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[Quantitative immunoelectrophoretic methods applied to the immunochemistry of Candida].

Cytoplasmic soluble extracts of yeasts were compared by quantitative immunoelectrophoretic methods. Comparative studies of Candida albicans and Candida tropicalis permitted to confirm the antigenic complexity of fungi and the existence of important cross-reacting antigens between this two yeast, to demonstrate the specificity of the Candida tropicalis cell-wall antiserum and the presence of soluble cell-wall constituents in cytoplasmic yeast extracts. Crossed-line immunoelectrophoresis with and without absorption of antibodies in situ demonstrated four specific antigens unique to Candida albicans group A cytoplasmic extract not shared with the group B. The other procedures used were not sufficient for identification, since the resulting patterns were confusingly complex.

Animals↗

[The quantitative immunochemical determination of specific antibodies against Candida albicans within the scope of immunological testing in the malignant melanoma (author's transl)].

Precipitating antibodies against a polysaccharide antigen from Candida albicans were in the human serum determined quantitatively by the aid of a reversed radial immunodiffusion technique. The gel, therewith, contained the antigens, whilst into the gel's punching holes the serum specimens and the calibrated standards were filled in. Amplifying examinations of the course in patients suffering from malignant melanoma, the findings of the methods used hitherto were compared within the scope of an immunological status (in vivo-skin testing by dinitrochlorobenzene, Candida antigen, trichophytin, and tuberculin, precipitation test after Ouchterlony, immunofluorescence-optical evidence of Candida antibodies as well as determination of the Candida antibodies. In contrast to the hitherto available qualitative or semi-quantitative methods evidencing humoral antibodies, we consider the feasibility of simply workable quantitative determinations of antibodies to be an enrichment of the scale of methods. The reversed Mancini technique, simultaneously, is appropriate to determine specific antibodies against other diagnostically relevant antigens, so far as precipitating antigen/antibody complexes are formed. Therewith, commercially available testing kits could favour the standardization.

Antibodies, Fungal↗

[Antimycotic sensitivities of the clinically isolated candidas].

82 candida strains isolated from clinical material were tested for their sensitivity to nystatin, 5-fluorocytosine (5-FC) and to clotrimazole. Strains were showing high M.I.C. values against nystatin and 45% primary resistance to 5-FC by the tube dilution technique. No resistance was noted to clotrimazole. The resistance, developed in candida strains against several antifungal drugs, will establish the problem encountered in the antibiotic treatment of bacterial infections. Also the necessity of the sensitivity tests with the isolated candida strains in candida infections is emphasized.

Antifungal Agents↗

Frequency of Candida sp infection in tuberculous patients with acquired immunodeficiency syndrome: morphological and immunocytochemical study in sputum.

Sputum samples from AIDS patients with and without pulmonary tuberculosis were analyzed morphologically and immunocytochemically to determine the frequency of occurrence of Candida sp. Mycobacterial infection was detected by bacterioscopy and/or culture and cytological evaluation was performed using Papanicolaou and Toluidine Blue staining. Immunoreaction for Candida sp was performed using polyclonal antibody in selected cases with fungal structures in smears stained by the Papanicolaou or Toluidine Blue method. An increased frequency of Candida sp (2.5 times) was observed in the tuberculous group compared to the group of AIDS patients without tuberculosis. The Toluidine Blue stain showed good results for the detection of Candida sp in sputum. Due to the increased risk of this opportunistic infection among more severely immunocompromised patients. Toluidine Blue staining of sputum samples submitted to analysis seems to be a reliable screening method.

AIDS-Related Opportunistic Infections↗

Person-to-person transfer of Candida albicans in the spacecraft environment.

We assessed the exchange of Candida albicans among crew members during 10 Space Shuttle missions. Throat, nasal, urine and faecal specimens were collected from 61 crew members twice before and once after space flights ranging from 7 to 10 days in duration; crews consisted of groups of five, six or seven men and women. Candida albicans was isolated at least once from 20 of the 61 subjects (33%). Candida strains were identified by restriction-fragment length polymorphism (RFLP) after digestion by the endonucleases EcoRI and HinfI; further discrimination was gained by Southern blot hybridization with the C. albicans repeat fragment 27A. Eighteen of the 20 Candida-positive crew members carried different strains of C. albicans in the specimens collected. Possible transfer of C. albicans between members of the same crew was demonstrated only once in the 10 missions studied. We conclude that the transfer of C. albicans among crew members during Space Shuttle flights is less frequent than had been predicted from earlier reports.

Candida albicans↗

[Identification of yeasts of the Candida genus with a growth inhibition system: Microring YT].

BACKGROUND: It has been observed an spectacular increasing of opportunistic Candida yeast infections. Many of them are fatal, and rapid and effective identification of the infecting species contributes to start the correct treatment. Several new methods for yeast identification have become available; Microring YT is one of these methods based on the growth inhibition by 6 different chemical products. The aim of this work is to study the performance of the test using representative clinical yeast isolates. METHODS: A total of 146 strains belonging to the 5 most common Candida species isolated in the clinical laboratory were identified using conventional methods (germ tube and chlamydospores production, and the standard API 20C AUX and 16 sugars auxonography; Institute Pasteur) and the Microring YT System. This test uses the differing susceptibilities of yeast to 6 discs mounted on a filter paper ring. The chemical products and dyes are: janus green, ethidium bromide, triphenyl tetrazolium chloride, brilliant green, cycloheximide and rhodamine 6G. The inhibition pattern of a 6 digit code is compared with a list of profiles. RESULTS: Using the Microring YT system 112 of the 146 studied strains were correctly identified with an overall concordance of 77% between this method and the standard one. The morphological study (germ tube production) increased 6% the identification of Candida albicans. Better results were obtained with C. krusei and C. parapsilosis (85% of concordance). With C. glabrata only 59% of concordance was found. CONCLUSIONS: In spite Microring YT is a simple method, easy to perform and read, it was considered inadequate for the identification of Candida species as a routine microbiological procedure.

Bacteriological Techniques↗

[Adherence of isolated Candida albicans strains from HIV+ infected patients to buccal epithelial cells: correlation with serotype, sensitivity to fluconazole and virulence].

During infection by human immunodeficiency virus (HIV), oral pseudomembranous candidiasis frequency is high. Adherence of Candida albicans to epithelial cells constitutes one of the first step in the setting up of candidiasis. Adherence ability of Candida albicans to buccal epithelial cells (BEC) is estimated for 83 isolates. The yeasts are isolated from buccal swabs from HIV+ patients with clinical oropharyngeal candidiasis. Results show that Candida albicans ability to adhere to BEC is variable within isolates; it appears that most of the isolates are weakly adherent. In addition, adherence is not correlated with the serotype A or B of yeasts. We do not either observe any correlation between adherence capacity and sensibility of yeasts to fluconazole. Moreover, no correlation is observed between a group of 13 genetically identical isolates (analysed on 21 loci with isoenzymes method) and their adherence to BEC. Finally, Candida albicans virulence, which is studied in vivo on intravenously infected mouse model, does not seem to be correlated with capacity of each yeast to adhere on BEC in vitro.

AIDS-Related Opportunistic Infections↗

Clinical application of counterimmunoelectrophoresis in detection of Candida serum precipitins.

Precipitin antibodies to candida antigens were found by counterimmunoelectrophoresis (CIE) and double immunodiffusion (DID) in 100 per cent of sera from patients with significant candida infection. Such antibodies were also found by CIE in 63 per cent of individuals with candida colonization, 47 per cent of bacteremic patients, and 14 per cent of normal control subjects. No sera from these patients and normal control subjects were positive by DID. Serum precipitin titers by CIE in patients with significant candidiasis were significantly higher than in patients of the other groups. Detection of candida serum precipitins by CIE, when precipitin titers are obtained, compares favorably with DID as a diagnostic tool in significant candidiasis.

Antigens↗

[Hemagglutinating activity of Candida albicans culture extracts].

3M KC1 extracts of Candida albicans cultures display a haemoagglutinating activity on human and sheep red blood cells. The haemoagglutinins, may be lectins in nature, behave like antigens being fully inhibited by anti-Candida immune serum. The latter lyses human red blood cells sensitized with 3M KC1 Candida albicans extracts. Human and animal red blood cells react against Candida albicans hypertonic salt solution extracts, in double diffusion agar tests, and give one or more precipitation lines. The interpretation of the results is discussed.

Animals↗

Candida zeylanoides causing candidiasis as tinea cruris.

Candida zeylanoides caused candidiasis as tinea cruris in 4 cases. Candida zeylanoides were characterized by good growth of the strain in malt medium, potato dextrose agar and Sabourard's agar. Their colony was milky white and greasy with smooth surface. The diameter of the colony reaches 12 mm within a week. As candida type pseudohyphae but not hyphae, they do not produce ascospore and bolospore. Carbon sources fermentation showed positive results. Nitrogen sources assimilation agar (KNO3) negative, arbutin agar negative, nonvitamin medium positive. G+C mol% value of their DNA was 51.24. Animal experiment showed that guinea pigs were infected by Candida zeylanoides. The main feature of candidiasis as tinea cruris caused by the strain was pink infiltrative erythema with clear margin, a lot of grain-sized papules, a few vesicles, and thin scales. The disease can be cured by preparation of ketoconazole.

Adult↗

The in vitro lysozyme susceptibility of Candida species cultured in sucrose supplemented media.

The in vitro sensitivity to lysozyme of twelve isolates of Candida (three isolates each of C. albicans, C. tropicalis, C. glabrata and C. krusei) pre-incubated in sucrose supplemented media, was determined by a growth inhibition assay. In general, C. albicans and C. tropicalis exhibited increased susceptibility to lysozyme as the sucrose concentration in the growth medium was reduced. Candida krusei demonstrated the reverse effect and C. glabrata isolates showed different trends. Although generally the Candida species examined were susceptible to lysozyme in the decreasing order of C. glabrata, C. albicans, C. tropicalis and C. krusei, differences in susceptibility were noted among different isolates within a given species. These results imply that salivary lysozyme and dietary carbohydrates may exert selective pressure on oral colonization by Candida species.

Candida↗

[Candida albicans spondylitis: successful treatment with fluconazole. 2 case reports].

Two patients suffering from candida albicans spondylitis in the lumbar spine are reported. Both had been operated on because of malignancy. One patient's clinical course was complicated by candida septicemia, and later on, serious candida endophthalmia developed. Conventional X-rays, combined with technetium scintigraphy, showed inflammatory destruction of the end plates of neighbouring vertebrae. Computerized tomography was done in the second patient, when an abscess was detected on both sides in the psoas muscle. Consecutively, operative removal and a spondylodesis were performed. Candida albicans could be isolated from removed discal material. Treatment by fluconazole for six months, in the first case in the beginning also by amphotericin B, lead to the healing of fungal spondylitis, accompanied by partial osseous consolidation.

Aged↗

Antifungal susceptibility pattern of non-albicans Candida species & distribution of species isolated from Candidaemia cases over a 5 year period.

A marked increase in the incidence of patients of candidaemia is reported from a tertiary care hospital in north India over the last five years (15 patients in 1991 and 275 in 1995). The distribution of Candida species isolated from January 1991 through December 1995 was investigated. Antifungal susceptibility against amphotericin B, 5-fluorocytosine, ketoconazole and fluconazole of 100 randomly selected non-albicans Candida species isolated during 1995 was determined by an in-house standardized disc diffusion method and the standard broth dilution procedure recommended by the National Committee for Clinical Laboratory Standards (NCCLS, USA). The disc diffusion method correlated well with NCCLS method except for ketoconazole. Resistance against any antifungal was confirmed only by results of NCCLS method. A shift to higher isolation of non-albicans Candida species was observed during this period (52.6% in 1992 to 89.5% in 1995). Resistance was observed against 5-fluorocytosine in 3 per cent strains of C. krusei; and against fluconazole in 24.2 per cent of C. krusei, 15.4 per cent of C. guilliermondii and 5.7 per cent strains of C. tropicalis. No resistance was detected against amphotericin B and ketoconazole. Thus Candida species with fluconazole resistance have become more prominent in recent years.

Antifungal Agents↗

Diarrhoea associated with Candida spp.: incidence and seasonal variation.

To study the incidence and seasonal variation of diarrhoea associated with Candida, 978 diarrhoeal stool specimens from patients of all age groups were examined by microscopy and culture. Candida spp. was the sole pathogen (unassociated with other diarrhoeagenic bacteria, protozoa, or helminths) in 15.3% of the total cases. Candida albicans (94.9%) was the predominant species isolated. The incidence was highest among the infants aged 0-12 months (37.1%), followed by a decline in the rest of the children aged less than 5 years with a second peak in the people aged over 5 years, including adults. The paediatric age group had a significantly higher incidence in the summer season compared to the rainy (p < 0.05) or winter (p < 0.01) months. Thus, there was a definite age and seasonal variation in the incidence of diarrhoea caused by the overgrowth of Candida in the Varanasi region of the Indian subcontinent.

Adolescent↗

In vitro susceptibility of 115 isolates of Candida to amphotericin B, fluconazole and itraconazole.

Opportunistic fungal infections are an increasingly important cause of morbidity and mortality particularly due to Candida species (1). There is also an increase of candidosis especially ascribed to acquired or induced immunodeficiency syndromes or in the event of long-term antibiotic, immuno-suppressor or cytotoxic therapies. Consequently there has been an increase in the use of systemic antifungal agents responsible for the emergence of new opportunistic fungi (2) and resistant species (3, 4). Oropharyngeal candidiasis caused by various species of Candida is one of the most common opportunistic infections in AIDS. In recent years there has been an increasing number of yeast isolates resistant to fluconazole (4, 5) or to amphotericin B (6). The aim of the present study was to examine the susceptibility in vitro of itraconazole, a newly introduced antifungal agent in the local or systemic therapy of oropharyngeal candidiosis, vs well-known agents such as amphotericin B and fluconazole, against various Candida clinical isolates. The present results, in agreement with other studies, show strong in vitro activity of itraconazole against Candida spp. and particularly against less susceptible species C. glabrata, C. tropicalis or C. krusei.

Amphotericin B↗

[Chronic eosinophilic pneumonia associated with allergy to candida and with changes in bronchoalveolar lavage fluid].

A 61-year-old woman was admitted to the hospital with the chief complaints of fever, coughing, and shortness of breath. A chest X-ray film revealed infiltrates in both upper lung fields. A specimen was obtained by transbronchial lung biopsy and chronic eosinophilic pneumonia was diagnosed. The patient' s condition improved after the start of corticosteroid therapy. A relapse occurred, and corticosteroid therapy was again successful. Because an environmental provocation test was positive, allergological examinations were done. An intradermal test for sensitivity to candida was positive, as was a test for precipitation of antibodies to candida. In addition, an inhalation provocation test with candida antigen was positive. Therefore, allergy to candida was believed to have caused this patient's symptoms. Bronchoalveolar lavage was done three times. Eosinophils predominated in the first sample, but lymphocytes predominated in the last two. This raises the possibility that chronic eosinophilic pneumonia and bronchiolitis obliterans organizing pneumonia might change into each other.

Bronchoalveolar Lavage Fluid↗

[In vitro susceptibility to antifungal agents of Candida strains isolated from patients with various diseases of the respiratory tract].

The aim of the study was the estimation of in vitro susceptibility to antifungal agents of yeast isolated from sputum of 70 respiratory diseases patients using the disc-diffusion method-antimycogram. The following agents were tested: amphotericin B, 5-fluorocytosine, nystatin, ketoconazole, fluconazole. Only Candida strains were isolated from sputum, 82% of them were Candida albicans. We noted differences in susceptibility to antimycotics of Candida strains. The best antimycotic in vitro was 5-fluorocytosine. 54% of isolated Candida strains were resistant to 1 or more antimycotics.

Adult↗

[A review of infectious endocarditis due to Candida].

OBJECTIVE: As fungal endocarditis is a serious disease, frequently requiring cardiac surgery, a review was made of the experience of our Departments in this pathology. DESIGN: A retrospective analysis of clinical, echocardiographic and surgical data. SETTING: Patients studied in a tertiary care Hospital with cardiac surgery available. PATIENTS: Between 1984 and 1994 there were ten cases of candida endocarditis in nine patients, four male and five female, mean age--45 +/- 12 years (31-65). INTERVENTIONS: The following parameters were analysed: clinical (predisposing factors, clinical evolution, complications, therapy and mortality), echocardiographic (presence of vegetations, abscesses, valvular regurgitations). Patients studied in other Centres and referred to our Department only for examination (echocardiograms) were excluded from this analysis. RESULTS: Eight cases in seven patients were prosthetic valve endocarditis and two native valve endocarditis. No patient was drug addicted. Seven cases of prosthetic valve endocarditis developed less than one year after surgery and another had a gynecological fungal infection as the cause of the endocarditis. Four patients had had previous endocarditis. There were four embolic events and three developed heart failure. There were three perivalvular infections, six valvular regurgitations and only one case with huge vegetations on echocardiography. Nine patients were treated with amphotericin B, in five fluocytosin was added and in four ketoconazol, which was replaced by flukonazol in one patient. Therapy was continued for at least eight weeks. Six patients were operated during the acute stage and one died. One patient was operated on late after the infection. Three patients died during the active stage. In a follow up of 5.2 +/- 4.8 years (8 months to 8 years) there was one fatal candida endocarditis relapse, one fatal candida sepsis, one non cardiac death, one patient developed a periprosthetic leak and one had recurrent systemic embolization. Abscesses/pseudoaneurysms were found in five out of seven patients submitted to surgery. CONCLUSION: Candida infective endocarditis has a bad prognosis, specially in those patients not operated early; it develops in patients with predisposing factors, which in our series were a previous infective endocarditis (four patients) and/or a prosthetic valve implantation less than one year before; it has important morbidity with multiple embolic events, perivalvular involvement, valvular regurgitation and heart failure.

Adult↗