VIEW ARTICLE

Techniques

In Vivo Cultivation of Spiroplasmas in Larvae of the Greater Wax Moth. R. E. McCoy, University of Florida Agricultural Research Center, Fort Lauderdale 33314; M. J. Davis(2), and R. V. Dowell(3). (2)University of California, Department of Plant Pathology, Berkeley 94526, Present address: Department of Plant Pathology, Rutgers University, New Brunswick, NY 08903; (3)University of Florida Agricultural Research Center, Fort Lauderdale 33314. Phytopathology 71:408-411. Accepted for publication 11 September 1980. Copyright 1981 The American Phytopathological Society. DOI: 10.1094/Phyto-71-408.

Larvae of the greater wax moth, Galleria mellonella, supported extensive multiplication of a number of spiroplasma strains after injection with 1-μ1 doses of spiroplasma broth cultures. Spiroplasma multiplication in the insects was monitored by light-microscopic examination of hemolymph samples. Multiplication rates of the spiroplasmas in larval hemolymph varied with the spiroplasma strain. Spiroplasma citri strain G-1 and strain SR3 spiroplasmas reached titers of ca 10⁸ helices per milliliter hemolymph after 5–10 days at 30 C but were not detectable in hemolymph for at least 48 hr after injection. However, the flower isolates 23-6 and PPS1 multiplied rapidly with titers approaching 10⁷/ml of hemolymph 12 hr after injection and reaching 10⁹/ml by 24 hr. Populations remained at these levels for the duration of the larval stage and spiroplasmas were present in adult moths after pupation. Spiroplasma cells observed in hemolymph were short, actively motile helices without blebs or other degenerative signs. We feel that larvae of G. mellonella have potential value as laboratory hosts in further studies of the Mollicutes.