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1.
Me d i ap r e p a r a t i o n Mi c r o b i o l o g y N a me : H a b i b a j a mi l R o l l n o: 8 6 1 B s z o o l o g y 7 t h s e me s t e r P r e s e n t e dt o: P r o f . F a z a l E l a h i B u t t
2.
Me d i ap r e p a r a t i o no r C u l t u r e me d i ap r e p a r a t i o n: B a c t e r i a l c u l t u r e o r Mi c r o b i o l o g i c a l me d i a i s a g r o w t hme d i u m u s e dt og r o wb a c t e r i a . I n o t h e r w o r d s i t c o n t a i n s e v e r y t h i n gt h a t b a c t e r i a n e e d s t og r o wo u t s i d e t h e b o d y a n du n d e r l a b o r a t o r y c o n d i t i o n s . O R Mi c r o b i o l o g i c a l C u l t u r e me d i u mi s a s u b s t a n c e t h a t e n c o u r a g e s t h e g r o w t ha n ds u p p o r t s u r v i v a l o f mi c r o o r g a n i s ms . C u l t u r e me d i a c o n t a i n s n u t r i e n t s g r o w t hp r o mo t i n gf a c t o r s , e n e r g y s o u r c e s , b u f f e r s a l t s , mi n e r a l s a n dt h e g e l l i n ga g e n t s ( f o r s o l i d me d i a ) P u r p o s e s : T ob e c o me f a mi l i a r w i t h t h e n e c e s s a r y n u t r i t i o n a l a n d e n v i r o n me n t a l f a c t o r s f o r c u l t u r i n gmi c r o o r g a n i s ms i nl a b s . T ou n d e r s t a n dd e c o n t a mi n a t i o n o r s t e r i l i z a t i o np r o c e s s u s i n ga na u t o c l a v e . T ol e a r nt h ep r o c e d u r eu s e di np r e p a r i n gme d i an e e d e df o r
3.
c u l t u r i n gmi c r o o r g a n i s ms . T y p e s : B a s a l me d i a : A n u n s u p p l e me n t e dme d i u mw h i c hp r o mo t e s t h e g r o w t ho f ma n y t y p e s o f mi c r o o r g a n i s ms w h i c h d on o t r e q u i r e a n y s p e c i a l n u t r i e n t s u p p l e me n t s . O r B a s a l me d i a a r e t h o s e t h a t ma y b e u s e df o r g r o w t h( c u l t u r e ) o f b a c t e r i a t h a t d on o t n e e de n r i c h me n t o f t h e me d i a . E x a mp l e s : N u t r i e n t b r o t h , n u t r i e n t a g a r a n dp e p t o n e w a t e r . E n r i c h e dme d i a: E n r i c h me n t c u l t u r e i s t h e u s e o f c e r t a i ng r o w t hme d i a t of a v o r t h e g r o w t h o f a p a r t i c u l a r mi c r o o r g a n i s mo v e r o t h e r s , e n r i c h i n ga s a mp l e f o r t h e mi c r o o r g a n i s mo f i n t e r e s t . T h i s i s g e n e r a l l y d o n e b y i n t r o d u c i n gn u t r i e n t s o r e n v i r o n me n t a l c o n d i t i o n s t h a t o n l y a l l o wt h e g r o w t ho f a no r g a n i s mo f i n t e r e s t . Ame d i u mma y b e e n r i c h e d , b y t h e a d d i t i o no f b l o o do r s e r u m. E x a mp l e s o f e n r i c h e dme d i a i n c l u d e s h e e pb l o o da g a r a n d c h o c o l a t e ( h e a t e db l o o d ) a g a r . S e l e c t i v e me d i ac o n t a i ni n g r e d i e n t s t h a t i n h i b i t t h e g r o w t ho f s o me o r g a n i s ms b u t a l l o wo t h e r s t og r o w . Me d i a t h a t i n h i b i t t h e g r o w t h o f u n w a n t e dmi c r o o r g a n i s ms a n d s u p p o r t t h e g r o w t ho f t h e o r g a n i s mo f i n t e r e s t a r e c a l l e ds e l e c t i v e
4.
me d i a . S e l e c t i v e me d i u mc o n t a i np a r t i c u l a r i n g r e d i e n t s t h a t i n h i b i t t h e g r o w t ho f c e r t a i nmi c r o b e s . A ne x a mp l e o f a s e l e c t i v e me d i u mi s Ma c C o n k e y a g a r ( Ma c C o n k e y a g a r i s a s e l e c t i v e a n dd i f f e r e n t i a l c u l t u r e me d i u mf o r b a c t e r i a . I t i s d e s i g n e dt os e l e c t i v e l y i s o l a t e G r a m- n e g a t i v e a n de n t e r i c b a c i l l i a n dd i f f e r e n t i a t e t h e m b a s e do n l a c t o s e f e r me n t a t i o n . L a c t o s e f e r me n t e r s t u r n r e d o r p i n k o nMc C o n k e y a g a r , a n dn o n f e r me n t e r s d on o t c h a n g e c o l o r ) I n d i c a t o r Me d i a: A n i n d i c a t o r i s i n c l u d e di n t h e me d i u m. Ap a r t i c u l a r o r g a n i s m c a u s e s c h a n g e i nt h e i n d i c a t o r , e . g . b l o o d , n e u t r a l r e d , . E x a mp l e s : B l o o da g a r a n dMa c C o n k e y a g a r a r e i n d i c a t o r me d i a . T r a n s p o r t Me d i a : T r a n s p o r t me d i a a r e e s s e n t i a l l y s o l u t i o n s o f b u f f e r s w i t h c a r b o h y d r a t e s , p e p t o n e s a n do t h e r n u t r i e n t s ( e x c l u d i n gg r o w t h f a c t o r s ) d e s i g n e dt op r e s e r v e t h e v i a b i l i t y o f b a c t e r i a d u r i n g t r a n s p o r t w i t h o u t a l l o w i n gt h e i r mu l t i p l i c a t i o n . D e p e n d i n go nt h e t y p e o f o r g a n i s ms s u s p e c t e di nt h e s a mp l e , t r a n s p o r t me d i a ma y v a r y . S t o r a g e me d i a: Me d i a u s e df o r s t o r i n gt h e b a c t e r i a f o r a l o n gp e r i o do f t i me . E x a mp l e s : E g gs a l i n e . me d i u m, c h a l k c o o k e dme a t b r o t h . N u t r i e n t B r o t h: I t i s N u t r i e n t A g a r t h a t l a c k S o l i d i f y i n ga g e n t , a g a r
5.
p o w d e r . I t r e a ma i ni nl i q u i df o r ma t r o o mt e mp e r a t u r e a n da r e u s e dt oma i n t a i ns t o c k s o f Mi c r o o r g a n i s ms . C o mp o s i t i o n: B e e f e x t r a c t 3 g, P e p t o n e 5 ga n dWa t e r1 0 0 0 ml . N e u t r i n t A g a r : I t i s a g e n e r a l me d i u ms u p p o r t i n gg r o w t ho f a w i d e r a n g e o f N o nf a s t i d i o u s o r g a n i s ms . f o r e x a mp l e N e i s s e r i a G o n o r r h o e a w h i c hb l o o do r h a e mo g l o b i nw i t hs o me o t h e r r e q u i r e me n t s . C o mp o s i t i o n: B e e f e x t r a c t 3 g, p e p t o n e 5 g, A g a r 1 5 g, w a t e r 1 0 0 0 ml . P e p t o n e : A n y w a t e r s o l u b l e mi x t u r e o f p o l y p e p t i d e s a n da mi n o a c i d s f o r me db y t h e p a r t i a l h y d r o l y s i s o f p r o t e i n s . B e e f E x t r a c t : Mi x t u r e o f p e p t i d e s , A mi n oa c i d s , mi n e r a l , s o me v i t a mi n s , o r g a n i c a c i d s a n dn u c l e o t i d e f r a c t i o n s . B DB B L. B DB a c t o . A g a r : P o l y s a c c h a r i d e s i s o l a t e df r o ma p l a n t a n di s u s e df o r t h e Mi c r o b i a l c u l t u r e s o n s o l i dme d i a . s t e p s o f a me d i aP r e p a r a t i o n: Ma t e r i a l s : C u l t u r e me d i a ( E MBa g a r me d i a f o r e c o l i )
6.
C o n i c a l f l a s k w i t hc o t t o n p l u g N e u t r i n t A g a r f o r c o mmo nb a c t e r i a Me a s u r i n gC y l i n d e r D i s t i l l e dw a t e r P e t r i p l a t e s S t e p s : F i r s t t a k e t h e c u l t u r e me d i a a n dw e i g h t i t , a s w e t a k e 3 6 gi n 1 0 0 0 ml t h e nh e r e w e w i l l t a k e 3 . 6gI n1 0 0 ml . N o wp u t t h e c u l t u r e me d i a i n t oc o mi c a l f l a s k a n dt h e nme a s u r e 1 0 0 ml o f d i s t i l l e d w a t e r u s i n ga me a s u r i n gC y l i n d e r t h e np o u r t h e w a t e r i n t oc o n i c a l f l a s k n o ws h a k e a n dp l u gc o t t o n. I f w e n e e dt oa u t o c l a v e i t t h e nf i r s t l y b o i l i t o nh o t p l a t e . A u t o c l a v e : N o wc o v e r t h e mo u t ho f f l a s k a n dp e t r i p l a t e s w i t ha l u mi n i u mf o i l . H e a t a u t o c l a v e f o r 1 5t o2 0mi n u t e s a n dn o wp u t b o t ht h e p e t r i p l a t e s a n df l a s k i n t ot h e a u t o c l a v e f o r 1 5mi n u t e s . S o l i dme d i a: i n v o l v e s mi x t u r e o f a g a r a n dn e u t r i n t p o u r e di np e t r i d i s h e s B r o t hMe d i a : N u t r i e n t b r o t hme d i a i s a l i q u i dme d i u mw i t h o u t a g a r t h a t c o n t a i nt r y p t o n e , y e a s t p o w d e r a n do t h e r i n g r e d i e n t s . We u s e t e s t t u b e s f o r t h i s t y p e o f me d i a p r e p a r a t i o n.
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