PMC5857371_01
Clinical finding of infected horse. (a) Nasal mass lesion with ulcerative wound. (b) Coral-like necrotic tissues (kunkers) from the nasal mass.
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F1
The mass was approximately 10 cm from the muzzle; the size was approximately 3 cm in diameter with an ulcerative wound and serosanguineous discharge (Fig. 1a). Two weeks after admission, many coral-like necrotic tissues were drained from the ulcerative nodule (Fig. 1b).
Diagnostic methods used for pythiosis confirmation. (a) Histopathologic demonstration of tissue biopsy: left, Gomori's methenamine silver stain showing numerous sparse septate hyphae; right, immunostaining of the biopsy tissue with specific antibody to P. insidiosum showing positive staining and a large number of branching hyphae. (b) Culture of P. insidiosum isolated from kunkers on Sabouraud dextrose agar supplemented with chloramphenicol for 15 days. (c) Detection of IgG against P. insidiosum by immunochromatographic test using P. insidiosum antigen; A, positive control; B, negative control; C, horse serum sample; C, control line; T, test line.
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F2
Immunohistological staining of the biopsy tissue with specific antibody to P. insidiosum tested positive and showed a large number of branching hyphae (Fig. 2a). Fungal cultures from fresh kunkers plated on Sabouraud dextrose agar supplemented with 0.05 mg ml-1 chloramphenicol were incubated in a 37  C aerobic condition for 15 days and presented a submerged hyaline colony (Fig. 2b). The sample showed positive results like a positive control from a human patient pythiosis serum (Fig. 2c).