PLATE XXIXIN a continuing study on the habitat of the agents of deep mycoses, suspensions of soil and plant debris are injected into guinea-pigs and mice. Several moulds and actinomycetes have been observed in, or cultivated from, lesions produced in the experimental animal or their apparently normal tissues. During this work we observed that Pseudomonas aeruginosa (Ps. pyocyanea) may produce actinomycetoid granules and the histopathological picture of botryomycosis.
MATERIALS AND METHODSOur first observations were accidental. If our purpose had been to isolate Ps. aeruginosa we should not have used the method described below, which is a modification of that of Emmons (1949). Seventy-four samples of soil and 51 of plant debris were taken with sterile wooden tongue-depressors and placed in plastic bags. The samples of plant debris included 24 samples of powdery or fibrous debris from rotten palm-tree trunks, 8 nests of armadillos and wild rodents, built up with grass blades, and 19 samples taken from hollows in tree trunks.A 5-ml volume of each material was placed in a sterile flask with 25 ml isotonic saline solution and glass beads and shaken vigorously for 5 min. The suspension was then left for 5-10 min.; heavy particles settled and the plant debris floated. An 8-ml volume of the intervening small-particle suspension was pipetted into a test-tube with 2 ml of a solution containing 5 mg penicillin and 4 mg dihydrostreptomycin per ml. Guinea-pigs received 0.2 ml of this suspension in each testicle, and male mice were given 0-25-ml doses intraperit oneally.The exudates from the lesions produced in these animals were examined in stained smears and also in wet preparations directly between slide and coverslip. Sections from tissues fixed in 4 per cent. formaldehyde and embedded in paraffin were stained with haematoxylin and eosin.The following results refer to observations and experiments with 3 strains of Ps. aeruginosa; 2 were isolated from plant debris and 1 strain was isolated from man.
RESULTS
Experiments with the first strain isolated from plant debrisThe inoculated material was taken from the decayed trunk of a fallen palm-tree (Bufia yatay) of which the inner portion was soft, spongy and at some areas hollow. Skunks, armadillos and small wild rodents had penetrated and some had made their nests with grass. Larvae of beetles, ants and other arthropods were found.After intratesticular injection of the sample, the test guinea-pig showed bilateral orchitis; the animal was killed 20 days after the inoculation. Filaments of a semi-acid-fast actinomycete were seen in the exudate from the left testicle; granules were not found, but cultures yielded Nocardia brasiliensis. Actinomycetoid granules with clubs were seen in the pus of the right testicle when examined between slide and coverslip; filaments of actinomycetes were not seen; a Gram-negative bacillus was seen in smears, and culture produced a growth of -
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