Hostname: page-component-586b7cd67f-t7czq Total loading time: 0 Render date: 2024-11-20T11:33:15.749Z Has data issue: false hasContentIssue false
  • English
  • Français

Rapid membrane filtration epifluorescent microscopic technique for the direct enumeration of somatic cells in fresh and formalin-preserved milk

Published online by Cambridge University Press:  01 June 2009

Graham L. Pettipher  and
Ubaldina M. Rodrigues
Ubaldina M. Rodrigues
Affiliation:
National Institute for Research in Dairying, Shinfield, Reading RG2 9AT
Get access Rights & Permissions [Opens in a new window]

Summary

A rapid method for the direct enumeration of somatic cells in fresh and formalin-preserved raw milk is described. Heat treatment at 80°C for 10 mn fixed somatic cells sufficiently to prevent lysis and subsequent dilution allowed the equivalent of at least 0.5 ml milk to be filtered through a 1.0 μm pore size Nuclepore membrane filter. The somatic cells were concentrated on the membrane and, after staining with acridine orange, fluoresced green, yellow or orange under an epifluorescent microscope. Three different types of cell could be distinguished on the basis of nuclear structure and cell size. These were monocytes, polymorphonuclear leucocytes and the larger epithelial and/or secretory cells. For both fresh and preserved milks the count of somatic cells on the membrane correlated well (r ≽ 0·93) with the Coulter count. Differences between counts obtained by different operators were not significant. The technique is rapid, taking about 20min, and is suitable for milks containing between 2 × 104 and 1 × 107 somatic cells/ml.

Type
Original Articles
Information
Journal of Dairy Research , Volume 48 , Issue 2 , June 1981 , pp. 239 - 246
Copyright
Copyright © Proprietors of Journal of Dairy Research 1981

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

American Public Health Association (1978). Standard methods for the examination of dairy products, 14th Edn. New York: American Public Health Association, Inc.Google Scholar
Bossuyt, R. (1978). Milchwissenschaft 33, 11–13.Google Scholar
Bramley, A. J. (1978). British Veterinary Journal 134. 146–151.CrossRefGoogle Scholar
Bramel, R. D., Schultz, L. H., Gabler, F. R. & Peters, J. (1977). Journal of Food protection 40, 32–38.CrossRefGoogle Scholar
Duitschaever, C. L. & Leggatt, A. G. (1967). Stain Technology 42, 183–194.CrossRefGoogle Scholar
Gordon, W. A., Morris, H. A. & Packard, V. (1980). Journal of Food Protection 43, 58 64, 75.CrossRefGoogle Scholar
International Dairy Federation (1979). Bulletin International Dairy Federation (Document no. 114).Google Scholar
Lee, C. S., Wooding, F. B. P. & Kemp, P. (1980). Journal of Dairy Research 47, 39–50.CrossRefGoogle Scholar
Luedecke, L. Q., Forster, T. L. & Ashworth, U. S. (1967). Journal of Dairy Science 50, 1592–1596.CrossRefGoogle Scholar
Marshall, R. T. (1978). Bulletin, International Dairy Federation (Document no. 108), pp. 5–9.Google Scholar
Pettipher, G. L., Mansell, R., McKinnon, C. H. & Cousins, C. M. (1980). Applied and Environmental Microbiology 39, 423–429.CrossRefGoogle Scholar
Pettipher, G. L. &, Rodrigues, U. M. (1981). Journal of Applied Bacteriology 50, 157–166.CrossRefGoogle Scholar
Thompson, D. R., Packard, V. S. & Ginn, R. E. (1976). Journal of Milk and Food Technology 39, 854–858.CrossRefGoogle Scholar
Von Bertalanffy, L. & Bickis, I. (1956). Journal of Histochemistry and Cytochemistry 4, 481–493.CrossRefGoogle Scholar