A word upon the natural distribution of these nitrifying bacteria before we leave them. They belong to the soil, river water, and sewage. They are also said to be frequently present in well water. From some experiments at Rothamsted it appears that the organisms occur mostly in the first twelve inches, and in subsoils of clay down to three or four feet. In sandy soils nitrification may probably occur at a greater depth. These facts should be borne in mind when arranging for the purification of sewage by intermittent filtration. We have now given some consideration to the chief events in the life-cycle of nature depicted in the table. There is but one further process in which bacteria play a part, and which requires some mention. It will have been noticed that at certain stages in the cycle there is more or less appreciable "loss" of free nitrogen. In the process of decomposition brought about by the denitrifying bacteria, a very considerable portion of the nitrogen is dissipated into the air in the form of a free gas. This is the last stage of all proteid decomposition, so that wherever putrefaction is going on there is a continual "loss" of an element essential to life. Thus it would appear at first sight that the sum-total of nitrogen food must be diminishing. But there are other ways also in which nitrogen is being set free. In the ordinary processes of vegetation there is a gradual draining of the soil and a passing of nitrogen into the sea; the products of decomposition pass from the soil by this drainage, and are "lost" as far as the soil is concerned. Many of the methods of sewage disposal are in reality depriving the land of the return of nitrogen which is its necessity. Again, nitrogen is freed in explosions of gunpowder, nitroglycerine, and dynamite, for whatever purpose they are used. Hence the great putrefactive "loss" of nitrogen, with its subsidiary losses, contributes to reduce this essential element of all life, and if there were no method of bringing it back again to the soil, it would seem that plant life, and therefore animal life, would speedily terminate. It is at this juncture, and to perform this vital function, that the _nitrogen-fixing bacteria_ play their wonderful part: they bring back the free nitrogen and fix it in the soil. Excepting a small quantity of combined nitrogen coming down in rain and in minor aqueous deposits from the atmosphere, the great source of the nitrogen of vegetation is the store in the soil and subsoil, whether derived from previous accumulations or from recent supplies by manure.