14.6.

2011

'

.,.

" :
, ' ,
.2011

038093431

 200495034
:

-2

,

'

 : 038093431
200495034
:

:
.........................................................................................................................
1 .............................

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3 ..................
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5............................

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15.......

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18...

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19...............
......................................................................................................................
20............................

 ' -1
...................................................................................................................
20
' -2
20...........................................................................................................
' -3 "
'21......................................................
' -4 Plaque
assay.............................................................21
' -5 A
22 .........................................................................
' -6 B
22 ........................................................................
' -7 C
23 ........................................................................
' 8 A1, B1,
C1................................................................................24
' 9 A
6 ...................................................................
' -10 A
25.................
' 11 B
7 .................................................................
' -12 B
25 .................
' 13 C
8 .................................................................

 ' -14 C
25 ................
' -15 A
'26.....................................................................
' -16 B
'26.....................................................................
' -17 A....................................................................
.........................26
' -18 B...................................................................
.........................27
-19
'14 .......................................................

-1 A
5 ..............................................................................
-2 B
5 ..............................................................................
-3 C
6 ..............................................................................
-4 A...........................................................................
..........................7
-5 B..........................................................................
...........................8
-6 C..........................................................................
...........................9

 -7 A"
'10..............................................................
-8 B"
'10..............................................................
-9 A '
11.................................................................
-10 A '
12.................................................................
-11 B '
12................................................................
-12 B '
13..................................................................
-13 B '
13..................................................................

:
, . ,
. )
(generation ,
. - ,PH , ,
).(A1
.Lag ,
-phase . :
,
, .V0Exponential ,
-phase .
- , , ' ,
. -
, PH , , .
:
.(k=0.301/g (1.1 .
N=N02 :
,(n (1.2 N N0 , n ,
. -n :
.(n=3.3(logN-logN0) (1.3 :
,(g=t/n (1.4 -t . -Stationary phase ,
,
." :
, .
. -Death phase ,
, .
, ).(B1
) ( ) (.
: ,pH , ,,
. ,
, ,/ , .

: , -
, , . ) ,(c0-40 )c
,(37-390 ) ,( c600 ) .( c80-1100 -PH ,
PH . : - -
PH ," PH=6- - PH-
," .PH=9- , - .
) (nonhalophile ,
) (Halotolerant
, - ) (
- ).(C1
, .

, .
.
.
) .(capsomer
.nucleocapsid "
).(D1
' . ,'
.
. , -
.
, ' . , -
. "
,
. , -
- , .
. , -
)( . ,-
.
one step growth curve :
- ' ) (,

. .
- ,
.burst size - '
. ' ' .
, '
.
,"
. " .-

(E,21

.(plaques assay

:
.
.
' .
" '.

:
:
LB

LB pH
LB

'

-
pH ,
) .(A,B,C pH
) 1 ,(23
. ,
. , -
( ( .

CFU
, ((colony forming units .

) .(F1 A,1B,1C1
) 2 .(23 , -
. ,
.
)
( . , ,
) .(G1
) (1 8.
" ' -
A .B- ' ) 3
.(24 ' , 37OC -
10 . '
, . .LB
. 37
OC 40 .
-Plaque assay .
. ' ) (
) .(plaque ,
' ' )
(
. ' ,
' .
100% ,
) ( . ,
(3

) .(PFU (plaque forming units A B-

5 , 24- ) 4 (24

' - A,BC-
' , - . ' ,
.

:

A:

-1 A . ' ) 5
.(25
A1 ,A5 -

pH -

10.0 . pH= 7.0 ,A4 A6 .A7

:B
-2 B . ' ) 6
.(25
,B1 B3, B5-
,B6 .
A4 .A7
:C
-3 C . ' ) 7
.(27
,C6 C3, C5-
,C7 .
C2 .C4

) 1
(:
' 9 A :

1.93787E+09
1.585E+09
1.646E+09
9.613E+08
0.000E+00

3.275E+08
3.710E+08
7.060E+08
6.950E+08
0.000E+00

)(
0
120
240
360
480

1A :
=3.275E+080.169=1.93787E+09 bacteria1 OD F=Number of bacteria for mlOD at the
same time point

:A

-4 .A ) 10 (27
log, lag - ,stationary - log -
.
1.3:1.4-
=t3.3*(logN-logN0)=300-1803.3*[log(9.858E+08)-log(4.246E+08)]=99.4 min g=tn

' 11 B :

2.21656E+0
9
1.463E+10
9.600E+09
1.183E+10
0.000E+00

)(

3.48E+08

3.60E+09
4.56E+09

120
240
360
480

7.915E+09
0.000E+00

:B

-5 .B ) 12 (28
log, lag - ,stationary - log -
.
1.3:1.4-
=t3.3*(logN-logN0)=20.6 min g=tn

' 13 C :

6.62651E+0
7
3.203E+08
4.840E+09
1.327E+08
0.000E+00

0
1.650E+07
120 1.105E+08
240 6.205E+09
360 3.650E+08
480 0.000E+00

:C

-6 .C ) 14 (28
log, lag - ,stationary - log -
.
1.3:1.4-
=t3.3*(logN-logN0)=15.8 min g=tn

" ':

-7 A" ' . ' 15

) (
A ' ' ,
) 1A ' ( 180- , ) ' .(

-8 B" ' . ' 16

) (

B ' '
) B2 ' ( 300- , ) ' .(
:burst size
:A

-9 A ' . ' 17
) (.
:burst size
Burst size=3.580E+103.540E+08=101.13

Burst time= 300 min= 5 hr. ,

PFUml*min

Burst size=5.700E+103.580E+10=1.6

Burst time= 1140 min= 19 hr. ,

PFUml*min

-10 A ' . ' 17

) (.
:burst size
Burst size=4.720E+091.100E+09=4.3 PFUml*min

Burst time= 300 min= 5 hr. ,

:B

-11 B ' . ' 18

) ( . burst size .

-12 B ' . ' 18

) (.

:burst size
Burst size=4.213E+093.7E+07=113.9 PFUml*min

Burst time= 300 min= 5 hr. ,

-13 B ' . ' 18

) (.
:burst size
Burst size=2.640E+114.213E+09=62.7

Burst time= 1140 min= 19 hr. ,

PFUml*min

-19 ':
B

+'

-'

A ' , B '
C ' .
.

1 A 1
-5 37OC ,NaCl 0.5% pH 10.0 -
.pH= 7.0 ,7 6 ,4
A -pH .0.5%
B ,2 37OC, pH -1
7.0 .NaCl 0.5% - pH , )(0.05%
,OC45 ,
B .
7 ,4- pH - ).(5%
3 ,C -6
37OC pH 7.0 ) .(0.05% ,
) ,(0.5%/5% OC45 ,pH=10 -
, C . ,
pH
.
1.5-
, .
,C 15.8
. B 33.9 .
A 99.4.
-OD,
) .(1 OD-
,
. ,A lag-
.

, .
, OD- .

,log- ,
. A
. ,

.OD- stationary-
, . ,C
lag- ,log- death-
, . B
lag- .log- log-
,
.A A lag
B ,C- A .
,6 - 4 . C
) 15.8 -( . -
,stationary , A C-
, . C
A ,log- stationary -
C .A ,
. " ,

C .A
stationary - ,death -
)( 4 .6-
stationary .B
, B
. / OD
. , B
) .(log -
'
' . ,
' , A ' , B '
. ,.

" '
' , ,
. ' ,
,
, . ,7 A '
' . ,
180 - ) A1 ' (
) ' ,( '
. Plaque assay
,
' . A1
" .burst size . A2 '
0 5 - , 19 .
, A ' . -
0 ,5- . ,
,A2
A1 . ,8 B '
, . 300
) B2 ' ( ) B1 ' ,(
' . B2
" .burst size . B1

0 19- , ' .
, 5 .
.A2

:
Bacteriophages wastewater treatment :
http://www.scialert.net/fulltext/?doi=jm.2007.889.899&org=10
Bacteriophages Isolation
http://www.dairyscience.info/isolation-and-purification-of-bacteriophages.html
http://www.bio.davidson.edu/people/dawessner/302/302Lab6.html
http://phagetherapylightandshade.blogspot.com/2009/04/sources-andbacteriophages-isolation.html

162-168 -

Madigan M. , Martinko J. , Dunlap P. & Clark D. (2009), Brook biology of.1

microorganisms. (12nd Ed), San Francisco, Pages: A- 142; B- 147-150; C.157-168; D- 252-256; E- 258-270,

F-153-155, G-156-157
Laboratory Brochure.2

Plaque assay is taken from:.3

http://www.virology.ws/2009/07/06/detecting-viruses-the-plaque-assay
.", using search word "Plaque assay

:
: -1 '

1
2
3
4
5
6
7

37OC/ pH 7.0 / 0.5% NaCl
RT / pH 7.0 / 0.5% NaCl
45OC / pH 7.0 /0.5% NaCl
37OC / pH 2.0 / 0.5% NaCl
37OC / pH 10.0 / 0.5% NaCl
37OC / pH 7.0 / 0.05% NaCl
37OC / pH 7.0 / 5% NaCl

: -2 '

10-5

10-6

10-7

A1

0h

A1

2h

A1

4h

A1

6h

A1

8h

B1

0h

B1

2h

B1

4h

B1

6h

B1

8h

C1

0h

C1

2h

C1

4h

C1

6h

C1

8h

' -3
2A

1A

2B

1B

" ':

' -4 :Plaque assay

10-8

10-7

10-6

10-5

0h

5h

24h

1A or B
2A or B
1A or B
2A or B
1A or B
2A or B

' -5 A :
7A

6A

5A

OD
4A

3A

2A

1A

37OC

37 C

37 C

37 C

45 C

RT

37 C

pH 7.0

pH 7.0

pH 10.0

pH 2.0

pH 7.0

pH 7.0

pH 7.0

NaCl 5%

Na 0.05%

NaCl 0.5%

NaC 0.5%

NaCl 0.5%

0.5% NaCl

0.5%

Cl

0.265
0.385
0.421
0.415
0.403
0.503
0.531
0.549
0.421

0.205
0.341
0.348
0.239
0.282
0.350
0.294
0.350
0.208

0.209
0.364
0.430
0.531
0.604
0.826
1.042
0.444
0.533

NaCl

0.276
0.322
0.329
0.322
0.268
0.359
0.389
0.359
0.233

0.150
0.227
0.267
0.273
0.435
0.496
0.574
0.543
0.477

0.179
0.187
0.184
0.205
0.293
0.358
0.391
0.354
0.284

0.169
0.199
0.234
0.258
0.429
0.599
0.723
0.858
0.972

)(
0
1
2
3
4
5
6
7
8

09:00
10:00
11:00
12:00
13:00
14:00
15:00
16:00
17:00

)
(2

' -6 B :
7B

6B

5B

OD
4B

3B

2B

1B

37OC

37 C

37 C

37 C

45 C

RT

37 C

pH 7.0

pH 7.0

pH 10.0

pH 2.0

pH 7.0

pH 7.0

pH 7.0

NaCl 5%

Na 0.05%

NaCl 0.5%

NaC 0.5%

NaCl 0.5%

0.5% NaCl

0.5%

Cl

0.233
0.112
0.128
0.261
0.243
0.186

0.264
0.161
0.191
0.358
0.378
0.403

0.140
0.162
0.200
0.403
0.445
0.530

0.138
0.148
0.140
0.293
0.230
0.185

0.139 0.619 0.535 0.204

0.223 0.775 0.756 0.261
0.191 0.870 0.938 0.243
* : .450c -

NaCl

0.240
0.227
0.277
0.418
0.522
0.479

0.174
0.253
0.283
0.389
0.416
0.411

0.157
0.212
0.246
0.380
0.475
0.527

0
1
2
3
4
5

09:10
10:15
11:15
12:15
13:15
14:15

*
0.526
0.690
0.766

0.438
0.523
0.356

0.669
0.893
1.019

6
7
8

15:15
16:15
17:15

' -7 C :

7C

6C

5C

OD
4C

3C

2C

1C

37OC

37 C

37 C

37 C

45 C

RT

37 C

pH 7.0

pH 7.0

pH 10.0

pH 2.0

pH 7.0

pH 7.0

pH 7.0

NaCl 5%

Na 0.05%

NaCl 0.5%

NaC 0.5%

NaCl 0.5%

0.5% NaCl

0.5%

Cl

0.231
0.256
0.343
0.606
0.839
0.346

0.179
0.299
0.497
1.042
1.770
0.494

0.189
0.247
0.362
0.560
0.761
0.164

0.164
0.179
0.173
0.201
0.127
0.232

0.169
0.184
0.483
0.864
1.566
0.208

0.152
0.187
0.190
0.185
0.318
0.317

0.249
0.264
0.345
0.712
1.282
0.205

(4
0.510

(10
1.039

(10
0.524

(10
0.271

(10
0.275

0.429

(4
0.438

(4
0.911

(4
0.566

(10
0.298

(10
0.602

0.083

0.228

(5

(5
0.855

(5
0.451

(10
0.492

(10
0.588

0.377

)
(5

)
(5

0.060

NaCl

0.155

0.438

(5

(10

(10

0
1
2
3
4
5

09:10
10:10
11:10
12:10
13:10
14:10

15:10

16:10

17:10

' 8 :A1, B1, C1

)(

265000000

265

10-5

390000000

39

10-6

132000000

132

10-5

610000000

61

10-6

352000000

352

10-5

1060000000

106

10-6

90000000

10-6

1300000000

13

10-7

10-6

10-7

348000000

348

10-5

10-6

10-5

3600000000

360

10-6

10-5

4560000000

456

10-6

1930000000

193

10-6

13900000000

139

10-7

10-6

10-7

23000000

23

10-5

h0

A1

h2

h4

h6

h8

h0

B1

h2

h4

h6

h8

h0

C1

10000000

10-6

91000000

91

10-5

130000000

13

10-6

1710000000

171

10-6

10700000000

107

10-7

530000000

53

10-6

200000000

10-7

10-6

10-7

h2

h4

h6

h8

' -10 A
:
log CFU/ml
8.515211304
8.586177676
8.56937391
8.627967115
8.848804701
8.993774231
8.916333795

"
3.275E+08
3.856E+08
3.710E+08
4.246E+08
7.060E+08
9.858E+08
8.248E+08

)(
0
60
120
180
240
300
420

' -12 B
:
log CFU/ml
8.541579244
8.672015452
9.556302501
9.74515099
9.794835417
9.898450919
10.02387626

"

3.48E+08
4.70E+08

3.60E+09
5.561E+09
6.235E+09
7.915E+09
1.057E+10

)(
0
60
120
180
300
360
420

' -14 C
:
log CFU/ml
7.21748394
4
7.24288852
4
8.04336227
8
9.53733375
4
9.79274178
6
9.99660762
2
8.90255666
2

)(

"
1.650E+07

1.749E+07

60

1.105E+08

120

3.446E+09

180

6.205E+09

240

9.922E+09

300

7.990E+08

420

' -15 A ':

OD 2A
0.699
0.894
1.085
1.404
2.088
2.156
2.284
2.228
5.44

OD 1A
0.663
0.825
1.017
1.4
1.616
1.692
1.816
1.924
0.994

)(
0
60
100
140
180
220
260
300
1440

' -16 B ':

OD 2B

OD 1B

0.4
0.556
0.623
0.705
0.804
0.917
1.218
1.174
0.902

0.419
0.602
0.673
0.76
0.861
0.971
1.274
1.17
2.295

)(
0
60
100
140
180
220
260
300
1440

' -17 :A

PFU/ml
)(
3.54E+08

PFU/ml
----

PFU/ml
8.00E+08

3.58E+10

5.80E+1
0
6.90E+1
0
----

4.50E+10

4.72E+09

9.00E+0
9
0.00E+0
0

5.70E+10
1.10E+09

0.00E+00

4.50E+10
3.20E+09

0.00E+00

PFU/ml
2.30E+0
8
4.50E+0
9
0.00E+0
0
8.00E+0
7
4.30E+0
8
0.00E+0
0

PFU/ml
3.20E+07

)(
0

----

----

24

2.60E+07

----

----

24

' -18 :B
PFU/ml
)(

PFU/ml
----

0.000E+00
4.483E+09
0.000E+00

1.20E+1
0
0.00E+0
0

PFU/ml
0.00E+0
0
2.00E+0
8
0.00E+0
0

PFU/ml
0.00E+00

PFU/ml
0.00E+00

)(
0

1.25E+09

----

0.00E+00

----

24

1
0

1.00E+06

1.00E+07

----

1.44E+09

24

----

0.00E+00

1.00E+0
8
7.20E+0
9
4.88E+1
0

---3.700E+07
4.00E+0
9
4.80E+1
1

4.213E+09
2.64E+11