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R Grillot

Publications and source records attributed to R Grillot.

At least 19 recordsLinked to original sources
PubMed2002

Eight-year surveillance of environmental fungal contamination in hospital operating rooms and haematological units.

An eight-year fungal environmental surveillance was carried out in 15 operating theatres and two haematological units. Sampling was performed twice a year in each room, using contact plates for plane surfaces and sterile swabs for grids. From 1992 to 1999, individual rooms in the 17 units were sampled on 1094 occasions and 3822 samples were collected. The percentage of rooms without fungus increased regularly between 1992 and 1999 (41.1% and 74.8%, respectively). The units were classified according to the fungal contamination during the eight years: the operating theatres which required the highest protection (cardiological, thoracic, vascular, hand, orthopaedic and neurosurgery) and the adult haematological unit showed least contamination (71.8% rooms were negative). The most frequent species isolated were Penicillium spp. (28.4%), Cladosporium spp. (15.6%) and Aspergillus spp. (7.6%). Aspergillus fumigatus was rarely isolated (3.7%), and was mainly isolated at the beginning of the study. This study demonstrates that environmental control programmes are effective in reducing environmental mould contamination and could be useful in establishing exposure guidelines, especially by defining an acceptable level of biocontamination in zones at risk.

Aspergillus↗
PubMed2002

Effect of medium composition on static and cidal activity of amphotericin B, itraconazole, voriconazole, posaconazole and terbinafine against Aspergillus fumigatus: a multicenter study.

The effect of the medium composition on the fungistatic (MIC) and fungicidal (MLC) activity of amphotericin B, itraconazole, voriconazole, posaconazole and terbinafine against four Aspergillus fumigatus strains has been investigated by four European laboratories. MICs were determined by broth microdilution, using RPMI 1640 and Antibiotic Medium 3 (AM3), three times in three independent determinations by the four laboratories. MLCs were determined for the three independent determinations by the four laboratories, subculturing 100 microl from each well showing no visible growth after 48 hours. Except for a 2-dilution difference observed in three cases, no differences were observed between MICs determined on the two media. In contrast, a 3- to 6-dilution discrepancy between the MLCs was observed for the azoles. Endpoints on RPMI were higher than those on AM3. A 1-2 dilution difference was noted between both the endpoints of amphotericin B and of terbinafine. The highest inter- and intra-laboratory agreements were reached on AM3. The azoles showed a medium-dependent fungicidal activity.

Amphotericin B↗
PubMed2001

A longitudinal study of lung transplant recipients infected with Aspergillus: genetic polymorphism of A fumigatus.

BACKGROUND: Aspergillus infection is a well-known complication of lung transplantation and remains associated with high mortality rates. Molecular typing methods are required to elucidate the complex epidemiology of Aspergillus disease in lung transplant recipients. METHODS: Eight lung transplant recipients from one hospital were followed for A fumigatus colonization or infection. Forty-four sequential isolates from these patients were selected and typed by three molecular methods (random amplified polymorphic DNA, sequence-specific DNA primer and multi-locus enzyme electrophoresis). RESULTS: Sixteen different types were identified of which 14 were specific to 1 patient. A factorial correspondence analysis showed that variability between sequential isolates from a single patient was as high as between isolates from the other patients. Lung transplant recipients presented many different genotypes, reflecting the environmental diversity of A fumigatus. Nevertheless, throughout their follow-up, 2 of the 8 lung transplant recipients harbored a common genotype that was not replaced by others. CONCLUSIONS: These results confirm the important genetic polymorphism of the A fumigatus population. The observed genotypes were not related to the type of Aspergillus disease or anti-fungal treatment used nor to the outcome of the patient. These data confirm that all A fumigatus molecular types present the same pathogenic risk.

Adult↗
PubMed2001

Evaluation of Candida ID, a new chromogenic medium for fungal isolation and preliminary identification of some yeast species.

Candida ID, a new chromogenic medium, allows identification of Candida albicans (blue colonies) and preliminary identification into a group of four species (pink colonies). In comparison with Albicans ID2 and Sabouraud gentamicin chloramphenicol on 446 fungal strains, Candida ID allowed the isolation of more species than Albicans ID 2 (95.5% versus 91.2%).

Chromogenic Compounds↗
PubMed2001

Genetic polymorphism of Aspergillus fumigatus in clinical samples from patients with invasive aspergillosis: investigation using multiple typing methods.

The genotypes of 52 strains of Aspergillus fumigatus isolated from 12 patients with invasive aspergillosis were investigated using three typing methods (random amplified polymorphic DNA, sequence-specific DNA polymorphism, and microsatellite polymorphism) combined with multilocus enzyme electrophoresis. Isolates were from patients hospitalized in three different geographic areas (Lyon, France; Grenoble, France; and Milan, Italy). In each case, the genetic polymorphism of several colonies (two to five) within the first respiratory clinical sample was studied. For the 52 isolates tested, random amplified polymorphic DNA identified 8 different genotypes, sequence-specific DNA polymorphism identified 9 different types, and microsatellite polymorphism identified 14 types. A combination of these results with multilocus enzyme electrophoresis study identified 25 different types within the sample studied. We identified 3 patients (of the 12 studied) who carried a single genotype; 6 patients were infected by two genotypes, 1 patient had four genotypes, while the last patient had five. A combination of typing methods provided better discrimination than the use of a single method. Typing methods revealed a population structure within each geographical site, suggesting that the epidemiology of A. fumigatus should be considered separately for each of these geographic areas. This study demonstrates the usefulness of combining several typing methods in reaching an understanding of the epidemiology of A. fumigatus and clarifies whether it is sufficient to type one isolate from each specimen to determine the strain involved in invasive aspergillosis.

Aspergillosis↗
PubMed2001

Hydrocortisone, prednisolone and dexamethasone act on Aspergillus fumigatus in vitro susceptibility to itraconazole.

In a previous in vitro investigation from the same laboratory a therapeutic level of hydrocortisone enhanced the itraconazole susceptibility of a single strain of Aspergillus fumigatus. In the present work, the influence of therapeutic levels of hydrocortisone (1 microM), prednisolone (0.125 microM 0.25 microM and 0.5 microM) and dexamethasone (0.25 microM and 0.5 microM) on the itraconazole susceptibility of four A. fumigatus strains, was determined. A. fumigatus conidia were germinated either in the absence or in the presence of a glucocorticoid. The germinated conidia were then spread onto plates and grown either in the presence or in the absence of a glucocorticoid, together with increasing concentrations of itraconazole. The mean colony forming units (CFU) were measured. Two factor analyses of variance showed that hydrocortisone significantly (p <0.001) potentiated the action of itraconazole. The cytotoxic effect of prednisolone on the fungal strains added significantly to the effect of itraconazole (p <0.001). Dexamethasone was also cytotoxic to the fungus but, when used in conjunction with itraconazole, it effectively increased (p <0.01) the number of CFU. This study showed a direct effect of glucocorticoids, currently in use for patient therapy, on in vitro A. fumigatus susceptibility to itraconazole.

Antifungal Agents↗
PubMed2000

Emerging pathogens.

The ever increasing numbers of immunosuppressed individuals has led to a significant increase in the incidence of opportunistic infections, particularly those caused by fungi. The epidemiology of infections caused by the common fungal pathogens such as Candida albicans, Cryptococcus neoformans and Aspergillus fumigatus has been well documented. However, in addition to these, a number of species which have previously been unrecognized (e.g., C. dubliniensis) or have previously been assumed to be non-pathogenic (e.g., Saccharomyces cerevisiae, Scedosporium spp. and Fusarium spp.) have emerged as agents of human disease. Since these species have only been identified recently as human pathogens, their role in disease is poorly understood. In most cases, identification of these species is problematic and therefore their epidemiology has yet to be elucidated adequately. In addition, several of these species fail to respond to conventional antifungal therapies. In this article, we describe the emergence of two separate yeast species (C. dubliniensis and S. cerevisiae) and two separate groups of moulds (Scedosporium prolificans and Fusarium spp.), as human pathogens. It is apparent from what we already know, that much work has yet to be performed before we have a clear understanding of how these species cause disease and most importantly how they can be controlled.

Candida↗
PubMed2000

Aspergillus Fumigatus antigen detection in sera from patients at risk for invasive aspergillosis.

We have developed an inhibition enzyme immunoassay (inhibition-EIA) to monitor for the occurrence of invasive aspergillosis (IA) in sera from 45 immunocompromised (IC) patients. The test uses rabbit polyclonal antibodies and a mixture of components from Aspergillus fumigatus, containing three predominant antigens with molecular weights of 18,000, 33,000, and 56,000. Circulating antigens were found in five of seven proven cases of IA due to A. fumigatus. In two of the five positive cases, antigenemia was detected with inhibition-EIA earlier than with X ray or other biological methods. No antigens were detected in the sera from two patients with proven IA due to Aspergillus flavus and Aspergillus terreus nor in the sera from four patients with probable IA. Circulating antigens were not detected in the control group, composed of 30 healthy adult blood donors. Four of the 32 at-risk patients examined, though they displayed no definite evidence of IA, gave a positive result in this test. The sensitivity, specificity, and positive predictive value of inhibition-EIA were 71.4, 94.4, and 71.2%, respectively. The data were compared with those obtained by a latex agglutination assay of galactomannan (GM) that was positive in only one patient with probable IA. The higher sensitivity obtained by inhibition-EIA may well be due to its ability to detect circulating antigens other than GM in the sera of IC patients with IA. Detecting these antigens may improve the diagnosis of IA, as they may serve as markers of this infection.

Antigens, Fungal↗
PubMed1999

Combination of three typing methods for the molecular epidemiology of Aspergillus fumigatus infections. European Research Group on Biotype and Genotype of Aspergillus.

This study investigated the source of infection and strain relatedness of Aspergillus fumigatus isolates from bronchial colonisation and invasive aspergillosis (IA) in four transplant patients. Environmental isolates from the patient's home and from the hospital and infecting isolates were obtained for patient A who developed IA. Clinic environmental and colonising isolates were obtained for patient B. Sequential isolates were obtained from various organs from patient C who developed IA and also from patient D who had a bronchitic aspergillosis that developed into IA. Ninety-one A. fumigatus isolates were analysed by three typing methods: multi-locus enzyme electrophoresis (MLEE), random amplified polymorphic DNA (RAPD) and sequence-specific DNA primers (SSDP). The three combined typing methods demonstrated a greater differentiation of isolates than the typing methods used separately or in pairs. This demonstrated the genotypic variability of A. fumigatus and facilitated better epidemiological analysis. Large polymorphisms were demonstrated for each patient isolate between and colonies within various samples. The relatedness of the isolates suggested nosocomially acquired aspergillosis for patient B, but the source of infection for patient A remained unclear. The results suggested at least three multiple infections among the four patients. This study enabled the identification of the source of infection and strain relatedness, which in turn facilitates the development of preventive measures for patient management in the future.

Aspergillosis↗
PubMed1999

[Epidemiology of fungemia in a university hospital; therapeutic incidence].

Twenty-two candidemia happened in our hospital from January 1997 to may 1998. We studied the clinical evolution of the patients and the sensitivity of the yeasts to antifungal therapy (Fungitest and E-Test method). We found 11 Candida albicans (CA), 10 Candida non albicans (CNA) (3 C. glabrata, 2 C. parapsilosis, 4 C. tropicalis, 1 C. krusei) and 1 Saccharomyces cerevisiae. The mean age of the patients was 56.4 years. There were 13 men and 9 women. We found one group of 8 (36.4%) oncohematological patients, one group of 8 (36.4%) patients with abdominal surgery, one group of 3 (13.6%) children and one group of 3 adults (13.6%) who spent more than 10 days in an intensive care unit. Ten times, these candidemia were associated with bacteriemia, 4 times with several bacteria. Three patients died because of the candidemia, 2 times with CNA and one time with CA. There wasn't any resistance to amphotericin B or ketoconazole. All the CA and 3 CNA (30%) remained sensitive to the four antifungal drugs we used (amphotericin B, ketoconazole, fluconazole, itraconazole). The 3 C. glabrata and the C. krusei were resistant or limit to fluconazole. Since the generalization of the use of fluconazole, the epidemiology is marked by the emergence of new strains of CA with high level of resistance to azols, and of CNA. In our hospital, the CA remain preponderant and only the CNA are resistant to fluconazole making difficult the choice of empiric treatment for serious fungemia.

Adult↗
PubMed1998

Use of simulated blood cultures to compare a specific fungal medium with a standard microorganism medium for yeast detection.

The aim of this study was to compare the performance of a specific blood culture medium recently developed to detect fungi on the Bactec 9240 system (Mycosis IC/F; Becton Dickinson Diagnostic Instrument Systems, USA) with that of the standard medium used for detection of bacteria (Aerobic Plus/F; Becton Dickinson). Simulated blood cultures were performed on 43 strains belonging to ten fungal species using the two media. The mean time to yeast detection using the Mycosis IC/F medium was 29.03+/-13.99 h, in contrast to a mean time of 73.92+/-56.74 h using the Aerobic Plus/F medium. These results show that the Mycosis IC/F medium clearly reduces the time to yeast detection on the Bactec 9240.

Automation↗
PubMed1998

A monoclonal antibody specific to surface antigen on Candida krusei.

A monoclonal antibody (MAb; MAb 6B3) which reacts specifically with a cell wall antigen found in all strains or isolates of Candida krusei was developed. MAb 6B3 was extensively tested by immunofluorescence assay for cross-reaction with many Candida, Cryptococcus, Saccharomyces, Trichosporon, and Rhodotorula species and was found to react only with the species C. krusei. The specific epitope is expressed on the surface of fungal cells and appears to reside on a protein moiety. Taking into account the increasing importance of fluconazole-resistant strains in nosocomial fungal infections, the very high degree of specificity of this MAb for C. krusei could be useful for the routine detection of C. krusei in culture or in tissue samples.

Animals↗
PubMed1998

[Fungal and parasitic nosocomial infections: importance and limitations of disinfection methods].

Due to the increase of opportunistic mycosis and parasitosis for several years, the management of fungal and parasitic risk in hospital has become a necessity and an obligation. The authors remind the main rules and knowledges essential to an optimal management of the fungal and parasitic disinfection in hospital. They summarize the efficiency of different disinfection processes in relation to yeasts, filamentous fungi, Pneumocystis carinii, Cryptosporidia and Microsporidia involved in hospital pathology.

Animals↗
PubMed1998

Does hydrocortisone modify the in vitro susceptibility of Aspergillus fumigatus to itraconazole and amphotericin B?

To analyse if hydrocortisone could modify the in vitro susceptibility of Aspergillus fumigatus to antifungal drugs, we developed a procedure to test the susceptibility of an A. fumigatus strain to amphotericin B and itraconazole, grown in the presence and in the absence of hydrocortisone. Conidia were germinated in the presence or the absence of hydrocortisone in Czapek medium without antifungal drug. A dilution of these cultures (5x10(3) conidia ml(-1)) was spread onto Czapek-agarose plates containing both antifungal drug and hydrocortisone. The cfu per plate were enumerated and compared. A therapeutic concentration of hydrocortisone induced a significant increase in the susceptibility to itraconazole. Conversely, the susceptibility to amphotericin B was not significantly modified when this antifungal drug was associated with hydrocortisone.

Amphotericin B↗
PubMed1998

Aspergillus and lung transplant recipients: a mycologic and molecular epidemiologic study.

BACKGROUND: After lung transplantation, filamentous fungi and more particularly Aspergillus fumigatus are commonly isolated, although the origin of contamination is unclear. METHODS: To investigate the fungal flora in bronchoscopic fluids, we retrospectively reviewed 20 cases of lung transplant recipients. Using sequence-specific DNA primers analysis, we typed the clinical strains of A. fumigatus isolated from 6 lung transplant recipients. For 4 of them, the strains of this species were isolated from their environment. RESULTS: At least once 90% of patients had filamentous fungi, and A. fumigatus was the most frequently isolated. Bronchial colonization was detected in 14 patients, invasive bronchial mycosis was diagnosed in 4 others, and no case of invasive pulmonary fungal infection was detected. Genome typing of the 47 clinical strains revealed that a given patient could be affected by several different strains. A very extensive polymorphism existed among the 38 environmental strains. Origin of contamination at home was possible in 1 case and in the hospital in 3 cases. CONCLUSIONS: Bronchial colonization is frequent after lung transplantation. Although the clinical strains show a polymorphism, it is less widespread than the polymorphism of environmental strains. The origin of acquisition may be in the patient's community.

Adult↗
PubMed1997

Molecular typing of Aspergillus fumigatus strains by sequence-specific DNA primer (SSDP) analysis.

A PCR typing method has been developed and tested to investigate the polymorphism of clinical strains of Aspergillus fumigatus. Firstly, the DNA fragments from random amplified polymorphic DNA (RAPD) patterns of nine epidemiologically and geographically non-related monosporal strains of A. fumigatus were cloned and sequenced. The pairs of five sequence-specific DNA primers (SSDP), characteristic of the 5' and 3' extremities of the RAPD products, were then used in high stringency PCR to type 43 clinical strains of A. fumigatus from 13 patients, according to the presence or absence of a single amplified band. This original approach, which uses the advantages of PCR, has made it possible to overcome the difficulties resulting from the low stringency amplification. The SSDP analysis of 51 A. fumigatus strains (9 unrelated monosporal strains and 43 clinical strains from 13 patients) can be classed into 22 different types with a high reproducibility and a high level of discrimination (D = 0.96). The results suggest that seven lung transplant patients with necrotizing aspergillosis, bronchitis aspergillosis and bronchial colonization were infected by multiple strain genotypes, whereas three patients with invasive aspergillosis seem to have been infected by a single strain.

Aspergillosis↗