1. The addition of manganese in a concentration as low as 1.8 x 10–5 M stimulates both respiration and germination markedly and eliminates the need for heat activation of spores respiring in inorganic salts-glucose.
2. Rapid germination and respiration of B. megatherium spores require a proper balance of ions in the nutrient medium. The repressive effect of a high phosphate concentration can be eliminated or balanced by chloride, or by other monovalent anions. Adenosine can relieve the necessity for chloride in such a system.
In two previous communications on the respiratory mechanism of the pneumococcus 1 the following observations were made. A washed suspension of various types and strains of pneumococcus uses up oxygen in the presence of glucose, sodium lactate, ethyl and propyl alcohols, and glycogen. The oxygen thus consumed is quantitatively converted into hydrogen peroxide as shown by analysis after a 10-20 minute reaction period. Later a discrepancy occurs, since the per cent of hydrogen peroxide found is less in proportion to the volume of oxygen Consumed. This discrepancy is caused by an intermediary reaction between hydrogen peroxide and pyruvic acid, these being formed simultaneously from glucose or lactic acid. In the case of lactic acid the reaction forms one molecule of acetic acid, one molecule of carbon dioxide, and one molecule of water. Besides these reaction products, we have isolated another acid of unknown constitution. The reactions involved in the formation of this acid we assume to be responsible for the excess of hydrogen peroxide found during the oxidation of lactic acid. Should lactic acid be quantitatively oxidized to hydrogen peroxide and pyruvic acid, they would react with one another at the time of their formation, and would be absent in the reaction mixture. However, since there is an excess of hydrogen peroxide, it must be assumed that the quantity of pyruvic acid formed is less than that of hydrogen peroxide, and the presence of this excess peroxide must be attributed to another source.
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