34 
THE GARDENING WORLD. 
January 14, 1905. 
had no inducement or interest for exerting their former func¬ 
tions, and, getting all the necessary food of this particular 
nature without any trouble on their part, they gradually lost 
the power of fixing the nitrogen of the atmosphere. Then, 
when applied to the soil in which any particular crop was to be 
reared, they gave no results as compared with other soil which 
had not been inoculated. 
Dr. Moore further continued his experiments and found that 
by gradually reducing the amount of nitrogen in the food 
material in which he cultivated the organisms that it was pos¬ 
sible to increase the nitrogen-fixing power of the bacteria. 
When properly manipulated it was found that these organisms, 
originally obtained from tlie roots of Clover, Beans, 01 otliei 
leguminous plants, could be made much more active in fixing 
nitrogen than when left to their own resources in a state of 
nature. , , 
He had his discovery patented in the United States, but 
with unprecedented liberality gave the results of his researches 
free for use by the farmers in the United States. The process 
followed by this investigator has been admirably summed up in 
his patent-application, from which we glean the particulars of 
his method of setting to work. His invention has for its object 
the production of more highly effective organisms and their 
distribution in a form preventing deterioration and which may 
be readily applied to cultures, whether in garden or field. 
The plants from which he obtained the organisms upon 
which lie set to work were Clovers, Cow Peas, Beans, and other 
leguminous plants employed in American agriculture. the 
tubercles were first obtained from the said plants, thoroughly 
washed and surface sterilised by the ordinary means well 
known to the chemist in the laboratory. The contents of the 
nodules were then cut out under sterile conditions, the object 
being to prevent the access of other and undesirable organisms 
and to maintain these special cultures pure and clean. 
This matter was then placed in water containing about 1 per 
cent, commercial agar-agar, 1 per cent, maltose sugar or cane 
sucrar .02 to .05 per cent, magnesium sulphate, and about 0.1 
per cent, of monobasic potassium phosphate. This solution 
was sterilised according to ordinary bacteriological processes. 
The solution provided as food for the bacteria differed fiom 
ordinary mixtures employed for similar purposes m the absence 
of combined nitrogen. , , , 
The above percentages were not intended to be absolute, as 
they might vary within given limits and still give excel ent 
results They were, however, what the doctor considered o 
be the best for giving the desired results. The organism m 
hand continues to multiply so long as the food materials m 
solution are not exhausted. 
The first stage of this process is to separate the orgams 
belonging to any of the leguminous plants m question and to 
cultivate it pure, so that it may benefit the particular crop to 
Which it is to be applied. The operator’s care was directed 
to the keeping out of mould and other organisms which would 
defeat the object he had in view, so that it may be interesting 
to wardeners to remember that even the smallest member of 
the & vegetable kingdom may have its own enemy or enemies 
These organisms being entirely without chlorophyll or lea 
wreen, it is immaterial whether they are cultivated m light or 
darkness. The temperature, however, best suited to then de¬ 
velopment must range between 68 deg. and 86 deg. X ■ 
When these pure cultures aVe once obtained they may 
multiplied indefinitely, and immediately used or kept tor some 
weeks if necessary by methods pursued by those versed m bac¬ 
teriology. If the intention is to multiply them for immediate 
use, a quantity is put in water containing about 1 per cen . o 
cane sugar or maltose (the latter being best), about, .0 . to 
per cent, of magnesium sulphate, and 0.1 jier cent, o mono 
basic potassium phosphate or other ingredients containing 
these necessary elements of plant food. . . 
The organisms multiply very rapidly in this liquid and. a- 
will be noted, the medium contains no nitrogen in combination, 
so that the organisms have to complete tlieir bill of fare >v 
collect inw the free nitrogen of the atmosphere and combining 
it for their own needs. They do avail themselves ot atmo¬ 
spheric nitrogen that may be dissolved m the liquid in which 
they are cultivated, but practically that is also free or uncom¬ 
bined with any other element. In the course of a few days 
the liquid assumes a milky appearance owing to the immense 
number of bacteria present in it. 
This, then, is the stage at which the bacterial culture can 
be turned to practical use to which a correspondent referred 
on page 1038 of our last volume. The liquid containing the 
organisms may be sprinkled directly upon the soil it is in¬ 
tended to inoculate or upon seeds about to be sown. Where 
the culture is to be of any considerable extent, a. very ready 
method would be to place the seeds in the solution, which 
would make certain that the surface of each would be in¬ 
oculated with the germ, and therefore ensure an equal distribu¬ 
tion in the land where such sowing is made. 
If the intention is to preserve any particular organism till 
some future period cotton or some similar material is dipped 
into the liquid containing the bacteria, and the material is then 
thoroughly dried by methods employed by chemists. This has 
the effect of causing the organism to assume that form in 
which it can rest until the conditions are again favourable to 
further growth and multiplication. In this dry condition any 
of the desired bacteria may be kept for an indefinite or reason¬ 
able period, so that it can be transported by post or rail to any 
destination. 
When this cotton impregnated with bacteria reaches its des¬ 
tination, the process is simply repeated as above mentioned by 
dropping the cotton into a solution containing the desired 
elements of food required by the organism. In this operation 
cleanliness only is necessary. When the operator is preparing 
a culture for immediate use in the garden or field, all that he 
need do is to use clean vessels with ordinary well or rain water 
in which the food material may be placed and kept at a suffi¬ 
cient temperature for twelve to forty-eight hours until the 
bacteria have so multiplied as to make the preparation ready 
for use. If it is desired at this stage to stimulate the mul¬ 
tiplication of these organisms, it can be done by adding 1 per 
cent, of phosphate of ammonia to the solution. After the 
addition of the stimulant the culture should not be continued 
longer than twelve to forty-eight hours. This is only intended 
for the organisms to be used for seeding purposes immediately 
and not -or preservation. 
Dr. Moore now considers that the first attempt to fix the free 
nitrogen of the atmosphere by practicable means has been suc¬ 
cessfully made. The use of the organisms can only be of direct 
benefit to crops belonging to the Pea family, but any other 
crop can benefit by the residual nitrogen left in the soil in the 
form of roots' and other remains of the previous crop. 
The importance of the above discovery is due to the fact that 
the natural deposits of nitrate in Chili are not likely to con¬ 
tinue for ever, and according to report, the beds are veiy 
nearly exhausted. If all the known deposits of nitrogenous 
matter in an available form were to become exhausted, agri¬ 
culture, and horticulture, for the matter of that, would suffer 
immensely throughout the world for lack of one of the most 
important artificial fertilisers that have to be applied by man 
in his cultural operations. We may state, however, that the 
above is one likely source of nitrate in the future, because it is 
likely to remain true that when one material supplying this 
valuable element becomes exhausted science keeps on discover¬ 
ing some further source of supply. 
Dr. Bruno Terne, another great chemist in the United 
States, has been giving his attention for many years past to 
other likelv sources of combined nitrogen, and embodies the 
result of some of his own researches and experiments carried 
out by others under his direction in a paper before the Chemi¬ 
cal Section, and which now appears in the "Journal of the 
Franklin Institute ” for December. His attention was specially 
directed to the utilisation of the by-products of the coke in¬ 
dustry, and he relates some of his experiments in the separa¬ 
tion of sulphate of ammonia from the ammoniacal liquors of 
the gasworks which are usually a neglected waste product. 
About thirteen years ago not a pound of sulphate of am¬ 
monia was producted from this waste material, but now the 
salt extends to 25,000 to 30,000 tons per annum. He thinks 
that this will continue until the home production of that fei- 
