Positive peritoneal fluid fungal cultures in postoperative peritonitis after bariatric surgery.

Postoperative peritonitis (POP) is a common surgical complication after bariatric surgery (BS). We assessed the importance of positive fungal cultures in these cases of POP admitted to the intensive care unit. Clinical features and outcome were compared in 25 (41%) Candida-positive patients (6 (22%) fluconazole-resistant Candida glabrata) and 36 patients without Candida infection. Candida infections were more commonly isolated in late-onset peritonitis and were often associated with multidrug-resistant bacteria. Risk factors for intensive care unit mortality (19.6%) were diabetes and superobesity. Candida infections, including fluconazole-resistant strains, are common in POP after BS. These data encourage the empirical use of a broad-spectrum antifungal agent.

Pneumocystosis: a network survey in the Paris area 2003-2008.

The aims of this network group were to collect epidemiological data of PcP cases in 14 hospitals in the Paris area and to determine the Di-Hydro Pteroate Synthase (DHPS) genotypes, genetic markers for possible sulfamide resistance. From January 1, 2003 to December 31, 2008, 993 (mean 166/year) PcP cases have been reported. Sixty-five percent of patients were HIV-positive. The median count of CD4 lymphocytes was 32/mm(3) (30 in HIV-positive patients, 152 in HIV-negative patients). In HIV-positive patients, PcP revealed the HIV infection in 39%. Among 304 PcP occurring in HIV known infected patients, no prophylaxis was prescribed for 64%; cotrimoxazole prophylaxis had been prescribed to 47 patients but only one of them had the right compliance. In HIV-negative patients (264), corticosteroids were prescribed in 59% and cytotoxic chemotherapies in 34%; 78% did not receive prophylaxis. One hundred sixty nine tumoral pathologies and 116 transplantations were notified. The mortality rate was 16% at day 14 (13% in HIV-positive patients, 26% in HIV-negative patients). Mutations in DHPS genes were detected in 18.5% of samples; 12.5% of patients were infected with several strains. The total annual number of cases has been stable for five years but the proportion of HIV-negative patients increased from 25% to 43%.

Comparison of antifungal MICs for yeasts obtained using the EUCAST method in a reference laboratory and the Etest in nine different hospital laboratories.

In routine laboratory practice, the determination of MICs of antifungals for yeasts often relies on the Etest, because of a good correlation with reference methods. However, this correlation was established through predesigned studies, rather than prospective testing. The surveillance programme of fungaemia (YEASTS programme), implemented since 2003, facilitated our comparison of the Etest and the EUCAST results, obtained on a routine basis in nine different hospitals and in a reference laboratory, respectively. The analysis included 690 isolates recovered from blood culture (362 Candida albicans, 113 Candida glabrata, 69 Candida parapsilosis, 55 Candida tropicalis, 31 Cryptococcus neoformans, and 60 other yeast species) that were tested for their susceptibility to amphotericin B (n = 655), fluconazole (n = 669), itraconazole (n = 198), voriconazole (n = 588), flucytosine (n = 314), and caspofungin (n = 244). Agreement between the Etest and EUCAST datasets was calculated and categorized on the basis of previously published breakpoints. The level of agreement at ±2 dilutions was 75% for amphotericin B and 90% for flucytosine; for the azoles, it ranged from 71% for itraconazole to 87% for voriconazole. No significant difference was observed among the yeast species, except for Cryptococcus neoformans and flucytosine, with an agreement <40%. Categorical agreement ranged from 60% for itraconazole to 90% for flucytosine. Major and very major discrepancies occurred in <12% and 6%, respectively. The Etest, even when performed on a routine basis, shows a ≥71% agreement with the EUCAST reference method.

[Invasive pulmonary aspergillosis in AIDS]. / Aspergilloses pulmonaires invasives au cours du SIDA.

Risk factor for invasive pulmonary aspergillosis in HIV-negative patients include neutropenia, corticosteroid therapy, and chemotherapy. Corresponding risk factors in HIV-positive patients have not yet been reported. A case-control study was conducted at the Bichat-Claude Bernard Teaching Hospital, Paris, France, between 1991 and 1996. Eight cases were identified. In three cases, the diagnosis was documented histologically. Of the remaining five patients, four had a de novo lung cavity with a positive bronchoscopy sample, and one had a pulmonary infiltrate with a positive bronchoscopy sample in the absence of any other potential pathogen. Each case was matched with three controls who were admitted during the same period and had CD4 counts lower than 50/mm3. Median age was 38.1 years in the cases and 38.4 years in the controls. Median CD4 counts were 12.5 +/- 19.2 in the cases versus 19.3 +/- 16.3 in the controls (P = 0.14). No case-control differences were found for AIDS duration, neutrophil counts at diagnosis or during the previous six months, history of corticosteroid therapy or chemotherapy, or number of previous opportunistic infections. Cases were more likely than controls to have a preexisting lung cavity (3/8 versus 0/24; P = 0.01) and had spent more time in the hospital during the previous year (7 +/- 4.5 versus 2.8 +/- 3.2 weeks; P = 0.02). These data do not support a role for neutropenia or immunosuppressive treatments as risk factors for invasive aspergillosis in AIDS. They suggest that AIDS patients with a lung cavity or frequent hospital stays are at increased risk for invasive aspergillosis.

Cryptococcus neoformans infection in France: epidemiologic features of and early prognostic parameters for 76 patients who were infected with human immunodeficiency virus.

We retrospectively carried out a descriptive and prognostic study of 76 human immunodeficiency virus-infected patients with cryptococcosis diagnosed by a positive culture of cerebrospinal fluid (CSF), blood, urine, or other body fluid or tissue. We focused on the 65 patients with cryptococcal meningitis. At diagnosis, the mean CD4 lymphocyte count was 46/mm3; 86% of patients had fever; 67%, headache; 37%, stiff neck; 29%, altered mentation or confusion; 20% cranial nerve deficiency; and 48%, other focal deficiencies. Analysis of CSF specimens revealed the following results: normal (25% of the specimens), leukocyte count of < 20/mm3 (62%), positive India ink smear (87%), and positive cryptococcal antigen (92%). Twenty patients died within the first 3 months (3-month survival rate, 70%). A Cox regression model selected the following as prognostic parameters: age older than 30 years (relative risk [RR] = 2.1), CSF glucose level of < 2 mmol/L (RR = 3.7), previous admission to an intensive care unit (RR = 4.7), and mechanical ventilation (RR = 4.6). The outcome of cryptococcal meningitis in patients with AIDS remains difficult to predict at admission, and every case should be considered as potentially severe.

Response to fluconazole by 23 patients with human immunodeficiency virus infection and oral candidiasis: pharmacological and mycological factors.

The MICs of fluconazole for strains of Candida species and the levels of fluconazole in serum were determined at day 0 and day 14 for 23 human immunodeficiency virus-infected patients with oral candidiasis who were treated orally with 100 mg of fluconazole per day for 14 days. Among the 23 patients, 11 (48%) were not clinically cured and had persistent isolation of Candidiasis albicans (n = 10) and/or presence of non-C. albicans (n = 6). Clinical response could be predicted by the susceptibility of the strain to fluconazole determined at day 0. All 12 patients who were cured were infected with a strain for which the MIC was < 0.78 mg/liter. All four patients who were infected with a strain for which the MIC was > 3.12 mg/liter experienced clinical failure. These data suggest that a C. albicans strain could be defined as being susceptible when the MIC of fluconazole is < 0.78 mg/liter and as being resistant when the MIC is > 3.12 mg/liter.

Intestinal lipid metabolism in suckling rats infected with Giardia duodenalis.

We carried out a quantitative and qualitative analysis of intestinal digestion of neutral lipids in suckling rats infected with Giardia duodenalis. Total lipids were measured after extraction from the contents of the stomach, proximal and distal small bowel, caecum and colon. Amounts gradually fell from the stomach to the colon and were identical in infected animals and controls, although high values were occasionally found in the caecum of infected rats. Relative glyceride quantities were determined by means of high-performance thin-layer chromatography. Triglycerides were absent from the distal small bowel, and only free fatty acids and cholesterol were present in the caecum, reflecting normal digestion of neutral lipids in infected suckling rats. Our results suggest that G. duodenalis does not impair intestinal fat digestion in suckling rats.

[Frequency of bronchopulmonary isolation of Aspergillus species in patients infected with human immunodeficiency virus: pathogenic role?]. / Fréquence de l'isolement broncho-pulmonaire d'Aspergillus species chez des patients infectés par le virus de l'immunodéficience humaine: rôle pathogène?

Prevalence of aspergillosis in patients infected with the human immunodeficiency virus (HIV) is unknown. Mycologic findings in bronchopulmonary specimens from 614 HIV-positive patients sent to a parasitology laboratory between June 1, 1989 and May 31, 1991 were analyzed retrospectively; medical records of Aspergillus sp--positive patients were studied to evaluate the potential pathogenic role of this organism. Prevalence of Aspergillus sp. was 2.7% in bronchoalveolar lavage (BAL) specimens, (21/757), 15.1% in sputum specimens (3/53), 12.6% in bronchial aspirates (11/87), 7% in protected brush specimens (2/28), and 16.6% in lung biopsy specimens (4/24). A total of 20 patients (rate = 3.14%) had lung specimens positive for Aspergillus sp. (A. fumigatus 95% and A. niger 5%). Among them, 66% had a single positive specimen (14.3% with a positive smear) and 33% had several positive specimens (mean 4.9 +/- 3.3). Four patients (20%) with a more than 20-month history of AIDS and less than 36 CD4+ cells per microliter had documented pulmonary aspergillosis; three of these patients also had a current or past history of pulmonary mycobacterial infection. Rate of recovery of Aspergillus sp. in LAB specimens was low. The other specimens were harvested because of clinically suggestive manifestations. However, in 20% of patients with positive specimens. A. fumigatus was the cause of patent infection; these patients had severe AIDS-related immunodeficiency.

Detection of occult blood in stools: comparison of three gaiac tests and a latex agglutination test.

The detection of occult blood in the stools is the only simple screening method for colorectal cancer. The aim of this study was to compare the results obtained with the new Hemolex kit (Orion diagnostica, Fumouze France) with those given by three gaiac tests--Hemoccult (Smithkline diagnostics), Hemofec (Boehringer Mannheim) and Hemopreuve (Fumouze) of 165 stools from patients without special diet. Seventy-one patients with at least two positive gaiac tests or a positive Hemolex test underwent colonoscopy followed, if negative, by fibroscopy: 28 had lesions of the lower digestive tract and five of the upper digestive tract. Sensitivity, specificity and negative and positive predictive value were of 70, 98, 91 and 92% respectively for Hemolex; 82, 74, 94 and 44% for Hemoccult; 94, 67, 98 and 42% for Hemopreuve and 91, 73, 97 and 46% for Hemofec. The results obtained in this study confirm the value of the Hemolex test for the detection of human occult blood in the stools whereas the gaiac tests used are influenced by dietary components (unless restricted), explaining their poor positive predictive value. In conclusion, due to their good negative predictive values, the authors recommend that screening for colorectal tumours should be based on the use of two or three gaiac tests which should be confirmed, when positive, by an immunological test for human hemoglobin.

[Giardia intestinalis: transmission electron microscopy study of in vitro excystation]. / Giardia intestinalis: étude en microscopie électronique à transmission du dékystement in vitro.

The in vitro excystation process in Giardia intestinalis was studied by transmission electron microscopy (TEM). Untreated cysts served as controls. The excystation process was monitored by examination of organisms after the in vitro induction and at several times during the incubation phase. The control cyts had a thick wall, made of microfibrils, that appeared not to contain any weak areas. The peritrophic space extended between the cyst wall and the organism peripherally, the space was delimited by a thin cytoplasmic layer, "the outer cytoplasmic envelope" that subtended the cyst wall. During the in vitro incubation, the trophozoite cytoplasm retracted from the wall; thus, the peritrophic space became progressively larger. The outer cytoplasmic envelope detached from the cyst wall, then broke up forming numerous small vesicles lodged between the wall and the organism. The tight arrangement of the wall microfibirils was lost. Electron-dense vacuoles appeared in the peripheral cytoplasm of the trophozoite. The organism emerged through the posterior end of the cyst, leaving behind the empty husk. Emergence was followed by cell division. The possible interrelationships of biochemical and mechanical factors affecting the process of excystation are discussed in light of the present TEM findings.

Role of cytoskeleton and surface lectins in Giardia duodenalis attachment to Caco2 cells.

An in vitro model of Giardia duodenalis and the Caco2 cell line enable the study of parameters that could play a part in trophozoite attachment. We explored the role of membranous lectins of G. duodenalis in attachment-inhibition studies using carbohydrates in solution. Attachment rates were reduced by 14% and 23% in the presence of 100 mmol/l mannose-6-phosphate and glucose, respectively, as compared with control values. No significant modification was observed after trophozoite trypsinization at room temperature or at 37 degrees C. The inhibitory effects of colchicine (35%) and nocodazole (70%) suggest a primordial role of the cytoskeleton; microtubules appear to be the principal effectors of trophozoite fixation. Scanning electron microscopy revealed circular imprints on the Caco2 brush border after trophozoite detachment. The mechanisms of attachment of G. duodenalis to intestinal enterocyte-like cells in culture are thus essentially of the mechanical or hydrodynamic type; surface lectins would appear to intervene in the specificity for duodenal cells.

Adherence and multiplication of Giardia intestinalis on human enterocyte-like differentiated cells in vitro.

This report describes a technique for studying the adherence and growth of Giardia intestinalis trophozoites (strains PARIS/86/LCF/1, PARIS/86/LCF/2 and PARIS/88/LCF/8) using the human colon carcinoma cell line Caco2. Giardia trophozoites were cultured with Caco2 cells in a modified HSP3 culture medium. The biochemical differentiation of Caco2 cells was established by an increase in sucrase isomaltase activities to values of 4.51 +/- 0.90 and 10.39 +/- 3.00 milliunits/mg protein for 8- and 12-day-old cultures, respectively. Giardia, adherence to 8- and 12-day-old Caco2 cells reached a value of greater than 75% after 60 and 30 min, respectively. Adherence diminished significantly at 24 degrees C and was almost undetectable at 4 degrees C. Depletion of divalent cations reduced the proportion of adherent trophozoites by up to 46%. Adherence was pH-independent between pH 6.0 and 7.6. Parasite growth increased when Caco2 cell monolayers were used instead of axenic cultures. This in vitro human cell model may contribute to the study of the mechanisms and factors involved in the host-parasite interaction.

[Giardia intestinalis: study of infestation procedures of young mice before weaning]. / Giardia intestinalis: étude des modalités de l'infestation du souriceau avant sevrage.

Despite the prevalence of giardiasis little is known about the host-parasite specificity. With Giardia intestinalis-suckling mouse model, the success of infestation was depending on the parasite form, cyst or trophozoite, and, for the same parasite form, on the studied strain. Furthermore, the intestinal modification during the wean were unfavourable to Giardia intestinalis colonization.

In vitro excystation of Giardia from humans: a scanning electron microscopy study.

The in vitro excystation of Giardia lamblia on cysts isolated from human feces was studied. After purification by sucrose gradient, cysts were incubated in a pepsin-acid solution, then placed in a modified HSP3 medium where excystation occurred within a few minutes. The excystation procedure was studied by continuous observations by light microscopy and sequential observations by scanning electron microscopy (SEM). The in vitro excystation was stopped at timed intervals during incubation by addition of a large amount of 1% glutaraldehyde. The excystation process began by the cyst wall opening at one pole. Flagella protruded rapidly, the parasite emerged progressively from the cyst envelope, posterior end first, the empty cyst collapsed and shrank. Although flagella emerging from the organism were distinguishable, the cell body had not yet shown all the morphological features of the G. lamblia trophozoite. A radical rearrangement of the organism occurred gradually: initially oval in shape, the parasite became round, then elongated, flattened, and underwent cytokinesis. The daughter trophozoites acquired their typical morphological features: the shape, the adhesive disc with the C-shaped structure distinctly visible on the ventral surface, and the definite placement of the flagella. These observations obtained on G. lamblia by SEM were comparable to those obtained with G. muris.

[Giardia intestinalis: influence of the composition of the osmolarity of the medium on in vitro excystation]. / Giardia intestinalis: influence de la composition et de l'osmolarité du milieu sur le dékystement in vitro.

The in vitro excystation of Giardia intestinalis was studied to make the osmolarity (from 50 to 500 mosmol/l) and the components of growth medium (MCI saline solution, MCII glucose solution, MCIII nutritive solution) varying. The percentage of excystation, the viability and the generation time were determined. Excystation was observed in the saline solution between 100 to 450 mosmol/l after cyst acid pepsin incubation. The trophozoite viability was increased by glucose addition (60 min in MCI; 300 min in MCII). Only a rich medium (MCIII) permitted a generation time from 225 to 425 mosmol/l.

[Giardia intestinalis: comparative study of lipid and nitrogen fecal excretions in adults and children with parasites]. / Giardia intestinalis: étude comparee des débits fécaux lipidique et azoté chez des adultes et des enfants parasités.

10,000 faeces samples-from 9,120 adults and 880 children were examined to evaluate the faecal excretions. Giardia intestinalis was identified in 111 of those samples (84 adults and 27 children). A higher infection rate of G. Intestinalis was observed in child and male adult groups. Nitrogen excretion was evaluated and compared with fat excretion for the first time in the case of giardiasis. When malabsorption was obvious, both fat and nitrogen increased with generally moderate values. The malabsorption was much more frequent among children (88.9%) than among adults (26%) (p less than 0.001). These results could explain the fast settlement and the frequency of growth troubles in childhood. Giardiasis should be systematically and carefully investigated in a malabsorption in the person of a child.

[Immunological diagnosis of echinococcosis. A study of 146 cases, including 8 with bone involvement (author's transl)]. / Diagnostic immunologique de l'échinococcose. Etude de 146 cas dont 8 localisations osseuses.

After a study about 146 cases of echinococcosis which 8 osseous localizations, indirect immunofluorescence reactions and immunoprecipitation reactions allow for comparing between right immunologic answers in hydatid cyst of liver and hydatid cyst of bone. The right immunogenicity of osseous hydatid cyst can be explained by a close contact host-parasite and by a good vascularization and a good fertility.