(Workbook for General Microbiology)

(Workbook for General Microbiology)

:
.
.

2558


(Workbook for General Microbiology)
700201
710 231
(Workbook for General Microbiology)
710 231

standard plate count
streak plate
Disinfectant, antiseptic antibiotic Selective
Differential Media, Extracellular Intracellular enzyme


2 Unknown

2558

1
2 Pour plate
3
4
5
6 Disinfectant Antiseptic
Antibiotic
7 Selective Differential media

8 Extracellular Intracellular enzymes

9
10
11 Unknown
12

______________________

2
4
5
11
20
24
28
33
39
43
48
52
54
57
60

(safety first)


1. () ()
()

2.
3.
4. ( 70 )

5. (aseptic technique)
(sterilized)
6.
7.


8.
9.

10. , ,
11.

12.

13.

(Code

number)


14.
(autoclave) (water bath) (hot air oven) (Incubator)
(colony counter)
(
)
15.

16.

17.

18.

1. Plate (Petri dish)

2. 200 .
3. Beaker Flask 500 .
4. (Test tube) 15 .
5. Rack
6. (stirring rod)
7. Pipette 10 .
8. Pipette 1 .
9. Spreader
10. Glass funnel
11. Slide
12. Loop
13. Needle
14. Dropper + Dropper bulb
15. Wash bottle
16. Spray bottle
17. Alcohol lamp
18. Pipette bulb + tip
19. Eppendorf
20.
21.
22. label
23.
24.
25.
26.
27.

1

(Basic Apparatus in Microbiological
Laboratory)

1.
(Inoculating loop and needle)
2.
Alcohol / (Bunsen burner)
3.
(Petri dish)
4.
(Culture tube)
5.
(Pipete)
6.
(Staining set)
7.
(Incubator)
8.
(Refrigerator)
9.
(Water bath)
10. (Autoclave)

1. (Inoculating loop and needle)


4

2. (Bunsen burner)
,
, ,
3.



4. (Petri dish)
2
(agar) (surface area)

5. (Culture tube)
(Test tube)
(screw cap test tube)

(agar) (broth)

6. (Pipette)

7. (Staining set)
Grams stain, Acid fast stain
Simple stain

8. (Incubator)

37 C

9. (Refrigerator)

, , ,

(
)
10. (Water bath)

11. (Autoclave)


(Chamber)
121C
15 / 10-15

(sterile) (media)
,
13. (Hot air oven)
(Dry heat) 180C 2

1

1.
(Inoculating loop and needle)

2.
Alcohol / (Bunsen burner)

3.
(Petri dish)

10

4.
(Culture tube)

5.
(Pipete)

6.
(Staining set)

7.
(Incubator)

11

8.
(Refrigerator)

9.
(Water bath)

10. (Autoclave)

11. (Hot air oven)

12

2

Pour plate
2
1. (physic method) (total or direct count)
(turbidity)
2. (biological method) plate count, drop
count, surface count, membrane filter most probable number
Dilution plate count standard plate count

1.
2. Dilution plate count
3. (dilution) 10-fold dilution

1:

1
1. Nutrient broth
2. Agar
3. 200 .
4. Water bath 65
5.
6.
7. Hot plate

13

2
1. Beaker Flask 500 . 1
2.
3.
4. 200 . 3
5. label

1. NB beaker
flask 500 .
2. 200 .
3. Agar 1.5% 2
4. Hot plate ()
5. 2 3

autoclave ()
6. autoclave water bath 65

1: (Sterile Distilled Water)

1
1.
2
1. Test tube 6
2. Rack 1
3. Beaker flask 500 . 1
4. Pipette 10 . 1

14

1. Beaker flask 100 .

2. 6 rack
3. Pipette 10 . 9 .
4. ()
autoclave ( )
5. (Autoclave)

1: Sterile Pipette

1. Pipette 1 .

1. Pipette 1 .
2. (Autoclave)

2: Pour plate

1
1.
2
1. Sterilize 9 . 6
2. Sterilized pipette 1 . 7
3. Sterilized petri dish 2 1

15

1.
2. Serial dilution 1 . 1 .
9 . 10 1: 10
10-1
3. 2 1 .
9 . 10 1: 100
10-2
4. 3 1 .
9 . 10 1: 1,000
10-3
5. Serial dilution 1:1,000,000 10-6
6. 1 . 2 10-5 - 10-6
0.1 . Sterilized Petri dish 1 1
Nutrient agar
7. Incubator 24
30 300

3:

Standard plate count (SPC)

Aerobic plate count (APC)
Standard plate count (serially
diluted) (aliquotes) (dilution)
(non-selective medium)

(Cfu/ml) = X
Petri dish

16

10

*** plate 30 300
4 ***

2.1
1 .

1 .

10-1

9 .

10-2

1 .

10-3

1 .

10-4

1 .

10-5

1 .

10-6

17

2.2 pour plate spread plate

: T.D. Brock (2000)

18

2.3 plate

:
http://www.sciencebuddies.org/mentoring/project_ideas/MicroBio_Interpreting_Plates.shtml

19

2
() ................................................
..................................................................... CFU/ml
( Form, Elevation, Margin, Surface, Opacity
( ) Pigment ( ) 2.3 )
..............................................................................................................................................................................
..............................................................................................................................................................................
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20

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21

Steak plate method, Loop dilution technique, Heat

shock, Differential plating, Selective plating
Steak plate method
cross steak

Steak plate method

1:

1
1. Nutrient broth
2. Agar
3. 200 .
4. Water bath 65
5.
6.
7. Hot plate

22

2
1. Beaker Flask 500 . 1
2.
3.
4. 200 . 3
5. label

(Autoclave)
water bath 65

1: Sterile Plate

1. Petri dish 10

1. Petri dish 5
2. (Autoclave)

2: Plate Steak plate

1. Sterile Petri dish 10

2. NA 65
3. Loop
4.

23

1.
2. NA 65 plate sterile
plate 15 20 plate
plate
3.
4. plate
5. loop

4-5 (
) 2-3
streak

6. plate steak Incubator 37
18 - 24
3.1 Steak plate

3: steak plate

*** plate
4 ***

24

3
( Form, Elevation, Margin, Surface, Opacity
( ) Pigment ( ) 2.3 )
..............................................................................................................................................................................
..............................................................................................................................................................................
..............................................................................................................................................................................
..............................................................................................................................................................................
..............................................................................................................................................................................
..............................................................................................................................................................................
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25

1:
1
1. NB
2. Agar
3. Hot plate
2
1.
2.
3. microcentrifuge tube eppendorf tube

1. NA
2. 2 (Autoclave)
(Slant)
3. NB 1 (Autoclave)

4. NA semisolid 1 ampoule semisolid

NA agar 0.5%
(Autoclave)

26

2:

1. plate
2.
60

3.


4.
4.
5. label
6. loop

incubate 24
7. loop stab
1 incubate 24
3:

***
4 ***

27

4.1 NA

NA
Slant

NA Slant

Slant

28

4
( Form, Elevation, Margin, Surface,
Opacity ( ) Pigment ( ) 2.3
/ )
..............................................................................................................................................................................
..............................................................................................................................................................................
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29

5



2
1. (negative)
india ink negrosin
2. (positive)
simple strain
differential strain
2
differential staining 2
primary strain cystal violet counter strain
secondary strain safranin o
mordant crystal violet
(decolorization) (alcohol) (acetone alcohol)

(Lipid)
crystal violet iodine

1.
2.

30

1: Slide
1
1.
2.
2
1. Slide
2.

1.
2. permanent slide
3. loop loop
slide
4. crystal violet 1
5.
1
6. 95%
20
7. safranin O 1
8.
9.
***:

31

2 3:

5.1

5.2

: http://tutorials.jakearkinstall.com/article/view/4

32

5.3

: http://tutorials.jakearkinstall.com/article/view/4

33

5
( Shape, , )
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34

6
Disinfectant Antiseptic
Antibiotic
Disinfectant
Antiseptic Disinfectant
antiseptic
Antibiotic
Antibiotic S.A.Waksman .. 1945
penicillin ( . 1929)

1. spread plate
2. Agar diffusion method
1:

1
1. Nutrient broth
2. Agar
3. 200 .
4. Water bath 65
5.
6.
7. Hot plate

35

2
1. Beaker Flask 500 . 1
2.
3.
4. 200 . 3
5. label

NA (Autoclave) water bath

65

1: Sterile Plate

2
1. Petri dish 10

1. Petri dish 5
2. (Autoclave)
1:

2
1.

1.
2. (Autoclave)

36

1: pipette sterile

2
1. Pipette 1 .

1. Pipette 1 .
2. (Autoclave)
2: plate spread plate

1
1. NB broth
2. antibiotic disc
3. antiseptic ethanol dettol
2
1. Sterile Petri dish 10
2. NA
65
3. loop
4.

1.
2. NA 65 plate sterile
plate 15 20
3. permanent marker NA 4 C
control antiseptic 3
4. plate plate 2 antibiotic

37

5. Pipette 0.1 .
spreader ( )
6. sterile C
7. antiseptic ethanol dettol

8. antibiotic disc
9. plate
Incubator 24

3:

Antiseptic antibiotic

Disk method Agar diffusion method filter paper

method

(Clear zone)

***
***

38

6.1 4 antiseptic
antibiotic

39

6
1. Clear Zone Antiseptic
a. 1 ..................................... .............
b. 1 ..................................... .............
c. 1 ..................................... .............
d. 1 ..................................... .............
2. Clear zone Antibiotic
a. 1 ..................................... .............
b. 1 ..................................... .............
c. 1 ..................................... .............
d. 1 ..................................... .............
3. Antiseptic Antibiotic
......................................................................................................................

40

7
Selective Differential media



Catabolism Anabolism

Selective media

Differential media metabolism

Selective Differential media

1:

1
1. MacConkey agar
2. Mannitol salt agar
3. Salmonella-shigella agar

41

4. MRS agar
5. Eosin Methylene blue agar
6. 200 .
7. Water bath 65
8.
9.
10. Hot plate
2
1. Beaker Flask 500 . 1
2.
3.
4. 200 . 3
5. label

1. 5 100 . beaker
flask 500 .
2. Hot plate ()
3. 2 3

(Autoclave) ( )
4. (Autoclave) water bath
65

42

1: Sterile Petri dish

2
1. Petri dish 10

1.
2. (Autoclave)

2: Plate Steak plate

2
1. Sterile Petri dish 10
2. 5 65
3. loop
4.

1.
2. 5 65 plate sterile
plate 15 20 2 plate
3. plate
4. loop steak plate
5. Plate steak Incubator 37
18 - 24

43

3: steak plate

44

7
1.
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2.
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.. ..........................................................................................................................................................................
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.... ........................................................................................................................................................................
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...... ......................................................................................................................................................................
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45

8
Extracellular Intracellular enzyme

Metabolism
Extracellular enzyme

Intracellular enzyme enzyme

Extracellular enzyme Catalase Intracellular
enzyme

Metabolism
1:

1
1. Nutrient Broth
2. Agar
3. Soluble starch
4. U.H.T. milk (low fat)
5. Calcium Chloride (CaCl2.2H2O)
6. Tween 80
7. Peptone
8. Sodium Chloride
9. 200 .

46

10. Water bath 65

11.
12.
13. Hot plate
2
1. Beaker Flask 500 . 1
2.
3.
4. 200 . 3
5. label

1.
. Starch agar
1
Soluble starch
20 g NA 1
. Lipase test medium
1
Peptone
10 g
Sodium Chloride
5g
Calcium Chloride
0.1 g
Tween 80
10 ml
Agar
15 g

1000 . (Autoclave)
. NA+ milk
1
NA
1
U.H.T milk (low fat) 250 ml.
Agar
4.5 g

47

Distrilled water

150 ml.

1) NA (Autoclave)
2) 4.5 g 150 . (Autoclave)
3) NA sterile
4) U.H.T milk 250 . NA 8
10 .

1: Sterile Petri dish

2
1. Petri dish 10

1.
2. (Autoclave)
2: point inoculate

2
1. Sterile Petri dish 10
2. 3 65
3. loop
4.

1.
2. 3 plate point inoculate

48

3. Plate point Incubator 37

18 - 24
3:

Iodine point

(Precipitation zone)
zone
Clear zone
zone
Catalase
Catalase Hygragen peroxide (H2O2)
3 10% Hydrogen peroxide
catalase (catalase+) catalase (Catalase-)

49

8
..................................................................................................................................................
................................................................................................................................................
.............................................................................................................................................
catalase..

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50

9

(Lactic
acid) (Probiotic)

1:

1
1. BHI
2. MRS agar
3. 200 .
4. Water bath 65
5.
6.
7. Hot plate
2
1. Beaker Flask 500 . 1
2.
3.
4. 200 . 3
5. label

51

1.
2.
3.
4.

BHI Soft agar ( agar 0.7%) 10 .

BHI Broth 5 .
MRS Agar 200 .
(Autoclave)

1: Sterile Pipette
2
1. Pipette 1 .

1. Pipette 1 .
2. (Autoclave)

1: Sterile Petri dish

2
1. Petri dish 10

1.
2. (Autoclave)

2: Lactic

1
1. Lactobacillus spp. MRS broth 1

52

2
1. Loop
2.
3. label

1. BHI 3
Incubator
2. MRS agar
3. Lactic MRS spot 4
4. sterile pipette BHI 3 BHI Soft agar 0.7%
40 loop spot
5. incubator 24

3:

53

9

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54

10

Superkingdom Eukaryota Kingdom Fungi Myceteae
(filamentous) Hypha

1-2-3:

1
1.
2.
2
1. Slide
2. Cover slip
3.

2 3
cover slip
40

55

10

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56

11
Unknown



1.

2.
3. Intracellular Extracellular enzyme

700201

1. Unknown
2. 2
3. 1
1
4. Unknown
5. Unknown

57

11

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58

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59

12

1.

2. 700201

1.
2. 2
3. 1
1
4. E.coli Salmonella spp.

5.

60

12

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61

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62

. 2547. 2.
.
. 2547. . .
. . . .
Phrutiya Nilpratruck and Mana Kanjanamaneesathian. 2549. Laboratory Manual Microbiology
for Agricultural Students. Faculty of Animal Science and Agricultural Technology,
Silpakorn University.

63

:
E-mail: makemerry@gmail.com
: (2543)
(2545)
() (2552)
Doctor of Philosophy (Pharmacy), Chiang Mai University (2552)
:
:
1. Kasornpikul Charunee, Chaiyavat Chaiyasut, Bussabun Sirithanyalug, Worapol Aegwanich and Thanit
Pewnim. Physiological and Biochemical Characterization of Lactic Acid Bacteria Isolated from Thai
Native Chicken Faeces for Using as Probiotics.(1st Silpakorn reseach fair, 2007)
2. . 2551. .

3. Kasornpikul Charunee, Chaiyavat Chaiyasut, Bussabun Sirithanyalug, Worapol Aegwanich and Thanit
Pewnim. 2008. Probiotics Properties of Bacteria Isolated from Poultry in Thailand. Poster presented at
The International Society for Southeast Asian Agricultural Sciences (ISSAAS) Internation conference.
Bangkok. December 15 19.
4. Korpraditsakul Vichai, Charunee Kasornpikul, Surawat Chalorsantisakul. 2008. Effect of Tree Onion
extracts as prebiotics on Newcastle disease vaccine titer in Broiler. Poster presented at The
International Society for Southeast Asian Agricultural Sciences (ISSAAS) Internation conference. Bangkok.
December 15 19.
5. Kasornpikul Charunee, Chaiyavat Chaiyasut, Bussabun Sirithanyalug, Worapol Aegwanich and Thanit
Pewnim. 2008. Lactobacillus plantalum CMU-FP002 induction local protective immunity to Eimeria
tenella. Poster presented at The second International Conference on Natural products for Health and
Beauty. Phayao. December 17 19.
6. Charunee Kasornpikula*, Chaiyavat Chaiyasuta, Bussabun Sirithanyaluga, Worapol Aeagwanichb and
Thanit Pewnim 2009. Effect of the probiotic Lactobacillus plantalum CMU-FP002 on oocyst shedding
by broilers inoculated with Eimeria tenella. Avian Biology Research 2 (3), 157159.

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Doctor of Philosophy (Pharmacy), Chiang Mai University (2554)
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6. Korpraditsakul Vichai, Charunee Kasornpikul, Surawat Chalorsantisakul. 2008. Effect of Tree Onion
extracts as prebiotics on Newcastle disease vaccine titer in Broiler. Poster presented at The
International Society for Southeast Asian Agricultural Sciences (ISSAAS) International conference.
Bangkok. December 15 19.

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