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Improved Agar Plate Assays of Bovine Lysozyme and Haemolytic Complement Activity

Forbedrede agarplatemetoder til bestemmelse av bovint lysozym og hemolytisk complement aktivitet

Acta Veterinaria Scandinavica volume 27, pages 23–32 (1986)Cite this article

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.

Sammendrag

Bovint serum og kolostrum ble undersøkt for lysozym- og hemolytisk complement-aktivitet ved hjelp av lysoplateteknikker. Testene ble utført i agaorse-gel inneholdende Micrococcus lysodeikticus (for lysozym) og antistoff — sensiterte kaninerytrocytter (for complement). Bekreftelsen på lysozym — spesifikk lysis er utført ved biokjemiske analyser i et annet studium (Lie et al. 1986) mens varmeinaktivering og spesifikke anti-C3-antistoffer ble anvendt i dette arbeidet for å bekrefte complement-avhengig hemolyse. Reproduserbarhet, sensitivitet og kapasitet på de beskrevne platemetoder ble funnet å være betydelig bedre enn for fotometriske tester. Dessuten ble fargings-/preserveringsteknikker utviklet, noe som øker anvendbarheten av testene i det avlesninger blir tidsuavhengige og platene velegnet for fotografering og lagring.

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Authors and Affiliations

  1. National Veterinary Institute, P. O. Box 8156, Dep., N-0033, Oslo 1, Norway

    Ø. Lie, M. Syed & H. Solbu

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  1. Ø. Lie
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  2. M. Syed
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  3. H. Solbu
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Lie, Ø., Syed, M. & Solbu, H. Improved Agar Plate Assays of Bovine Lysozyme and Haemolytic Complement Activity. Acta Vet Scand 27, 23–32 (1986). https://doi.org/10.1186/BF03548556

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Acta Veterinaria Scandinavica

ISSN: 1751-0147