SPIROPLASMA-CHRYSOPICOLA SP-NOV, SPIROPLASMA-GLADIATORIS SP-NOV, SPIROPLASMA-HELICOIDES SP-NOV, AND SPIROPLASMA-TABANIDICOLA SP-NOV, FROM TABANID (DIPTERA, TABANIDAE) FLIES

Four spiroplasma strains, DE-1(T), TG-1(T), TABS-2(T), and TAUS-1(T), all of which were isolated from deerflies or horseflies (Diptera: Taba nidae), were serologically distinct from previously described spiropla sma species, groups, and subgroups. Strain DF-1(T) originated from a M aryland deerfly (Chrysops sp.); strain TG-1(T) was isolated from a Mar yland horsefly (Tabanus gladiator); strain TAUS-IT originated from a m ember of the Tabanus abdominalis-limbatinevris complex of horseflies c ollected in Maryland; and strain TABS-2(T) was isolated from a horsefl y (Tabanus abactor) collected in Oklahoma. Cells of all of the strains appeared to be helical and motile when they were examined by dark-fie ld microscopy. Cells of strain DF-1(T) growing in M1D medium were shor t helices with less than six turns; the helical cells of the other str ains were long and usually had six or more turns. The short cells of s train DF-1(T) passed through 450- and 300-nm filter pores with no redu ction in titer, but the longer cells of the other strains were partial ly retained by 450-nm-pore-size filters. Electron microscopic examinat ion of all of the strains revealed wall-less cells surrounded only by a single cytoplasmic membrane. All of the strains grew well in SP-4 li quid media and in conventional mycoplasma or M1D media supplemented wi th horse or fetal bovine serum. Strains TABS-2(T), TAUS-1(T), and DF-1 (T) required serum or sterol for growth, but strain TG-1T was able to grow in the absence of serum or sterol. The optimum temperatures for g rowth of the four strains varied from 30 to 32 degrees C, and growth o ccurred at 10 to 37 degrees C. All of the strains catabolized glucose but did not hydrolyze urea. Only strain DF-1(T) hydrolyzed arginine. T he guanine-plus-cytosine contents of the DNAs of the strains were: DF- 1(T), 29 + 1 mol%; TG-1(T), 26 +/- 1 mol%; TABS-2(T), 27 +/- 1 mol%; a nd TAUS-1(T), 26 +/- mol%. The genome sizes of strains DF-1(T) and TAU S-1(T) were 1,270 and 1,375 kbp, respectively. Strain DE-1 (= ATCC 432 09), the representative of spiroplasma subgroup VIII-2, is designated the type strain of a new species, Spiroplasma chrysopicola. We also pr opose that strain TG-1(T) (= ATCC 43525(T)), the designated representa tive of group XXIII, should be placed in a new species, Spiroplasma gl adiatoris. In addition, group XXXII spiroplasma strain TABS-2 (= ATCC 51746) is designated the type strain of Spiroplasma helicoides sp. nov ., and group XXXIII representative strain TAUS-1 (= ATCC 51747) is des ignated the type strain of another new species, Spiroplasma tabanidico la.