[Anaerobiosis beyond anaerobic bacteria: its role in the recovery of aerobic microorganisms from purulent samples]. / La anaerobiosis más allá de las bacterias anaerobias: su importancia en la recuperación de microorganismos aerobios a partir de materiales purulentos.

The main objective of incubation in anaerobiosis is the recovery of obligate anaerobic bacteria, not excluding other microorganisms. In 2003, we conducted a comparative and prospective study from consecutive clinical samples on the recovery of aerobic microorganisms from primary cultures both in anaerobiosis and aerobiosis of the same sample. The aims were to evaluate the methodology used in anaerobiosis in the recovery of aerobic microorganisms not diagnosed in primary aerobic cultures, and to establish a relationship between them and the origin of the sample. From 2003 to 2004, 2776 bacteriological samples were analyzed and 1884 aerobic microorganisms were cultured altogether. The result was that 69.4% of the samples showed growth both in aerobic and anaerobic incubation from primary cultures of the sample, whereas 30.6% only in one of the mentioned incubation atmosphere: 49.2% in aerobiosis and 50.8% in anaerobiosis. According to these results, the methodology used in anaerobiosis (anaerobic incubation, culture media, stereoscopic microscope or hand lens to examine the primary plates), allowed an extra yield of aerobic organisms, especially gram positive facultative and microaerophilic cocci, which was particularly evident in polimicrobial cultures, and especially when gram negative accompanying flora was present, independently of the type of sample.

[Linezolid for the treatment of newborns infected with vancomycin-resistant Enterococcus faecium]. / Uso de linezolid para el tratamiento de recién nacidos con infecciones por Enterococcus faecium resistentes a la vancomicina.

Five patients hospitalized in the neonatal unit between 2002 and 2007, infected with vancomycin-resistant Enterococcus faecium treated with linezolid are presented in this study. This antibiotic showed good clinical efficacy and safety, since no adverse events occured in this group of patients.

Endocarditis due to vancomycin-resistant Enterococcus raffinosus successfully treated with linezolid: case report and review of literature.

Enterococcus raffinosus is scarcely found in clinical samples and even less frequently as etiologic agent of endocarditis. We are herein presenting one case of mitral prosthetic-valve endocarditis in a 77-y-o male due to a vancomycin-resistant Enterococcus raffinosus isolate, successfully treated with 6 weeks of linezolid, and a two-year follow up.

[Bacteremia due to Abiotrophia defectiva in a febrile neutropenic pediatric patient]. / Bacteriemia por Abiotrophia defectiva en un paciente pediótrico neutropénico febril.

The presence of Granulicatella spp. in bacteremic episodes of neutropenic patients was recently highlighted whereas Abiotrophia defectiva, was only isolated in cases of infectious endocarditis. The aim of this study is to describe a case of A.defectiva bacteremia in a leukemic and febrile (40 degrees C) neutropenic (200 GB/mm3) boy. A.defectiva was only isolated from one of the two processed blood samples. Although the patient was undergoing an episode of varicela which could have accounted as the possible cause of fever, A. defectiva was considered a significant finding because this species is not part of the commensal skin flora. This case suggests that both A. defectiva and Granulicatella spp. should be regarded as possible causes of bacteremia in immunocompromised patients.

[Influence of penicillin minimum inhibitory concentration in the synergy between penicillin and gentamicin in viridans-group streptococci]. / Influencia de la concentración inhibitoria mínima de penicilina en la acción sinérgica de su combinación con gentamicina frente a estreptococos del grupo viridans.

Penicillin resistance rates higher than 60% have been recorded in viridans group streptococci by some authors during the 90's and recently such resistance was associated with higher levels of mortality in bacteremia. The lowest minimum inhibitory concentration of penicillin for which synergy with aminoglycosides is not yet possible is still unknown. In order to try to dilucidate this puzzle, a study on the susceptibility to penicillin of 28 strains of viridans group streptococci isolated from significant samples in the Hospital de Pediatría "Prof. Dr. Juan P. Garrahan" was carried out. Seven mitis group isolates presenting different susceptibility patterns were selected for performing time-killing curves with penicillin, gentamicin, and penicillin plus gentamicin, using higher and lower penicillin concentrations than their minimal inhibitory concentrations. Synergy was not observed when the penicillin concentration was lower than the minimum inhibitory concentration, at least in these strains with minimum inhibitory concentrations of gentamicin > or = 16 microg/ml. When using penicillin in higher concentrations than the minimum inhibitory concentration, synergy was found in five of the seven strains. Aminoglycoside-modifying enzymes were found in the two other streptococci.

Whole-cell protein profiles are useful for distinguishing enterococcal species recovered from clinical specimens.

Whole-cell protein analysis was performed for differentiating 150 enterococcal isolates to the species level, which had previously been identified by extended phenotypic conventional tests. Whole-cell protein profile (WCPP) showed a high degree of similarity within species and comparison between species revealed important differences in band profiles. All Enterococcus faecalis and Enterococcus faecium isolates were properly located into their corresponding species, regardless of their clinical source and susceptibility pattern. Moreover, WCPP allowed relocation of some isolates that had erroneously been identified by the usual conventional scheme (i.e. two atypical arginine-negative E. faecalis isolates). WCPP proved to be a simple method to ascertain the various enterococcal species, especially those other than E. faecalis, and may be a suitable tool for high-complexity or reference clinical laboratories.

Infecciones por estreptococos del grupo S. anginosus en pediatría / S. anginosus group streptococcal infections in children

Los estreptococos del grupo Streptococcus anginosus (EGA), también llamados "Streptococcus milleri", fueron reconocidos como parte de los estreptococos del grupo viridans (EGV) desde principios del siglo XX. Su rol como patógenos humanos, sin embargo comenzó a destacarse recién en la década de 1970. En esta actualización se describen aspectos microbiológicos y clínicos de los EGA. Los métodos fenotípicos de identificacón e incluso algunos genotípicos carecen de precisión para reconocer las tres especies del grupo (Streptococcus anginosus, Streptococcus constellatus y Streptococcus intermedius) e incluso pueden fallar en su clasificación a nivel de grupo. La mayoría de ellos son sensibles a los antibióticos beta-lactámicos pero son considerables los porcentajes de resistencia a macrólidos, lincosamidas y tetraciclinas. Los EGA son colonizantes habituales de las mucosas orofaríngea, intestinal y genitourinaria, pero, cada vez más frecuentemente, son reconocidos como patógenos humanos. Es ampliamente conocida su capacidad de formar abscesos en órganos sólidos, especialmente abscesos cerebrales, pulmonares y hepáticos. También producen sinusitis, empiemas y colecciones en piel y tejidos blandos, hueso, articulaciones, etc. Se han encontrado asociados con exacerbaciones pulmonares en pacientes con fibrosis quística y con enfermedad pulmonar obstructiva crónica. Producen también infecciones posteriores a mordeduras humanas, infecciones diseminadas, bacteriemia sin foco aparente y, en menor medida, endocarditis infecciosa (AU)

Streptococci from the Streptococcus anginosus group (SAG), also termed "Streptococcus milleri", were recognized as members of the viridans group streptococci (VGS) in the early 20th century. Nevertheless, their role as human pathogens only became evident in the 1970s. In this update, microbiological and clinical aspects of the SAG are described. Phenotypic and even some genotypic identification methods lack accuracy in recognizing the three species of the group (Streptococcus anginosus, Streptococcus constellatus, and Streptococcus intermedius) and may fail to classify them at the group level. Most of them are sensitive to beta-lactam antibiotics but rates of resistance to macrolides, lincosamides, and tetracyclines are significant. SAGs are common colonizers of the oropharyngeal, intestinal, and genitourinary mucosa, but are increasingly recognized as human pathogens. Their ability to form abscesses in solid organs, especially brain, lung and liver, is widely known. They may produce sinusitis, empyemas, and collections in skin and soft tissues, bone, joints, etc. They have also been associated with pulmonary exacerbations in patients with cystic fibrosis and chronic obstructive pulmonary disease. In addition, they may cause infections following human bites, disseminated infections, bacteremia without apparent focus, and, to a lesser extent, infective endocarditis (AU)

Sensibilidad disminuida a los antibóticos Beta-Lactamicos en estreptococos Beta-Hemolíticos / Decreased sensitivity to beta-lactam antibiotics in beta-hemolytic streptococci

Últimamente, se están detectando mutaciones en las proteínas ligadoras de penicilina (PBP) de los estreptococos beta-hemolíticos que corresponden a sitios que en Streptococcus pneumoniae han determinado sensibilidad disminuida a los antibióticos beta-lactámicos. Primero, se describieron cepas con sensibilidad intermedia a penicilina en Streptococcus agalactiae (estreptococos del grupo B), luego en Streptococcus dysgalactiae subsp. equisimilis (mayormente grupos C y G) y, más recientemente, cepas con sensibilidad disminuida a aminopenicilinas y cefalosporinas de tercera generación en Streptococcus pyogenes (grupo A). El costo biológico de estas modificaciones nos permite pensar que los niveles de resistencia no han de ser tan elevados como para comprometer por ahora la efectividad clínica de los beta-lactámicos (AU)

Recently, mutations in penicillin-binding proteins (PBPs) of beta-hemolytic streptococci have been detected corresponding to sites that in Streptococcus pneumoniae have been determined to have decreased sensitivity to beta-lactam antibiotics. First, strains with intermediate sensitivity to penicillin were described in Streptococcus agalactiae (group B streptococci), subsequently in Streptococcus dysgalactiae subsp. equisimilis (mainly groups C and G) and, more recently, strains with decreased sensitivity to third-generation aminopenicillins and cephalosporins were found in Streptococcus pyogenes (group A). The biological cost of these modifications suggests that, for now, resistance levels are not high enough to compromise the clinical effectiveness of beta-lactams (AU)

First report of VanA Enterococcus gallinarum dissemination within an intensive care unit in Argentina.

Enterococcusgallinarum is intrinsically resistant to low levels of vancomycin and has been described as a colonizing microorganism causing bacteraemia and infection among immunosupresed patients. Between August 2000 and February 2001, 15 highly glycopeptide-resistant E. gallinarum isolates, one from blood and the remaining from rectal swabs, were recovered in a general hospital of Buenos Aires Province, Argentina. All isolates were characterized by biochemical assays, and displayed MICs of vancomycin in the range 16-128 mg/l and MICs of teicoplanin in the range 16-32 mg/l. In all cases, PCR analysis yield positive results for both vanC1 and vanA genes. E. gallinarum isolates were classified as two clonal types by SmaI-PFGE: clone A (n = 8) and clone B (n = 7) and both harboured a transferable vanA element.

[Fatal mitral valve endocarditis by Erysipelothrix rhusiopathiae]. / Endocarditis fatal con localizaci'on mitral producida por Erysipelothrix rhusiopathiae.

A fatal case of Erysipelothrix rhusiopathiae mitral valve endocarditis is described in a 45 years old male, with a history of chronic alcohol abuse and without animals contact. He presented intermittent fever, polyarthralgia, weight loss, and low back pain. In blood cultures (2 bottles), gram-positive pleomorphic rods grew after 48 hours of incubation. The subculture on blood agar media showed a small, alpha-hemolytic colony, catalase and oxidase negative, PYR and LAP positive and the production of H2S in triple sugar iron agar, was demonstrated. The isolate was initially identified as E. rhusiopathiae, and confirmed by API Coryne (BioMérieux). On the basis of these findings and a transthoracic echocardiogram, an endocarditis was confirmed. Intravenous ampicillin and gentamicin treatment was initiated. The patient became afever, nevertheless he died on day 19 after admission as a consequence of acute pulmonary edema. Susceptibility testing by E-test showed that the microorganism was resistant to vancomycin and gentamicin, and susceptible to penicillin and cefotaxime. We emphasize the importance to consider the isolates of gram-positive pleomorphic rods, catalase and oxidase negative, and the addition of H2S production test in TSI medium. Vancomycin-resistance helps in the identification, and to establish the correct antimicrobial therapy. Although E. rhusiopathiae is usually reported as an occupational pathogen, the contact with pigs and other farm animals may be underestimated.

[Clinical and epidemiologic analysis of intestinal tract colonization with vancomycin-resistant enterococci in an intensive care unit]. / Análisis clínico-epidemiológico de la portación intestinal de enterococos resistentes a vancomicina en una unidad de terapia intensiva.

Intestinal tract colonization with vancomycin resistant enterococci (VRE) was studied during five months and 25 days. Out of 171 patients hospitalized in the intensive care unit, 124 (73%) were included in this study. Thirty five of them (28%) were recognized as colonized with VRE. VRE isolates (n = 35) were identified as Enterococcus faecium (n = 18), Enterococcus gallinarum (n = 16), and Enterococcus raffinosus (n = 1). All of them were resistant to vancomycin (MIC90 = 512 microg/ml) and to teicoplanin (MIC90 = 32 microg/ml), having the vanA gene. By means of molecular methods a high homology was found among E. faecium and E. gallinarum isolates, respectively, suggesting their spread as a kind of outbreak. No significant differences in age or sex were found among colonized and non-colonized patients (p > 0.05). On the other hand, the hospitalization time and the use of broad-spectrum antibiotics were associated with colonization. From this study we highlight the importance of enhancing all measures of control and prevention of hospital infections, carefully analyzing the empiric antimicrobial schemes, trying to reduce the hospital stage, and following the surveillance to evaluate the efficacy of such procedures.

[Consensus for antimicrobial susceptibility testing for Enterobacteriaceae. Subcommittee on Antimicrobials, SADEBAC (Argentinian Society of Clinical Bacteriology), Argentinian Association of Microbiology]. / Consenso sobre las pruebas de sensibilidad a los antimicrobianos en Enterobacteriaceae.

Taking into account previous recommendations from the National Committee for Clinical Laboratory Standards (NCCLS), the Antimicrobial Committee, Sociedad Argentina de Bacteriología Clínica (SADEBAC), Asociación Argentina de Microbiología (AAM), and the experience from its members and some invited microbiologists, a consensus was obtained for antimicrobial susceptibility testing and interpretation in most frequent enterobacterial species isolated from clinical samples in our region. This document describes the natural antimicrobial resistance of some Enterobacteriaceae family members, including the resistance profiles due to their own chromosomal encoded beta-lactamases. A list of the antimicrobial agents that should be tested, their position on the agar plates, in order to detect the most frequent antimicrobial resistance mechanisms, and considerations on which antimicrobial agents should be reported regarding to the infection site and patient characteristics are included. Also, a description on appropriate phenotypic screening and confirmatory test for detection of prevalent extended spectrum beta-lactamases in our region are presented. Finally, a summary on frequent antimicrobial susceptibility profiles and their probably associated resistance mechanisms, and some infrequent antimicrobial resistance profiles that deserve confirmation are outlined.

Viridans streptococci bacteraemia in children with fever and neutropenia: a case-control study of predisposing factors.

Viridans streptococci (VS) are an increasing cause of bacteraemia in neutropenic patients with cancer. Case-control studies of predisposing factors for acquisition of this infection in children are not published. Between January 1989 and December 1999, 168 episodes of bacteraemia in 161 children with fever and neutropenia of haemato-oncology origin were analysed. 15 cases (9%) in 15 patients were caused by VS. Each case patient was compared with 6 matched control patients; 2 with other Gram-positive cocci (group 2), 2 with gram-negative bacilli bacteraemia (group 3) and two children with fever and neutropenia without bacteraemia (group 4). The median age of patients was 4.1 years (range: 2-15 years). 87% of children had acute leukaemia or lymphomas. Pneumonia was the predominant clinical focus (70%). Shock was observed in 13% of patients. ARDS was observed in one child who died of this complication. Multivariate analysis of risk factors for the development of VS bacteraemia showed that two factors were independent predictors: high doses of cytosine-arabinoside (ARA-C) as part of the chemotherapy treatment (Odds Ratio (OR): 9.3; Confidence Interval (CI) 1.56-55.5) (P<0.014) and the presence of pneumonia (OR: 1.36: CI 2.27-81.9) (P<0.0043). We propose that further studies are warranted to confirm these results.

Study to determine the ability of clinical laboratories to detect antimicrobial-resistant Enterococcus spp. in Buenos Aires, Argentina.

Few reports of vancomycin-resistant enterococci have appeared outside the USA. Therefore, we evaluated the ability of five laboratories in Buenos Aires, Argentina, to perform susceptibility testing using the disk diffusion method. Laboratories had difficulty identifying the low- and intermediate-level vancomycin-resistant phenotypes. This suggests that the disk diffusion method used by laboratories abroad may fail to detect some vancomycin-resistant enterococci.

Argentinian collaborative study on prevalence of erythromycin and penicillin susceptibility in Streptococcus pyogenes. The Argentinian Streptococcus Study Group.

Two monthly studies on the prevalence of penicillin and erythromycin susceptibility of Streptococcus pyogenes were performed in May and October of 1994 in Argentina. A total of 58 centers from 27 cities participated in these studies. A total of 1072 isolates were tested by a diffusion method, although 595 isolates were tested both by the diffusion and an agar dilution method (n = 1767 isolates). No penicillin-resistant streptococci were found in our study (MIC100 = 0.03 microgram/ml). Only four isolates were confirmed as erythromycin resistant S. pyogenes (prevalence 0.14 and 0.28% in May and October 1994, respectively). Resistance in three isolates was due to an inducible mechanism, although in one strain a different phenotype was observed.

Comparative analysis of outcome and clinical features in children with pleural empyema caused by penicillin-nonsusceptible and penicillin-susceptible Streptococcus pneumoniae.

OBJECTIVE: To describe the clinical characteristics of pleural empyema in children caused by Streptococcus pneumoniae nonsusceptible to penicillin and compare their clinical outcome with pleural empyemas caused by penicillin-susceptible organisms. METHODS: Records of 109 children with complicated pleural effusions between January 1996 and December 1998 were retrospectively reviewed. RESULTS: Pathogens were recovered in 45 of these cases. Streptococcus pneumoniae represented 71% of the isolates, with 20 (62%) and 12 (38%) of the strains proving susceptible and nonsusceptible to penicillin, respectively. The average age for children with penicillin-nonsusceptible infections was 18.3 months compared with an average age of 40.9 months for those with penicillin-susceptible infections (P = 0.03). Previous antibiotic treatment was more frequent in children with penicillin-nonsusceptible organisms (P = 0.05). No significant differences were found between the two groups for gender, underlying diseases, duration of fever and tachypnea, need of surgical treatment, bacteremia incidence, mean duration of therapy, or length of hospital stay. CONCLUSIONS: Pleural empyemas caused by penicillin-nonsusceptible S. pneumoniae appear to be associated with younger age and previous antibiotic treatment. By contrast, the outcome was not influenced by the penicillin susceptibility of the pneumococcus strains.

[In vitro activity of trovafloxacin, of other fluoroquinolones and of related antimicrobials against clinical isolates. Grupo colaborativo WHONET-Argentina]. / Actividad in vitro de trovafloxacina, otras fluoroquinolonas y de diferentes antimicrobianos frente a aislamientos clínicos. Grupo colaborativo WHONET-Argentina.

The in vitro activity of trovafloxacin (TRV) has been evaluated in comparison with that of other antimicrobial agents against 5671 clinical isolates recovered by representative institutions of different provinces in our country. The resistance percentage to gentamicin and third generation cephalosporins among enterobacteriaceae was high: 17% and 16% respectively, with a considerable variation according to the analyzed species. The resistance to ciprofloxacin (CIP) and TRV affected approximately 9% of the isolates, without significant differences between both drugs. Fluoroquinolones (FQ) presented excellent activity on 166 isolates of Salmonella spp., 208 of Shigella flexneri and 76 of Shigella sonnei, where only one S.sonnei isolate was resistant to CIP, but susceptible to TRV. About half the isolates of Salmonella spp. and S.sonnei and almost all S.flexneri isolates were resistant to ampicillin, and more than 60% of Shigella spp. isolates were resistant to trimethoprim-sulfamethoxazole. A 41% of Staphylococcus aureus and 55% of coagulase-negative staphylococci isolates were resistant to oxacillin, presenting a highly associated multi-resistance. The resistance to FQ was also strongly related to oxacillin resistance, but the resistance to TRV was significantly lower than the CIP resistance: 9% vs 57% for S.aureus and 4% vs 41% for coagulase-negative staphylococci. A similar behavior was observed with Enterococcus spp., where 54% of the isolates were resistant to norfloxacin and only 13% were resistant to TRV. Neither Streptococcus pneumoniae (n = 193) nor Haemophilus influenzae (n = 139) isolates presented resistant to TRV.

[Streptococcus pyogenes: penicillin and erythromycin susceptibility in the cities of Neuquen and Cipolletti]. / Streptococcus pyogenes: sensibilidad a penicilina y eritromicina en las ciudades de Neuquen y Cipolletti.

Penicillin resistance has not yet been detected in Streptococcus pyogenes. However macrolide-resistant streptococci have emerged in several countries. Only low rates of erythromycin-resistant S. pyogenes were reported in Argentina, with the exception of a 11.1% observed in Mendoza. The aim of the present study was to determine the susceptibility to penicillin and to erythromycin of 251 consecutive clinically-significant isolates of S. pyogenes obtained from four centers of Cipolletti and Neuquén during the period April-December 1998. The double disk test with erythromycin and clindamycin disks was employed as a screening method to detect ERY-resistant streptococci and to determine the phenotype of macrolide resistance. Disk diffusion was also employed for determining penicillin susceptibility. Macrolide-resistant isolates were also tested for penicillin, ceftriaxone, erythromycin, clindamycin and azithromycin susceptibility by the agar dilution method. Additionally they were also tested for erythromycin susceptibility by E-test (AB Biodisk, Solna, Sweden). All streptococci studied were susceptible to penicillin and thirty of them (12.0%) were resistant to erythromycin. All these resistant isolates were also resistant to azithromycin but susceptible to ceftriaxone and clindamycin. They showed the phenotype M (probably efflux-mediated mechanism) and the MICs of erythromycin ranged between 8 and 16 micrograms/ml. According to these results we conclude that in spite of universal susceptibility to penicillin in S. pyogenes, macrolide resistance is a matter of concern in Neuquén and Cipolletti. At least in those cities it appears to be necessary to routinely perform macrolide susceptibility tests in beta-hemolytic streptococci.

[Bacteremia caused by Enterococcus gallinarum with a high level of glycopeptide resistance: 1st documented cases in Argentina]. / Bacteriemia por Enterococcus gallinarum con alto nivel de resistencia a glicopéptidos: primer caso documentado en Argentina.

A case of bacteremia due to high-level-vancomycin- (MIC = 64 micrograms/ml) and high-level-teicoplanin- (MIC = 32 micrograms/ml) resistant Enterococcus gallinarum is described. Both genes, van C1 and van A, respectively conferring natural low-level resistance and acquired high-level resistance to vancomycin, were found in the enterococcal genoma. The present is the first report of an E. gallinarum isolate showing the van A genotype in Argentina.

[Consensus on antimicrobial sensitivity tests in gram-positive cocci. Subcommittee on Antimicrobials, SADEBAC (Argentinian Society of Clinical Bacteriology), Argentinian Association of Microbiology]. / Consenso sobre las pruebas de sensibilidad a los antimicrobianos en cocos gram-positivos.

Antimicrobial susceptibility testing is mainly performed in Argentina by disk diffusion method, following National Committee for Clinical Laboratory Standards (NCCLS) recommendations. We worked out new recommendations for the reporting and interpretation of this test when dealing with gram-positive cocci, in accordance to local trends and epidemiology. General considerations for performing the diffusion assay, quality control, and an update on susceptibility testing for gram-positive cocci are reported in this first document. The present update should be considered as a group of recommendations summarized by Argentinean experts and as the result of a consensus meeting coordinated by the Subcomisión de Antimicrobianos of the Sociedad Argentina de Bacteriología Clínica (Asociación Argentina de Microbiología). Experts in antimicrobial agents were convened in order to prepare this final document. These recommendations take into account local needs, affordability and availability to be used in current practice, tending to contribute to the correct antimicrobial treatment election, according to the particular microorganism and the infection sites.