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Conventional AMB was started day but discontinued shortly after development of chemical meningitis on day Acute phase reactants were normal. An Omaya reservoir was inserted day after two more times replacing the catheter. However it had to be discontinued because of recurrent anatomical and technical problems. On daya culture of a CSF sample was negative. As the patient gradually improved he was discharged on day with oral VRZ, methylprednisolone and anti-epileptics.
He was followed-up for about two years until his death. Brain MRIs on days 83 and and Materials and methodsIsolation and phenotypic identification A brain biopsy, two subsequent aspiration materials from cerebral lesions and a cerebrospinal fluid CSF sample were submitted to Deep Mycoses Laboratory on days, andrespectively.
Direct microscopic examination was carried out using Gram, EZN and multiple Giemsa stains of the imprinted tissue specimen, abscess materials and CSF sample.
All plates were sealed with parafilm to maintain adequate humidity for prolonged incubation time.
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Microscopic morphology of the isolate was examined by staining with lactophenol cotton blue and using unstained wet mounts with saline.
Periodic acid schiff PAS and hematoxylin-eosin H-E stained histopathological preparations from biopsy material of day were also examined. Observations and photomicrographs were made using an Olympus CD2 photomicroscope.Yahoo Respuestas
Phenotypical identification was made using macroscopic and microscopic characteristics of the isolate. Molecular identification The isolate was identified by sequencing of the internal transcribed spacer ITS domain of rDNA and comparison with reference strains deposited in GenBank and a research database on black fungi housed at CBS.
Tests were carried out according to the manufacturer's instructions and were repeated twice at two-day intervals. ResultsMycology Direct microscopical examination of Giemsa stained imprinted tissue slides of the biopsy and two subsequent blackish abscess materials revealed the presence of smooth-walled, ellipsoidal conidia, scarcely septated hyphae and conidial chains arising directly from the hyphae Fig.
Gram and EZN stains did not reveal any organism. In H-E and PAS stained histopathologic preparations the organism appeared in tissue as a few pieces of brown hyphae and polymorphic fungal cells. Scarcely separated hyphae a—c and intact conidial chains d with conidiogenous scars seen in Giemsa-stained abscess and imprinted tissue slides. Biopsied tissue and the first aspiration material failed to grow on all media after three weeks of incubation.
Combined histopathology and microscopy of the isolate suggested a case of cerebral phaeohyphomycosis. Microscopic examination of unstained saline and lactophenol cotton blue stained slides revealed long, occasionally branched chains of olivaceous-brown, ellipsoidal conidia.
Fungal structures seen in histopathological sections and in abscess smears were compatible with those of the isolated strain. The isolate was identified molecularly as C. The same MIC values were obtained in duplicate tests. Laboratory health risk measures All patient cultures were immediately killed by autoclaving together with biological indicators and discarded as biohazardous waste.
Environmental controls were carried out periodically in both the mycology laboratory and inside the incubators. Discussion Cladophialophora bantiana is by far the most prevalent agent of fungal brain infection in humans. The species occurs worldwide, with highest frequency on the Indian subcontinent. For this reason C. Given this pathology, understanding of the environmental habitat and route of infection are significant for both health laboratories and clinicians to assess the history and laboratory data of occasional cases, and also essential for prophylaxis.
However, only very few and paradoxal data are available and address the need of new environmental investigations. Almost all isolates known to date have been derived from human patients, some caused infections in cats and dogs, 13 once in a horse CBS 2almost always with cerebral etiology.
Verified environmental strains originated from sawdust CBS Our case concerned probable infection by traumatic inoculation from a rock in a mine shaft. The patient had a head injury with laceration of the skin while working in a coalmine, suggesting carbon dust as a possible source of infection. Several isolates had dust as a common factor, but the waterfilter location denies a xerophilic character. Oligotrophism was shared by all environmental habitats, while some had elevated temperatures.
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