Resistance phenotype and virulence potential of Leclercia adecarboxylata strains isolated from different sources.

Introduction. Leclercia adecarboxylata is a member of Enterobacterales, often considered an opportunistic pathogen. Recent reports have highlighted L. adecarboxylata as an emerging pathogen harbouring virulence and resistance determinants.Gap statement. Little information exists on virulence and resistance determinants in L. adecarboxylata strains isolated from environmental, food, and clinical samples.Aim. To determine the presence of resistance and virulence determinants and plasmid features in L. adecarboxylata strains isolated from environmental, food, and clinical samples, as well as their phylogenetic relationship.Results. All strains tested showed resistance to ß-lactams and quinolones but were sensitive to aminoglycosides and nitrofurans. However, even though fosfomycin resistance is considered a characteristic trait of L. adecarboxylata, the resistance phenotype was only observed in 50 % of the strains; bla TEM was the most prevalent BLEE gene (70 %), while the quinolone qnrB gene was observed in 60 % of the strains. Virulence genes were differentially observed in the strains, with adhesion-related genes being the most abundant, followed by toxin genes. Finally, all strains carried one to seven plasmid bands ranging from 7 to 125 kbps and harboured several plasmid addiction systems, such as ParDE, VagCD, and CcdAB in 80 % of the strains.Conclusions. L. adecarboxylata is an important emerging pathogen that may harbour resistance and virulence genes. Additionally, it has mobilizable genetic elements that may contribute to the dissemination of genetic determinants to other bacterial genera.

Prior Carriage Predicts Intensive Care Unit Infections Caused by Extended-Spectrum Beta-Lactamase-Producing Enterobacteriaceae.

Intensive care unit-acquired infection (ICU-AI) and extended-spectrum beta-lactamase-producing Enterobacteriaceae (ESBL-PE) carriage are a major concern worldwide. Our objective was to investigate the impact of ESBL-PE carriage on ICU-AI. Our study was prospective, observational, and noninterventional. It was conducted over a 5-year period (Jan 2013-Dec 2017) in the medical-surgical intensive care unit of the Cayenne General Hospital (French Amazonia). During the study period, 1,340 patients were included, 271 (20.2%) developed ICU-AI, and 16.2% of these were caused by ESBL-PE. The main sites of ICU-AI were ventilator-associated pneumonia (35.8%) and primary bloodstream infection (29.8%). The main responsible microorganisms were Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae (ESBL-P in 35.8% of isolates), and Enterobacter cloacae (ESBL-P in 29.8% of isolates). Prior ESBL-PE carriage was diagnosed in 27.6% of patients with ICU-AI. In multivariable analysis, the sole factor associated with ESBL-PE as the responsible organism of ICU-AI was ESBL-PE carriage before ICU-AI (P < 0.001; odds ratio: 7.9 95% CI: 3.4-18.9). ESBL-PE carriers (74 patients) developed ICU-AI which was caused by ESBL-PE in 32 cases (43.2%). This proportion of patients carrying ESBL-PE who developed ICU-AI to the same microorganism was 51.2% in ESBL-P K. pneumoniae, 5.6% in ESBL-P Escherichia coli, and 40% in ESBL-P Enterobacter spp. NPV of ESBL-PE carriage to predict ICU-AI caused by ESBL-PE was above 94% and PPV was above 43%. Carriage of ESBL-P K pneumoniae and Enterobacter spp. is a strong predictor of ICU-AI caused by these two microorganisms.

Increased frequency of blaNDM in a tertiary care hospital in southern Brazil.

Resistance to carbapenems due to metallo-beta-lactamase NDM-1 was first described in Brazil in 2013. To date, only a few scattered reports of the prevalence of NDM-1 in the country have been reported, and most of them indicated a very low prevalence of this metalloenzyme. In the present study, we report a steady increase in the frequency of NDM among Enterobacterales resistant to carbapenems in a tertiary care hospital in southern Brazil. Carbapenemase genes were evaluated by multiplex real-time polymerase chain using high-resolution melting analysis among 3501 isolates of 8 different species of Enterobacterales recovered from January 2015 to May 2020. The blaKPC-like was identified in 3003 isolates (85.8%) and the blaNDM-like was the second most common gene (351 isolates-10%). There was a steady increase in frequency of blaNDM-like, from 4.2% in 2015 to 24% in 2020. The increase of blaNDM frequency raises an important matter as novel therapeutic options are currently very limited for the treatment of patients infected by bacteria carrying the blaNDM.

Evaluation of a polymyxin drop test for polymyxin resistance detection among non-fermentative gram-negative rods and enterobacterales resistant to carbapenems.

To assess the performance of the drop test for polymyxin B resistance detection among Enterobacterales and non-fermentative gram-negative rods resistant to carbapenems. Seven hundred and fifteen carbapenem-resistant isolates were tested: 628 Enterobacterales species and 87 non-fermentative gram-negative rods. For the polymyxin drop test, concentrations range from 0.25 to 8.0 µg/mL. Broth microdilution, as gold standard, was performed using in-house-prepared panels and interpreted according to the CLSI guidelines. Results were interpreted in terms of categorical agreements and discrepancies. Accuracy for a drop of polymyxin B at 2.0, 4.0 and 8.0 was calculated as better cutoff for resistance determination. No very major error was observed among all isolates, and 95.5% of agreement was observed among Enterobacterales, particularly for Klebsiella pneumoniae. A higher accuracy (95.1%) was obtained when a single drop of polymyxin B at 4.0 µg/mL was applied. Polymyxin drop test presented >95% of categorical agreement, without very major errors, for KPC-producing K. pneumoniae isolates. An accuracy of 95.1% was obtained with a single drop at 4.0 µg/mL polymyxin B. Polymyxin B drop is an easy and feasible test and may allow a reduction on the turnaround time for polymyxin resistance detection and impacting on early implementation of accurate therapeutic interventions.

Multidrug resistance and its association with Enterobacteriales and age among pregnant Peruvian women with- bacteremia.

INTRODUCTION: This study aimed to assess the prevalence of multidrug resistance (MDR) and its associated factors among pregnant Peruvian women with bacteremia. METHODOLOGY: In an 18-month cross-sectional study, all pregnant women were routinely tested with a presumptive diagnosis of sepsis admitted to the largest reference maternity hospital (Instituto Nacional Materno Perinatal) in Lima, Peru for bacteremia. Every isolate was tested for antimicrobial susceptibility as defined by the Institute of Clinical and Laboratory Standards (CLSI). Additionally, associated factors were assessed with MDR and the number of resistant antimicrobial categories using robust Poisson regression models with link log, especially focused on its association with age and bacterial families or species. RESULTS: A total of 236 blood cultures of pregnant women (33.4 ± 11.4 years old) was analyzed. The prevalence of MDR was 70% (95% confidence interval [CI]: 64%-76%). The main etiological agent was Escherichia coli (65%), showing an MDR rate of 74% (68%-81%). Overall, we observed that the MDR rate was associated with Enterobacteriales (adjusted prevalence rate, (aPR) = 1.29; 95% CI: 1.03-1.61) and age 35 or older (PR = 1.18; 95% CI: 1.01-1.39). However, the number of resistant antimicrobial categories was associated with Enterobacteriales (aPR = 1.44; 95% CI: 1.25-1.67) and hospital-acquired infections (PR = 0.81; 95% CI: 1.01-1.39). CONCLUSIONS: The prevalence of MDR among pregnant women with sepsis was alarmingly high, being even higher among women age 35 or older and among those with hospital-acquired infections.

Conjugative plasmidic AmpC detected in Escherichia coli, Proteus mirabilis and Klebsiella pneumoniae human clinical isolates from Portugal.

AmpC is a type of ß-lactamase enzyme produced by bacteria; these enzymes are classified in Class C and Group 1, and these confer resistance to cephamycin. Enterobacterales producing AmpC are reported worldwide and have great clinical importance due to therapeutic restriction and epidemiological importance once the easy dissemination by plasmidic genes to other bacteria is a real threat. These genes are naturally found in some enterobacteria as Enterobacter cloacae, Morganella morganii, and Citrobacter freundii, but other species have demonstrated similar resistance phenotype of AmpC production. Genes carried in plasmids have been described in these species conferring resistance to cefoxitin and causing therapeutic failure in some bacterial infections. This work detected and described five clinical strains of Escherichia coli, Proteus mirabilis, and Klebsiella pneumoniae that presented plasmid ampC (pAmpC) isolated from the north of Portugal collected in 2009. AmpC production was confirmed by inhibition of the enzyme by cloxacillin and boronic acid in agar diffusion tests. Also, PCR (polymerase chain reaction) was performed for the detection of gene universal to AmpC, blaampC, and others to AmpC group: blaACC, blaCIT, blaCMY, blaDHA, and blaEBC. The conjugation in liquid medium for 24 h was realized to determine if gene is localized in chromosome or plasmid. The isolates and their conjugants showed phenotypic characteristics and blaCMY and blaCIT were detected by PCR corroborating the AmpC characteristics observed in these bacteria. Confirmation of transfer of plasmid containing genes encoding AmpC is of high epidemiological relevance to the hospital studied and demonstrated the importance of AmpC surveillance and studies in hospital and community environments in order to choose the appropriate therapy for bacterial infections.

Detection of carbapenemase-producing bacteria in a public healthcare center from Venezuela.

INTRODUCTION: The dramatic increase in the prevalence and clinical impact of infections caused by Carbapenemase-Producing Bacteria in the nosocomial setting in Latin America represents an emerging challenge to public health. The present study detected carbapenemase-producing Gram-negative bacteria in patients from a Hospital from Venezuela, by phenotypic and genotypic methods. METHODOLOGY: The bacterial identification was carried out using conventional methods. The resistance to carbapenems was performed by Kirby-Baüer disk diffusion method, according to CLSI recommendations. The modified Hodge Test, double-disk with phenylboronic acid, double-disk with EDTA and Blue Carba Test were performed to detect phenotypic carbapenemase producers. The carbapenemase-encoding genes blaKPC, blaVIM, blaIMP, blaOXA-2, blaOXA-3, blaOXA-15 and blaOXA-21 were determined. RESULTS: The bacterial species identified were Klebsiella pneumoniae complex (181), Pseudomonas aeruginosa (51), and Acinetobacter baumannii-calcoaceticus complex (119). KPC-type was detected in 40.17% of isolates and VIM-type in 14.53%. KPC-type gene was only identified in K. pneumoniae isolates (77.9%). VIM-type gene was identified in P. aeruginosa (86.27%) and K. pneumoniae isolates (3.87%). There was not detection of IMP-type and OXA-type genes. CONCLUSIONS: We found a predominance of K. pneumoniae KPC producers and a high rate of VIM-producing P. aeruginosa. The epidemiology of CPB in Venezuela is rapidly evolving, and enhanced surveillance and reporting are needed across the healthcare continuum.

Detection of enterobacteria in broiler carcasses for sale in the market / Pesquisa de enterobactérias em carcaças de frangos à venda no comércio

This study aimed to analyze the microbiological quality of broiler marketed in Teresina, PI. The study was performed in the from September to November 2018, to which three groups of broiler carcasses produced in the city of Teresina were analyzed: without refrigeration, labeling and inspection (n=8); originated from local production, refrigerated, labeled, and inspected (n=8); and originated from other states, frozen, packed, labeled, and inspected (n=8). The collected samples were analyzed regarding the determination of the Most Probable Number (NMP) of total and thermotolerant coliforms, Escherichia coli, and detection of Salmonella spp., besides the measurement of the temperature of the carcasses. The logarithmic means of thermotolerant and total coliforms in the broiler carcasses varied from 0.70 to 4.66 NMP/g and from 0.73 to 4.66 NMP/g, respectively. The bacteria E. coli and Salmonella spp. were also detected in samples with refrigeration and inspection (25 % and 12.5 %, respectively). As for the marketing temperature, only the broiler carcass samples which were chilled and frozen were within the standard of the Brazilian legislation. The results observed in this study indicate the need for improvement in the processing, handling, and storage of the broiler meat marketed in Teresina, PI.(AU)

Objetivou-se analisar a qualidade microbiológica de carcaças de frangos comercializadas em Teresina, PI. O estudo foi realizado no período de setembro a novembro de 2018, para o qual foram analisadas carcaças de frango produzidas no município de Teresina, sem refrigeração, rotulagem e inspeção (n=8); de produção local refrigeradas, embaladas com rotulagem e inspecionadas (n=8); e oriundas de outros estados congeladas, embaladas com rotulagem e inspecionadas (n=8). As amostras coletadas foram analisadas quanto à determinação do Número Mais Provável (NMP) de coliformes totais e termotolerantes, Escherichia colie pesquisa de Salmonellaspp., além de aferição da temperatura das carcaças. As médias logarítmicas de coliformes termotolerantes e totais nas carcaçasde frango analisadas variaram de 0,70 a 4,66 NMP/g e de 0,73 a 4,66 NMP/g, respectivamente. Também foram encontradas bactérias E. colie Salmonellaspp. em amostras com refrigeração e com inspeção (25 % e 12,5 %, respectivamente). Quanto à temperatura de comercialização apenas as amostras de carcaça de frango resfriadas e as congeladas mostraram-se dentro do padrão da legislação brasileira. Os resultados observados nesse estudo indicam a necessidade de melhoria no processamento, manipulação e armazenamentoda carne de frango comercializado em Teresina, PI.(AU)

Detection of enterobacteria in broiler carcasses for sale in the market / Pesquisa de enterobactérias em carcaças de frangos à venda no comércio

This study aimed to analyze the microbiological quality of broiler marketed in Teresina, PI. The study was performed in the from September to November 2018, to which three groups of broiler carcasses produced in the city of Teresina were analyzed: without refrigeration, labeling and inspection (n=8); originated from local production, refrigerated, labeled, and inspected (n=8); and originated from other states, frozen, packed, labeled, and inspected (n=8). The collected samples were analyzed regarding the determination of the Most Probable Number (NMP) of total and thermotolerant coliforms, Escherichia coli, and detection of Salmonella spp., besides the measurement of the temperature of the carcasses. The logarithmic means of thermotolerant and total coliforms in the broiler carcasses varied from 0.70 to 4.66 NMP/g and from 0.73 to 4.66 NMP/g, respectively. The bacteria E. coli and Salmonella spp. were also detected in samples with refrigeration and inspection (25 % and 12.5 %, respectively). As for the marketing temperature, only the broiler carcass samples which were chilled and frozen were within the standard of the Brazilian legislation. The results observed in this study indicate the need for improvement in the processing, handling, and storage of the broiler meat marketed in Teresina, PI.

Objetivou-se analisar a qualidade microbiológica de carcaças de frangos comercializadas em Teresina, PI. O estudo foi realizado no período de setembro a novembro de 2018, para o qual foram analisadas carcaças de frango produzidas no município de Teresina, sem refrigeração, rotulagem e inspeção (n=8); de produção local refrigeradas, embaladas com rotulagem e inspecionadas (n=8); e oriundas de outros estados congeladas, embaladas com rotulagem e inspecionadas (n=8). As amostras coletadas foram analisadas quanto à determinação do Número Mais Provável (NMP) de coliformes totais e termotolerantes, Escherichia colie pesquisa de Salmonellaspp., além de aferição da temperatura das carcaças. As médias logarítmicas de coliformes termotolerantes e totais nas carcaçasde frango analisadas variaram de 0,70 a 4,66 NMP/g e de 0,73 a 4,66 NMP/g, respectivamente. Também foram encontradas bactérias E. colie Salmonellaspp. em amostras com refrigeração e com inspeção (25 % e 12,5 %, respectivamente). Quanto à temperatura de comercialização apenas as amostras de carcaça de frango resfriadas e as congeladas mostraram-se dentro do padrão da legislação brasileira. Os resultados observados nesse estudo indicam a necessidade de melhoria no processamento, manipulação e armazenamentoda carne de frango comercializado em Teresina, PI.

[Prevalence of extended-spectrum beta-lactamases in enterobacteria of neonatal sepsis and associated factors]. / Prevalencia de ß-lactamasas de espectro extendido en enterobacterias causantes de sepsis neonatal y factores asociados.

BACKGROUND: Infections caused by extended-spectrum beta-lactamases enterobacteria (ESBL-EP) have implications for neonatal morbidity and mortality. AIM: To describe the prevalence of ESBL-EP in neonatal sepsis and associated factors. METHODS: A prospective cohort study was conducted from August 2016 to August 2017; newborn babies (NB) hospitalized in the Hospital Civil de Guadalajara "Dr. Juan I. Menchaca" were included. The ESBL-EP were investigated by double-disk synergy test and its association with clinical and demographic characteristics of the NB. RESULTS: A total of 1,501 hospitalized NB were studied, with an average gestational age of 36.3 weeks. They were diagnosed 196 neonatal sepsis events, the most frequent etiologies were enterobacteria (45.5%). Resistance to ampicilin was found in 88.8% and to broad spectrum cephalosporins in more than 42% of the strains; 22.9% of them were ESBL phenotype. Apgar ≤ 7 at five minutes of life (OR 4.6; 95% CI 1.47-14.6) and gestational age < 37 weeks (OR 5.4; 95% CI 1.04-27.) increase the risk. CONCLUSION: In enterobacteria that cause neonatal sepsis, 22.9% were EP-ESBL; infection was more likely in patients with Apgar ≤ 7 at five minutes of age and in preterm infants.

Isolation of five Enterobacteriaceae species harbouring blaNDM-1 and mcr-1 plasmids from a single paediatric patient.

In Argentina, NDM metallo-ß-lactamase was first reported in 2013. By now, it has disseminated throughout the country in diverse Gram negative bacteria. Here, we report the case of a paediatric patient that underwent a 1-year hospitalisation due to erythrodermic psoriasis in 2014 and received multiple antimicrobial treatments. During his stay, five isolates were obtained from rectal swabs (rs) or blood culture (bc) suspicious of carbapenemase production: a K. quasipneumoniae subsp. quasipneumoniae (rs), Citrobacter freundii (rs), Escherichia coli (bc), Enterobacter cloacae (rs), and a Serratia marcescens (bc). The isolates were studied with broth microdilution, biparental conjugation and plasmid and whole genome sequencing (Illumina). All isolates harboured an 138,998-bp type 1 IncC plasmid that carried blaNDM-1, bleMBL, blaCMY-6, rmtC, aac(6')-Ib, and sul1 resistance genes. Additionally, the blaNDM-plasmids contained ISKpn8 an insertion sequence previously described as associated only to blaKPC. One isolate, a colistin-resistant E. coli, also carried a mcr-1-containing an IncI2 plasmid, which did not harbour additional resistance. The whole genome of K. quasipneumoniae subsp. quasipneumoniae isolate was fully sequenced. This isolate harboured, additionally to blaNDM, three plasmid-mediated quinolone resistance genes: qnrB4, qnrB52 and aac(6')-Ib-cr1. The E. cloacae isolate also harboured qnrA1. These findings alert to the underestimated horizontal dissemination of multidrug-resistant plasmids limiting treatment options with last resort antimicrobials.

Prevalencia de β -lactamasas de espectro extendido en enterobacterias causantes de sepsis neonatal y factores asociados / Prevalence of extended-spectrum beta-lactamases in enterobacteria of neonatal sepsis and associated factors

Resumen Introducción: Las infecciones causadas por enterobacterias productoras de β-talactamasas de espectro extendido (EP-BLEE) tienen implicaciones sobre la morbilidad y mortalidad neonatal. Objetivo: Describir la prevalencia de EP-BLEE en sepsis neonatal y los factores asociados. Métodos: Estudio de cohorte prospectivo, desde agosto del 2016 a agosto del 2017. Se incluyeron recién nacidos (RNs) ingresados en el Hospital Civil de Guadalajara "Dr. Juan I. Menchaca". Mediante prueba de difusión de doble disco se indagó la presencia de EP-BLEE y su asociación con características clínicas y demográficas de los RNs. Resultados: Se estudiaron 1.501 RNs hospitalizados, con edad gestacional promedio de 36,3 semanas. Se diagnosticaron 196 eventos de sepsis neonatal, la etiología más frecuente fueron enterobacterias (45,5%); 88,8% demostraron resistencia a ampicilina y más de 42% a cefalosporinas de amplio espectro. El 22,9% presentó fenotipo BLEE positivo. Tener Apgar ≤ 7 a los cinco minutos de vida (OR 4,6; IC 95% 1,47-14,6) y edad gestacional < 37 semanas (OR 5,4; IC 95%1,04-27,7) incrementaron el riesgo. Conclusión: En las enterobacterias causantes de sepsis neonatal, 22,9% son EP-BLEE; la infección es más probable en pacientes con Apgar ≤ 7 a los cinco minutos de vida y en prematuros.

Background: Infections caused by extended-spectrum beta-lactamases enterobacteria (ESBL-EP) have implications for neonatal morbidity and mortality. Aim: To describe the prevalence of ESBL-EP in neonatal sepsis and associated factors. Methods: A prospective cohort study was conducted from August 2016 to August 2017; newborn babies (NB) hospitalized in the Hospital Civil de Guadalajara "Dr. Juan I. Menchaca" were included. The ESBL-EP were investigated by double-disk synergy test and its association with clinical and demographic characteristics of the NB. Results. A total of 1,501 hospitalized NB were studied, with an average gestational age of 36.3 weeks. They were diagnosed 196 neonatal sepsis events, the most frequent etiologies were enterobacteria (45.5%). Resistance to ampicilin was found in 88.8% and to broad spectrum cephalosporins in more than 42% of the strains; 22.9% of them were ESBL phenotype. Apgar ≤ 7 at five minutes of life (OR 4.6; 95% CI 1.47-14.6) and gestational age < 37 weeks (OR 5.4; 95% CI 1.04-27.) increase the risk. Conclusion: In enterobacteria that cause neonatal sepsis, 22.9% were EP-ESBL; infection was more likely in patients with Apgar ≤ 7 at five minutes of age and in preterm infants.

Bacteriological quality of bottled water obtained from Mexico City small water purification plants: Incidence and identification of potentially pathogenic nontuberculous mycobacteria species.

The aim of this study was to determine the bacteriological quality of bottled water samples obtained from small purification plants located in Mexico City and to identify potentially pathogenic nontuberculous mycobacteria (NTM) species found in these samples. All 111 samples analyzed were positive for aerobic mesophilic bacteria (AMB) and 46 (41.4%) did not comply with Mexico's Official Guidelines. Sixty-nine (62.1%) and 23 (20.7%) water samples were positive for total coliforms (TC) and fecal coliforms (FC), respectively. A total of 81 (72.9%) of the water samples exceeded the maximum allowed limit stipulated in the guideline. Thirty-three (29.7%) of the purified water samples were positive for NTM, being recovered a total of 40 isolates. These NTM isolates were identified using three molecular markers (hsp65, rrs and rpoB genes) which corresponded to the fast-growing mycobacteria M. chelonae (n = 12), M. porcinum (n = 8), M. senegalense (n = 5), M. abscessus (n = 4), M. septicum (n = 4), M. wolinskyi (n = 3), M. mucogenicum (n = 2), M. fortuitum (n = 1) and M. sp. (n = 1). In seven purified water samples, two different NTM species were isolated simultaneously. Overall, these results showed that most of the purified bottled water samples analyzed in this study had unsatisfactory microbiological quality and some harbored NTM associated with illness. Our data could hasten health authorities to intensify efforts in the routine monitoring of activities in the purified bottled water industry in order to supply safe and healthy water to the public.

Microbiology of genus Raoultella, clinical features and difficulties in its diagnosis / Microbiología del género Raoultella, características clínicas y dificultades para su diagnóstico

The genus Raoultella, which belongs to the Enterobacteriaceae family, encompasses gramnegative, oxidase-negative, aerobic, encapsulated and motionless bacilli. Currently, four species of this genus have been described: Raoultella terrigena (R. terrigena), Raoultella planticola (R. planticola), Raoultella ornithinolytica (R. ornithinolytica) and Raoultella electrica (R. electrica), out of which R. planticola and R. ornithinolytica are the most important because of their probable association as pathogenic agents. The incidence of this genus has been underestimated, given that detecting it is really difficult, since it is misidentified as Klebsiella spp. by using manual and automated conventional phenotypic methods. The rapid and correct detection has been increasing since the advent of mass spectrometers (MALDI-TOF MS), which have been used for the differentiation of Klebsiella spp. and Raoultella spp. Currently 80 cases of bacteremia secondary to Raoultella spp. have been identified with different sites of primary infection, which is why it is important to emphasize proper detection in hospitals and healthcare facilities.

El género Raoultella, perteneciente a la familia Enterobacteriaceae, engloba bacilos gram negativos, oxidasa negativos, aeróbicos, encapsulados e inmóviles. Actualmente se han descrito cuatro especies de este género: Raoultella terrigena (R. terrigena), Raoultella planticola (R. planticola), Raoultella ornithinolytica (R. ornithinolytica) y Raoultella electrica (R. electrica), entre las que la R. planticola y la R. ornithinolytica son las más importantes por su probable asociación como agentes patógenos. Se ha subestimado la incidencia del género por la dificultad que presenta su caracterización al ser mal identificado como Klebsiella spp., al usar métodos fenotípicos manuales y automatizados convencionales. La rápida y correcta detección de Raoultella spp. ha ido aumentando desde la llegada de espectrómetros de masas (MALDI-TOF MS), útiles en la diferenciación entre Klebsiella spp. y Raoultella spp. Se han identificado 80 casos de bacteremia por Raoultella spp., con infección primaria en diversos sitios, por lo que es importante hacer énfasis en su correcta detección en los hospitales y centros de atención médica.

Improved blood culture workflow for faster identification of KPC-producing Enterobacterales.

Carba-NP original report for blood cultures described the need of subculture and mechanical lysis before testing, reaching the turnaround time of approximately 4 hours for sample preparation. We tested 100 consecutive blood cultures positive for Gram-negative bacilli on the Gram stain from a large clinical laboratory. Bacterial pellets were prepared by centrifugation and submitted to Carba-NP and Blue-Carba tests and used further to prepare smears for Vitek MS. Results obtained with colonies grown on sheep blood agar using the same methodologies were used as the gold standard. Carbapenemase genes were confirmed by PCR and DNA sequencing. Vitek MS identified correctly 86% of the samples. Of note, 7% of the samples were incorrectly reported by the instrument as containing a single isolate. KPC-2 was the predominant carbapenemase detected. There was 100% concordance for both negative and positive results for Carba-NP. In contrast, for Blue-Carba the concordance for positive results was 92.8%, and 41% of strains negative for carbapenemases presented a yellowish color on control well turning the test non-interpretable. The turnaround time for sample preparation for preparing the pellet was 13 min, and no subculture or mechanical lysis is needed when detecting KPC production in Enterobacterales.

Decreased antibiotic susceptibility of Enterobacteriaceae causing community-acquired urinary tract infections in French Amazonia.

OBJECTIVES: Urinary tract infections (UTIs) have rarely been studied in the Amazon region. We aimed to describe the epidemiology of bacteria causing UTIs in French Guiana. PATIENTS AND METHODS: We performed a monocenter retrospective study of adults consulting at the emergency department of Cayenne Hospital in 2014 with a diagnosis of UTI. The bacterial species and resistance profile were described. RESULTS: Two-hundred-and-eighty-nine patients presenting with UTI were included: 82 (28.4%) presented with cystitis, 166 (57.4%) with pyelonephritis, and 41 (14.2%) with male UTI. E. coli was predominant (74.1%), and had decreased susceptibility to ampicillin, amoxicillin-clavulanic acid, fluoroquinolones, co-trimoxazole, and furans compared with data from metropolitan France. Extended-spectrum beta-lactamases (ESBL) was isolated in 3.1% of E. coli and 31.6% of K. pneumoniae. CONCLUSIONS: Antibiotic susceptibility in UTI is lower than reported in metropolitan France without evidence for an excessive consumption of antibiotics.

Enterobacteria in the 21st century: a review focused on taxonomic changes.

INTRODUCTION: Enterobacteria are the main group causing infections in humans. The aim of this review is to present the new genera and the taxonomic changes that the Enterobacteriacea family has experienced in recent years. METHODOLOGY: a systematic search of papers published in databases from January 2000 to July 2018 was done. Additionally, the bibliographic references of each document were reviewed and each paper citing the article was reviewed in search of clinical cases. RESULTS: Nineteen new genera of Enterobacteria have been described since 2000. The genera Yersinia, Morganella and Erwinia do not belong to the family Enterobacteriacea anymore. CONCLUSIONS: for an adequate clinical and epidemiological interpretation, it is advisable to update the libraries of the commercial systems used for the identification of the microorganisms, as well as to train the staff in the taxonomic changes of microorganisms.

[Susceptibility to first-generation Cephalosporins in Enterobacteriaceae isolated from urine culture according to CLSI and EUCAST breakpoints]. / Estudio de susceptibilidad a cefalosporinas de primera generación en enterobacterias aisladas de urocultivo, según criterios CLSI y EUCAST.

Currently, the use of cefazolin is recommended to determine the susceptibility to first-generation oral cephalosporins in strains of enterobacteria in uncomplicated UTI. We determined susceptibility differences to oral cephalosporins in urinary strains according to cefazolin or cefalotin breakpoints and the correlation of susceptibility between cefazolin and cefadroxil. We studied 52 strains with cefalotin and cefazolin by disk-diffusion and MIC (Kirby-Bauer and Vitek XL) and a subgroup by disk-diffusion for cefadroxil. Agreement among different methods was 100% for K. pneumoniae and Proteus spp. In Escherichia coli, agreement for Vitek and disk-diffusion were 0 and 50% respectively. Susceptibility to first generation cephalosporins in E. coli should be determined with cefazolin. Agreement between cefazolin and cefadroxil suggests that cefazolin could also predict the susceptibility of cefadroxil.

Comparative genomics reveals diverse capsular polysaccharide synthesis gene clusters in emerging Raoultella planticola.

Raoultella planticola is an emerging zoonotic pathogen that is associated with rare but life-threatening cases of bacteremia, biliary tract infections, and urinary tract infections. Moreover, increasing antimicrobial resistance in the organism poses a potential threat to public health. In spite of its importance as a human pathogen, the genome of R. planticola remains largely unexplored and little is known about its virulence factors. Although lipopolysaccharides has been detected in R. planticola and implicated in the virulence in earlier studies, the genetic background is unknown. Here, we report the complete genome and comparative analysis of the multidrug-resistant clinical isolate R. planticola GODA. The complete genome sequence of R. planticola GODA was sequenced using single-molecule real-time DNA sequencing. Comparative genomic analysis reveals distinct capsular polysaccharide synthesis gene clusters in R. planticola GODA. In addition, we found bla TEM-57 and multiple transporters related to multidrug resistance. The availability of genomic data in open databases of this emerging zoonotic pathogen, in tandem with our comparative study, provides better understanding of R. planticola and the basis for future work.