Trends in the molecular epidemiology and population genetics of emerging Sporothrix species.

Sporothrix (Ophiostomatales) comprises species that are pathogenic to humans and other mammals as well as environmental fungi. Developments in molecular phylogeny have changed our perceptions about the epidemiology, host-association, and virulence of Sporothrix. The classical agent of sporotrichosis, Sporothrix schenckii, now comprises several species nested in a clinical clade with S. brasiliensis, S. globosa, and S. luriei. To gain a more precise view of outbreaks dynamics, structure, and origin of genetic variation within and among populations of Sporothrix, we applied three sets of discriminatory AFLP markers (#3 EcoRI-GA/MseI-TT, #5 EcoRI-GA/MseI-AG, and #6 EcoRI-TA/MseI-AA) and mating-type analysis to a large collection of human, animal and environmental isolates spanning the major endemic areas. A total of 451 polymorphic loci were amplified in vitro from 188 samples, and revealed high polymorphism information content (PIC = 0.1765-0.2253), marker index (MI = 0.0001-0.0002), effective multiplex ratio (E = 15.1720-23.5591), resolving power (Rp = 26.1075-40.2795), discriminating power (D = 0.9766-0.9879), expected heterozygosity (H = 0.1957-0.2588), and mean heterozygosity (Havp  = 0.000007-0.000009), demonstrating the effectiveness of AFLP markers to speciate Sporothrix. Analysis using the program structure indicated three genetic clusters matching S. brasiliensis (population 1), S. schenckii (population 2), and S. globosa (population 3), with the presence of patterns of admixture amongst all populations. AMOVA revealed highly structured clusters (PhiPT = 0.458-0.484, P < 0.0001), with roughly equivalent genetic variability within (46-48 %) and between (52-54 %) populations. Heterothallism was the exclusive mating strategy, and the distributions of MAT1-1 or MAT1-2 idiomorphs were not significantly skewed (1:1 ratio) for S. schenckii (χ2 = 2.522; P = 0.1122), supporting random mating. In contrast, skewed distributions were found for S. globosa (χ2 = 9.529; P = 0.0020) with a predominance of MAT1-1 isolates, and regional differences were highlighted for S. brasiliensis with the overwhelming occurrence of MAT1-2 in Rio de Janeiro (χ2 = 14.222; P = 0.0002) and Pernambuco (χ2 = 7.364; P = 0.0067), in comparison to a higher prevalence of MAT1-1 in the Rio Grande do Sul (χ2 = 7.364; P = 0.0067). Epidemiological trends reveal the geographic expansion of cat-transmitted sporotrichosis due to S. brasiliensis via founder effect. These data support Rio de Janeiro as the centre of origin that has led to the spread of this disease to other regions in Brazil. Our ability to reconstruct the source, spread, and evolution of the ongoing outbreaks from molecular data provides high-quality information for decision-making aimed at mitigating the progression of the disease. Other uses include surveillance, rapid diagnosis, case connectivity, and guiding access to appropriate antifungal treatment.

Clinical and microbiological characteristics of paracoccidioidomycosis in patients with AIDS in Buenos Aires, Argentina.

Paracoccidioidomycosis (Pm) is a systemic disease, endemic in the American continent. There are two different clinical forms, the infant-juvenile or subacute form (PmS) and the chronic adult form (PmC). The human immunodeficiency virus (HIV) associated paracoccidioidomycosis (PmHIV) shares characteristics with both of the previously mentioned forms. The objective of this work was to describe the epidemiological, clinical and laboratory features of the PmHIV and to compare them with the ones of PmS and the PmC. A retrospective analysis of 119 patients with paracoccidioidomycosis was performed. Ninety four suffered the chronic form, 11 the subacute one and 14 were coinfected with HIV. Patients with PmHIV presented a CD4+ T lymphocytes median of 70.5 cells/µl, 71.4% had fever, 64.3% had a miliary pattern on the chest radiography, 64.3% had hepatosplenomegaly, 64.3% had mucosal lesions and 50% had skin lesions. One patient died during his hospitalization. The clinical presentation of Pm in patients with HIV resembled the subacute form with fever, hepatomegaly and skin lesions. However, they also tended to present mucosal lesions, positive serology for Pm and pulmonary parenchyma lesions as usually seen in PmC (9/14 PmHIV patients had overlapping features, while 4/14 PmHIV patients clinically resembled PmS and 1/14 PmC). The incidence of Pm has not changed with the burden of AIDS as it has happened with other fungal infections but it appears clinically different from the classic clinical forms of the disease.

[Histoplasmosis outbreak in Morón, Buenos Aires Province, Argentina]. / Brote de histoplasmosis en la Escuela de Cadetes de la Base Aérea de Morón, Provincia de Buenos Aires, República Argentina.

A histoplasmosis outbreak affecting 6 previously healthy Air Force cadets is herein presented. The patients suffered from fever and respiratory symptoms after having cleaned an abandoned hangar soiled with pigeons and bat droppings. They all presented fever, myalgia, tachypnea, and nonproductive cough. Chest X-ray and CT scan studies showed disseminated reticulonodular images affecting both lungs. Hiliar adenomegalies were also observed. All patients achieved a favourable outcome without antifungal treatment. Both serologic tests searching for specificic antibodies (immunodiffusion and counterimmunoelectrophoresis) and histoplasmin skin tests were positive in all cases. Five soil samples mixed with pigeons and bat droppings were collected from the hangar. Suspensions of these samples were inoculated into 20 hamsters by intraperitoneal injection; mycelial phase of H. capsulatum was isolated from liver and spleen cultures. The genetic profile of this strain was compared with 12 isolates obtained from Argentinean patients, and a great degree of homogeneity was observed (> 96% similarity). Although histoplasmosis is endemic in the wet Pampas, this is the first epidemic outbreak reported south of the 34th parallel.

[Coccidioidomycosis in Argentina, 1892-2009]. / La coccidioidomicosis en Argentina, 1892-2009.

Clinical cases of coccidioidomycosis are rare in Argentina and are generally found in the large arid precordilleran area of the country. This study aims to perform a retrospective review of all coccidioidomycosis cases documented in the country from 1892 to 2009, and to describe those occurring in the last 4 years. One hundred and twenty eight cases were documented in the 117 year-period. Since the original description of the disease in 1892 until 1939, only 6 cases were registered; between 1940 and 1999, 59 (6-14/10 yrs) and the remaining 63 (49% of total cases) occurred in the last decade. The median age of 34 patients registered in 2006-2009 was 31 years (range: 7-89), male/female ratio was 1.3:1 and 12 patients were immunocompromised. Twenty-six cases were confirmed by direct microscopy and/or culture whereas the remaining ones by serology. All isolates were identified as Coccidioides posadasii. Thirty patients lived in a vast geographic region with epicenter in Catamarca Valley. Between 2006 and 2009, annual disease incidence rates in Catamarca Province increased from historical values below 0.5/100,000 to 2/100,000 inhabitants. Such increase suggests an emergency of coccidioidomycosis in that region.

[Historical evolution of some clinical and epidemiological knowledge of coccidioidomycosis in the Americas]. / Evolución de los conocimientos sobre aspectos clínico-epidemiológicos de la coccidioidomycosis en las Américas.

Coccidioidomycosis is a systemic endemic mycosis caused by two dimorphic fungi of the Coccidioides genus: Coccidioides immitis and Coccidioides posadasii. This fungal infection is only endemic in the American Continent. The majority of the epidemiological, pathogenic, clinical, mycological and therapeutical findings were obtained in the U.S.A. Coccidioidomycosis was discovered in Argentina, at the end of the XIXth century by Alejandro Posadas. In the last two decades, a new endemic zone was found in the northeast of Brazil. Several countries of the region such as Mexico, Guatemala, Honduras, Venezuela and Argentina have performed epidemiological studies which allowed a better knowledge of the endemic areas and of the clinical characteristics of this mycosis.

[Clinical and microbiological study of mycetomas at the Muñiz hospital of Buenos Aires between 1989 and 2004]. / Estudio clínico y microbiológico de los micetomas observados en el hospital de infecciosas Francisco J. Muñiz en el período 1989-2004.

This work presents clinical, microbiological and outcome data collected from 76 patients with mycetomas at the Muñiz Hospital from 1989 to 2004. Forty-nine patients were male and 27 female; the mean age was 43.4 years. The majority of the patients acquired the infection in Argentina: the most affected provinces were Santiago del Estero with 31 cases, and Chaco with 11; 8 cases came from other countries (Bolivia 6 and Paraguay 2). The mean evolution of the disease was 9.2 years. The most frequently observed sites were: feet 63 cases, ankles 3, and knees 2. Forty-eight patients had bone lesions and 5, adenomegalies. The following etiological agents were identified: Madurella grisea 29 cases, Actinomadura madurae 26, Scedosporium apiospermum 5, Nocardia brasiliensis 5, Acremonium spp. 4 (Acremonium falciforme 2, Acremonium kiliense 1, Acremonium recifei 1), Madurella mycetomatis 3, Fusarium solani 2, Nocardia asteroides 1 and Streptomyces somaliensis 1. The main drugs used in the treatments were ketoconazole and itraconazole for maduromycotic mycetomas, and cotrimoxazole associated with ciprofloxacin or amikacin for actinomycetic mycetoma. Six patients had to undergo amputation, 25 cases achieved complete clinical remission and 34 showed remarkable improvement.

Itraconazole: pharmacokinetics and indications.

Itraconazole is a highly lipophilic triazolic compound, scarcely soluble in acidified polyethylene glycol, and soluble in hydroxypropyl-beta-cyclodextrin. It possesses an excellent digestive adsorption and its peak plasma level after oral administration of 100 mg is 0.16 microgram/ml at 3 or 4 h after drug intake. Half-life of itraconazole ranges between 17 to 21 h and 99.8% binds to plasmatic proteins, especially albumin. Metabolization is mainly done in the liver where inactive metabolites are formed with the exception of hydroxy-itraconazole, which exhibits a discrete antifungal activity. Stabilization of blood levels with repeated drug administration is reached at day 14, showing an increase both in plasma concentrations and in its half-life. Tissue levels of itraconazole are 3- to 20-fold higher than plasmatic concentrations, whereas only negligible concentrations are in CSF and urine. In the skin and particularly nails, itraconazole persists for a long time after discontinuation of therapy. Its mechanism of action is similar to other azolic compounds, inhibiting the alpha-14-demethylase of lanosterol which interferes with the synthesis of ergosterol. This drug behaves as a wide spectrum antifungal agent, acting against most pathogenic fungi with the exception of the Zygomycetes. Daily doses vary, according to indications, from 100 to 400 mg. The efficacy and results obtained in dermatomycosis, candidiasis, paracoccidioidomycosis, keratomycosis, sporotrichosis, chromoblastomycosis, coccidioidomycosis, blastomycosis, cryptococcosis, phaeohyphomycosis and maduromycotic mycetomas are detailed.

Study of some lymphocyte subset counts and cytokine levels in cryptococcosis associated with AIDS.

Approximately 100 new cases per year of cryptococcosis in HIV+ patients are observed in Muñiz Hospital, 35% of them suffer a fatal outcome within the first four weeks after diagnosis in spite of treatment. Apparently there is not a useful parameter that allows a clear prediction of this early fatal outcome of the disease. The aim of this study is to determine some cytokine levels and several lymphocyte subpopulations counts in order to correlate these results with the evolution of the disease. Forty HIV+ patients suffering culture confirmed cryptococcosis were enrolled in this study, 8 HIV+ patients without cryptococcosis and 8 healthy individuals with negative serology for HIV were included as controls. The following determinations were done in all cases: CD3+, CD4+, CD8+, CD16+CD3-, CD19+ cell counts, IL-1; IL-12, TNFalpha in serum and TNFalpha in CSF. Ten cases with cryptococcosis and AIDS were controlled three months after treatment. The average of CD4+ and NK cell counts in patients before treatment were 22/microl and 90/microl respectively; IL-1 levels were higher in the patients than in the healthy control group, conversely IL-12 levels did not show significant differences in the three studied groups. Serum concentrations of TNFalpha were higher in patients than in the control group and were not modified after treatment, conversely antifungal medication diminished IL-1 concentration and remarkably increased NK cell counts. At the same time antigen levels in serum and CSF decreased. The results obtained seem to show that the immunological alterations observed in these patients are those characteristically exhibited in severe HIV disease and that some parameters such as CD8+ cell counts lower than 200/microl, less than 50 CD4+/microl, more than 50 pg/ml of TNF-alpha and serum capsular antigen titer higher than 1:5000 seem to predict a rapidly fatal course of infection.

[Preparation and study of an exoantigen of Histoplasma capsulatum for serological reactions.]. / Preparación y estudio de un exoantigeno de la fase levaduriforme de Histoplasma capsulatum para reacciones serológicas.

The aim of this study was to determine the usefulness of a yeast-phase exo-antigen of Histoplasma capsulatum in standard serologic reactions. Three native strains of H.capsulatum which belong to Mycology Center collection were employed. They were maintained in their yeast-phase by weekly subcultures in 2% dextrose broth agar at 37 degrees C. After one week incubation yeast cells were suspended in distilled water containing thimerosal and phenylmethyl sulfonyl fluoride at a concentration of 1:5000. This suspension was left at room temperature for 72 h, then the supernatant was separated by centrifugation and it was lyophilized. Proteins and polysaccharides concentrations were determined. Immunodiffusion (ID) tests were carried out with an antigenic dilution containing 1.4 mg/ml of proteins. This exo-antigen was submitted to SDS-PAGE. Seven protein fractions were detected but only two of them showed antigenic activity against a pool of positive human sera; the molecular weights of these two proteins were 97 kDa and 66 kDa respectively. A metabolic antigen from the mycelial phase of H. capsulatum was used as control. A rabbit gammaglobulin anti-H. capsulatum was prepared and employed as positive control in serologic reactions. The antigenic capacity of ten batches of this exo-antigen was studied by ID and counterimmunoelectrophoresis (CIE) tests using serum samples of 20 hamsters experimentally infected by intracardiac inoculation of the yeast-phase of H. capsulatum. All tests presented positive results after three weeks of the infection. Fifty sera from patients suffering progressive histopasmosis were analyzed: ID, CIE and complement fixation (CF) tests were performed in all cases. HIV negative patients presented 7/7 (100%) positive reactions with the yeast-phase exoantigen and 5/7 (71.4%) with histoplasmin. In HIV positive patients CIE and CF were the most sensitive serologic tests, they gave positive results in 15/43 cases (34.8%) with the yeast-phase exo-antigen and in 7/43 cases (13.9%) with histoplasmin. Sera from 10 patients with paracoccidioidomycosis, aspergillosis and candidiasis respectively were studied by ID with the aim of detecting serologic cross reactions. No cross reaction was detected in these serum samples. This yeast-phase exo-antigen of H. capsulatum is more sensitive than and equally as specific as control histoplasmin.

[Study of some epidemiological aspects of 253 cases of cryptococcosis.]. / Estudio de algunos aspectos epidemiológicos de 253 casos de criptococosis.

Some epidemiological characteristics of 253 cases of cryptococcosis (CRY) diagnosed between 1981 and 1993 in the Muñiz Hospital (MH) of Buenos Aires City, were studied. The incidence of CRY associated with AIDS (CRY+AIDS) in the MH during 1983-1993, could be divided into 3 periods: between 1983 and 1988 1-3 cases a year were diagnosed; during 1989-91, the number of cases dopubled annually and in 1992-93 the annual increment was lower. CRY associated with predisposing causes other than AIDS (CRY+non AIDS) exhibited an annual incidence of 0-3 cases during the whole period studied. CRY was more frequent in males (86%). The difference between sexes was more evident in CRY+AIDS patients (88% males) than CRY+non AIDS ones (65% males). The median age (MA) of the studied population was 28 (range 10-71) years; 27 (10-48) in women and 29 (17-71) in men. CRY+AIDS and CRY+non AIDS patients exhibited a MA of 29 (17-51) and 40 years (10-71), respectively. AIDS was the predisposing factor in 92% of patients; 65% of them were intravenous drug abusers and 22% homosexual males, with a MA of 27 (17-40) and 33 (23-55) years, respectively. Cryptococcus neoformans var. neoformans was isolated from all CRY+AIDS and 79% of CRY+non AIDS patients and the gattii variety (Serotype B) produced 4 (21%) cases of CRY+non AIDS.

[Respiratory cryptococcosis in HIV positive patients.]. / Criptococcosis respiratoria en pacientes VIH positivos.

Although the lungs are the portal of entry of the infection, respiratory manifestations of AIDS related cryptococcosis have not been very well studied. The lack of typical findings in clinical and roentgenographic studies and the difficulties in the interpretation of the isolation of Cryptococcus neoformans from bronchial secretions, is probably the explanation for the lack of interest on this subject. The clinical and microbiological findings of 22 HIV positive patients, who presented C. neoformans in their respiratory tract clinical samples, are presented. Seventeen were males and 5 females, their age average was 30.8 years (21-50 years) and the following risk factors for HIV infection were detected: intravenous drug abuse 18, heterosexuals with several sexual partners two, one female prostitute and 1 homosexual man. All patients, except three, showed less than 100 CD4+ cells per microl. The following symptoms were observed: fever, cough, mucoid expectoration and chest ache. Roengenographic studies presented diffuse infiltrative patches in eleven cases, pulmonary cavities in three, pseudotumoral nodules in two, pneumonic infiltration in two and pleural effusion in four patients. C. neoformans was observed and/or isolated from sputum in nine patients, from bronchoalveolar lavage in seven, from lung biopsy in one and from pleural effusion in four cases. Blood cultures for C. neoformans were positive in 13 cases, urine cultures in 10 and in 11 patients C. neoformans was isolated from C.S.F. The latex agglutination tests for C. neoformans capsular polysaccharide rendered positive results in serum samples from 19 patients and from C.S.F. in 14 cases. Seven cases also presented active tuberculosis. According to these findings, it seems that the isolation of C. neoformans from bronchial secretion of HIV positive patients is a signal of disseminated cryptococcosis. It is important to isolate C. neoformans or detect its capsular antigen from other clinical samples in order to confirm the diagnosis of disseminated cryptococcosis. As observed in other studies, pleuropulmonary cryptococcosis does not present a typical clinical pattern.

[Study of eight cases of coccidioidomycosis in a hospital of Buenos Aires.]. / Estudio de ocho casos de coccidoidomicosis en un hospital de Buenos Aires.

Some clinical, epidemiological and diagnostic aspects from eight patients with chronic coccidioidomycosis (five pulmonary and three disseminated), diagnosed in the Muñiz Hospital, were retrospectively analyzed. At diagnosis, lung cavitation and hemoptysis were present in five and four patients, respectively. Smoking (three cases) and alcoholism (two cases) were the most frequent predisposing factors. Diagnosis was achieved by microscopy and cultures from sputum (five cases), tongue and lymph node biopsies and scraping of cutaneous lesion achieved diagnosis. At diagnosis, most patients had positive coccidioidin skin test and serology. Four patients were born within the endemic area and two worked in contact with the soil of the same area.

[Evaluation of a new chromogenic medium (Candida ID) for the isolation and presumptive identification of Candida albicans and other medically important yeasts]. / Evaluación micológica de un nuevo medio de cultivo cromógeno (Candida ID) para el aislamiento e identificación presuntiva de Candida albicans y otras levaduras de interés médico.

Candidiasis is a frequent human infection caused mainly by Candida albicans. However, other species are emerging as important pathogens, as Candida glabrata, Candida parapsilosis, Candida tropicalis, Candida krusei or Candida guilliermondii. Rapid identification of clinical isolates could facilitate diagnosis and treatment. Candida ID (bioMerieux, Spain) is a new medium for the isolation and presumptive identification of yeasts: C. albicans grows as blue colonies, and C. tropicalis, C. guilliermondii, Candida kefyr and Candida lusitaniae as pink ones. The utility of Candida ID was evaluated with more than 700 clinical isolates and type culture collection strains from different genera including Candida, Cryptococcus, Saccharomyces, and Rhodotorula. Presumptive identification was confirmed by germ tube test, microscopic morphology and chlamydoconidia production on corn meal agar and carbohydrate assimilation on API-ATB ID 32C or Vitek (bioMerieux). Growth on Candida ID was rapid (18-24 h) for most of the yeast strains tested. Sensitivity and specificity of identification of C. albicans was significantly high (>98%), since a very low number of isolates were found to be false negative or false positive. A better result was obtained for species growing as pink colonies (>99.5%). Detection of different species of medical important yeasts was easy with Candida ID, as perfectly distinct colors and textures of colonies were observed on this medium. Candida ID allowed the discrimination between C. glabrata (creamy and smooth) and C. krusei (rough and white) colonies. Other species showed different colony textures and colours, white being the predominant colour. Candida ID was very useful for the presumptive identification C. albicans isolates.

Disseminated histoplasmosis and AIDS: clinical aspects and diagnostic methods for early detection.

Disseminated histoplasmosis in AIDS patients is the focus of this paper. Cutaneous lesions are reported as a frequent clinical sign. Bone marrow aspiration and biopsy, blood cultures (lysis-centrifugation technique), bronchoalveolar lavage, and skin lesion scrapings are the most effective diagnostic methods. The identification of a specific antigen in blood and urine may be a rapid means of evaluation and follow-up of patients with this disease.

Flucytosine+fluconazole association in the treatment of a murine experimental model of cryptococcosis.

The efficacy of flucytosine (5-FC) and fluconazole (FLU) association in the treatment of a murine experimental model of cryptococcosis, was evaluated. Seven groups of 10 Balb C mice each, were intraperitoneally inoculated with 10(7) cells of Cryptococcus neoformans. Six groups were allocated to receive 5-FC (300 mg/kg) and FLU (16 mg/kg), either combined and individually, by daily gavage beginning 5 days after the infection, for 2 and 4 weeks. One group received distilled water and was used as control. The evaluation of treatments was based on: survival time; macroscopic examination of brain, lungs, liver and spleen at autopsy; presence of capsulated yeasts in microscopic examination of wet preparations of these organs and cultures of brain homogenate. 5-FC and FLU, individually or combined, significantly prolonged the survival time of the treated animals with respect to the control group (p < 0.01). Animals treated for 4 weeks survived significantly longer than those treated for 2 weeks (p < 0.01). No significant differences between the animals treated with 5-FC and FLU combined or separately were observed in the survival time and morphological parameters. The association of 5-FC and FLU does not seem to be more effective than 5-FC or FLU alone, in the treatment of this experimental model of cryptococcosis.

[In vitro susceptibility of Cryptococcus strains to 5 antifungal drugs]. / Susceptibilidad "in vitro" de cepas de Cryptococcus a 5 drogas antifúngicas.

A comparative study of the "in vitro" susceptibility of 24 Cryptococcus strains to 5 antifungal drugs (amphotericin B, 5 fluorocytosine, miconazole, itraconazole and ketoconazole), was carried out. These strains were grouped according to species, varieties and isolation's origins. The minimum inhibitory concentration (M.I.C.) was determined by the agar dilution technique in yeast nitrogen base agar with dextrose. The mean geometrical of the M.I.C. values of each group was compared with the others. The results obtained were homogeneous with the only exception of the "non neoformans" strains, in which, higher M.I.C. to 5 fluorocytosine values were detected.

[The epidemiological characteristics of 105 cases of cryptococcosis diagnosed in the Republic of Argentina between 1981-1990]. / Caracteristicas epidemiologicas de 105 casos de criptococosis diagnosticados en la República Argentina entre 1981-1990.

Some epidemiological characteristics of 105 cases of cryptococcosis diagnosed in Argentina, between 1981-1990, were studied. Until 1987, the annual number of cases was 4-8. It has increased since 1988 by AIDS influence and reached 35 cases in 1990. The annual number of cases non associated with AIDS has remained in 3-7 cases. The age median of the all patients, AIDS and non AIDS associated cases was 30, 30 and 45 years old, respectively. The 20-39 years old group was the most affected and the age distribution was different in AIDS and non AIDS population. Masculine predominance was more evident in AIDS than in non AIDS associated cases. The predisposing factor was AIDS in 57 patients, another different factor in 20 and unknown in 28 cases. The indirect estimation of the percentage of AIDS cases with cryptococcosis was > or = 6.19% (57 cases in 920 HIV+) during 1981-1990 this period. Cryptococcus neoformans variety neoformans was isolated from 101 cases and the gattii variety (serotype B) from only 4. The obtained data are similar to those observed in Europe and United States.

Acute disseminated histoplasmosis and endocarditis.

Acute disseminated histoplasmosis is a frequent condition in HIV carriers. Thirty-five cases of endocarditis caused by Histoplasma capsulatum have been reported in international literature, and all these descriptions correspond to a context of subacute disseminated histoplasmosis. This paper presents the case of a HIV-positive patient with fever, dyspnea, weight loss, vomiting and polyadenopathies to whom histoplasmosis was diagnosed following blood-cultures and isolation of the agent responsible for cutaneous lesions, and in whom aortic-valve vegetations were found during an echocardiogram. The patient was treated with amphotericin B and had a good outcome; subsequent echocardiograms showed no vegetations. Literature on the subject is reviewed, with special emphasis on diagnosis and treatment of previously described cases.

[E.L.I.S.A. in human coccidioidomycosis]. / E.L.I.S.A. en coccidioidomicosis humana.

An E.L.I.S.A. test for antibody detection, with an exo-antigen of Coccidioides immitis was standardized in 67 humans sera diluted in 1/1000, 1/2000, 1/4000 and 1/8000. Eighteen sera from mycologically proved cases of coccidioidomycosis were studied: 5 were negative and 13 were positive in some dilutions. 3/26 sera of healthy persons who presented positive skin tests with coccidioidin were positive and the other 23 sera did not have positive reactions. None of the 15 sera of healthy human exhibited positive E.L.I.S.A. Serum samples of 8 patients suffering other deep mycosis were studied, 4 of them presented cross-reactions in E.L.I.S.A. tests. E.L.I.S.A. test seems to be a useful serologic technique for antibody detection in anticomplementary serum samples or when a low concentration of antibodies should be detected. As it is very sensitive, cross-reactions with other mycoses are frequent, thus the use other more specific serologic technique together E.L.I.S.A. is recommended.