A simple and quickly performed colour test is described, in which organisms containing catechol 2:3‐oxygenase (meta‐pyrocatechase) produce from catechol a yellow coloured intermediate, α‐hydroxymuconic semialdehyde. The yellow colour is produced most consistently, rapidly and intensely by 2–4 day old cultures from 2·7–27 mM solutions of catechol. The purple‐brown colour produced by some organisms is discussed.
Using growth from nutrient agar plates, the test was applied to cultures of 239 Gram negative bacteria; all of the 64 cultures which gave a positive result had been in contact previously with aromatic substrates and many of them could oxidize phenol. The significance of a positive reaction is discussed briefly.
SUMMARY: Mixed cultures of bacteria grown in spent gas liquor readily oxidized phenol, o‐, m‐ and p‐cresol, catechol, 3‐methyl catechol, 4‐methyl catechol, resorcinol, 2‐methyl resorcinol, and 4‐methyl resorcinol. Quinol, pyrogallol and phloroglucinol were more resistant. The optimum temperature was 30° and the best pH range 6·5–7·8. Yeast extract and sterile sewage sludge both increased the rate of growth of organisms in liquor when the inoculum was small. Five phenol oxidizing organisms were isolated in pure culture. Copper in concentrations greater than 1 p/m inhibited both growth and phenol oxidation by one of these.
Mixed cultures grown in an ammonium thiocyanate medium originally inoculated with Thiobacillus thiocyanoxidans oxidized potassium thiocyanate and sodium thiosulphate. Chloride inhibited thiocyanate oxidation in concentrations above 5,000 p/m, although adaptation to 15,000 p/m was possible. Phenol inhibited thiocyanate oxidation in concentrations of 300 p/m or more. Mixed cultures grown on sodium thiosulphate oxidized sodium trithionate and tetrathionate, potassium pentathionate and hexa‐thionate, and potassium and ammonium thiocyanate
Manometric determinations of the 5 day biological oxygen demand of effluents after treatment showed good agreement with the values obtained by the conventional method, the manometric values being usually somewhat higher.
SUMMARY: A statistical experiment has shown that no significant proportion of the errors of determination of bacterial populations by the surface drop method could be attributed to the dropping technique employed. With both pure and mixed cultures the numbers of colonies/drop followed a Poisson distribution very closely.
It is suggested that in order to determine bacterial populations at a consistent level of accuracy a constant number of colonies should be counted rather than a constant number of drops. Whenever it is justified, the confidence limits for the results of counts should be quoted.
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