This is a report of attempts to compare the growth yields of various species of fastidious mycobacterium inch1ding human pathogens and non-pathogens in the conventional Dubos liquid medium and two simple media formulated recently; one is a medium containing 0.1% hyaluronic acid and 6.0% bovine serum albumin and the other is a semisyntheic medium made of umbilical cord extract supplemented with 10% sheep serum as a final concentration. All mycobacterial strains employed in experiments gave the heaviest growth yields in the hyaluronic acid-bovine serum albumin medium (HAS medium), among the three media. Dubos liquid medium seemed to be inferior to a medium made of umbilical cord extract (UCE medium) in supporting mycobacterial growth. There were three-to seven-fold increases in dry weight of the bacteria grown in the HAS medium as compared with those in the Dubos liquid medium. We also looked for the possible effect of bovine serum albumin (BSA)in the HAS medium on mycobacterial growth. As a result, we found that the amount of BSA in the HAS medium, ranging from zero to 6.0% in the medium, showed no substantial effect on the mycobacteria1 growth.
Comparative Study ; Culture Media/standards* ; Female ; Human ; Hyaluronic Acid/isolation & purification ; Hyaluronic Acid/pharmacology* ; Mycobacterium/growth & development* ; Pregnancy ; Tissue Extracts* ; Umbilical Cord*

Based on the biological activity of endotoxin, we propose a possible new method for detecting endotoxin using a pH- indication system of macrophage culture media. After RAW 264.7 macrophage cells were treated with lipopolysaccharide (LPS), the addition of fluorescein to the LPS-treated media reproductively reduced its absorption and emission spectra (it was a dose-dependent reduction). The advantages of this LPS- detection method were compared with the Limulus Amebocyte Lysate (LAL) test by using purified bacterial LPS (Salmonella minnessota, Escherichia coli, and Pseudomonas aeruginosa). Additionally, the absorption and fluorescence intensity of fluorescein, following treatment of RAW 264.7 cells with a high concentration of Staphylococcus aureus (Gram-positive, lysed bacteria), could not generally be detected by the LAL test, but they were found to be reduced, in a dose-response relationship, with this new system. The macrophage culture system-method might be a good supplement to the LAL assay for detection of LPS, Gram-negative and Gram-positive bacteria.
Animals ; Biological Assay/*methods/standards ; Cells, Cultured ; Comparative Study ; *Contrast Media ; Culture Media/chemistry ; Endotoxins/*analysis ; *Fluorescein ; Hydrogen-Ion Concentration ; Limulus Test ; Lipopolysaccharides/analysis ; Macrophages/*chemistry ; Mice ; Research Support, Non-U.S. Gov't