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Spiroplasma montanense sp. nov., from Hybomitra Horseflies at Northern Latitudes in North America

¹Vegetable Laboratory, U.S. Department of Agriculture, Beltsville, Maryland 20705
²Department of Biology, Georgia Southern University, Statesboro, Georgia 30460
³Mycoplasma Section, Laboratory of Molecular Microbiology, National Institute of Allergy and Infectious Diseases, Frederick Cancer Research Facility, Frederick, Maryland 21701
⁴Laboratoire de Biologie Cellulaire et Moléculaire, Institut Nationale de Recherche Agronomique, Villenave d'Ornon, France
⁵Insect Biocontrol Laboratory, U.S. Department of Agriculture, Beltsville, Maryland 20705
⁶Department of Anatomical Sciences, State University of New York, Stony Brook, New York 11794

^* Corresponding author. Mailing address: Vegetable Laboratory, HH3, Range 2, BARCW, Beltsville, MD 20705. Phone: (301) 504–8339. Fax: (301) 504–6017.

ABSTRACT

Spiroplasma strain HYOS-1^Twas isolated from a tabanid fly, Hybomitra opaca. The organism was serologically distinct from other spiroplasma species, groups, and subgroups and was recently designated the representative of spiroplasma group XXXI. The cells of strain HYOS-1^T, as determined by light microscopy, were long motile helices. Electron microscopic examination revealed wall-less cells delimited by a single membrane. The cells passed through 450- and 300-nm filter pores with a 10-fold reduction in titer, but failed to pass through 100-nm pores. Strain HYOS-1^Tgrew very well in most conventional medium formulations for spiro-plasmas or other mollicutes. The organism grew at temperatures ranging from 5 to 41°C, and the optimum temperature was 32°C. The doubling time at the optimum temperature was 0.7 h, one of the shortest values obtained for members of the genus Spiroplasma. The strain catabolized glucose and hydrolyzed arginine but not urea. Growth of the organism was stimulated by cholesterol and serum, but the strain was nevertheless able to grow in the absence of sterols or serum. The guanine-plus-cytosine content of the DNA was about 28 ± 1 mol%, and the genome size was 1,225 kbp. On the basis of the experimental results reported here and previously reported data, group XXXI strain HYOS-1 (= ATCC 51745) is designated the type strain of a new species, Spiroplasma montanense.