The assay of staphylococal delta-haemolysin with fish erythrocytes

• Main Author: Chao, Linpo
• Format: Thesis
• Language: English
• Published: ProQuest Dissertations & Theses 1976
• Subjects: Gadus morhua
• Online Access: http://theses.gla.ac.uk/72947/; http://theses.gla.ac.uk/72947/1/10647275.pdf

The purpose of this thesis was to investigate the suitability of cod and other fish erythrocytes for the assay of staphylococcal 5-haemolysin. The investigation adopted two broad approaches: (1) determining conditions for storage of fish erythrocytes, (2) the development of an assay system using fish erythrocytes as an indicator of haemolysis. The preservation of erythrocytes from cod (Gadus morhua), saithe (Pollachius virens) and mackerel (Scomber scombrus) was studied using diemthyl sulphoxide (DMSO) as a protective additive. Equal volumes of DMSO solution (15-30% in citrate dextrose saline) and citrated blood were mixed, rapidly frozen and stored in ampoules in liquid nitrogen. The optimum concentration of DMSO was 12.5% and erythrocyte recoveries of greater than 90% were obtained from all species after storage for up to five months. Larger quantities of blood were stored by removal of plasma from citrated blood prior to addition of DMSO solution, and by storage of pelleted frozen blood in aluminium canisters in liquid nitrogen. Maximum recoveries of washed intact erythrocytes required thawing of pellets in 12.5% DMSO solution and washing with citrate/dextrose/saline solutions containing decreasing concentrations of DMSO. Washed erythrocytes could be kept at 4