Racial and Ethnic Disparities in Hospitalization and Clinical Outcomes Among Patients with COVID-19.

INTRODUCTION: The recent spread of coronavirus disease 2019 (COVID-19) has disproportionately impacted racial and ethnic minority groups; however, the impact of healthcare utilization on outcome disparities remains unexplored. Our study examines racial and ethnic disparities in hospitalization, medication usage, intensive care unit (ICU) admission and in-hospital mortality for COVID-19 patients. METHODS: In this retrospective cohort study, we analyzed data for adult patients within an integrated healthcare system in New York City between February 28-August 28, 2020, who had a lab-confirmed COVID-19 diagnosis. Primary outcome was likelihood of inpatient admission. Secondary outcomes were differences in medication administration, ICU admission, and in-hospital mortality. RESULTS: Of 4717 adult patients evaluated in the emergency department (ED), 3219 (68.2%) were admitted to an inpatient setting. Black patients were the largest group (29.1%), followed by Hispanic/Latinx (29.0%), White (22.9%), Asian (3.86%), and patients who reported "other" race-ethnicity (19.0%). After adjusting for demographic, clinical factors, time, and hospital site, Hispanic/Latinx patients had a significantly lower adjusted rate of admission compared to White patients (odds ratio [OR] 0.51; 95% confidence interval [CI] 0.34-0.76). Black (OR 0.60; 95% CI 0.43-0.84) and Asian patients (OR 0.47; 95% CI 0.25 - 0.89) were less likely to be admitted to the ICU. We observed higher rates of ICU admission (OR 2.96; 95% CI 1.43-6.15, and OR 1.83; 95% CI 1.26-2.65) and in-hospital mortality (OR 4.38; 95% CI 2.66-7.24; and OR 2.96; 95% CI 2.12-4.14) at two community-based academic affiliate sites relative to the primary academic site. CONCLUSION: Non-White patients accounted for a disproportionate share of COVID-19 patients seeking care in the ED but were less likely to be admitted. Hospitals serving the highest proportion of minority patients experienced the worst outcomes, even within an integrated health system with shared resources. Limited capacity during the COVID-19 pandemic likely exacerbated pre-existing health disparities across racial and ethnic minority groups.

Methicillin-Resistant Staphylococcus aureus Proteome Response to Antibiotic Stress Provides Insights for New Therapeutic Strategies.

Antimicrobial resistance is a global threat, with methicillin-resistant Staphylococcus aureus (MRSA) being one of the most representative drug-resistant pathogens. MRSA spread is increasing due to its ability to establish new reservoirs. To this end, the clonal complex (CC)-130 is an emerging genetic lineage, generally regarded as animal adapted and carrying the mecC gene, and sporadically found in humans. Although the MRSA antibiotic resistance mechanisms have been described, there are limited data on systems-wide omics responses to antibiotic stress, particularly at the proteome level. In this study, a gel-based quantitative proteomics approach was performed to assess the cellular responses of a mecC-harboring CC130 MRSA strain of human origin to subinhibitory doses of cefoxitin. We focused on the global response of MRSA to antibiotic stress and upon this treatment, 53 proteins were significantly differentially expressed. Most of the latter proteins were mapped to having functions in cellular metabolism while some glycolysis-related proteins showed a decreased expression after cefoxitin stress. On the contrary, pyruvate kinase, a potential antimicrobial drug target, was found upregulated. Also, quorum sensing, genetic information processing, and stress response proteins were found upregulated. Low-affinity penicillin-binding protein (mecC-encoded) was found in cefoxitin-treated samples. In conclusion, these new findings on cefoxitin-induced proteome changes provide important insights and molecular leads for innovation in treatment of MRSA specifically, and omics approaches to address antibiotic resistance generally.

Multidrug-resistant Klebsiella pneumoniae harboring extended spectrum ß-lactamase encoding genes isolated from human septicemias.

Klebsiella pneumoniae is a major pathogen implicated in nosocomial infections. Extended-spectrum ß-lactamase (ESBL)-producing K. pneumoniae isolates are a public health concern. We aim to characterize the type of ß-lactamases and the associated resistance mechanisms in ESBL-producing K. pneumoniae isolates obtained from blood cultures in a Portuguese hospital, as well as to determine the circulating clones. Twenty-two cefotaxime/ceftazidime-resistant (CTX/CAZR) K. pneumoniae isolates were included in the study. Identification was performed by MALDI-TOF MS and the antimicrobial susceptibility testing by disk-diffusion. The screening test for ESBL-production was performed and ESBL-producer isolates were further characterized. The presence of different beta-lactamase genes (blaCTX-M, blaSHV, blaTEM, blaKPC, blaNDM, blaVIM, blaOXA-48, blaCMY-2, blaDHA-1, blaFOX, blaMOX, and blaACC) was analyzed by PCR/sequencing in ESBL-producer isolates, as well as the presence of other resistance genes (aac(6')-Ib-cr, tetA/B, dfrA, qnrA/B/S, sul1/2/3) or integron-related genes (int1/2/3). Multilocus-sequence-typing (MLST) was performed for selected isolates. ESBL activity was detected in 12 of the 22 CTX/CAZR K. pneumoniae isolates and 11 of them carried the blaCTX-M-15 gene (together with blaTEM), and the remaining isolate carried the blaSHV-106 gene. All the blaCTX-M-15 harboring isolates also contained a blaSHV gene (blaSHV-1, blaSHV-11 or blaSHV-27 variants). Both blaSHV-27 and blaSHV-106 genes correspond to ESBL-variants. Two of the CTX-M-15 producing isolates carried a carbapenemase gene (blaKPC2/3 and blaOXA-48) and showed imipenem resistance. The majority of the ESBL-producing isolates carried the int1 gene, as well as sulphonamide-resistance genes (sul2 and/or sul3); the tetA gene was detected in all eight tetracycline-resistant isolates. Three different genetic lineages were found in selected isolates: ST348 (one CTX-M-15/TEM/SHV-27/KPC-2/3-producer isolate), ST11 (two CTX-M-15/TEM/SHV-1- and CTX-M-15-TEM-SHV-11-OXA-48-producer isolates) and ST15 (one SHV-106/TEM-producer isolate). ESBL enzymes of CTX-M-15 or SHV-type are detected among blood K. pneumoniae isolates, in some cases in association with carbapenemases of KPC or OXA-48 type.

Genetic characterization of extended-spectrum ß-lactamase-producing Enterobacteriaceae from a biological industrial wastewater treatment plant in Tunisia with detection of the colistin-resistance mcr-1 gene.

This study evaluated the occurrence of extended-spectrum ß-lactamases (ESBL) and associated resistance genes, integrons, and plasmid types, as well as the genetic relatedness of enterobacterial isolates in the wastewater treatment plant (WWTP) of La Charguia, Tunis City (Tunisia). A total of 100 water samples were collected at different points of the sewage treatment process during 2017-2019. Antimicrobial susceptibility was conducted by the disc-diffusion method. blaCTX-M, blaTEM and blaSHV genes as well as those encoding non-ß-lactam resistance, the plasmid types, occurrence of class1 integrons and phylogenetic groups of Escherichia coli isolates were determined by PCR/sequencing. Genomic relatedness was determined by multi-locus sequence typing (MLST) for selected isolates. In total, 57 ESBL-producer isolates were recovered (47 E. coli, eight Klebsiella pneumoniae, 1 of the Citrobacter freundii complex and 1 of the Enterobacter cloacae complex). The CTX-M-15 enzyme was the most frequently detected ESBL, followed by CTX-M-27, CTX-M-55 and SHV-12. One E. coli isolate harboured the mcr-1 gene. The following phylogroups/sequence types (STs) were identified among ESBL-producing E. coli isolates: B2/ST131 (subclade-C1), A/ST3221, A/ST8900, D/ST69, D/ST2142, D/ST38, B1/ST2460 and B1/ST6448. High numbers of isolates harboured the class 1 integrons with various gene cassette arrays as well as IncP-1 and IncFIB plasmids. Our findings confirm the importance of WWTPs as hotspot collectors of ESBL-producing Enterobacteriaceae with a high likelihood of spread to human and natural environments.

Penicillin-susceptible Staphylococcus aureus bacteremia: Epidemiological and clinical relevance. Possible therapeutic implications. / Bacteriemia por Staphylococcus aureus sensible a penicilina. Importancia epidemiológica, clínica y posibles implicaciones terapéuticas.

INTRODUCTION: The increase in penicillin susceptibility among Staphylococcus aureus (SA-PenS) might have therapeutic relevance. We aimed to study the current situation in our environment. MATERIAL AND METHODS: Over a 2.5 years period, all SA isolates from bacteraemia in one hospital were analysed. For all isolates, antimicrobial susceptibility profile, beta-lactam resistance genes (blaZ, mecA) and Panton-Valentine leucocidine encoding-genes were studied. For SA-PenS-blaZnegative isolates, spa-type, MLST and the presence of other resistance genes were studied. RESULTS: Among 84 patients with SA bacteraemia (35.7% MRSA and 64.3% MSSA), 77 were analysed; 22.2% of MSSA isolates were PenS and blaZnegative (Pen-MIC≤0.03µg/ml) corresponding to 14.3% of the total SA. In MSSA-PenS-blaZnegative isolates, eight spa-types and 7 clonal-complexes were detected. CONCLUSION: A high prevalence of MRSA/SA and MSSA-PenS-blaZnegative/MSSA was detected in blood cultures. Pen-MIC≤0,3µg/ml corresponded to MSSA-PenS-blaZnegative. This situation raises therapeutic options which should be further evaluated in larger studies and clinical trials.

Antimicrobial resistance phenotypes and genotypes of methicillin-resistant Staphylococcus aureus CC398 isolates from Spanish hospitals.

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) of lineage CC398 is an emerging clone causing human infections but is mostly found in pigs. The aim of this study was to characterize the antimicrobial resistance phenotypes/genotypes of a collection of 137 MRSA CC398 isolates obtained in a previous study from 17 Spanish hospitals, using tetracycline resistance as marker for selection. A multidrug-resistant (MDR) phenotype was present in 79% of analysed isolates, with 17% of them resistant to at least six different antimicrobial families. All tetracycline-resistant isolates (n=137) carried the tetM gene and 75% also carried the tetK gene. Almost 50% of MRSA CC398 isolates showed macrolide and/or lincosamide resistance: a) 39% of isolates were ERYR-CLIR (all with constitutive phenotype), with 87% of them carrying the ermC gene, followed by msrA (25%), ermB (21%), vgaA (17%), ermA (6%), lsaB (4%), linA (2%), linB (2%), and ermT (2%, this isolate with the new spa-type t18071); and b) 9% of MRSA CC398 isolates showed the dissociated ERYS-CLIR phenotype carrying the linA, linB, lsaB and vgaA genes. Other antimicrobial resistance phenotypes in these MRSA CC398 isolates included resistance to ciprofloxacin (67%), aminoglycosides (21%), mupirocin (6%), chloramphenicol (4%) or fusidic acid (2%). The more common resistance genes detected for some of these antimicrobials were: aac(6')-Ie-aph(2'')-Ia (16%) and ant(4')-Ia (12%) for aminoglycosides, and fexA (3%) for chloramphenicol. The high rate of MDR phenotypes with a wide range of antimicrobial resistance genes shown in this study reduce the potential therapeutic options in case of infections.

Caracterización de mecanismos de resistencia a carbapenémicos en aislados clínicos de Pseudomonas aeruginosa en un hospital español / Characterisation of carbapenem-resistance mechanisms in clinical Pseudomonas aeruginosa isolates recovered in a Spanish hospital

INTRODUCCIÓN: Los carbapenémicos son los antibióticos betalactámicos con mayor espectro de actividad en el tratamiento de infecciones por Pseudomonas aeruginosa. El objetivo de este trabajo fue caracterizar molecularmente una colección de aislados de P. aeruginosa resistentes a carbapenémicos (PARC). MÉTODOS: Se recogieron 85 aislados PARC de 60 pacientes en el Hospital San Pedro, Logroño (período 2008-2011). La relación clonal se determinó por electroforesis en gel de campo pulsado (PFGE), la sensibilidad a 15 antipseudomónicos por método de difusión en disco y las alteraciones en oprD, la caracterización de integrones y la tipificación molecular (MLST) por PCR y secuenciación. RESULTADOS: Las 85 PARC se clasificaron en 35 perfiles diferentes de PFGE. Se seleccionaron 61 cepas de los 60 pacientes y se observó que eran multirresistentes, aunque ninguna mostró fenotipo carbapenemasa. Se detectó un gran polimorfismo de OprD, destacando que el 59% de las cepas presentaban un codón de finalización prematuro. ISPa1328 e ISPsp4 truncaban el gen oprD en 2 cepas (GenBank KF517097 y KF517098). El 67% de las cepas presentó integrones de clase 1 con genes codificantes de enzimas modificantes de aminoglucósidos, 2 de las cuales portaban un nuevo integrón: aac(3)-Ia+aadA1h (nombrado In272, GenBank GQ144317). Se detectaron 4 secuencias tipo (ST) (número de cepas): ST175 (35), ST308 (3), ST235 (2) y ST639 (1). CONCLUSIÓN: La multirresistencia, el alto polimorfismo de oprD, el alto porcentaje de integrones, la moderada relación clonal de las cepas y la elevada diseminación epidémica de clones de alto riesgo son aspectos de gran preocupación clínica para erradicar la diseminación de PARC

INTRODUCTION: Carbapenems are the beta-lactam antibiotics with the best spectrum of activity in the treatment of Pseudomonas aeruginosa infections. The objective of this study was to molecularly characterise a collection of carbapenem-resistant P. aeruginosa isolates (PARC). METHODS: A total of 85 PARC isolates were recovered from 60 patients in the Hospital San Pedro, Logroño (period 2008-2011). Clonal relationship was determined using pulsed-field gel electrophoresis (PFGE), susceptibility testing to 15 anti-pseudomonal agents was performed using the disk diffusion method, and alterations in oprD, characterisation of integrons and molecular typing (MLST) using PCR and sequencing. RESULTS: The 85 PARC were classified into 35 different PFGE profiles. Of the 61 selected strains from 60 patients all of them were multiresistant, although none of them showed a carbapenemase phenotype. High polymorphism was detected in OprD, emphasising that 59% of the strains had a premature stop codon. ISPa1328 and ISPsp4 insertion sequences truncated oprD gene into 2 strains (GenBank KF517097 and KF517098). Two-thirds (67%) of the strains showed class 1 integrons with genes encoding aminoglycoside modifying enzymes, and 2 of them carried a new integron: aac(3)-Ia+aadA1h, named In272, GenBank GQ144317. Four sequence types were detected (Strain Nos.): ST175 (35), ST308 (3), ST235 (2), and ST639 (1). CONCLUSION: Multidrug resistance, high polymorphism in oprD, a high percentage of integrons, moderate clonal relationship of strains, and the high epidemic dissemination of high-risk clones are clinical aspects of great concern in order to eradicate the spread of PARC

Characteristics of ciprofloxacin-resistant Enterobacteriaceae isolates recovered from wastewater of an Algerian hospital.

INTRODUCTION: Hospital effluents are a source of environmental pollution by drugs, antibiotic-resistant bacteria, and resistance genes. Quinolones, particularly ciprofloxacin, are commonly detected in these effluents, contributing to the emergence of antimicrobial resistance. The objective of this study was to characterize ciprofloxacin-resistant Enterobacteriaceae in hospital effluents. METHODOLOGY: Isolates were selected on Tergitol-7 agar supplemented with ciprofloxacin and genotyped by ERIC-PCR. Antibiotic susceptibility testing was done using the disk diffusion method, and minimum inhibitory concentrations were determined using the agar dilution method. Resistance genes, integrons, phylogenetic groups, and sequence types were identified by PCR and sequencing. RESULTS: A total of 17 ciprofloxacin-resistant isolates were characterized: Escherichia coli, Escherichia vulneris, Klebsiella pneumoniae, Klebsiella oxytoca, Citrobacter freundii, and Citrobacter koseri/farmeri. Isolates presented concomitant resistance to nalidixic acid, ciprofloxacin, ofloxacin, and pefloxacin. A diversity in mutation patterns in gyrA and parC genes and new amino-acid substitutions in GyrA subunit were observed. Quinolone plasmidic resistance genes qnrB1, qnrB2, qnrB5/19, qnrS1, and aac(6')-Ib-cr were detected. Resistance to other antibiotic classes was observed. Class 1 integrons and resistance genes blaCTX-M-15, blaOXA-1, sul1, sul2, sul3, tetA, tetB, aadA1/2, aadA5, aph(3')-Ia, aac(3)II, dfrA1, dfrA5, dfrA7, and dfrA12 were detected. Bacterial tolerance to cadmium, zinc, and mercury was observed with the presence of the merA gene. E. coli isolates belonged to phylogenetic groups A, B1, and D and to sequence types ST405, ST443, ST101, ST10, and ST347. CONCLUSIONS: This study highlighted bacterial multidrug resistance linked to ciprofloxacin and, consequently, the risk of bacterial exposure to this antibiotic.

Characterization of staphylococci in urban wastewater treatment plants in Spain, with detection of methicillin resistant Staphylococcus aureus ST398.

The objective of this study was to determine the prevalence of Staphylococcus in urban wastewater treatment plants (UWTP) of La Rioja (Spain), and to characterize de obtained isolates. 16 wastewater samples (8 influent, 8 effluent) of six UWTPs were seeded on mannitol-salt-agar and oxacillin-resistance-screening-agar-base for staphylococci and methicillin-resistant Staphylococcus aureus recovery. Antimicrobial susceptibility profile was determined for 16 antibiotics and the presence of 35 antimicrobial resistance genes and 14 virulence genes by PCR. S. aureus was typed by spa, agr, and multilocus-sequence-typing, and the presence of immune-evasion-genes cluster was analyzed. Staphylococcus spp. were detected in 13 of 16 tested wastewater samples (81%), although the number of CFU/mL decreased after treatment. 40 staphylococci were recovered (1-5/sample), and 8 of them were identified as S. aureus being typed as (number of strains): spa-t011/agr-II/ST398 (1), spa-t002/agr-II/ST5 (2), spa-t3262/agr-II/ST5 (1), spa-t605/agr-II/ST126 (3), and spa-t878/agr-III/ST2849 (1). S. aureus ST398 strain was methicillin-resistant and showed a multidrug resistance phenotype. Virulence genes tst, etd, sea, sec, seg, sei, sem, sen, seo, and seu, were detected among S. aureus and only ST5 strains showed genes of immune evasion cluster. Thirty-two coagulase-negative Staphylococcus of 12 different species were recovered (number of strains): Staphylococcus equorum (7), Staphylococcus vitulinus (4), Staphylococcus lentus (4), Staphylococcus sciuri (4), Staphylococcus fleurettii (2), Staphylococcus haemolyticus (2), Staphylococcus hominis (2), Staphylococcus saprophyticus (2), Staphylococcus succinus (2), Staphylococcus capitis (1), Staphylococcus cohnii (1), and Staphylococcus epidermidis (1). Five presented a multidrug resistance phenotype. The following resistance and virulence genes were found: mecA, lnu(A), vga(A), tet(K), erm(C), msr(A)/(B), mph(C), tst, and sem. We found that Staphylococcus spp. are normal contaminants of urban wastewater, including different lineages of S. aureus and a high diversity of coagulase-negative species. The presence of multiple resistance and virulence genes, including mecA, in staphylococci of wastewater can be a concern for the public health.

Detection of airborne allergen (Pla a 1) in relation to Platanus pollen in Córdoba, South Spain.

Córdoba is one of the Spanish cities with the highest records of plane tree pollen grains in the air. Clinical studies have identified Platanus as a major cause of pollinosis. This fact provokes an important public health problem during early spring when these trees bloom. The objective of the study is to evaluate the correlation between airborne pollen counts and Pla a 1 aeroallergen concentrations in Córdoba, to elucidate if airborne pollen can be an accurate measure that helps to explain the prevalence of allergenic symptoms. Pollen sampling was performed during 2011-2012 using a Hirst-type sampler. Daily average concentration of pollen grains (pollen grains/m 3 ) was obtained following the methodology proposed by the Spanish Aerobiology Network. A multi-vial cyclone was used for the aeroallergen quantification. Allergenic particles were measured by ELISA using specific antibodies Pla a 1. The trend of Platanus pollen was characterized by a marked seasonality, reaching high concentrations in a short period of time. Airborne pollen and aeroallergen follow similar trends. The overlapping profile between both variables during both years shows that pollen and Pla a 1 are significantly correlated. The highest significant correlation coefficients were obtained during 2011 and for the post peak. Although some studies have found notable divergence between pollen and allergen concentrations in the air, in the case of Platanus in Córdoba, similar aerobiological dynamics between pollen and Pla a 1 have been found. Allergenic activity was found only during the plane tree pollen season, showing a close relationship with daily pollen concentrations. The obtained pollen potency was similar for both years of study. The results suggest that the allergenic response in sensitive patients to plane tree pollen coincide with the presence and magnitude of airborne pollen.

Enterococcus faecium resistente a glucopéptidos. Análisis del genotipode resistencia, virulencia y líneas genéticas / Glycopeptide-resistant Enterococcus faecium. Analysis of the resistance genotype, virulence and genetic lines

Objetivo: El objetivo de este estudio fue analizar las características genotípicas de los aislados de Enterococcus con resistencia adquirida a vancomicina (ERV) obtenidos durante un período de 3 años y 2 meses en el Hospital Clínic de Barcelona. Métodos: Se incluyeron en el estudio todos los aislados de ERV obtenidos en el período de enero de2004 a marzo de 2007. Se analizó el mecanismo de resistencia a vancomicina y las resistencias asociadas a otros antibióticos. Los aislados ERV fueron tipificados por electroforesis en campo pulsante (PFGE) ymulti-locus-sequence-typing (MLST).Resultados: Se obtuvieron 39 ERV que fueron identificados como Enterococcus faecium y representaron el 2% del total de enterococos aislados durante el período de tiempo estudiado. El genotipo van A fue detectado en 38 de los aislados y el genotipo vanB2 en uno adicional. Los 39 ERV fueron clasificados en 13 pulsotipos diferentes (A-M) por PFGE, incluyendo un pulsotipo principal, A, que agrupaba 13 aislados. La secuencia tipo fue identificada por MLST en 24 de las cepas (con patrones diferentes o estrechamente relacionados) y todas ellas fueron adscritas al complejo clonal CC17, excepto 2 que fueron adscritasal complejo CC9. Todas las cepas mostraron un fenotipo de multirresistencia, incluyendo en muchos casos ampicilina, ciprofoxacina, eritromicina, estreptomicina, gentamicina, kanamicina y cloranfenicol,albergando múltiples genes de resistencia asociados. Los genes esp y/o hyl fueron detectados en 37 de losERV. Conclusión: Todas las cepas, excepto una, presentaron el genotipo van A y formaban parte mayoritariamente del complejo clonal CC17 (AU)

Objective: The objective of this study was to analyse the genotypic characteristics of all Enterococcusisolates with acquired vancomycin resistance (VRE) recovered in the Hospital Clinic (Barcelona, Spain)in a period of three years and two months. Methods: All VRE isolated in the referred Hospital in the period January 2004-March 2007 were included in the study. The vancomycin resistance mechanism was investigated, as well as other antibiotic resistance mechanisms. Isolates were also typed by pulsed-fleld-gel-electrophoresis (PFGE) and multilocus-sequence-typing (MLST).Results: Thirty-nine VRE were recovered, all being identifled as E. faecium, representing 2% of total enterococci obtained in that period. Thirty-eight of them carried the van A gene, and one isolate the vanB2 gene.T he 39 VRE were classifled into 13 different pulsotypes (A-M), with one main pulsotype, A, which included 13 isolates. The sequence type was identifled by MLST in 24 VRE (with unrelated or closely-related PFGE patterns), and they were ascribed to the clonal complex CC17, but two classifled as CC9. All VRE showed a multiresistance phenotype, including, in most cases ampicillin, ciprofloxacin, erythromycin, streptomycin, gentamicin, kanamycin and chloramphenicol, harbouring multiple antibiotic resistance genes. The presence of esp and/or hyl genes was identifled in 37 VRE. Conclusion: All VRE, but one, showed the van A genotype and they were mostly ascribed to the high-riskclonal complex CC17 (AU)

Dynamic of nasal colonization by methicillin-resistant Staphylococcus aureus ST398 and ST1 after mupirocin treatment in a family in close contact with pigs.

Nasal colonization by methicillin-resistant Staphylococcus aureus (MRSA) was evaluated after a mupirocin treatment in a family previously colonized by MRSA sequence type ST398 and ST1, who lived close to a pig farm. Eight nasal samples were swabbed from each of the four family members on different moments after mupirocin treatment. The efficacy of treatment was low in those family members who worked in the farm, and higher in the remaining two family members with sporadic contact with pigs. In addition, nasal and skin swabs from randomly selected pigs of the farm were taken. MRSA were detected in 33% of pigs tested. All MRSA isolates obtained were characterized by Staphylococcal-Cassette-Chromosome mec (SCCmec) determination, Multilocus-Sequence-Typing (MLST), spa- and agr-typing, Pulsed-field-gel-electrophoresis (PFGE), antimicrobial susceptibility, detection of antimicrobial resistance genes, and toxin gene profiling. Spa-types t011, t1255 and t1197 were detected in humans and animals, with indistinguishable PFGE patterns, suggesting animal to human MRSA transmission. Each spa-type was ascribed to a specific pulsotype. Spa-types t127 and t108 were only detected in MRSA isolates obtained from humans, and t012 only in those from animals. MRSA ST1-t127 isolates and some ST398-t011 and ST398-t1197 isolates presented a multiantimicrobial-resistance phenotype. None of them harbored lukF/lukS, tst, eta and etb virulence genes. This study showed that the efficacy of nasal MRSA decolonization in healthy people with very close contact with pigs is especially low.

Vancomycin-resistant enterococci from Portuguese wastewater treatment plants.

The objective of this study was to evaluate the incidence of vancomycin resistant enterococci in sludge and sewage of urban and poultry-slaughterhouse wastewater treatment plants. A total of 17 vancomycin resistant enterococci (eight vanA -containing Enterococcus faecium and nine vanC1/vanC2 -containing Enterococcus gallinarum/casseliflavus) were found among 499 isolates of sewage and sludge samples of 14 urban and nine poultry-slaughterhouse wastewater treatment plants. These seventeen VRE isolates showed resistance to kanamycin (n = 8), tetracycline (n = 7), erythromycin (n = 7), ciprofloxacin (n = 7), ampicillin (n = 7), streptomycin (n = 6), and gentamicin (n = 2). The tetM gene, related with tetracycline resistance, was found in six of eight van A-containing isolates, in all seven vanC-1 isolates and in one of two vanC-2 isolates. The ermB gene in seven erythromycin-resistant isolates; and the aac6 '-aph2 ″ gene in the two high-level-gentamicin-resistant isolates. Moreover, two vanA -containing E. faecium isolates harbored the hyl virulence gene, and three isolates the entA bacteriocin gene. The purK-1 allele was detected in our urban vanA -containing E. faecium isolate, and we found as well the purK-6 allele in one poultry-slaughterhouse vanA -containing E. faecium isolate. This study suggests that the wastewater treatment plants might be an important source of dissemination of antibiotic-resistant enterococci in Portugal.

In vivo selection of aac(6')-Ib-cr and mutations in the gyrA gene in a clinical qnrS1-positive Salmonella enterica serovar Typhimurium DT104B strain recovered after fluoroquinolone treatment.

OBJECTIVES: To characterize the mechanisms implicated in the in vivo selection of quinolone and aminoglycoside resistance in a faecal Salmonella enterica serovar Typhimurium DT104B strain recovered after ciprofloxacin treatment of a hospitalized elderly patient with acute gastroenteritis. METHODS: Two Salmonella Typhimurium isolates were obtained before (Se6) and after (Se20) treatment and they were typed by PFGE and multilocus sequence typing (MLST). Antimicrobial susceptibility was determined by disc diffusion and agar dilution methods. Class 1, 2 and 3 integrons and resistance mechanisms were studied by PCR and sequencing. Plasmids were typed. RESULTS: Both Salmonella Typhimurium strains were resistant to tetracycline, streptomycin and sulphonamides, while Se20 was also resistant to nalidixic acid, ciprofloxacin, norfloxacin, levofloxacin, ofloxacin, amikacin, tobramycin, kanamycin and trimethoprim. PFGE and MLST showed a clonal relationship between the strains, which belonged to the sequence type ST36. Both strains contained the repC-sul2-strA-strB structure and tet(A) and qnrS1 genes, and strain Se20 also contained the aac(6')-Ib-cr gene, the Ser83-->Tyr substitution in GyrA and one class 1 integron with the dfrA17 + aadA5 gene cassette arrangement lacking qacEDelta1 + sul1. Two different transconjugants from Salmonella Se20 (TCSe20B and TCSe20L) harboured qnrS1 and sul2 genes and the class 1 integron. The TCSe20B strain also acquired the aac(6')-Ib-cr gene located on a non-typeable plasmid. qnrS1 was identified on a ColE-type plasmid and the class 1 integron on an IncI1-type plasmid. CONCLUSIONS: This is the first report of in vivo selection of the aac(6')-Ib-cr gene and the Ser83-->Tyr change in GyrA in a qnrS1-positive Salmonella Typhimurium strain after ciprofloxacin treatment; the in vitro transfer of both plasmid-mediated quinolone resistance genes was also demonstrated.

Axenfeld-Rieger ocular anomaly and retinoblastoma caused by constitutional chromosome 13q deletion.

Axenfeld-Rieger (AR) ocular anomaly might be due to deletions of different chromosomes. No association between AR, mental retardation, and retinoblastoma has been described. We report a 2-month-old female with general development delay and dysmorphic features. AR anomaly was detected, and a retinoblastoma (RB) was diagnosed in a very early stage. De novo 13q deletion was identified. Systemic chemotherapy, focal cryotherapy, transpupillary thermotherapy, brachytherapy, and intra-arterial chemotherapy were needed to control the RB. This is the first report of an association of AR, 13q deletion, and retinoblastoma, to be disclosed in patients born with such ocular and dysmorphic features.