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Observations on the maintenance of Trichomonas vaginalis and Trichomonas foetus; the effects of cortisone and agar on enhancement of severity of subcutaneous lesions in mice

Published online by Cambridge University Press:  06 April 2009

L. Jeffries  and
Margaret Harris
L. Jeffries
Affiliation:
Walton Oaks Experimental Station, Tad worth, Surrey
Margaret Harris
Affiliation:
Walton Oaks Experimental Station, Tad worth, Surrey
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Methods are described for the freezing of trichomonads, suspended in a liverinfusion broth, to – 20 °C and – 70 °C, and storage at these temperatures. After 9 months’ storage at – 70 °C, with dimethyl sulphoxide, six strains of T. vaginalis and three strains of T. foetus were alive; three trichomonas strains frozen earlier were also alive after 13 months. Four of the T. vaginalis strains survived equally well for 6 months at – 70 °C, in the presence of either dimethyl sulphoxide or glycerol.

Although trichomonads may be maintained by subculture in a modified Diamond medium, storage at – 70 °C is more economical and less tedious; – 20 °C is not a satisfactory storage temperature for T. vaginalis or T. foetus, even in the presence of dimethyl sulphoxide or glycerol.

The severity of subcutaneous lesions in mice produced by six freshly isolated strains of T. vaginalis, and two out of three stock culture strains of T. foetus, was enhanced by either the addition of agar to the suspending medium or administration of cortisone to the mice. One strain of T. foetus produced lesions only in the presence of agar, cortisone, on the dose schedule used, being ineffective.

After storage at – 70 °C or cultivation in agar medium for 6 months, the virulence of trichomonads of either species was little affected, and lesions in mice were enhanced by agar in a similar degree to that found originally.

For enhancing the severity of subcutaneous lesions in mice the agar method, involving only one injection, is simpler than the repeated administration of cortisone to infected mice; agar was shown to be more effective than cortisone with small infecting doses of one strain of T. vaginalis quantitatively tested.

The subcutaneous injection of mice with trichomonads suspended in a liver-infusion broth containing 0·06% agar regularly produces lesions of the degree of severity required for the evaluation of compounds for therapeutic activity.

We express our gratitude to Mr S. A. Price for constructive criticism and helpful suggestions during the preparation of the manuscript. Dr S. R. M. Bushby kindly provided details of the dimethyl sulphoxide method and apparatus in use in his laboratory for the low-temperature preservation of tumour cells and trichomonads. For supplying the trichomonas strains we are grateful to Dr L. P. Joyner, Miss M. Moore-Tucker and Mr C. S. Ledwich. We thank Mr W. A. Freeman for advice on the composition of the inoculum medium.

Type
Research Article
Information
Parasitology , Volume 57 , Issue 2 , May 1967 , pp. 321 - 334
Copyright
Copyright © Cambridge University Press 1967

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References

REFERENCES

Anon (1964). Metronidazole, Br. med. J. 2, 803–4.CrossRefGoogle Scholar
Diamond, L. S. (1957). The establishment of various trichomonads of animals and man in axenic cultures. J. Parasit. 43, 488–90.CrossRefGoogle ScholarPubMed
Dougherty, R. M. (1962). The use of dimethyl sulphoxide for preservation of tissue culture cells by freezing. Nature, Lond. 193, 550–2.CrossRefGoogle ScholarPubMed
Fitzgerald, P. R. & Levine, N. D. (1957). The effects of storage temperature and other factors on survival of Trichomonas foetus at freezing temperatures. J. Protozool. 4, Suppl. 5.Google Scholar
Gledhill, A. W. & Rees, R. J. W. (1952). Spontaneous enterococcal disease in mice and its enhancement by cortisone. Br. J. exp. Path. 33, 183–9.Google ScholarPubMed
Honigberg, B. M. (1961). Comparative pathogenicity of Trichomonas vaginalis and Trichomonas gallinae to mice. 1. Gross pathology, quantitative evaluation of virulence, and some factors affecting pathogenicity. J. Parasit. 47, 545–71.CrossRefGoogle Scholar
Ivey, M. H. (1961). Growth characteristics of clones of Trichomonas vaginalis in solid medium. J. Parasit. 47, 539–44.CrossRefGoogle ScholarPubMed
Johnson, G. & Trussell, R. E. (1943). Experimental basis for the chemotherapy of Trichomonas vaginalis infestations. 1. Proc. Soc. exp. Biol., Med. 54, 245–9.CrossRefGoogle Scholar
Levine, N. D., Andersen, F. L., Losch, M. B., Notzold, R. A. & Mehra, K. N. (1962) Survival of Tritrichomonas foetus stored at – 28 and – 95 °C after freezing in the presence of glycerol. J. Protozool. 9, 347–50.CrossRefGoogle ScholarPubMed
Levine, N. D. & Marquardt, W. C. (1955). The effect of glycerol and related compounds on survival of Tritrichomonas foetus at freezing temperatures. J. Protozool. 2, 100–7.CrossRefGoogle Scholar
Lindgren, R. D. & Ivey, M. H. (1964). The effect of cultivation and freezing on the virulence of Trichomonas vaginalis for mice. J. Parasit. 50, 226–9.CrossRefGoogle ScholarPubMed
Lovelock, J. E. & Bishop, M. W. H. (1959). Prevention of freezing damage to living cells by dimethyl sulphoxide. Nature, Lond. 183, 1394–5.CrossRefGoogle ScholarPubMed
Lynch, J. E., Holley, E. C. & Margison, J. E. (1955). Studies on the use of the mouse as a laboratory animal for the evaluation of antitrichomonal agents. Antibiotics Chemother. 5, 508–14.Google ScholarPubMed
McEntegart, M. G. (1954). The maintenance of stock strains of trichomonads by freezing. J. Hyg. Camb. 52, 545–50.Google ScholarPubMed
McEntegart, M. G. (1959). Prolonged survival of Trichomonas vaginalis at – 79 °C. Nature, Lond. 183, 270–1.CrossRefGoogle Scholar
Miller, J. K. & Hedberg, M. (1965). Effects of cortisone on susceptibility of mice to Listeria monocytogenes. Am. J. clin. Path. 43, 248–50.CrossRefGoogle ScholarPubMed
Olitzki, A. L. (1957). The infection promoting substances (a review), Bull. Res. Coun. Israel 6 E, 193–222.Google Scholar
Polge, C. & Lovelock, J. E. (1952). Preservation of bull semen at –79 °C., Vet. Rec. 64, 396–7.Google Scholar
Porterfield, J. S. & Ashwood-Smith, M. J. (1962). Preservation of cells in tissue culture by glycerol and dimethyl sulphoxide. Nature, Lond. 193, 548–50.CrossRefGoogle ScholarPubMed
Richardson, U. F. & Kendall, S. B. (1957). Veterinary Protozoology, pp. 238–9. Edinburgh: Oliver and Boyd.Google Scholar
Robinson, S. C. (1962). Trichomonal vaginitis resistant to metronidazole. Can. med. Ass. J. 86, 665.Google Scholar
Ryley, J. F. & Stacey, G. T. (1963). Experimental approaches to the chemotherapy of trichomoniasis. Parasitology 53, 303–20.CrossRefGoogle Scholar
Schnitzer, R. J., Kelly, D. R. & Leiwant, B. (1950). Experimental studies on trichomoniasis. 1. The pathogenicity of trichomonad species for mice. J. Parasit. 36, 343–9.CrossRefGoogle ScholarPubMed
Squires, S. & McFadzean, J. A. (1962). Strain sensitivity of Trichomonas vaginalis to Metronidazole. Br. J. vener. Dis. 38, 218–19.Google ScholarPubMed
Watt, L. (1965). Trichomoniasis. Practitioner 195, 613–9.Google ScholarPubMed
Watt, L. & Jennison, R. F. (1962). Metronidazole treatment of trichomoniasis in the female. Br. med. J. 1, 276–9.CrossRefGoogle ScholarPubMed