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Optimization of culture conditions for the maintenance of Onchocerca gutturosa adult worms in vitro

Published online by Cambridge University Press:  05 June 2009

Simon Townson ,
C. Connelly  and
R. Muller
Simon Townson
Affiliation:
CAB-International Institute of Parasitology, 395 A Hatfield Road, St. Albans, Herts, AL40XQ, England
C. Connelly
Affiliation:
CAB-International Institute of Parasitology, 395 A Hatfield Road, St. Albans, Herts, AL40XQ, England
R. Muller
Affiliation:
CAB-International Institute of Parasitology, 395 A Hatfield Road, St. Albans, Herts, AL40XQ, England
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Abstract

A series of experiments examined the effects of various media, serum supplements, gas phases and the incorporation of mammalian cell feeder layers on the survival on Onchocerca gutturosa adult worms in vitro. The survival of male worms was poor in all media tested that were not supplemented with inactivated foetal calf serum (IFCS), with improved but variable survival in media supplemented with 10–30% IFCS. Using a cell-free system in an atmosphere of 5% CO2in air, good results were obtained in medium NCTC 135 + 10% IFCS (median survival time 39 days, range 25–41). Marginally better survival was obtained with the same medium in an atmosphere of 95% N2/5% CO2 (medium 45 days, range 25–56)and with a 1:1 mixture of media NCTC 135 and IMDM+10% IFCS(medium 38 days, range 38–51). Survival was enhanced in culture systems which incorporated bovine kindney (MDBK) cells, bovine trachea (EBTR) cells and monkey kidney (LLCMK2) cells. Exceptionally long survival was obtained using medium MEM+10% IFCS+LLCMK2 cells under a gas phase of 5% CO2in air, in which male worms survived from approximately 6 to over 7 months. Under similar conditions, female worms were also maintained for periods of up to 6 months and 5 out of 18 specimens released microfilariae into the culture system. The long-term culture described in this study will be useful for basic biochemical, chemotherapeutic and immunological studies in vitro.

Keywords

Onchocerca gutturosain vitro cultureadults
Type
Research Papers
Information
Journal of Helminthology , Volume 60 , Issue 4 , December 1986 , pp. 323 - 330
Copyright
Copyright © Cambridge University Press 1986

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References

REFERENCES

chen, S. N. & Howells, R. E. (1979). The in vitro cultivation of the infective larvae and the early mammalian stages of the filarial worm Brugia Pahangi. Annals of Tropical Medicine and Parasitology, 73, 473–486.CrossRefGoogle ScholarPubMed
Eichler, D. A. (1973) Studies on Onchocerca gutturosa (Newumann, 1910)and its development in Simulium ornatum (Meigen, 1818). 3. Factors affecting the development of the parasite in its vector. Journal of Helminthology, 47, 73–88.CrossRefGoogle ScholarPubMed
Engelbrecht, F. & Schulz-key, H. (1984) Observations on adult Onchocerca volvulus maintained in vitro. Transactions of the Royal Society of Tropical Medicine and Hygiene, 78, 212–215.CrossRefGoogle ScholarPubMed
Franke, E. D. & weinstein, P. P. (1984) In vitro cultivation of Dipetalonema viteae third-stage larvae: effect of the gas phase. Journal of Parasitology, 70, 493–498.CrossRefGoogle ScholarPubMed
Fuglsang, H. & Anderson, J. (1976) Seasonal variation in the concentration of Onchocerca volvulus microfilariae in the skin? Tropenmedizin und Parasitologie, 27, 365–369.Google ScholarPubMed
Hashinguchi, Y., Kawabata, M., Tanaka, I., Okazawa, T., Flores, C. & Recinos, M. M. (1981) Seasonal variation in the microfilarial skin density of Onchocerca volvulus and in the biting activity of Simulium spp. in Guatemala. Transactions of the Royal Society of Tropical Medicine and Hygiene, 75, 839–845.CrossRefGoogle Scholar
Kozek, W. J. & Flgueroa, H. (1982) Attempts to establish O. volulus infection in primates and small laboratory animals. Acta Tropica, 39, 317–321.Google Scholar
Lok, J. B., Pollack, R. J., Cupp, E. W., Bernardo, M. J., Donnelly, J. J. & Alblez, E. J. (1984). Development of Onchocerca lienails and O. volvulus from third to fourth larval stage in vitro. Tropenmedizin und Parasitologie, 35, 209–211.Google Scholar
Mak, J. W., Lim, P. K. C., Sim, B. K. L. & Liew, L. M. (1983). Brugia malayi and B. Pahangi: cultivation in vitro of infective larvae to the fourth and fifth stages. Experimental Parasitology, 55, 243–248.CrossRefGoogle Scholar
Miegeville, M., Bouillard, C., Marjolet, M., Vermeil, C. & Avranche, P. (1981) Nouvelle contribution á l'étude de Dipetalonema viteae: maintien en survie des adultes in vitro. Bullein de la Société de Pathologie Exotique, 74, 207–216.Google ScholarPubMed
Prince, A. M., Alblez, E. J. & Van Den Ende, M. C. (1985). Onchocerca volvulus: transplantation of adult worms into chimpanzees. Tropical Medicine and Parasitology, 36, 105–108.Google ScholarPubMed
Schiller, E. L., Turner, V. M., Figueroa-Marroquin, H. & D'antonio, R. (1979) The cryopreservation and in vitro cultivation of larval Onchocerca volvulus. American Journal of Tropical Medicine and Hygiene, 28, 997–1009.CrossRefGoogle ScholarPubMed
Suswillo, R. R., Nelson, G. S., Muller, R., Mcgreevy, P. B., Duke, B. O. L. & Denham, D. A. (1977) Attempts to infect jirds(Meriones unguiculatus) with Wuchereria bancrofit, Onchocerca volvulus, Loa loa loa and Mansonella ozzardi. Journal of Helminthology, 5, 132–134.CrossRefGoogle Scholar
Tanner, M. (1981) Dipetalonema viteae (Filarioidea): development of the infective larvae in vitro. Acta Tropica, 38, 241–249.Google ScholarPubMed
Taylor, A. E. R. (1960) Maintenance of filarial worms in vitro. Experimental Parasitology, 9, 113–120.CrossRefGoogle ScholarPubMed
Townson, S., Blanco, A. E. & Owen, D., (1981a) Attempts to infect small laboratory animals with the infective larvae of Onchocerca lienalis. Journal of Helminthology, 55, 247–249.CrossRefGoogle ScholarPubMed
Townson, S., El Sinnary, K. & Bianco, A. E. (1981b) Transplantation of Onchocerca into mice. Transactions of the Royal Society of Tropical Medicine and Hygiene, 75, 889.Google Scholar
Townson, S., Connelly, C. (1986) Long-term survival Onchocerca gutturosa adult worms in vitro. Tropical Medicine and Parasitology, 37, 94.Google Scholar
Townson, S., Connelly, C. & Ham, P. J. (1986) Cultivation of Onchocerca lienalis developing larvae in vitro. Tropical Medicine and Parasitoligy, 37, 94.Google Scholar
Van Den Ende, M. C., Alblez, E. J. & Dennis, E. A. (1981) Transplantation of adult Onchocerca volvulus into chimpanzees. Transactions of the Royal Society of Tropical Medicine and Hygiene, 75, 310–311.CrossRefGoogle ScholarPubMed
Weinstein, P. P. & Sawyer, T. K. (1961) Survival of Dirofilaria uniformis in vitro and their production of microfilariae. Journal of Parasitology, 47, Suppl. 23.Google Scholar
Wenk, P., Illgen, B. & Seitz, H. M. (1978) In vitro Versuche mit Litomosoides carinii(Nematoda: Filarioidea). Zeischrift für Parasitenkunde, 55, 63–72.CrossRefGoogle Scholar
WHO (1983) Onchocerciasis Chemotherapy Project. Report of a scientific working group on the biochemistry of filarial and allied parasites. Publ. by the World Health Organization, OCP/OCT/83.1.Google Scholar
Wong, M. M., Wada, M., Fidel, J., Krighton, R. & Diling, G. (1980) In vitro cultures of infective stage larvae of Dirofilaria immitis and Brugia pahangi. Abstracts of 10th International Congress on Tropical Medicine and Malaria, Manila, Philippines, November 9–15, pp. 111–112.Google Scholar