16

真 菌 と真菌 症

原

第4卷

第1号

著

Isolation

of Microsporum

cookei from Soil Attaching

Animal

to

Hoof

Shin OKOSHI, Mitsuo TAKASHIO and Atsuhiko HASEGAWA Department ofVeterinary InternalMedicine, Faculty of Agriculture, University of Tokyo The high incidenceof such keratinophilicfungi as Microsporum gypseumand Keratinomyces ajelloiin the soilof barns, barnyards,and otherareas,whichareinhabited or frequentedby domesticand wild animals,was demonstratedby Ajellol)and other investigators5,12). This fact was ascertainedby the present authorsin Japan6,7).These keratinophilic fungiare nowconsideredas soil saprophytes,the role of which is to decompose keratininto simpleelementsmicrobiologically1,11). These fungihave beenknownto invadehair and some other keratinaceoustissues of humanbeingand variousanimals4,8,9).The presentauthorsobservedthat they invaded the unguiarkeratinof variousanimalsin soilculture(unpublished data). Then, the authors conducteda survey to determinewhether these fungi were present in the soil attachingto animalhoofsand clawsor not. Duringthis survey, M. gypseumand K. ajelloiwere recovered. In addition,an isolateof M. cookei Ajello, 19592) was recoveredfroma soilspecimencollectedfrom the hoofsof a horse, which were free of lesions. According to Ajello2,3), this fungusspeciesis distributedwidelyin soil and is frequently isolatedfrom rodentsand otheranimals. Its presence,however,has never been reportedin Japan. This paperdealswith the morphological characteristics of the isolate of M. cookeirecoveredfor the first time in Japan. The resultsof the survey on such soilcollectedfrom the hoofsand clawsof animalswillbe describedin a separatepaper. Materialsand Methods Specimenswerecollectedfrom the soilattachingto the hoofsand clawsof animals by scrapingit offwith a sterilescraperinto a sterileglass bottle (Fig. 1). They were broughtto the laboratoryand examined for the presenceof keratinophilic fungiby means of a modifiedmethodof Vanbreuseghem's hair-baitingtechnique10). Baitingof a soil specimenby Vanbreuseghem's originaltechnique,usinghumanor animalhair as bait, oftenresultedin prevalentgrowingof contaminantmolds. Besides, the quantityof a soilspecimenwas, in somecases,smallerthan that requiredfor the test by this techniqueand could not fill half the capacityof a Petri dish9cm. in diameter. Accordingly, the following modification was madein the originaltechnique. A certainquantityof a soil specimenwas placed in a sterile Petri dish. To the

昭 和38年4月20日

dish

an

17

appropriate

kinds

of

soil

soil

of

with

tufts

placed

the

ated

was

well

as

strain in

was

the

This a

quantity

dish.

The

capacity

water.

of

Then,

of

horse

a dark

of

the

all

at

roam

cupboard

quantity

of

dish.

they

hoof,

both

Both

were of

baited

which

were

temperature

the

which

keratinaceous

was

chloramphenicol

morphological

baits,

suspected

as

a

agar.

A

characteristics

it

strain

on

was

examined

dermatophyte

was of

culture

M.

inocul-

cookei

media,

thus

grossly

as

microscopically.

This baited

for

the

the

shreds in

total

4weeks. on

fungus

of

kept

the

of

sterile

and

was

over

cycloheximide

examined

hair

dish

that

capacity

with

appeared

Any

so

one-sixth

horse

intervals

added,

the

about

and

growth

was

moistened

Petri

at

mycelial

Sabouraud

isolated

was and

The

microscope.

to

soil half

dish

human

examined

a

sterile

well

surface.

and

When

the

mixed

autoclaved

their

of

approximately

in

were

of

on

under

fill

placed

soil

(30-35•Ž.)

of

might

specimen

kinds

quantity

strain

horse of

of

this

were

examined

using

human

in

manner M.

which

premises

cultured

same

as

cookei

was

kept

farm,

soil

for

the

and

sterile

was

for

isolated

on

a

in

specimens

hair

as

at

soil

a

soil

Ibaragi

were

of

soil

natural

from

farm

presence

horse

moistened

described

room

temperature

specimen

collected

Prefecture.

gathered

At

from

keratinophilic

(30-35•Ž.),

specimens.

the

fungi

by

from

various surface

the

the

hoofs

sites of

in

soil.

hair-baiting

the They

technique

baits. Observations

By

the

tophytes

end

had

however,

on

the

growth,

molds

the

fungus and

and

macroconidia

had

thicker

mycelial

On

diameter

powdery

in

The

reverse

of

the

Under ores, were

downy the

were 35

to

a

Sabouraud

in

3 weeks.

zones

microscope,

in

and

and

of

10

were culture agar

surface

color, was

to

than

derma-

week

or

replaced

later,

the

continually

of

con-

overgrown

Its

hair

pyriform,

reve-

single-celled Its

macroconidia,

micro-

however,

to

Sabouraud

cycloheximide

it. temperature,

culture

was

aerial

growth in

(Fig.

4,

A

elliptical,

this pale

This and

had

attained

in

in some

color

zones.

pigmentation

showed

B). borne

multiseptate and

fungus

yellowish-tan

appeared

color.

macroconidia,

width

horse

macroconidia.

3).

room

the

margin

in

and or

inoculated

downy

were

human

clavate

reddish-purple

15ƒÊ

other third

gypseum.

at of

micro-and

to

the

(Fig.

M.

pure

peripheral

Macroconidia length

bait

echinulate

gypseum

fungus

dark

numerous

on

having

M.

woolly

wad

the

in

hoof

growth

dextrose

White

observed. 65ƒÊ

that

In

B).

those

The

culture

horse

multiseptate, of

molds

baits.

yellowish-tan

The

and

yielded

of

pale

mycelial

than

of

which

of

Microsporum,

those

texture. this

A

elliptical,

elements agar,

and

2,

contaminant

kinds

hair.

the

(2.5•`53ƒÊ)

Petri-plate in

through

was

resembled

chloramphenicol

8cm.

(Fig. of

experiment,

three and

horse

numerous

cell-walls

of

the

powdery and

examination

microconidia

The

week on

human

contaminant

that

second

prevalent

the

Microscopic aled

the

mycelial

taminants by

of

been

cell-walls

on and

simple

conidioph-

echinulate. 3

to

4ƒÊ

They thick.

Mi-

18

真 菌 と真 菌症

croconidia (Fig.

were

to

pyriform,

being

4

to

5ƒÊ in

length

and

1.5

to

2.5ƒÊ

第1号

in

width

5). The

than

culture

those

13

clavate

第4卷

to

19ƒÊ

in

These those

well

as

Survey

M.

the

was

isolated,

were

about of

grew

isolated

luxuriantly

which

were in

45

to

most

fungus

were

70ƒÊ

cases

were

of

in

(Fig.

quite

larger

length

and

6).

identical

with

isolated

from

none

of of

horse

horse

forms,

hoof

hair

such

No

fungi

sites

many

from

on

and

and

as

perfect

shreds, horse

those

stages,

as hoof

observed

in

however,

were

plate.

keratinophilic

various

hair

first

condial

reproduced. soil

from

horse

appeared

were

presence

on

Numerous

experimental

however,

macroconidia,

5ƒÊ thick

the

growth

hair.

specimen,

nor

collected

gypseum

was

soil

for

were

They

mycelial

human

natural

of

above.

fungus Its

on

hoof in

mostly

1959.

the hair.

natural

fromed

consisted mentioned

cell-walls

Ajello,

later

agar agar

characteristics

cookei

human

which

the

Their

culture,

and

the

on

width.

M. soil

shreds

oatmeal

morphological

of In

on

produced

in

them

in

the and

these

soil

a

large

number

premises K.

of

ajelloi

of

the

from

soil

specimens,

farm,

one

showed

that

M.

cookei

specimen.

specimens.

Discussion When horse of

the

hoof, horse

when

this

as

or

fungus The

hoof

was

was

M.

developed

cookei were

all

On

soil

specimen

whether

in the

the

with on

had

human

and

the

hoofs

collected, has

hairr

hair,

and

but

contaminant

baited

with

It

could

invade,

of

the

were

any

horse

horse by

baits.

hand, been

and and

soil

keratin

fungus

human

overgrown

autoclaved

three other

this

baited

continually

cultured on

shreds.

unknown

specimen

shreds

developed

positive

is

soil

fungus it

cookei It

the hoof.

keratins, well

hoof

to

not

on

molds. the

from have

no

effect

on

the

fungi

in

soil

of

shreds

However,

three in

horse,

found

pathogenic

shreds

kinds

of

vitro,

hair

as

which

this

M.

at

all.

lesions

hoofs

of

animals

not. Summary During

hoofs an

a

and isolate

This of

was this

claws of

specimen

survey of

been

the

first

the

animals

Microsporum

had

isolate

for

by

means

cookei

collected

of

of of

the

this

keratinophilic

a modified

Ajello,

from

recovery

were

presence

1959

was

hoofs

fungus

method

of

recovered a

in

of

Japan.

a

which The

to

Vanbreuseghem's

from

horse,

attaching

soil

were

technique, specimen.

free

morphological

the

This

from

lesions.

characteristics

described. References

1)

Ajello, 21,

L.:

The

157-171,

dermatophyte,

Ajello,

L.:

A new

3)

Ajello,

L.:

The

This

study

gypseum,

as

a saprophyte

and

parasite.

J. Invest.

Derm.,

1953.

2)

Ministry

Microsporum

Microsporum

ascigerous

was

of Education.

supported

and

state

of

its

occurrence

Microsporum

by a grant-in-aid

in cookei.

from

the

soil

and

animals.

Mycologia,

51,

173-177,

1961.

Sabouraudia,

1,

Scientific

Research

Fund

69-76,

of the

1959.

Japanese

昭 和38年4月20日

19

4) Dawson, C.O. and Gentles, J.C.: Perfect stage of Keratinomyces ajelloi. Nature, 183, 1345-1346. 1959. 5) Kaplan, W., Hopping, J.L. and Georg, L.K.: Ringworm in horses caused by the dermatophyte, Microsporum gypseum. Jour. Amer. Vet. Med. Ass., 131, 329-332, 1957. 6) Okoshi., S, and Takashio, M.: Isolation of Macrosporum gypseum and Keratinomyces ajelloi from soil in Japan and perfect stage, or cleistothecia, of M. gypseum. Jap. Jour. Med. Mycol., 3, 130-143, 1962. 7) Okoshi, S., Takashio, M. and Hasegawa, A.: Isolation of the keratinophilic fungi, Microsporum gypseum and Keratinomyces ajelloi, from soil in Japan. Proceedings of the 54th Meeting of the Japanese Society of Veterinary Science in Jap. Jour. Vet. Sci., in press. 8) Stockdale, P.M.: Nannizzla incurvata gen. nov., sp. nov., a perfect state of Microsporum gypseum (Bodin) Guiart et Grigorakis. Sabouraudia, 1, 41-48, 1961. 9) Vanbreuseghem, R.: La culture des dermatophytes in vitro sur des cheveux isoles. Ann. Parasitol., 24, 559-573, 1949. 10) Vanbreuseghem, R.: Technique biologique pour l'isolement des dermatophytes du sol. Ann. Soc. Belge Med. Trop., 32, 173-178, 1952. 11) Vanbreuseghem, R.: La vie saprophytique des dermatophytes. Ann. Derm. Syph., 87, 481-492, 1960. 12) Zeidberg, L.D. and Ajello, L.: Environmental factors influencing the occurrence of Histoplasma capsulatum and Microsporum gypseum in soil. Jour. Bacteriol., 68, 156-159, 1954.

抄

録

動 物 の 蹄 に附 著 した 土 壌 か ら分 離 したMicrosporum 大 越

伸 ・高 塩

cookeiに つ い て

満 男 ・長 谷 川 篤 彦

東京大学農学部家畜内科学教 室

著 者 らは畜 舎 な らび に畜 舎 の 近 傍 の 土 壌 内 に好 ケ ラチ ン性 真 菌-Microsporum gypseumお

よびKeratinomyces

ajelloi-が 高 率 に存 在 して い る こ と を既 に 報 告 した.更 にこ の両 種 の 好ケ ラチ ン性 真 菌 がsoil cultureに

お いて

useghemのhair-baiting

techniqueの

変 法 に よ り,好

ケ ラチ ン性 真 菌 の 調 査 を行 っ た. こ の 調 査 中,茨 城 県 下 某 牧 場 に飼 養 して い る1頭 の 馬 の蹄 か ら採 取 した 土壌 か らMicrosporum

cookei Ajello,

各種 動 物 の蹄 ・鈎 切 削 片 の ケ ラチ ンを侵 して発 育 す る こ

1959の1株

とを 認 め た の で,生 きた 動 物 の蹄 ・鈎 に附 着 して い る土

分 離 菌 株 の形 態学 的 特 徴 につ い て 記 載 した.な お我 国 に

壌 内 に も特 に好 ケ ラチ ン性 真 菌 が存 在 す る の で は な い か

お け る本 菌 の分 離 は こ れ まで 報 告 され た こ とが な い.

との疑 問 を抱 き,今 回 は こ れ らの 土壊 に つ い てVanbre-

を 分 離 し得 た.本 報 告 にお いて は主 と して 本

20

真 菌 と 真 菌症 第4巻

Plate

Fig. 1

Fig.

2A

I

Fig.

3

Fig.

2B

第1号

昭 和38年4月20日

21

Plate

II

Fig. 4A Fig.

Fig.

5

Fig.

4B

6

22

真菌 と真 菌 症

Explanation

of Plate

Fig1

1.

Collection

Fig.

2.

A.

horse

No

hoof B.

(left)

3.

a

specimen

growth

growth

molds

Profuse

soil

placed

Mycelial

contaminant Fig.

of mycelial

on

as

baits

of shreds

production

of

from is

seen on

M.

cookei

of

horse

one on

of

4.

A.

A

B.

Reverse

Fig.

5.

Sabouraud

Fig.

6.

Oatmeal

colony

on of

Sabouraud the

dextrose agar

slide

colony.

agar

slide

culture

hoofs hair soil

on

of

a

(top),

horse.

horse

hair

(right),

and

horse

hair,

and

shreds

around

culture

preparation. •~300.

agar.

a

human

filament

and

growth

of human

hair

by

M.

cookei, •~120.

II

After

preparation, •~300.

of

plate.

hoof.

dextrose

same

the

appeared

Plate

Fig.

I

autoclaved

macroconidia

第1号

Figures

human

an

第4卷

3

weeks

at

room

temperature

(30-35•Ž.).

of