Patented Dec. 20, 1932
' 1,891,750
UNITED STATES PATENTOFFICE"
‘FRANKLIN M. CORNELL, OF BROOKLYNQNEW YORK, ASSIEGNOR '1‘0 BTW LL'BO
BA'I'OBIES, INC" 01'‘ LONG ISLAND-CITY, NEW YORK, A. CORPORATION OF NEW YORK
BACTERIAL PRODUCT
_ Il'o Drawing.
Application am June 1c, 1928. Serial No. 286,061.. ‘ '
This invention relates to bacterial products
useful in the inoculation of seeds or soils
with bacterial, particularly Bacterium radi
cz'cola, in such manner that the bacteria will
continue to vegetate and grow in con]unct1on
with the plants resulting from said seeds or
stant h drogen ion concentration of the prod
uct. Ilumus contains substances‘ apparently
actin as nutrients and whose chemical na
ture is not well understood. The variation
in chemical composition from one ‘sample of
humus to another is too great to permit the
growing on said soil.
maintenance of a predetermined standard of
It has been the practice heretofore to pro-, quality. Humus cultures as heretofore pro
. duce bacterial cultures for soil or seed in duced are therefore inde?nite and inconstant
10
oculation by treating humus or the like with in chemical composition and bacteriological
the desired bacterial culture in liquld or Jelly‘ action, cannot be sterilized to advantage and
form, adding substances considered bene?cial
to the growth of the bacteria, adding suffi
are inconvenient to use.
the product to what was considered as the
as follows:
.
The product of this invention consists in
cient water to bring the moisture content of general of ten types or classes of ingredients
15
" _ proper percentage for the continued growth
of the organism and mixing. thoroughl, .
The product is used by ?rst wetting the see s
-
1. An inert base.
2. A coloring material.
'
3. A bacterial culture produced in any suit
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either with water or a dilute solution of sugar able manner and containing the particular
and then mixing with the bacterial product strain or variety or strains or varieties of the
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so thatthe product adheres to the seeds car
rying the bacteria with it. It is then neces
sary to dry the seeds before planting.
The bacterial product or humus culture
as made and used at present has the following
disadvantages:
The use of humus as a car
micro organism that is to grow in conjunction
with the above mentioned plants.
'
4. A nutritive substance or substances for
the purpose of keeping the bacteria alive over
a period of time.
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75
5. A substance or substances stimulative to
rier introduces a great number and variety the growth of said bacteria.
of undesired micro organisms into the prod
6. A substance or substances promoting o
uct which micro organisms oftentimes re tending to promote the growth, multiplica
tard or entirely prevent the growth of the tion,' and functioning of the bacteria under
desired bacteria. For this reason these un adverse circumstances as in the presence of
desired micro organisms are referred to as compounds of nitrogen.
contaminating bacteria. The humus culture
7. A “buffer” substance or substances for
causes contaminating bacteria and fungi to the purpose of controlling the reaction or
continue to grow. Humus cultures as pro hydrogen ion concentration of the bacterial
.duced above do not adhere to seeds without
additional treatment of the seeds or of the
8. A substance or substances for the pur
culture. Sterilization of humus before use pose of causing the bacterial product to ad
in‘ manufacturing a humus culture results here to' seeds without treatment with any ad
7 product. -
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in many cases in a chemical alteration of the
humus in such manner that it causes contami
85
>
ditional substance.
'
9. A substance or substances for the pur
45
nating organisms to grow more rapidly than pose of preventing or inhibiting the growth
otherwise. Moistening of seeds prior to in of contaminating micro organisms.
oculation necessitates in many cases the dry
10. A substance or substances permitting
ing of the seeds before planting with conse said bacteria to retain such characteristics
quent loss of time. Chemical action of add and‘ functions as they possess when growing
ed substances upon the complex mixture in the soil or on plants growing in the soil.
knownas humus may result in the forma
The inert base or carrier consists essential
50
bacteriological action, resulting in incon asubstances of simple chemical composition,
tion of. substances of unknown chemical or 1y of a ?nely‘divided substance or mixture of ‘
95
a
f2,
1,891,750
substantially sterile as produced or procured,
notchemically' altered byexposure to tem-'
peratures customarily used in sterilization,
having no determinable effect by virtue. of its
chemical composition upon the micro organ
isms supported thereby and having no in?u
- ence or a determinable in?uence upon'the hy
organisms to a lesser extentthan to the de
sired bacteria.
_
The substance or mixture of substances
stimulative to’the growth of the desired bac
teria'is mainly for the purpose of reducing
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the amount of bacterial culture ordinarily
introduced into seed and soilinoculants. It
drogen ion concentration of the bacterial must be of such nature and in such amount
as will permit the bacteria to remain alive,
sarily dark in color, of porous or absorbent functioning normally over a period of not
nature, of such physical properties as will less than six months. It is preferably of
cause it to adhere to seeds and consisting of__ such chemical composition that it will not
or contammg'a'compound of silicon. The react chemically with any other constituent
principal purpose of the inert base is to act of the ‘bacterial‘product. It is preferably
product. It is preferably, but not neces
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75
is as a carrier ofthe bacteria. Sand as ordi of such nature that it will not stimulate the
' narily found is not sufficiently '?ne- for use
as a base.
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growth and functioning of contaminating
‘micro organisms to the same or greater de
The coloring material consists essentially gree than it stimulates the growth and func
tioning of the desired bacteria.
‘The formation of nodules containing
ture of substances ina ?ne state of division,
of simple chemical composition, substantial BGGZ'PEPZUWL radicico'la upon the roots of
ly ‘sterile as- produced or procured, having no leguminous and other plants has been found
determinable effect by virtue of its chemical to be greatly inhibited by the presences of
composition upon the micro organisms in, compounds of nitrogen. A substance - or
the ?nished product, not chemically altered mixture of substances is therefore introduced
by exposure to temperatures ordinarily used for-the purpose of promoting nodule forma
in sterilization and having no in?uence or a tion in the presence of nitrogen compounds.
of a black or dark colored substance or mix?
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determinable in?uence upon the hydrogen' The substance or mixtureof substances must
ion concentration of the resulting product. be of such nature that it is not injurious to
It is preferably but'not necessarily -of po the bacteria in the bacterial product or to the
rous or absorbent nature.
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It is not necessa
contaminating bacteria. In order to pro
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90
95
plants whose seeds are inoculated therewith. '
rily of such physical properties as will cause
The substance or mixture of substances for
it to adhere to seeds. Its main function’ is the purpose of controlling the reaction or
to act as a coloring matter when the product hydrogen ion concentration ‘of the bacterial
is applied to seeds so that one can diiferen- ‘ product must be of such nature and in such
tiate between inoculated and uninoculated amount that it will not be injurious to the
desired bacteria nor to the seeds or plants to
seeds.’ The product of this invention is
bacterial product containing a minimum of ,
85
100
which the bacterial product is applied. It
is preferably of such nature-as will keep the
duce on a commercial scale ‘a product having reaction or hydrogen ion concentration of 105
this highly desired characteristic it has been the bacterial product either at the optimum
found of importance to use as a base and as ' value for the growth of the desired bacter'a
a coloring material substances which may .be or at such value as will permit the growth
procured or producedin a substantially ster of the desired bacteria while inhibiting or
ile condition. Thus‘ eliminating to a large preventing growth of any or all contaminat 110
‘
'
extent the possibility of incorporating in the ing micro organisms.
The substance or mixture of substances for '
product an undesired amount of contaminat
the purpose of causing the bacterial product
mg bacteria.
’
,
As the bacterial culture may be produced to adhere to seeds must be of ‘such nature
50. in any-suitable manner the method ofpro
as to have no injurious action upon the 115
’ ducing it does not constitute a feature of this bacteria in the bacterial product or upon the
plants or seeds to which the bacterial product
invention.
- ‘
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The nutritive substance or mixture of is applied. It may be a liquid or powder
' nutritive substances is of such nature as will and is preferably of simple chemical com
position, not chemically altered by tempera
retain the functions normally possessed by tures customarily used'in sterilization and
55 cause the bacteria to grow, multiply and -
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the bacteria when growing in conjunction having no in?uence or a determinable in?u
with the plant whose seeds were inoculated ence upon‘ the hydrogen ion concentration of
therewith. It is of such nature that it will the resulting product. ‘It is'preferably pres
60 not react'chemically with the ‘inert base or ent in such amount‘ as will cause the entire
the coloring material used. It is mainly‘ for mixture used in the bacterial product to
the purpose of keeping the bacteria alive over adhere to the seeds being inoculated With
a period of time of not less than six months. out treatment- of the seeds or of the bacterial
’
. It islpreferably 'of‘suc'h nature that it will product with any added substance.
65‘ act as a nutrient for contaminating micro
‘The substance or mixture of substances
130
3
1,891,750
permitting ‘the desired bacteriafto retainv
Any or all of the ingredients as obtained
may be tested by appropriate bacteriological,
' such characteristics and functions as they
possess when growing in the soil or on plants chemical, or physical tests to determine their
' growing in the soil is preferably ofa siliceous suitability‘ for use in the bacterial product.
70
5 nature. Its main purpose. is they retention Any or. all of the ingredients with the single
of virulence-by the bacteria.
I exception of the bacterial culture may be
_
One instance of substances which may be brought to sterile conditions at any time prior
used for the product is:
'
to additionof the bacterial culture.
The above mentioned ingredients may be
lo Inert base-Talc
mixed or ground together in any suitable
Coloring material—Manganese’ dioxide
* manner and added to the particular batch of
Nutritive substance—Maltose
Stimulant-Calcium carbonate
bacterial product being manufactured, in any
I
satisfactory manner, and in suitable propor
Substance improving resistance to nitrogen
15
Ammonium phosphate
tion, the method of manufacture/described
.
The inert base, the coloring material, the
>
“Bu?’er”—-substance for’ controlling. hydro- .below being preferred.
gen ion concentration-~Borax ‘
> substance or mixture of substances for caus
Substance to adhere to seeds-_——Talc
ing the product to adhere to seeds and the
substance or substances for the'purpose of
growth causing the bacteria to retain their normal
functions, are added in powdered form to a
and function as in soil—-Talc ' 'g
~
‘
suitable mixing machine and are mixed thor
These ingredients may be used in substan "oughl'y. To‘ this mixture are added the
tially the following proportions:
nutritive substance or substances, the sub
Substance to prevent growth of contamina
tions—-Copper sulphate
.
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Substance_ to cause bacteria to retain
2'5
stance or substances stimulative to the growth ' '00
Parts by
.
-
weight
Talc ______ __' ______________ _a_-_._' ____ __
9
‘ Manganese dioxide; _____ _1-__‘________ __ 10
'
_
Maltose __‘ _____ __~___' ________ __‘_~__.._-..__
1
so Calcium carbonate_‘-_; _______________ __
5
.of the desired-bacteria, the substance or sub
stances promoting'the growth of the bacteria
under adverse circumstances as in the pres
ence of nitrogen compounds, the “buffer” sub
stance‘ or substances for controlling the hy
drog'en ion concentration of the product and
Ammonium phosphate_v ______ "a ____ __,_ ‘ 1
Borax
____ __'-.__ _______ ____.___‘____‘>____ __ ‘ 1v
the substance or substances for the purpose
Copper sulphate___i_-~_'_i______,____i_;__ /2~ .of
preventing or inhibiting the growth of
a5. Moisture—Appr_oximately 25% of the total contaminating micro orgamsms.
< .All of the above substances are preferably 7100
\’ in the form of more or-less dry powders. The
, weight of the above in vredients.
Bacterial culture-Sufficient quantity.
entire mass of ingredients thus far-ladded'is
su?i'ciently mixed and to the mixture are
added su?icient bacterial culture and‘ water
Another and preferred instance of sub
stances which may be employed for the prod
4ouctisz'
'
'
.
.in proper amount to bring the moisture con~ '*
Inert base—'-Kieselguh1‘ or infusorial earth
Coloring material~.—Carbon black
‘tent of the bacterial product? to what is
deemed to be a' satisfactory percentage._'
vWhat is a satisfactory percentage for any
'
Nutritive substance-Starch
Resistant-to nitrogen-Potassium" alum
Stimulants'—Starch, ‘calcium chloride
batch of the product is determined ‘by-the
product characteristics desiredas the water '.110
“Buffer” substance—Sodium bicarbonate '
content exerts a marked in?uence on the . .,
virulence and duration‘ of life of the desired‘
bacteria.‘ Usually the water content of the
tions-Sodium bicarbonate ‘
v product is around 25 percent.
The mixing
0 Substance to cause bacteria to retain func is continued until the ingredients of the bac-'
‘- tions as' in s0il—-'—Kieselguhr >
terial product are thoroughly mixed and the
- Substance to adhere to seeds——Kieselguhr _
' Substance to prevent growth of contamina
1-15
These ingredients maybe used in substan product assumes a more or less homogeneous
tially the following proportions:
55
.
.v
i
'
d‘
'-
'
_ appearance.
,Partsby
7
weight
Kieselguhr _______ _'_____'_ ________ _._-____ 85
Carbon black____' _____ _..' ____ __‘__.-_____ 15
Starch 1
‘
_.._
8
Calcium chloride"? _________ -1 ______ --‘-’
'-
1
Potassium alum_..__'__' _____ _,__.;_____V____ 1/2
Sodium bicarbonate ______ _-______‘_______
;Moisture—Approximatelyj 25% of the total >
.weight of the above in
ients.
65 Bacterial culture—Su?icient quantity.
l
_.
While certain ‘preferred embodiments ‘of
.
120
the.‘ invention have been described it will be -
understood that it may be embodied in other
forms, thatvsubstances other’ than those spe
ci?cally mentioned may be used and changes
may be made in the, relative proportions of 125
the ingredients without departing, from the ‘
invention as de?ned in the appended ‘claims.
WhatIclaim is:
,
~
v
1. AI bacterial product containing ‘Bac
teriu'm radicicola, kieselguhr, carbon black,
130
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1,891,750:
starch, calcium chloride, potassium alum and
sodium bicarbonate.
2. A bacterial product containing Bac
terium radz'cicola, kieselguhr substantially 85
parts by weight, carbon black substantially
15 parts by weight, a starch substantially 8
parts by weight, calcium chloride substan—
10
tially l'part by weight, potassium alum sub
stantially one-half of 1 part by. weight,
sodium bicarbonate substantially _1 part by
weight and moisture constituting approxi
mately 25 percent of the total weight of the
. above named ingredients.
3. A bacterial product in powder form
containing Bacterium 'radz'cicola, an inert
silicious base, ‘a carbohydrate bacteria food,
said product also containing calcium chlo
ride to stimulate the growth of the bacteria,
potassium alum to promote the growth and
functioning of the bacteria vunder adverse
circumstances and sodium bicarbonate for
controlling the hydrogen ion concentration‘
.of the bacterial product.
' In testimony whereof I a?ix my signature.
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: G0
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FRANKLIN M. CORNELL.