Antimicrobial resistance profiles of Enterococcus faecalis and Enterococcus faecium isolated from artisanal food of animal origin in Argentina.

Enterococci are part of the indigenous microbiota of human gastrointestinal tract and food of animal origin. Enterococci inhabiting non-human reservoirs play a critical role in the acquisition and dissemination of antimicrobial resistance determinants. The aim of this work was to investigate the antimicrobial resistance in Enterococcus faecalis and Enterococcus faecium strains recovered from artisanal food of animal origin. Samples of goat cheese (n = 42), cow cheese (n = 40), artisanal salami (n = 30), and minced meat for the manufacture of hamburgers (n = 60) were analyzed. Phenotypic and genotypic tests for species-level identification of the recovered isolates were carried out. Minimum inhibitory concentration (MIC) study for in vitro quantitative antimicrobial resistance assessment was performed, and 71 E. faecalis and 22 E. faecium were isolated. The recovered enterococci showed different multi-drug resistance patterns that included tretracycline, erythromycin, ciprofloxacin, linezolid, penicillin, ampicillin, vancomycin, teicoplanin, gentamicin (high-level resistance), and streptomycin (high-level resistance). VanA-type E. faecium were detected. ß-lactamase activity was not observed. Artisanal foods of animal origin act as a non-human reservoir of E. faecalis and E. faecuim strains, expressing multi-resistance to antimicrobials. In conclusion, the implementation of a continuous antimicrobial resistance surveillance in enterococci isolated from artisanal food of animal origin is important.

Detection of ß-Lactamase-Producing Enterococcus faecalis and Vancomycin-Resistant Enterococcus faecium Isolates in Human Invasive Infections in the Public Hospital of Tandil, Argentina.

The study's aim was to analyze the population structure of enterococci causing human invasive infections in a medium-sized Argentinian Hospital coincidental with a 5 year-period of increased recovery of antibiotic resistant enterococci (2010-2014). Species identification (biochemical testing/MALDI-TOF-MS), antimicrobial susceptibility (disk-diffusion) and clonal relatedness (PFGE/MLST/BAPS) were determined according to standard guidelines. ß-lactamase production was determined by a nitrocefin test and confirmed by PCR/sequencing. The isolates were identified as Enterococcus faecalis and Enterococcus faecium at a 2:1 ratio. Most of the E. faecalis isolates, grouped in 25 PFGE-types (ST9/ST179/ST236/ST281/ST388/ST604/ST720), were resistant to high-levels (HLR) of gentamicin/streptomycin. A ST9 clone (bla+/HLR-gentamicin) was detected in patients of different wards during 2014. E. faecium isolates were grouped in 10 PFGE-types (ST25/ST18/ST19/ST52/ST792), with a low rate of ampicillin resistance. Five vancomycin-resistant E. faecium, three vanA (ST792/ST25) and two vanB (ST25) were detected. The ST25 clone carried either vanA or vanB. The recovery of a bla+-ST9-E. faecalis clone similar to that described in the late 1980s in Argentina suggests the possibility of a local hidden reservoir. These results reflect the relevance of local epidemiology in understanding the population structure of enterococci as well as the emergence and spread of antimicrobial resistance in predominant enterococcal clonal lineages.

Comparative time-kill study of doxycycline, tigecycline, sulbactam, and imipenem against several clones of Acinetobacter baumannii.

To assess potential alternative options for the treatment of infections caused by Acinetobacter baumannii, we performed time-kill studies of doxycycline and tigecycline using several isolates recovered from patients residing in 10 different cities in Argentina. Imipenem and sulbactam were also included for comparison purposes. Eleven isolates representing 5 distinctive clones, or isolates with different susceptibility patterns within the same clone, were selected. Tubes containing cation-supplemented Mueller-Hinton broth with and without antibiotics were seeded with a log-phase inoculum of roughly 5 x 10(5) CFU/mL. By using the viable counts determined at 2-, 4-, 6-, 8-, and 24-h intervals after inoculation, a 24-h time-kill curve was constructed for each isolate. No bactericidal activity (defined as a >or=3-log(10) CFU/mL decrease in the viable cell counts with respect to the original inoculum) was observed at any time with sulbactam (4 microg/mL) or tigecycline (1 microg/mL), whereas low bactericidal rate (18% of the isolates) was shown for doxycycline (1 microg/mL) and sulbactam (16 microg/mL) after 24 h of incubation. Doxycycline (4 microg/mL) and tigecycline (8 microg/mL) displayed bactericidal activity at 24 h of incubation against 36% and 54% of the isolates, respectively, including the carbapenem-resistant isolate. Corresponding values for imipenem (1 and 4 microg/mL) against the 10 carbapenem-susceptible isolates were 60% and 90%, respectively. The present study confirms the in vitro efficacy of imipenem against A. baumannii, suggests that doxycycline could be a suitable, cost-effective, alternative option in some instances, and sheds light on the potential role of tigecycline in the treatment of infections with this organism.

Características clínicas y microbiológicas de infecciones de piel y partes blandas en pacientes pediátricos de dos hospitales de Buenos Aires / Skin and soft tissue infections in pediatrics

Las infecciones de piel y partes blandas (IPPB) en niños son una de las principales causas de prescripción de antimicrobianos. El objetivo del estudio fue describir las características clínicas y microbiológicas de las IPPB ambulatorias de niños asistidos en dos hospitales zonales. Se realizó un estudio prospectivo entre el 1/11/2017 y el 1/11/2018. Se incluyeron pacientes entre 1 mes y 15 años internados en dos hospitales. Se evaluó: edad, sexo, localidad, factores predisponentes, tipo de IPPB, muestras biológicas realizadas, aislamiento microbiológico, tratamiento empírico indicado y evolución del cuadro. Se realizó antibiograma y determinación genética. Se calculó chi2, IC95, OR; α=5%. N= 94. 58,7% masculinos. 12 pacientes <1 año, 85 >1 año (promedio de edad 4 años, 1-15). El 36% de Tandil y 63,8% de Florencio Varela. El 59,6% corresponden a IPPB purulentas. Se aislaron microorganismos en un 59,6%. Los aislamientos principales: SAMR (40,4%), SAMS (7,4%), S. agalactiae (2,1%) y S. pyogenes (2,1%). El 100% de SAMR son portadores de gen mecA y SCCmec tipo IV, sin multirresistencia. No hubo diferencia estadística entre los factores de riesgo evaluados para el desarrollo de IPPB por SAMR. El 52,1% de los niños recibió tratamiento antibiótico combinado, siendo la más indicada TMS-SMX + CLI en 36 eventos. (38,3%). La evolución fue favorable: no hubo diferencia significativa entre el subgrupo que se aisló SAMR y el que no se aisló SAMR; 91,9% (34/37) y 92,6% (50/54) correspondientemente (chi2: 0,01; p= 0,97 IC95: 0,26-3,88). El principal agente etiológico fue SAMRco, debiendo adecuar los tratamientos a este microorganismo.

Skin and soft tissue infections (SSIs) in children are one of the main causes of antimicrobial prescription. The aim of the study was to describe the clinical and microbiological characteristics of outpatient SSIs in children attended in two hospitals. A prospective study was conducted between 11/1/2017 and 11/1/2018. Patients between 1 month and 15 years old, hospitalized were included. We evaluated: age, sex, locality, predisposing factors, type of IPPB, biological samples taken, microbiological isolation, empirical treatment indicated and evolution of the condition. An antibiogram and genetic determination were performed. Chi2, CI95, OR; α=5% were calculated. N= 94. 58.7% male. 12 patients <1 year, 85 >1 year (mean age 4 years, 1-15). 36% were from Tandil and 63.8% from Florencio Varela. 59.6% corresponded to purulent SSIs. The diagnostic yield was 59.6%. Main isolates: MRSA (40.4%), MSSA (7.4%), S. agalactiae (2.1%) and S. pyogenes (2.1%). 100% of MRSA carried the mecA gene and SCCmec type IV, with no multidrug resistance. There was no statistical difference between the risk factors evaluated. 52.1% of children received combined antibiotic treatment, the most indicated being TMS-SMX + CLI in 36 events. (38,3%). Evolution was favorable: there was no significant difference between the subgroup that isolated MRSA and the subgroup that did not isolate MRSA; 91.9% (34/37) and 92.6% (50/54) respectively (chi2: 0.01; p= 0.97 CI95: 0.26-3.88). The main etiological agent was MRSA, and treatments should be adapted to this microorganism

Bacteriemia por Staphylococcus aureus con sensibilidad disminuida heterogénea a vancomicina: reporte de un caso / Staphylococcus aureus bacteremia with heterogeneous intermediate-level vancomycin resistance: a case report

Staphylococcus aureusmeticilina-resistente (SAMR)es una causa frecuente de bacteriemias intrahospitalarias. Para su tratamiento se utiliza vancomicina y han emergido cepas con sensibilidad disminuida heterogénea (h-VISA) que albergan subpoblaciones con sensibi-lidad reducida a vancomicina. Se comunica un caso de bacteriemia intra-tratamiento con vancomicina por SAMR h-VISA. El aislamiento muestra sensibilidad a vancomicina (CIMvan: 1 µg/mL), sin embargo E-test GRD sugiere h-VISA (CIMvan: 2 µg/mL y CIMtei: 8 µg/mL). El análisis del perfil poblacional - área bajo la curva (PAP-AUC) valida este hallazgo. Se rota a linezolid con resolución clínica.

Methicillin-resistant Staphylococcus aureus (SAMR) is a common cause of nosocomial bacteremia. Vancomycin, a glycopeptide, is widely employed for the therapy of SAMR infections. In recent years, heterogeneous vancomycin-intermediate strains (h-VISA) have emerged. We report a case of intra-treatment bacteremia caused by SAMR h-VISA. The isolate shows susceptibility to vancomycin (MICvan: 1 µg/mL). But the GRD E-test suggests h-VISA (MICvan: 2 µg/mL and MICtei: 8 µg/mL). The population analysis profile - area under the curve (PAP-AUC) validates SAMR h-VISA. Rotation of antibiotic therapy with linezolid is done, with good clinical outcome.

Caracterización fenotípica de variantes pequeñas de Staphylococcus aureus: un nuevo desafío / Phenotypic characterization of small colony variants of Staphylococcus aureus: a new challenge / Caracterizaçao fenotípica de pequenas variantes de Staphylococcus aureus: um novo desafio

En infecciones crónicas y recurrentes por Staphylococcus aureus se han descripto subpoblaciones de colonias pequeñas (VCPSa). El objetivo de este trabajo fue reconocer las características fenotípicas de VCPSa para optimizar su detección y caracterización a partir de materiales clínicos provenientes de infecciones crónicas. Se analizaron n=3 VCPSa de pacientes adultos con infecciones crónicas de tejidos blandos. Las muestras se inocularon en agar nutritivo, agar sangre, agar chocolate y agar Schaedler suplementado. Se realizaron tinción de Gram, catalasa, coagulasa libre, pruebas de dependencia para hemina, menadiona y timidina y, desarrollo/ataque del manitol en agar manitol salado. La sensibilidad antibiótica se efectuó en agar Mueller Hinton suplementado, según las pruebas de dependencia. Se investigó la presencia de proteína ligadora de penicilina anómala (PBP2´) por aglutinación con látex. Las VCPSa se detectaron en los medios de cultivo enriquecidos. Estas bacterias dieron positivas las pruebas de catalasa y coagulasa, y eran dependientes de menadiona y hemina. En los tres aislamientos se observó resistencia a cefoxitina y se detectó la PBP2´.

In chronic and recurrent infections, small colonies of Staphylococcus aureus subpopulations (SCVSa) have been observed. The objective of the present study was to recognize the phenotypic characteristics of SCVSa isolated from patients with chronic infections to optimize their detection. SCVSa of adult patients n=3 with chronic soft tissue infections were analyzed. Samples were inoculated on nutritive agar, blood-agar, chocolate agar and supplemented Schaedler agar. Subsequently, Gram stain, catalase, free coagulase, dependence tests for hemin, menadione and thymidine, and growth/fermentation of mannitol on salt mannitol agar were performed. Antibiotic susceptibility tests were performed by the agar diffusion method on supplemented Mueller Hinton agar, according to dependence assays results. Anomalous penicillin binding protein (PBP2') was investigated by latex agglutination. SCVSa were detected in all enriched culture media. They showed catalase and coagulase activities, and menadione and hemin dependence. By the agar diffusion test, cefoxitin resistance was found in all isolates; PBP2' was detected as well.

Nas infecções crônicas e recorrentes por Staphylococcus aureus, subpopulações de pequenas colônias (VCPSa) foram descritas. O objetivo desse trabalho foi reconhecer as características fenotípicas de VCPSa para otimizar sua detecção e caracterização a partir de materiais clínicos provenientes de infecções crônicas. Foram analisados n=3 VCPSa de pacientes adultos com infecções crônicas de tecidos moles. As amostras foram inoculadas em agar nutritivo, agar sangue; agar chocolate e agar Schaedler enriquecido. Foram realizados testes de coloração de Gram, catalase, coagulase livre, testes de dependência para hemina, menadiona e timidina, e desenvolvimento/fermentação do manitol em agar manitol salgado. A sensibilidade antibiótica foi realizada em agar Mueller Hinton suplementado, de acordo com os testes de dependência. Foi investigada a presença de proteína ligante de penicilina anômala (PBP2´) por aglutinação com látex. Os VCPSa foram detectados em meios de cultura enriquecidos. Estas bactérias deram positivas nos testes de catalase e coagulase positivos e eram dependentes de menadiona e hemina. A resistência à cefoxitina foi detectada nos três isolados e detectou-se a PBP2'.

Enterococcus spp. aislados de queso de oveja: resistencia a los antimicrobianos de utilización clínica / Enterococcus spp. isolated from goat cheese: resistance to clinically used antimicrobials

En Enterococcus spp. aislados de quesos de oveja se investigó la resistencia in vitro a antimicrobianos de utilización clínica. Se determinó la concentración inhibitoria mínima (CIM) para gentamicina, estreptomicina, vancomicina, teicoplanina, ampicilina, imipenem, linezolid y tigeciclina. Se detectó la producción de β-lactamasa y se realizaron experimentos de conjugación para transferir resistencia a gentamicina. En un aislamiento de E. faecalis y en 10 de E. faecium se observó resistencia a ampicilina; se demostró la producción de β-lactamasa en un E. faecalis. En E. faecium, 15 aislamientos fueron resistentes a imipenem; 4 a linezolid y 5 a glucopéptidos. En 4 aislamientos de E. faecalis se demostró alto nivel de resistencia plasmídica a gentamicina. No se observaron aislamientos resistentes a tigeciclina. El queso de oveja es un reservorio de enterococos resistentes con potencial diseminación al hombre a través de la cadena alimentaria.

In vitro resistance to clinically used antimicrobials was investigated in Enterococcus spp. isolated from goat cheese. The gentamycin, streptomycin, vancomycin, teicoplanin, ampicillin, imipenem, linezolid and tigecyclin minimum inhibitory concentration (MIC) was determined. Β-lactamase production was detected and conjugation experiments were carried out for transferring gentamycin resistance. In one E. faecalis and in 10 E. faecium isolates there was ampicillin resistance and β-lactamase production was demonstrated in one E. faecalis. Fifteen E. faecalis were imipenenem resistant; 4 linezolid resistant and 5 glucopeptide resistant. There was a high level of plasmidic resistance to tigecyclin in 4 E. faecalis isolates. There were no tigecyclin resistant isolates. Goat cheese is a reservoir of resistant enterococci with potencial human dissemination through the food chain.

Primer informe de celulitis cutánea por Erysipelothrix rhusiopathiae en dos trabajadores rurales / First report of cutaneous cellulitis caused by Erysipelothrix rhusiopathiae in two rural workers

Se informan dos casos de celulitis cutánea en trabajadores de un mismo criadero de aves a corral del área rural de la ciudad de Tandil, Argentina. En ambos pacientes fue identificado E. rhusiopathiae a partir de cultivos de muestras obtenidas por biopsia. La investigación del reservorio evidenció que en todas las muestras de las excretas de aves y en dos muestras de los residuos del alimento, se aisló E. rhusiopathiae con idéntico perfil bioquímico y de resistencia antimicrobiana al de las cepas aisladas de los dos casos clínicos. Si bien son necesarios estudios moleculares para establecer la relación epidemiológica entre las cepas aisladas de los pacientes y las del criadero, se puede inferir que la fuente de infección humana fue el criadero extensivo de aves de corral.

This report describes two cases of cutaneous cellulitis in workers at the same poultry farm in the rural area of Tandil, Argentina. In both patients E. rhusiopathiae was identified from culture samples obtained through biopsy. Investigation of the reservoir showed that, in all samples of fowl waste and in 2 samples of feed, E. rhusiopathiae was isolated with identical biochemical and antibiotic resistance profiles to those of the isolated strains in the case reports. Even though molecular analyses are required to establish an epidemiologic relation between the strains found in the patients and those on the farm, it can be inferred that the source of human infection was the extensive poultry farm.