The DATAC study: a new growth database. Description of the epidemiology, diagnosis and therapeutic attitude in a group of Spanish children with short stature.

BACKGROUND: Recombinant human growth hormone (rhGH) availability has allowed the treatment of a greater number of growth disorders. It is important to get an insight into the clinical characteristics of the paediatric population before rhGH treatment. METHODS: An observational, retrospective and multicentre study was conducted to evaluate the patients' baseline characteristics prior to rhGH therapy. RESULTS: A total of 1404 patients (53.8% males) aged 0.5-17.3 years were included. Clinical conditions were as follows: GH deficiency (GHD), 66.0%; small for gestational age (SGA), 29.7%; and Turner syndrome (TS), 4.3%. Male gender was predominant in most growth disorders; age at diagnosis was higher in GHD patients; therapy with rhGH started at lower chronological age in SGA and TS groups. CONCLUSION: The baseline characteristics of the population to be treated with rhGH were similar to those reported in other growth databases. Delayed age at treatment initiation should increase the awareness of these disorders among general paediatricians and entice them to refer children suspected of having these disorders to a specialist.

Adverse events during the titration phase of interferon-beta in remitting-relapsing multiple sclerosis are not predicted by body mass index nor by pharmacodynamic biomarkers.

BACKGROUND: This study aimed to correlate body mass index or biomarkers with the frequency of common adverse events (AEs) with subcutaneous IFN ß-1a during treatment titration in patients with relapsing-remitting multiple sclerosis previously naïve to IFN ß. METHODS: Eighty-four patients (66.3% females) were followed up during 8 weeks, 25.3% were overweight and 14.5% were obese. RESULTS: Biomarkers steadily increased during all study period by 45.3% for ß2-microglobulin, 262.8% for olygoadenylate synthetase-1, and 92.8% for neopterin. Overall AE reporting did not vary with the dose or treatment duration. CONCLUSIONS: BMI was not predictive of increased risk for AEs. Biomarkers did not discriminate on the frequency of any AE either.

Histological approach to Bacillus subtilis colony-biofilm: evolving internal architecture and sporulation dynamics.

Bacillus subtilis has been used as a classic model to study biofilm formation and sporulation process. Colonies of wild-type strains usually have a complex external morphology, but the details of their internal architecture are still undisclosed. Since bacterial biofilms fulfill the criteria to be considered tissues, the aim of this work was to analyse B. subtilis colony-biofilm internal architecture evolution and sporulation dynamics using histological techniques. Transversal sections of colony-biofilms incubated from 24 hours up to 20 days were stained using histochemical techniques to analyse the internal structure by light and electron microscopy. A morphometric study of the different structural biofilm components was performed by image analysis, and an application to quantify spores was developed. Internal biofilm architecture was characterised by a stratified pattern, which evolved from 3 strata at 24 hours, up to 5 strata at 20 days. At 48 hours, strata at the central area of the biofilm was folded, resulting in elevated structures (vein-like structures) that could reach up to 465 µm in height. Sporulation started at 48 hours, at the top of the vein-like structures, at the interface between the two uppermost strata. At 20 days spores formed a continuous central layer, representing 7.5% of the total biofilm. In summary, our results demonstrate that B. subtilis colony-biofilm has a complex and organized internal architecture, evolving over time, and taking place in different cell subpopulations with different functionalities. Furthermore, in situ spore quantification described in this work could be a good alternative to the classical chamber counting.

Invasion of solid culture media: a widespread phenotypic feature of clinical bacterial isolates.

OBJECTIVES: The in-depth growth in solid culture media is a common feature in filamentous fungi and yeasts. However, there are very few bacterial species in which this phenomenon has been documented. The aim of this work was to assess the agar invasiveness of a wide range of Gram-positive and Gram-negative bacterial species of clinical interest. MATERIAL AND METHODS: Three different clinical isolates for each of eleven bacterial species were plated onto Columbia blood agar and let grow up to 15 days. Isolated colonies were processed by histological methods, embedded in epoxy resin, and then, semithin sections were stained with toluidine blue and visualized by light microscopy. RESULTS: Growth within the agar was observed in at least one strain in 9 of the 11 studied species. Invasions of Gram-negative rods were small, not plentiful, and round or triangle-shaped. In Gram-positive cocci, invasions were of big size, abundant and of variable shape (lentiform, globular, irregular, arrowhead) depending on the species. CONCLUSIONS: We propose that the growth within the agar can indicate a survival strategy common to many bacterial species, and so far, not previously reported. This strategy could be either a nutrient gradient tropism or the spread and colonization of new ecological niches, with potential implications in pathogeny.

Invasion of solid culture media: a widespread phenotypic feature of clinical bacterial isolates

Objetivos. El crecimiento en profundidad en medios de cultivo sólidos es un fenómeno habitual en hongos filamentosos y levaduras. Sin embargo, son muy escasas las especies bacterianas en las que se ha documentado este fenómeno. El objetivo de este trabajo fue comprobar la capacidad de invasión del agar de un amplio abanico de especies bacterianas grampositivas y gramnegativas de interés clínico. Material y métodos. Se cultivaron tres cepas de cada una de once especies bacterianas sobre agar Columbia hasta un máximo de 15 días. Colonias aisladas fueron procesadas e incluidas en resina epoxi por métodos histológicos y secciones transversales semifinas teñidas con azul de toluidina fueron visualizadas por microscopía óptica. Resultados. Se observó crecimiento en el interior del agar en al menos una de las cepas de nueve de las especies estudiadas. En bacilos gramnegativos, las invasiones eran escasas en número, pequeñas y con morfología redondeada o triangular. En cocos grampositivos las invasiones eran de gran tamaño, numerosas y de morfología variable (lentiforme, globular, irregular, en forma de punta de flecha, etc.) según la especie. Conclusiones. En nuestra opinión, el crecimiento en el interior del agar puede significar una estrategia de supervivencia común a muchas especies bacterianas y hasta ahora prácticamente no descrita. Esta estrategia podría ser el reflejo de un tropismo a favor de gradiente de nutrientes, o bien un fenómeno de diseminación y colonización de nuevos nichos ecológicos, con posibles implicaciones en la patogenia(AU)

Objectives. The in-depth growth in solid culture media is a common feature in filamentous fungi and yeasts. However, there are very few bacterial species in which this phenomenon has been documented. The aim of this work was to assess the agar invasiveness of a wide range of Gram-positive and Gram-negative bacterial species of clinical interest. Material and methods. Three different clinical isolates for each of eleven bacterial species were plated onto Columbia blood agar and let grow up to 15 days. Isolated colonies were processed by histological methods, embedded in epoxy resin, and then, semithin sections were stained with toluidine blue and visualized by light microscopy. Results. Growth within the agar was observed in at least one strain in 9 of the 11 studied species. Invasions of Gramnegative rods were small, not plentiful, and round or triangleshaped. In Gram-positive cocci, invasions were of big size, abundant and of variable shape (lentiform, globular, irregular, arrowhead) depending on the species. Conclusions: We propose that the growth within the agar can indicate a survival strategy common to many bacterial species, and so far, not previously reported. This strategy could be either a nutrient gradient tropism or the spread and colonization of new ecological niches, with potential implications in pathogeny(AU)

Discordancia en los resultados de sensibilidad mediante método de difusión en agar de aislados de Pseudomonas aeruginosa en pacientes con fibrosis quística tras pre-incubación anaerobia y su potencial relevancia clínica / Discrepancy in the disk diffusion susceptibility test of Pseudomonas aeruginosa strains isolated from cystic fibrosis patients after anaerobic preincubation and its potential clinical relevance

Introducción. En la fibrosis quística las células de Pseudomonas aeruginosa crecen en el interior de la espesa mucosidad, y pese a ser un organismo aerobio estricto, se desarrolla en un ambiente donde la presión de oxígeno es muy limitada. Posibles movimientos de la masa mucosa podrían dejar expuestas de forma súbita a las células de P. aeruginosa a concentraciones altas de oxígeno. El objetivo del estudio fue determinar cómo afecta a la sensibilidad antibiótica de P. aeruginosa un período de incubación anaerobia. Material y métodos. Se emplearon 4 cepas de P. aeruginosa (NTCC 23389 y 3 aislados clínicos). Para la determinación de la sensibilidad antibiótica se empleó el método de difusión en agar. Resultados. La preincubación anaerobia produce cambios en la sensibilidad en todas las cepas estudiadas. Todas las cepas sensibles en aerobiosis resultaron también sensibles tras la incubación anaerobia a excepción de la cepa 2 para los betalactámicos. El tipo de respuesta resulta dependiente de cada cepa, siendo especialmente significativo el incremento en la sensibilidad observado en el caso de ciprofloxacino para dos de los tres aislados clínicos. Conclusiones. La sensibilidad de cepas P. aeruginosa varía si han sido previamente expuestas a condiciones de anaerobiosis. Tratamientos que favorezcan la fluidificación de la mucosidad podrían contribuir aumentando el éxito del tratamiento con ciertos antibióticos(AU)

Introduction. In cystic fibrosis, the Pseudomonas aeruginosa cells grow inside the thick mucus layer. In spite of being an obligate aerobe, P. aeruginosa is able to grow in a limited oxygen environment. Bacterial cells could be suddenly exposed to high oxygen levels due to the movements of the mucus mass. The aim of study was to determine the impact of a previous anaerobic incubation on the antimicrobial susceptibility of P. aeruginosa strains isolated from patients with cystic fibrosis. Materials and Methods. Four P. aeruginosa strains were used in this study (ATCC 23389 and 3 clinical isolates). The disk diffusion method was used to determine the antimicrobial susceptibility. Results. The anaerobic pre-incubation produced changes on the susceptibility in all studied strains. All susceptible strains after an aerobic incubation remained susceptible after an anaerobic incubation except one clinical strain, which became resistant to betalactams. The response was strain-dependent and the most significant increase in susceptibility was observed in two of the three clinical isolates when ciprofloxacin was used. Conclusions. The antimicrobial susceptibility of P. aeruginosa strains varies after their exposure to anaerobic conditions. Treatments promoting mucus fluidization could contribute to increase the antimicrobial efficacy(AU)

[Discrepancy in the disk diffusion susceptibility test of Pseudomonas aeruginosa strains isolated from cystic fibrosis patients after anaerobic preincubation and its potential clinical relevance]. / Discordancia en los resultados de sensibilidad mediante método de difusión en agar de aislados de Pseudomonas aeruginosa en pacientes con fibrosis quística tras pre-incubación anaerobia y su potencial relevancia clínica.

INTRODUCTION: In cystic fibrosis, the Pseudomonas aeruginosa cells grow inside the thick mucus layer. In spite of being an obligate aerobe, P. aeruginosa is able to grow in a limited oxygen environment. Bacterial cells could be suddenly exposed to high oxygen levels due to the movements of the mucus mass. The aim of study was to determine the impact of a previous anaerobic incubation on the antimicrobial susceptibility of P. aeruginosa strains isolated from patients with cystic fibrosis. MATERIALS AND METHODS: Four P. aeruginosa strains were used in this study (ATCC 23389 and 3 clinical isolates). The disk diffusion method was used to determine the antimicrobial susceptibility. RESULTS: The anaerobic pre-incubation produced changes on the susceptibility in all studied strains. All susceptible strains after an aerobic incubation remained susceptible after an anaerobic incubation except one clinical strain, which became resistant to betalactams. The response was strain-dependent and the most significant increase in susceptibility was observed in two of the three clinical isolates when ciprofloxacin was used. CONCLUSIONS: The antimicrobial susceptibility of P. aeruginosa strains varies after their exposure to anaerobic conditions. Treatments promoting mucus fluidization could contribute to increase the antimicrobial efficacy.

El papel de la industria farmacéutica. ¿Por qué no se comercializan nuevos antibióticos? / The role of the pharmaceutical industry. Why aren't new antibiotics being marketed?

El escaso o nulo interés social y político en el problema de las resistencias a antibióticos, la dificultad en la identificación de moléculas activas frente a nuevas dianas, pero, sobre todo, su baja rentabilidad relativa frente a otras clases de fármacos, así como la incertidumbre y arbitrariedad por parte de las autoridades regulatorias a la hora de evaluar su eficacia, están en la base de la enorme reducción en el ritmo de comercialización de nuevos antibióticos. Las condiciones actuales no favorecen la inversión en nuevos antibióticos por parte de la industria farmacéutica, que tiene a su disposición áreas terapéuticas con mucha mayor rentabilidad potencial y otros problemas propios de los que ocuparse. Ya que no se puede forzar a la industria a desarrollar antibióticos habrá que implementar lo antes posible políticas que estimulen su interés en ellos, o bien encontrar la manera de que los diferentes estados y autoridades sustituyan a la industria farmacéutica en esta tarea

The lack or absence of social and political interest in the problem of antibiotic resistance, the difficulty in identifying active molecules against new targets, and above all, low profitability in comparison to other types of drugs, as well as uncertainty and the arbitrary nature of regulatory authorities in terms of assessing effectiveness, all contribute to a significant slowdown in the marketing of new antibiotics. Current conditions do not favor investment in new antibiotics by the pharmaceutical industry, which has available therapeutic areas with far greater profit potential, and other problems of its own to handle. Since we cannot force the industry to develop antibiotics, it is necessary to implement policies as soon as possible that stimulate interest in developing them, or find a way for the states and regulatory authorities to replace the pharmaceutical industry in this task

Recomendaciones en el tratamiento antibiótico empírico de la infección intraabdominal / Recommendations in the empiric anti-infective agents of intra-abdominal infection

Un número importante de pacientes con infección intraabdominal desarrollan estados avanzados de la infección y la mortalidad es todavía superior al 20%. El fracaso es multifactorial y se relaciona con el incremento de resistencias bacterianas, el tratamiento empírico inapropiado, la mayor comorbilidad de los pacientes y el mal control del foco de infección. Estas guías analizan cada uno de estos problemas y proponen medidas para evitar el fracaso, basadas en la major evidencia científica actual (AU)

A significant number of patients with abdominal infection develop advanced stages of infection and mortality is still above 20%. Failure is multifactorial and is associated with an increase of bacterial resitance, inappropriate empirical treatment, a higher comorbidity of patients and poor source control of infection. These guidelines discuss each of these problems and propose measures to avoid the failure based on the best current scientific evidence (AU)

Multicenter study evaluating the role of enterococci in secondary bacterial peritonitis.

A 1-year prospective multicenter study was performed to explore the significance of the presence of enterococci in cultures of peritoneal fluid from patients with secondary bacterial peritonitis in seven Spanish hospitals. The clinical records of patients with positive peritoneal fluid cultures were reviewed and distributed into cases (patients with cultures yielding enterococci) and controls (patients with cultures not yielding enterococci). Of a total of 158 records, 38 (24.1%) were cases and 120 (75.9%) were controls. The percentages or the scores (cases versus controls) for the variables included in the multivariate analysis were as follows: age of >50 years, 89.5% versus 68.3%; malignancy, 39.5% versus 18.3%; chronic obstructive pulmonary disease (COPD), 15.8% versus 4.2%; postoperative peritonitis, 55.3% versus 30.1%; nosocomial onset, 57.9% versus 34.2%; a higher Charlson comorbidity index, 3.29 +/- 3.38 versus 1.84 +/- 2.31; APACHE II score, 10.71 +/- 4.37 versus 8.76 +/- 5.49; ultimately or rapidly fatal disease, 63.2% versus 34.8%; need for surgical reintervention, 36.1% versus 15.1%; and admission to an intensive care unit, 45.9% versus 30.8%. In the multivariate analysis, enterococci were associated only with postoperative peritonitis (P = 0.009; odds ratio [OR] = 5.0; 95% confidence interval [CI] = 1.49 to 16.80), a higher Charlson comorbidity index (P = 0.002; OR = 1.30; 95% CI = 1.11 to 1.54), and COPD (P = 0.046; OR = 6.50; 95% CI = 1.04 to 40.73). The results of this study showed that enterococci were associated with comorbidity. An association with mortality could not be demonstrated.

[Recommendations in the empiric anti-infective agents of intra-abdominal infection]. / Recomendaciones en el tratamiento antibiótico empírico de la infección intraabdominal.

A significant number of patients with abdominal infection develop advanced stages of infection and mortality is still above 20%. Failure is multifactorial and is associated with an increase of bacterial resitance, inappropriate empirical treatment, a higher comorbidity of patients and poor source control of infection. These guidelines discuss each of these problems and propose measures to avoid the failure based on the best current scientific evidence.

[The role of the pharmaceutical industry. Why aren't new antibiotics being marketed?]. / El papel de la industria farmacéutica. ¿Por qué no se comercializan nuevos antibióticos?

The lack or absence of social and political interest in the problem of antibiotic resistance, the difficulty in identifying active molecules against new targets, and above all, low profitability in comparison to other types of drugs, as well as uncertainty and the arbitrary nature of regulatory authorities in terms of assessing effectiveness, all contribute to a significant slowdown in the marketing of new antibiotics. Current conditions do not favor investment in new antibiotics by the pharmaceutical industry, which has available therapeutic areas with far greater profit potential, and other problems of its own to handle. Since we cannot force the industry to develop antibiotics, it is necessary to implement policies as soon as possible that stimulate interest in developing them, or find a way for the states and regulatory authorities to replace the pharmaceutical industry in this task.

In vitro effect of physiological concentrations of human albumin on the antibacterial activity of tigecycline.

OBJECTIVES: To determine C(max) tigecycline activity in the presence/absence of physiological concentrations of human albumin with free fraction concentrations as controls. METHODS: Killing curves (final inoculum: 1.0-5.0 x 10(7) cfu/mL) were performed with 0.88 mg/L final concentrations (serum C(max) after a 100 mg 1 h infusion) in Mueller-Hinton broth supplemented with Ca(2+) and Mg(2+) (MH) and in MH with 4 g/dL human albumin. Controls were curves in MH with concentrations similar to the free fraction (fC(max) = 0.17 mg/L) calculated using protein binding. Activity was measured as log(10) initial inoculum reduction (log(10) initial inoculum-log(10) at 12 h/24 h). Target strains (tigecycline MIC/MBC; mg/L) were: methicillin-resistant Staphylococcus aureus heteroresistant to vancomycin (0.12/0.25); Enterococcus faecium (0.12/0.25); Escherichia coli producing extended-spectrum beta-lactamase (0.12/0.25); and Acinetobacter baumannii (0.25/1). RESULTS: At 24 h the fC(max) produced mean decreases of < or =0.1 cfu/mL for all strains, in contrast to the bactericidal activity (mean >3 log(10) reduction) provided by C(max) concentrations in the presence or absence of albumin for E. coli and E. faecium, and an activity nearly bactericidal for S. aureus (mean approximately 2.8 log(10) reduction). In the case of the A. baumannii isolate the C(max) in the presence or absence of albumin produced a mean reduction of 2.56 log(10) cfu/mL at 12 h (time of one dosing interval), with a bacteriostatic profile when considering 24 h colony counts (similar counts at 0 and 24 h). CONCLUSIONS: Correcting the total concentration for the reported literature binding values is unreliable since tigecycline antibacterial activity was greater than that suggested by the free fraction of the drug.

Evaluation of the ability of erythromycin-resistant and -susceptible pharyngeal group A Streptococcus isolates from Spain to enter and persist in human keratinocytes.

The macrolide-resistance rate among group A Streptococcus (GAS) isolates has increased in some European countries. To investigate the reasons for this increase, the ability of 60 erythromycin-resistant and 61 erythromycin-susceptible, genetically unrelated, pharyngeal GAS isolates from Spain to enter and persist within human keratinocytes was evaluated. It was observed that erythromycin resistance was associated with the presence of prtF1, a gene related to invasiveness, whereas no association was observed with the ability to enter human keratinocytes. However, the ability to enter human keratinocytes was not associated with the presence of prtF1 or with the emm type, suggesting that interaction with keratinocytes depends on several characteristics of the isolate. Almost all strains (95.9 %) were capable of persisting within human keratinocytes. However, most of them (91.7 %) exhibited a decline in viability over time. Interestingly, the ability to persist within keratinocytes protected from the action of the beta-lactams was higher among erythromycin-resistant isolates and correlated with their ability to avoid the induction of cellular lysis. These observations suggest that if the carrier state results from intracellular GAS survival, the association between erythromycin resistance and intracellular persistence may represent a serious problem for the eradication of these isolates.

Ampicillin-resistant non-beta-lactamase-producing Haemophilus influenzae in Spain: recent emergence of clonal isolates with increased resistance to cefotaxime and cefixime.

The sequence of the ftsI gene encoding the transpeptidase domain of penicillin-binding protein 3 (PBP 3) was determined for 354 nonconsecutive Haemophilus influenzae isolates from Spain; 17.8% of them were ampicillin susceptible, 56% were beta-lactamase nonproducing ampicillin resistant (BLNAR), 15.8% were beta-lactamase producers and ampicillin resistant, and 10.4% displayed both resistance mechanisms. The ftsI gene sequences had 28 different mutation patterns and amino acid substitutions at 23 positions. Some 93.2% of the BLNAR strains had amino acid substitutions at the Lys-Thr-Gly (KTG) motif, the two most common being Asn526 to Lys (83.9%) and Arg517 to His (9.3%). Amino acid substitutions at positions 377, 385, and 389, which conferred cefotaxime and cefixime MICs 10 to 60 times higher than those of susceptible strains, were found for the first time in Europe. In 72 isolates for which the repressor acrR gene of the AcrAB efflux pump was sequenced, numerous amino acid substitutions were found. Eight isolates with ampicillin MICs of 0.25 to 2 microg/ml showed changes that predicted the early termination of the acrR reading frame. Pulsed-field gel electrophoresis analysis demonstrated that most BLNAR strains were genetically diverse, although clonal dissemination was detected in a group of isolates presenting with increased resistance to cefotaxime and cefixime. Background antibiotic use at the community level revealed a marked trend toward increased amoxicillin-clavulanic acid consumption. BLNAR H. influenzae strains have arisen by vertical and horizontal spread and have evolved to adapt rapidly to the increased selective pressures posed by the use of oral penicillins and cephalosporins.

Impact of amoxicillin, associated or not with clavulanic acid, on pharyngeal colonization and selection of Streptococcus pneumoniae resistance in children under 5 years of age.

Among young children, pneumococcal nasopharyngeal colonization (NPC) rates of >90% have been described. The aim of our study was to assess the effect of amoxicillin exposure on the NPC. From Dec 2001 to Feb 2004, less than 5 years old children with respiratory symptoms and fever who were prescribed amoxicillin were eligible. Three nasopharyngeal swabs were taken: at the time of the initial visit (IV), 60 hours after amoxicillin discontinuation (end of treatment visit, ETV), and 4 weeks later (follow-up visit, FUV). One hundred and thirty four children were included. NPC was detected in 58.5%, 42.9% and 51% of <1, 1-2 and >2 years-old children respectively (NS). Vaccine serotypes (VS) or vaccine-related serotypes (VRS) were identified in 80%, 40% and 55% of <1-year-old, 1-2 year-old and >2-year-old children respectively (NS). The proportion of PNSSP was 60% in <1-year-old children, 43% in 1-2 year-old children and 40% in >2-year-old children (NS). 49 out of 134 (36.5%) children completed the three study visits. 51%, 22.4% and 46.9% of those were colonized at IV, ETV and FUV, respectively (p=0.007). The percentage of resistant SP was 28%, 45.5% and 8.7% (p=0.05) for penicillin. In children <1 year of age, a higher proportion of SP colonization, presence of VS and PNSSP was found. A downfall of NPC at the end of therapy was observed. NPC returned to baseline levels thanks to "de novo" colonization in half of the cases, a few weeks after.

Geographical and ecological analysis of resistance, coresistance, and coupled resistance to antimicrobials in respiratory pathogenic bacteria in Spain.

A multicenter susceptibility surveillance (the S.A.U.C.E. project) including 2,721 Streptococcus pneumoniae, 3,174 Streptococcus pyogenes, and 2,645 Haemophilus influenzae consecutive isolates was carried out in 25 hospitals all over Spain from November 2001 to October 2002 to evaluate the current epidemiology of resistance of the main bacteria involved in community-acquired respiratory tract infections. Susceptibility testing was performed in a single centralized laboratory by a broth microdilution method. The prevalence of resistant S. pneumoniae strains was 0.4% for cefotaxime, 4.4% for amoxicillin and amoxicillin-clavulanic acid, 25.6% for cefuroxime-axetil, 34.5% for erythromycin, clarithromycin, and azithromycin, and 36.0% for cefaclor. Phenotypes of resistance to erythromycin were MLS(B) (macrolide-lincosamide-streptogramin B) in 89.9% (gene ermB) and M (macrolide) in 9.7% of cases (gene mefA). No strain harbored both genes simultaneously. Serotypes 19, 6, 23, 14, and 3 were the most prevalent, accounting for 54.6% of the total isolates. Resistance to macrolides seems to be the most alarming point, since among penicillin-susceptible isolates it reached 15.1% compared to 55.8% among penicillin-resistant strains. Geographically, a number of regions had rates of erythromycin resistance above 40% (even higher in children). Resistance to erythromycin was also high in S. pyogenes isolates: mean regional 33.2%, beta-lactamase-producing H. influenzae were 20%, whereas 4.4% had a beta-lactamase-negative, ampicillin-resistant phenotype. We highlight the importance of different geographical frequencies of coresistance (associations of resistance to different drugs within the same species) and coupled resistance (association of resistance between different species) probably resulting from different local coselective events.

Survey of emm-like gene sequences from pharyngeal isolates of group C and group G streptococci collected in Spain.

We conducted a nationwide surveillance of the variable 5' emm-like (M-like protein gene) sequences from 214 pharyngeal group C and group G streptococci. Almost 75% of the isolates exhibited emm or emm-like sequences previously described. We identified six new 5' emm-like regions, and almost 23% of the isolates were nontypeable. Five emm-like sequences accounted for more than 50% of the isolates in group C and group G, suggesting horizontal gene transfer between strains of different species.

Streptococcus pyogenes pharyngeal isolates with reduced susceptibility to ciprofloxacin in Spain: mechanisms of resistance and clonal diversity.

A survey of emm gene sequences and an analysis of the pulsed-field electrophoretic profiles of 30 Streptococcus pyogenes isolates with reduced susceptibilities to ciprofloxacin detected the prevalence of isolates with emm type 6 and considerable genetic diversity among isolates. The mechanism of ciprofloxacin resistance in these isolates was based on point mutations in topoisomerase IV subunit C encoded by parC, mainly replacement of serine-79 by alanine.

Effect of social and climatological factors on antimicrobial use and Streptococcus pneumoniae resistance in different provinces in Spain.

OBJECTIVES: To investigate the association between geographical differences in antibiotic consumption and resistance of Streptococcus pneumoniae to penicillin and erythromycin in 15 provinces of Spain, taking into account the potential influence of a series of social and climatological factors. METHODS: Possible correlations between prevalence of resistance to penicillin and erythromycin of S. pneumoniae, as determined in the national reference laboratory, and antibiotic consumption, and socio-economic and climatological variables were investigated. Partial correlations and multivariate linear regression were performed to assess the relative importance of variables predicting resistance and to investigate explicative factors for antibiotic consumption, respectively. RESULTS: A correlation was found between resistance and educational level, the proportion of young people in the population and climate, but was explained by their effects on differences in antibiotic use, which appeared to be the basic and only force behind resistance patterns in different geographical areas. Antibiotic use was found to be determined by the interplay of adult illiteracy, rainfall and GDP per capita. CONCLUSIONS: Interventions aimed at improving educational level and economic growth might therefore be followed by a noticeable reduction in overall antibiotic consumption, which might in turn be followed by a reduction in penicillin and erythromycin resistance in clinical isolates of S. pneumoniae.