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(1)

Bull.Fa c.B io r e s o u r c e s,M ie Univ.

No.3 3:1

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⁷

M a r ch 3 1, 2 0 0 6

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C hutim a K H O M VI LA I

*

, M a s a ak i K A SH IW A G I ^{a n}d M otoi Y o s HIO K A

"

Fa c ulty ôf B io r e s o u r c e s,M ie Univ e r sity,1 5 7 7, Ku rim am a ch i yâ^‑^cho,Ts u ,M ie5 1 4^‑8 5 0 7,Jâp^{a n}

A bstra ct

To e v alu ate the a nti fu ng^al a ctivitie s of ho r s e r ad ish Ar m o r a cia r u stic a n a e xtr a ct,the m y^{c e}lia a nd

z o o sp^{o r e s} ôf Sap^{r o}^lêgⁿia p^{a r a 5}ⁱ^tⁱ^{c a} ^NJ^M ^{8 6 0 4} ^w^{e r e} ^t^{r e a}^tê^d ^wⁱ^t^h ^{v a r}ⁱ^{o u s} ^{c o} ^m^{b i}ⁿâ^tⁱ^{o n s} ô^f âl l yl is oth io cy^{a n a}te (A I T) c o n c e ntr atio n a nd e xp^{o s u}r e tim e s^. A I T is a m ain p^{u n}g^{e n}^t ^{c o}^m p^{o n} ^{e n}^t ôf ho r s e r ad ish･ T he minim um inh i b ito ry c o n c e ntr atio n fo r m y^{c e}lial g^{r o w}th w a s6 8 mg / L ^with 6 0

1

^mⁱⁿ

e xp^{o s u r e} ^{a n}dthat f らrz o o sp^{o r e}g^{e r}^miI ‑ atio n w a s4 2.5 ^mg / L ^wi_th 5^‑^min e xp^{o s u r e}^･T he s e v alu e s w e r e s lgⁿi fic a ntl y l^{o w} ^{e r}^tha n tho s e r ep^{o r}ted in p^{r e v}io u s r e s e a r ch. T he ho r s e r ad ish e xtr a ctis a n effe ctiv e a nti fu ng^alag^{e n}^t ^ag^{a l n S}^t ^{S a}P^{r O}leg^{n l a S}is･

K ey W o rds :all y l i^{s o}^th io cy^{a n a}^tê^,^{a n}^ti fu ngâla ctivity^,ho r s e r ad ish e xtr a ct,Sap^{r o}^lê^gⁿⁱâ p^{a r a ∫}ⁱ^tⁱ^{c a}

Intr odu ctio n

A q^{u a}^tic fu ngi ôfthe o^rde r Sap^{r o}legⁿiale s ofte n c a u s e s e rio u sda m agê ^tô fr e shw ate r fishe sif no eff^Te ctiv e

p^{r o}p hyla ctic o r the r ap^{e u}^tic tr e atm e nts a r e ap pli^ed to the m^l)

･ M ala ch ite g^{r e e n} is q^{u l}^tê ^{a n} êffe ctiv e a ntifungâl ag^{e n}^t^,butitis teratogêⁿic^{2 )},re si du al^s⁾and c arcin ogêⁿic⁴)

. In

J

^ap^aⁿ ^,^the P ha r m a c e utic al A ffair s

La w p^{r o}h i b itedthe u s e of itin 2 0 0 3^･ T he c a r etake r e xte n sio n tim ef ♭r m ala ch ite g^{r e e n} had

j

^{u s}^t ^e^xp i^{r e}d iⁿ

J

û^ly 2 0 0 5^. A n alte r n ativ e a nti fu ngâl ag^{e n}^tis n e eded to be effe ctiv e a nd s af^Te fo r n o n^‑ta rgê^t ô^rg^{a n}is m s a s w ell a s the e n vir o n m e nt. C ur rent r e s e a r che s fo r alte rn ativ e ag^{e n}^t^s ha v e fo c u s ed m ainl_y on thre e c and idate s

,fo r m alin

‑

⁵⁸⁾

,h_ydr og^{e n} p^e^r^o^xi de

‑

⁶^‖),a nd s od iu m ch lo ri de o r s e a w ate r⁶

1

^{1 2}⁾I T he s e a r e effe ctiv e,but ha v es o m e n egâ^tiv ep^{o l n}^t^S^･ Fo r exa m p lê^,fo r malin is pô^t^{e n}^tiall_y ha r mful to the u s e r^'s he alth a nd r em ain s

in the en viron m e nt. R eg^{a r}d ing h_ydr og^{e n} p^{e r o}^xi de

, the u nd iluted s olution is str o ng ly c o r r o siv e and

c o mbu sti ble. A nd_,a s its effe ctiv e c o n c e ntr atio nis r eq^uir ed to be a s h i

g h

^{a s} ¹^,^{0 0 0} ^m g / L^,it ha s n otbe e n

p^{e r m}it ted in the U S A⁶⁾･ sod iu m ch lori de

,in sp^l^tê Ôf its s afet y, m ay be limited in its ap p li^{c a}b ili_{t y} du e to the h i g h ^{c o s}^t ôfa cq^{u l}^{r l}ⁿg êffe ctiv e c o n c e ntratio n sa s h i g h ^{a s} 3 0_,0 0 0 mg / L^･

U p ^tô ^{n o w} ^, ^w ê ha v ede m o n st^rated thats odiu m h_ァp^{o c}h lo rite ha s a pô^t^{e n}^tialto a cta s a n alte r n ativ e ag^{e n}^t thatis s upê^rio rin boths afety âⁿd affo rdâb ili_{t y t}o the for e m e ntio n ed che mic als^{1 3}

‑

^{2 0}⁾^･ Fu rthe r mo r e

, we ha v e

be e n try^{l n}g ^t^O ^{S e a r c}h fo r s afer ag^eⁿ^t^s^; ^{n o}^t ^syⁿthetic che ^mic als but n e a r^‑n atu r al s ubstanc e s･ U nde r the s e

p^{a r a m} ê^t^{e r s}^, ^w ê ôbtain ed the e xtr a ct of hors e r ad ish Ar m o r a cia r u stic a n a, wh ich is c o m m o nl y ^{u s e}d a s a spⁱ^{c e}

A c c ep^t^ed: M a r ch l O･2 0 0 6

* Pr e s e nt ad dr e s s; D^ep^{a r}tm e nt of F ishe rie s_, Ka s et

‑

^{K l}^{a n}^g^,^{C h}â^t^{v c}^hâ^k^,^B^{a n}^{g k}ôk 1 0 1 0 0,T hai la nd

* 辛 Fo r c o r r e sp^{o n}de n c e (ê^一^皿âîl^: ^m ô^tô^{i @ b i}ô ^皿ⁱê^‑û ^.^{a c}j^.p)

(2)

f

o r

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^ap^{a n e s e} ^{a n}d C h in e s e foods. Itis ^w ell kn o w n that _all_yl is othio cy^{a n} ^ate

(

^{A I T}; C H 2^‑C H ^‑C H 2^‑N ^‑C^‑S

)

,

being â ^m âin p^{u n}g^{e n}^t^{c o} ^m p^{o n e n}^t ôf ho r s e r ad ish, w a s ab i Wa 5 abia

j

^aP^{o n}ⁱ^{c a}^,^b^{r o w n} ^m ^{u s}^t^{a r}^{d B}^{r a s s}ⁱ^{c a}

j

^{u n c e a} ^{a n}^d

bla ck m u sta rd B･ nig^{r a} ^sho w s p^{o w}^{e r}ful ba cte rici de

2 1

1

^{2 3}⁾a nd fu ngi^ci de 2 4)

, a nd ha s als o be e n p^{r o v e n} ^t^o ha v e s o m e p^{o s}itiv e effe cts o n hu m a n he alth s u ch a s r e sp^{i r a}^t^{o r}y d is e a s e s

2 5)

･ He n c e, w e obs e r v ed the fu ngi^ci dal

a ctivitie s of A I T agâin stSap^{r o}^lêgⁿia p^{a r a s}ⁱ^tⁱ^{c a}^,â ^fi^s^h^‑pâ^thog^{e n} ^{o o m} y^{c e}te s, aⁿd det^{e r m}in ed both minim u m

inh i b itory ^{c o}ⁿ^{c e n}tr atio n s

(

^{M I C}^s

)

^f^{o r} ^m y^{c e}lial g^{r o} ^w^th a nd zo o sp^{o r e} g^{e r m}in atio n . In ci de ntall_y_, altho ugh M o riei al･

2 6)

alr e ad_yg^{a v e} both M I Csfo r the s a m e fu ng^{u s}^, ^w ^e ^cho s eto r ep^{o r}^t^{o u r} ^o ^{w n} ^{e x}p^{e r}im e ntalr e s ults wh ich we r e s lgⁿific a ntl_y d i ff^Te r e nt fr om their v alu e s,be c a u s e both c o n c e ntr atio n a nd exp^{o s u r e} ^tim e a r e

im p^{o r}^t^{a n}^tfa cto r sin s afet y ^{a n}d c o st a n al_ysisfo r p^{r a c}^tic al u s e of A I T a s a n a ntifu ng^al ag^{e n}^t^･

M ate rials a nd M ethods

H o r s e r ad ish e xtr a ct

A br a nd n a m e of ho r s e r ad ish extr a ct hot oil

(

^Yâ ^m ^{a c}^hû ^W ^{a s a}^{b i C}ô^.^L^t^d^.

)

^w ^{a s} ^{u s e}^{d i}ⁿ ^t^hi^s ^s^t^u^dy^. T he extr a ctin clude s 8 5

%

of A I T a sits m ain ing^{r e}d ie nt^. To p^{r e}p^{a r e}^the te st c o n c e ntr atio n sof A I T s olutio n_,the e xtr a ct oil w a s e m ulsi 丘ed to be a w ate r s olub le c o m p^{o u n}d,in clud iⁿg 8^.5% of A 士T_, a nd the n it w a s d iluted

with distilled w ater to e a ch ta rg^e^t c o n c e ntr atio n.

Fu ng^{u s}

Fu ng^{u s} ^{u s e}d in th is stud_y wa s a p^{u r e} ^s^t^{r a}in N

J

^M ^{8 6 0 4} ô^{f S}âp^{r o}^lêgⁿiaP^{a r a s}ⁱ^tⁱ^{c a} ^Cô^k^{e r} ^t^hâ^t ^{w a s}ⁱ^{s o}^lâ^tê^{d f}^{r o m}

c ultured c oho s alm o n On c o rki^y^{n C}hu s k is utch^. Pr ofe s s o r K ^.H atai, N i p p^{o n} V ete rin a ry ^aⁿd Anim al Scien c e

u niv e r si_{t y} k ind l_y s up p li^ed it. In o u rlabo r ato ry^, the fu ng^{u s} had be e n m aintain ed o n g l^{u c o s e}

‑

y^{e a s}t

(

^{G Y}

)

ag^{a r} ^{a n}d r efr e shed e v ery 3 o r 4 w e eks at5^oC ^{2 7}⁾^.

M y ^{c e}lt

'

al g^{r o} ^w^th inh i b ito 7y ^t^{e s}^t

Pr elimina^ril_y_,to de ci de the te stin c ubatio n te m p^{e r a}^t^{u r e}forthe fu ng^{u s},the ag^{a r} p lûg^s ^w ê^rê ^{c u}^t ^{o u}^t ^with

a c o rk bo^re^工5.5 ^m ^m in d ia m ete r 丘o m the ed gê ôf a ctiv eg^{r o} ^w^{l n}g ^{C O}lonァ,and _pla c ed o n the c e nter oftwo expê^rim ental G Y agâ^r p lâ^t^{e s}^,^the n in c ubated fo^r 2 days at 5,1 0,1 5,aⁿd 2 0ôC･ Co n s eq^{u e n}^tl_y_,the fu ngâl g^{r o} ^w^th w a s bet ter at the lat te r tw o te mp^{e r a}^t^{u r e s} ^tha n at the fo r m e r t^w o, a nd the r e w a s n o statistic al d i fferenc e betw e e n the lat te r tw o

(

^T^a^{b l}^e ¹

)

^. ^A^{c c o r}^{d i}ⁿgl_y_, the f わ^{r m} e r te mp^{e r a}^t^{u r e}, being ^{s u}itab le fo r s alm on _, w a s u s ed in th is study^. N e^xt the ag^{a r} pl^ug^s ^w ^{e r e} ^{c u}^t ^{o u}^tin the s a m e m a n n e r a s de s c ribed abo v e,

a nd _putinto l,0 0 0 m L of A I T s olutio n at c o n c e ntr atio n s of 6 8,8 5,a nd 1 0 2 mg / L ^with 3 0^‑ ^min a nd 6 0^‑min expo s u r e s. A fte r th is, e a ch plûg ^w ^{a s} ^rin s ed twic e with 5 0 m L ofste rile tap ^w â^tê^r ^{a n}d pla c ed o nthe c e nte r

Tab le 1 Colo ny d iâ ^m ê^t^{e r s} (^m ^m) ô^{f S}â_p^{r o}^lêgⁿiap^{a r a s}^{i li}^{c a} â^t ^v^{a r}ⁱ^{o u s}ⁱⁿ^{c u}^bâ^tⁱ^{o n} ^tê^m p^{e r a}^t^{u r e s}

5ôC 1 OôC 1 5ôC 2 0ôC

l l.5±0.2â 2 8.2±4.5^b 5 3.7±1.3^c 6 1.5±1.2^c T he d i ffe r e ntlet te r so ntheu p p^{e r} ^ri g h^t ^si de ofe ach c olo ny d iâ^m ê^t^{e r} ind ic ate a si gⁿi fic a nt d i ffe r e n c e betw e e n tr e atm e nts(^D^{u n}^{c a n}^'^s ⁿ^{e w}

multi p l^e ^{r a n}g^e^t^{e s}^t^,♪^<⁰^.^{0 5})