RESUMEN
Candida albicans is the most frequent yeast responsible for systemic infections in humans. These infections mainly originate from the gastrointestinal tract where C. albicans can invade the gut epithelial barrier to gain access to the bloodstream. Along the gut, pathogens can use Microfold (M) cells as a portal of entry to cross the epithelial barrier. M cells are specialized cells mainly located in the follicule-associated epithelium of Peyer patches. In this study, we used scanning electron and fluorescence microscopy, adhesion and invasion assays and fungal mutants to investigate the interactions of C. albicans with M cells obtained in an established in vitro model whereby enterocyte-like Caco-2 cells co-cultured with the Raji B cell line undergo a phenotypic switch to morphologically and functionally resembling M cells. Our data demonstrate that C. albicans co-localizes with and invades preferentially M cells, providing evidence that the fungus can use M cells as a portal of entry into the intestinal barrier. In addition to active penetration, F-actin dependent endocytosis contributes to internalization of the fungus into M cells through a mechanism involving hypha-associated invasins including Ssa1 and Als3.
Asunto(s)
Candida albicans/fisiología , Candidemia/microbiología , Tracto Gastrointestinal/microbiología , Interacciones Huésped-Patógeno , Ganglios Linfáticos Agregados/microbiología , Linfocitos B/fisiología , Adhesión Celular , Línea Celular , Técnicas de Cocultivo , Endocitosis , Células Epiteliales/microbiología , Células Epiteliales/fisiología , Humanos , Microscopía Electrónica de Rastreo , Microscopía FluorescenteRESUMEN
Fusarium oxysporum is typically a soilborne fungus but can also be found in aquatic environments. In hospitals, water distribution systems may be reservoirs for the fungi responsible for nosocomial infections. F. oxysporum was previously detected in the water distribution systems of five French hospitals. Sixty-eight isolates from water representative of all hospital units that were previously sampled and characterized by translation elongation factor 1α sequence typing were subjected to microsatellite analysis and full-length ribosomal intergenic spacer (IGS) sequence typing. All but three isolates shared common microsatellite loci and a common two-locus sequence type (ST). This ST has an international geographical distribution in both the water networks of hospitals and among clinical isolates. The ST dominant in water was not detected among 300 isolates of F. oxysporum that originated from surrounding soils. Further characterization of 15 isolates by vegetative compatibility testing allowed us to conclude that a clonal lineage of F. oxysporum circulates in the tap water of the different hospitals. IMPORTANCE: We demonstrated that a clonal lineage of Fusarium oxysporum inhabits the water distribution systems of several French hospitals. This clonal lineage, which appears to be particularly adapted to water networks, represents a potential risk for human infection and raises questions about its worldwide distribution.
Asunto(s)
Agua Potable/microbiología , Fusarium/genética , Fusarium/aislamiento & purificación , Hospitales , ADN de Hongos/aislamiento & purificación , ADN Intergénico , Francia/epidemiología , Fusariosis/epidemiología , Fusariosis/etiología , Fusariosis/microbiología , Fusarium/clasificación , Humanos , Repeticiones de Microsatélite , Factor 1 de Elongación Peptídica/genética , Filogenia , Análisis de Secuencia de ADNRESUMEN
Conventional identification (CI) of yeasts is based on morphological, biochemical and/or immunological methods. Matrix-assisted laser desorption/ionization - time of flight (MALDI-TOF or MT-MS) mass spectrometry has been proposed as a new method for the identification of microorganisms. This prospective study compared the performance of MT-MS and CI for the identification of yeasts isolated from clinical samples. Sequencing of the internal transcribed spacer (ITS) regions of ribosomal DNA was used as the reference method in the analysis of a total of 1207 yeast isolates. Concordance between MT-MS and CI was observed for 1105 isolates (91.5%), while 74 isolates (6.1%) were misidentified. Molecular identification revealed that 73 of these 74 isolates were identified correctly by MT-MS and CI correctly identified the last one. Concordance between the two techniques was excellent for the medically-important species (98-100%), including the identification of closely-related species (Candida albicans/C. dubliniensis; C. inconspicua/C. norvegensis; C. parapsilosis/C. metapsilosis/C. orthopsilosis). Only 2.3% of isolates belonging to C. famata, C. lambica and C. magnoliae or to Geotrichum spp. and Trichosporon spp. were not identified by MT-MS. This investigation highlights the potential of MT-MS-based yeast identification as a reliable, time and cost-efficient alternative to CI.
Asunto(s)
Micosis/microbiología , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Levaduras/aislamiento & purificación , Costos y Análisis de Costo , ADN Espaciador Ribosómico/química , ADN Espaciador Ribosómico/genética , Francia , Humanos , Laboratorios de Hospital , Tipificación Molecular/economía , Tipificación Molecular/métodos , Técnicas de Tipificación Micológica/economía , Técnicas de Tipificación Micológica/métodos , Micosis/diagnóstico , Estudios Prospectivos , Reproducibilidad de los Resultados , Especificidad de la Especie , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/economía , Factores de Tiempo , Levaduras/clasificaciónRESUMEN
We report here that modifications of the preanalytical steps of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) identification of yeasts, with regard to the original protocol provided by the manufacturers, appear to be efficient for the reliable routine identification of clinical yeast isolates in medical laboratories. Indeed, when one colony was sampled instead of five and the protein extraction protocol was modified, the performance of MALDI-TOF MS was superior to that of the API ID 32C method (discrepancies were confirmed by using molecular identification), allowing the correct identification of 94% of the 335 clinical isolates prospectively tested. We then demonstrated that the time for which the primary cultures were preincubated on CHROMagar did not impact the identification of yeasts by MALDI-TOF MS, since 95.1 and 96.2% of the 183 clinical yeast isolates prospectively tested were correctly identified after 48 and 72 h of preincubation, respectively.
Asunto(s)
Técnicas Microbiológicas/métodos , Micología/métodos , Micosis/diagnóstico , Manejo de Especímenes/métodos , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Levaduras/química , Levaduras/clasificación , Medios de Cultivo/química , Humanos , Factores de Tiempo , Levaduras/crecimiento & desarrollo , Levaduras/aislamiento & purificaciónRESUMEN
Invasive fungal infections (IFI) have become a major public health problem in industrialized countries, notably due to the increasing number of immunocompromized individuals. Endogenous candidiasis, arising from the patient's commensal flora, accounts for the majority of IFI. C albicans, generally originating from the colonized gastrointestinal tract, is the causative agent in about 50% of cases. Molecular and cellular investigations are helping to decipher the mechanisms underlying the commensal-pathogen transition. We have found that neutropenic patients are generally colonized by a single genotype, and that all C. albicans genotypes can cause invasive infection. By using epithelial cell-based models, we have shown that alpha 1, 2 and beta 1, 2 mannosides present in the outermost layer of the fungal cell wall mediate adherence to enterocytes. In addition, C. albicans uses different strategies for epithelial cell invasion, depending on the precise cell type with which it interacts.
Asunto(s)
Candida albicans/patogenicidad , Candida albicans/genética , Enterocitos/microbiología , Células Epiteliales/microbiología , Genotipo , HumanosRESUMEN
The human pathogenic fungus Candida albicans can cause systemic infections by invading epithelial barriers to gain access to the bloodstream. One of the main reservoirs of C. albicans is the gastrointestinal tract and systemic infections predominantly originate from this niche. In this study, we used scanning electron and fluorescence microscopy, adhesion, invasion and damage assays, fungal mutants and a set of fungal and host cell inhibitors to investigate the interactions of C. albicans with oral epithelial cells and enterocytes. Our data demonstrate that adhesion, invasion and damage by C. albicans depend not only on fungal morphology and activity, but also on the epithelial cell type and the differentiation stage of the epithelial cells, indicating that epithelial cells differ in their susceptibility to the fungus. C. albicans can invade epithelial cells by induced endocytosis and/or active penetration. However, depending on the host cell faced by the fungus, these routes are exploited to a different extent. While invasion into oral cells occurs via both routes, invasion into intestinal cells occurs only via active penetration.
Asunto(s)
Candida albicans/citología , Candida albicans/fisiología , Enterocitos/citología , Enterocitos/microbiología , Células Epiteliales/citología , Células Epiteliales/microbiología , Células CACO-2 , Diferenciación Celular/fisiología , Línea Celular Tumoral , Interacciones Huésped-Patógeno , Humanos , Microscopía Electrónica de Rastreo , Microscopía FluorescenteRESUMEN
In France, children with confirmed congenital toxoplasmosis receive a treatment for a period of 12 to 24 months. Such prolonged treatment may generate potentially severe risks, in particular hematologic and cutaneous. Our objective is to compare the effectiveness of two therapeutic strategies on the prevention of retinochoroiditis by a randomized, non-inferiority, open-label, parallel study including 486 children, 3 to 6 months of age with a non-severe form of congenital toxoplasmosis. Following randomization, pyrimethamine-sulphonamide treatment is initiated for a period of three months, followed by a treatment with Fansidar(®) for 9 months, or therapeutic abstention. Follow-up visits during a two-year period will include an examination of the eye, a blood test, and questionnaires to evaluate the children's quality of life and their parents' anxiety. Confirming the non-inferiority of the effectiveness of a short-term treatment will improve the quality of life of parents and children.
Asunto(s)
Coroiditis/prevención & control , Toxoplasmosis Congénita/tratamiento farmacológico , Antiinfecciosos/uso terapéutico , Antimaláricos/uso terapéutico , Coroiditis/diagnóstico , Coroiditis/etiología , Femenino , Estudios de Seguimiento , Humanos , Lactante , Masculino , Pirimetamina/uso terapéutico , Calidad de Vida , Sulfonamidas/uso terapéutico , Toxoplasmosis Congénita/complicaciones , Toxoplasmosis Congénita/diagnóstico , Resultado del TratamientoRESUMEN
Nowadays, many commercial kits allowing the detection of digestive parasites by DNA amplification methods have been developed, including simplex PCR assays (SimpPCRa) allowing the identification of a single parasite, and multiplex PCR assays (MultPCRa) allowing the identification of several parasites at once. Thus, aimed at improving the diagnosis of intestinal protozoal infections, it is essential to evaluate the performances of these new tools. A total of 174 DNA samples collected between 2007 and 2017 were retrospectively included in this study. Performances of four commercial SimpPCRa (i.e., CerTest-VIASURETM) and three MultPCRa (i.e., CerTest-VIASURETM, FAST-TRACK-Diagnostics-FTD-Stool-ParasiteTM and DIAGENODE-Gastroenteritis/Parasite-panel-ITM) were evaluated for the detection of Cryptosporidium spp., Entamoeba spp., and Giardia intestinalis in stool samples compared to our routinely used in-house SimpPCRa. Globally, the SimpPCRa showed better sensitivity/specificity for the detection of G. intestinalis, E. histolytica, E. dispar, and Cryptosporidium spp. (i.e., 96.9/93.6%; 100/100%; 95.5/100%; and 100/99.3%, respectively), compared to the three commercial MultPCRa tested. All in all, we showed that MultPCRa offer an interesting alternative for the detection of protozoans in stool samples depending on the clinical context.
RESUMEN
A one-year prospective survey of fungal air contamination was conducted in outdoor air and inside two haematological units of a French hospital. Air was sampled with a portable Air System Impactor. During this period of survey, the mean viable fungal load was 122.1 cfu/m(3) in outdoor air samples, and 4.1 and 3.9 cfu/m(3) in samples from adult and pediatric haematology units, respectively. In outdoor samples, Cladosporium was the dominant genus (55%) while in the clinical units, Penicillium sp. (23 to 25%), Aspergillus sp. (15 to 23%) and Bjerkandera adusta (11 to 13%) were the most frequently recovered airborne fungi. The outdoor fungal load was far higher in autumn (168 cfu/m(3)), spring (110 cfu/m(3)) and summer (138 cfu/m(3)) than in winter (49 cfu/m(3)). In indoor air, fungal concentrations were significantly lower in winter (2.7 to 3.1 cfu/m(3)) than in summer (4.2 to 5.0 cfu/m(3)) in both haematology units. In the outdoor environment, Penicillium sp. and Aspergillus sp. were more abundant in winter while the levels of Cladosporium were lowest during this season. In the haematological units, the presence of Aspergillus sp. was stable during the year (close to 20%), Bjerkandera sp. was particularly abundant in winter (close to 30%); levels of Penicillium sp. were highest in autumn while levels of Cladosporium sp. were highest in spring and summer.
Asunto(s)
Microbiología del Aire , Contaminación del Aire Interior/análisis , Hongos/aislamiento & purificación , Material Particulado/análisis , Monitoreo del Ambiente , Francia , Arquitectura y Construcción de Hospitales , Estaciones del AñoRESUMEN
BACKGROUND: Genotyping studies have shown heterogeneity of Candida albicans flora in patients with human immunodeficiency virus infection, with possible co-existence of multiple clones with distinct resistance patterns. We report the result of a prospective study aimed at investigating the dynamics and heterogeneity of C. albicans flora in patients with de novo acute leukemia. DESIGN AND METHODS: Between 2001 and 2003, 66 consecutive adults with newly diagnosed acute leukemia were monitored for Candida colonization. From 19 patients with repeated multi-site C. albicans colonization, eight were randomly selected and multiple isolates from each individual mucosal site were genotyped sequentially over time using microsatellite markers. RESULTS: Despite topical use of polyenes, 60.6% of the patients were colonized repeatedly and at multiple sites. Altogether, 2,730 peripheral samples were cultured, 379 (13.9%) of which yielded yeasts. C. albicans was the most common species recovered (68%). From eight randomly selected patients colonized with C. albicans, 429 isolates were genotyped. Seven patients carried a unique genotype which was identical in all body niches and over the period of study. In one case, minor genotypic differences were observed. None of the patients shared C. albicans clones with identical genotypic profiles. Candidemia occurred in one of eight patients and the blood strain genotype did not differ from those of colonizing isolates. The genotypic profile was not altered by topical and/or systemic use of antifungal agents in any of the patients. CONCLUSIONS: In patients with de novo acute leukemia, genetic evolution of the colonizing C. albicans flora and selection of variants or replacement of the original strain upon antifungal drug pressure or nosocomial transmission are rare events.
Asunto(s)
Candida albicans/genética , Candidiasis/microbiología , Genoma Fúngico , Leucemia/complicaciones , Neutropenia/complicaciones , Enfermedad Aguda , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Antifúngicos/farmacología , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Candida albicans/efectos de los fármacos , Candidiasis/etiología , Farmacorresistencia Fúngica/genética , Femenino , Variación Genética , Genotipo , Humanos , Huésped Inmunocomprometido , Leucemia/tratamiento farmacológico , Masculino , Persona de Mediana Edad , Agonistas Mieloablativos/efectos adversos , Agonistas Mieloablativos/uso terapéutico , Neutropenia/inducido químicamente , Especificidad de Órganos , Aislamiento de Pacientes , MuestreoRESUMEN
C. albicans is a commensal yeast of the mucous membranes in healthy humans that can also cause disseminated candidiasis, mainly originating from the digestive tract, in vulnerable patients. It is necessary to understand the cellular and molecular mechanisms of the interaction of C. albicans with enterocytes to better understand the basis of commensalism and pathogenicity of the yeast and to improve the management of disseminated candidiasis. In this study, we investigated the kinetics of tight junction (TJ) formation in parallel with the invasion of C. albicans into the Caco-2 intestinal cell line. Using invasiveness assays on Caco-2 cells displaying pharmacologically altered TJ (i.e. differentiated epithelial cells treated with EGTA or patulin), we were able to demonstrate that TJ protect enterocytes against invasion of C. albicans. Moreover, treatment with a pharmacological inhibitor of endocytosis decreased invasion of the fungus into Caco-2 cells displaying altered TJ, suggesting that facilitating access of the yeast to the basolateral side of intestinal cells promotes endocytosis of C. albicans in its hyphal form. These data were supported by SEM observations of differentiated Caco-2 cells displaying altered TJ, which highlighted membrane protrusions engulfing C. albicans hyphae. We furthermore demonstrated that Als3, a hypha-specific C. albicans invasin, facilitates internalization of the fungus by active penetration and induced endocytosis by differentiated Caco-2 cells displaying altered TJ. However, our observations failed to demonstrate binding of Als3 to E-cadherin as the trigger mechanism of endocytosis of C. albicans into differentiated Caco-2 cells displaying altered TJ.
Asunto(s)
Candida albicans/fisiología , Candidiasis/metabolismo , Endocitosis , Interacciones Huésped-Patógeno , Intestinos/microbiología , Uniones Estrechas/metabolismo , Uniones Estrechas/microbiología , Células CACO-2 , Candidiasis/microbiología , Humanos , Mucosa Intestinal/metabolismo , Intestinos/ultraestructura , Uniones Estrechas/ultraestructuraRESUMEN
Members of the Fusarium group were recently detected in water distribution systems of several hospitals in the world. An epidemiological investigation was conducted over 2 years in hospital buildings in Dijon and Nancy (France) and in non-hospital buildings in Dijon. The fungi were detected only within the water distribution systems of the hospital buildings and also, but at very low concentrations, in the urban water network of Nancy. All fungi were identified as Fusarium oxysporum species complex (FOSC) and Fusarium dimerum species complex (FDSC) by sequencing part of the translation elongation factor 1-alpha (TEF-1α) gene. Very low diversity was found in each complex, suggesting the existence of a clonal population for each. Density and heterogeneous distributions according to buildings and variability over time were explained by episodic detachments of parts of the colony from biofilms in the pipes. Isolates of these waterborne populations as well as soilborne isolates were tested for their ability to grow in liquid medium in the presence of increasing concentrations of sodium hypochlorite, copper sulfate, anti-corrosion pipe coating, at various temperatures (4°-42 °C) and on agar medium with amphotericin B and voriconazole. The waterborne isolates tolerated higher sodium hypochlorite and copper sulfate concentrations and temperatures than did soilborne isolates but did not show any specific resistance to fungicides. In addition, unlike waterborne isolates, soilborne isolates did not survive in water even supplemented with glucose, while the former developed in the soil as well as soilborne isolates. We concluded the existence of homogeneous populations of FOSC and FDSC common to all contaminated hospital sites. These populations are present at very low densities in natural waters, making them difficult to detect, but they are adapted to the specific conditions offered by the complex water systems of public hospitals in Dijon and Nancy and probably other localities in the world.
Asunto(s)
Agua Potable/microbiología , Fusarium/aislamiento & purificación , Microbiología del Agua , Abastecimiento de Agua , Aclimatación , Antifúngicos/farmacología , Biopelículas , Sulfato de Cobre/farmacología , Francia , Fusarium/genética , Fusarium/crecimiento & desarrollo , Hospitales , Factor 1 de Elongación Peptídica/genética , Compuestos de Fósforo/farmacología , Dióxido de Silicio/farmacología , Hipoclorito de Sodio/farmacología , Microbiología del Suelo , TemperaturaRESUMEN
A novel strategy for the diagnosis of systemic candidosis was evaluated, based on the combination of two enzyme immunoassays that detect a candida oligomannoside repetitive epitope expressed in large amounts by Candida albicans (Platelia Candida Ag), and antibodies against C. albicans mannan, the major cell-wall immunogen in which this epitope is present (Platelia Candida Ab). Sera were selected retrospectively from intensive care and haematology patients with clinically suspected systemic candidosis, and from whom Candida spp. had been isolated from normally sterile sites. Of the 21 patients infected with C. albicans, 13 had positive antigenaemia and 14 had a positive antibody response, including eight patients who were antigenaemia negative. The sensitivity of the combined tests was 100%. In patients infected with C. glabrata (n = 12) or C. tropicalis (n = 10), the sensitivity was 83% and 80%, respectively. For the remaining patients, infected with C. parapsilosis (n = 10), C. krusei (n = 8) or C. kefyr (n = 2), the sensitivity of the combined tests was 40%, 50% and 50%, respectively. At least one of the serological tests was positive before yeast growth occurred in 60% of patients for whom a serum sample was available before blood culture sampling. An increase in serological test positivity to >80% was observed for sera obtained around the date of positive culture, irrespective of the Candida species isolated. These results suggest that regular serological monitoring for both mannanaemia and anti-mannan antibodies in at-risk patients may contribute to the early diagnosis of candidosis.
Asunto(s)
Anticuerpos Antifúngicos/sangre , Candida/patogenicidad , Candidiasis/diagnóstico , Mananos/sangre , Mananos/inmunología , Adulto , Anciano , Candida/inmunología , Candidiasis/inmunología , Niño , Estudios de Cohortes , Femenino , Humanos , Técnicas para Inmunoenzimas , Lactante , Masculino , Persona de Mediana Edad , Estudios RetrospectivosRESUMEN
The dimorphic yeast Candida albicans is a component of the normal microflora at the mucosal surfaces of healthy individuals. It possesses an array of phenotypic properties considered as virulence traits that contribute to pathogenicity of the yeast in immuno-compromised patients. We addressed the question of the pathogenicity of lineages of C. albicans with regard to their genotype in three series of C. albicans isolates (a series of commensal isolates collected in healthy individuals, a group of bloodstream isolates and a group of non-bloodstream clinical isolates) using a Multi-Locus Microsatellite Typing (MLMT) approach based on the analysis of the polymorphism of 11 microsatellite loci. The MLMT analysis of the three series, corresponding to 174 C. albicans isolates, gave a 100% typability to the method, with a DP index of 0.999. The UPGMA analysis showed that the isolates segregated in eight phylogenetic groups. Interestingly, the clustering was comparable when using NJ and MS-tree algorithms and a good concordance index of the clustering was observed with MLST. All in all our data strongly indicated MLMT as a reliable tool for DNA-typing studies in C. albicans. Isolates from healthy and non-healthy individuals segregated at the same proportions into the eight phylogenetic groups, suggesting that isolates of different origin share the same overall pathogenicity. Surprisingly allelic frequencies at the HIS3 microsatellite differed significantly in commensal isolates (group A) from pooled groups B and C (clinical isolates), raising the possibility that some individual alleles at the HIS3 microsatellite may be associated with distinct pathogenic profiles in C. albicans.
Asunto(s)
Candida albicans/clasificación , Candidiasis/microbiología , Técnicas de Tipificación Micológica/métodos , Candida albicans/genética , Candida albicans/aislamiento & purificación , Portador Sano/microbiología , Estudios de Casos y Controles , ADN de Hongos/análisis , ADN de Hongos/genética , Genes Fúngicos , Marcadores Genéticos/genética , Humanos , Repeticiones de Microsatélite , FilogeniaRESUMEN
Dijon Hospital is a French tertiary care institution undergoing major renovation, and different microbiological controls revealed the presence of Fusarium spp. in the water distribution system. Because some Fusarium spp. can cause life-threatening opportunistic infections in immunocompromised patients, an 8-month survey was conducted in two hospital sites in order to evaluate the prevalence of the fungi in the water system. In 2 units of one hospital site, 100% of the samples of tap-water were positive, with high concentrations of Fusarium spp. (up to 10(5)cfu/L). In the second hospital site, 94% of samples were positive, but generally with lower concentrations. The analysis of translation elongation factor 1α (TEF) sequences of 146 isolates revealed the presence of two different Fusarium species: F. oxysporum was detected in all units explored of both hospital sites, and F. dimerum only in one unit of one hospital site. For both species, we suggest that the fungi discovered could be particularly adapted to an aquatic environment.
Asunto(s)
Monitoreo del Ambiente , Fusarium/aislamiento & purificación , Hospitales Universitarios , Calidad del Agua , Abastecimiento de Agua , Francia , Fusarium/genética , Microbiología del AguaRESUMEN
BACKGROUND: The Dijon University Hospital in Dijon, France is involved in a large construction program with heavy truck traffic and a very dusty environment. This study aimed to assess the impact of outdoor hospital construction work on Aspergillus air contamination in the immediate environment of patients at high risk for aspergillosis in the presence of protective measures. METHODS: Prospective air and surface sampling (n=1301) was performed in 3 hospital units over a 30-month period. Generalized estimating equations were used to test the relationship between Aspergillus air contamination and the different variables (construction period, air treatment system, and surface contamination). RESULTS: Positivity rates of Aspergillus spp varied from 21.1% before construction work to 16.9% during work for air samples (P=.07), and the associated mean fungal load varied from 1.21 colony-forming units (CFU)/m(3) to 0.64 CFU/m(3) (P=.04). In multivariate analysis, only the use of an air treatment system was associated with decreased airborne Aspergillus contamination (P < .0001). No significant difference was observed between the presence or absence of construction work and the proportion of airborne Aspergillus contamination (P=.91) or the Aspergillus fungal load (P=.10). CONCLUSIONS: No influence of hospital construction work on airborne Aspergillus contamination was demonstrated when protective measures were taken, including reinforcement of the importance of environmental cleaning.
Asunto(s)
Microbiología del Aire , Aspergilosis/prevención & control , Aspergillus/aislamiento & purificación , Infección Hospitalaria/prevención & control , Arquitectura y Construcción de Hospitales , Control de Infecciones/métodos , Francia , Servicio de Limpieza en Hospital , HumanosRESUMEN
BACKGROUND: Invasive filamentous fungi infections resulting from inhalation of mold conidia pose a major threat in immunocompromised patients. The diagnosis is based on direct smears, cultural symptoms, and culturing fungi. Airborne conidia present in the laboratory environment may cause contamination of cultures, resulting in false-positive diagnosis. Baseline values of fungal contamination in a clinical mycology laboratory have not been determined to date. METHODS: A 1-year prospective survey of air and surface contamination was conducted in a clinical mycology laboratory during a period when large construction projects were being conducted in the hospital. Air was sampled with a portable air system impactor, and surfaces were sampled with contact Sabouraud agar plates. The collected data allowed the elaboration of Shewhart graphic charts. RESULTS: Mean fungal loads ranged from 2.27 to 4.36 colony forming units (cfu)/m(3) in air and from 0.61 to 1.69 cfu/plate on surfaces. CONCLUSIONS: Strict control procedures may limit the level of fungal contamination in a clinical mycology laboratory even in the context of large construction projects at the hospital site. Our data and the resulting Shewhart graphic charts provide baseline values to use when monitoring for inappropriate variations of the fungal contamination in a mycology laboratory as part of a quality assurance program. This is critical to the appropriate management of the fungal risk in hematology, cancer and transplantation patients.
Asunto(s)
Microbiología Ambiental , Hongos/aislamiento & purificación , Laboratorios de Hospital , Micología , Recuento de Colonia Microbiana , Hospitales Universitarios , Humanos , Estudios ProspectivosAsunto(s)
Dirofilariasis/diagnóstico , Hernia Inguinal/diagnóstico , Conducto Inguinal/parasitología , Anciano , Animales , Diagnóstico Diferencial , Dirofilariasis/patología , Ingle/parasitología , Ingle/patología , Hernia Inguinal/patología , Humanos , Conducto Inguinal/patología , Masculino , Zoonosis/etiología , Zoonosis/parasitologíaRESUMEN
Cryptosporidium sp. isolates from a waterborne outbreak of diarrhea in France were analyzed by PCR-restriction fragment length polymorphism analysis and sequencing of the Cpgp40/15 locus. Ninety-one percent of the isolates were Cryptosporidium hominis type Ib. The results of this study and those of studies of other outbreaks suggest that the type Ib allele is the predominant allele associated with waterborne cryptosporidiosis.
Asunto(s)
Alelos , Cryptosporidium/clasificación , Brotes de Enfermedades , Gastroenteritis/epidemiología , Proteínas Protozoarias/genética , Agua/parasitología , Animales , Criptosporidiosis/epidemiología , Criptosporidiosis/parasitología , Cryptosporidium/genética , Cryptosporidium/aislamiento & purificación , ADN Protozoario/análisis , Francia/epidemiología , Gastroenteritis/parasitología , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Proteínas Protozoarias/metabolismo , Análisis de Secuencia de ADNRESUMEN
OBJECTIVES: To define the pathogenicity of respiration-deficient mutants of Candida glabrata, which present a reduced susceptibility to azoles, that are easily induced in vitro by exposure to these drugs or to ethidium bromide and that may be selected in vivo in patients receiving fluconazole. METHODS: Two wild-type isolates of C. glabrata were compared with their respective fluconazole- or ethidium bromide-induced petite mutants, regarding the carbohydrate and protein composition of the cell wall, as well as their surface physical properties, and also their adherence abilities and virulence in mice. RESULTS: Flow cytometric analysis of cell wall carbohydrates using several fluorescent lectins showed an increased binding of mutant cells to concanavalin A compared with their parent isolates, suggesting a greater availability or an increased amount of glucose-mannose residues at the cell surface in petite mutants. Likewise, some quantitative differences between parent and mutant isolates were shown by SDS-PAGE in protein extracts from blastoconidia. Regarding the surface physical properties, no significant differences were seen in the electrophoretic mobility determined by microelectrophoresis, but the two-phase partitioning method revealed a lower cell surface hydrophobicity for petite mutants. Moreover, mutant cells exhibited significant overexpression of CgEPA1 as revealed by real-time reverse transcription-PCR, but the adherence capacities to Caco-2 cells, a human enterocyte line, were not significantly different. Finally, in agreement with their slower growth, petite mutants were less virulent than parent isolates in a murine model of systemic infection. CONCLUSION: This low virulence in mice suggests that petite mutants could be disregarded clinically although they may arise during fluconazole therapy.