الأحياء مجهرية العملي

] [
] [
8241
1
2
3
4
5
6
4002
7 .........................................................
8 .................................
11 .......................................................
12 .................................................
. 12 ............................ Hanging drop Slide
. 13 ......................... Normal Skin Flora

15 ...........................
16 .........................................................
. 16 .....................................................
. 17 .................................... Simple Stain
. ( ) 21 .....................................
-1 21 ........................................................
-2 22 .....................................................
-3 24 .........................................
7
-4 ( ) 25 ...................................
28 .......................................................
22 ....................................................
. 22 ...............................
. 22 ............................ U-Shaped tube
. 22 ........................................................
( ) 31 ..................
: 31 ..................................................
] [
: 36 .................................................
: 41 ................................................
41 .......................................................
11
44 ........................................
12
47 .........................................
11
13
14
45 ...................................................
42 ............................................
52 ...........................................
- 52 .....................................................
- 55 ..............................................
58 ..............................................................
15
16
52 ....................................................
61 ...................................
63 .......................................................
- 65 ................................................... IMViC
- 62 ............................................
- 71 ....................................................... TSI
- 72 ............ Gelatinase
- 73 ....................................... Oxidase
-
74 ........................................ Phenylalanin-Deaminase
- 75 ..............................................
- 77 ...................................... Urease
- 78 ...................................... Catalase
- 72 ...................................... Coagulase
- 81 .................................................
- ( 82 ...... )Kirby Bauer
- 84 ..................................... Reductase Test

- 85 .............................................
17
18
] [
- 87 ...........................
21 ............................................... References
21 ........................................................
] [
21
22
2
4
22
23
25
26
11
28
12
111
5
6
7
8
2
11
13
14
15
16
17
18
12
21
21
22
23
24
25
26
24
25
27
22
111
111
112
113
Bacillus cereus
112
114
Clostridium tetani
115
117
116
117
118
Klebsiella pneumoniae
112
118
112
] [
27
111
31
112
32
112
113
22-28
31
33
34
35
36
37
38
32
41
41
42
43
44
45
46
47
48
42
51
51
52
53
54
55
111
112
113
114
()lowenstein - jensen media
114
115
115
Streaking.
116
117
118
112
()
121
121
122

Brewer
Micrometer
116
117
118
121
121
122
123
124
123
125
] [
56
58
57
52
61
61
62
63
64
65
66
67
68
62
71
71
72
73
74
75
76
77
78
125
.MR , VP
126
127
126
127
IMViC
128
128
122
TSI
122
TSI
122
131
131
131
133
135
) Bacillus subtilis )
137
138
API 20E Pseudomonas aeruginosa
132
134
136
137
138
132
] [
-1 .
-2 .
-3 .
-4
.
-5
.
-6 .
-7 .
.
-8 .
-2 .
:
-.
- .
- ( ).
- .
-.
-.
- .
] [
Microbiology
Microorganisms
: bacteria
fungi protozoa viruses

. mushroom
.
:
:

* ( )1212-1218
( ) .
* ) plaque (Yersinia pertusis 3/1

(25) . 111
. .
* ( ) 11 .

* .
:
-1 .

....DNA
.
] [
-2 .
21 .
( ).
-3 .

.
.

-4
bioremediation
.
-5
.
-6 .

-7 .. .

( .)1
.
3.8-3.5 .
11
] [

.
.2-11x 1521
.211 x
1676 ( .311x .)2
. .

"."animalcules
compound microscope bright
field microscope .
1111 x
(. )3
:
-1 .(10x) ocular lens
-2 objective lens (4x, 10x, 40x, 100x) .
x . condenser
.
resolving power
. .
.
0.2 micrometer
.
11
] [
oil immersion lens 100x
cedarwood oil

( . )4
*
:
*
.
* .
* Xylene
+.
* :
.
cotton swab .
12
] [

Isolation of bacteria from the environment

. .
- Hunging Drop Slide
* .
11 .
.
41.
:
*
* loop
* Hanging Drop Slide
* Swab
* nutrient broth nutrient agar
:
-1 .
-2 .
-3 ( .) 5
-4 11 . .
13
] [
- Norrmal skin flora

Normal flora commensals

.
(
) . :
Staphylococcus epidermidis -1
( )
Staphylococcus aureus -2
%21
( )
.
Micrococcus luteus -3
Propionbacterium acnes -4
( ).
propionbacterium granulosum -5 .
:
-1.
-2 .%71
.cotton swab -3
-4 1.2 %NaCI( saline ).
14
] [
-5 .nutrient agar
-1 %71
( (Transient organisms
-2 .
-3 swab 15.
-4
.
-5 24 37.
-6 ..

15
] [
PH
. .
loop
:
( )2-6
16
] [
Bacterial Staining

( ) .
saline
wet mount
* : :
- 1 Brownian movement
-2 :
flagella (.)11
- Smear Preparation
Smear
-1 .
-2
.
-3 loop
( )
17
] [
loop
loop .
(11-5) .
-4 3

-1 .
-2
.
-3 .
:SMEAR
( . )11
- (Positive and negative stains) Simple Stain
.
:
-1
( (chromophore ( (anion sodium+ eosinate - :
.
-2 ( )cation :
methylene blue+ chloride- .
NaOH .
18
] [
HCI
. (
) ( ).
( )
( ) .
Net charge
.
1-
Bacterial cell
Outer surface of most cells is negatively charged

+

()

)(Cation
()
)(anion
19
] [
Positive Stains Methylene blue
Crystal
. violet, basic fuchsin .

negative stains
.
.1 ( Bacillus subtilis ) 24.

.2 ( staphylococcus ) 24.
.3 methylene blue , crystal violet nigrosin
, .4 .
positive stain :
-1
: .
-2 .
-3 loop .
-4 loop .
.
. loop
.
.
-5 loop .

.
-6 (.
)12
-7
. .
21
] [
-
Negative stain :
-1
.
-2
loop
-3
: x11 x41
.
- () Differential stains
:
(mount
)wet
( . ).
.
.
-1 Gram Stain
1111 5111
.
Christian Gram 1883
:
-1 ( G+ ) ( )cocci
Staphylococcus aureus) (rods . Bacillus subtilis
21
] [
-2 ( G )
coli.
Escherichia
)Pseudomonas aeruginosa (rods) , (coccobacilli
.
:
-1 .
-2 crystal violet .
-3 .
-4 )mordant) Gram's iodine .

-5 .
-6 ( decolorizer )%25 .
.
-7 .
-8 ( counterstain) .
-2 .
( )14-13 :
-1 (.crystal violet (primary stain
(
) .
-2
( )crystal violet-iodine complexCV-I

( ).
-4 ( )

( . )15
22
-2
] [
)Schaeffer-fulton method( Spore staining
( endospore ).
sporulation
6-8 .

..
Bacillus--- rod-shaped-1 .
Clostridium rod-shaped - 2
Sporosarcina cocci,found in soil -3
Sporolactobacillus rod-shaped -4
Desulfolomaculum rod-shaped, ananerobic -5

Clostridium botulinum -1 botulism

.
Clostridium perfringens -2 Gas ganarene

Clostridium tetani -3 tetanus
Bacillus anthracis-4 .anthrax

.
* dipicolinic acid
dehydration
23
] [
.
. .
* .
thermostable enzymes
*
. :
-1 ()subterminal
-2 ()terminal
Clostridium tetani
( )drum-stick
. .
-3 (.)central
-1 ( 66-63 71.6 15
) .
-2

( .)16
-3 .

25 !
-4
.
24
] [
-1 Bacillus subtilis .
-2 ( aqueous5% .)%5
-3 .
-4 .
-5 .
( : )17
-1 .
-2 5
-3 .
-4 2.
-5 ( . )12-18
- 3 Acid Fast stain

Mycobacterium
( )Mycolic acids
.

.
.
Mycobacterium tuberculosis
Mycobacterium leprae
Tuberculosis
leprosy
25
] [
( )carbolfuchsin . ( )methylene blue .
( %3-1 %25 )
. ( ) .
:
( :)21

5 .
21-15.
31-21
: ( )21 leprosy
Nocardia .
-4 ( )
(Negative stain) Capsule stain
.Glycocalyx
capsule

Slime layer
dextran, glucan
26
] [
-1
Streptococcus mutans
( )Biofilm

-2 ( )phagocytosis
.
.
Streptococcus
pneumoniae .
microcapsule -3
81 Streptococcus pneumoniae
( . )23-22
( Capsule stain ) :
-1
.
-2
.
-3.
( Capsule stain )
-1 % 6 .
-2
.
-3 ""
-4
27
] [
-5 .
-6 .
-7 .
-8 .
-2 .
Hiss
( Capsule stain ) method
( .)24
-1
.
-2 .
-3 .
-4
20 %.
-5 .
:
* .
*
( .)25
28
] [
Flagella and motility

Flagellin .Flagella
basal body
.
.
( : .)26
-1 polar .
lophotrichous -2 .
peritrichous -3 .

Helicobacter Pylori
.
29
] [
* :
All spirilli
Half the bacilli
Few of the cocci
*
.
61
/ 5.2mm/Min
Thiospirillumaeruginosa
Pseudomonas
4.4mm/min Cheetah
111/ 25 ..
:
- ( Hanging Drop Method ).
- U
-:U-shaped tube U
.)%1.5
Stabbing Needle
loop 37 24 (
.)27
- -: Swarming
Proteus

( .)22-28
31
] [

Disinfection & Sterilization
.
.
..
* :sterilization (
.
.
* :disinfection .

( )disinfectants biocides

.germicides
.antiseptic
* :Decontamination
.
- physical methods :
- :Heat :
-1 :

Direct flaming
Incineration
-2 :
31
] [
) pasteurization
) Tyndalization
) boiling
) autoclave
- :radiation
Ionizing Radiation
-1
UV radiation
-2
Microwave
-3
- chemical methods
- mechanical methods
:
- :Heat
-1 :Dry Heat
6511-161 .
-1 Dehydration
-2
-3 Denaturation
()Powders
.
211 15
121 .
:
-1 loop Bunsen burner

1871 .
-2 Incineration
6511-811.
32
] [
-3 : 181-151 4-2

( ) .
.
.
-2 :Moist heat
.

. :
- :pasterurization
62
Mycobacterium tuberculos Salmonella typhi Brucella
. abortus pasteurization
flash
High
)Temperature Short Time (HTST 27
15 . % 22 27 .
21.111
/ .
3
Ultra High
Temperature 151-141

81 .
33
] [

saprophytes .
thermoduric
. thermophilic .
- :Tyndalization
. 111
61-31

. ( )
- :Boiling
. .
- ( :Autoclave ) 31
111

.
111
:
34
] [
-
Temperature
)(degrees centigrade steam pressure
POUND PER SQUARE INCH
111
118
115
11
121
15
126
21
.
( 121) .PSI 15
.
.
( )
.. .
- :Radiation
. .
:
-1 : X-rays
:
- ( )1.1-111.
- .
- .
( . ).
35
] [
. .

-OH superoxide -O .
.ethylene oxide .
carcinogen .
-2 :U-V radiation
221-231
( ) DNA RNA
U-V 261 .

.
.
Germicidal lamp
.
. U.V .
. U.V
. u.v .
36
] [
:Microwave -3

.
.
- :Chemical methods
-1 ( ) : ( )
%81-61
.

( ) .
%81-61
( ) .
-2 ( : )phenol carbolic acid

%5-2
. Lysol
. %12-5
.
-3 :Chlorine .
. Na-hypochlorite
. .
()Na-hypochlorite%5.25
1:111 ( )PPM511 .
.
* :
.
() 1:11 1:111
37
] [
( )Na or Ca - hypochlorite
-4 :Ozone ( )O3

.
-5:
formaldehyde glutaradehyde :)OPA) Orthophaladehyde

%2
glutaraldehyde
12-11
OPA . .glutaraldehyde
:formaldehyde -6 %37 .

:(Eto) Ethyleneoxide -7 .

Eto CO2
( ) 12-3 ().
38
] [
-1 .
11 3 7.8 3
.
-2
( )denaturation .cell membranes
-3 .
.
-4 glutaraldehyde
.
-5 Eto
.

-1
%85-61
-2 phenol
(
)
-3 (
).
-4 ()%3
-5 ( )
-6
denaturation

.

phospholipids

Neisseriae gonorrhoeae
39
] [

-1 (% 85-61 )

-2
-3

.denaturation

.
) )phospholipids

-4
( )
.
.
-5
.
(
-6
)
( %8-3
-7
)
.
-8( )%2

-2

-11

-11

-12

-13
denaturation
41
] [
- :Mechanical methods

cellulose acetate
cellulose nitrate .. .
chemically inert .
.micrometer 1.4-1.2
. ( .)31
( )High Efficiency Particulate Air HEPA
%22-27 1.3
.Micrometer
.
41
] [
Culture Media

Culture media
.
( C ) N ( )%21-71

" "Na,K,Ca,Mg ( )buffers

PH
PH .
KNaHPO4 + H2O
KH2PO4 + NaOH
) ( KNa2PO4 + H2O
KNaHPO4 + NaOH
) ( KH2PO4 + KCI
K2HPO4 + HCI
( ) .

. .
:
-1 Liquid Media:
nutrient broth, glucose broth, tryptic soy broth, brain
heart infusion :
-1 .
-2 .Fermentation
-3
. Biochemical tests
-4 thioglycolate broth
anaerobic and microaerophilic
sodium thioglycolate
.
42
] [
-2 Solid media
agar
:Agar polysaccharide
Geledium spp. :
-1
.
-2
(121(.
-3 28 45
.
-4 (.)%2-1.5
: 15
-1 .Gelatinase
-2 26 .
-3 ( )%15-11 ( )%25 -21

.
Nutrient Agar
Blood
Agar Chocolate Agar :

-1 .
-2 .
-3 .
-4 .
Petri Dish
- Deep Agar .
- Slant Agar
43
] [
-3 Semi Solid Media
SIM :
-1 :Synthetic Media
.
-2 :Non-Synthetic Media

Nutrient Broth Beet Extract Peptone

Yeast Extract .
.
:Peptone ( ) (
) ( ) .

.
.
44
] [
Preparation of Culture
Media

Magnetic stirrer .

PH adjustment
PH . : 5
"
PH"
1.1 HCl
1.1 NaOH 5
1111 3 :
( )
5
1.2
1111
= 41
PH PH-Meter PH - Paper
.
(121
15)
. 51 (
.) 32
45
] [

-1 :Enriched media
%11
%2
- :blood agar

51.
- chocolate agar:
25-21
.Haemophilus influenzae
- : Thayer-martin
Neisseria
.gonorrhoeae
-2 : selective media
.
-
.
- NaCL
Crystal
violet,antibiotics:
-1 Agar
MacConkey
bile salts
.
violet
crystal G+
G-.
46
] [
-2 Mannitol Salt Agar %7.5 NaCl
.Staphylococccus
-3 :Differential media

neutral red
Escherichia
coli
proteus mirabilis
( .)33
.EMB
MSA
Staphylococcus aureus

() ( .)34

Hemolysin
Streptococcus pneumoniae ( Alpha
)hemolysis .
Streptococcus pyogenes
.Beta hemolysis
Staphylococcus epidermidis
( gamma hemolysis .)35
47
] [
-4 special media:

( Lowenstein-jensen media )36

.
( . )37
:
:Pure Culture
Mixed Culture
.

-1 ( Spread Method:)38
:
-1 48-24 ( ).
- 2 () .%25
-3 . L
-4 . TSA
-5 .
:
48
] [
-1 .
-2 5-1 3
.
-3 .
.
-4 .
-5
.
-6 .
-2 ( :Streak Plate Method )39
- .
- (
) ( )1 Zigag
- ( )1 Zigzag
.
- ( )2 .
- 37.
-3 Pouring Plate Method (:)41

.
49
] [
and Anaerobic
Aerobic
Incubation

%21 ()
.

( : )41
( :Obligate aerobic bacteria )a (%21

) . Pseudomonas fluorescence .
.
catalase and
.superoxide dismutase
( :anaerobic bacteria )b
.
Clostridium botulinum
.
.
( :Facultative aerobic bacterria )c .

( )O2 ().
(ATP) . Escherichia coli

.
.
( :Microaerophilic bacteria )d (-2

)%11 .
Spirillum volutans Helicobacter pylori

.
superoxide
and
catalase
.dismutase
51
-
(bacteria )e
] [
Aerotolerant
. Lactobacillus
Streptococcus pyogenes .
.
: Deep agar -1
.
-2 :

.
-3 :
SH
...
:Anaerobic jar -4
vacuums
( ) .
(
) . )gas pack(
11
.
(
.)42
:Wright's tube -5 ()
pyrogallol .
51
] [
pyogallol 1 11 % NaOH
(. )43
:Brewers anaerobic petri dish -6

. Brewers anaerobic agar
thioglycolate acid (.
)44
-7 (
) ! oxygen
tesion .
-1 :
* Tryptic soy agar

.
* 45 . .
* .
( ) . .
37 24 .
-2 :
* Tryptic soy broth 5 .

.
* .
37 48 . ( )
( : + : ).
: - ().
52
] [

Quantitative measurements of bacteria growth

(1 )3
. .
: Direct Methods
Breed count -1 .
)counting chamber ) Haemocytometer -2 .

McFarland -3 .
Turbidity -4 .
: Indirect Methods
Plate count -1 .
Dry weight -2 .
Filter membranes -3 .
: Direct Methods
-1 ( Breed count .)45
-1 (1 ) 2 .
- 2 1.11 3
standard loop .
-3 . (
%25 ).
-4 Methylene Blue
(1 )3 :
= 11
16
53
] [
161 = 11 16
-1 = ( ) 2
= 2)81( 3.14
= 21126
1
-2 (1_________ = ) 2
21126 2
11 1111 (2 1)
21126 2
5111 4276 = 111.111.111 2
5111 1
5111 1.11 3
511.111 ( . )46
-2
)Counting chamber (Hemocytometer

) (Hemocytometer ( )47 .
. ( 1.1) .
. ()1/411
411 . (.)48
-:
-1
.
-2 .
-3 .
54
] [
-4
.
-5
. ( )1.1 3
. (.)42
: .
-1 .
* 111 = 511.
1/411
111 = 111 1/411

= 1/4
4 511
= 2111
= ( 1 1.1 )2
= 1.1
2111 1.1
3 ( )11
11 2111
= 21111 /
1111 1 = )mm( 3( 3)
1111
1 = 21111111/

:
55
] [
1 / 3 =
( )6114 ( ).
:
*

.
(. )51
: Indirect Methods
(Plate Count) Viable Count
. Plate Count
.

( : )51
.Spread Plate Method -1

.Pour Plate Method -2
Spread plate 11 3
.
.
Pour Plate (11 1.1)3
" ."8
311 31.
( ) 111/1 11/1
1.5 3 4.5 3 .
56
] [
:
3 . :
.
.
.
.
( )24
. .
: ( )311
.
(
) Colony Forming Unit
( )CFU .3
( )311-31
( )31 .
( )311 .
:
-1 Escherichia coli ( )24.
-2 ( )2 3 .
-3 .
-4 .
-5 ( 511 )3 .
57
] [
-1 ( )1 3 ( )2 3
. .
( )11/1 ( .)1-11
-2 ( )13 ( )1-11 ( )2 3
( .)2-11
( )6-11 .
-3 :
- :
-1 .
-2 .
-3 ( )1 3 ( )4-11
.
-4 .
. 37
( )48-24.
- : (:)52
-1 ( )1.1-1.2 3
.
-2 45.
-3 .
-4 " "8 .
-5 .
-6 .
-7 (-31
)311 . ( )31

PH .
58
] [
( )311
.)TNTC( Too Numerous To Count
-5 .
-6 :
1 / 3 = 11
( 1.1)3
Dilutions
-1 :
=
:- 1 - 1 3 2 3
:
()
= 2+ 1\1
= 11\1
= 11
1-
:-2 1.1 3 2.2 3

:
= 22 + 11\11
= 11\11
= 11
2-
59
] [
1 8.2 11 / .3
-
:
11
- 2
-
111 \ 1
11 \ 1 -
11-
3
5 \ 1 -
3 11\1 2.1 3 2.2 3

.
4 .
1.1 3 () .B, C, D
- A, B, C and D
- / 3
- 5 Escherichia coli 18 /2 11
61
] [

The Effect of environmemntial factors on bacterial growth
:
-1 :
.
-2
PH
C N,O
.
4:
-1 -4 .O2 -3 .PH -2 .
-1 :Effect of temperature

()
(111) .
Cardinal temperature
-1 .Minimum temperature
-2 . Optimum temperature
-3 . Maximum temperature
61
] [
().
( : )53
Mesophiles -1 .
51-11 41-21.
Escherichia coli 41-35

31 ( ) .
()
. 45
-2 (15 - 5- ) 15
psychrophiles
pseudomonas .

( Roquefort (blue cheese

( ) Penicillium roqueforti
-3 81 45
Thermophiles 51 - 45
Hyperthermophilic 113
Autoclave . .
.Thermostable
62
] [
* Refrigeration retards food spoilage becauseit limits growth of

mesophiles.
*
.
37 .
.
Hansens disease (leprosy) : (

) Mycobacterium leprae 31
( ) .
-2 ):Effect of (PH
(log -
)+1/H
. PH +H -OH
OH H+ .
PH ( )2-5 ( ( .)6-2 )54

PH
- 55
8-55
115-85
PH
1- Acidophiles
2- Neutrophiles
3- Alkalophiles
Shigella, Escherichia coli PH 2.5
511-11 Shigella
( )
pylori
Helicobacter PH
63
] [
-
NH4++ OH-
NH3 + H2O
PH .
.
.Buffers
Identification of Bacteria
). (
)
.
( )
.
( )Identification
( ) .
:
.1
(.)Pure Culture
.3
.2
( ).
.4
.6
.5
Enterobacteriaceae
64
[]
.
.
.Salmonella, Shigella :Pathogens ) (-1
Proteus,
:Occasional
Pathogens )( -2
.Klebsiella
Enterobacter,
Normal Intestinal flora )( -3

.Escherichia
:
Escherichia coli
Enterobacter aerogenes
Lactose fermenters
Salmonella typhimurium
enteric
Shigella dysenteriae
Proteus vulgaris
Lactose nonfermenters
Pseudomonas aeruginosa
Alcaligenes faecalis
Corynebacterium xerosis
Micrococcus luteus
Streptococcus lactis
Bacillus cereus
65
Nonenteric
] [
IMViC
Indole production test :I .

Methyl Red Test: M .
Voges Proskauer test :V .

For easier pronounciation :I .
Citrate utilization test :C .

-1 :Indole Production test
: ()Tryptophanase
: Pepton water :
( )
21.
5
PH= 74
)Semisolid Indol Motility( SIM
(.)Stabbing
37 24.
Kovacs Reagent : :

P- dimethylaminobenzaldehyde
151
11
51

P- Dimethylaminebenzaldehyde (
66
] [
) . Escherichia coli :
.Enterobacter aerogenes : (.)56( )55
-2 :Methyl Red Test
.
Escherichia coli. Enterobacter aerogenes
( )
) .
Lactic Acid
Formic Acid Acetic Acid .Succinic Acid

H2 , CO2 .Formic Acid
. Mixed Acid fermentation

( ) butandiol
fermentation (.)57
. Escherichia coli
PH 4.4 Enterobacter aerogenes
Acetoin (acetylmethylcarbinol) Ethanol
PH 6.
: Glucose phosphate
) MR VP medium ( peptone water 5/
% 11
51 ( 3 )
67
] [
7.6 =PH
37 28 - 24.
:Methyl Red :
11
311
211
PH ( )4.4 PH 6
( ) . .Enterobacter aerogenes
. Escherichia coli (.)58
-3 :Voges Proskauer
:
Butandiol
fermenters
butandiol, acetoin PH
Acetyl Methyl Carbinol
Acetoin Diacetyle .Butyelenglycol
MRVP medium Glucose phosphate peptone water
48.
Barritts reagent : O'meara reagent
alpha-naphthol
alchoholic %41 KOH
Acetyl Methyl Carbinol diacetyl

alpha-naphthol ( guanidine ) .
15 .
: . Enterobacter aerogenes
68
] [
Escherichia coli (.)52
:Citrate Utilization test -4
: ()
Citrate
permease
.
. PH (
) Bromothymol Blue .
Simmons Citrate Agar :
. IMViC
.
Bromothymol Blue Indicator
.Enterobacter aerogenes
: Escherichia coli
( )61 ( . )61 ( )62 . IMViC
69
] [
Sugar Fermentation

. .

-1 :Respiration ()
. .
-2 :Fermentation ( )

. .
- :)exoenzymes( :
.
- ( :)endoenzymes .
( )permeases
.
-1 Ex: Phenol Red Carbohydrate Broth

( )
( ) ( ).
71
] [
-2

.
-3 Durham Tube
. :
- .
- ( )63
(. )64
-4 24 37.
* (
)Durham Tube 121 15 .

.
.
71
] [
:)TSI(Triple Sugar Iron Agar

.
3 . (.)65
% 1.1 () %1 %1

. ( )Slant
. :
-1 : 3
.
PH 6.8
.
( )Butt

24 .

( ) .
-2 :

-3 :
H2S ( )
(.)66
FeS + SO3 + H2O
FeSO4 + H2 + S
Gelatinase
72
] [
() . Gelastinase
.

22 37 .
( )Proteolytic Enzyme Gelatinase

() .
:
-1
Staphylococcus aureus , Bacillus subtilis .
-2 ().
-1 ( ).
-2 35 48 24.
-3 5.
-4

( ) (.)67
73
] [
Oxidase Test
Oxidase
. Cytochrome Oxidase ( Cytochrome)

( )O2 H2o H2O2
Cytochrome Oxidase .
Microaerophilic
.Oxidase Pseudomonas Neisseria
Cytochrome Oxidease Cytochrome C
1% N, N, N, N tetramethyl-p-phenyldiamine ( :
)dihydrochloride .
( )
( ) Oxidase.

Tryptic Soy Agar
( )TSA 24
.
( )11-31 (.)68
74
] [

Phenylalanine Deamination
Phenylalanine Deaminase
.Phenylalanine
:
Proteus, Morganella,
.Providencia
:
CH2CHCOOH + NH3
Phenylpyruvic Acid
NH2
Phenylalanase
eeeeeeeee
CH2CHCOOH
Phenylaninie
Phenylaphyruvic Acid.
Phenylalnine agar :
37 24.
%11 Ferric Chloride :

:
)Ferric Chloride (anhydrous
Distilled water
11
111
11 - 5 .
.
5 1 (.) 62
75
] [
Nitrate Reduction Test
NO3
NO2 N2
.Reductases
Facultative Anaerobic
( ) .
.
Nitrate reductase
NO3 + 2e + 2H
NO2 + H2O
.
Pseudomonas .
N2
No3
No2
NO3
24 :
-1

:Reagent A
Sulfanilic Acid
-2
Acetic acid, 5m
:Reagent B
N, N- dimethyl - - naphthylamine
1.6
Acetic Acid 5m
211
211
5m acetic acid is prepared by adding 1 part of glacial acetic to 2.5

parts distilled water. Reagents A, B can be stored at 22c for 1 year.
. A,B 22
76
] [
Zinc Dust -3 : .
:
Nitrate broth :
-1 37 24-18 3
.A, B
-2 Zinc Dust
(.)1
-3 :
- = A, B
NO2
NO3
- A, B = zinc powder
NO2
NO3
N2
- zinc dust B, A = NO3
( zinc dust NO3, A, B )NO2
(.)71
77
] [
Urease enzyme

Klebsiella , Proteus
Helicobacter pylori
(
)
. CO2
.
:
Christensen's media
. .
121 15 .
(.)Slant
.PH
( )71
78
] [
:Catalase Test
( Staphylococcus )
Streptococci . Catalase
() :
2H2O + O2
Catalase
2H2O
:
-1 (.)Nutrient Agar
-2 .
-3 ( H2O2 %3 ).
: (
.) 72
:
Catalase .
79
] [
: Coagulase test

coagulase .
. Staphylococcus aureus
staphylothrombin
.
.
.Phagocytosis
Clumping Factor " "Slide Coagulase

.
.
.
11 3
153 . Citrated Human Or Rabbit Plasma

37 24 4
Clot (.)73
:
Blood = blood fluid (serum + plasma) + RBC + WBC + Platelets
(Plasma = blood - )RBC + WBC + Platelets
Serum =Blood (fibrinogen + cellular components).
-1
( )Anti-CoagulantK- Oxalate or sodium citrate or heparin :
12
3 1 3 %2
81
] [
. 1111
/ 15
.
Serum .
15 . .
81
] [

homopolysaccharide
D-Glucose
Amylase
)C6 H10 O5 )n C12H22O11

.C6H12 O6 .

. (. )74
amylopectin
)(bluish red or red brown
amylose
)(blue with I
Starch amylase
C6H10O5
)Amylopectin amylase dextrins amylase maltose (disaccharideMaltose

glucose
Colorless with iodine
amylase
-1
-2 48 37.
-3 . lugol iodine
-4 :

( )
.
82
] [

)Antibiotic Sensivity Test (Kirby- Bauer Method
:
:Antibiotic
( )Penicillium Cephalosporium Bacillus & Streptomyces

.
Paul Ehrlich
" "Magic bullet
606
( syphilis )Treponema pallidum salvarsan

( sulfanilamide ).
1228 Flemming
1241 .
.
:
* :Bacteriastatic
..
* :bactericidal .
* :Broad- spectrum G-
,G+
.

( ) .

( Clostridium difficile
.)pseudomembranes
83
] [
B-lactam
( )
-1 .
-2 .streptomycin, tetracycline
-3 Rifamycin .
: ()75
84
] [
Reductase test

- .
( )
.
35 6 .
( .)Methylene Blue Reduction Time MBRT

(6) .
- 35 .
.
Escherichia coli
Streptococcus lactis ,
: ()76
-1 13 ( ) 1:25111
11 3 .
-2 3 .
-3 35 . 6
.
. .
85
] [
Evaluation of disinfectants
A-Phenol coefficient
B- Filter paper method
C- use- dilution method
1811 .
11111 1111
.
-1 .
-2 .
-3
.
-4 .
-5 .
-6 .
-7 .
-1 )%37( High level :

()

-2 :Intermediate
.
-3 :Weak Level
.
86
] [
: phenol coefficient -1
.
-2 : .

.
-3 :Use dilution method
.
.

. quaternary detergents
Use dilution method
.:
-1 Staphylococcus aureus
-2 ()
2\1
.
-3 ( )
( )
.
-4 ( )
( ) . (
).
-5 1 .
-6 .
-7 Staphylococcus aureus
( ).
-8 48-24 37.
.
( )77
87
] [

Bacterial Identification Using Rapid MuIti-test CommerciaI Systems
Enterotube II API 20E minitek
( )Pattern
( )
() (.) biotype
cupuIe
()

. 20 .
:
-1 .
-2 .
-3 .
-4.
-5 .
.

API 20E :
88
] [
API 20E 20
( ) .
Oxidase
.21
-1 () .
-2 5 (
).
-3 .
.
-4 SaIine ( 0.85% NaCI ) or Suspension media 5.
-5
cupuIe
.

37 48-24
:
-1 IND .
-2 A and B to VP 10.
-3 .FeCI3 to TDA
-4 .
89
] [
-5 ( 1 2 4 0
).
-6 ( + ) -
( phenyIaIanine VP and ).
-7 .
.
-8
3-2.
-2 ( + ) -
.
-11 7 .

(.)78
91
[]
References
Foundations in Microbiology, by Kathleen Talaro and Arthur

Talaro. 1996.
Fundamentals of Microbiology. Martin Forbisher, ninth edition.
1974.
Laboratory Exercices in Microbiology, John P. harley, and
Lancing M. Prescott. Third edition. 1996.
Microbiological Applications, Lab manual in general
microbiology. Harold Benson. eighth edition. 2002.
Microbiological experiments: A Health science Perspective,
Third edition.
Microbiology , a human perspective. Eugene W. Nester, 1995.
Microbiology. Lab Exercises. Margaret Bamett, Second edition.
1997.
Microbiology: A Laboratory Manual. James G. cappocino. 1987.
Microbiology, Dynamics and diversity. Jerome J. Perry, 1997.
Microbiology, Lancing M. Prescott. 1990.
Principles of microbiology, by Ronald M. Atlas. First edition,
1995.
91
( )
[
( )
( )
( )
:( )
( )
( )
[
( )
( )
)
[
:(
:(
:(
:(
)
[
:(
. Bacillus cereus
:(
:(
Clostridium tetani
)
[
:(
:(
:(
:(
)
[
:(
:(
)
[
:(
:(
:(
)
[
Escherichia coli
Proteus mirabilis
:(
:(
)
[
Streptococcus pyogenes
Enterococcus faecalis
Streptococcus pneumoniae
:(
( )
(lowenstein - jensen media)
:(
)
[
Streaking.
:(
:(
:(
:(
Brewer
:(
. Micrometer
.(
:(
)
[
:(
:(
.
.
.(
:(
:(
)
[
:(
:(
:(
:(
:(
)
[
.MR , VP
:(
:(
:(
:(
)
[
:(
.IMViC
:(
:(
:(
TSI
:(
)
[
.TSI
:(
:(
:(
:(
)
[
:(
)
[
:(
)
[
:(
)
[
.(
:(
Bacillus subtilis)
:(
:(
)
[
Pseudomonas aeruginosa
API 20E
)
[

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] [

. 12 ............................ Hanging drop Slide

. 13 ......................... Normal Skin Flora

. 17 .................................... Simple Stain

. 22 ............................ U-Shaped tube

- ( 82 ...... )Kirby Bauer

- 84 ..................................... Reductase Test

()lowenstein - jensen media

API 20E Pseudomonas aeruginosa

fungi protozoa viruses

* ) plaque (Yersinia pertusis 3/1

1676 ( .311x .)2

compound microscope bright

-1 .(10x) ocular lens

-2 objective lens (4x, 10x, 40x, 100x) .

oil immersion lens 100x

- Hunging Drop Slide

* Hanging Drop Slide

* nutrient broth nutrient agar

- Norrmal skin flora

Normal flora commensals

-4 1.2 %NaCI( saline ).

- (Positive and negative stains) Simple Stain

Positive Stains Methylene blue

. violet, basic fuchsin .

.1 ( Bacillus subtilis ) 24.

.3 methylene blue , crystal violet nigrosin

)Pseudomonas aeruginosa (rods) , (coccobacilli

-4 )mordant) Gram's iodine .

-1 (.crystal violet (primary stain

)Schaeffer-fulton method( Spore staining

Sporosarcina cocci,found in soil -3

Desulfolomaculum rod-shaped, ananerobic -5

Clostridium botulinum -1 botulism

Clostridium perfringens -2 Gas ganarene

Bacillus anthracis-4 .anthrax

( )carbolfuchsin . ( )methylene blue .

Flagella and motility

- ( Hanging Drop Method ).

-1 loop Bunsen burner

Mycobacterium tuberculos Salmonella typhi Brucella

)Temperature Short Time (HTST 27

POUND PER SQUARE INCH

-2 ( : )phenol carbolic acid

formaldehyde glutaradehyde :)OPA) Orthophaladehyde

( )High Efficiency Particulate Air HEPA

" "Na,K,Ca,Mg ( )buffers

-3 ( )%15-11 ( )%25 -21

Agar Chocolate Agar :

-3 Semi Solid Media

Nutrient Broth Beet Extract Peptone

-2 Mannitol Salt Agar %7.5 NaCl

Streptococcus pneumoniae ( Alpha

( gamma hemolysis .)35

( Lowenstein-jensen media )36

-2 ( :Streak Plate Method )39

-3 Pouring Plate Method (:)41

( :Obligate aerobic bacteria )a (%21

( :Facultative aerobic bacterria )c .

(ATP) . Escherichia coli

( :Microaerophilic bacteria )d (-2

Spirillum volutans Helicobacter pylori

:Brewers anaerobic petri dish -6

. Brewers anaerobic agar