Abstract
The ability of several continuous tick cell culture lines to support growth of tickborne spiroplasmas (helical, wall-less prokaryotes in the classMollicutes) was assessed. Seven triturates, prepared from pools ofIxodes pacificus ticks naturally infected with theSpiroplasma sp. (group VI) organism, were retrieved from frozen (−70°C) storage and passaged in three distinct tick cell lines, in antibiotic-free tick cell culture medium alone, or in spiroplasma culture medium (SP-4 formulation). Six spiroplasma strains were recovered in the RML-19 cell line fromDermacentor variabilis, and five isolations were made in another cell line (RML-15) from this tick species. None was recovered in aRhipicephalus sanguineus cell line (RML-23), in tick cell culture medium, or in SP-4 broth medium. One of the spiroplasma isolates (Y43) was maintained through four consecutive weekly refeedings of theD. variabilis cell line and for three feedings ofR. sanguineus cells, where numbers of spiroplasmas in cell supernatants reached levels comparable to those obtained in the SP-4 medium.
A laboratory-adapted strain (SMCA) ofSpiroplasma mirum, a second helical mollicute of tick origin (the suckling mouse cataract agent), grew in three tick cell lines (RML-15, RML-23, and RML-16 cells fromD. parumapertus), in three mosquito cell lines (fromAedes albopictus, Ae. aegypti, andCulex quinquefasciatus), and in both cell culture medium alone and in SP-4 medium. The organisms survived for 1–2 weeks, but failed to multiply, in cell lines fromC. tritaeniorhynchus, Antheraea eucalypti, orXenopus laevis. Some evidence of cytopathic effect ofS. mirum on tick cell lines was seen, although growth of the organism in mosquito cell cultures was not associated with cell toxicity. The use of arthropod cell lines appears to have value in the primary isolation of arthropod- or insect-derived mollicutes and for the study of cytopathogenicity of these wall-less prokaryotes.
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Literature Cited
Bhat UKM, Yunker CE (1979) Susceptibility of a tick cell line (Dermacentor parumapertus Neumann) to infection with arboviruses. In: Kurstak E (ed) Arctic and tropical arboviruses. New York: Academic Press, pp 263–275
Bové JM (1984) Wall-less prokaryotes of plants. Annu Rev Microbiol 22:361–396
Chastel C, Gilot B, Le Goff F, Gruffaz R, Abalain-Colloc M-L (1985) Isolement de spiroplasmes en France (Savoie, Alpes du Nord) a partir de moustiques du genreAedes. CR Acad Sci [D] 300:261–266
Clark TB (1982) Spiroplasmas: diversity of arthropod reservoirs and host-parasite relationships. Science 217:57–59
Droleskey RE, Holman PJ, Craig TM, Wagner GG, Mollenhauer HH (1983) Ultrastructure ofBabesia bovis sexual stages as observed inBoophilus microplus cell cultures. Res Vet Sci 34:249–251
Garnier M, Steiner T, Martin G, Bové JM (1984). Oxido-reduction sites and relationships of spiroplasmas with insect cells in culture. Isr J Med Sci 20:840–842
Grace TDC (1962) Establishment of four strains of cells from insect tissues grown in vitro. Nature 195:788–789
Hackett KJ, Lynn DE (1985) Cell-assisted growth of a fastidious spiroplasma. Science 230:825–827
Hackett KJ, Lynn DE, Williamson DL, Ginsberg AS, Whitcomb RF (1986) Cultivation of theDrosophila sex-ratio spiroplasma. Science 232:1253–1255
Hsu SH (1971) Growth of arboviruses in arthropod cell cultures: comparative studies. I. Preliminary observations on growth of arboviruses in a newly established line of mosquito cell (Culex quinquefasciatus Say). Curr Topic Microbiol 55:140–148
Igarashi A (1978) Isolation of a Singh'sAedes albopictus cell clone sensitive to dengue and chikungunya viruses. J Gen Virol 40:531–544
Liao CH, Chen TA (1979) Cultivation of suckling mouse cataract spiroplasma in a simple medium [abstr]. Proc Am Soc Microbiol (79th annual meeting), p 82
Louis C, Quiot JM, Giannotti J, Vago C (1978) Infection expérimentale d'une lignée cellulaire d'invertébré par le procaryote intravacuolaire de type mollicute, agent de la “léthargie des coléoptères”. Ann Microbiol (Inst Pasteur) 129B:621–633
McGarrity GJ, Steiner T, Vanaman V (1983) Detection of mycoplasmal infection of cell cultures by DNA fluorochrome staining. In: Tully JG, Razin S (eds) Methods in mycoplasmology, vol 2. New York: Academic Press, pp 183–190
Peleg J (1969) Inapparent persistent virus infection in continuously grownAedes aegypti mosquito cells. J Gen Virol 5:463–471
Pudney M, Varma MGR, Leake CJ (1973) Establishment of a cell line (XTC-2) from the South African clawed toad,Xenopus laevis. Experientia 29:466–467
Pudney M, Varma MGR, Leake CJ (1978) The growth of some arboviruses in tick cell lines. In: Wilde JKH (ed) Tickborne diseases and their vectors. Edinburgh UK: University of Edinburgh, pp 490–496
Schneider I (1973) Establishment of cell lines fromCulex tritaeniorhynchus andCulex salinarius (Diptera: Culicidae). In: Proceedings of the third international colloqium on invertebrate tissue culture. Bratislava: Slovak Academy of Science, pp 121–134
Shatkin AA, Beskina SR, Medvedeva LI, Ghrohkovskaya IM (1977) Cultivation of the causative agent of enzootic ovine abortion in continuous embryonal line cells of ticks of the genusHyalomma. Med Parazitol (Moscow) 4:420–423
Singh KRP (1967) Cell cultures derived from larvae ofAedes albopictus (Skuse) andAedes aegypti (L.). Curr Sci 36:506–508
Steiner T, McGarrity GJ (1983) Mycoplasma infection of insect cell cultures. In Vitro 19:672–682
Steiner T, McGarrity GJ, Bové JM, Phillips DM, Garnier M (1983) Insect cell cultures in the study of attachment and pathogenicity of spiroplasmas and mycoplasmas. Ann Microbiol (Inst Pasteur) 135A:47–53
Steiner T, McGarrity GJ, Phillips DM (1982) Cultivation and partial characterization of spiroplasmas in cell cultures. Infect Immun 35:296–304
Tully JG, Rose DL, Yunker CE, Cory J, Whitcomb RF, Williamson DL (1981) Helical mycoplasmas (spiroplasmas) inIxodes ticks. Science 212:1043–1045
Tully JG, Whitcomb RF (1981) The genusSpiroplasma. In: Starr M, Stolp H, Trüper HG, Balows A, Schlegel HG (eds) The prokaryotes, vol 2. New York: Springer-Verlag, pp 2271–2284
Tully JG, Whitcomb RF, Clark HF, Williamson DL (1977) Pathogenic mycoplasmas: cultivation and vertebrate pathogenicity of a new spiroplasma. Science 195:892–894
Tully JG, Whitcomb RF, Rose DL, Bové JM (1982)Spiroplasma mirum, a new species from rabbit ticks (Haemaphysalis leporispalustris). Int J Syst Bacteriol 32:92–100
Whitcomb RF (1980) The genusSpiroplasma. Annu Rev Microbiol 34:677–709
Whitcomb RF (1981) The biology of spiroplasmas. Annu Rev Entomol 26:397–425
Whitcomb RF (1983) Culture media for spiroplasmas. In: Razin S, Tully JG (eds) Methods in mycoplasmology, vol 1. New York: Academic Press, pp 147–158
Whitcomb RF, Clark TB, Tully JG, Chen TA, Bové JM (1983) Serological classification of spiroplasmas: current status. Yale J Biol Med 56:453–459
Whitcomb RF, Tully JG, Clark TB, Williamson DL, Bové JM (1982) Revised serological classification of spiroplasmas, new provisional groups, and recommendations for serotyping of isolates. Curr Microbiol 7:291–296
Williamson DL, Poulson DF (1979) Sex ratio organisms (spiroplasmas) ofDrosophila. In: Whitcomb RF, Tully JG (eds) The mycoplasmas, vol 3. New York: Academic Press, pp 175–208
Williamson DL, Steiner T, McGarrity GJ (1983)Spiroplasma taxonomy and identification of the sex ratio organism: can they be cultivated? Yale J Biol Med 56:583–592
Yunker CE, Cory J, Meibos H (1981) Continuous cell lines from embryonic tissues of ticks (Acari: Ixodidae). In Vitro 17:139–142
Yunker CE, Cory J, Meibos H (1984) Tick tissue and cell culture: applications to research in medical and veterinary acarology and vector-borne disease. In: Acarology VI, vol 2. Chichester UK: Ellis Harwood, pp 1082–1088
Yunker CE, Vaughn JL, Cory J (1967) Adaptation of an insect cell line (Grace'sAntheraea cells) to medium free of insect hemolymph. Science 155:1565–1566
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Yunker, C.E., Tully, J.G. & Cory, J. Arthropod cell lines in the isolation and propagation of tickborne spiroplasmas. Current Microbiology 15, 45–50 (1987). https://doi.org/10.1007/BF01577213
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DOI: https://doi.org/10.1007/BF01577213