A domestic ferret (Mustela putorius furo) was presented with lymphadenopathy and acute bilateral blindness. Cytologic evaluation and biopsy of an affected lymph node revealed pyogranulomatous lymphadenitis with intralesional yeast consistent with Cryptococcus sp. Subsequent studies demonstrated Cryptococcus gattii serotype B VGI/AFLP4 as the causative agent. The ferret was treated with fluconazole and prednisone. After one month of therapy, an improvement of the clinical symptoms was detected although blindness persisted. Seven months after presentation, the disease progressed to a severe neurologic condition, and it was euthanized. Postmortem exam revealed disseminated cryptococcosis with prominent neurologic involvement. Nasal swabs of other ferrets and humans from the same household revealed that two ferrets and two humans to be asymptomatic carriers of the same strain of cryptococcus as the necropsied ferret. These findings stress the importance of veterinary diagnostic work with pets and epidemiological investigations for disease prevention in them and in their owners.
Fusarium proliferatum caused endophthalmitis after cataract surgery. Diagnosis was established by classical microbiology and molecular biology methods (PCR and DNA typing). The treatment with local amphotericin B, oral ketoconazole, and topical natamycin was successful. CASE REPORTA 66-year-old man underwent cataract extraction and intraocular lens implantation in his right eye. Four months after surgery, he complained of eye discomfort and was prescribed a topical steroid (dexamethasone) and tobramicin. Fifteen days later, his symptoms persisted, and the eye began to show severe palpebral edema. Slit lamp examination showed conjunctival hyperemia, hypopyon, capsular fibrosis, and corneal edema, and the intraocular lens had moved to the nasal site. Intraocular pressure was 28 mm Hg, and fundus examination showed vitreous haze. An aqueous humor sample was taken using a 30-gauge needle following examination of the eye. Diagnostic techniques included standard microbiological tests (culture and stains) and PCR. The aqueous humor sample was cultured on several media including Columbia agar plates supplemented with 5% sheep blood chocolate agar, MacConkey agar, and thioglycolate broth and brain heart infusion broth (all media were from Biomérieux, biomérieux Sa, Marcy L'Etoile, France) at 37°C in ambient air. The sample was also inoculated into Sabouraud dextrose agar with chloramphenicol and incubated at 30°C. Two different PCRs were carried out: the first focused on bacterial 16S rRNA gene (16) amplification, and the second focused on specific detection of the fungal internal transcribed spacer (ITS)/5.8S DNA region (5). All the tests on the aqueous humor sample (stains, cultures, and PCRs) were negative. Next, a vitreous sample was taken, and microbiological and molecular tests were repeated. Direct smear of the vitreous sample showed septate hyphae of a filamentous fungus with neutrophils ( Fig. 1). PCR with specific fungal primers was positive, and PCR with bacterial primers was negative. This preliminary result was obtained 6 h after the sample was taken. Antifungal treatment was given as soon as the fungal hyphae were seen in the vitreous sample by direct smear.Intravitreal amphotericin B (5 g/0.1 ml) and oral fluconazole (200 mg/day) were administered the same day the sample was taken.Amplified DNA from fungal PCR was submitted for sequence analysis (PE Applied Biosystems, Foster City, Calif.) and compared to DNA sequences in the BLAST alignment program of the GenBank database (National Institutes of Health) and the EMBL fungal DNA database using Fasta3 sequence similarity searches, which allowed the species identification 48 h after the sample was obtained. DNA database comparison of the sequence showed 100% identity with Fusarium proliferatum (NRRL 31071; USDA Agricultural Research Service Collection, Peoria, Ill.), 99.8% identity with Fusarium fujikuroi, and 99.8 to 99.6% identity with Fusarium annulatum, all of which belong to the Gibberella fujikuroi complex (10, 26).When the molecular identification show...
We report the first case of human cryptococcosis due to Cryptococcus neoformans var. gattii described in our country, which was presented as brain cryptococcoma in an immunocompetent patient. An extensive sampling of the patient's environment was carried out to find the source of infection. CASE REPORTA 60-year-old heterosexual Spanish farmer came to the Hospital General de Alicante in July 2003, having suffered for several days from cephalalgia and somnolence. He had never traveled abroad. A diabetes mellitus type 2 identified 2 years previously was the only clinical antecedent of interest. Human immunodeficiency virus serology was investigated with repeated negative results. General and neurological exploration included computerized tomography scanning, which disclosed a brain mass lesion in basal ganglions. Capsulated yeasts were seen in a stereotaxic brain puncture sample, and Cryptococcus neoformans was suspected to be the causative agent. C. neoformans capsular antigen was detected in blood and cerebrospinal fluid (CSF) several times during the process (maximum values detected, 1/256 and 1/32, respectively). The yeast was cultured from a surgical drainage sample of the brain abscess. Species identification was carried out on the basis of microscopic morphology, growth at 37°C, a urease test, phenoloxydase production, and the carbohydrate assimilation pattern (Auxacolor; Bio-Rad). Further testing such as canavanina glycine bromothimol blue agar growth, serotype determination (Cryptocheck test; Iatron), and genotype analysis revealed that the strain was C. neoformans var. gatti serotype B. The strain identification and serotype were confirmed in another mycology laboratory (IMIM, Barcelona, Spain). Two antifungal drug sensitivity tests were performed (Sensititre and Etest). Both tests showed low amphotericin B (AMB) and ketoconazole MICs but different results with fluconazole (MIC sensititre , 8 g/ml; MIC Etest , 64 g/ml) and 5-flucytosine (MIC sensititre , 2 g/ml; MIC Etest , 32 g/ml). Voriconazole was only tested with the Sensititre test (MIC sensititre , 12 g/ml). Genotype analysis consisted of the study of five molecular DNA targets: internal transcribed spacer-5.8S rRNA gene sequence analysis (36), 5S rRNA gene and URA5 gene restriction fragment length polymorphism (RFLP) analysis (24), and amplification patterns of two highly repeated minisatellite sequences (M13 and GACA 4 ) (14, 24). Nucleotide sequence analysis of the internal transcribed spacer-5.8S rRNA gene confirmed the identification at a species level by comparing it to ribosomal sequence databases (EMBL and GenBank). RFLP of the URA5 gene showed a VGI molecular type (Fig. 1) which is characteristic of C. neoformans var. gattii. RFLP 5S rRNA gene and minisatellite amplification of M13 and GACA 4 also displayed molecular patterns attributable to C. neoformans var. gattii strains (14, 24).Some different antifungal therapies were followed, depending on the clinical evolution and the antigen levels (serum and CSF). Intravenous AMB at 200 mg/day (A...
We propose that Latin American immigrant patients admitted to hospital in Spain be screened for strongyloidiasis, Chagas disease and syphilis by serology.
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