Molecular epidemiology of methicillin-resistant Staphylococcus aureus from different population groups in Argentina.

OBJECTIVES: In Latin America, methicillin-resistantStaphylococcus aureus (MRSA) is a leading cause of nosocomial infections. Limited studies have addressed the molecular epidemiology of MRSA clones in Argentina, characterised by continuous human migratory movements. The aim of this study was to describe the MRSA epidemiology, including distinct patient populations from different regions of the country. METHODS: MRSA strains were collected in epidemiological studies conducted from 2009 to 2015 in three cities (Formosa, Córdoba and Tucumán) and involving four population groups: community adult patients; hospitalised adults; hospitalised children; and healthy children (nasal colonisation). Antimicrobial susceptibility testing, SCCmec and Panton-Valentine leukocidin (PVL) typing, pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were performed. RESULTS: A total of 120 MRSA isolates were recovered with an important population diversity in the groups studied; in community adult patients, MRSA isolates corresponded to ST5, ST267 and ST1619; from hospitalised adults they were ST97, ST5, ST72, ST125, ST200, ST647, ST747, ST935 and ST2941; from hospitalised children they were ST5, ST30, ST34, ST1163 and ST1619; and from colonised children they were ST5, ST125, ST34, ST100, ST1619, ST207 and ST1163. Results of SCCmec typing showed SCCmec I, SCCmec IIIA, SCCmec IV and SCCmec ND associated or not with PVL genes. CONCLUSIONS: MRSA genetic lineages have differing distribution in the three regions. The most prevalent was ST5 in colonisation, community and invasive settings. Here we describe ST34-SCCmec IV clone for the first time in the hospitalised paediatric population. These findings contribute to the understanding of epidemiological changes in recent years.

Multiple-time scaling and universal behavior of the earthquake interevent time distribution.

The interevent time distribution characterizes the temporal occurrence in seismic catalogs. Universal scaling properties of this distribution have been evidenced for entire catalogs and seismic sequences. Recently, these universal features have been questioned and some criticisms have been raised. We investigate the existence of universal scaling properties by analyzing a Californian catalog and by means of numerical simulations of an epidemic-type model. We show that the interevent time distribution exhibits a universal behavior over the entire temporal range if four characteristic times are taken into account. The above analysis allows us to identify the scaling form leading to universal behavior and explains the observed deviations. Furthermore, it provides a tool to identify the dependence on the mainshock magnitude of the c parameter that fixes the onset of the power law decay in the Omori law.

On-off intermittency in earthquake occurrence.

The clustered occurrence of earthquakes is viewed as an intermittent phenomenon, interpreting the clusters of events as chaotic bursts combined to the Poissonian occurrence of background seismicity. In particular, we suggest that it can be interpreted as an example of on-off intermittency. This kind of intermittency is parameter driven and exhibits certain universal statistical properties. The study of a Californian catalogue allows to interpret earthquake occurrence as an on-off intermittent phenomenon. Our results suggest the existence of a branching mechanism in earthquake occurrence well explained by epidemic type models.

Use of green fluorescent protein-based reporters to monitor balanced production of antifungal compounds in the biocontrol agent Pseudomonas fluorescens CHA0.

AIMS: To develop reporter constructs based on stable and unstable variants of the green fluorescent protein (GFP) for monitoring balanced production of antifungal compounds that are crucial for the capacity of the root-colonizing Pseudomonas fluorescens strain CHA0 to control plant diseases caused by soil-borne pathogenic fungi. METHODS AND RESULTS: Pseudomonas fluorescens CHA0 produces the three antifungal metabolites 2,4-diacetylphloroglucinol (DAPG), pyoluteorin (PLT) and pyrrolnitrin (PRN). The gfp[mut3] and gfp[AAV] reporter genes were fused to the promoter regions of the DAPG, PLT and PRN biosynthetic genes. The reporter fusions were then used to follow the kinetics of expression of the three antifungal metabolites in a microplate assay. DAPG and PLT were found to display an inverse relationship in which each metabolite activates its own biosynthesis while repressing the synthesis of the other metabolite. PRN appears not to be involved in this balance. However, the microbial and plant phenolic metabolite salicylate was found to interfere with the expression of both DAPG and PLT. CONCLUSIONS: The results obtained provide evidence that P. fluorescens CHA0 may keep the antifungal compounds DAPG and PLT at a fine-tuned balance that can be affected by certain microbial and plant phenolics. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge, the present study is the first to use stable and unstable GFP variants to study antibiotic gene expression in a biocontrol pseudomonad. The developed reporter fusions will be a highly valuable tool to study in situ expression of this bacterial biocontrol trait on plant roots, i.e. at the site of pathogen suppression.

[Distribution of capsular types and antimicrobial susceptibility of Streptococcus agalactiae causing infections in Argentina]. / Distribución de tipo capsular y sensibilidad antimicrobiana de Streptococcus agalactiae productores de infecciones en Argentina.

Streptococcus agalactiae is an endogenous bacterium that has emerged in the last 20 years as an etiological agent in both neonatal and perinatal infections, and in immunocompromised patients. The differentiation of the capsular polysaccharide, the presence of surface proteins c, X, R, and molecular methods allow classification in serotypes and genotypes. This identification is a useful tool for epidemiological purposes and virulence studies in this bacterium. The objective of this work was to study the serotypes and the antimicrobial susceptibility of isolates recovered from invasive diseases in different areas of Argentina. In the analyzed sample a fair predominance of Ia and III serotypes was recovered, followed by II and IV serotypes. All the isolates were found to be sensitive to penicillin. A 6% of resistance to erythromycin and a 4.5% to clindamycin were detected. In three of the isolates, constitutive MLS phenotype (resistance to macrolides, lincosamins and streptogramins) was founded, while in the remaining one, inducible MLS phenotype was detected. These results stress the importance of conducting a surveillance of the prevalent serotypes in our country with the goal of future prevention of this disease with an effective vaccine. The knowledge of the antimicrobial susceptibility profile will be also important to obtain therapeutic success in the treatment.

Distribución de tipo capsular y sensibilidad antimicrobiana de Streptococcusagalactiae productores de infecciones en Argentina / Distribution of capsular types and antimicrobial susceptibility of Streptococcus agalactiae causing infections in Argentina

Streptococcus agalactiae es una bacteria colonizante que ha emergido en los últimos años como causante de infecciones neonatales, perinatales y en pacientes con compromiso inmunológico. La caracterización del polisacárido capsular, de las proteínas de superficie (c, X, R), así como el análisis de marcadores moleculares, permiten su clasificación en serotipos y genotipos. Esto resulta de utilidad para fines epidemiológicos y para estudios de virulencia de la bacteria. El objetivo de este trabajo fue conocer los serotipos prevalentes y la sensibilidad antimicrobiana de aislamientos provenientes de procesos infecciosos en pacientes de distintas zonas de Argentina. En la muestra analizada se obtuvo predominio de los serotipos Ia y III, seguido de II y IV. Todas las cepas resultaron sensibles a penicilina. Se observó 6% de resistencia a eritromicina y 4,5% a clindamicina. En 3 de las cepas se detectó fenotipo MLS (resistencia a macrólidos, lincosaminas y estreptograminas) constitutiva y en una cepa, resistencia MLS inducible. Los resultados logrados en este estudio destacan la importancia de efectuar un relevamiento de los serotipos más frecuentes en nuestro país en vistas a la prevención de esta infección con una vacuna que realmente sea eficaz, como así también el conocimiento de la sensibilidad antimicrobiana para lograr éxito terapéutico en los tratamientos.

Streptococcus agalactiae is an endogenous bacterium that has emerged in the last 20 years as an etiological agent in both neonatal and perinatal infections, and in immunocompromised patients. The differentiation of the capsular polysaccharide, the presence of surface proteins c, X, R, and molecular methods allow classification in serotypes and genotypes. This identification is a useful tool for epidemiological purposes and virulence studies in this bacterium. The objective of this work was to study the serotypes and the antimicrobial susceptibility of isolates recovered from invasive diseases in different areas of Argentina. In the analyzed sample a fair predominance of Ia and III serotypes was recovered, followed by II and IV serotypes. All the isolates were found to be sensitive to penicillin. A 6% of resistance to erythromycin and a 4.5% to clindamycin were detected. In three of the isolates, constitutive MLS phenotype (resistance to macrolides, lincosamins and streptogramins) was founded, while in the remaining one, inducible MLS phenotype was detected. These results stress the importance of conducting a surveillance of the prevalent serotypes in our country with the goal of future prevention of this disease with an effective vaccine. The knowledge of the antimicrobial susceptibility profile will be also important to obtain therapeutic success in the treatment.

Erythromycin-resistant Streptococcus pyogenes in Argentina

Erythromycin (ERY) resistance in Streptococcus pyogenes has recently emerged as a problem of growing concern all through the world. We are presenting the comparison of results of the continuous surveillance of erythromycin resistance in S. pyogenes performed since 1989 in the Hospital de Pediatría J.P.Garrahan of Buenos Aires City, with independently observed rates in other five centers of Buenos Aires and seven centers of six other Argentinian cities, obtained between 1999 and 2001. A significant increase of erythromycin resistance was observed among S. pyogenes isolated in the Hospital Garrahan (6.6% in 1998-1999 to 9.9% in 2000). Similar trends were also detected in other centers of other Argentinian cities when recent data were compared to results of a multicenter study performed in 1995. However, lower rates of resistance were recorded in Mendoza, Cipolletti and Neuquén in comparison with data of 1995, 1998 and 1998 respectively. The reason of such decreasing resistance rates deserves to be investigated. The average of ERY-resistance rates obtained in the surveyed centers was 6.7% (range 0.5-14.1%). Control of antimicrobial use should be performed to warrant the future effectiveness of macrolide antibiotics regarding the positive association between use and resistance. These results also suggest that susceptibility tests for macrolides should be performed whenever S. pyogenes is isolated in Argentina.

La resistencia a la eritromicina en Streptococcus pyogenes ha emergido en los últimos tiempos como un problema creciente en todo el mundo. En este trabajo se presenta la comparación de los resultados de la vigilancia continua de la resistencia a la eritromicina que se viene realizando en el Hospital de Pediatría J.P.Garrahan de Buenos Aires desde 1989, con resultados independientes de otros cinco centros de la ciudad de Buenos Aires y siete de otras seis ciudades argentinas, obtenidos entre 1999 y 2001. Se observó un aumento significativo en el Hospital Garrahan (6.6% en1998-1999 a 9.9% en el año 2000) y una tendencia similar en otros centros de diversas ciudades argentinas si secomparan estos datos con los de un estudio multicéntrico realizado en 1995. No obstante, se registraron menoresporcentajes de resistencia en Mendoza, Neuquén y Cipolletti, en relación a lo hallado en 1995, 1998 y 1998respectivamente. La razón de esta disminución merece ser investigada. El porcentaje promedio de resistencia aeritromicina obtenido en los distintos centros participantes de este estudio fue de 6.7% (rango 0.5-14.1%). Debeefectuarse un control en el uso de estos antibióticos para garantizar la efectividad futura de los macrólidos, teniendo en cuenta la asociación estrecha entre uso y resistencia. Estos resultados sugieren que deberían realizarse pruebas de sensibilidad a los macrólidos para todos los aislamientos de S. pyogenes en la Argentina.

Acid exposure and altered acid clearance in GERD patients treated for Helicobacter pylori infection.

BACKGROUND: After the eradication of Helicobacter pylori, an increased incidence of gastroesophageal reflux disease and acid gastric secretion have been reported. AIM: To evaluate the effect of Helicobacter pylori-eradication on proximal and distal gastroesophageal reflux and acid clearance in patients with gastroesophageal reflux disease. PATIENTS AND METHODS: Sixty-eight gastroesophageal reflux disease patients (age range 18-61 years) were studied by upper endoscopy. All underwent esophageal manometry and dual probe 24-h pH-metry. RESULTS: Percent of time at pH<4 was significantly increased in the proximal esophagus of Helicobacter pylori-eradicated patients compared to Helicobacter pylori-negative (2.4+/-0.5 vs. 1.0+/-0.2; p<0.01); no differences were found in the distal esophagus (14.0+/-3.7 vs. 9.0+/-1.4%, NS). The total number of reflux episodes was significantly higher in the proximal oesophagus of Helicobacter pylori-eradicated patients (37+/-3 vs. 22+/-3, p<0.05). In the distal esophagus, acid clearance was significantly longer, both during total time (1.4+/-0.2 vs. 0.8+/-0.7 min, p<0.01), and in the supine period (8.5+/-2.7 vs. 2.7+/-0.4 min, p<0.05). No differences were reported in the manometric parameters of the two groups of patients. CONCLUSION: In patients with gastroesophageal reflux disease, Helicobacter pylori eradication is associated with increased acid exposure of the proximal esophagus and delayed distal acid clearance.

[Group B streptococcal infections in adults, excluding genital infections]. / Infecciones por estreptococos grupo B en adultos excluyendo infecciones genitales.

Group B streptococcus (GBS) or Streptococcus agalactiae is recognized as a mayor cause of neonatal meningitis, sepsis and infections during pregnancy. However, in recent years there have been several reports concerning GBS infections in non pregnant adult population, specially in immunocompromised hosts and in patients with severe underlying diseases such as diabetes mellitus and cancer. We report a series of 45 cases which occurred in nonpregnant adult population during a period of two years. The average age was 50.8 years and most patients (38/44) had one or more risk factors: diabetes mellitus was the most significant underlying disease. The most frequent infection localization was skin and soft tissues followed by urinary tract infection. Several isolated cases of pneumonia, bacteremia, endocarditis, endometritis and peritonitis were observed. GBS infections should no longer be exclusively considered as perinatal and peripartum events. New clinical presentations are arising in non pregnant adult population with special incidence in immunocompromised hosts. We are obliged to keep this in mind and remember that SGB may be a possible etiologic agent for infections, particularly in skin and soft tissues of diabetic patients.

Infecciones por estreptococos grupo B en adultos excluyendo infecciones genitales / [Group B streptococcal infections in adults, excluding genital infections]

Group B streptococcus (GBS) or Streptococcus agalactiae is recognized as a mayor cause of neonatal meningitis, sepsis and infections during pregnancy. However, in recent years there have been several reports concerning GBS infections in non pregnant adult population, specially in immunocompromised hosts and in patients with severe underlying diseases such as diabetes mellitus and cancer. We report a series of 45 cases which occurred in nonpregnant adult population during a period of two years. The average age was 50.8 years and most patients (38/44) had one or more risk factors: diabetes mellitus was the most significant underlying disease. The most frequent infection localization was skin and soft tissues followed by urinary tract infection. Several isolated cases of pneumonia, bacteremia, endocarditis, endometritis and peritonitis were observed. GBS infections should no longer be exclusively considered as perinatal and peripartum events. New clinical presentations are arising in non pregnant adult population with special incidence in immunocompromised hosts. We are obliged to keep this in mind and remember that SGB may be a possible etiologic agent for infections, particularly in skin and soft tissues of diabetic patients.

Role of diaphragmatic crura and lower esophageal sphincter in gastroesophageal reflux disease: manometric and pH-metric study of small hiatal hernia.

The rapid pull-through (RPT) technique during esophageal manometry helps to identify various pressure profiles of hiatal hernia (HH), based on the presence of two high pressure zones: the diaphragmatic crura (DC) and the lower esophageal sphincter (LES). Our aim was to correlate different HH profiles with frequency of reflux episodes in patients with gastroesophageal reflux disease (GERD). Seventy-eight patients with GERD and HH underwent esophageal manometry with RPT and were grouped according to the prevalent pressure profile of HH. Twenty-four-hour pH-metry served to quantify traditional (TR) and nontraditional refluxes (drop of 1 pH unit with pH > 4 or pH < 4 and time < 5 sec) (NTR) during total, upright, and recumbent periods. The group with a prevalent "flat" HH profile, representing LES and DC impairment, had significantly more TRs in total time of reflux (P < 0.01) and in recumbent and upright periods (P < 0.05) compared to the group with a prevalence of the two pressure peaks, corresponding to LES and DC efficiency. However, the group with the flat profile had significantly more NTRs + TRs than the group with pressure peaks in total time (P < 0.01) and recumbent position (P < 0.001) but not in the upright position. Hiatal hernia predisposes to GERD, but only the associated impairment of the LES and diaphragmatic crura pressures represents a condition of high risk for gastroesophageal reflux events.

Manometric study of hiatal hernia and its correlation with esophageal peristalsis.

The pathogenesis of gastroesophageal reflux disease (GERD) is considered multifactorial, but alterations of the esophagogastric junction (EGJ) and hiatal hernia play a prominent role. The correlations between hiatal hernia and the other pathogenetic factors are as yet unclear, and they need to be investigated by a methodological approach based on new anatomic and functional criteria. Our aim was to study, by stationary manometry, the relationships between small reducible hiatal hernia, identified by endoscopy, and esophageal peristalsis, in patients with and without GERD. According to the absence or presence of esophagitis (E), and the absence or presence of hiatal hernia (H), 58 subjects were divided into four groups: controls 10; H 14; E 10; and HE 24. Stationary manometry was performed by the rapid pull-through (RPT) technique, with catheter water perfused, to study the lower esophageal high pressure zone [lower esophageal sphincter (LES) and diaphragmatic crura] and the parameters of esophageal peristalsis. In patients with hiatal hernia, the various combinations of peak and/or deflection of manometric line pressure identified five EGJ profiles, only one of which reveals (by one-peak profile due to superimposed LES and diaphragmatic crura) the reducibility of the hernia. The frequency of the five profiles was calculated in the HE and H groups: a two-peak profile was significantly more prevalent in these patients, although less so in the group with esophagitis. In E patients the distal amplitude and the distal propagation of esophageal waves were significantly lower than in the other three groups (P < 0.05 vs controls and group HE; P < 0.01 vs group H). Furthermore, the distal amplitude was significantly higher in the group H than in the HE (P < 0.01). Our results show a better definition of hiatal hernia morphology, via the RPT technique, disclosing five pressure profiles. In addition, a significant link was found between small reducible hiatal hernia without GERD and wave amplitude of the distal esophagus. The amplification of peristaltic clearing may be considered the initial protective process against acid reflux; the breakdown of this mechanism may trigger the pathological sequence of GERD.

Monoethylglycinexylidide production as a measure in predicting hepatic histology.

To clarify whether monoethylglycinexylidide (MEGX) production can be useful in predicting the severity of chronic liver disease, 51 subjects were enrolled in this study: 13 mild-moderate CAH (group A), 9 severe CAH (group B), 29 cirrhosis, 18 compensated and 11 decompensated disease (group C). The patients were injected with a 1 mg/Kg lidocaine bolus i.v. and serum-sampled for MEGX values (time 15, 30 and 60 minutes), determined by TDX-immunoassay. MEGX formation was significantly different (Mann-Whitney U test) in the three groups at each time interval, especially at 30 min: group A = 70.5 +/- 9.9 ng/ml (mean +/- SD); group B = 49.7 +/- 7.2; group C = 37.2 +/- 19.5 (in A vs B, p = 0.0003; in A vs C, p < 0.0001; in B vs C, p = 0.0237). The difference between compensated and decompensated cirrhosis was always significant (p = 0.0099, 0.0005, 0.0113 respectively) but between severe CAH and compensated cirrhosis it was marginal only at 15min (p = 0.0763) and absent at 30 and 60min. At 30min MEGX values > 60 suggest mild-moderate CAH, < 40 cirrhosis, < 30 decompensated cirrhosis, between 40 and 60 severe CAH or compensated cirrhosis. We are of the opinion that the MEGX test could be utilized to predict hepatic histology.