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Spiroplasma litorale sp. nov., from Tabanid Flies (Tabanidae: Diptera) in the Southeastern United States

MEGHNAD KONAI¹, ROBERT F. WHITCOMB^2,*, FRANK E. FRENCH³, JOSEPH G. TULLY⁴, DAVID L. ROSE⁴, PATRICIA CARLE⁵, JOSEPH M. BOVé⁵, KEVIN J. HACKETT¹, ROBERTA B. HENEGAR², TRUMAN B. CLARK^1, and DAVID L. WILLIAMSON⁶

¹ Insect Biocontrol Laboratory, Beltsville Agricultural Research Center, U.S. Department of Agriculture, Beltsville, Maryland 20705; Department of Biology, Georgia Southern University, Statesboro, Georgia 30460
² Vegetable Laboratory, Beltsville Agricultural Research Center, U.S. Department of Agriculture, Beltsville, Maryland 20705; Department of Biology, Georgia Southern University, Statesboro, Georgia 30460
³ Agricultural Research Service, Beltsville Agricultural Research Center, U.S. Department of Agriculture, Beltsville, Maryland 20705; Department of Biology, Georgia Southern University, Statesboro, Georgia 30460
⁴ Mycoplasma Section, Laboratory of Molecular Microbiology, National Institute of Allergy and Infectious Diseases, Frederick Cancer Research Facility, Frederick, Maryland 21702
⁵ Laboratoire de Biologie Cellulaire et Moléculaire. Institut Nationale de Recherche Agronomique, 33883 Villenave d'Ornon, France
⁶ Department of Anatomical Sciences, State University of New York, Stony Brook, New York 11794⁶

^* Corresponding author, Mailing address: Vegetable Laboratory, Range 2, GH 3-1, BARCW, Beltsville, MD 20705. Phone: (301) 504-8339. Fax: (301) 504-6017.

ABSTRACT

Spiroplasma strain TN-1^T (T = type strain), a strain serologically distinct from other spiroplasma species, groups, and subgroups, was isolated from the gut of a horsefly (Tabanus nigrovittatus) from a barrier island off the coast of North Carolina. Related strains were isolated from other Tabanus spp., T. atratus, T. americanus, T. gladiator, T. lineola, T. sulcifrons, and T. zythicolor, from coastal Georgia. Cells of strain TN-1^T in culture were helical and motile with an average of 5 to 10 helical turns per cell. Electron microscopic studies determined that the cells of strain TN-1^T were surrounded by a single cytoplasmic membrane, and there was no evidence of a cell wall. The spiroplasma grew well in MID and SP-4 liquid media. Serum fraction (1%) medium and conventional horse serum medium also supported growth of strain TN-1^T. Fried-egg colonies were not produced; instead, the strain produced small diffuse colonies that were surrounded by satellite growth. The optimum temperature for growth was 32°C, but multiplication was observed at temperatures from 10 to 41°C. The doubling time at the optimum temperature (32°C) was 1.6 h. No growth was observed at 5 or 43°C. Spiroplasma strain TN-1^T passed through 220-nm filter pores but failed to pass through 100-nm filter pores. Strain TN-1^T catabolized glucose but hydrolyzed neither arginine nor urea. The guanine-plus-cytosine content of the DNA was about 25 ± 1 mol%, and the genome size was 1,370 kbp. Based on results from this study and previously published data, strain TN-1^T (= ATCC 43211) (group XVIII) is designated the type strain of a new spiroplasma species, Spiroplasma litorale.