[Epidemiology of leptospirosis in the wetlands of Southern Spain]. / Epidemiología de la leptospirosis en los humedales del sur de España.

OBJECTIVE: To find out whether the leptospirosis incidence rate among red swamp crayfish collectors in the harvesting season is higher than in the general population, and to identify risk factors and assess the direct and indirect health costs associated with leptospirosis seroconversion. METHOD: This study was carried out between 1 July 2017 and 31 March 2018 in the municipality of Isla Mayor (Seville, Spain). It took the form of a prospective cohort study (exposed population: swamp crayfish collectors; non-exposed population: general population). The population was invited to take part in a prevalence study to be conducted using the ELISA qualitative technique, and informed consent was obtained from those who agreed. Negative serology cases were then included in the cohort study. Both cohorts were monitored clinically and symptomatic cases were serology tested. A second serum sample was taken from the swamp crayfish collectors at the end of the monitoring period to detect asymptomatic cases. Serovars were confirmed by microscopic agglutination testing. A bivariate descriptive analysis was carried out and cumulative incidence and relative risk were calculated, with positive serology being taken as the dependent variable. RESULTS: A total of 278 people were included in the study, of whom 92 made up the swamp crayfish collectors cohort and 186 the general population cohort. Women made up 46.8% of the sample, but only 29.3% of the collectors cohort. The mean age was 45.1 (±16.4) years. Nine cases of seroconversion were detected: eight among swamp crayfish collectors and one in the general population. Overall cumulative incidence was therefore 3.2%: 8.7% in the exposed group and 0.5% in the non-exposed group. Relative risk was 16.2% (95% confidence interval: 2.1-127.4). The total cost of medical assistance and illness-related losses associated with leptospirosis was 1568€/case. CONCLUSIONS: Leptospirosis in Isla Mayor is strongly associated with red swamp crayfish collecting. It's incidence here is much higher than that reported in studies published in other countries.

Epidemiología de la leptospirosis en los humedales del sur de España / Epidemiology of leptospirosis in the wetlands of Southern Spain

Objetivo: Conocer si la incidencia de leptospirosis en los capturadores de cangrejo rojo durante el periodo de captura es superior a la de la población general, así como identificar factores de riesgo y estimar los costes sanitarios directos e indirectos asociados a los casos de seroconversión de dicha enfermedad.Método: Este estudio se realizó desde el 1 de julio de 2017 hasta el 31 de marzo de 2018. Se llevó a cabo un estudio de cohortes prospectivo (población expuesta: capturadores de cangrejo rojo; población no expuesta: población general) en el municipio de Isla Mayor (Sevilla, España). Previo consentimiento informado, se invitó a la población al estudio de prevalencia mediante la técnica cualitativa ELISA. Los que tuvieron serología negativa fueron incluidos en el estudio de cohortes. Ambas cohortes se siguieron clínicamente y a los casos sintomáticos se les realizó serología. A los capturadores de cangrejo rojo se les tomó una segunda muestra de suero al final del seguimiento para detectar asintomáticos. La serovariedad se confirmó mediante aglutinación microscópica. Se realizó un análisis descriptivo bivariado y se calcularon la incidencia acumulada y el riesgo relativo. La serología positiva se tomó como variable dependiente.Resultados:Se incluyeron en el estudio 278 personas, de las que 92 constituían la cohorte de capturadores y 186 la de población general. El 46,8% de la muestra eran mujeres, aunque entre los capturadores de cangrejo rojo estas solo representaban el 29,3%. La edad media de la muestra fue de 45,1 (± 16,4) años. Se detectaron nueve seroconversiones: ocho en capturadores de cangrejo rojo y una en población general. Por lo tanto, la incidencia acumulada fue de 8,7% en capturadores de cangrejo rojo y de 0,5% en población general, siendo el riesgo relativo de 16,2 (intervalo de confianza del 95%: 2,1-127,4). El coste total de la asistencia sanitaria y de las pérdidas por enfermedad asociadas a la leptospirosis fue de 1568 € por caso(AU)

Objective: To find out whether the leptospirosis incidence rate among red swamp crayfish collectors in the harvesting season is higher than in the general population, and to identify risk factors and assess the direct and indirect health costs associated with leptospirosis seroconversion. Method: This study was carried out between 1 July 2017 and 31 March 2018 in the municipality of Isla Mayor (Seville, Spain). It took the form of a prospective cohort study (exposed population: swamp crayfish collectors; non-exposed population: general population). The population was invited to take part in a prevalence study to be conducted using the ELISA qualitative technique, and informed consent was obtained from those who agreed. Negative serology cases were then included in the cohort study. Both cohorts were monitored clinically and symptomatic cases were serology tested. A second serum sample was taken from the swamp crayfish collectors at the end of the monitoring period to detect asymptomatic cases. Serovars were confirmed by microscopic agglutination testing. A bivariate descriptive analysis was carried out and cumulative incidence and relative risk were calculated, with positive serology being taken as the dependent variable. Results: A total of 278 people were included in the study, of whom 92 made up the swamp crayfish collectors cohort and 186 the general population cohort. Women made up 46.8% of the sample, but only 29.3% of the collectors cohort. The mean age was 45.1 (±16.4) years. Nine cases of seroconversion were detected: eight among swamp crayfish collectors and one in the general population. Overall cumulative incidence was therefore 3.2%: 8.7% in the exposed group and 0.5% in the non-exposed group. Relative risk was 16.2% (95% confidence interval: 2.1-127.4). The total cost of medical assistance and illness-related losses associated with leptospirosis was 1568€/case. (AU)

Brote por virus respiratorio sincitial en la Unidad de Neonatología de un hospital de tercer nivel / Bedside ultrasound in the critically ill paediatric patient

INTRODUCCIÓN: Investigación y control de un brote por virus respiratorio sincitial (VRS) que afectó a la Unidad de Neonatología (UN) de un hospital universitario de octubre a diciembre del 2012. PACIENTES Y MÉTODOS: Estudio de cohortes de los niños ingresados en la UN. Se calculó la tasa de ataque de infección y se realizaron un análisis descriptivo de los casos y un análisis multivariante de aquellas variables que mostraron ser factores de riesgo de infección por VRS. Las medidas preventivas llevadas a cabo fueron: aislamiento de contacto de casos; formación y observación de higiene de manos; exclusividad del personal sanitario para casos, restricción de visitas; estudio de portadores de VRS y profilaxis con palivizumab. RESULTADOS: El brote tuvo 3 ondas epidémicas y un total de 20 casos, de 48 niños ingresados. La tasa de ataque global fue del 42%. De los casos, la mitad fueron niños, con una edad mediana de 36 días (p25=22, p75=58). El peso al nacimiento inferior a 1.000 g (OR=23,5; p = 0,002) y tener otra infección nosocomial en la semana previa al diagnóstico de infección por VRS (OR=19,98; p = 0,016), fueron factores de riesgo independientes de infección por VRS. CONCLUSIONES: Se trató de un brote epidémico con un elevado número de casos, relacionado con el retraso en la notificación, el tiempo prolongado del estado de portador del VRS y los fallos en el cumplimiento de la higiene de manos, que favoreció la transmisión cruzada de la infección. Las medidas preventivas más eficaces fueron la observación directa de higiene de manos y supervisión de las medidas de aislamiento

INTRODUCTION: Investigation and control of a respiratory syncytial virus (RSV) outbreak that affected the Neonatal Intensive Care Unit (NICU) of a university hospital from October to December 2012. PATIENTS AND METHODS: Cohort study of children admitted to the NICU. The infection attack rate was calculated. A descriptive analysis of the cases and a multivariate analysis was performed using the variables that were shown to be risk factors for RSV infection. Preventive measures taken were: contact isolation; hand hygiene training and observation; exclusivity of a health team of nurses and physicians for positive cases, restrictions on visitor numbers; surveillance RSV testing, and palivizumab prophylaxis. RESULTS: The outbreak had three epidemic waves and 20 positive cases out of a total of 48 children admitted. The overall attack rate was 42%. Half of positive cases were children, with a median age of 36 days (p25=22, p75=58). The independent risk factors for RSV infection were birth weight below 1000 grams (OR=23.5; P=.002) and to have another nosocomial infection the week before the diagnosis of RSV infection (OR=19.98; P=.016). CONCLUSIONS: It was an outbreak with a high number of cases, due to the delay in notification, prolonged RSV carrier status, and low adherence to hand hygiene practice, which favoured the cross-transmission of infection. The most effective preventive measures were direct observation of hand hygiene and supervision of isolation measures

Influencia de la correcta identificación en la interpretación de las pruebas de sensibilidad en aislados de Aeromonas spp. productoras de bacteriemia / The relevance of correct identification and interpretation of susceptibility testing of Aeromonasspp. bacteremia isolates

OBJETIVO: Estudiar la importancia de la correcta identificación a nivel de especie así como la interpretación de las pruebas de sensibilidad en aislados de Aeromonas spp. productoras de bacteriemia mediante los métodos convencionales rutinarios y los nuevos métodos moleculares. MATERIAL Y MÉTODOS: El estudio incluyó a 22 pacientes con bacteriemia por Aeromonas hydrophila grupo, identificadas mediante el sistema MicroScan. La identificación posterior a nivel de especie se realizó por espectrometría de masas y se confirmó mediante la secuenciación del gen rpoB. La actividad de imipenem, cefotaxima, piperacilina/tazobactam, ciprofloxacino y cotrimoxazol se estudió por microdilución comercial y tiras de gradiente de antibiótico con bajo y alto inóculo. La detección de carbapenemasas se realizó mediante el test de Hodge modificado y su confirmación mediante la detección por PCR del gen cphA. RESULTADOS: Se identificaron 9 (40,9%) aislamientos como Aeromonas hydrophila, 8 (36,4%) como Aeromonas veronii y los 5 (22,7%) restantes como Aeromonas caviae. La resistencia a los antibióticos betalactámicos mediante microdilución comercial y tiras de gradiente de CMI fue, respectivamente, del 36-50% para imipenem; del 4-56% para cefotaxima; y de 27-56% para piperacilina/tazobactam. La concordancia entre el sistema automatizado y el sistema de difusión con tira de gradiente antibiótico fue, globalmente para las 3 especies, del 68% para imipenem, del 50% para cefotaxima y del 46% para piperacilina/tazobactam. No se detectó resistencia a cotrimoxazol y ciprofloxacino por ambos métodos, aunque el 22,7% de las cepas fueron resistentes a ácido nalidíxico. CONCLUSIONES: Es fundamental la identificación a nivel de especie de los aislamientos de Aeromonas spp. ya que la resistencia a betalactámicos es especie y método dependiente. Los altos porcentajes de resistencia antibiótica encontrados no aconsejan el uso de antibióticos betalactámicos y quinolonas como tratamiento empírico de la infección invasiva por Aeromonas ssp

OBJECTIVE: To assess the relevance of correct identification and interpretation of susceptibility testing of Aeromonas spp. bacteremia isolates using newly developed molecular methods in comparison to previous conventional methods. MATERIAL AND METHODS: The study included 22 patients with bacteremia due to Aeromonas hydrophila group, microbiologically characterized using the MicroScan system. Further identification to species level was performed by mass spectrometry, and confirmed by sequencing the rpoB gene. The MIC of imipenem, cefotaxime, piperacillin-tazobactam, ciprofloxacin and cotrimoxazole was studied using a commercial broth microdilution and antibiotic gradient strips with low and high inocula. Detection of carbapenemase production was performed using the modified Hodge test, and was confirmed by amplifying the cphA gene by PCR. RESULTS: A total of 9 (40.9%) isolates were identified as Aeromonas hydrophila, 8 (36.4%) as Aeromonas veronii, and the remaining 5 (22.7%) isolates as Aeromonas caviae. Resistance to beta-lactams according to both the commercial microdilution and MIC gradient strips methods was: 36%-50% to imipenem; 4%-56% to cefotaxime, and 27%-56% to piperacillin/tazobactam. The agreement between results generated by the automated system and the diffusion antibiotic gradient strip was, for all 3 species, 68% for imipenem, 50% to cefotaxime, and 46% to piperacillin/tazobactam. No resistance to cotrimoxazole and ciprofloxacin was found by either of the two methods, although 22.7% of the strains were resistant to nalidixic acid. CONCLUSIONS: It is essential to identify the isolates of Aeromonas spp. at the species level, due to the fact that beta-lactam resistance is species- and method-dependent. The high rate of resistance to beta-lactam and quinolones reduce their application as empiric treatments for invasive infection by Aeromonas ssp

[Respiratory syncytial virus outbreak in a tertiary hospital Neonatal Intensive Care Unit]. / Brote por virus respiratorio sincitial en la Unidad de Neonatología de un hospital de tercer nivel.

INTRODUCTION: Investigation and control of a respiratory syncytial virus (RSV) outbreak that affected the Neonatal Intensive Care Unit (NICU) of a university hospital from October to December 2012. PATIENTS AND METHODS: Cohort study of children admitted to the NICU. The infection attack rate was calculated. A descriptive analysis of the cases and a multivariate analysis was performed using the variables that were shown to be risk factors for RSV infection. Preventive measures taken were: contact isolation; hand hygiene training and observation; exclusivity of a health team of nurses and physicians for positive cases, restrictions on visitor numbers; surveillance RSV testing, and palivizumab prophylaxis. RESULTS: The outbreak had three epidemic waves and 20 positive cases out of a total of 48 children admitted. The overall attack rate was 42%. Half of positive cases were children, with a median age of 36 days (p25=22, p75=58). The independent risk factors for RSV infection were birth weight below 1000 grams (OR=23.5; P=.002) and to have another nosocomial infection the week before the diagnosis of RSV infection (OR=19.98; P=.016). CONCLUSIONS: It was an outbreak with a high number of cases, due to the delay in notification, prolonged RSV carrier status, and low adherence to hand hygiene practice, which favoured the cross-transmission of infection. The most effective preventive measures were direct observation of hand hygiene and supervision of isolation measures.

[The relevance of correct identification and interpretation of susceptibility testing of Aeromonas spp. bacteremia isolates]. / Influencia de la correcta identificación en la interpretación de las pruebas de sensibilidad en aislados de Aeromonas spp. productoras de bacteriemia.

OBJECTIVE: To assess the relevance of correct identification and interpretation of susceptibility testing of Aeromonas spp. bacteremia isolates using newly developed molecular methods in comparison to previous conventional methods. MATERIAL AND METHODS: The study included 22 patients with bacteremia due to Aeromonas hydrophila group, microbiologically characterized using the MicroScan system. Further identification to species level was performed by mass spectrometry, and confirmed by sequencing the rpoB gene. The MIC of imipenem, cefotaxime, piperacillin-tazobactam, ciprofloxacin and cotrimoxazole was studied using a commercial broth microdilution and antibiotic gradient strips with low and high inocula. Detection of carbapenemase production was performed using the modified Hodge test, and was confirmed by amplifying the cphA gene by PCR. RESULTS: A total of 9 (40.9%) isolates were identified as Aeromonas hydrophila, 8 (36.4%) as Aeromonas veronii, and the remaining 5 (22.7%) isolates as Aeromonas caviae. Resistance to beta-lactams according to both the commercial microdilution and MIC gradient strips methods was: 36%-50% to imipenem; 4%-56% to cefotaxime, and 27%-56% to piperacillin/tazobactam. The agreement between results generated by the automated system and the diffusion antibiotic gradient strip was, for all 3 species, 68% for imipenem, 50% to cefotaxime, and 46% to piperacillin/tazobactam. No resistance to cotrimoxazole and ciprofloxacin was found by either of the two methods, although 22.7% of the strains were resistant to nalidixic acid. CONCLUSIONS: It is essential to identify the isolates of Aeromonas spp. at the species level, due to the fact that beta-lactam resistance is species- and method-dependent. The high rate of resistance to beta-lactam and quinolones reduce their application as empiric treatments for invasive infection by Aeromonas ssp.

Molecular epidemiology and antimicrobial susceptibility of Campylobacter coliclinical isolates

INTRODUCTION: Campylobacter spp. is a major cause of acute bacterial diarrhea in humans worldwide, and C. coli is responsible for 10% of the cases. Materials and methods A study was made of the antimicrobial susceptibility using the E-test®, and the clonal relationship using PCR-RFLP, of the flaA gene, as well as PFGE techniques on 43 C. coli clinical isolates. Results Only 49% and 2% of the isolates were susceptible to erythromycin and ciprofloxacin, respectively. Imipenem and clindamyicn, with 100% and 84% of the strains, respectively, being susceptible, were the most active antimicrobials. The PCR-RFLP of flaA gene technique grouped fourteen isolates into six clusters, while the PFGE technique grouped eleven isolates into five clusters. Conclusion Ciprofloxacin and erythromycin are not suitable for the treatment of C. coli infections. Clindamycin could be considered as a therapeutic alternative in cases of enteritis, while imipenem is the best alternative for extra-intestinal infections. Both PFGE and PCR-RFLP can be useful to detect clones

INTRODUCCIÓN: Campylobacter spp. es la principal causa de diarrea bacteriana en el mundo, siendo. C. coliresponsable del 10% de los casos. MATERIAL Y MÉTODOS: Hemos estudiado la sensibilidad antibiótica mediante E-test® y la relación clonal por PCR-RFLP del gen flaA y ECP de 43 cepas de C. coli. RESULTADOS: Solo el 49 y el 2% de todas las cepas estudiadas fueron sensibles a eritromicina y a ciprofloxa cino, respectivamente. El imipenem, con el 100% de cepas sensibles, y la clindamicina, con el 84%, fueron los antibióticos más activos. La técnica de PCR-RFLP del gen flaA agrupó13 cepas en 6 clones, y la ECP,11 cepas en 5 clones. CONCLUSIONES: Ciprofloxacino y eritromicina no son antimicrobianos adecuados para tratar las infecciones por C. coli. La clindamicina se puede considerar como una alternativa terapéutica en caso de enteritis, mientras que el imipenem es la mejor alternativa en la infección extraintestinal. Tanto la ECP como la PCR-RFLP del gen flaA son técnicas útiles para detectar clones

Molecular epidemiology and antimicrobial susceptibility of Campylobacter coli clinical isolates.

INTRODUCTION: Campylobacter spp. is a major cause of acute bacterial diarrhea in humans worldwide, and C. coli is responsible for 10% of the cases. MATERIALS AND METHODS: A study was made of the antimicrobial susceptibility using the E-test(®), and the clonal relationship using PCR-RFLP, of the flaA gene, as well as PFGE techniques on 43 C. coli clinical isolates. RESULTS: Only 49% and 2% of the isolates were susceptible to erythromycin and ciprofloxacin, respectively. Imipenem and clindamyicn, with 100% and 84% of the strains, respectively, being susceptible, were the most active antimicrobials. The PCR-RFLP of flaA gene technique grouped fourteen isolates into six clusters, while the PFGE technique grouped eleven isolates into five clusters. CONCLUSION: Ciprofloxacin and erythromycin are not suitable for the treatment of C. coli infections. Clindamycin could be considered as a therapeutic alternative in cases of enteritis, while imipenem is the best alternative for extra-intestinal infections. Both PFGE and PCR-RFLP can be useful to detect clones.

Genetic variants of the MBL2 gene are associated with mortality in pneumococcal sepsis.

Studies evaluating associations between polymorphisms of innate immunity genes and prognosis of infectious diseases have yielded conflicting results. Our aim was to assess the impact on mortality of different genotypic variants of the innate immunity in patients with pneumococcal sepsis. All adults admitted to the hospital with diagnosis of sepsis caused by Streptococcus pneumoniae were enrolled and single-nucleotide polymorphisms (SNP) in mannose-binding lectin 2 (MBL2), toll-like receptor (TLR) 2, TLR4, and Fcγ receptor IIa genes were genotyped. Underlying diseases, severity of illness, and antibiotic management were also recorded. We included 117 patients: 98 pneumonias (83.6%), 17 meningitis (14.5%), and 2 patients (1.9%) with primary pneumococcal bacteremia. Allelic variants of the MBL2 gene (individuals heterozygous or homozygous for one of the 3 allelic variants B, C, and D: AO/OO) were present in 37 patients (32%), T399I polymorphism in TLR4 in 19 (16.2%), TLR4 D299G/T399I in 11 (9.4%), TLR2 R753Q in 3 (2.5%), and FcγRIIa-R/R131 in 26 patients (23%). Factors associated independently with in-hospital mortality were SNP MBL2 AO/OO (adjusted hazard ratios [aHR] 3.2, 95% confidence interval [CI] 1.01-9.8) and septic shock (aHR 15.3, 95% CI 3.5-36.5), whereas first adequate antibiotic dose ≤ 4 h was a protective factor (aHR 0.2, 95% CI 0.06-0.8). SNP MBL2 AO/OO (aHR 2.2, 95% CI 1.1-8.1) remained as a variable independently associated with 90-day mortality. In conclusion, variant alleles in the MBL2 gene are independently associated with in-hospital and medium-term mortalities in patients admitted to the hospital with pneumococcal sepsis.

Risk factors for fluconazole-resistant candidemia.

Previous studies have sought to determine the risk factors associated with candidemia caused by non-albicans Candida spp. or with potentially fluconazole-resistant Candida spp. (C. glabrata and C. krusei). Non-albicans Candida strains are a heterogeneous group that includes species with different levels of virulence, and only a limited number of C. glabrata isolates are resistant to fluconazole. We set out to identify the risk factors associated with microbiologically proven fluconazole-resistant candidemia. A prospective study including adult patients with candidemia was performed. Data were collected on patient demographics; underlying diseases; exposure to corticosteroids, antibiotics, or fluconazole; and invasive procedures. Risk factors associated either with non-albicans Candida spp. or potentially fluconazole-resistant Candida spp. (C. glabrata or C. krusei) or with Candida spp. with microbiologically confirmed fluconazole resistance were assessed using logistic regressions. We included 226 candidemia episodes. Non-albicans Candida isolates accounted for 53.1% of the fungal isolates, but only 18.2% of the cases were caused by potentially fluconazole-resistant organisms. Thirty isolates exhibited microbiologically confirmed fluconazole resistance. The multivariate analysis revealed that independent predictors associated with fluconazole-resistant Candida spp. were neutropenia (odds ratio [OR]=4.94; 95% confidence interval [CI]=1.50 to 16.20; P=0.008), chronic renal disease (OR=4.82; 95% CI=1.47 to 15.88; P=0.01), and previous fluconazole exposure (OR=5.09; 95% CI=1.66 to 15.6; P=0.004). Independently significant variables associated with non-albicans Candida bloodstream infection or with potentially fluconazole-resistant Candida spp. did not include previous fluconazole exposure. We concluded that prior fluconazole treatment is an independent risk factor only for candidemia caused by microbiologically confirmed fluconazole resistant species. Our findings may be of value for selecting empirical antifungal therapy.

Determinants of outcome in patients with bacteraemic pneumococcal pneumonia: importance of early adequate treatment.

We set out to determine the factors influencing mortality in 125 adult patients with bacteraemic pneumococcal community-acquired pneumonia (CAP), assessing the impact on outcomes of early adequate therapy in particular. Presumed prognostic factors with p < 0.1 in the unadjusted model were subjected to multivariate Cox regression analysis, with in-hospital and 90-day mortalities as the dependent variables. A time period of >4 h from admission to start of adequate antibiotic treatment (adjusted hazard ratio (aHR) 2.62, 95% confidence interval (CI) 1.06-6.45; p =0.037) and severe sepsis or septic shock (aHR 5.06, 95% CI 1.63-15.71; p = 0.005) were independently associated with in-hospital mortality. Variables associated with 90-day mortality were Charlson comorbidity index (aHR 1.17, 95% CI 1.02-1.34; p = 0.018), severe sepsis or septic shock (aHR 3.03, 95% CI 1.22-7.51; p = 0.016) and delay of adequate antibiotic therapy >4 h (aHR 2.21, 95% CI 1.01-4.86; p = 0.048). The use of combination therapy was not included in these models but was a protective factor for delayed adequate therapy (aHR 0.53, 95% CI 0.29-0.95; p = 0.033). Administration of adequate antimicrobial therapy within 4 h of arrival is a critical determinant of survival in patients with bacteraemic pneumococcal CAP.

Antibióticos y resistencia antibiótica en la quimioprofilaxis de la enfermedad meningocócica / Antibiotics and antibiotic resistance in chemoprophylaxis of meningococcal disease

La profilaxis antibiótica debe considerarse en personas con contacto con algún caso de enfermedad meningocócica o en poblaciones con altos porcentajes de portadores de N. meningitidis. El uso de antibióticos produce una reducción significativa del riesgo de enfermedad entre los contactos; así, la rifampicina, la ciprofloxacina y la ceftriaxona se consideran como las mejores opciones para la quimioprofilaxis, sin embargo su empleo está asociado con el aumento de resistencia antibiótica. Actualmente, la tendencia a la aparición de meningococos resistentes a la rifampicina después de la profilaxis es un aspecto reconocido, aunque parece que no es un fenómeno ampliamente extendido. La aparición de resistencia de alto nivel a la rifampicina está provocada por mutaciones en el gen rpoB, aunque se puede asociar a mutaciones del locus mtr implicadas en mecanismos de expulsión y bombeo. Sin embargo, los cambios en el gen rpoB dan lugar a cepas poco adaptadas a la supervivencia y este costo biológico podría explicar la ausencia de diseminación clonal de los aislamientos con resistencia adquirida a la rifampicina. La resistencia o sensibilidad disminuida a la ciprofloxacina se relaciona con mutaciones en la región determinante de resistencia a quinolonas (QRDR) del gen gyrA, además existen datos que apoyan la existencia de mecanismos de expulsión. Hasta el momento, la resistencia o la sensibilidad reducida a las quinolonas ha avanzado lentamente. No se han comunicado problemas de resistencia a la ceftriaxona, siendo la opción más segura para su uso en quimioprofilaxis.La espiramicina no es una opción adecuada aunque sigue siendo recomendada por la OMS.

[Update on the epidemiology, diagnosis and treatment of sexually-transmitted infections]. / Panorama actual de la epidemiología, diagnóstico y tratamiento de las infecciones de transmisión sexual.

In the last decade, cases of sexually-transmitted infections (STIs) have progressively increased in Europe. The reasons for this increase are unclear, but may involve changes in social behavior, migration and international travel, coupled with the emergence of risk groups that have not been taken into sufficient consideration to date. The routine use of molecular diagnostic techniques for many of these infections has solved many problems of sensitivity and the suitability of samples for microbiological diagnosis: non-invasive samples can be used, which has undoubtedly contributed to the increase in the number of cases. Moreover, molecular methods have also been introduced for antibiotic and antiviral susceptibility testing, as well as for molecular characterization of clinical isolates. All of these factors, together with the approval of the vaccine against the human papillomavirus, have changed the landscape of STIs across Europe.

Panorama actual de la epidemiología, diagnóstico y tratamiento de las infecciones de transmisión sexual / Update on the epidemiology, diagnosis and treatment of sexually-transmitted infections

En la última década se ha constatado un aumentoprogresivo de los casos de infecciones de transmisiónsexual en el territorio europeo. Las causas de esteaumento no están claras, pero parece influido por cambiosen las conductas sociales, los fenómenos migratorios y losviajes internacionales, junto con la aparición de grupos deriesgo no suficientemente valorados hasta ahora. Lautilización habitual de técnicas de diagnóstico molecularpara muchas de estas infecciones ha resuelto muchosproblemas de sensibilidad e idoneidad de las muestraspara el diagnóstico microbiológico, pudiéndose emplearmuestras no invasivas, y ha contribuido, sin duda, a esteaumento de casos. Por otro lado, los métodos molecularestambién se van implantando en el estudio de lasensibilidad a los antibióticos y antivirales, así como parala caracterización molecular de los aislados. Todo loanterior, junto a la aprobación de la vacuna frente al virusdel papiloma humano, ha cambiado el panorama de lasinfecciones de transmisión sexual en el territorio europeo(AU)

In the last decade, cases of sexually-transmitted infections(STIs) have progressively increased in Europe. The reasonsfor this increase are unclear, but may involve changes insocial behavior, migration and international travel, coupledwith the emergence of risk groups that have not beentaken into sufficient consideration to date.The routine use of molecular diagnostic techniques formany of these infections has solved many problems of sensitivity and the suitability of samples formicrobiological diagnosis: non-invasive samples can beused, which has undoubtedly contributed to the increasein the number of cases. Moreover, molecular methodshave also been introduced for antibiotic and antiviralsusceptibility testing, as well as for molecularcharacterization of clinical isolates. All of these factors,together with the approval of the vaccine against thehuman papillomavirus, have changed the landscape ofSTIs across Europe(AU)

Bovine tuberculosis in Doñana Biosphere Reserve: the role of wild ungulates as disease reservoirs in the last Iberian lynx strongholds.

Doñana National Park (DNP) in southern Spain is a UNESCO Biosphere Reserve where commercial hunting and wildlife artificial feeding do not take place and traditional cattle husbandry still exists. Herein, we hypothesized that Mycobacterium bovis infection prevalence in wild ungulates will depend on host ecology and that variation in prevalence will reflect variation in the interaction between hosts and environmental risk factors. Cattle bTB reactor rates increased in DNP despite compulsory testing and culling of infected animals. In this study, 124 European wild boar, 95 red deer, and 97 fallow deer were sampled from April 2006 to April 2007 and analyzed for M. bovis infection. Modelling and GIS were used to identify risk factors and intra and inter-species relationships. Infection with M. bovis was confirmed in 65 (52.4%) wild boar, 26 (27.4%) red deer and 18 (18.5%) fallow deer. In the absence of cattle, wild boar M. bovis prevalence reached 92.3% in the northern third of DNP. Wild boar showed more than twice prevalence than that in deer (p<0.001). Modelling revealed that M. bovis prevalence decreased from North to South in wild boar (p<0.001) and red deer (p<0.01), whereas no spatial pattern was evidenced for fallow deer. Infection risk in wild boar was dependent on wild boar M. bovis prevalence in the buffer area containing interacting individuals (p<0.01). The prevalence recorded in this study is among the highest reported in wildlife. Remarkably, this high prevalence occurs in the absence of wildlife artificial feeding, suggesting that a feeding ban alone would have a limited effect on wildlife M. bovis prevalence. In DNP, M. bovis transmission may occur predominantly at the intra-species level due to ecological, behavioural and epidemiological factors. The results of this study allow inferring conclusions on epidemiological bTB risk factors in Mediterranean habitats that are not managed for hunting purposes. Our results support the need to consider wildlife species for the control of bTB in cattle and strongly suggest that bTB may affect animal welfare and conservation.

Utilidad de las técnicas de biología molecular en el diagnóstico de las infecciones de transmisión sexual y otras infecciones genitales / Utility of molecular biology techniques in the diagnosis of sexually transmitted diseases and genital infections

Históricamente, el diagnóstico de las infecciones de transmisión sexual ha sido difícil. La introducción en el diagnóstico microbiológico de las técnicas de biología molecular y su aplicación a muestras no invasivas ha permitido importantes avances en su diagnóstico. En general, la detección de Neisseria gonorrhoeae mediante técnicas de biología molecular proporciona un diagnóstico presuntivo y requiere confirmación por cultivo en zonas de baja prevalencia. Para Chlamydia trachomatis, estas técnicas se consideran como las más sensibles y específicas, tanto para estudios de cribado poblacional, como para el diagnóstico de pacientes sintomáticos. El diagnóstico de Mycoplasma genitalium por cultivo es muy lento, por ello, las técnicas moleculares son las únicas que pueden aportar información diagnóstica relevante. Para Treponema pallidum, las técnicas moleculares pueden aportar ventajas en el diagnóstico directo de la infección. Respecto a la donovaniosis, las técnicas moleculares no están establecidas para el diagnóstico sistemático, aunque se recomiendan en manos expertas. En el caso de Haemophilus ducreyi, las dificultades del cultivo y su baja sensibilidad aconsejan el uso de métodos moleculares. En el herpes genital, las técnicas moleculares han comenzado a recomendarse para el diagnóstico sistemático y pueden convertirse en la técnica de referencia en poco tiempo. Para otras infecciones genitales, como vaginosis bacteriana, vulvovaginitis candidiásica y tricomoniasis, los métodos moleculares para el diagnóstico están poco establecidos. Respecto a las verrugas genitales, las técnicas de cribado y genotipado disponibles para muestras endocervicales podrían utilizarse para ciertas poblaciones, aunque no se han validado para este cometido(AU)

Historically, the diagnosis of sexually transmitted diseases (STDs) has been difficult. The introduction of molecular biology techniques in microbiological diagnosis and their application to non-invasive samples has produced significant advances in the diagnosis of these diseases. Overall, detection of Neisseria gonorrhoeae by molecular biology techniques provides a presumptive diagnosis and requires confirmation by culture in areas with a low prevalence. For Chlamydia trachomatis infections, these techniques are considered to be the most sensitive and specific procedures for mass screening studies, as well as for the diagnosis of symptomatic patients. Diagnosis of Mycoplasma genitalium infection by culture is very slow and consequently molecular techniques are the only procedures that can provide relevant diagnostic information. For Treponema pallidum, molecular techniques can provide direct benefits in the diagnosis of infection. Molecular techniques are not established for the routine diagnosis of donovanosis, but can be recommended when performed by experts. Molecular methods are advisable in Haemophilus ducreyi, because of the difficulties of culture and its low sensitivity. In genital herpes, molecular techniques have begun to be recommended for routine diagnosis and could soon become the technique of choice. For other genital infections, bacterial vaginosis, vulvovaginal candidosis and trichomoniasis, diagnosis by molecular methods is poorly established. With genital warts, techniques available for screening and genotyping of endocervical samples could be used for certain populations, but are not validated for this purpose(AU)

[Utility of molecular biology techniques in the diagnosis of sexually transmitted diseases and genital infections]. / Utilidad de las técnicas de biología molecular en el diagnóstico de las infecciones de transmisión sexual y otras infecciones genitales.

Historically, the diagnosis of sexually transmitted diseases (STDs) has been difficult. The introduction of molecular biology techniques in microbiological diagnosis and their application to non-invasive samples has produced significant advances in the diagnosis of these diseases. Overall, detection of Neisseria gonorrhoeae by molecular biology techniques provides a presumptive diagnosis and requires confirmation by culture in areas with a low prevalence. For Chlamydia trachomatis infections, these techniques are considered to be the most sensitive and specific procedures for mass screening studies, as well as for the diagnosis of symptomatic patients. Diagnosis of Mycoplasma genitalium infection by culture is very slow and consequently molecular techniques are the only procedures that can provide relevant diagnostic information. For Treponema pallidum, molecular techniques can provide direct benefits in the diagnosis of infection. Molecular techniques are not established for the routine diagnosis of donovanosis, but can be recommended when performed by experts. Molecular methods are advisable in Haemophilus ducreyi, because of the difficulties of culture and its low sensitivity. In genital herpes, molecular techniques have begun to be recommended for routine diagnosis and could soon become the technique of choice. For other genital infections, bacterial vaginosis, vulvovaginal candidosis and trichomoniasis, diagnosis by molecular methods is poorly established. With genital warts, techniques available for screening and genotyping of endocervical samples could be used for certain populations, but are not validated for this purpose.

[Detection of inducible resistance to clindamycin in cutaneous isolates of Staphylococcus spp. by phenotypic and genotypic methods]. / Detección de resistencia inducible a clindamicina en aislados cutáneos de Staphylococcus spp. por métodos fenotípicos y genotípicos.

INTRODUCTION: Resistance to macrolides, lincosamides and type B streptogramins (MLSB) in Staphylococcus isolates can be due to several mechanisms. The most important are an active efflux mechanism (MSB phenotype) and ribosomal target modification (MLSB phenotype); this latter mechanism confers resistance to all three groups of antimicrobials (MLSB resistance). Expression of MLSB resistance can be constitutive (cMLSB) or inducible (iMLSB). METHODS: A group of 117 erythromycin-resistant Staphylococcus spp. clinical isolates from cutaneous samples were selected from 536 recent clinical isolates of this microorganism. Resistance phenotypes were determined by the double disk diffusion test. Presence of the ermA, ermC, ermB and msrA genes was detected by real time PCR. RESULTS: The MSB phenotype was the most common, comprising 11.2% (7.2% in S. aureus and 23% in CoNS) of the erythromycin-resistant strains. The rate of iMLSB resistance was significantly higher, 7.4% (5.2% in S. aureus and 14% in CoNS), than the rate of cMLSB resistance, 3.2% (1.7% in S. aureus and 7.4% in CoNS). The msrA gene was present in all isolates with the MSB phenotype, and the ermC gene was the most common among clindamycin-resistant strains with the MLSB phenotype (constitutive or inducible). CONCLUSION: The good correlation between the phenotypic (disk-diffusion) and genotypic (real time PCR) methods used allows prediction of the mechanisms of erythromycin and clindamycin resistance, provides insight into the epidemiological differences in their distribution, and is an aid to selecting the most appropriate antimicrobial therapy.

Detección de resistencia inducible a clindamicina en aislados cutáneos de Staphylococcus spp. por métodos fenotípicos y genotípicos / Detection of inducible resistance to clindamycin in cutaneous isolates of Staphylococcus spp. by phenotypic and genotypic methods

Introducción. La resistencia de estafilococos a macrólidos, lincosamidas y estreptograminas del tipo B (antibióticos MLSB) puede ser debida a varios mecanismos de resistencia y entre ellos los dos más importantes son la expulsión activa (fenotipo MSB) y la modificación de la diana en el ribosoma (fenotipo MLSB). Este último mecanismo confiere resistencia cruzada a los 3 grupos de antimicrobianos (resistencia MLSB). La expresión fenotípica de la resistencia MLSB puede ser de carácter constitutivo (cMLSB) o inducible (iMLSB). Métodos. Se estudiaron 117 estafilococos resistentes a eritromicina procedentes de muestras cutáneas que fueron seleccionados a partir de 536 aislados clínicos de estafilococos. El estudio fenotípico se realizó mediante la técnica de difusión por doble disco. La presencia de los genes ermA, ermC, ermB y msrA implicados en la resistencia se estudiaron por reacción en cadena de la polimerasa (PCR) en tiempo real. Resultados. El fenotipo MSB fue el más frecuente, encontrándose en el 11,2% de las cepas (7,2% Staphylococcus aureus y 23% ECN) y la tasa de resistencia inducible iMLSB, fue estadísticamente significativa más alta 7,4% (5,2% en S. aureus y 14% en ECN) que la tasa de resistencia constitutiva cMLSB, 3,2% (1,7% en S. aureus y 7,4% en ECN). Todos los aislados con el fenotipo MSB presentaron el gen msrA y el gen ermC fue el más frecuentemente detectado en las cepas resistentes a clindamicina con fenotipo MLSB (constitutivo o inducible). Conclusión. La buena correlación entre los métodos fenotípicos (doble difusión con discos) y genotípicos permiten inferir el mecanismo de resistencia a eritromicina y clindamicina, seleccionar el tratamiento antimicrobiano más adecuado, así como apreciar las diferencias epidemiológicas en su distribución (AU)

Introduction. Resistance to macrolides, lincosamides and type B streptogramins (MLSB) in Staphylococcus isolates can be due to several mechanisms. The most important are an active efflux mechanism (MSB phenotype) and ribosomal target modification (MLSB phenotype); this latter mechanism confers resistance to all three groups of antimicrobials (MLSB resistance). Expression of MLSB resistance can be constitutive (cMLSB) or inducible (iMLSB). Methods. A group of 117 erythromycin-resistant Staphylococcus spp. clinical isolates from cutaneous samples were selected from 536 recent clinical isolates of this microorganism. Resistance phenotypes were determined by the double disk diffusion test. Presence of the ermA, ermC, ermB and msrA genes was detected by real time PCR. Results. The MSB phenotype was the most common, comprising 11.2% (7.2% in S. aureus and 23% in CoNS) of the erythromycin-resistant strains. The rate of iMLSB resistance was significantly higher, 7.4% (5.2% in S. aureus and 14% in CoNS), than the rate of cMLSB resistance, 3.2% (1.7% in S. aureus and 7.4% in CoNS). The msrA gene was present in all isolates with the MSB phenotype, and the ermC gene was the most common among clindamycin-resistant strains with the MLSB phenotype (constitutive or inducible). Conclusion. The good correlation between the phenotypic (disk-diffusion) and genotypic (real time PCR) methods used allows prediction of the mechanisms of erythromycin and clindamycin resistance, provides insight into the epidemiological differences in their distribution, and is an aid to selecting the most appropriate antimicrobial therapy (AU)

[Detection of methicillin resistance and identification of Staphylococcus spp. from positive blood culture bottles using the mecA and nucA genes with the LightCycler System]. / Detección de la resistencia a meticilina e identificación de Staphylococcus spp. en hemocultivos positivos amplificando los genes mecA y nucA con el sistema LightCycler.

INTRODUCTION: The aim of this study was to evaluate the feasibility of detecting Staphylococcus aureus and coagulase-negative staphylococci (CoNS) and of identifying methicillin resistance directly in positive BACTEC blood culture bottles using the LightCycler system. METHODS: One hundred thirty-one positive blood culture bottles in which Gram-positive cocci in cluster were observed after Gram staining and 40 positive bottles with microorganisms other than staphylococci were studied. A molecular assay based on an automated DNA extraction protocol with a MagNA Pure LC instrument was used. Oligonucleotide primers and fluorescence-labeled hybridization probes were designed for amplification and sequence-specific detection of both a 408-pb fragment within the mecA gene and a 279-pb fragment within the S. aureus-specific nucA gene. RESULTS: All the bottles that yielded methicillin-resistant S. aureus (MRSA), methicillin-sensitive S. aureus (MSSA) or methicillin-resistant CoNS (MRCoNS) strains were correctly identified by the nucA and mecA PCR assays. One bottle that yielded a mixed culture of MSSA and MRCoNS gave positive results for both genes. In the 21 bottles with methicillin-susceptible CoNS (MSCoNS), nucA PCR were negative, but two of these bottles gave positive results for the mecA gene. The sensitivity and specificity of the nucA gene assay were 100%. The sensitivity and specificity of the PCR assay for detection of methicillin resistance with the mecA gene were 100% and 97.5%, respectively. CONCLUSION: This is a sensitive and highly specific method for identifying staphylococci in positive blood cultures, allowing discrimination between methicillin-susceptible and -resistant strains in less than 3 hours after Gram stain.