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About 60% of malignant tumors of the hepatopancreatoduodenal region are represented by neoplasms of the pancreas, accounting for 10% of the total number of malignant tumors of the gastrointestinal tract. Simultaneously with the accumulation of experience, the improvement of the instruments used and the improvement of the supply of medical institutions, the number of cases of surgical treatment of pancreatic neoplasms is growing. The geography of these interventions is expanding. Accordingly, the issues of prevention and treatment of postoperative complications during these interventions, including such a dangerous complication as bleeding, are becoming increasingly important. Postoperative bleeding is diagnosed in 22% of operated patients with a given localization of a malignant neoplasm. The review analyzes modern views on the etiology, diagnosis and treatment of bleeding after resection interventions for pancreatic tumors.

Relevance. The problem of determining intraoperative blood loss in combustiology remains relevant today. With the variety of existing methods of its assessment, the issue of optimizing transfusion therapy remains debatable. Intention. To evaluate effects of the eschar excision area on the complete blood count and to compare results of computational methods for determining volumes of blood loss with actual volumes of blood exfusion. Methodology. 73 burned patients were included in the retrospective study. Early surgical excision with simultaneous autodermoplasty was performed in all patients. To achieve the study purposes, the intervention areas were compared with changes in hemoglobin, hematocrit, volume and number of red blood cells. The prospective study included 20 volunteers who donated 450 ml of the whole blood. In this group, donated volumes were compared with calculated volumes of blood loss (according to PG. Budny and G.D. Warden). The data obtained were processed via Microsoft Office Excel 2007 and IBM SPSS Statistics 20.0 using descriptive, parametric and nonparametric statistics. Results and Discussion. According to the retrospective analysis, actual areas of excision were not consistently related to the changes in complete blood count in the postoperative period. Based on the results of prospective analysis, we were able to reveal correlations between actual volumes of blood loss and changes in hemoglobin concentrations in somatically healthy volunteers (by PG. Budny). There were no similar correlations between volumes of blood loss and decreased hematocrit (by G.D. Warden). Conclusion. According to the results of the study, we couldn't confirm the effectiveness of empirical methods for determining volumes of blood loss in burned patients. The PG. Budny formula gives representative results of blood loss volumes in healthy volunteers. However, the effectiveness of this method in combustiology requires further research.

Relevance. An important component of the diagnostic process in combustiology is the collection of anamnesis. At the same time, verification of the very fact of a burn injury of the skin, as a rule, does not seem to be a difficult task even for a novice doctor. However, specialists from the I.I. Dzhanelidze Institute regularly encounter errors in the differential diagnosis of burn injuries at the prehospital stage on the part of both ambulance teams (EMS) and surgeons (traumatologists) of non-specialized medical institutions. Each such case attracts attention and takes up a significant part of the time resource of the entire staff of the inpatient department of the emergency medical service for the process of clarifying and verifying the correct diagnosis, as well as determining the further routing of such a patient.Aim of study. To study the structure of diagnostic errors at the prehospital stage of the EMS to optimize patient routing by improving the existing organizational and methodological standards.Material and methods. A retrospective analysis of the case histories of all victims who were admitted to the inpatient department of the Emergency Medical Department of the I.I. Dzhanelidze St. Petersburg Research Institute of Emergency Medicine during the period from January 2018 to December 2019.Results. 4,951 patients were admitted with a leading diagnosis of the referring institution, suggesting a history of burn injury. The incidence of diagnostic errors at the prehospital stage of emergency care was 410 cases (8.3%), while burn injury was completely excluded in 178 cases (3.6%).Conclusions. 1. The results of the analysis revealed a high incidence of diagnostic errors at the prehospital stage of emergency care (8.3%), the main reason for which is the lack of awareness of differential diagnostics within the narrow specialty (combustiology) of primary contact physicians. 2. Shown is the introduction of training practice for doctors and paramedics of emergency medical services, surgeons and traumatologists of primary care in combustiology cycles in specialized burn departments. 3.In order to ensure continuity in the process of providing medical care to patients with burns, it is necessary to create a unified database of convalescents to form a feedback channel with the outpatient clinic during the implementation of the rehabilitation complex.

Analysis of available literature and our own experience, the main causes of the lysis / rejection of autoderm grafts are systematized. It is shown that there is no consensus among combustiologists about the causes of these complications. It is believed that the leading cause of such complications is the general condition of the body, the unpreparedness of wounds for surgical treatment and the development of disseminated intravascular coagulation syndrome in seriously ill people. The frequency of unsatisfactory treatment results also correlates with the level of contamination of the burn wound. An increase in microbial seeding to 107 or 108 colony forming units per gram is a relative contraindication to surgery, while the optimal level for skin grafting is considered to be 104 colony forming units per gram. In addition, locally plastic interventions are possible with a microbial load of 105 colony forming units per gram, subject to compensation for violations of the general condition of the victim. Of great importance are the timing of autodermotransplantation. Conducting skin plastics later than 14 days after the injury increases the risk of postoperative complications. This is due to the peculiarities of the pathophysiological processes characteristic of burn wounds. During the first two weeks from the moment of the injury, the processes of reparative regeneration prevail in them, however, their intensity progressively decreases over time. It has been demonstrated that immune system disorders also have a significant effect on the rate of graft lysis. Domestic experts have developed an original method for predicting the likelihood of an unsatisfactory result of skin grafting, based on an assessment of a number of parameters of the immune system. The results of our observations indicate that the main causes of lysis / rejection of autodermotransplantants in the postoperative period are insufficient preparation of wounds for surgical treatment, changes in the general condition of the body, impaired immune system (autoaggression), and a high level of microbial contamination. The development of common criteria for assessing a patients readiness for skin grafting to prevent rejection of transplanted autoderm grafts remains an urgent task of combustiology.

The results of three-year research on the use of allogeneic mesenchymal stem cells of adipose tissue (AMSCs) in the treatment of skin burns of II-III degree are presented. in a complex with wounds dressing of nanofibers chitosan and copolyamide, hyaluronic acid. It was found that with surgical necrectomy, introduction of AMSCs and substitution of defects with natural polymer coatings, the healing time is reduced by 89% (p < 0.05). Isolated administration of MSC reduces the healing period by no more than 5% (p > 0.05). The combined use of wounds dressings of nanofibers chitosan and copo lyamide with MSC accelerates the regeneration process by 26% (p < 0.05), with the introduction of AMSCs accelerating the development of granulation tissue by the fifth day of observation by 83% (p < 0.01). Joint use of wound coverings on the basis of hyaluronic acid with AMSCs is accompanied by an increase in the number of vessels of the microcirculatory bed in the defect area by 185% (p < 0.01). Clinical evaluation of the effectiveness of drugs with stem cells – a gel for topical application and a suspension of MSC LC for injection administration demonstrate their ability to optimize regeneration in the burn zone. Application of gel with AMSCs reduces the duration of epithelialization of border (dermal) burns by 2.2-2.4 times, with the final healing period being reduced by 59% (p < 0.01) and the suppuration frequency by 30% (p < 0.05). The introduction of a suspension of AMSCs into the zone of deep burn increases the frequency of engraftment of autografts, stimulates angiogenesis and proliferation of fibroblasts in the superficial and deep layers of the dermis. In the area of MSC administration, the LC perfusion level and the amplitude of blood flow fluctuation are twice as high as the values in the zones without the introduction of cells.

Purpose. To evaluate the results of the application of allogenic fibroblasts in patients with extensive thermal burns. Methods. The study included 34 patients with burns over 20% TBSA, treated in the burn ICU. In accordance with the indications for transplantation of fibroblasts, all patients were divided into 4 groups: stimulation of regeneration of superficial burns (n = 10), stimulation of regeneration after meshed skin graft (n = 14), reduction of healing time for donor wounds (n = 6) and acceleration growth of granulations after the escharectomy (n = 4). Results. A positive effect of fibroblast transplantation was found in 7 (70%) patients of the first group, 13 (93%) of the patients in the second group, 2 (33%) of the third group and 2 (50%) in the fourth study group. Conclusion. The maximum positive effect of transplantation of human dermal fibroblasts can be observed when performing the meshed skin graft and treating the superficial burns.

To reduce the incidence of gastrointestinal bleeding in patients with advanced burns by developing a prophylactic algorithm.The study consisted of retrospective group of 488 patients with thermal burns grade II-III over 20% of body surface area and prospective group of 135 patients with a similar thermal trauma. Standard clinical and laboratory examination was applied. Instrumental survey included fibrogastroduodenoscopy, endoscopic pH-metry and invasive volumetric monitoring (PICCO plus). Statistical processing was carried out with Microsoft Office Excel 2007 and IBM SPSS 20.0.New algorithm significantly decreased incidence of gastrointestinal bleeding (p0.001) and mortality rate (p=0.006) in patients with advanced burns.Цель исследования - снизить частоту развития желудочно-кишечных кровотечений у пациентов с обширными ожогами путем применения разработанного профилактического алгоритма. Материал и методы. Исследование состояло из ретроспективной части, в которую вошли результаты лечения 488 пострадавших с термическими ожогами II-III степени на площади более 20% поверхности тела, и проспективной части, в которую включены 135 пациентов с аналогичной по тяжести термической травмой. В ходе исследования использовались стандартные клинические и лабораторные методы. В объем инструментальных исследований вошли фиброгастродуоденоскопия, эндоскопическая pH-метрия и инвазивный волюметрический мониторинг (PICCO plus). Статистическую обработку данных проводили с помощью программ Microsoft Office Excel 2007 и IBM SPSS 20.0. Вывод. Внедрение разработанного алгоритма позволило свести к минимуму частоту развития желудочно-кишечных кровотечений (p0,001) и снизить летальность (p=0,006) в группе пациентов с обширными ожогами.

International audience; Objective of the study. - Study of the spatio-temporal fungal colonization in a new medical mycology laboratory. Methods. - A 17-month survey of airborne fungal contamination was conducted in a new medical mycology laboratory at a tertiary care university hospital. This survey was implemented at three different periods: before the new premises were occupied (period A), during the move into the new laboratory (period B) and after resumption of the mycological activities in these new premises (period C). Results. - During period A, the airborne fungal load ranged from 2.3 to 6 cfu/m(3). The most frequently recovered airborne fungi were Penicillium spp. (75 to 100%). During period B, a dramatic increase in Penicillium chrysogenum conidia was observed in the air of the new laboratory (40 to 160 cfu/m(3)). During period C, the fungal load ranged from 4.5 to 8.4 cfu/m(3). Penicillium was the most common genus identified in rooms of the laboratory where no filamentous fungi were handled, while Aspergillus was clearly the predominant genus (78%) in the room dedicated to the culture of filamentous fungi. Conclusions. - We suggest that the specific fungal ecology in air of the room dedicated to the culture of filamentous fungi is due to the handling of a large number of medical strains of A. fumigatus.; Objectif de l’étude : Étude spatio-temporelle de la colonisation fongique dans un nouveau laboratoire de mycologie médicale. Méthodes : Durant 17 mois, l’aérocontamination fongique a été évaluée dans trois pièces d’un nouveau laboratoire de mycologie médicale d’un centre hospitalo-universitaire. Cette surveillance a été programmée à trois périodes différentes : avant l’occupation des nouveaux locaux (période A), durant l’emménagement dans le nouveau laboratoire (période B) et après la reprise des activités de mycologie médicale dans ces nouveaux locaux (période C). Résultats : Durant la période A, la charge fongique dans l’air était comprise entre 2,3 et 6 ufc/m3. Le micromycète le plus fréquemment isolé a été Penicillium spp. (75 à 100 %). Durant la période B, une très forte augmentation de la charge en conidies de Penicillium chrysogenum a été observée dans l’air du nouveau laboratoire (40 à 160 ufc/m3). Durant la période C, la charge fongique a été comprise entre 4,5 et 8,4 ufc/m3. Penicillium a été le genre le plus fréquemment identifié dans les deux pièces du laboratoire où aucun champignon filamenteux n’est manipulé, alors qu’Aspergillus a clairement été le genre prédominant (78 %) dans la pièce consacrée à la culture des champignons filamenteux. Conclusions : Nous suggérons que la manipulation d’un grand nombre de souches médicales d’Aspergillus est à l’origine de l’écologie spécifique qui existe dans la pièce du laboratoire destinée à la manipulation des champignons filamenteux.

L’identification conventionnelle (IC) des levures est fondee sur l’utilisation des tests morphologiques, immunologiques et biochimiques (galerie API® 32 C, bioMerieux). La spectrometrie de masse MALDI-TOF (SM) a ete proposee recemment comme nouvelle approche pour l’identification des microorganismes. L’objectif de notre etude etait de comparer prospectivement les performances analytiques de la SM et de l’IC pour l’identification des isolats cliniques de levures. En cas de discordance d’identification, le sequencage des regions ITS de l’ADN ribosomal etait utilise comme methode d’identification de reference. Un total de 1 207 souches a ete analyse. Une concordance entre SM et IC a ete observee pour 1 105 souches (91,5 %) alors que la proportion de souches differemment identifiees par IC et SM n’etait que de 6,1 % (74 souches). Parmi ces 74 identifications discordantes entre SM, l’identification par biologie moleculaire confirmait l’identification obtenue par SM pour 73 isolats (6 %) et celle obtenue par IC pour 1 isolat (0,1 %). Pour les principales especes d’interet medical, les concordances d’identification entre les deux techniques etaient excellentes (entre 98 et 100 %) y compris pour les especes phylogenetiquement proches (Candida albicans/Candida dubliniensis; Candida inconspicua/Candida norvegensis; Candida parapsilosis/Candida metapsilosis/Candida orthopsilosis). La SM n’a ete mise en defaut que pour 2,3 % des souches appar tenant aux especes Candida famata, Candida lambica, Candida magnoliae et aux genres Geotrichum sp. et Trichosporon sp. Nos investigations soulignent le potentiel de la SM pour l’identification des levures et comme alternative fiable, rapide et de moindre cout par rapport aux methodes conventionnelles.

In clinical laboratories, isolation of Candida species is generally based on the culture of specimens on Sabouraud dextrose agar. This strategy does not allow species identification on primary culture and makes it difficult to detect mixed cultures. Chromogenic media contain substrates that react specifically with different Candida species, and partly overcome these difficulties. Objectives The aim of this study was to compare two chromogenic media: (i) CandiSelect4 (C4), a new medium developed for direct identification of C. albicans and presumptive identification of C. krusei , C. tropicalis and C. glabrata (ii) CHROMagar Candida (CH), a medium licensed for direct identification of C. albicans , and presumptive identification of C. tropicalis and C. krusei , and employed in our laboratory since 2002. Materials and methods Altogether, 533 clinical specimens were seeded on both media. Identification on C4 was compared to that achieved on CH, API 32 C test or an in-house C. glabrata rapid identification test. Results Discrepant positive cultures occurred in 16 samples and were shared equally between C4 and CH. The ability of both media to detect associations was equivalent, with 34 associations identified with C4 and CH. No discrepancy was observed with respect to identification of C. albicans . C4 identified six of six C. krusei , but false-negative identification of C. tropicalis occurred in 2/11 cases and false-positive identification of C. glabrata occurred in 2/34 cases. Conclusions The overall performance of C4 and CH appeared almost identical during a two-month comparison in the clinical mycology setting.

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.

International audience; An 18-month survey of indoor fungal contamination was conducted in one haematology unit during a period of construction work. Air was sampled with a portable Air System Impactor and surfaces with contact Sabouraud plates. During this survey the mean concentration of viable fungi in air was 4.2 cfu/m(3) and that for surfaces was 1.7 cfu/plate. At the beginning of construction work, there were increases in airborne fungal spores (from 3.0 to 9.8 cfu/m(3)) in the unit, but concentrations did not exceed 10 cfu/m(3) during the 18-month period. The most frequently recovered airborne fungi were Penicillium spp. (27-38%), Aspergillus spp. (25%) and Bjerkandera adusta, a basidiomycete identified with molecular tools (7-12%). Blastomycetes accounted for more than 50% of the fungal flora on surfaces. Investigating the impact of a new air-treatment system (mobile Plasmair units), there were significant reductions in fungal contamination for the Plasmer -treated rooms, and in these rooms we observed the same level of fungal load whether construction work was in progress or not.

This study was undertaken to identify prescribing policies likely to favour or limit fluconazole resistance within a clinical department. Fluconazole exposure within the infectious diseases and clinical haematology units was investigated, and data were compared with in vitro susceptibility of Candida albicans isolates obtained in these units. Fluconazole utilization was determined by the number of fluconazole treatment-days per 100 hospitalization days (penetration index). In the infectious diseases unit, separate evaluations for low-dose fluconazole (50mg) prescribed as intermittent or prolonged treatment, and for higher-dosing schedules (fluconazole 200mg) were made. Susceptibility of C. albicans isolates was surveyed in a broth microdilution assay by measuring the inhibitory concentration 50% (IC 50 ). The penetration index (PI) for fluconazole 50mg declined from 1992 to 1977 in infectious diseases ( P = 0.0048). In the meantime, total usage of fluconazole increased, due to increased prescribing of fluconazole 200mg ( P = 0.0724). The IC 50 of C. albicans isolates tested in infectious diseases decreased between 1994 and 1996 from 7.33mg/ml to 1.64mg/ml ( P = 0.0075). In clinical haematology, declines in C. albicans IC 50 and fluconazole PI were not significant ( P = 0.35 and P = 0.07, respectively). These data suggest that prolonged or repeated exposure to low-dose fluconazole, rather than total cumulative use, was associated with fluconazole resistance in the infectious diseases unit. Moreover, restoration of a normal ecology was observed when low-dose prolonged or intermittent prescriptions were reduced.

Molecular typing studies have shown that the predominant form of reproduction of Candida albicans is clonal and that, in a majority of situations, persistent or recurrent infections are due to a unique strain. Characterization of distinct subpopulations and correlation with clinical features may thus be important to understanding the pathogenesis of candidiasis. In a clonal model, a unique polymorphic marker may identify populations with different biological properties. We therefore compared 48 bloodstream isolates and 48 nonbloodstream matched strains of C. albicans at the elongation factor 3-encoding gene (CEF3) polymorphic microsatellite locus of C. albicans . Sizing of the alleles was performed by automated capillary electrophoresis. A new, 137-bp allele was characterized, and seven nondescribed combinations were observed, resulting in 15 and 11 distinct CEF3 profiles in bloodstream and control strains, respectively. Genotypes 126-135, 130-136, and 131-131 accounted for 60.4% of both bloodstream and control strains. Four bloodstream isolates but no control strains displayed the 135-135 combination. None of the other genotypes was present at an increased frequency in bloodstream isolates. Bloodstream and nonbloodstream strains of C. albicans thus have a heterogeneous structure at the CEF3 locus, with three major and multiple minor allelic combinations.

An experience of treatment of 133 patients with severe bums was analyzed. Bleedings from the upper parts of the gastrointestinal tract were diagnosed in 16 patients in different terms since their admission to the hospital. At the moment of carrying out of the endoscopic research all bleedings were considered as taking place. Statistically significant risk factors of the development of gastroduodenal bleedings were considered to be an alcoholic intoxication at the moment of injury and insufficient fluid therapy during the pre-admission stage and young age of the patients. The antisecretory therapy showed that the detection of risk factors in question should be regarded as an indication to the reinforced regime of preventive measures for gastroduodenal injuries.

EA MERS; International audience; 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.

Resume Une methode de culture en plaques de microtitration a ete utilisee pour etudier la sensibilite au fluconazole de levures isolees de prelevements oropharynges (n = 108) ou d'hemocultures (n = 56) et comparer le profil des souches isolees chez des patients recevant ou non une chimioprophylaxie par fluconazole. Le parametre mesure, la concentration inhibitrice 50 % (CI 50), est la concentration de fluconazole reduisant de 50 % la turbidite d'une suspension de la levure etudiee apres 48 heures de culture. Les CI 50 moyennes ont pu etre calculees pour les especes fungiques suivantes : Candida albicans, 1,26 μg/ml; C. parapsilosis, 2,38 μg/ml; C. glabrata, 4,54 μg/ml; C. krusei, 12,58 μg/ml. Neuf des 108 souches de C. albicans ont presente une CI 50 superieure ou egale a 4 μg/ml, et les CI 50 moyennes des souches de C. albicans isolees chez des patients recevant ou non du fluconazole au moment du prelevement etaient de 3,11 et 0,93 μg/lm respectivement (p

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.

We developed a simple method for the identification of Candida glabrata on the basis of the ability of this species to rapidly assimilate trehalose but not sucrose. After incubation of yeasts with Rosco diagnostic tablets containing sucrose or trehalose, identification of C. glabrata was achieved in 4 h with 100% sensitivity and specificity.

International audience; 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.

International audience; 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.

La kératite amibienne est une infection rare mais grave, provoquée par un protozoaire ubiquiste du genre Acanthamoeba. Dans la majorité des cas elle est liée au port de lentilles de contact. Nous présentons une observation de kératite amibienne secondaire à un traumatisme végétal chez un patient non porteur de lentilles de contact. Cette observation souligne l’importance d’un diagnostic précoce de la kératite amibienne, qui conditionne le pronostic visuel. Le diagnostic biologique repose le plus souvent sur la mise en évidence en culture des kystes et des trophozoïtes à partir d’un grattage ou d’une biopsie cornéenne. Le traitement est long, difficile et souvent mis en échec. La prise en charge de cette infection demande donc un dialogue étroit entre clinicien et biologiste, un prélèvement de qualité et des tests de laboratoire rapides et performants.

Analysis of a polymorphic microsatellite locus was applied to 85 Candida albicans strains from healthy individuals. Comparison with strains from nonhealthy individuals previously analyzed in our laboratory showed an overall similarity, suggesting that all commensal strains have the ability to develop as pathogens.

In order to determine the specificities of PCR-based assays used for detecting Cryptosporidium parvum DNA, eight pairs of previously described PCR primers targeting six distinct regions of the Cryptosporidium genome were evaluated for the detection of C. parvum , the agent of human cryptosporidiosis, and C. muris , C. baileyi , and C. meleagridis , three Cryptosporidium species that infect birds or mammals but are not considered to be human pathogens. The four Cryptosporidium species were divided into two groups: C. parvum and C. meleagridis , which gave the same-sized fragments with all the reactions, and C. muris and C. baileyi , which gave positive results with primer pairs targeting the 18S rRNA gene only. In addition to being genetically similar at each of the eight loci analyzed by DNA amplification, C. parvum and C. meleagridis couldn’t be differentiated even after restriction enzyme digestion of the PCR products obtained from three of the target genes. This study indicates that caution should be exercised in the interpretation of data from water sample analysis performed by these methods, since a positive result does not necessarily reflect a contamination by the human pathogen C. parvum.

We developed a PCR-based method that can be used to identify Cryptosporidium parvum in human feces. Fecal oocysts were concentrated by centrifugation on a sodium chloride gradient and filtration on a nitrocellulose filter prior to DNA extraction and PCR amplification of a 452-bp C. parvum-specific DNA sequence with a protocol including dUTP and uracil-N-glycosylase. All samples obtained from naturally infected humans (n = 10), calves (n = 4), and goats (n = 2) were positive. A 100% detection rate was achieved with both formed and solid stools (n = 10) seeded with 1,000 C. parvum oocysts per g. Procedures based on stool concentration by a modified Ritchie method and subsequent oocyst identification by immunofluorescent labeling or acid-fast staining require concentrations of 50,000 to 500,000 oocysts per g to achieve a 100% detection rate with formed stools. The described PCR-based assay thus has a 50- to 500-fold increase in sensitivity compared to those of the methods commonly used to analyze formed feces.

PURPOSE: In patients with neutropenia, thoracic computed tomography (CT) halo and air-crescent signs are recognized as major indicators of invasive pulmonary aspergillosis (IPA). Nevertheless, the exact timing of CT images is not well known. PATIENTS AND METHODS: Seventy-one thoracic CT scans were analyzed in 25 patients with neutropenia with surgically proven IPA. RESULTS: On the first day of IPA diagnosis with early CT scan (d0), a typical CT halo sign was observed in 24 of 25 patients. At that time, the median number of thoracic lesions was two (range, one to six), and pulmonary involvement was bilateral in 12 cases. The halo sign was present in 68%, 22%, and 19% of cases on d3, d7, and d14, respectively. Similarly, the air-crescent sign was seen in 8%, 28%, and 63% of cases on the same days. Otherwise, a nonspecific air-space consolidation aspect was seen in 31%, 50%, and 18% of cases on the same days. The analysis of calculated aspergillary volumes on CT showed that, despite antifungal treatment, the median volume of lesions increased four-fold from d0 to d7, whereas it remained stable from d7 to d14. Overall, 21 patients (84%) were cured by the medical-surgical approach. CONCLUSION: In patients with neutropenia, CT halo sign is a highly effective modality for IPA diagnosis. The duration of the halo sign is short, and it demonstrates the value of early CT. The increase of the aspergillosis size on CT in the first days after IPA diagnosis is not correlated with a pejorative immediate outcome when using a combined medical-surgical approach.