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Histologic and molecular identification of disseminated Histoplasma capsulatum in a captive brown bear (Ursus arctos)

Highland, Margaret A., Chaturvedi, Sudha, Perez, Michael, Steinberg, Howard, Wallace, Roberta
ribosomal RNA, diaphragm, Candida, genes, necropsy, hibernation, thoracic cavity, antibodies, DNA, internal transcribed spacers, histoplasmosis, lymph nodes, Ursus arctos, radiography, yeasts, Histoplasma capsulatum, adrenal glands, Aspergillus, histopathology, Blastomyces, quantitative polymerase chain reaction, anorexia, inflammation, cough, gene targeting, Coccidioides, bacteria
A 33-year-old brown bear (Ursus arctos) was evaluated for chronic cough, partial anorexia, and lethargy in early fall of 2009. Radiographs revealed a generalized increase in interstitial density with focal lung field consolidation and air bronchograms more prevalent in the cranial lung lobes. Tracheal sputum and wash fluid grew mixed bacteria and 2 species of Candida on bacterial and fungal cultures, respectively. Serum was negative for antibodies to Aspergillus, Blastomyces, Coccidioides, and Histoplasma by semiquantitative radial immunodiffusion. Antimicrobial and antifungal treatment was administered. The bear died 1 month after entering hibernation. Gross necropsy revealed coalescent nodules and sheets of firm tan tissue covering pleural surfaces of the thoracic cavity and within pulmonary parenchyma, enlarged mesenteric lymph nodes, and intestinal ulcerations. Histopathology revealed granulomatous inflammation with intrahistiocytic yeast, consistent with Histoplasma organisms, in lung, diaphragm, mesenteric lymph nodes, intestine, and adrenal glands. Molecular analysis performed on DNA isolated from lung tissue, including conventional polymerase chain reaction (PCR) targeting the internal transcribed spacer region for the ribosomal RNA gene complex and real-time PCR targeting the gene encoding a unique region of M specific protein, identified the organism to be 100% identical to Histoplasma capsulatum with an average of 4.9 × 10⁷ gene copies per gram of tissue. The present report describes histologic and molecular techniques for diagnosing histoplasmosis.