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Katherine Golden Bitting.

The effect of certain agents on the development of some moulds online

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25



Histological Features



Agents



Kind



Penicillium
Alcohol



Benzoic acid

Sodium benzoate .



Boric acid .
Creosote. .



Formaldehyde . . .

Formic acid

Formic acid

Sodium formate. .

Sodium formate. .
Saccharin



Salicylic acid.



Sodium salicylate.

Alternaria
Alcohol



Benzoic acid

Sodium benzoate .
Boric acid



Borax . . .
Creosote .



Per
cent



Size of

germ.

conidia

in M



Formaldehyde .



5.0
0.1
0.2

0.2
0.04

0.1

0.2
0.5
0.2

0.5
0.2
0.2
0.2
5.0



0.02
0.03
0.2

0.5
0.05



0.1



Characteristics of development



15.2

49.4

15.2
38.0



15.2

19.0

9.5



7.6



14.0



11.4
41.8

13.3



13.6

13.3

15.2

9,5



Hyphae swoHen, distorted, walls tough; protoplasm
clear.

Hyphae larger than in benzoate, more easily broken,
distorted.

Hyphae and conidia swollen and distorted, no uni-
formity in formation of septa, some hyphae have
few, others many; protoplasm coarsely granular,
filling tubes; walls break readily.

Hyphae swollen, short thick side branches, blunt
ends, protoplasm finely or coarsely granular.

Hyphae nearly as wide as germinated spore, many
septa; protoplasm homogeneous, many fine vacuoles,
many short side branches develop into fruiting
heads, having few branches.

Hyphae attenuated, septa distinct; protoplasm ap-
pears solid, shrunken from walls. Fruiting heads
have stringy branches, conidia have refractive
greenish content, shrunken.

Hyphae swollen, coarsely granular, lilunt tips, many
broken, or, normal size, finely granular, many
vacuoles.

Hyphae swollen, coarsely granular, yellow, dis-
torted, badly disorganized, break easily. Nearly
all germinated conidia broken.

Hyphae swollen, short side branches, blunt tips;
protoplasm coarsely granular, disorganized; or,
hyphae normal size, protoplasm finely granular;
some cells empty.

Hyphae swollen, short undeveloped side branches,
blunt tips; protoplasm coarsely granular, disor-
ganized, walls break easily.

Hyphae enlarged, some much swollen slight dis-
tortion; protoplasm clear, homogeneous; conidio-
phores thick, stunted.

Hyphae and conidia swollen, some of the conidia
much elongated, few septa, tips blunt; protoplasm
coarsely granular, disorganized, also the walls.

Hyphae as wide as germinated conidia, few septa,
granular protoplasm.

Hyphae swollen septation reduced; protoplasm
coarse, disorganized.

Hyphae enlarged, protoplasm and walls disorganized.

Hyphae enlarged, protoplasm and walls disorganized.

Hyphae enlarged, protoplasm weakened, has clear
appearance.

Hyphae much weakened, protoplasm reduced.

Hj'phac slightly enlarged, protoplasm and walls
disorganized, septation reduced.

Some hyphae swollen, others shrunken, hazy appear-
ance; protoplasm badly disorganized.



26



Histological Features — Continued



Agents



Kind



Per

cent



Size of

germ.

conidia

in/x



Characteristics of development



Formic acid

Sodium formate. .
Saccharin

Salicylic acid

Sodium salicylate

Oidiiim
Alcohol

Benzoic acid

Sodium benzoate .

Boric acid

Borax

Creosote

Formaldehyde. . .

Formic acid

Sodium formate. .
Saccharin

Salicylic acid

Sodium salicylate.



0.2
0.5
0.3

0.05

0.05

5.0
0.02

0.03

0.2

0.5
0.05

0.1

0.2
0.5
0.3

0.05

0.05



Hyphae enlarged, protoplasm and walls disorganized.

Hyphae enlarged, protoplasm and walls disorganized.

Normal size, protoplasm non-hyaline, swollen ap-
pearance.

Normal size, protoplasm non-hyaline, swollen ap-
pearance.

Soine hyphae swollen, protoplasm disorganized.



Hyphae swollen, septation reduced; protoplasm
coarse, disorganized.

Hyphae slightly swollen, walls and protoplasm
weakened, break easily. vSome hyphae entirely
disorganized, protoplasm reduced, coarsely granular.

Hyphae slightly swollen, walls and protoplasm
weakened, break easily.

Hyphae enlarged, protoplasm weakened, has clear
appearance.

Hyphae much weakened, protoplasm reduced.

Size normal, but disorganized appearance, septation
reduced.

Some hyphae swollen, others shrunken, hazy appear-
ance; protoplasm badly disorganize(l.

Hyphae enlarged, protoplasm and walls disorganized.

Hyphae enlarged, protoplasm and walls disorganized.

Normal size, protoplasm non-hyaline, swollen ap-
pearance.

Normal size, protoplasm non-hyaline, swollen ap-
pearance.

Some hyphae swollen, protoplasm disorganized.



The results of the tests with these preservatives show the acid to be
stronger in all cases than the corresponding salt,^ and also that the benzoic
acid was the strongest antiseptic in its action on the three moulds. In
testing the preservatives in this group, the ordinary beef gelatin and
tomato gelatin were used as well as the tomato bouillon, but the gelatins
were found unsatisfactory on account of irregularity in development. The
gelatins permitted only a surface growth, which limited the absorbent
action of the hyphae to the part in contact only, whereas, in the soltition the
entire surface of the hyphae acts as an absorbent. The sodium benzoate,
as it was the most generally used preservative, was tested under a greater
variation of conditions than the others, and presented many peculiar
features. In some cases, the development might show a greater retarda-
tion, but less effect on structure, than in others where the development
seemed more nearly to approach the normal in extent, but on microscopic
examination a more marked effect on structure could be seen. In some
tests where the smaller amounts of ])reservative were used, the disorgan-
ization of both the protoplasm and tlie cell-walls in the early development
was as great as with the larger amounts, while the later development might

* E. L. Trouessart: Antiseptic Therapeutics, Vol. 1, 1893. Trans, by E. P. Hurd\



27



be almost normal, as if the entire effect were concentrated in the first
growth, or some colonies might show a marked effect and others little
or no effect, both developing simultaneously.

The Alternaria and Oidium were much more sensitive to the action of
both the benzoic acid and the sodium benzoate, the solutions in which
growth occurred being only about one-tenth the strength of those in which
the Penicillium was able to develop. Similar selective effect to that in the
Penicillium was also apparent in these organisms, some hyphae showing the
disorganization as in the Penicillium, while others appeared normal.

Disorganization of both protoplasm and cell-walls resulted in varying
degrees from the action of the preservatives in this group. The disor-
ganization was manifested in swelling and distortion, the protoplasm becom-
ing coarsely granular and without cohesion, there bemg no perceptible dis-
tinction between the limiting membrane and plasma, also in varying
degrees of yellowing or browning, and in ru])turc from the mere handling
in the transference to the slide, or the weight of the cover-glass, or the
change in osmotic action due to the mounting in a drop of water instead
of the medium in which grown. The protoplasm, when colored, had a
rusty, dirty appearance, not the clear color which is produced when stained.
When the wall ruptured, the protoplasm flowed out, the coarse granules
spreading in all directions, showing total lack of cohesive force. Buchanan^
states that salicylic acid has the peculiar property of macerating flesh, so
that the hands are attacked, when it is used as an antiseptic, and on
account of this property it constitutes one of the ingredients of "corn
salves." The other members of this group which behave practically alike
in their effect on the moulds, apparently do not possess this property,
though butyric acid which is also similar in its effect on the moulds has been
used by Loeb- to destroy the superficial cortical layer of the unfertilized
egg before development can take place.

The Alternaria and Oidium could not be tested in so close a series as the
Penicillium as their growth was inhibited more easily; in the one case a
fairly good development was obtained with a small amount of the preserva-
tive, but a slight increase above this amount completely inhibited develop-
ment. Very minute amounts of the preservatives were sufficient to pro-
duce marked effects, the effects being similar to those obtained on the
Penicillium. See Plates 20-33.

FIFTH GROUP

In this group are substances which have been classed as antiseptics, dis-
infectants, or as having an injurious eft'ect on living matter. Alum and cop-
per sulphate have been used in the preparation of certain foods, and the
other salts listed have sometimes entered foods from the vessels used in
preparation or from the containers, and are said by some writers to be
poisonous.^

For the tin compounds to be used in the tests the stannous series was
selected as these are the ones to which poisonous properties are attributed,
though in the government regulation'* cognizance is not taken of the form
in which the tin occurs ; the basis for the regulation appears to be the result
of chemical analyses, including both the stannic and stannous series.

^C. M. Buchanan: Antisepsis and Antiseptics, 1895.

2 J. Loeb: Popular Science Monthly, 80 (1), 1912.

3 y. W. Holland: Medical Chemistry and Toxicology, 1905; H. Leffmann and W.
Bearn. Select Methods in Food Analysis, 1905; K. P. McElroy and W. D. Bigelow,
Dept. of Agr., Bui. 13, part 8, 1893.

* Food Inspection Decision, No. 126.

28



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Many tiny sulnnerged

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Surface covered, dirty

appearance.




Many tiny sub-
merged.

Many surface and
submerged colonies.

Patches of film

Patches of film

Many submerged
colonics.

Patches of film. . . . . .

Hard white dots on
surface, many sub-
merged.




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30



Histological Fe.\tures



Agents
Kind


Per

cent


Size of
germi-
nated
conidia
inyu


Characteristics of development



Penicilliiim
Alum



Copper sulphate. . .

Ferrous sulphate . .

Lead acetate

Sodium sulphate. . .
Sodium sulphite. . .

Stannous oxide. . . .
Stannous p h o s -

phate
Stannous p r o t o -

chloride
Stannous sulphate.

Zinc chloride

AUernaria
Alum

Copper sulphate. . .

Ferrous sulphate . .

Lead acetate

Sodium sulphate. . .
Sodium sulphite. . .
Stannous oxide. . . .

Stannous p h o s-

phate.
Stannous p r o t o -

chloride.
Stannous sulphate .
Zinc chloride

Oidiurn
Alum

Copper sulphate. . .

Ferrous sulphate . .
Lead acetate



1.0


11.4
19.0


0.2


11.4


0.05


8.5


0.05


8.5


2.0
0.2


8.5
11.4


5.0
2.0


8.5
11.4


0.02


11.4


0.02


11.4


0.2


4.0
11.4


0.5




0.1




0.2




0.2
1.0
0.2
5.0




1.0




0.2




0.1
0.5




1.0




0.5




1.0
0.5





Large round cells are formed from gferminated

conidia from which are developed attenuated hyphae.

Fruiting heads developed close to germinated

spores, some of which are thin and small.
Hyphae enlarged, slightly distorted; protoplasm

yellow, finely granular, has dirty appearance.
Hyphae normal size, many small fruiting heads

formed.
Protoplasm coarse disorganized; some strongly

vacuolated.
Some hyphae enlarged, strongly vacuolated.
Hyphae enlarged, few septa; protoplasm coarsely

granular.
Hyphae attenuated, protoplasm coarsely granular.
Hyphae badly disorganized, coarse protoplasm.

Some hyphae enlarged, disorganized; some shrunken
with disorganized protoplasm.

Many hyphae swollen, ends break with weight of
cover glass; protoplasm coarse, some strongly
vacuolated.

Two forms of hyphae, one set large, regular, smooth
outline, coarsely granular protoplasm; other set
small, distorted, protoplasm homogeneous. Fruit-
ing heads small, branches in various directions.

Hyphae irregular, ragged appearance, septation in-
creased; protoplasm coarsely granular.

Hyphae slightly enlarged, walls and prdtoplasm
disorganized.

Some hyphae shrunken, septation increased; pro-
toplasm coarsely granular.

Hyphae empty, septation increased.

Hyphae disorganized, protoplasm coarsely granular.

Hyphae enlarged, protoplasm coarsely granular.

Hyphae normal size; protoplasm attenuated, coarsely
granular.

Hyphae enlarged, protoplasm coarsely granular.

Hyphae normal size; protoplasm attenuated, coarsely

granular.
Hyphae disorganized, enlarged.
Hyphae enlarged, distorted, break easily; protoplasm

coarsely granular escaping from hyphae.

H^-phae shrunken, septation reduced; protoplasm
stringy.

Some hyphae enlarged, irregular; protoplasm coarsely
granular ; others attenuated, disorganized prominent
septa.

Hyphae distorted, attenuated, protoplasm coarse.

Some hyphae shrunken, irregular; protoplasm at-
tenuated, many vacuoles.



31



Histological Features — Continued



Agents



Kind



Per
cent



Size of
germi-
nated
conidia
inM



Characteristics of development



Sodium sulphate.. .
Sodium sulphite. . .
Stannous oxide. . . .

Stannous phos-
phate.

Stannous p r o t o -
chloride.

Stannous sulphate.

Zinc chloride



2.0
3.0
5.0





3.0




0.05




0.05




0.5





Swollen, disorganized, homogeneous protoplasm.
Slightly enlarged.

Hyphae hav^e ragged appearance; protoplasm re-
duced, coarse.
Hyphae enlarged, protoplasm coarsely granular.

Hyphae attenuated, indistinct, septation reduced;

protoplasm coarse.
Hyphae normal size, ragged appearance; protoplasm

coarse.
Hyphae enlarged, irregular; protoplasm coarsely

granular



The Alternaria was the most susceptible of the organisms to the action
of the alum, both in checking development and histologic effect. Similar
results, though with different ainounts of the altim were produced in all,
evidenced in an impoverished-appearing protoplasm, the h\-phae appearing
somewhat clear and more attenuated as the amount of the alum was in-
creased to the inhibiting point, and the protoplasm more reduced.

The copper sulphate in the lesser amounts produced enlargement and
disorganization, the protoplasm being rather coarsely granular and lacking
cohesion. In the larger amotints, germination was retarded, the hj'phae
were stunted and, in some cases, misshapen. Clark, ^ using molar solutions,
found that N/128 (0.1 per cent) inhibited growth in P. glaucum, and that
N/1 was required to kill the mould. Pfeffer- states that P. glaucum is still
able to grow in 21 per cent. They undoubtedly worked with different
strains of what was known as P. glaucum. Alsberg and Black^ found that
species of Penicillium so closely related that until recently they were not
distinguished by morphologists, differ quite markedh- in their metabolism.
The writer found that in the development of the Penicillium used in the
tests the organism was more sensitive to slight changes in environment, as
shown mor])hologically, as well as being more resistive, as shown in develop-
ment when transferred to normal media, when the inoculations into the
test media were made from cultures which had been kept in vigorous condi-
tion through transfers made f airl v regularlv and at somewhat short intervals.



^ J. F. Clark: On the toxic effect of deleterious agents on the germination and de-
velopment of certain filamentous fungi. Bot. Gaz., 28: 28^, 375, 1899.

- W. Pfeffer: Physiology of Plants, Vol. 11: 60, 1906.

3 C. L. Alsberg and O. F. Black: Contributions to the study of maize deteriorations.
Bui. 270, B. P. I., Dept. of Agr.



32



The ferrous sulphate and zinc chloride in the largest amounts produced
enlargement and disorganization, the latter manifested by the rupture of the
walls and the free flowing out of the coarsely granular protoplasm. Pfeffer
states that P. glaucum is able to grow in solutions of ferric sulphate up to
8 per cent, which would indicate that the ferric salt was less poisonous.


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Online LibraryKatherine Golden BittingThe effect of certain agents on the development of some moulds → online text (page 3 of 7)