Distribution of Aspergillus species and prevalence of azole resistance in clinical and environmental samples from a Spanish hospital during a three-year study period.

BACKGROUND: Surveillance studies are crucial for updating trends in Aspergillus species and antifungal susceptibility information. OBJECTIVES: Determine the Aspergillus species distribution and azole resistance prevalence during this 3-year prospective surveillance study in a Spanish hospital. MATERIALS AND METHODS: Three hundred thirty-five Aspergillus spp. clinical and environmental isolates were collected during a 3-year study. All isolates were screened for azole resistance using an agar-based screening method and resistance was confirmed by EUCAST antifungal susceptibility testing. The azole resistance mechanism was confirmed by sequencing the cyp51A gene and its promoter. All Aspergillus fumigatus strains were genotyped using TRESPERG analysis. RESULTS: Aspergillus fumigatus was the predominant species recovered with a total of 174 strains (51.94%). The rest of Aspergillus spp. were less frequent: Aspergillus niger (14.93%), Aspergillus terreus (9.55%), Aspergillus flavus (8.36%), Aspergillus nidulans (5.37%) and Aspergillus lentulus (3.28%), among other Aspergillus species (6.57%). TRESPERG analysis showed 99 different genotypes, with 72.73% of the strains being represented as a single genotype. Some genotypes were common among clinical and environmental A. fumigatus azole-susceptible strains, even when isolated months apart. We describe the occurrence of two azole-resistant A. fumigatus strains, one clinical and another environmental, that were genotypically different and did not share genotypes with any of the azole-susceptible strains. CONCLUSIONS: Aspergillus fumigatus strains showed a very diverse population although several genotypes were shared among clinical and environmental strains. The isolation of azole-resistant strains from both settings suggest that an efficient analysis of clinical and environmental sources must be done to detect azole resistance in A. fumigatus.

Summation of peaks and L34 ribosomal protein in the presence and absence of antibiotics enables susceptibility testing using MALDI-TOF mass spectrometry in 2h from Escherichia coli-positive blood cultures / El sumatorio de picos y la proteína ribosómica L34, en presencia y ausencia de antimicrobianos, permiten conocer la sensibilidad a antimicrobianos mediante espectrometría de masas MALDI-TOF en 2h, en hemocultivos positivos para Escherichia coli

INTRODUCTION: We have developed a MALDI-TOF-mediated phenotypic method, which determines antibiotic susceptibility (AS) from positive blood cultures (BCs) in 2h. We developed a software for process automation. We report results on Escherichia coli-positive BCs with cefotaxime (CTX) and ciprofloxacin (CIP). METHODS: We studied CIP and CTX activity in 18 and 17 real E. coli-positive BCs, and in 56 and 45 spiked BCs, respectively. Positive BCs were incubated for 2 h without any antibiotics, and with 2 mg/l and 4 mg/l of CIP and CTX. The extraction was performed using ethanol/formic acid. Spectra were processed with specifically developed software which compares the peaks' intensity and the size of specific peaks. RESULTS: The set cut-off point was a 3-fold decrease in the summation of all peaks and/or the 5382 m/z peak value (ribosomal protein L34). In simulated BCs, the correlation of CIP 2mg/l and 4mg/l with Etest(R) was 94.6% and 98.2%, respectively; for CTX 2 mg/l and 4 mg/l, this correlation was 95.6%. In real BCs, the correlations were 100% for CIP (2mg/l and 4mg/l) and 88.2% and 94.1% for CTX 2 mg/l and 4 mg/l, respectively. Resistant isolates were always correctly classified. CONCLUSION: This method provides accurate, fast and inexpensive AS information. The method can be automated, making it easier to implement in a microbiology laboratory routine

INTRODUCCIÓN: Se ha desarrollado un método fenotípico basado en MALDI-TOF, que determina la sensibilidad a antibióticos en hemocultivos (HC) positivos en 2h. Se ha desarrollado un software que automatiza el proceso. Se presentan los resultados en HC positivos para Escherichia coli, con cefotaxima (CTX) y ciprofloxacino (CIP). MÉTODOS: Se estudió la actividad de CIP y CTX en 18 y 17HC positivos reales con E. coli, y en 56 y 45 HC simulados. Los HC positivos se incubaron durante 2h sin antibiótico, y con 2 y 4 mg/l de CIP y de CTX. La extracción se realizó con etanol/ácido fórmico. Los espectros se procesaron con un software específico, que compara la intensidad de los picos y el tamaño de los picos específicos. RESULTADOS: El punto de corte establecido fue una disminución de 3 veces en la suma de picos, y/o en el valor del pico de 5.382 m/z (proteína ribosómica L34). En hemocultivos simulados la correlación con Etest(R) para las concentraciones de CIP de 2 y 4 mg/l fueron 94,6 y 98,2%, respectivamente, y 95,6% para CTX (2 y 4 mg/l). En HC reales, la correlación con Etest(R) fue del 100% para CIP (2 y 4 mg/l), y del 88,2 y 94,1% para CTX 2 y 4 mg/l, respectivamente. Los aislados resistentes siempre se clasificaron correctamente. CONCLUSIÓN: Este método proporciona información sobre sensibilidad a antimicrobianos de manera precisa, rápida y barata. El método se puede automatizar e incluir en la rutina del laboratorio de microbiología

Summation of peaks and L34 ribosomal protein in the presence and absence of antibiotics enables susceptibility testing using MALDI-TOF mass spectrometry in 2h from Escherichia coli-positive blood cultures.

INTRODUCTION: We have developed a MALDI-TOF-mediated phenotypic method, which determines antibiotic susceptibility (AS) from positive blood cultures (BCs) in 2h. We developed a software for process automation. We report results on Escherichia coli-positive BCs with cefotaxime (CTX) and ciprofloxacin (CIP). METHODS: We studied CIP and CTX activity in 18 and 17 real E. coli-positive BCs, and in 56 and 45 spiked BCs, respectively. Positive BCs were incubated for 2h without any antibiotics, and with 2mg/l and 4mg/l of CIP and CTX. The extraction was performed using ethanol/formic acid. Spectra were processed with specifically developed software which compares the peaks' intensity and the size of specific peaks. RESULTS: The set cut-off point was a 3-fold decrease in the summation of all peaks and/or the 5382m/z peak value (ribosomal protein L34). In simulated BCs, the correlation of CIP 2mg/l and 4mg/l with Etest® was 94.6% and 98.2%, respectively; for CTX 2mg/l and 4mg/l, this correlation was 95.6%. In real BCs, the correlations were 100% for CIP (2mg/l and 4mg/l) and 88.2% and 94.1% for CTX 2mg/l and 4mg/l, respectively. Resistant isolates were always correctly classified. CONCLUSION: This method provides accurate, fast and inexpensive AS information. The method can be automated, making it easier to implement in a microbiology laboratory routine.

Parásitos e inmigración: ¿cuál es la mejor actitud? / Parasites and immigration: what is the best approach?

El aumento de la población inmigrante en Europa (en España se considera que supone, en este momento, el 10% de la población total) conlleva un incremento en la prevalencia de determinadas enfermedades, en especial enfermedades infecciosas, y obliga a enfocar el estudio de los pacientes teniendo muy en cuenta las enfermedades prevalentes en su zona de origen. Diferentes organismos defienden el cribado sistemático, o el tratamiento empírico indiscriminado. El cribado debe incluir, al menos, un estudio coproparasitológico, de hemoparásitos y de parásitos urinarios. El estudio coproparasitológico es esencial, dada la alta prevalencia de parásitos intestinales en población procedente de áreas tropicales y subtropicales. La búsqueda de hemoparásitos es fundamental para la detección de Plasmodium, Babesia, Trypanosoma y también de algunas filarias. En individuos asintomáticos procedentes de zonas endémicas para Plasmodium es primordial la introducción de técnicas moleculares, dada la baja sensibilidad del resto. Todo individuo procedente de áreas endémicas para filariasis o enfermedad de Chagas debe ser sometido al cribado de estas. El cribado de Trypanosoma cruzi es de especial relevancia en embarazadas y donantes de sangre y de órganos. Se revisan los agentes etiológicos a tener en cuenta en población inmigrante ante un síndrome febril, cardiovascular, respiratorio, digestivo, hepatoesplenomegalia, síndromes nefrourológicos, neurológicos, reumatológicos, oftalmológicos, otorrinolaringológicos y cutáneos (AU)

The increasing immigrant population in Europe (in Spain it is assumed to account for 10% of the total population) is leading to a higher prevalence of some diseases, especially infectious diseases. When studying these patients, the most prevalent diseases in their area of origin must be taken into account. Different agencies advocate systematic screening, or indiscriminate empirical treatment. Screening should include a search for parasites in stool, blood and urine samples. Stool parasitological studies are fundamental, given the high prevalence of intestinal parasites in populations from tropical and subtropical areas. The search for blood parasites is essential for the detection of Plasmodium, Babesia, Trypanosoma and also some filaria. In asymptomatic individuals coming from areas endemic for Plasmodium, the use of molecular techniques is necessary, given the low sensitivity of conventional methods. All individuals from areas endemic for filariasis or Chagas disease should be screened. Screening for Trypanosoma cruzi is particularly important in pregnant women and blood and organ donors. We review the main agents to be considered in immigrant patients showing febrile, cardiovascular, respiratory or gastrointestinal syndromes, hepatosplenomegaly, and nephro-urological, neurological, rheumatologic, ophthalmological, ear, nose and throat (ENT) and cutaneous síndromes (AU)

Métodos rápidos de identificación de bacterias y hongos. Espectrometría de masas MALDI-TOF, medios cromogénicos / Fast methods of fungal and bacterial identification. MALDI-TOF mass spectrometry, chromogenic media

La espectrometría de masas MALDI-TOF es ya una herramienta de trabajo rutinaria en Microbiología Clínica, por su rapidez y fiabilidad en la identificación de microorganismos. Sus resultados están perfectamente contrastados en la identificación de bacterias y levaduras. La identificación de micobacterias y hongos filamentosos presenta mayor complejidad, por la mayor heterogeneidad de espectros dentro de cada especie. La metodología es algo más compleja, y la ampliación del número de especies de referencia, y del número de espectros de cada especie, serán cruciales para alcanzar mayor eficacia. La identificación directa a partir de hemocultivos se ha implantado dada su aportación al manejo de pacientes graves, pero su aplicación a otras muestras es más compleja. Los medios de cultivos cromogénicos han supuesto también una aportación al diagnóstico rápido tanto en bacterias como en levaduras, ya que aceleran el diagnóstico, facilitan la detección de cultivos mixtos y permiten un diagnóstico rápido de especies resistentes

MALDI-TOF mass spectrometry is now a routine resource in Clinical Microbiology, because of its speed and reliability in the identification of microorganisms. Its performance in the identification of bacteria and yeasts is perfectly contrasted. The identification of mycobacteria and moulds is more complex, due to the heterogeneity of spectra within each species. The methodology is somewhat more complex, and expanding the size of species libraries, and the number of spectra of each species, will be crucial to achieve greater efficiency. Direct identification from blood cultures has been implemented, since its contribution to the management of severe patients is evident, but its application to other samples is more complex. Chromogenic media have also contributed to the rapid diagnosis in both bacteria and yeast, since they accelerate the diagnosis, facilitate the detection of mixed cultures and allow rapid diagnosis of resistant species

Unusual clinical presentations of Actinotignum (Actinobaculum) schaalii infection / Presentaciones clínicas inusuales de la infección por Actinotignum (Actinobaculum) schaalii

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Fast methods of fungal and bacterial identification. MALDI-TOF mass spectrometry, chromogenic media. / Métodos rápidos de identificación de bacterias y hongos. Espectrometría de masas MALDI-TOF, medios cromogénicos.

MALDI-TOF mass spectrometry is now a routine resource in Clinical Microbiology, because of its speed and reliability in the identification of microorganisms. Its performance in the identification of bacteria and yeasts is perfectly contrasted. The identification of mycobacteria and moulds is more complex, due to the heterogeneity of spectra within each species. The methodology is somewhat more complex, and expanding the size of species libraries, and the number of spectra of each species, will be crucial to achieve greater efficiency. Direct identification from blood cultures has been implemented, since its contribution to the management of severe patients is evident, but its application to other samples is more complex. Chromogenic media have also contributed to the rapid diagnosis in both bacteria and yeast, since they accelerate the diagnosis, facilitate the detection of mixed cultures and allow rapid diagnosis of resistant species.

Screening for parasite infections in immigrant children from low-income countries / Detección de infecciones parasitarias en menores inmigrantes procedentes de países en vías de desarrollo

INTRODUCTION: In Spain, minors represent approximately 20% of the immigration flow. Many of these immigrants come from countries in the tropics and sub-tropics where intestinal parasitic infections caused by helminths and protozoa are one of the major causes of human disease. The main objective of the present work was to describe parasite infections in a group of immigrant children. METHODS: A prospective evaluation was performed in 373 minors from Sub-Saharan Africa, North Africa, and Latin America. Details were collected from the medical records and physical examination. Urine, stool and peripheral blood samples were obtained for serological and routine laboratory tests. Direct and indirect parasitological tests were also performed. RESULTS: At least 1 parasitic disease was diagnosed in 176 (47.1%) immigrant children, while 77 (20.6%) minors were infected with two or more parasites. The number of parasites was highest in children from Sub-Saharan Africa compared with the rest of the areas of origin (p<.001), and in children from urban areas compared with those from rural areas (OR 1.27 [1.059-1.552], p=.011). The most frequent causes of multiple parasite infection were filariasis plus strongyloidiasis and filariasis plus schistosomiasis. Intestinal parasite infection was diagnosed in 38 cases (13.8%). Logistic regression analysis revealed that for each month of stay, the probability of a positive finding in the stool sample decreased by 0.02% [β=−0.020, (p=.07)]. CONCLUSIONS: The high infection rates of parasite diseases in immigrant children point to the need for screening protocols for certain infectious diseases in these children according to their country of origin and their length of residence in Spain

INTRODUCCIÓN: En España, los menores representan aproximadamente el 20% del flujo migratorio. Muchos de estos menores provienen de regiones tropicales y subtropicales donde las infecciones por helmintos y protozoos son una de las principales causas de morbilidad. El objetivo de este trabajo es describir las infecciones parasitarias presentes en un colectivo de menores inmigrantes. MÉTODOS: Se evaluaron prospectivamente 373 menores procedentes de África subsahariana, África del Norte y Latinoamérica. Se realizó una historia clínica detallada. Se obtuvieron muestras de sangre periférica, orina y heces para la realización de los diferentes análisis bioquímicos, serológicos y parasitológicos directos e indirectos. RESULTADOS: En 176 (47,1%) menores se diagnosticó al menos una enfermedad parasitaria. En 77 (20,6%) menores se detectaron 2 o más parásitos. En los niños de África subsahariana el número de parásitos fue mayor comparado el resto de orígenes (p < 0,001). Los menores de zonas urbanas tenían más parásitos comparado con los niños de zonas rurales (OR 1,27 [1059-1552], p = 0,011). Las causas más frecuentes de parasitación múltiple fueron filariosis más estrongiloidosis y filariosis más esquistosomiasis. Se diagnosticó parasitosis intestinal en 38 casos (13,8%). El análisis de regresión logística reveló que por cada mes de estancia, la probabilidad de un resultado positivo en las heces disminuía un 0,02% [β=−0,020 (p = 0,07)]. CONCLUSIÓN: Las altas tasas de infección parasitaria en niños inmigrantes señala la necesidad de una detección protocolizada de estas enfermedades según el país de origen y el tiempo de residencia en España

Screening for parasite infections in immigrant children from low-income countries.

INTRODUCTION: In Spain, minors represent approximately 20% of the immigration flow. Many of these immigrants come from countries in the tropics and sub-tropics where intestinal parasitic infections caused by helminths and protozoa are one of the major causes of human disease. The main objective of the present work was to describe parasite infections in a group of immigrant children. METHODS: A prospective evaluation was performed in 373 minors from Sub-Saharan Africa, North Africa, and Latin America. Details were collected from the medical records and physical examination. Urine, stool and peripheral blood samples were obtained for serological and routine laboratory tests. Direct and indirect parasitological tests were also performed. RESULTS: At least 1 parasitic disease was diagnosed in 176 (47.1%) immigrant children, while 77 (20.6%) minors were infected with two or more parasites. The number of parasites was highest in children from Sub-Saharan Africa compared with the rest of the areas of origin (p<.001), and in children from urban areas compared with those from rural areas (OR 1.27 [1.059-1.552], p=.011). The most frequent causes of multiple parasite infection were filariasis plus strongyloidiasis and filariasis plus schistosomiasis. Intestinal parasite infection was diagnosed in 38 cases (13.8%). Logistic regression analysis revealed that for each month of stay, the probability of a positive finding in the stool sample decreased by 0.02% [ß=-0.020, (p=.07)]. CONCLUSIONS: The high infection rates of parasite diseases in immigrant children point to the need for screening protocols for certain infectious diseases in these children according to their country of origin and their length of residence in Spain.

Características clínico-demográficas de la población del Sur de Gran Canaria con diagnóstico de enterobiasis: valoración de la toma de muestras / Demographic and clinical features of diagnosed individuals of enterobiasis in the southern Gran Canaria: sampling assessment

Introducción. La enterobiasis, causada por Enterobius vermicularis es una de las parasitosis más prevalentes en población infantil, cuyo diagnóstico implica la demostración de los huevos o gusanos mediante el método de Graham. El objetivo del estudio es describir las características clínico-demográficas y microbiológicas de pacientes con sospecha de Enterobiasis en el área Sur de Gran Canaria. Material y métodos. Estudio descriptivo y prospectivo de los resultados de muestras perianales evaluadas mediante el método de Graham por el Servicio de Microbiología del Hospital Universitario Insular de Gran Canaria entre Noviembre de 2014 y Noviembre de 2015. Se ha realizado un análisis descriptivo y de asociación de riesgo de las variables clínicas y demográficas y los resultados del Test de Graham. Resultados. Se obtuvieron 1.128 muestras válidas. En el 11,4% se observaron huevos de E. vermicularis; el 88,4% de las muestras positivas en menores de 14 años y el 53,5% en género masculino. Dolor abdominal (18,6%), prurito anal (11,6%), eosinofilia (8,5%) y parasitosis intestinal (7,8%) fueron los motivos de solicitud en las muestras positivas. Predomino elevado sin diagnóstico de sospecha o diagnósticos no relacionados con enterobiasis. Conclusiones. La enterobiasis es un motivo de consulta en atención primaria y una patología de interés en Gran Canaria. La calidad de recogida de muestras y el diagnóstico de sospecha es necesaria para realizar un buen análisis microbiológico (AU)

Introduction. Enterobius vermicularis, also known as pinworn, is the responsible agent for Human Enterobiasis. It is one of the most prevalent, but underrated, parasitic disease in children population. Diagnosis involves demonstration of either eggs or adult worms by Graham test. The aim of this study is to describe the clinical, demographic and microbiological features of patients with suspected diagnosis of Enterobiasis in southern Gran Canaria. Material and methods. Descriptive and prospective study of perianal samples evaluated by Graham test in the Microbiology Department of `Insular de Gran Canaria´ University Hospital between November 2014 and November 2015. Descriptive analysis to evaluate the correlation between clinical and demographic variables and the results of Graham test microbiological observation. Results. 1,128 samples were analyzed. E. vermicularis was found in 11.4% of the samples. Among the positives samples, 88.4% belonged to children under 14 years, and 53.5% were male. Abdominal pain (18.6%), anal itching (11.6%), eosinophilia (8.5%) and intestinal parasitosis suspicion (7.8%) were the reasons of parasitological investigation request in positive samples. Nevertheless, a high proportion of the requests was not founded in a suspicious diagnosis or was unrelated to Enterobiasis. Conclusions. Enterobiasis is a common disease in primary health care and is of great importance in Gran Canaria. Quality in sample collection as well as diagnosis suspicious information are necessary for a good microbiological analysis (AU)

Peritonitis por Moraxella osloensis: a propósito de un caso y revisión de la literatura / Moraxella osloensis peritonitis: case report and review

No disponible

[Epidemiology of Clostridium difficile-associated disease (CDAD) in Salamanca]. / Epidemiología de la Enfermedad Asociada a Clostridium difficile (EACD) en Salamanca.

INTRODUCTION: Clostridium difficile infection is considered a major cause of nosocomial diarrhoea in developed countries and is increasingly becoming more important as an etiologic agent of community diarrhoea, also in patients without risk factors. METHOD: Beginning in May 2011, the aim of our study is to know the characteristics of patients suffering from C. difficile Associated Disease in Salamanca University Hospital, collecting their data in a survey conducted for this purpose. A case was defined as a patient with compatible clinical and positive microbiological diagnosis. RESULTS: After 18 months of study, 41 cases had been documented representing an incidence of 1.15 cases per 10,000 patient-days. Patients were hospitalized (37) or health care associated (4), females (54%), age ≥ 65 years (56%) with prior antibiotic treatment (80%), most had diarrhea after the third day of admission, less than three weeks and without blood. Most were treated with metronidazole alone (78%), 19% with metronidazole and vancomycin, and the remaining percentage was resolved without treatment. Recurrences were about 20% and 7 (17%) died. CONCLUSIONS: The characteristics of our patients with C. difficile - associated disease are the same as those reported by other authors. Local surveillance is important in order to study the endemic and epidemic C. difficile infection. According to published epidemiological changes, we should be able to develop strategies from the Microbiology laboratories that will improve diagnosis of the disease.

Epidemiología de la Enfermedad Asociada a Clostridium difficile (EACD) en Salamanca / Epidemiology of Clostridium difficile-associated disease (CDAD) in Salamanca

Introducción. La infección por Clostridium difficile se considera la principal causa de diarrea nosocomial en países desarrollados y cada vez cobra más relevancia como agente etiológico de diarreas comunitarias, y en pacientes sin factores considerados de riesgo. Método. En este estudio que comienza en Mayo de 2011 planteamos conocer las características de los pacientes afectados de Enfermedad Asociada a C. difficile en el Complejo Asistencial Universitario de Salamanca, recogiendo sus datos en una encuesta elaborada a tal efecto. Se consideró como caso el paciente con clínica compatible y diagnóstico microbiológico positivo. Resultados. A los 18 meses del comienzo se habían documentado 41 casos lo que supone una incidencia de 1.15 casos por 10.000 pacientes-día. Fueron pacientes hospitalizados (37) o relacionados con asistencia sanitaria (4), mujeres (54%), añosas (56%) con tratamiento antibiótico previo (80%), la mayoría presentaron diarrea tras el tercer día de ingreso, de menos de tres semanas y sin sangre. La mayoría fueron tratados solo con metronidazol (78%), un 19% con metronidazol y vancomicina asociados y el restante se resolvió sin tratamiento. Recayeron cerca de un 20% y 7 (17%) fallecieron. Conclusiones. Las características de nuestros pacientes con Enfermedad Asociada a C. difficile son las mismas que reportan otros autores. Es importante la vigilancia local para conocer la endemia y vigilar las modificaciones no esperables de la incidencia. Teniendo en cuenta los cambios epidemiológicos que reporta la bibliografía, desde Microbiología debemos trabajar en estrategias que optimicen el diagnóstico de esta enfermedad (AU)

Introduction. Clostridium difficile infection is considered a major cause of nosocomial diarrhoea in developed countries and is increasingly becoming more important as an etiologic agent of community diarrhoea, also in patients without risk factors. Method. Beginning in May 2011, the aim of our study is to know the characteristics of patients suffering from C. difficile Associated Disease in Salamanca University Hospital, collecting their data in a survey conducted for this purpose. A case was defined as a patient with compatible clinical and positive microbiological diagnosis. Results. After 18 months of study, 41 cases had been documented representing an incidence of 1.15 cases per 10,000 patient-days. Patients were hospitalized (37) or health care associated (4), females (54%), age ≥65 years (56%) with prior antibiotic treatment (80%), most had diarrhea after the third day of admission, less than three weeks and without blood. Most were treated with metronidazole alone (78%), 19% with metronidazole and vancomycin, and the remaining percentage was resolved without treatment. Recurrences were about 20% and 7 (17%) died. Conclusions. The characteristics of our patients with C. difficile - associated disease are the same as those reported by other authors. Local surveillance is important in order to study the endemic and epidemic C. difficile infection. According to published epidemiological changes, we sould be able to develop strategies from the Microbiology laboratories that will improve diagnosis of the disease (AU)