Skip to main content
Acta Veterinaria Scandinavica logoLink to Acta Veterinaria Scandinavica
. 2021 Jan 23;27(1):23–32. doi: 10.1186/BF03548556

Improved Agar Plate Assays of Bovine Lysozyme and Haemolytic Complement Activity

Forbedrede agarplatemetoder til bestemmelse av bovint lysozym og hemolytisk complement aktivitet

Ø Lie 1,, M Syed 1, H Solbu 1
PMCID: PMC8189361  PMID: 3751812

Abstract

Bovine serum and colostral whey samples were examined for lysozyme and haemolytic complement activity, employing agar plate techniques. The tests were carried out in agarose gel containing Micrococcus lysodeikticus (for lysozyme), and antibody sensitized rabbit erythrocytes (for complement), respectively. The confirmation of lysozyme (E.C.3.2.1.17) — dependent lysis has been presented elsewhere (Lie & Syed 1986), while heat-inaotivation and antibody to C3 were used in the present study to confirm that the haemolytic activity was attributable to the complement cascade. Repeatability and sensitivity of the described tests were found to be superior to those of photometric procedures. Staining and preservation techniques were developed which extended the applicability of the assay, as they made reading of results independent of time and resulted in the plates being very suitable for photography and storage.

Full Text

The Full Text of this article is available as a PDF (1.9 MB).

References

  1. Barta O, Barta V. Haemolytic assay of bovine serum complement. J. immun. Methods. 1972;1:363–374. doi: 10.1016/0022-1759(72)90029-4. [DOI] [PubMed] [Google Scholar]
  2. Becker W A. Manual of Procedures in Quantitative Genetics. Pullman, Washington: Washington State University Press; 1967. p. 130. [Google Scholar]
  3. Bratlid D. A simple procedure for determination of bacteriolytic activity in biological fluids. Acta path, microbiol. scand. 1977;85:17–20. doi: 10.1111/j.1699-0463.1977.tb03605.x. [DOI] [PubMed] [Google Scholar]
  4. Jolies P, Jolies J. What’s new in lysozyme research? Always a model system, today as yesterday. Mol. cell. Biochem. 1984;63:165–189. doi: 10.1007/BF00285225. [DOI] [PubMed] [Google Scholar]
  5. Lachmann P J, Hobart M J. Complement technology. In: Weir D M, editor. Handbook of Experimental Immunology. Vol. 1 Immunochemistry. Oxford, London, Edinburgh, Melbourne: Blackwell Scientific Publications; 1979. pp. 5A.1–5A.23. [Google Scholar]
  6. Lie O. Genetic variation in serum lysozyme activity in cattle. Acta vet. scand. 1980;21:448–450. doi: 10.1186/BF03546877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Lie O, Solbu H. Evidence for a major gene regulating serum lysozyme activity in cattle. Z. Tierzucht. Ziichtbiol. 1983;100:134–138. doi: 10.1111/j.1439-0388.1983.tb00719.x. [DOI] [Google Scholar]
  8. Lie O, Syed M, Solbu H. The genetic influence on serum haemolytic complement levels in cattle. Anim. Blood Grps. biochem. Genet. 1983;14:51–57. doi: 10.1111/j.1365-2052.1983.tb01060.x. [DOI] [PubMed] [Google Scholar]
  9. Lie O, Solbu H, Syed M. A genetic association between bovine serum and colostrum lysozyme levels. Anim. Genet. 1986;17:39–45. doi: 10.1111/j.1365-2052.1986.tb03186.x. [DOI] [PubMed] [Google Scholar]
  10. Lie O, Syed M. Some properties of the lysozymes in serum and colostrum from cows with high and low lytic power against Micrococcus lysodeikticus. Anim. Genet. 1986;17:47–59. doi: 10.1111/j.1365-2052.1986.tb03187.x. [DOI] [PubMed] [Google Scholar]
  11. Mayer M M, Mayer M M. Complement and complement fixation. In: Kabat E A, editor. Experimental Immunochemistry. Springfield, Illinois: Charles Thomas; 1961. pp. 133–240. [Google Scholar]
  12. Osserman E F, Lawlor D P. Serum and urinary lysozyme (mura- minidase) in monocytic and monomyelocytic leukemia. J. exp. Med. 1966;124:921–951. doi: 10.1084/jem.124.5.921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Parry R M, Chandan R C, Shahani K M. A rapid and sensitive assay of muramidase. Proc. Soc. exp. Biol. (N. Y.) 1965;119:384–386. doi: 10.3181/00379727-119-30188. [DOI] [PubMed] [Google Scholar]
  14. Prieur D J, Camara V M. Inheritance of lysozyme deficiency in rabbits. J. Heredity. 1979;70:181–184. doi: 10.1093/oxfordjournals.jhered.a109230. [DOI] [PubMed] [Google Scholar]
  15. Rapp H J, Borsos T. Effects of low. ionic strength on immune haemolysis. J. Immunol. 1963;91:826–832. [PubMed] [Google Scholar]
  16. Rurangirwa F R, Tabel H, Losos G, Tizard I R. Hemolytic complement and C3 levels in Zebu Cattle infected with Trypanosoma congolense and Trypanosoma vivax and the effect of trypanocidal treatment. Infec. Immun. 1980;27:832–836. doi: 10.1128/IAI.27.3.832-836.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Virella G. Electrophoresis of lysozyme into Micrococcus-containing agarose gel: Quantitative and analytical applications. Clin, chim. Acta. 1977;75:107–115. doi: 10.1016/0009-8981(77)90505-8. [DOI] [PubMed] [Google Scholar]

Articles from Acta Veterinaria Scandinavica are provided here courtesy of BMC

RESOURCES