphylococcal cassette chromosome (SCC) implicated in methicillin resistance. SEs are a major cause of food poisoning, which typically occurs after ingestion of different foods, particularly processed meat and dairy products, contaminated with S. aureus by improper handling and subsequent storage at elevated temperatures. Symptoms are of rapid onset and include nausea and violent vomiting, with or without diarrhea. The illness is usually self-limiting and only occasionally it is severe enough to warrant hospitalization. SEA is the most common cause of staphylococcal food poisoning worldwide, but the involvement of other classical SEs has been also demonstrated. Of the new SE/SEls, only SEH have clearly been associated with food poisoning. However, genes encoding novel SEs as well as SEls with untested emetic activity are widely represented in S. aureus, and their role in pathogenesis may be underestimated.

[Evolutionary engineering in Salmonella: emergence of hybrid virulence-resistance plasmids in non-typhoid serotypes]. / Ingeniería evolutiva en Salmonella: la emergencia de plásmidos híbridos de virulencia-resistencia a antimicrobianos en serotipos no tifoideos.

An example of evolutive engineering in bacterial pathogens is the emergence of hybrid virulence-resistance (VR) plasmids in Salmonella enterica, resulting from an association between antimicrobial resistance determinants and specific virulence plasmids of the S. typhimurium and S. choleraesuis serotypes. VR plasmids all possess the spv (Salmonella plasmid virulence) operon, which is involved in systemic infection; however, they differ in the presence of other virulence determinants and in the resistance gene profile. VR plasmids of S. typhimurium have been found in Europe, and show resistance regions with different levels of complexity that can include class 1 integrons and various transposons. VR plasmids of S. choleraesuis, detected in strains isolated in Taiwan, only confer resistance to ampicillin and sulfonamides. Both serotypes are zoonotic and the presence of hybrid VR plasmids may confer an adaptive advantage under certain conditions, resulting in bacterial strains that are more difficult to treat and have a higher epidemic potential.

Ingeniería evolutiva en Salmonella: la emergencia de plásmidos híbridos de virulencia-resistencia a antimicrobianos en serotipos no tifoideos / Evolutionary engineering in Salmonella: emergence of hybrid virulence-resistance plasmids in non-typhoid serotypes

Un ejemplo de ingeniería evolutiva en bacterias patógenas es la emergencia en Salmonella enterica de plásmidos híbridos que han surgido por asociación de determinantes de resistencia (R) a antimicrobianos con plásmidos de virulencia (V) específicos de los serotipos typhimurium y choleraesuis. Los plásmidos VR poseen en común el operón spv (Salmonella plasmid virulence, que interviene en la infección sistémica) aunque difieren en la presencia de otros determinantes V y en el perfil de genes R. Los del serotipo typhimurium se han encontrado en Europa, y presentan regiones R con diferente grado de complejidad, pudiendo incluir integrones de clase 1 y distintos transposones. Los del serotipo choleraesuis proceden de cepas aisladas en Taiwán y sólo confieren resistencia a ampicilina y sulfonamidas. Ambos serotipos son zoonóticos y la formación de plásmidos VR puede aportar una ventaja selectiva en determinadas circunstancias, originando cepas más difíciles de tratar y con mayor potencial epidémico (AU)

An example of evolutive engineering in bacterial pathogens is the emergence of hybrid virulence-resistance (VR) plasmids in Salmonella enterica, resulting from an association between antimicrobial resistance determinants and specific virulence plasmids of the S. typhimurium and S. choleraesuis serotypes. VR plasmids all possess the spv (Salmonella plasmid virulence) operon, which is involved in systemic infection; however, they differ in the presence of other virulence determinants and in the resistance gene profile. VR plasmids of S. typhimurium have been found in Europe, and show resistance regions with different levels of complexity that can include class 1 integrons and various transposons. VR plasmids of S. choleraesuis, detected in strains isolated in Taiwan, only confer resistance to ampicillin and sulfonamides. Both serotypes are zoonotic and the presence of hybrid VR plasmids may confer an adaptive advantage under certain conditions, resulting in bacterial strains that are more difficult to treat and have a higher epidemic potential (AU)

[Outbreak of gastroenteritis in a nursery school caused by a strain of Salmonella enterica serovar Typhimurium carrying the hybrid virulence-resistance plasmid pUO-StVR2]. / Brote de gastroenteritis en una guardería causado por una cepa de Salmonella enterica serovar Typhimurium portadora del plásmido híbrido de resistencia-virulencia pUO-StVR2.

OBJECTIVE: Epidemiological and microbiological study of a salmonellosis outbreak, affecting 22 children in a nursery school in Oviedo (Spain). METHODS: Attack rates and epidemic curves were determined, and bacterial typing methods were applied. RESULTS: The outbreak was attributed to a Salmonella enterica serovar Typhimurium strain, belonging to an emergent type characterized by the presence of a hybrid virulence-resistance plasmid of 125-130 kb, named pUO-StVR2. The attack rate of confirmed cases vs. possible cases was 27.2% vs. 23.5% for the children and 0 vs. 26.5% for the staff of the affected center. The source of the infection could not be identified. Nevertheless, according to the evolution of the cases over time, the transmission route was likely to be personal contact between the staff and children, which facilitates fecal-oral dissemination. All but one of the 27 isolates analyzed (from 22 patients) showed identical features: R-profile, plasmid-profile, RAPD-type, PFGE-type; all were non-phage-typeable, with the exception of a DT104b isolate. pUO-StVR2 is probably a derivative of the virulence plasmid pSLT from the LT2 type strain that acquired an R-region complex (ACSSuT/blaOXA-catA1-strA/ B-aadA1-sul1-sul2-tet[B]), in which the blaOXA-aadA1 genes are part of the variable region of a class 1 integron. CONCLUSION: This outbreak is an example of how a Salmonella enterica serovar Typhimurium strain belonging to a type that is probably endemic in Spain can be transferred to the community and affect a susceptible population.

Brote de gastroenteritis en una guardería causado por una cepa de Salmonella enterica serovar Typhimurium portadora del plásmido híbrido de resistencia-virulencia pUO-StVR2 / Outbreak of gastroenteritis in a nursery school caused by a strain of Salmonella enterica serovar Typhimurium carrying the hybrid virulence-resistance plasmid pUO-StVR2

Objetivo. El estudio epidemiológico y microbiológico de un brote de salmonelosis ocurrido en 2004, que afectó a 22 niños de una guardería en Oviedo. Métodos. Se determinaron las tasas de ataque y curvas epidémicas y se aplicaron técnicas de tipificación bacteriana. Resultados. El brote pudo atribuirse a una cepa de Salmonella enterica serovar Typhimurium perteneciente a un grupo emergente que alberga un plásmido híbrido de virulencia-resistencia de 125-130 kb, denominado pUO-StVR2. Las tasas de ataque para los casos confirmados frente a los posibles fue 27,2% frente a 23,5% para los niños y 0 frente a 26,5% para el personal del centro. La fuente de infección no pudo ser identificada, aunque según la evolución temporal de los casos se podría considerar que el modo de transmisión fue el contacto cuidadoras-niños, facilitando la diseminación fecal-oral. Todos menos uno de los 27 aislamientos analizados (de 22 pacientes) presentaban características idénticas: perfil-R, perfil de plásmidos, RAPD-tipo, PFGE-tipo y eran no fagotipificables (la excepción fue un aislamiento con fagotipo DT104b). pUO-StVR2 es un derivado del plásmido de virulencia, pSLT, de la cepa tipo LT2 que ha ganado una región-R compleja (ACSSuT/blaOXA-catA1-strA/ B-aadA1-sul1-sul2-tet[B]) en la que los genes blaOXA-aadA1 forman parte de la región variable de un integrón de clase 1. Conclusión. Este brote constituye un ejemplo de cómo una cepa de S. enterica serovar Typhimurium, perteneciente a un tipo probablemente ya endémico en la península Ibérica, puede transmitirse a la comunidad y afectar a un colectivo susceptible (AU)

Objective. Epidemiological and microbiological study of a salmonellosis outbreak, affecting 22 children in a nursery school in Oviedo (Spain). Methods. Attack rates and epidemic curves were determined, and bacterial typing methods were applied. Results. The outbreak was attributed to a Salmonella enterica serovar Typhimurium strain, belonging to an emergent type characterized by the presence of a hybrid virulence-resistance plasmid of 125-130 kb, named pUO-StVR2. The attack rate of confirmed cases vs. possible cases was 27.2% vs. 23.5% for the children and 0 vs. 26.5% for the staff of the affected center. The source of the infection could not be identified. Nevertheless, according to the evolution of the cases over time, the transmission route was likely to be personal contact between the staff and children, which facilitates fecal-oral dissemination. All but one of the 27 isolates analyzed (from 22 patients) showed identical features: R-profile, plasmid-profile, RAPD-type, PFGE-type; all were non-phage-typeable, with the exception of a DT104b isolate. pUO-StVR2 is probably a derivative of the virulence plasmid pSLT from the LT2 type strain that acquired an R-region complex (ACSSuT/blaOXA-catA1-strA/ B-aadA1-sul1-sul2-tet[B]), in which the blaOXA-aadA1 genes are part of the variable region of a class 1 integron. Conclusion. This outbreak is an example of how a Salmonella enterica serovar Typhimurium strain belonging to a type that is probably endemic in Spain can be transferred to the community and affect a susceptible population (AU)