Epizootic cutaneous pythiosis in beef calves

Enzootic bovine granulomatosis (EBG) is a disease that affects beef calves in the flooded Savanna regions of Venezuela. Although Pythium insidiosum was originally the suspected etiologic agent, nothing was done to demonstrate its presence in the infected animals. The objective of this study was to investigate the etiologic agent causing cutaneous lesions in a group of 63 calves diagnosed with EBG. The collected samples were analyzed by histopathology, immunohistochemistry, serology, and mycological techniques. The clinical manifestations of BEG included ulcerated and swollen granulomatous lesions on the limbs of young calves. Histopathological studies revealed an eosinophilic inflammatory reaction with neutrophils, giant cells and branching hyphae with the Splendore-Hoeppli phenomenon. Immunofluorescence analysis identified the branching hyphae as elements of the fungal-like organism P. insidiosum. Wet mounts in 10% KOH revealed hyaline slender branching hyaline coenocytic hyphae, but cultures from those samples were negative. Anti-P. insidiosum immunoglobulins were also demonstrated in the collected sera of the affected bovines using latex agglutination, immunodiffusion, ELISA, and Western Blot. The histopathological, immunohistochemical, and serological tests showed for the first time that P. insidiosum could be involved in epizootic bovine pythiosis.