SPIROPLASMA-LAMPYRIDICOLA SP-NOV, FROM THE FIREFLY-BEETLE PHOTURIS-PENNSYLVANICUS

Spiroplasma strain PUP-1(T) was isolated from the gut fluids of a fire fly beetle (Photuris pennsylvanicus) collected in Maryland, Cells of t he strain were shown by dark-field microscopy to be helical, motile fi laments. Ultrastructural examination by electron microscopy revealed f ilamentous cells bounded by a single cytoplasmic membrane and no evide nce of a cell wall, The cells were not sensitive to 500 U of penicilli n per mi and grew under aerobic conditions in M1D, SP-4, and M-2 broth formulations, as well as in conventional mycoplasma medium, The doubl ing times at 15, 20, 25, and 30 degrees C were 83.1, 32.0, 14.9, and 9 .8 h, respectively. Suboptimal growth occurred at 37 degrees C, and no growth was apparent in cultures maintained at 10 or 40 degrees C, The organism required cholesterol for growth and produced acid from gluco se, fructose, and trehalose; arginine and urea were not hydrolyzed, Th e results of previous serological analyses of strain PUP-1(T) indicate d that the organism was not related to the then currently established Spiroplasma species or group representatives, and the organism was cla ssified as the representative of group XIX, Additional testing of stra in PUP-1(T) against recently recognized Spiroplasma species or group r epresentatives by both metabolism inhibition and deformation tests con firmed the unique serological status of the organism, The guanine-plus -cytosine content of the DNA was 26 +/- 1 mol%, and the genome size wa s 1,375 kbp. These values clearly differentiate strain PUP-1(T) from g roup XXI strain W115, with which it cross-reacted reciprocally at a lo w level in deformation and metabolism inhibition tests, We propose tha t strain PUP-1 (= ATCC 43206) should be recognized as the type strain of a new species, Spiroplasma lampyridicola.