1(Jift~ H~1R0' ~ 0 ~
.
.fiii
4 :
d1f~fF.
i7 • 1 :(l)m
~~m~1:~FEfl(8t!ij;tf'FFfl' tllilffi 1876 {f(nTyndall 1929${iJi*HJ.J (Flem
·Off
ing ) ~:fJ?:~.~
(
penicillin) (l), llli::fiJ 1940 tf::trffH~ffl!fH.tE~tltJ:' .tEm-=.*IT(8~:::':z
tit
w
*
!ilt
If~ ~~~t;5 A. 1m ~ , t:fJlt
titW
J:(8 f4 {~~1VtJE
t;t1: (81fiil ~{I£ ~ tfJ,~00
1940 iflf!f Waksman .tE± ~r:p %I~!hl:\g'E/.ttt1:
ttJ1i\[fv~ (Ac t inomyce te s )(2).Jlt 1&t1i
!.t~f¥J
'fEiE
A f&
urlDE :tJvt
(
Ant i b i 0 tic s ) - M: r±l ~!J=. ~tt1'J m
-=. f~ ~~1tW ~(Secondary metabolites ·::,lF~~t~1::f?:J'JT jj{Jt\h'lJi) (3), ttiW~!hT
rf)] flj( m~ ~gm ~H~~Q~*r±l~fi~~ ~·A~e ~~ .~r:p~. .r:pft~~tt1:*' ~m. M~~~~ ,:;%NJt .~ ~,~~~& & E(:J ~Jt
lEIJlt-
~ '~lt1M
~'~ m~,f1
j(oifij% 1i\[~H~j.[ ~i:!.l'::--mft ~ XV) ( S tr e pt omyce s) fr5J&~ 0 1~~ ~
1Vt
1'Gfl
0 JJII{Ill
'# lfx?J·~ ~.ftH ~~~~ r:p ~ ~ AjP~~AI!tt~1i\[~,,~~-~**I~j(r;J2f~*ffjJ 8 ' .r:tJJI: 1961 tflf!fWaksman (5). fi~
DoB
~1:%.' ;Jti,%i~tnj::lftfU1ft~m1:* ' (t,m: ~{~~~Streptomycin ~M# :
Actinomycin~.~ : Neomycin) ,~JlttJ Waksman ~ "l&ffA~Zx'~ ~j®UE !
~Nt M ~qJ tti~ ttl
*~~:!.l'::¥~mJE.AAmt1'J ~.~T,t:f
-~~0
tt1:
m~.~1VtJE ~'~7 ii-UJ1ft 51 • lk
ltff
JE
mit
1:
@ ~t1'J
if:;@ &11:
1t
-'f
~
it
~t
bl
7,;
*
1..**~!Y:J~
(l),*:f?:fflJ;~~1jl5 (
till
ilU\:Vi'H~:£· Glucose- nutrient agar medium) B~ ( Peptone )101;; •1ru1m
(Glucose) 10'l:: • TA\ft;ilrlj ( NaCI ) 5 'l::fl ' ~ ( Mea t extrac t ) 5'l:: • ~ (Agar) 20'l:: • *~*(
Distilled water) 1000 'iWGJ3l!:(pH ) 7 . 2 7 . 4 0(2)tf{ i£'JHj\; (~'HJIZ mig • Glucose-nutrient broth medium) R~l\[ ( Peptone) 10:L'::fl' 1ili.~ (Glucose) 10 {t~~ (NaCI )5 'l::
,l:fr;I~ (Meat extract) 5'l:: ,$;~*
(
Distilled water )l000 'M'~ 7.2 7 . 4 09Jfl1J
D±
715J1I¥
1lh
63 ~j;t~t t'tj:~ tr~ fttt iH 'fit'ffl
iIW:ii
• :j1'lfW3
&ffil 5(R fl':J. %~ • E~,fj!( ·f!f\ tr:J~ fE (5J f;j: '~~Are
.r:p%.W
(3) rnfli.' iJWl~ ~n~#4ffiJ ( Glucose-nutrient agar slant )~, R
7( , gl
f~ • f£'ftJi:
tr;-a-
100 ~ 1+~ rm ¥If:J 5001+ -
r~~lti $ ,tifti
27 T,5. ,fIJ J'l
J:£Ht
5 f~'!IX 15 lIH)Jlj-a-
2 50 ~1+ ~ttf 2000 ~Jt1i$ , ~fIDu~ Em ~ ~, 5~~~!IX' .~~~n,~~~~.~ 0 tf!fiB (4
)m
1:
t~'It
81trHt :
.~lf:J lei.l000~J+'~~~!t~3 4, jf!1J frlit1f:.~(Charcoal)t:p ~t*
, 0 ~~~(11+/
0~/~.),~ ~~~ ~ ~~'fflMlei. l.m~ ~1J~f~' J/1500 1!lW71<t5t il'ilfi:$ttJ¥ftrt ,IITd~0~'rt'/:80 %Efl (-2:2 %~
ttf~lei.
7/<) ~I±\ ' lita{f.pl"it~!&:!t (Fraction collector) *~
,
~ff
i~ .tg
vCfli1tffl(f>][~tfi:!&'*'
ilUfi'i:!:
M{~B..t6~~$ ( Lyo~hilizatin) ~rrXm~ftf~~~
o
DO
2 . .tt~~
(llffi~5::1~~H'tM : Staphylococcus aureuS ATCC 6638 Bacillus subtilis P.C.I. 219
££;~t1f:.M: Escherichia col i NIH] ,Pseudomonas aeruginosa
J.
ttl±:
~f.Jl
~ : Candida albicans YU 12000
(1)*m~ Nt 01:li,{!!!; , }iBt~Jfii~ m'±~ ~f1:
'
~J1l.ff
7f1:
(~t6* - ) , lit :1:1:12NH.,C ,{!!!; ,
JtBFi
m~IE
m
'±~If:J
m
0(2) E8 ~ -=.i
(2)mM iitU : :lifdl£iW)jjIJi! (Minimal inhibi tory concentration
,t1f:.§ M.LC.)0 tW~ ~ij~j:tf~£, Sabouraud's~,~~ £ , J:~H~ftfJ:
(3)m~
]:
£, #itillHl'fm~?l~
(
serial dilution) o.tt~Mlj-§5t4*'{!!!;W
~7/< (( ChI ( Normal saline )$ , fSJ~ ,{!!!;1J~ ~m~
IE
81~·ftl!ftf £1: 4.M~ttJ~n ~~ Mftf~, ~~~M1:~M~ttJ~. 0 E8=,
n
J.**
Mi1E 1:Z. tJL!£
~11'((J 't4= ft : ~lJl..tH
(l).i\lff;-e "ittFf ( Paper chromatography)m'±
'pj.:Ektrl,§,ir.ili~.~ (WhatSpiral
man 3MM)1:' "it7jIJ.Pj20%NH.,CI ,10%NH.,CI, 5%NH.,CI, 2.5%
5.
£ii**
Ma~N H ., C I ' 1. 3 % NH., C I ' O. 63 % NH ., C I ' 0 . 3 2 % NH., C I H20 f#I,
*
it~I±\~~S;(~'~ ~~ ,JfxA~ ]j'l:,f!j! (Bacillus subtilis ) 811m
tW
~ ),f!! ~ IiU~tr:J
nn
$ , I±\ ~iW ~ ~Rf fu!i 0 E8 Rf jEff it ifli~ , 1'imUMrr
,fiJ fi~)fr {i1J~o
fflbR~.~ (2)]fX.tt 9 • ~l.lUllDA3~ft·tJL~~~'.Pl~M:( HCI) :¥l:ft~( NaOH HJ.lJM:
Mlit~~J~
~
if
(:t(::9z
~2. J. 4. 5. 6. 7. 8. 9. 10.~ 10 0 0C
m
71< $ JJ[] 5 ~. • .lL ~tl i'l11;jl ~ ,J/1 NaOH, HC I i1f;lJj~ @[f 7.2· JJI]~~7l<~ 6
fH
lit ~ 2flim
)
o.Pl* ~-
'\ i;
1JD~~z.
lei.lJl.. 4fg:m~lei.~ trZ.P}liru (~) mJl.p.~(iCYlinder-platemethod ~JEm1:~t-.t ~~ 81f.!
'fifO
4. M
f1:
ttJt1iI* :
(1) ~M!ftf1t¥(~fH~$ftf~~F'G'H'I·]
KlJl..L)
f.tt~~ (Slide) 1t( StaphyhCuI ture ) , ~~ i~$ 14 • lIE.
m
~JJt 1:' .tlXT~~f:T~· 0 Staphyl(EflMtr!jj 15 30 '.P1Giemsa~-e$~-E· 1!liI.~~Y1:f~~Jm ( Balsam) Sacinia
11
• #it.fIX
*=.
~ Lt:~ ~Jj~T(
Jfx:k
125fli )
11~11B ~lJl..11B TH'J~ '*/J\ Bad 11 us, ill'.
-'f
J~ij 1* 0 ProteuslJl..11
14. Escherici I o )1:' ft ,-!iN P , ~*ff ) i:fJ f1jm~ 2%~ ~~:fi !FJXm~ )sa :fl :tt:12 ion lm'f~.
(7Jd
~~ 1: What !.5% JIUf:l ' J:00 Ii
~ , l'ffii1l!J ti (l'
Qt ,am)x'J\
!~14. 5. ~~;fIJm :
~~ ~~~~~ ,~~ ~m, ~T7
M~' ~mM%'~~.IIUR~ ~~~~~.~D~o~
~~*-:
1. ml£.11
{! f,liHiU~ ~t(:J ~ft : t:g*~L8~~~mO~~7.4'~- m7.2' .:::. ~5.8,m=-;R 6.8,lIt f& ~.II'!f Fs~ ttlt ftiTIi
~ ~ 1: 3t ' ~f\ ;RH:tf ~ 8. 8 0~ - tB
*
ml£11
;R ,m
tJ1J11
11'HI~J+,
1i ' " ,lIt~ II'!fM~ttlt~iTIiT~'~ ~._o2.ttl£~L8ttll~ :
E!3 ~
,PI W
:$:m
1£
~tI
Iii
.f£: ~fi
~!If Et:J jW iffIJ11 '
tI
.f£:~ tlJ: II ( (m.~03
) fllJ7'C ~ri
fi
jW iffIJtll§11 '
tI
m (191J :
Cand ida a I b i ca ns )tE.
& )#IJ11
0J·ttl£ ~ tI~'~~ ~L8 ttfEtlJ: :
(I)~~ NH 4C I f:!it~i:ffl~'
*
~L8 R f1W:~ Ch I0 rampheni ca I 1[lfJ?{ ( J2L Rf 1W:WNH 4C I
tI
~ftlfip} ~ili:*~~~:J~~ ) ~~_ 0(2)E!3 =-i1J~w' :$:ml£~:tE~rPj~~,~T~)J\ fi{*R':Jtt
3'&*,
~7f;~tffiH:tErp'~f, ~ ~~jrlJ:T~H~t! 0
(3)ttl£ PI~~Methanol (q3~) ,Ethyl acetate ( ~z:..H~),7.I<RCHCI3
(Chloroform' {jj) ,~m~ether(z:..~)rp;$: m~m~ 0
4.mnL8%~ :
E!3 lfi - , PI
w
:$:gMt:) ~ II - £;!I!*'*
Jfil Co n i d i a ph0 r e (~ff!H{i; ) ,fi
g~ %R~tt%.:::..f!f! '~*L8m
1. 5umx O. 5um , Ij\L8 ~O. 5umxO. 5um 0fi
Spira I 0 ~.~m:tE .f!f!~ :tE**tr~.~, .~) ,W~. ~ffl~.
•
ffl*
,~~~. ~ ,~ ~~. o~1ltPJ~:$:M m~illPJ~ilBfFm.i1rji, ~~filfU¥~fJl
(
Urease) 7tWf*,*f#::fiJm 0~ ~L8I£~~~RtI.~~ffiL8M
#
~tJ1J'~1ic ,£;!I!UmL8I£~'~~ ~EI3~H~(m~m:~H'~ (~~tl) ,~.~*H~o .@D~L8~~~W*,.U
¢~~~,~• •
,~~~,~.~~ ¥~. ~•
Staphylococcus aureus Staphylococcus epidermidis Sac i ni a 1 u t ea Bacillus subtilis Proteus mirabilis Escherichia coli ..-..--~.... ~--64>
1000 8 16>
1000>
1000 --13 -.~
'ps eudomonas aerugi nosa > 1000 ft~
K ebsiella pneumoniae > 1000 W~iU
Enterobacter::;2-' > 1000
Enterob~<:ter cloacae >1000
obac te r s p. > 1000
Candida albi ns >1000
l£10~ Ii ~0Sabouraud's
HlH\'ttJi:
((~ift.1j 0tfr/tIU2J.Ii ~~ tt(2*~ ).A~~OO~'~~AB (r~ ~N! 0 if,fH~ 37°Cg~$g~mgJ.tfr1t-~ ;Wj* 0 40 30 20 9 8
-
.
7 \ \ \ \ \ \ \ \ 6 \ / ' \.'
/ ' 5 4 ' - - - - + - - - I - - - - · - + I - - - - I - - -..., · · · - -_ _- - - + - - - + - o
1 2 3 4 5 6 7 8 ftffllJ ~~;YJ
0,+ ~t:j:Jft:lli[H!¥( Peptone) 1003[, .1@ 100·jHtfWl5
,rfr;:j
50R>
~r£7.2¥7.4o:(f T 11iR ±g (11iR ~ 3 0 71 ' ~Ji:
¥
1t
iii
60),':E
~JJ
f~ YJ~ ~zpt&)t t>J;WOOlfiiJ~
,
~ ~ YJ~ ~ ~~ 0 ...~~ fl(J1~ it--111
Rf 20 0.80 10 0.82 5 0.79 2.5 0.80 1.3 0.80 0.63 0.78 0.32 0.79o
0.80~ *,!!:l~ :(fiJ!l*ILt ( Wha tman 3 MM ), :lttif! :;:KmJ
iI
Bt (J1 NH 4 C 1 n;.lfX~ fJl3f~ , ffl ~ (Baci 11 us subt i lis) ~~Jl.
tem
:lXU~ , MIJ Rf 1tH: 0~~ ( pH1lft) 2 23.5 3 23.5 4 24.0 5 23.5 6 24.5 7 25.0 8 24.0 7 8 9 25.5 10 24.5 3 ~~ 1JL~~'.ef' D NaOH. HC 1 pH 11< 2. 3.4. 5. 6.7.8, 9.10.
am*
r:j:JJJn~ 5 1t~, W' pH 111'BI;SJ 7.2 JJom;j&*~ 61t,
D5TUJO ~~]!IUJ11~~lfX&4ffiW lfX~~~~D~~ ~~~ JJ~o
J A~'
• •
~A~.*4~±~~~'~••
'~~~M.~~ii!
*'%
,t!
@.. t~~*(!)~~ ¥; ~:£jH~% 1f.l. M~t~OO** ~t:fH~-B .fIi: 0 ~•
**
.. ElJttClllH
~%w
is) B-B-+@<~tiL
m
11
f'x:
Iff ,~~~Jjl (e) fS( ( t t )B-B
±
1m
tJLl£
a}j~ (D) fS(~B
B-B
tf--B
0 .t1tJ~. t tj::~ IE) fS(~1tJ*1t
~(
~< :tE~@'mi§U ,
~fl.
~llHiJ
(F)B
-+t3:t~1m
~'5ttJ1AA:ili**
:ettll
;$: ~~~P~~gJtiotm~ (G)f:E
B-B
~1:':.8-B
'fflt (H)1:£
B-B
~ ~ijttirJ:!*"* itHiJ
iJ~~m:Jl't,;~ n~ (I)
B1:':.
1m
;$:Ji~m~~ 8fl~
-itHiJ
(J)1:£
B
±~e ~tf- FL~ffrfi ' ~jl&
tJ1.ti: HiJ
(K) fS(~B
-iJ},u-B
@<~1:':.1m
TItJ~:tzt(;
B!l'- -
iJirJ:!
fJ: E~ (L) 1;:tB-B
m1:':.
tf--B
~-fit
~B.. C.. D-E 7}~AilA>g{&i~ :~. F" G.. H.. I ..
J ..
K" L11€ITll
' iiUl,:tE~ Til~ 147(0 Medium
ii!
*''r-Cal
eNf
KH
flP,ltilllt( ~~¥A) ~-t
·fB
*tili
**
t% KzMg rJ1I ~ ~ f;:t (
B
-~~1:':. ~~13 Ctiii
~ f;:tB 13
±
FE
!¥t ~ fS( ~B
8 -B
Ml ~ :J;fB-B
-±
±~13(¢*~~1:':.)z·
!ft ~1:£
~~1:':.B 1:':.
A III ~ 1;:t @<~-BB
D ~ ~@<
8
it*1t
~ Carbo!rtB
~ fi (1* )
i3:t~13 (l)Na f-Hi~Uft ( :;;r~ 15;-wx
illlt ) fS( ~13 (2)~~it;
(3)it~ --16 -~h.. • ifIIIl •
•
~• •
0.~' ~moo~~~% ~ ~.~~.®~ g~• •
®~. ,t1f~j*@J£cjJ
~(9)~~ 0Fi]TP1*3t]m
0fn~[r';Jj1f*4, ~~1f~[r';J.**.~1f[r';J;go
El3VtrfJ.
J:~*:$: rfJmi¥lt((JVt~~, &1HIH-}((J VtrfJ
,'8f.t
11ti¥rIl£f:;~£f:;.~ Vt.h~~,f.t~M£f:; .&~.[r';JVtrfJh~ ,m0~~~ffi~((J1f.f.t~oVt~~((J. &~re:$:*~~gJ£'~J:~~~Vt.hffigJ£.~~,ti~ ~~
L((J IJHiff: 0 tJSJ
tlt :$: 1C
lJlit?~&1'1
{Itu:e;rn
~*6 ,{g:fHiJi.
it ~ , t!@ ~+
7tff
~It
((J*l
(!l?;,~JItt 40°Cprf) , 0!J!BHiL
.hB':J~{~ 0 tt~@• •
~WcjJ(~fi.~~fi~~) ,~0ttWm• •
~=@~m.(~. ~~,~. ~) ,~~~= ft~,g~~.~m.'~~~K• •
(~Wm)El3
~~Vt~~~ ~Vt
~ ~~ ~* ~~ft=~7t~.~~[r';J~7t ~m*-
~~~Vt~ 0 .'8~~~~~'R1'1 M~-~[r';J~~oIt
lM
*
~~0%~'~M~. ~((J Bfi.,atlta0.*.tto~~.$~ ~~~, ~~~M~~M*4((J M,.~~~~m~[r';Jm.'&EmMM~M .~ f~~~ ~Mo~~~~~~m. ~[r';J • •
~~~~o~~.ft[r';J~mw ~(t:J-4l'JW
&
IfJ:[r';J
~WJ 0Medium A: Carbone nutrient medium
C~rbone source 1.0 gm (NH 4 ) 2S0 4 2.64 gm KH 2P0 4 2.38 gm K2 HP0 4 5.65 gm MgS0 4 • 7H2 O 1.00 gm CuS0 4 -5H 2 O 0.0064 gm Fe SO 4 - 7H2O 0.001 gm MnC 1 2 - 4H2O 0.0079 gm ZnS04 - 7H2O 0.0015 gm Agar 15_ 0 gm Dis till ed wa t e r 1000 _0 ml
Carbone source
ffi
(l)NaoAc.glucose ' IiJw ~
Jt'87tft]
*~~ihlilf2iFiHii
0(2):ft~'~3f=~' !}t~·N1t ~'*1~ millipore ~ f&~}J[]AB*.lli~.[r';J
£cjJ ,
ifiH~ ~i1! ~ 0(3HfMfl,
*
tmilI*E:ft
~ftilRIUt:J£cjJ
121°C, ~~m 15.~;M! 0
Medium B : Melanin Yeast ext ract
L- tyros i ne NaCI
Agar Tap wa ter
Medi forma t ion medi urn
( 1.0 gm 1.0 gm 8.5 gm 16.0 gm 1000.0 ml
Medium C: Potato plug
,~~
JlQi.
7J ~1JnXMIt
~§l ~iID81%il* ' 0. 10 %tt
1EE
~il 2 4 ~ 48 'N'if 1~, lli
ir?-tbt'§' cp Medi *~mal
21 &t ' r§] I!! ~ ~ 15 -5t~ 0G
K Medium D: Gelatin medium
M
Gel at in 160 - 200 gm
Tap water
Med i urn E : Mil k Skim mi lk Di s till ed wa t e r Me d i urn F : S u c r 0 sen i t r i t e Sucrose NaNOs K 2 HP0 4 MgS04 • 7H20 KCI FeS0 4 Agar Distilled water pH ; 7.0 -7 .3 1000 ml N F A D 150 gm 1000 ml Medi C T
c:
agar ( Capek's agar)
K 30.0 gm 2.0 gm A~ Di 1.0 gm pf 0.5 gm 0.5gm 0.01 gm 15.0 gm Medi 1000.0 ml Gl P€ Ag Sucrose 20 05U~ir?-30 ~:tt-?kcp~J1H@millipore fflf~j@i1I' JJIJA±gJt:£CPlt't:~,\)l
Di r§]~~~81nX-5tcpo
pH
\
..
Medium G : Glucose-asparagine agar
G I uc 0 se 10. 0 gm
Urea 2.0 gm
Aspar agine 0.5 gm
KzHP0 4 0.5 gm
Mar 15.0 gm
Dis tilled water 1000.0mI
PH: 6.8
1210
C •
~
I Medium H Glycerol-urea agar
Glycerol 15.0 gm KzHP0 4 2.0 gm MgS0 4 0.5 gm NaCI 0.5 gm FeSO 4 0.1 gm Agar 15.0 gm
Dis till ed water 1000.0 ml
Medium I : Glucose- tyrosine agar
CI u cose 1 O. 0 gm
Tyrosine 1.0 gm
{NH4 }z S04 0.5 gm
K zHP0 4 0.5 gm
Agar 15.0 gm
Dis ti lIe d wa ter 1000.0 ml
pH: 7 .0
121°C, &U~~
Medium J : Glucose-peptone agar
Glucose 40.0gm Peptone 10.0 gm Agar 20.0 gm Distilled water 1000.0gm pH: 5.6 -19
Medium K: Starch agar
So 1ubI e s t arch 10.0 gm 7.Waksman.
NaNOs 1.0 gm I dent i
K z HP0 4 0.3 gm Wi 11 ial
NaCI 0.5 gm 8.Waksman
MgCO s 1.0 gm ldenti
Agar 15.0 gm Wi 11 i a
Dis till e d wa t e r 1000.0 ml 9. Arai • A.
Toppa
Medium L : Potato-peptone-glYcerol agar
Peptone 2.0 gm Potato 100.0 gm Glycerol 5.0 gm MgS04 0.5 gm Kz HPO 4 0.5 gm NaCI 0.5 gm FeS0 4 0.01 gm Agar 15.
a
gm Tap water 1000.0ml B nH 50a
~*m;m7J<~~,
W
o 45n~'ill,
}JD/\ft~int~liz.m;m7J<'
121°Cl.Fleming .A.1929 A. Br. J. Exp. Pathol 10:226 2.Waksman.S.A. 1962 The Actinomycetes, Vol. 3
Antibiotics of Actinomycetes. Williams and Wilkins. BaItirrure.
3. Demain. A. L. 1972 J. Appl . Chern. Bi otechnol
22 : 345
4.Arai .A. 1976 The Actinomycetes; The Boundary Microorganisms
Toppan Company, Tokye. Japan.
5.Waksman • S. A. 1961 The Act i nomyce tes. Vo I. 2. Classif icaticn.
Identification and Description of Genera and Species. Williams and wi lkins • Bal timore.
6.Waksman.S. A. 1961 The Actinomycetes. Vol. 2 Classification. Identification and description of Genera and Species.
Williams and Wilkins.Baltimore . ... 20 ...
7.Waksman. S.A. 1961 The Actinomycetes. Vol. 2 Classification. Identification and Description of Genera and Species. Wi 11 iams and Wil k ins • Bal t imo re.
8.Waksman. S. A. 1961 The Actinomycetes. Vo1.2 Classification. Identification and Description of Genera and Species.
Wi 11 i am sand Wi 1kin s • B a 1timere.
9.Arai • A. 1976. The Actinomycetes; The Boundary Microorganisms. Toppan Company. o al tirmre. anisms atioo. ltion. -., 21 -.,