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Spiroplasma clarkii sp. nov. from the Green June Beetle (Coleoptera: Scarabaeidae)

¹Insect Biocontrol Laboratory, Beltsville Agricultural Research Center, Agricultural Research Service, U.S. Department of Agriculture, Beltsville, Maryland 20705
²Laboratoire de Biologie Cellulaire et Moléculaire, Institut Nationale de Recherche Agronomique, 33883 Villenave d'Ornon Cedex, France
³Mycoplasma Section, Laboratory of Molecular Microbiology, Frederic Cancer Research Facility, National Institute of Allergy and Infectious Diseases, Frederick, Maryland 21702
⁴Department of Anatomical Sciences, State University of New York, Stony Brook, New York 11794

^* Corresponding author.

ABSTRACT

Spiroplasma strain CN-5^T (T = type strain), isolated from the gut of the cetoniine scarabaeid beetle Cotinus nitida, was serologically distinct from other spiroplasma species, groups, and subgroups. Cells of strain CN-5^T were shown by light microscopy to be helical, motile filaments. Cells in early passages exhibited strong translational motility that tended to be lost in later passages. Electron microscopy showed that the cells were bounded by a single cytoplasmic membrane with no evidence of a cell wall. The organism was not susceptible to penicillin. Strain CN-5^T grew well in SM-1, M1D, and SP-4 liquid media and on solid SP-4 medium under aerobic or anaerobic conditions. The doubling time at 30°C, the optimum temperature, was 4.3 h. The strain also grew in 1% serum fraction medium. Strain CN-5^T produced acid from glucose and catabolized arginine, but did not hydrolyze urea. The guanine-plus-cytosine content of the DNA was 29 ± 1 mol%. The genome size was 1,770 kb (1,186 MDa). Other uncloned isolates obtained from C. nitida or the cetoniine hermit flower beetle Osmoderma eremicola exhibited similar or identical serological patterns. Since no other hosts were discovered in extensive studies, strain CN-5^T (previously designated group IX) appears to represent a cluster of relatively host-specific cetoniine beetle-associated strains. Strain CN-5 (= ATCC 33827) is designated the type strain of a new species, Spiroplasma clarkii.