Date of Award:
5-1-1975
Document Type:
Dissertation
Degree Name:
Doctor of Philosophy (PhD)
Department:
Biology
Department name when degree awarded
Bacteriology (Bacteriology and Public Health)
Committee Chair(s)
Rex S. Spendlove
Committee
Rex S. Spendlove
Committee
Paul B. Carter
Committee
Ross A. Smart
Committee
James L. Shupe
Committee
LeGrande C. Ellis
Abstract
The purposes of this investigation were to develop an improved method for the detection of anti-mycoplasmal antibody and the partial characterization of the effect of Mycoplasma arthritidis on lysosomal enzyme release from cells in culture. A new, rapid, sensitive and specific serologic test for the detection of anti-mycoplasmal antibody is described. This technique is based on the agglutination of acridine orange stained, whole mycoplasmas by homologous antiserum. The antiserum-aggregated, stained antigen is observed using the incident illumination fluorescence microscope. Titer of the antiserum is taken as the reciprocal of the highest antiserum dilution giving agglutination. Autoagglutination was a serious problem with certain mycoplasma species. However, the problem was overcome by using 10 percent fetal calf serum as the antiserum diluent. Optimal conditions for the fluorescent mycoplasma agglutination (FMA) test are: (1) Stain with acridine orange and harvest the antigen after 36-48 hours of incubation. (2) Incubate the stained cells with diluted specific antibody at 42 C for one hour. (3) Centrifuge the mixture at 700 X G for 10 minutes. (4) Place a drop of the antigenantibody mixture on a glass microscope slide and observe for agglutination. This test has been adapted to the microtiter system. The effect of M. arthritidis on lysosomal enzyme release from cells growing in vitro was assessed by monitoring the enzyme activity in the cell maintenance medium. It was found that the lysosomal enzyme, beta-glucuronidase, was released earlier and in larger amounts in the infected cell cultures. The cytoplasmic marker, lactate dehydrogenase (LDH), was not released in significantly larger amounts in infected cells. Variation in numbers of lysosomes was determined by vitally staining infected and non-infected cells with acridine orange. Infected cells always contained fewer lysosomes than did uninfected cells.
Recommended Citation
Foster, Leland George, "An Improved Serologic Test for Mycoplasmas and the Effect of Mycoplasma arthritidis on the Release of Lysosomal Enzymes from Cell Cultures" (1975). Biology. 410.
https://digitalcommons.usu.edu/etd_biology/410
Copyright for this work is retained by the student. If you have any questions regarding the inclusion of this work in the Digital Commons, please email us at .