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PROFESSOR P. F. FRANKLAND AND MRS. G. C. FRANKLAND ON 
Bottles Nos. 1 and 3 (dilution ^,ooo obo ) yielded strong reactions both, -with diphenylamine and with 
sulphanilic acid, and both on microscopic examination were found to contain a large numher of 
very short, almost micrococcus-like bacilli. 
Bottle No. 2 (dilution no'y^ToFo)’ other hand, gave no reactions with either of the above 
reagents. 
Bottle No. 4 (dilution 0 ^ 75 - 5 - 5 ) gave no reactions. 
Bottle No. 5 (dilution 7 , 000 , 000 ) (examined on May 28, 1889) gave strong reactions with both 
reagents. 
Bottle No. 6 (dilution 1 , 000 . 000 ) (examined Alay 28, 1889) gave no reactions with the two reagents. 
Bottles Nos. 1 and 9 (dilution po 00 , 0 on ) gave strong reactions with both reagents. 
Bottle No. 8 (dilution 1 , 000 , 000 )? other hand, gave no reactions. 
On May 28tli, 1889, some of the bottles containing liquid still more dilute than 
T7 ooo,ooo were examined, but in every case with negative results. Thus :— 
Bottles Nos. 29, 30, 31, and 32 (dilution 50 . 000 , 000 )? all yielded negative results with both reagents. 
Thus of a number of bottles inoculated with about 1,00 0.~b~o0 original, some 
nitrified, Avhilst others did not, or, in other words, into some a nitrifying organism was 
introduced, whilst in others the inoculation failed to convey such an organism. 
On the other hand, all the bottles containing less than 1 , 00 0 .000 of the original 
failed to nitrify, no nitrifying organisms having been conveyed in these more highly 
attenuated inoculations. 
The most highly attenuated bottles which underwent nitrification being those 
which had received the dilution r,biro;b"oo? or bottles Nos. 1, 2, and 3, it was in these 
that the nitrifying organism in the greatest state of purity was to be expected. 
It was upon these bottles, therefore, that our further attention was concentrated. 
Of these bottles Nos. 1, 2, and 3, as already mentioned. Nos. 1 and 3 had nitrified 
strongly, and contained micro-organisms visible with the microscope, whilst No. 2 
had not nitrified. 
The contents of these three bottles were now submitted to investigation by means 
of cultivation-experiments. The results of these cultivation-experiments were highly 
remarkable. 
Thus on inoculating from each of these bottles into tubes of gelatine-peptone— 
(1) The tube from bottle No. 1, which had nitrified, never developed any growth 
whatever, although preserved for many months, indeed until the gelatine had become 
quite dried up. 
(2) The tube from bottle No. 2, which had not nitrified, exhibited already on the 
third day a growth, consisting of a white surface expansion and beaded in the depth, 
and causing no liquefaction of the gelatine. Under the microscope the growth was 
seen to consist of chains of small bacilli, with bright shining spores. 
(3) The tube from bottle No. 3, which had nitrified, also exhibited a growth. 
These results clearly showed that in tliese three bottles we had two distinct 
