Spiroplasma lampyridicola sp. nov., from the firefly beetle Photuris pennsylvanicus

1 Department of Agricultural Sciences and Department of Microbiology, School of Veterinary Medicine, Tuskegee University, Tuskegee, Alabama 36088 2 Mycoplasma Section, Laboratory of Molecular Microbiology, National Institute of Allergy and Infectious Diseases, Frederick Cancer Research Facility, Frederick, Maryland 21702 3 Insect Biocontrol Laboratory, Vegetable Laboratory, USDA Agricultural Research Service, BARCW, Beltsville, Maryland 20705 7 Vegetable Laboratory, USDA Agricultural Research Service, BARCW, Beltsville, Maryland 20705 4 Department of Anatomical Sciences, State University of New York, Stony Brook, New York 11794 5 Laboratoire de Biologie Cellulaire et Moléculaire, Institut Nationale de Recherche Agronomique, 33883 Villenave d'Ornon Cedex, France 6 Department of Biology, Georgia Southern University, Statesboro, Georgia 30460 * Corresponding author. Mailing address: Vegetable Laboratory, GH3-1, Bldg. 010, Range 2, BARCW, Beltsville, MD 20705. Phone: (301) 504-8339. Fax: (301) 504-6017. ABSTRACT Spiroplasma strain PUP-1 T was isolated from the gut fluids of a firefly beetle ( Photuris pennsylvanicus ) collected in Maryland. Cells of the strain were shown by dark-field microscopy to be helical, motile filaments. Ultrastructural examination by electron microscopy revealed filamentous cells bounded by a single cytoplasmic membrane and no evidence of a cell wall. The cells were not sensitive to 500 U of penicillin per ml and grew under aerobic conditions in M1D, SP-4, and M-2 broth formulations, as well as in conventional mycoplasma medium. The doubling times at 15, 20, 25, and 30°C were 83.1, 32.0, 14.9, and 9.8 h, respectively. Suboptimal growth occurred at 37°C, and no growth was apparent in cultures maintained at 10 or 40°C. The organism required cholesterol for growth and produced acid from glucose, fructose, and trehalose; arginine and urea were not hydrolyzed. The results of previous serological analyses of strain PUP-1 T indicated that the organism was not related to the then currently established Spiroplasma species or group representatives, and the organism was classified as the representative of group XIX. Additional testing of strain PUP-1 T against recently recognized Spiroplasma species or group representatives by both metabolism inhibition and deformation tests confirmed the unique serological status of the organism. The guanine-plus-cytosine content of the DNA was 26 ± 1 mol%, and the genome size was 1,375 kbp. These values clearly