Global Emergence of Resistance to Fluconazole and Voriconazole in Candida parapsilosis in Tertiary Hospitals in Spain During the COVID-19 Pandemic.

Background: Candida parapsilosis is a frequent cause of candidemia worldwide. Its incidence is associated with the use of medical implants, such as central venous catheters or parenteral nutrition. This species has reduced susceptibility to echinocandins, and it is susceptible to polyenes and azoles. Multiple outbreaks caused by fluconazole-nonsusceptible strains have been reported recently. A similar trend has been observed among the C. parapsilosis isolates received in the last 2 years at the Spanish Mycology Reference Laboratory. Methods: Yeast were identified by molecular biology, and antifungal susceptibility testing was performed using the European Committee on Antimicrobial Susceptibility Testing protocol. The ERG11 gene was sequenced to identify resistance mechanisms, and strain typing was carried out by microsatellite analysis. Results: We examined the susceptibility profile of 1315 C. parapsilosis isolates available at our reference laboratory between 2000 and 2021, noticing an increase in the number of isolates with acquired resistance to fluconazole, and voriconazole has increased in at least 8 different Spanish hospitals in 2020-2021. From 121 recorded clones, 3 were identified as the most prevalent in Spain (clone 10 in Catalonia and clone 96 in Castilla-Leon and Madrid, whereas clone 67 was found in 2 geographically unrelated regions, Cantabria and the Balearic Islands). Conclusions: Our data suggest that concurrently with the coronavirus disease 2019 pandemic, a selection of fluconazole-resistant C. parapsilosis isolates has occurred in Spain, and the expansion of specific clones has been noted across centers. Further research is needed to determine the factors that underlie the successful expansion of these clones and their potential genetic relatedness.

Infección bacteriana/fúngica en pacientes con COVID-19 ingresados en un hospital de tercer nivel de Castilla y León, España / Bacterial/fungal infection in hospitalized patients with COVID-19 in a tertiary hospital in the Community of Castilla y León, Spain

IntroducciónLa coinfección/sobreinfección bacteriana/fúngica contribuye al incremento de la morbimortalidad de las infecciones respiratorias (IRs) virales. El objetivo de este trabajo es conocer la incidencia de estas infecciones en los pacientes hospitalizados por COVID-19.MétodoEstudio retrospectivo observacional de todos los pacientes ingresados por COVID-19 e infección bacteriana/fúngica en el Hospital Clínico de Valladolid (1 marzo-31 mayo, 2020). Comparación de datos demográficos, clínicos y microbiológicos en función del ingreso en UCI e identificación de los factores predictores de mortalidad mediante regresión logística multivariante.ResultadosDe 712 pacientes con COVID-19, 113 (16%) presentaron coinfección/ sobreinfección bacteriana/fúngica. Mediana de edad 73 años (RIQ 57-89), 59% de ellos hombres. Perfil del paciente de UCI (44%): hombre con neumonía por SARS-CoV-2, leucocitosis, interleucina-6 elevada, con interferón β-1b y tocilizumab y sobreinfección (p < 0,05). El 5% (39/712) de los pacientes presentaron una coinfección. Streptococcus pneumoniae (6) y Staphylococcus aureus (6) fueron los principales patógenos de las coinfecciones respiratorias (18). El 11% (80/712) se sobreinfectaron. Las infecciones más frecuentes fueron las urinarias (53) e IRs (39). Acinetobacter baumannii multirresistente fue el principal agente de la IR y la bacteriemia. Un brote por A. baumannii contribuyó a este resultado. Tres pacientes se diagnosticaron como probable aspergilosis pulmonar. La mortalidad fue superior en los pacientes de UCI (50 vs. 29%; p = 0,028). Factores predictores de mortalidad: hombre con varias comorbilidades, neumonía por SARS-CoV-2, bacteriemia y sobreinfectado por A. baumannii.ConclusiónEl brote por A. baumannii fue determinante en la incidencia de la infección y en la morbimortalidad de los pacientes de UCI.

IntroductionBacterial/fungal coinfection and superinfections contribute to the increased morbi-mortality of viral respiratory infections (RIs). The main objective of this study was to determine the incidence of these infections in hospitalized patients with COVID-19.MethodRetrospective observational study of all patients admitted for COVID-19 and bacterial/fungal infections at the Hospital Clínico Universitario of Valladolid, Spain (March 1-May 31, 2020). Demographic, clinical and microbiological data were compared based on Intensive Care Unit (ICU) admission and predictors of mortality by were identified using multivariate logistic regression analyses.ResultsOf the 712 COVID-19 patients, 113 (16%) presented bacterial/fungal coinfections or superinfections. Their median age was 73 years (IQR 57-89) and 59% were men. The profiles of ICU patients (44%) included male, SARS-CoV-2 pneumonia, leukocytosis, elevated inteleukin-6, with interferon β-1b and tocilizumab and superinfection (p < 0.05). Coinfections were diagnosed in 5% (39/712) patients. Most common pathogens of respiratory coinfection (18) were Streptococcus pneumoniae (6) and Staphylococcus aureus (6). Superinfections were detected in 11% (80/712) patients. Urinary (53) and RIs (39) constituted the majority of superinfections Acinetobacter baumannii multidrug-resistant was the main agent of IR and bacteremia. An outbreak of A. baumannii contributed to this result. Three patients were considered to have probable pulmonary aspergillosis. Mortality was higher in UCI patients (50 vs. 29%; p = 0.028). The predictive factors of mortality included being a male with various comorbidities, SARS-CoV-2 pneumonia, bacteremia and superinfections from A. baumannii.ConclusionThe outbreak of A. baumannii was a determining factor in the increases of the incidence of infection and the morbi-mortality of ICU patients.

Bacterial/fungal infection in hospitalized patients with COVID-19 in a tertiary hospital in the Community of Castilla y León, Spain.

INTRODUCTION: Bacterial/fungal coinfection and superinfections contribute to the increased morbi-mortality of viral respiratory infections (RIs). The main objective of this study was to determine the incidence of these infections in hospitalized patients with COVID-19. METHOD: Retrospective observational study of all patients admitted for COVID-19 and bacterial/fungal infections at the Hospital Clínico Universitario of Valladolid, Spain (March 1-May 31, 2020). Demographic, clinical and microbiological data were compared based on Intensive Care Unit (ICU) admission and predictors of mortality by were identified using multivariate logistic regression analyses. RESULTS: Of the 712 COVID-19 patients, 113 (16%) presented bacterial/fungal coinfections or superinfections. Their median age was 73 years (IQR 57-89) and 59% were men. The profiles of ICU patients (44%) included male, SARS-CoV-2 pneumonia, leukocytosis, elevated inteleukin-6, with interferon ß-1b and tocilizumab and superinfection (p < 0.05). Coinfections were diagnosed in 5% (39/712) patients. Most common pathogens of respiratory coinfection (18) were Streptococcus pneumoniae (6) and Staphylococcus aureus (6). Superinfections were detected in 11% (80/712) patients. Urinary (53) and RI (39) constituted the majority of superinfections Acinetobacter baumannii multidrug-resistant was the main agent of IR and bacteremia. An outbreak of A. baumannii contributed to this result. Three patients were considered to have probable pulmonary aspergillosis. Mortality was higher in UCI patients (50% vs. 29%, p = 0.028). The predictive factors of mortality included being a male with various comorbidities, SARS-CoV-2 pneumonia, bacteremia and superinfections from A. baumannii. CONCLUSION: The outbreak of A. baumannii was a determining factor in the increases of the incidence of infection and the morbi-mortality of ICU patients.

Infección bacteriana/fúngica en pacientes con COVID-19 ingresados en un hospital de tercer nivel de Castilla y León, España / Bacterial/fungal infection in hospitalized patients with COVID-19 in a tertiary hospital in the Community of Castilla y León, Spain

INTRODUCCIÓN: La coinfección/sobreinfección bacteriana/fúngica contribuye al incremento de la morbimortalidad de las infecciones respiratorias (IRs) virales. El objetivo de este trabajo es conocer la incidencia de estas infecciones en los pacientes hospitalizados por COVID-19. MÉTODO: Estudio retrospectivo observacional de todos los pacientes ingresados por COVID-19 e infección bacteriana/fúngica en el Hospital Clínico de Valladolid (1 marzo-31 mayo, 2020). Comparación de datos demográficos, clínicos y microbiológicos en función del ingreso en UCI e identificación de los factores predictores de mortalidad mediante regresión logística multivariante. RESULTADOS: De 712 pacientes con COVID-19, 113 (16%) presentaron coinfección/ sobreinfección bacteriana/fúngica. Mediana de edad 73 años (RIQ 57-89), 59% de ellos hombres. Perfil del paciente de UCI (44%): hombre con neumonía por SARS-CoV-2, leucocitosis, interleucina-6 elevada, con interferón β-1b y tocilizumab y sobreinfección (p < 0,05). El 5% (39/712) de los pacientes presentaron una coinfección. Streptococcus pneumoniae (6) y Staphylococcus aureus (6) fueron los principales patógenos de las coinfecciones respiratorias (18). El 11% (80/712) se sobreinfectaron. Las infecciones más frecuentes fueron las urinarias (53) e IRs (39). Acinetobacter baumannii multirresistente fue el principal agente de la IR y la bacteriemia. Un brote por A. baumannii contribuyó a este resultado. Tres pacientes se diagnosticaron como probable aspergilosis pulmonar. La mortalidad fue superior en los pacientes de UCI (50 vs. 29%; p = 0,028). Factores predictores de mortalidad: hombre con varias comorbilidades, neumonía por SARS-CoV-2, bacteriemia y sobreinfectado por A. baumannii. CONCLUSIÓN: El brote por A. baumannii fue determinante en la incidencia de la infección y en la morbimortalidad de los pacientes de UCI

INTRODUCTION: Bacterial/fungal coinfection and superinfections contribute to the increased morbi-mortality of viral respiratory infections (RIs). The main objective of this study was to determine the incidence of these infections in hospitalized patients with COVID-19. METHOD: Retrospective observational study of all patients admitted for COVID-19 and bacterial/fungal infections at the Hospital Clínico Universitario of Valladolid, Spain (March 1-May 31, 2020). Demographic, clinical and microbiological data were compared based on Intensive Care Unit (ICU) admission and predictors of mortality by were identified using multivariate logistic regression analyses. RESULTS: Of the 712 COVID-19 patients, 113 (16%) presented bacterial/fungal coinfections or superinfections. Their median age was 73 years (IQR 57-89) and 59% were men. The profiles of ICU patients (44%) included male, SARS-CoV-2 pneumonia, leukocytosis, elevated inteleukin-6, with interferon beta-1b and tocilizumab and superinfection (p < 0.05). Coinfections were diagnosed in 5% (39/712) patients. Most common pathogens of respiratory coinfection (18) were Streptococcus pneumoniae (6) and Staphylococcus aureus (6). Superinfections were detected in 11% (80/712) patients. Urinary (53) and RIs (39) constituted the majority of superinfections Acinetobacter baumannii multidrug-resistant was the main agent of IR and bacteremia. An outbreak of A. baumannii contributed to this result. Three patients were considered to have probable pulmonary aspergillosis. Mortality was higher in UCI patients (50 vs. 29%; p = 0.028). The predictive factors of mortality included being a male with various comorbidities, SARS-CoV-2 pneumonia, bacteremia and superinfections from A. baumannii. CONCLUSION: The outbreak of A. baumannii was a determining factor in the increases of the incidence of infection and the morbi-mortality of ICU patients

Sinovitis por Mycobacterium heckeshornense en paciente con artritis reumatoide en tratamiento con infliximab / Synovitis due to Mycobacterium heckeshornense in a patient with rheumatoid arthritis and treatment with infliximab

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A strategy based on Amplified Fragment Length Polymorphism (AFLP) for routine genotyping of nontuberculous mycobacteria at the clinical laboratory.

The increasing worldwide incidence of mycobacteriosis and the need to achieve improved clinical management makes nontuberculous mycobacteria (NTM) genotyping a useful tool. However, because of technical difficulties, medium size microbiology laboratories do not attempt to compare the genetic patterns that each of their isolates present. We have aimed to optimize a genotyping method with a reduced hands-on experimental time and that requires few technical resources. A strategy based on the Amplified Fragment Length Polymorphism (AFLP) methodology was developed using two rare-cutters enzymes (SacI and BglII). One out of seven primers was sequentially used in each amplification reaction that was analyzed by agarose gel electrophoresis. This approach makes it possible the timely genotyping of a moderate number of strains and its characterization without the need of image analysis software. We have genotyped 28 Mycobacterium intracellulare and 4 M. abscessus. Clinical researchers are encouraged to routinely genotype their NTM isolates.

Bacterial/fungal infection in hospitalized patients with COVID-19 in a tertiary hospital in the Community of Castilla y León, Spain. / Infección bacteriana/fúngica en pacientes con COVID-19 ingresados en un hospital de tercer nivel de Castilla y León, España.

INTRODUCTION: Bacterial/fungal coinfection and superinfections contribute to the increased morbi-mortality of viral respiratory infections (RIs). The main objective of this study was to determine the incidence of these infections in hospitalized patients with COVID-19. METHOD: Retrospective observational study of all patients admitted for COVID-19 and bacterial/fungal infections at the Hospital Clínico Universitario of Valladolid, Spain (March 1-May 31, 2020). Demographic, clinical and microbiological data were compared based on Intensive Care Unit (ICU) admission and predictors of mortality by were identified using multivariate logistic regression analyses. RESULTS: Of the 712 COVID-19 patients, 113 (16%) presented bacterial/fungal coinfections or superinfections. Their median age was 73 years (IQR 57-89) and 59% were men. The profiles of ICU patients (44%) included male, SARS-CoV-2 pneumonia, leukocytosis, elevated inteleukin-6, with interferon ß-1b and tocilizumab and superinfection (p < 0.05). Coinfections were diagnosed in 5% (39/712) patients. Most common pathogens of respiratory coinfection (18) were Streptococcus pneumoniae (6) and Staphylococcus aureus (6). Superinfections were detected in 11% (80/712) patients. Urinary (53) and RIs (39) constituted the majority of superinfections Acinetobacter baumannii multidrug-resistant was the main agent of IR and bacteremia. An outbreak of A. baumannii contributed to this result. Three patients were considered to have probable pulmonary aspergillosis. Mortality was higher in UCI patients (50 vs. 29%; p = 0.028). The predictive factors of mortality included being a male with various comorbidities, SARS-CoV-2 pneumonia, bacteremia and superinfections from A. baumannii. CONCLUSION: The outbreak of A. baumannii was a determining factor in the increases of the incidence of infection and the morbi-mortality of ICU patients.

Tuberculosis por Mycobacterium bovis en la población de Castilla y León (España), 2006-2015 / Human Mycobacterium bovis infection in Castile and León (Spain), 2006-2015

Introducción: La incidencia anual de tuberculosis (TB) humana por Mycobacterium bovis ha disminuido considerablemente en los países industrializados desde inicios del sigloXX. El objetivo de este estudio fue conocer las características epidemiológicas, clínicas y microbiológicas de esta enfermedad en Castilla y León (CyL). Métodos: Estudio retrospectivo de los casos de TB por M. bovis de CyL en un periodo de 10 años, comparando la epidemiología, los factores de riesgo y la evolución entre las formas pulmonares (TBP) y extrapulmonares (TBEP). Resultados: Se recopilaron 75 casos de TB por M. bovis: 45 TBP y 31 TBEP. La incidencia acumulada de TB por M. bovis fue de 0,3 casos por 100.000 habitantes. Se mantuvo estable entre el primer y el segundo quinquenio (0,27 vs. 0,33, p = 0,656), a pesar del descenso de la incidencia global de la TB (13,58 vs. 10,71, p < 0,0001). La edad media fue de 66,2 + 21,3 años, principalmente varones (63%) y nacidos en España (92%). TBP fue significativamente más frecuente en varones, mayores de 66 años, con inmunosupresión o fumadores. La mortalidad fue del 9%, asociada a la edad, a la inmunosupresión o a un tratamiento diferente al recomendado por la OMS. Conclusiones: La incidencia de TB por M. bovis en CyL es superior a la de España y otros países europeos, y se mantuvo estable a pesar del descenso de la TB por MTC. Afectó mayoritariamente a población nacida en España que vivía en zonas rurales y con elevada media de edad

Introduction: The annual incidence of tuberculosis (TB) from Mycobacterium bovis in humans has considerably declined in industrialised countries since the early twentieth century. The objective of this study was to determine the epidemiological, clinical and microbiological characteristics of patients with this illness in Castile and León (CyL). Methods: Retrospective study of all M. bovis TB cases in CyL over a 10-year period, comparing the risk factors, the epidemiology and the clinical course between pulmonary (PTB) and extrapulmonary TB (EPTB). Results: 75 cases of TB were due to M. bovis: 45 PTB and 31 EPTB. The annual incidence of TB due to M. bovis was 0.3 cases per 100,000. It remained stable between the first and second five-year period (0.27 vs. 0.33, p = 0.656). However, the overall incidence of TB fell in both five-year periods (13.58 vs. 10.71, p < 0.0001). The mean age was 66.2 + 21.3 years, mainly men (63%) and Spanish patients (92%). PTB was significantly more frequent in men, aged over 66 years, with immunosuppressive conditions or who were smokers. Mortality was 9%, associated with higher age, immunosuppression or treatment different from that recommended by the WHO. Conclusions: The incidence of M. bovis TB in CyL was higher than that for Spain and for other European countries, and remained stable despite the decreased the TB due to MTC. It affected mostly Spanish-born patients who lived in rural areas and with a high mean age

Human Mycobacterium bovis infection in Castile and León (Spain), 2006-2015. / Tuberculosis por Mycobacterium bovis en la población de Castilla y León (España), 2006-2015.

INTRODUCTION: The annual incidence of tuberculosis (TB) from Mycobacterium bovis in humans has considerably declined in industrialised countries since the early twentieth century. The objective of this study was to determine the epidemiological, clinical and microbiological characteristics of patients with this illness in Castile and León (CyL). METHODS: Retrospective study of all M. bovis TB cases in CyL over a 10-year period, comparing the risk factors, the epidemiology and the clinical course between pulmonary (PTB) and extrapulmonary TB (EPTB). RESULTS: 75 cases of TB were due to M. bovis: 45 PTB and 31 EPTB. The annual incidence of TB due to M. bovis was 0.3 cases per 100,000. It remained stable between the first and second five-year period (0.27 vs. 0.33, p=0.656). However, the overall incidence of TB fell in both five-year periods (13.58 vs. 10.71, p<0.0001). The mean age was 66.2+21.3 years, mainly men (63%) and Spanish patients (92%). PTB was significantly more frequent in men, aged over 66 years, with immunosuppressive conditions or who were smokers. Mortality was 9%, associated with higher age, immunosuppression or treatment different from that recommended by the WHO. CONCLUSIONS: The incidence of M. bovis TB in CyL was higher than that for Spain and for other European countries, and remained stable despite the decreased the TB due to MTC. It affected mostly Spanish-born patients who lived in rural areas and with a high mean age.

Buruli ulcers in a Spanish aid worker after a stay in Peru.

Buruli ulcer (BU) is a chronic and destructive infection of the skin and soft tissues caused by Mycobacterium ulcerans. Recently, population flows have triggered the appearance of several sporadic cases of BU in non-endemic countries. This represents a significant diagnostic challenge for clinicians and microbiologists. We describe the first case of BU imported to Spain. The patient was a Spanish woman who had stayed 5 months in the jungle of Peru.

Síndrome de Lady Windermere en Castilla y León / Lady Windermere syndrome in Castile and León

INTRODUCCIÓN: El síndrome de Lady Windermere (SLW) es una patología pulmonar causada por Mycobacterium avium complex (MAC). El objetivo es conocer su frecuencia y sus características en el área norte de la comunidad de Castilla y León. MÉTODOS: Estudio retrospectivo de pacientes con aislamientos de MAC en muestras respiratorias de cinco hospitales públicos de la comunidad a lo largo de seis años, siguiendo criterios de la ATS/IDSA. Las cepas de MAC se identificaron por sondas de hibridación inversa Genotype Mycobacterium o PCR-RFLP del gen hsp65. RESULTADOS: De 183 casos de MAC identificados, únicamente 5 (2,7%) mujeres de 68,8 ± 10,7 años cumplían criterios de SLW. En tres casos se aisló MAC conjunta e intermitentemente con otros patógenos. Solo un paciente se trató siguiendo criterios de la ATS/IDSA. DISCUSIÓN: El SLW permanece infraestimado, y al ser los afectados muy demandantes de recursos sanitarios durante largos periodos, es necesario un mayor conocimiento microbiológico y terapéutico

INTRODUCTION: Lady Windermere syndrome (LWS) is a pulmonary disease caused by Mycobacterium avium complex (MAC). The objective of this study is to ascertain its frequency and characteristics in the northern area of the autonomous community of Castile and León. METHODS: A retrospective study of patients with MAC isolates in respiratory samples from five public hospitals in the autonomous community over a six-year period, following the ATS/IDSA criteria. The MAC strains were identified by GenoType Mycobacterium reverse hybridisation probes or PCR-RFLP analysis of the hsp65 gene. RESULTS: Of 183 cases of MAC identified, only five women (2.7 %) aged 68.8± 10.7 years met LWS criteria. In three cases, MAC was isolated jointly and intermittently with other pathogens. Only one patient was treated according to ATS/IDSA criteria. DISCUSSION: LWS remains underestimated, with affected patients representing a significant burden on healthcare resources over long periods of time. As a result, greater microbiological and therapeutic knowledge of the syndrome is needed

Contribución de la microbiología al diagnóstico de la tuberculosis en Castilla y León: conclusiones del estudio GRUMICALE 2013 / Contribution of microbiology in the diagnosis of tuberculosis in Castile and León (Spain): Findings of the GRUMICALE 2013 study

Introducción y objetivos: Estudio retrospectivo que recoge datos microbiológicos de tuberculosis (TB) en Castilla y León durante el año 2013 para conocer los datos microbiológicos de incidencia y distribución de TB, resistencias a los fármacos antituberculosos y compararlos con los datos epidemiológicos ofrecidos por los servicios de vigilancia epidemiológica (SIVE). Material y métodos: Microbiólogos de los 14 hospitales de la red sanitaria pública de Castilla y León (GRUMICALE) han recogido datos epidemiológicos, microbiológicos y de funcionamiento de los laboratorios de microbiología de la comunidad durante el año 2013. Se consideró un solo aislamiento de Mycobacterium tuberculosis complex (MTC) por paciente. Resultados: Se obtuvieron 270 aislamientos de MTC (tasa de incidencia de 11,63 casos/100.000 hab./año). Según datos epidemiológicos, se recogieron un total de 288 casos de TB (11,43 casos/100.000 hab./año), 243 confirmados, 29 sospechosos y 16 probables. Predomina la localización pulmonar, seguida de lejos por la pleural y por el resto. Se procesaron un total de 27.620 muestras para micobacterias. En un 3,46% de los medios de cultivos líquidos se obtuvo crecimiento de micobacterias, y en un 50,37% la tinción directa (baciloscopia) fue positiva. Dieciséis aislamientos de Mycobacterium tuberculosis (MT) presentaron resistencia a algún fármaco antituberculoso, predominando la resistencia a isoniazida (5,92%). La provincia con mayor incidencia y número de aislamientos fue León (24,23 casos/100.000 hab./año), siendo la máxima en el área sanitaria de El Bierzo (30,46 casos/100.000 hab./año). Conclusiones: Una adecuada recogida de la información microbiológica es fundamental para el conocimiento de la epidemiologia de la TB en nuestra comunidad (AU)

Introduction and objectives: A retrospective study was conducted by collecting microbiological tuberculosis (TB) data in Castile and León during the year 2013 in order to determine the incidence and distribution of TB, and resistance to the tuberculostatic drug, and compare them with the epidemiological data provided by the Department of Epidemiological Surveillance (SIVE). Material and methods: Microbiologists of the 14 hospitals of the Castile and León public health network (GRUMICALE) collected epidemiological, microbiological, and management data from the Microbiology laboratories in the community during the year 2013. A single isolate of Mycobacterium tuberculosis complex (MTC) per patient was considered. Results: The study included a total of 270 MTC isolates (an incidence rate of 11.63 cases/100,000 inhab./year). A total of 288 cases of TB (11.43 cases/100,000 inhab. year) were recovered using epidemiological data, which included 243 confirmed, 29 suspected, and 16 as probable cases. Pulmonary TB was predominant, followed a long way off by the pleural TB and the remaining locations. A total of 27,620 samples were processed for mycobacterial detection. Mycobacterial growth was observed in 3.46% of automated fluid cultures, and 50.37% were positive by direct staining of the smear. Resistance to one tuberculostatic drug, mostly to isoniazid, was observed in 16 (5.92%) isolates of Mycobacterium tuberculosis (MT). The province with greater incidence and number of isolates was León (24.23 cases/100,000 inhab./year), with the highest being observed in El Bierzo health area (30.46 cases/100,000 inhab./year). Conclusions: An adequate collection of microbiological information is essential to determine the epidemiology of TB in our region (AU)

Lady Windermere syndrome in Castile and León. / Síndrome de Lady Windermere en Castilla y León.

INTRODUCTION: Lady Windermere syndrome (LWS) is a pulmonary disease caused by Mycobacterium avium complex (MAC). The objective of this study is to ascertain its frequency and characteristics in the northern area of the autonomous community of Castile and León. METHODS: A retrospective study of patients with MAC isolates in respiratory samples from five public hospitals in the autonomous community over a six-year period, following the ATS/IDSA criteria. The MAC strains were identified by GenoType Mycobacterium reverse hybridisation probes or PCR-RFLP analysis of the hsp65 gene. RESULTS: Of 183 cases of MAC identified, only five women (2.7%) aged 68.8±10.7years met LWS criteria. In three cases, MAC was isolated jointly and intermittently with other pathogens. Only one patient was treated according to ATS/IDSA criteria. DISCUSSION: LWS remains underestimated, with affected patients representing a significant burden on healthcare resources over long periods of time. As a result, greater microbiological and therapeutic knowledge of the syndrome is needed.

Contribution of microbiology in the diagnosis of tuberculosis in Castile and León (Spain): Findings of the GRUMICALE 2013 study. / Contribución de la microbiología al diagnóstico de la tuberculosis en Castilla y León: conclusiones del estudio GRUMICALE 2013.

INTRODUCTION AND OBJECTIVES: A retrospective study was conducted by collecting microbiological tuberculosis (TB) data in Castile and León during the year 2013 in order to determine the incidence and distribution of TB, and resistance to the tuberculostatic drug, and compare them with the epidemiological data provided by the Department of Epidemiological Surveillance (SIVE). MATERIAL AND METHODS: Microbiologists of the 14 hospitals of the Castile and León public health network (GRUMICALE) collected epidemiological, microbiological, and management data from the Microbiology laboratories in the community during the year 2013. A single isolate of Mycobacterium tuberculosis complex (MTC) per patient was considered. RESULTS: The study included a total of 270 MTC isolates (an incidence rate of 11.63 cases/100,000 inhab./year). A total of 288 cases of TB (11.43 cases/100,000 inhab. year) were recovered using epidemiological data, which included 243 confirmed, 29 suspected, and 16 as probable cases. Pulmonary TB was predominant, followed a long way off by the pleural TB and the remaining locations. A total of 27,620 samples were processed for mycobacterial detection. Mycobacterial growth was observed in 3.46% of automated fluid cultures, and 50.37% were positive by direct staining of the smear. Resistance to one tuberculostatic drug, mostly to isoniazid, was observed in 16 (5.92%) isolates of Mycobacterium tuberculosis (MT). The province with greater incidence and number of isolates was León (24.23 cases/100,000 inhab./year), with the highest being observed in El Bierzo health area (30.46 cases/100,000 inhab./year). CONCLUSIONS: An adequate collection of microbiological information is essential to determine the epidemiology of TB in our region.

In vitro infection with Mycobacterium tuberculosis induces a distinct immunological pattern in blood from healthy relatives of tuberculosis patients.

Part of the susceptibility to tuberculosis has a genetic basis, which is clear in primary immunodeficiencies, but is less evident in apparently immunocompetent subjects. Immune responses were analysed in blood samples from tuberculosis patients and their healthy first-degree relatives who were infected in vitro with mycobacteria (either Mycobacterium tuberculosis or M. bovis BCG). The antimicrobial activity against M. tuberculosis in blood from relatives was significantly lower than that observed in healthy controls. Tuberculosis patients exhibited a higher number of neutrophils, and monocyte phagocytosis was inhibited in both relatives and tuberculosis patients. A remarkable finding was that the production of reactive oxygen species by infected neutrophils was higher in relatives than in healthy controls. A higher production of TNFα in infected blood from relatives was also observed. These results may indicate that relatives display a stronger inflammatory response and that their immune response to M. tuberculosis is different from those of unrelated controls. First-degree relatives may represent a highly informative group for the analysis of tuberculosis susceptibility.

Blood antimicrobial activity varies against different Mycobacterium spp.

In vitro analysis of mycobacterial pathogenicity or host susceptibility has traditionally relied on the infection of macrophages, the target cell of mycobacteria, despite difficulties reproducing their antimycobacterial activity. We have employed alternative models, namely whole blood and leukocytes in plasma, from QuantiFERON negative individuals, and performed infections with the pathogenic M. tuberculosis, the less pathogenic M. avium, M. kansasii and M. chelonae and the occasionally pathogenic M. gordonae and M. bovis. The anticoagulant used in blood extraction, heparin or EDTA, had a major influence in the outcome of the infection. Thus, while in the heparinized models a similar number of bacteria were enumerated in the inoculum and after seven days, in the presence of EDTA a killing effect was observed, despite the inhibitory effect of EDTA on cellular functions like the production of cytokines or reactive oxygen species (ROS). A special case was the rapidly growing mycobacteria M. chelonae, that multiplied in heparinized models but was eliminated in models with EDTA. We verified that EDTA is not responsible for the bactericidal effect, but acts as a bacteriostatic agent. Further work will determine whether blood derived models are a better alternative to the classical macrophage.

Immunological response to Mycobacterium tuberculosis infection in blood from type 2 diabetes patients.

The convergence of tuberculosis and diabetes represents a co-epidemic that threatens progress against tuberculosis. We have investigated type 2 diabetes as a risk factor for tuberculosis susceptibility, and have used as experimental model whole blood infected in vitro with Mycobacterium tuberculosis. Blood samples from diabetic patients were found to have a higher absolute neutrophil count that non-diabetic controls, but their immune functionality seemed impaired because they displayed a lower capacity to phagocytose M. tuberculosis, a finding that had been previously reported only for monocytes. In contrast, an increased production of TNFα was detected in infected blood from diabetic patients. Despite the altered phagocytic capacity showed by cells from these patients, the antimicrobial activity measured in both whole blood and monocyte derived macrophages was similar to that of controls. This unexpected result prompts further improvements in the whole blood model to analyze the immune response of diabetes patients to tuberculosis.