Spiroplasma taiwanense Abalain-Colloc et al.
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1. Follow instructions as suggested for the culturing of Mollicutes:
PROCEDURES FOR PROPAGATING MOLLICUTES:
a) Open the vial according to the enclosed instructions.
b) Using a Pasteur or 1.0 ml pipette, withdraw approximately 0.5 to 1.0 ml from a tube containing 5.0 ml. Rehydrate the entire pellet.
c) Aseptically transfer this aliquot back into the tube. Mix well.
d) Make serial dilutions by transferring 0.5 ml from the original tube to a tube containing 4.5 ml. Repeat process by transferring 0.5 ml from the second to a third tube, etc. Dilutions are important, not only for titration purposes, but also to keep culture in varying stages of growth. Many strains will die out rapidly once acid or alkaline conditions are reached. It is recommended to prepare several dilutions from the initial tube as the cryoprotectant used in the freeze‑drying process often inhibits growth.
e) Use an uninoculated tube of broth to serve as a control.
f) Plates may be inoculated to check colonial morphology. You can also spot each dilution on the surface of plate (4 or more/plate) to determine the number of colony-forming units. However, not all strains do well on solid medium.
g) Incubate all tubes and plates under the recommended conditions and appropriate temperature. The time necessary for growth will vary from strain to strain. Growth on plates generally requires additional incubation.
h) Depending on the medium used, growth will be indicated by increased turbidity, a color change, or
2. Tubes and plates are incubated aerobically. The incubation temperature is 30oC.
3. Growth occurs in 24 to 48 hours. Additional incubation is required for colonies to appear on solid medium. The broth will turn from red-orange to yellow.
Store vials at freezer temperatures until ready to use.
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Tully JG, et al. Revised group classification of the genus Spiroplasma (Class Mollicutes), with proposed new groups XII and XXIII. Int. J. Syst. Bacteriol. 37: 357-364, 1987.
Abalain-Colloc ML, et al. Spiroplasma taiwanense sp. nov. from Culex tritaeniorhynchus mosquitoes collected in Taiwan. Int. J. Syst. Bacteriol. 38: 103-107, 1988.