The biochemical decomposition of nitrogenous substances in soils


Material Information

The biochemical decomposition of nitrogenous substances in soils
Series Title:
Bulletin / Hawaii Agricultural Experiment Station ;
Physical Description:
25 p. : ; 23 cm.
Kelley, W. P ( Walter Pearson ), b. 1878
U.S. G.P.O.
Place of Publication:
Washington, D.C
Publication Date:


Subjects / Keywords:
Soils -- Analysis   ( lcsh )
Soils -- Nitrogen content   ( lcsh )
federal government publication   ( marcgt )
bibliography   ( marcgt )
non-fiction   ( marcgt )


Includes bibliographical references.
Statement of Responsibility:
by W.P. Kelley.

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Source Institution:
University of Florida
Rights Management:
All applicable rights reserved by the source institution and holding location.
Resource Identifier:
aleph - 029613128
oclc - 16324195
lcc - S52 .E1 no. 32-50
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Full Text


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WUSTOATIS Aaronomist in Charv

Bulletin No. 396


IN ... ..

vann THE Bavenvuffox or

W. P. KELLEY,1 ChemiSt.
D. T. FULLAWAY, Entuafoogist.
W. T. MCGEORGE, Chemist.
ALICE R. THOMPSON, Assistant Chemist.
V. S. HOLT, Assistant Horticulturist.
C. A. SAnR, Assistant in Agronomy.
F. G. KRAuss, Superintendent ofExtension Work..
F. A. CLOWES, Superintendent Hawaii Substations.


i~ .II


1 Resignal October 27, 1914.

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IMwAn AcowaRns EXP=UNTmr &rAow

retho honor 0o submit here,"" *nd agggeicA or pub-

emboiestBenmposdn.40f Nitrogenous ,SubnAnmxw in
%MW' hada-,Moroxc'dordini heeded-
,Xejj icbio s

rn which plantit dra* -nitroe othigowh
sen~thio ls results 4f a stdy, 9!. the jercetages of/

saiecovfrmthed int6.thana wacmp s tetioon of aiuognc

Special Agent io hArwge.

Vjrector Sjaee Relatong Service,
U7 Department of Agriculture, Ifaehington, D. 0.

Publiation authorized.
*D. F. Housrow,


Series II-Anmoiifiacation it soil. ... :...... .. .::...
Series III-Ammbnification -of equal ouats of itrog &ti i
, Series IV--Am onificationin soil uxde.iar:,aeh otx4 t 4 I"
Series V-Ammonification with ,qual amoPz1t 1f "4 wyo:ue t:'::
nitrogenous materials................ ..... .. : .... -
Series VI-Ammonification with varying amu'ts of casel...
Series VII-Ammonification of casein duinig different length of. 1
Series VIII-Ammonification of caeein in silica sand........ i-.v.;.:i
Series IX-Ammonification and hydrolysis of casein....... ,.
Effects of bacterial action on different groups of nitrogen csompounnd .,.,.!i
Casein............................................ -.... ... ...
Dried blood.. ....... .. ........ .... ............ .. -....;,
Soy bean cake meal...--............................ .. .: --- ..
Cottonseed meal..--..------- .----.. --.. ....-.......--... --------
Linseed.meal --------------------------------------------------. 1

Globulin from cottonseed meal....-..-..-...... ;..
Zein from maize --- -- -- --- --
ein from ma ..--------------...... ...- ........................ ....

S. .......

=" ,**

...." .. .,:.
FIGURE 1. Diagram showing the ammonification of different amounts of ca-seis



... Il


.chagesproduead in the nitoogenous substances of
siteiaare Oof ret-mimporuance.' Asjaswell~ktowu,,the oAn
ago amonis attate ad niroen ut the snem
of wMchknge, which are probably. of graiprace
aprletyunderstood, 'It. appears thatO proitoins 'underg,
decomposition Mn soilw that which takes place
,digeto, and!U it s igl prohb~ab thatauoifcto
by hydroljrsis. From, the investigatiouB of Schreiner. et
mai -'eenshon tat ftl, aeitof prtein. and nucleoo
deaageprodu~cta -r widely d bitrbted -nsoils; and Lawns
n resenly, found from ammoniiation -expe'rimenta with
that int the, early stages, of the action. greater AMOUn~ts. of
wm*e obtained.-by.dsiln a 1 per -cent hydrochloric acid
*ith, mwutic soUe than with magnesia.,- They held that the
Aieds, decomposeA. soluble hydrolytic -products (wmino acids)
,'by the bacteria. .At a later: stage, when the amino. acids had
biy been more, completely decomposed, the yields -of ammonia
two methods' were, more -nearly equal.
aprobablitha niot alofheirgn in a given protein is
susceptible W: amonification 3 and that 'different proteins
decomposition at different rates, even when all other con-
are qua. n amoniicaionexperiments, for example,. it-
VA ldom' that more than from 50 to 60 per cent of the. nitrogen
added is recovered as, amoiad different nitrogenous substances
ieldamapnisat greatly different rates. As is "well known, the mi
Vorfion of the nitrogen. in soils occurs in organic forms and has pre-
opay existe as vegetable protein. After the organic matter of
1606 U been acted upon .by: bacteria for a time a; residue, rich in
if~gi, nd comol -aldhms left -which undrge further
mpoitin a avery slow rate. Moreover, the rate of formation
of am i from humtus a .ppears to depend to some extent: on the
66adiie undei* which the hdmus WWs itself formed. Generally acdd
wa ~~~. 0. DWP igt.Mr, sur. fta SOIL Z4, sot a SL
as EdMwa t stoLBM told.) 2. AhtV *7 (119U)#n KSA..A

in the tryptic digestion of edestin from: cdttion sieod l.
the tyrosin had been split off at the end of 2 Ays, wiW1
ceit of the glutaminic acid had been hydrolyredi. '
From previous work in this laboratory :it has .bneras
Hawaiian soils contain relatively greater '"amuduts of rsi
smaller amounts of basic (diamino acid) nitrogen than
proteins. From this it has been suggested that a study of...
changes produced under the action of bacteria might throwimia
light on both the availability of, and the nature of bauteNri.
on, nitrogenous fertilizers. Various factors, such as" ,1 a's
aeration, the acidity of the medium, the carbohydrates pr
synthesis of proteins in the body cells of the bacteria, the a .bsi
of organic nitrogen compounds in varying degrees by plants,
complicate the problem as to render very difficult an interp i t
of 'the chemistry of bacterial action in soils. It has; been i r
however, that by also studying the rates of decomposition'i
varying conditions some light might be thrown on this qu.

The materials used in this investigation contained the fo'
percentages of nitrogen: Casein (Eimer and Amend), 1?.40 pP,
dried blood, 13.29 per cent; soy bean cake meal, 8.28 per cezi
tonseed meal, 5.10 per cent; linseed meal, 5 per cent,..
In order to insure maximum aeration the first series of. tei.
conducted with the use of silica sand. One gram each of thej
nitrogenous materials and one gram of calcium carbonate were;.i
roughly mixed with 100-gram portions of sand in,tunblers. T.J
1 Ztschr. Physiol. Chem., 46 (1905), pp. 159-175. s Hawaii Sta. But 83 (191: :;:
.... : .N.ia
":....". :

.,* -:.:. ....**.* :.:. "' ..; ..Si' i' -*, .." .:

i iud
#,o 0 thi
Th1~0$e ,I I

041 t
vvlp swoAitetbtb t Op
VW.miia Q' pr
*Jpe1ossA oa u, n en~itr

ito o~ a
IDed 80_CCL Sm

clay soil trom wmicn tile inlusions were maae m u.psE
series. One-hundred-gram portions were eac thor
with 1 gram of the nitrogenous materials and 1 gram. f c.. .
bonate and placed in tumblers. Sterile water was adi dedI
bring the moisture up to 25 per cent. After incubati.ngt |.
the ammonia was determined as in the previous series. On.
day the nitrate was also determined in separate portions bml
disulphonic acid method, and was-added to the average
this period in estimating the total ammonification.

Amount of ammonia formed in soil from different substwaes.

Nitrogen as ammonia fzi:.:r:
Period of inu- Method of determination. Dr.e.
baton. Dried BoCye W#o
Casein. blood seed
S meal. e, i

Distilled directly with MgO....... 50.8 4.1 7.8 6
HCI solutions distilled with gO.. 381 2.2 5.6 4:8
2days.......... 40.0 4.2 8.1 &2-
s........... solutions distilled with NaOH {
4o.3 4.5 V.8 &.6,
Distilled directly with MgO....... 69.4 24.1 21.8 1L .
4days .HC solutions distilled with MgO.. 57.7 18.5 20.4 9.
HCI solutions distilled with NaOH { i
60.5 21.6 19.9 10. .
Distilled directly with O....... 68 6 42.4 29.4 10.4
HC6 solutions d= 'ith MgO.. 59.1 40.9 26. 3' 9
6 days......... 28.8 .
dys.......... HCi solutions distilled with NaOH { 66 .6 28.8 10.0'
56.8 35.2 23. 8' 9A8
ay ........ solutions distilled with MgO.. 50.1 b 8. 9 66 7.8
Sdays........... HCIsolutions distilled with NaOH 56.8 40.0 27.2 7.
9 days.......... Nitrate N................... 4.4 7.4 0 :0 .8
9 days .......... Total ammonfication....... 62.3 56. 4 .33.9 -1.8
Per cent of total N converted 50.2 42.4 40.9 27.1 '
into NHa.

It is not surprising that practically as much ammonia was obtained by .distiflgthte. i
magnesia as with caustic soda, since the acid amids are decomposed by each alie, aam4 "
protein cleavage products, cystin and'arginin, yield ammonia to caustic soda. Cystin s n .J
amounts in most proteins.
& Not used in average.

:. .: ''.. :]:iy.:*
1...:.,.: .:: ::.
j j. :: t.:.~

unhai recovered from tme different) mAtte"~l
Whe relative rates, of ammonifltatio'n throttghmot
Utms to, those of the, previous series. '' in the oasei_
blc, and, soy bean cake modl, lior which the Irgeat,
were obtained inthe sand cultures, still greater,
id in the soi. The cation Of cotton-,
Abdh~4 mod, was mmorenearly equal at e"c pLrod,
tesamo as that: which took place mi saxd. T~he
'were tongiderably, less than from the sanid,, due
Absorbing: -power of the clay A Omal mon
see bu itwasnot poortional to.amniia
'daa orthe total ainnmonification show, a wide difference,
Aaoop~siion of the materials;:50 2 per cent of the nitrogen
",Wa coverted. into ammonia,'42.4 per cent ME dried blood,
_entinsoy bean calke m eal, 27.1 'per cent in. cottonseed
26oceat i lined meal. Thus the availability of these
aued by, annoifcatin vais greatly.
ion, V&4. ,agesiaj,_specjally mi those instanes
Poxfiely Wag amunts of ammonia had accumulated,
bdraly more. DmonA than the distillation of hydro-
exrctse but', again, distillation. of the latter with mag-
yiede -smuch amnnonia as8 distillation with caustic, soda.
Wovd eem Itherefore, ,that direct distillation with magnesia
a tr ins~ure of a mmonification i~n clay soils than indirect
of hdochloi i-acd solutions. In all of the subsequent
d this bulletin the former method was used.
serio te nitrogenous materials were added so as to furnish
amont nitrogen (132.9 milligrams). These and I grain of
carboat were mixed with 100 grams of the same fresh soil.
brigig the moisture content to 25 per cent and incubating
beor the ys ofamna were as follows-.
Awont of ammonmiafrom equal amounts of niftogen.
[Averag of 2 samples.
Nitmogen as ammioniafom-

....... ............ ........... aA l ilt.9 173 .
..~~. .......... .. ..... 7 864 798 .
......... ... ..-............. 1 080 84.6 8
............... 74,12 AGL6 SB. 40.3. 4L*.
rWCm ftwXawre a .... L9 4. L7 30 X

milligrams) and 1 gram of calcium carbonate weree'mid
gram portions of soil as in the preceding series.
Amount of ammonia in soil under anaerobic conditions.
[Average of 2 samples.]

Nitrogen as a n"mwumiio ,,,"tmi
Period of incubation. Dried Boybean Ct
1. blood ake meal aedw
.072gm.p 1 gLm., m. 2.606 gpt

MgMr .g. My. y.: (f#
2 days............................................... 8.5 2.0 3.5 S8
days ............................................. 47.3 6.6 0.1 .I
6 days............................ .... ..... ... 62.4 13.7 14.2 .
9 days ............................ ...... .... 70.7 16.3 18.6 114 l!
Per cent of total N converted into N a....... 5.2 12.3 14.0

The above data show that under anaerobio conditions..
ammonification of casein did not begin until after two days' a
ing, but it then was approximately as rapid as under aerobic
tions. With dried blood, soy bean cake meal, cottonseed meal
linseed meal ammonification took place at greatly reduced
throughout the experimental period. The percentages of the:i
nitrogen converted into ammonia were as follows: Casein 53i.
cent, dried blood 12.3 per cent, soy bean cake meal 14 per'
cottonseed meal 8.5 per cent, and linseed meal 6.9 per
comparing these data with the preceding it will be seen that."x
conditions greatly retarded the formation of ammonia froEi
....... .-:* ... .

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li!UR t
A'Sw&o 4 1
"it O iet

11 19

"/i-.h miiosob~tWadtotm efjl
in eeWds `a'66qs
me 91I~ nqe~isix h
er aiiD'"th ai
ouis0c n 8 aiih n ga .cku~
fOid16r!'ot!U fteso&smi vptI
4(uu-W bia im te ae.oh
tUpoiu eis
**Mi oW4vaMdMU o owOwad "a"S

i/ei f app

Wmoe samna*m
tisonI re ko ~ m M2O
Paotv 4L m,
1W gm IW M. Si i I A~gn

thus eliminating a portion of the nitrogen present, Mis in it# ..
of their demand for nonnitrogenous matter.,
It should be borne in mind that the nonnitrogenous coti"'
of the above materials were made up of different chemical oe~i "
including fats and carbohydrates. The exact effect of. fatls
known, but different carbohydrates produce widely differed .
In general, soluble carbohydrates more markedly depress
cation than insoluble forms.3 The nature of nitrogenotus
ents in these materials-also differs considerably, and the. as
which they undergo hydrolysis, with the exception of casein ,.
been extensively studied. As will be shown later, the prodUi4
acid hydrolysis vary considerably. Since hydrolysis is
essential as preliminary to ammonification any differences.l.
rates of hydrolysis would probably be reflected in the rates of


In the preceding series the yields of ammonia from practice
same amounts of casein varied from 50.2 per cent to 56.9 .per .
the total nitrogen added. In the following series the caoce
-of casein was varied and the same amounts of soil and eaMia
bonate were used throughout. For.the purpose of re. ci.:.
1 New Jersey Stas. Rpt. 1911, pp. 193-212.
2 See J. G. Lipman et al., New Jersey Stas. Rpt. 1909, pp. 166-169.
a See Lipman et al., New Jersey Stas. Rpt. 1911, pp. 193-212.
SThe fact that peptone has frequently been found to ammonify more rapidly than di4qd li
seed meal may be due in part to its being a partially hydrolyzed substance.

; 0' : :.
.. "
A :;!::,i::

eRik^ a efr with the following resulted:
Asowof amWmonefb*aejkoa varying, qwhdtWte of masen.

Per cet Of pet Mant of
of qnga otlN Amount OfWN found as total M re-.
0100 asIoa r-09I

4mumals. eaveral as adL ammonia, Covered ws
0--1 il73l 24. O.
3 W.

4L4 is6.

LO M It. 517-0
S1, amiple Only.
g s 6f total: nitrogetn reovered as ammonia, creased
of casein inraed,, varying from 48.4 per cent with
0.2 gram, to L65.9 per
cent with 3 grams (see
fig. 1). Loss of am-
000 monia by evaporation
was not important,
since the percentage
yields, were greatest
where the greatest
concentration of am-L
monia occ urre d.
Since almost no nitri-
fication took place in
AS any instance, it seems
42 a#" A# 45 A0 2-6, 80
OeWs ar CoWSW wAMS reasonable to believe
L-Dh~v -howg the awmmnifeation dz eret amounts that as the amount
ofeCagan. **
of casein added is in-
a' demaing percentage of the total, nitrogen present would
a onsmedby the bacteria and consequently higher percentage
Ofm he obtained. 4 The yield from I gram (57 per cent)
,4gre clodywith that recovered, in preceding- seres,
Ili Iirit as begnm at the 9&me time as Serie VI, using the same
Onepamch of cwsemin, cacium carbonate, andmgniu
-m~su.wasmixed with loo-gram portions of soil, sterile water

Practically, the same amounts of ammonia were fti
given amount of casein in four days as in longer periods
from 1 gram were 57.3 per cent in four days, 59.8 per cent
days, and 58.6 per cent in twelve days. By adding a i
on the fourth day and allowing the action to continue ior
longer 60.8 per cent of the total nitrogen was converted intofa.i
The portions treated in the same way but allowed to stand' i-gi
longer gave 57.1 per cent yield of ammonia. Finally when ~1
was added at the beginning and after four and eight days,repei
61.8 per cent of the total nitrogen was converted into a o"ni.a;.
The above data show, therefore, in common with the.pi
series, that increasing percentages of the total nitrogen were conv
into ammonia when increasing amounts of casein up to 3 grain
acted upon, but whether this fact was due to partial suppress
the nonammonifying organisms can not be positively stated.
is evidence, however, that under the conditions of these exp
the organisms feed on the organic nitrogen of casein rather thaai
ammonia after it has been formed.
The amounts of ammonia recovered from casein in the p
experiments usually did not exceed 60 per cent of the nitrogen
and reached a maximum point by the fourth day. In order to
further light on this subject a series of experiments was carrid
with silica sand, .provision being made for absorbing
ammonia was volatilized. The decomposition took place in
stoppered bottles through which a slow current of air was dr i
means of a suction pump. The current of air was first drawn.
sulphuric acid to remove traces of ammonia and after passing
*"":* "


with "te cet ameeio sid, ffatering, hiwd ,thwovogWl
eithaloholaidOther. After dryiag in vtouumhiveviy
4thw prodwes was found to contain oilyJ 18.50 -pmr 6=t
beeigetat onsdexiable nimpuri ties- still remained&* (Poise
UA20 per "ant N.) .v
-4f thel "ame was mixed with 500 grams of. ilics sind
mnoistbre brought about by adding 50 eubic centimeters
haionprepared as in Series 1. After incubsting at 280- 0.
the contents of the bottles were acidified with 1 per cent
,thoroughly shaken, filtered, and washed.. 71we residue
*wtracte with tenth-normnal sodium hydroxid and the
tosacidified with I per cent so'etie cd opecptt
i he solutions were combined with those above. Am-
etdetermined i' the, solutions, sand residues, and the
said by distilln with an excess-of magnesia.
VV *owalamo of ammonia frmedfrom casein in sIniea sand..

X XsN~ as NHe H as NEs TotalPe nto
---- inte voai-to r t

.M.. Mg. Ia.

------ ------------------- 271.6 0N6.6 J16-2L6 M4030(L8 63.8
........... ................ 273. 96.6 M64 4a6. M6l
...... ----------- 32&98 55.7 43.4 423.9 62.8

digtlyhig~i-yield of ammonia was obtained than previously?
being 64.2 per cent of the total nitrogen. Since no casein
be precipitated from sodium after bacterial
,the conclusion that the entire amount of casein added had
uleronehydrolyiis is justified. Tno nature of the undetermined
anerMains' to be determined.


+ Te purpose of this series was to study the relations between'the
Of amonfication and hydrolysis of casein. A stock solution
-,saoin purified as in the previous series was prprd by dissolving
) grme i 150 cubic centimeters of tenth-normal sodium hydroxid,
an iluingto 21100 cubic centimeters. One hundred cubic centi-
ptrportions were placed in 300 cubic centinmeter Ere eyer flasks
10 cubic centimeter of soil infusion added. -.After .h&iath
h weare loomly stoppered with cotton plugs and Mincuhted at
n 1C 'm( fhli!ssws eaue rpecap igt

[Average of 2 samples.]

Period of Incubation. N present. prsP. l
---------------_---- :;,, "*". ..:|

Hour .................................................. 94.3 % .4 -
3 days................................................. 943 89.6 :7
9 days..................................... ... 94. 3 87.6 &":A B
----------------------- -..!!6i--

The above data show that in one hour's time 4.13 per ceAt
nitrogen underwent autohydrolysis, and this was increased :
standing for 9 days to 6.79 per cent.
The effects of bacterial action on the ammonification and hi.
ysis of casein are shown as follows: !

Some results of bacterial actio n ocasein.
[Average of 2 samples.]

N found pJPer.oent juT
Period of incubation. N added. N fo N preci- N o
as *NH. ated. ed.

.Mg. Mg. Mg.
Iday...................- ..-................ 94.3 0.0 90.A I.
3days.....-----------..............------------..--.---- 94.3 .6 9L1 G.
days ............................. ....... 94.3 2.5 75.9 2.05
5days...........-.........-....- ......... 94.3 9.7 29.6 104
7 days..-........... -........ .... ......... 94..3 5 .1 13. 8 S
9days.................................... 94.3 58.7 17.6 2.
IdYS-----------------------------------9435. 76 8 ..5

Active ammonification set in after the fourth' day and te.
practical maximum on the seventh day, when 59.53 per c ient
nitrogen had been converted into ammonia. Active hydroly.t
in after the third day_ and was completed by the seventh d-i. r
this time the solutions had become quite opalescent, due to
abundance of cells of bacteria and fungi, and no precipite

17our. Biol. Chem., 11(1912), pp. 267-305.


All -- A

foreging- it 'is appaznt, that bacterial hydrolysis of
esamnonifieation and that the. former takes place eon.
erapidl-v than the latter..

experin.metg maxmumT ajmmonification usually
-Afroln four to-six days. Of the substances'tested casein
moetcmpletely Iaxnmoxiified, but as, stated above. usually
60per eto h nitrogen was converted into amna
experiments were made for the purpose of studying
'' of bacterial, action on the different groups of organic nitro-
d Two grain portions pf- the substances were mixed
grams, of- silica sand, soil infusionfs added -and incubated for
pirios. Ten he- sand mixtures were trainsferred-to 1,000
,cenxtimeter Ijeldahl flasks,'400 cubic- centimeters of hydro-
uagi added, and the whole 'boiled under reflux condensers for
ears.After filtering and washing the residue with hot water, the
were dihitted to 1,000 cubic centimeters and aliquots used in
termination: of :the amid, basic, an obsic, groups of ntoe
,employing the game methods- as were used in previous
on the organic nitrogen of Hawaiian soils.'
ever cv, the residues, left after filtration were patclyfe
AitingnAfh~w~ing that All the nitrogen present went into solu-
butA a Smaller amount was generally found than occurred in the
materials. Th-1is was probably due to the loss of ammonia
1volatilization during the course of bacterial action, and 'to the
position of nitrates, and therefore, will be considered as having
converted into ammonia. It is-possible, however., that some
,_ADcaition also took place.- The length of time that the different
were exposed to bacterial action varied, the purpose being
aalwi decomposition. to continue no longer than was necessary to
vigorous a onfication., Ammonia was determined in
portions. by direct distillation with magnesia. The original
trialsias were also subjected to acid hydrolysis and the group deter.
phistinsmade: as -above. All determinations were made in

The materils studied include casein, dried blood, soy bean cake
,cottonseed meal, and linseed meal whome nitrogen contents.
Ra i&Il O,

AmmniaN.......................... ..........
Amid N ....................... 1.4
BeAic N.............................. 2.12
N aonbas N...................-... 8.7.

.3-- .....3.. .
o0. 95 17.111 77
.6.381 .7,9 ..0. ,.S

The casein used was not gure. In addition to coii
soluble matter it probably contained small amounts f~kaa :
bodies other than casein. Pure casein from cow's 8 iWb.V
10.3 per cent of the nitrogen in the.form of anmids and:22.
as basic nitrogen compounds, while the above material y.i l
per cent as amids and only 17.11 per cent as basic compouwjS
The bacterial action was allowed to continue for three daysi
which time 31.96 per cent of the total nitrogen was concert#
ammonia. The amid nitrogen was reduced from 12.43 peiS
8.88 per cent of the total, basic nitrogen from 17.11 per centt
per cent, and nonbasic nitrogen from 71.59 per cent to 50.93-pa
Expressed in percentages of decrease we find that 28.57 per eeant
amid nitrogen, 55.19 per cent of the basic, and 28.86 per cenato
nonbasic nitrogen were ammonified. The organic nitrogen. :
ing at the close of the experiment was composed of 13.16i
amid, 11.34 per cent basic, and 75.48 per cent nonbasic
compounds. Comparing these percentages with the comp
the original casein it will be seen that the basic nitrogen coc
were decomposed more rapidly than the amids or nonbasic

Nitrogen content of dried blood and its bacterial decomposition mprodtw. '

Ammonia N..... ...............
Amid N....... ...............
Baste N..............................
Noubasio N..........................

Per cent of original



Per cent of total N.

- I







1 I

Per cent of
groups d4l i
composed. I'
.. ...

...... ..



- I -- -- -- 4 I


ibie table shows that 49.52 per cent of the total nitrogen
id Te amid nitrogen decreased from 14.97 per cent
Sit of Wte total, the basic diamino acids from 9.18 per
is per cent, and the nonbasic nitrogen from 75.84 per cent
S pew cent. Expressed in percentages of decomposition it is
l-6i 430 per cent of the amid nitrogen, 67.10 per cent of the
ittgfn, a d 4825 per cent of the nonbasic nitrogen wea
iued The organic nitrogen remaining was composedof
lpent ami, 5.74 per nt basic, and 77.75 per cent nonbaic
SapoA.nds, as compared with 14,97 per cent amid, 9-18
a4i a 7.8-4 per cent nonbasic nitrogen in the original

--.. .- .. ,; .1 .... .....
; i i "T .

AX 11 W.. j i:" .i A
ii, ~t i

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Ammonia N....,.................. ......... .. f l .,:-, ..
Amid N ........................... .0.78 0.698 1U 1aLit
Basic N............................... .. .39 ,4 8 Si
Nonbasic N..........- -.... .... 3.36 2.74 6. 8 F .
,.- :; i, '
The above data show that 28.43 per cent of the total '
ammonified. The-amid nitrogen decreased from 15.29. perF i
11.57 per cent of the total, the basic nitrogen from 18.83 per
6.27 per cent, and the nonbasic nitrogen from 65:68 p !:.t
53.72 per cent. Expressed in percentages of decompositdiS
found that 24.37 per cent of the amid nitrogen, 66.67 per
basic nitrogen, and 15.48 per cent of the nonbasic n
ammonified. Thus it is found that the basic diamino aai
ammonified more rapidly than the other groups. '


Linseed meal was subjected to bacterial action for seveI j
with the following results:

Nitrogen content of linseed meal and its bacterial decomposition products.

I Per cent o nal Per cent of total N.
material. Per cent of
groups d4
compoagd. ^H
Before. After. Before. After.
Ammon-a------------------------- ---1.99 39.80...........4
Ammonia N.................- .......... -1.99 :.......... 390.80 .....
Amid N.....- ............- ........ 0.83 .64 16.60 .' 18 0 22.89
Basie N...................... ... .. .62 .38 12.40 7.60 37.0 ::
Nonbasic N..................... ..... 3.55 1.99 71.00 39.80 43.6

It will be seen from the above table that 39.80 per cent
nitrogen was ammonified. The percentages of decompositiofi
that 22.89 per cent of the amid nitrogen, 37.09 per cent of tEl
nitrogen, and 43.66 per cent of the nonbasic nitrogen were ammo
The organic nitrogen remaining was composed of 21.26 per
amid, 12.62 per cent basic, and 66.11 per cent nonbasic compo
as compared with 16.60 per cent, 12.40 per cent, and 71 per t:en
respectively, in the original material. Thus it is shown, in contra

verded sped orerapdlythan the iroe

nn tion experimets mith this material indi.
nh itrogen constituenta would. be. decomposed mom,
mn the mxteriala reported above. After ineubsti'ng
t'icll no ammonia was found. Consequently h
onW" avowed to tale place for 12 days,. but even then,
a6nont of amnmo-nis was formed.
'111 dof woomit meal ad itr bacterial decmnposiain progduts.

Maerial. Per Vent of towa N.: Per cent Of Per Cent of
groups da .ogni
Befoe. Afte. Bfore Afercomposed, rWa action.

-------------------....... ...4 ..... 0 2 7.27 ........... .. ....... ....
.t .. ........... R 8 .37' 12.00
---. --.- -2 .41 15.70 12.43 21.15 13.39
.,.......s... ....-... 2140 2.29 72-7Z 6W.09 5.00 Il.51,

ilool seem that thef carbohydrates and. fats protected the
boisfro~m' bacterial decomposition, since 641y 7.27 per
the toal itrgenwas found as ammonia,i and the absolute
4f nitikogen iA, the different groups were only slightly-
;,frean* those in, the original material. But the magnitude
4dp"rmen~t4&l eirtor 'was relatively too great, to justify YPositive
data shw, howeer, that a higher percentage *of the moi
defived from their basic nitrogen group than in any of their previous


Tke globulin was prepared from cottonseed meal by extraction
Wb 10 per cent solution of sodium chlorid, then precipitated by
sajtaitiug the. solution -with moium sulphate, redissolved in
*odiunb_ bborid solution, and dialyzed. The product was washed
*ith alcohol and other, and dried in vacuum over sulphuric acid;
U~wA oiU. sspriy h irgncotn.Amnr
oo~inudfrti as

JVZLLLU. tVLlV IUIUJ v LLULcmU T, C U VL6 OLUIi W't L5U io.flf W L U Vaww..i:
for .24 hours for complete hydrolysis.
The above data show that 27.77 per cent of the i..
converted into ammonia, and that 42.22 per cent of the
per cent of the basic, and 17.48 per cent of the nonbW
compounds were decomposed. The organic nitrogen.
after bacterial action was composed of 8.81 per centi
per cent basic, and 75.40 per cent nonbasic compounds.
the above data with that obtained with the use of cottoasei
it is of special interest to note that globulin, when sepa
the other nitrogen and nonnitrogenous constituents .of ctO
meal, undergoes bacterial decomposition in very much.,i
way, as do the nitrogen compounds of cottonseed meal as 4
In each instance the basic diamino acids were decompo
rapidly than the other groups. The amids, however, wee .
posed more rapidly in the globulin than in cottonseed. meal'.
With the exception of linseed meal the basic diamino aceid: d
preceding materials were decomposed more rapidly than th'aio
monamino acids. The basic nitrogen in these materials tarie4
9.18 per cent to 23.02 per cent of the total nitrogen;. In ordeif Mt
the decomposition of a substance containing still less diamiho n
zein was prepared from maize by alcoholic extraction. The -pe
was not highly purified but the analysis shows that practc
the nitrogen was in the form of zein.

't till

4 ~ ~ ~ ~ ~ ~ ~ ~~~Oal AV nysalamie faaonawr od
per i
tleq, hcp

1l rol 4

A x

A kofa&' The data show that, the basic diamine- aid
to asliht eten;-but in this, OAsete nd
n tkedly dbbbnos ed. M6' &ereseis in. ampid nfb
itdearmay not hivobeo oen eniey de t mo
Ampe any, mid compounds that were split off b14 64 $c
ht nh~kite, oud hav ben debpsd -.and -deter-
6hibiv 4deng wiih that: dlfctrdiaYfre by the 'bacteria.
Aboea large portion of the nitrogen in the materials
th inodfid 0+ At east -did, not occur at any one time as
Aiield of ammo. mcasein in e -
boi mierillyinceasd y prolonging the fime of doecmi-
9%yolidf6r days. -But the m-ssation of ammonificatfion lvas
4 te ccmuato ofpionou by-products c, sic he
and third gram, added after the ammonnification of one gramm
*t"B gof er each ammonified to a slightly greater extent
'fistgra..Itseems P:robable, therefore, that a part of the6
6 ntroenin hematerials used is more resistant to ammoni-
a tAn oters It sghoulld ailso b6 remembered that putrefac-
dewom-position usually takes place to some extent in the ordinary
XUMtion experimetnt,) which probably ipesults in the formation
Inamatie protein claage produ( tyrosin, phebnylalanin, and,
HAne at first;, labor these are 'decomposed Minto indol and
WI&O, either 'than lieig aimmediaty converi~tedi :into. ammona It
-*bo that -a porton: of the nitrogen was assimilated by the
presnt'but whether the assimilationi of ammona o
lorms took -place 'cM not be definitely 'stated. The after
1&6 mxnore proibable In eter ase it i's reasonably. er tamnta
-40611+l organie, hitrogeni.

enOgW 7EE~b 1S-

Abe i;iyiss oorpdytai.f

* WU 3LjrjVLL tUJ. OjLLLLfLIJflIj LJL J LULLJ. UJI1. LULL IJA J 'J'al Caj. wJ.qfCMI s~iaija
meal. During the nine days of .the experiment 56.9
nitrogen in casein, 49.3 per cent in dried blood, 48.7 .
bean cake meal, 32 per cent in cottonseed meal, and 34,G
in linseed meal were ammonified.
(4) Under anaerobic conditions all of the materials Wrl
fled very slowly during the first two days. Later the; as i
averted into ammonia approximately to the same ete:iii,
aerobic conditions, but the other materials were decoinm p
less vigorously.
(5) With equal amounts of both nitrogen and naoni
matter present the final yields of ammonia from the: dif
trials, with the exception of dried blood, agreed closely, but.h
decomposition of casein was still much more active thqa":
substances. The yield of ammonia from casein on the: n
only 31.4 per cent as compared with 56.9 per cent in the
starch, and the ammonification of dried blood was reduced
per cent to 18.9 per cent. It has been suggested that their
fying organisms are able to utilize carbohydrates tip some
sources of energy. If so, smaller amounts of anmonp. woul
quently be split off from proteins in the presen'pe of _ic
Hence the carbon-nitrogen ratio would materially. a et
formation of ammonia in soils.

.:4: ..

tr s day s when mixed with soil (see Series IV).
STiiiX deter ln tion of the different groups of nitrogen com-
Iefore adnd after bacterial action in casein, dried blood, soy
a ecke meal, cottonseed meal, linseed meal, coconut meal, globulin
a cottonseed meal and zein from maize shows that, with the
option of linseed meal and zein, the basic diamino acid nitrogen
rteed into ammonia more rapidly than the nitrogen of other
i.p.: With casein, soy bean cake meal, and cottonseed meal the
S.trpi ammonification of the basic nitrogen was especially
liable. When' this fact and the above are considered in connec-
"O a comparison of the organic nitrogen of soils and vegetable
ine, it becomes apparent that all portions of the organic nitrogen
bd i:een t materials used as fertilizers and green manures are not
i seeptible to ammonification. It is evident, therefore, that
iu fators inherent in the nitrogen compounds themselves
e the availability to some degree. Further investigation,
.a study of the decomposition of individual amino acids
iamids, is being made.

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