Experimental Pythium aphanidermatum Infection in Rabbits.

Pythium aphanidermatum is a common plant pathogen, reportedly capable of human infection. This study investigated whether P. aphanidermatum can infect domestic rabbits. Four rabbits were inoculated with P. aphanidermatum subcutaneously and evaluated for 30 days. Blood and tissue samples were collected from all animals. The animals developed palpable nodules 5-10 days after inoculation. Necropsy revealed caseous nodular lesions characterized histopathologically by pyogranulomatous inflammation containing hyphae in the necrotic areas. P. aphanidermatum was isolated from the caseous material and anti-P. aphanidermatum antibodies were detected by serological examination. This is the first report of experimental P. aphanidermatum infection in mammals. The results demonstrate that this oomycete can produce infection at the inoculation site and induce an immune response. The results suggest that P. aphanidermatum may be undergoing adaptation to vertebrate hosts.

Protein profile of Brazilian Pythium insidiosum isolates.

Pythium insidiosum is an important aquatic Oomycota that causes pythiosis in mammals, especially horses, dogs, and humans; these inhabit marshy environments in tropical and subtropical areas. The aim of this study was to determine the protein profile, as well as identify likely immunodominant proteins, of Brazilian P. insidiosum isolates from southern Brazil, an important equine pythiosis endemic area. P. insidiosum isolates (horses, n = 20 and dogs, n = 02) were analyzed by SDS-PAGE and Western blot techniques. Horse, cattle, dog, and rabbit sera of both diseased and healthy animals were used to identify P. insidiosum proteins. SDS-PAGE protein profile detected antigens of molecular weights ranging from 100 to 20 KDa. Dog isolates revealed a protein profile similar to that of horse isolates. Anti-P. insidiosum antibodies in the sera of the four species could recognize proteins of different molecular weights (∼74 KDa to ∼24 KDa), and proteins ∼50-55 KDa and ∼34 KDa were shown to be immunodominant. Furthermore, ∼74 KDa, ∼60 KDa, ∼30 KDa and ∼24 KDa proteins were poorly recognized by host species antibodies. The Brazilian P. insidiosum isolates analyzed showed a similar protein profile; however, further studies are essential for the identification and characterization of proteins expressed by P. insidiosum, and an evaluation of the immunological profile of hosts susceptible to this Oomycota is necessary.

In Vitro Susceptibility of Pythium insidiosum to Melaleuca alternifolia, Mentha piperita and Origanum vulgare Essential Oils Combinations.

Pythium insidiosum is the etiologic agent of pythiosis, a severe and emerging disease that affects mammals. Failure of conventional antifungal therapies is partially justified by the absence of ergosterol in the plasma membrane of this oomycete. Despite research advancement, the treatment of pythiosis has not been not fully established. The present study investigated the in vitro susceptibility profile of Brazilian isolates of P. insidiosum (n = 20) against Melaleuca alternifolia, Mentha piperita and Origanum vulgare essential oils, and their combinations. Susceptibility tests were performed according to CLSI M38-A2 protocol, and combinations were evaluated by the microdilution cherkerboard method. All tested essential oils showed antimicrobial activity against P. insidiosum, and the greatest activity of O. vulgare was highlighted. Synergistic and/or indifferent effect was observed for all combinations evaluated, especially the M. piperita and O. vulgare combination, which showed 65 % synergism. This is the first study to report in vitro combinations of essential oils against P. insidiosum indicating the susceptibility of this oomycete to M. alternifolia, M. piperita and O. vulgare essential oils, as well as their combinations.