effect of bacteriophage on proteus

1
??? ???? ?????? ??????   ((?????????? ???? ??????
????? ????? ?????)) ??? ???? ?????? ?????? ????
???????? ????? 30
2
Evaluation of bacteriophage effect against
multidrug-resistant Proteus mirabilis isolated
from different clinical samples
????? ????? ???? ??????? ??? ?????? ??????
?????? ??? ???? ??????

• ????? ???? ??? ?????? ????

3
??????? Introduction
4
???????

• ??? ??????? Proteus mirabilis ????? ????????
  ???????? ????? ????? ???? ?????? ????? ???
  (1-3) ?????????? ?????? (0.8 -0.4) ???????????
  ???? ???? ??? ?? ??? ?????? ???????? Hauser ???
  1885? ?????? ??????? ??? ??????? ??? ?????
  ??????? ????? ???????? ????? ???????? Swarming
  ??? ????? ????? ???? ????????? ????? ??????
  Peritrichous Flagella ??? ?????? ????????? ???
  ????? ????? ??????? Urease.

5
????? ???????

• ????? ??????? P.mirabilis ?????? ?? ????? ???????
  ????? ????? ??????? ?? ???? ??? ????? ??????? ??
  ??????? ?????? ???? ?????? ???????? ?????????
  ????? ???????? ??? ????? ??????? ??? ?????? ???
  ?????? ?????? ??? ????? ????? ???????? Swarming
  ?? ???? ?? ??????? ????????? ????? ??? ?????????
  ?? ??????? ???????? ?????? ??????? P.mirabilis
  ????????? ??? ????? ?????? ?????? ???????
  Crystalline Biofilm ??? ??????? ???????? ???????
  ??????? ???????? ??? ????? ????? Protease ????
  ??? ??? ????????????? ???? ???????? ?????????
  ??????? ?? ?????? ??????? ?????? ?? ???? ???
  ????? ????????????? ????????.

6
?????????

• ????? ??????? P.mirabilis ??? ????? ????????
  Opportunistic ??? ??? ???? ?????? ?? ??????? ???
  ???? ?? ??? ??????? ??? ??? ??? ??????? ??????
  ??????? ??????? ????? ???? ?????? ????? ??????
  ?????? ????.

???????? ???????? ???????
??? ??????? P.mirabilis ??? ?????? ??????
???????? ??????? ?? ????? ?????? ????? ???????
???????????? ??? 1990 ????? ????????? ???????
?????????????? ????? ????????? ??? ????? ???????
???????? ??????? ???? ???? ?????? ???? ??????
????????? ????? ??? ???? ?????? ??????
7
?????? ???????? Bacteriophage

• ??? ?????? ???????? ???? ??? ??? 1915 ?? ???
  ?????? Frederick Twort ??? ??? 1917 ?? ??????? ??
  ??? ?????? ??????? Felix dHerelle ????? ??? ??
  ??????? ?????? ???????? ?? ???? ??? ????? ??????
  ?? ??????? ? ????? ?????? ???????? ?? DNA ?? RNA
  ???? ????? ??????? ???? ???? ??????.

8
????? ???????

• ??? ?????? ??????? P.mirabilis ????????? ???
  ??????? ???????? ????????? ??????? ??????????? ??
  ????? ?????? ??????.
• ????? ??? ???????? ???????? ??????? ??????
  ??????? ?????? ??????.
• ????? ??????? ?? ??????? ?????????????? ????????
  ?????????????? ????? ?????.
• ????? ?? ???? ????????? ??? ????? ?????? ??????
  Biofilm ??????? ??????? ????????? ????????
  ???????.
• ??? ?????? ?????? ???????? Bacteriophage ??????
  ?????? ??? ??????? P.mirabilis ??? ????????
  ???????? .

9
?????? ?????? ????? Materials and Methods
10
(No Transcript)
11
????? ???????
Biofilm
Protease
Urease
Microtiter plate
Urea agar
Skim milk agar
12
(No Transcript)
13
????? ?? ??????? ?????????????? ????????
?????????????? ????? ?????
????? ?? ??????? ??????????????
????? ?? ??????? ?????????????? ????? ?????
Iodometric method
Double disk synergy
14
??? ?????? ?????? ????????
1- ??? ???????
2- ??? ?????? ????????
3- ????? ?????? ????????
4- ????? ????? ?????? ???????? ??? ?????????
5- ????? ????? ?????? ?? ????? ??????? ?????
??????????? ??? ?????? ????????
15
??????? Results
16
??? ?????? ??????? P.mirabilis

• ???? ????????? ???????? ??? ?????? ???????
  Cultural Characterizes ????????? Microscopy
  ????????? ???????????Biochemical Tests ?? ??
  ?????? ??? 30 ???? ???? ??? ??????? P.mirabilis
  ????? ???? ??????? ???? ?? ?????? ????? ??
  ??????? Urine 70 (21) ??? ????? Sputum ???????
  Wounds 10(3) ??? ????? ????? ?? ???????? Ulcer
  4 (1) ????? ???? ?? ????? ????? Pus ??? ??????
  ????? ?????? Otitis media 3(1) ??? ????? ??? ??
  ????? ?????

17
??????? ??????? ? ???????

• ???????- ???? ??????? ??? ??? ???? ????? Blood
  agar ???? ????????? ?????MacConkey agar ???? ???
  ???? ????? ??????? ????? Brain-Heart infusion
  agar ???? ?????????? ???????? CHROMagar
  orientation ????? ????? ????? 37 º? ???? 24
  ????? ????? ???? ???? ???????? Swarming ??? ???
  ???? ???? ???? ????? ??????? ????? ????? ???????
  P.mirabilis ??? ??? ????????? ????? ???? ????
  ????Pale Yellow ?????? ??? ????? ???? ????????
  ??? ????? ???????? ???? ???? ????? ????? ????????
  Moldy Fish ????? ???????? ??????? P.mirabilis ???
  ??? ?????????? ???????? ???? ???.
• ??????? - ???? ???? ?? ??????? P.mirabilis ?????
  ????? ????? ???? ????????? ???? ????? ????? ?????
  ????? ????.

18
??????? ??????????

• ????? ??????? ??? ???? ????? ??????? P.mirabilis
  ????? ???? ?????????? Oxidase test ???? ????
  ???????? Voges-Proskauer test ???? ??????? Indole
  test ????? ????? ???? ???? ?????? Methyl Red test
  ???? ???????? Catalase test ????? ??????? ????
  ??????? ??????? Citrate Utilization test ?????
  ???? ?? ??? ???? ???????? ??? ??? Kligler Iron
  agar ????? ????? ???? ???????? ?????? ???? H2S
  ??????? ?????????? ??? ????? ??????? ???? ?????
  ?????? ???????? CO2

19
?????? ???????? ???????? ???????

• ????? ???????? ???????? Disk Diffusion Method
• ?? ????? ??? ???????? ????? 30 ???? ?? ???????
  P.mirabilis ???????? 10 ?????? ????? ?????? ????
  ??????? ?????? ????? ????? 76.6 ?????
  Cefotaxime? ?????? 66.7 ??? ?? Tobramycin
  ?Amikacin ?? ??? ???? ???? ???????? ?????
  Gentamicin 60? ??? ?????? ?????? ?????
  Ceftazidime ????? Ciprofloxacin ????? 40 ???
  ?????? ????? ??????? ????? ?????? ?? ??????
  ????????? ???????? Imipenem? Norfloxacin?
  Amoxicillin/Clavulanic acid ? Aztreonam ??????
  36.7? 33.3? 30 ? 23.3 ??? ???????.

20
????? ???? ??????? ?????? ??????

• ????? ???? ??????? ?????? ?????? MIC????????
  ??????? ???????? ???? ???????
• ?? ??????? 17 ?????? ?????? ?????? ???? ???????
  ?????? ??????. ?? ????? ????????? ?????? ?????
  ????? ?? ???? Tigecycline ?Nitrofurantoin ?????
  100? ?????? 83.4 ???????? Ampicillin?
  Cefazolin? Ceftriaxone? Cefepime? Aztreonam
  ?Imipenem? ????? ???? ???????? ??????
  Trimethoprim/Sulfamethoxazole 76.6 ??????
  Moxifloxacin 50 ?????? ??????? ?????? ??????
  ?????? 40 ????? Ampicillin/Sulbactam ?????
  Ciprofloxacin ?????? 30 ????? Gentamicin? ?? ???
  ???? ???? ???????? ????? Tobramycin 16.6 ??????
  Amikacin 3.34. ?? ??? ?? ???? ????????? ??????
  ????? ?? ???? Ertapenem ????? Meropenem.

21
???? ????? ??????? ??????? ??????? P.mirabilis
???? ??????? ?????? ?????? MIC ???????? ????
???????
22
???????? ???????? ???????? ???????
23
????? ??????? ?????????????? ???????????????
????? ?????

• ???? ????? ??????? ??????? ??? 11 ???? ???? ?????
  ???????? ?????????????? ????? 36.7 ?? ??? ?????
  ????? ??????? ??????? ??? 8 ????? ?? ???????
  P.mirabilis ?????? 26.7 ??? ?????? ??? ?????
  ??????? ?????????????? ????? ?????

24
????? ???????

• ????? ????? ??????? Urease Production
• ????? ????? ??????? ??????? ????? ???? ?????
  ??????? P.mirabilis ??? ????? ????? ??????? ?????
  100? ????? ????? ??? ???? ??????? ?? ???? ???
  ??????? ???? ??? ?????? ??? (4-6) ????? ?? ????
  ???????
• ????? ????? ????????? Protease Production
• ?? ????? ?? ????? ????????? ???? ???????? ???
  ?????? ???? ????? ????? Skim Milk Agar ?????
  ????? ??????? ??? 90 ?? ????? ???????
  P.mirabilis ??? ?????? ??? ????? ???????.

25
????? ???????

• ????? ?????? ?????? Biofilm Formation
• ?? ????? ?? ?????? ????? ??????? P.mirabilis ???
  ????? ?????? ?????? ?? ?????? ????? ???????
  ???????? Microtiter plate? ????? ??????? ???????
  ??????? ???????? ??? ???? ELISA ????? ???? 630
  ???????? ???? ???? ??????? ??????? ?????? ???????
  P.mirabilis ?????? ??? (0.014-0.163).????? ?????
  ??????? ??????? ?? ???? ????? ??????? P.mirabilis
  ??? ?????? ??? ????? ?????? ?????? ?????? ?????
  ??????? ???? ???? ??????? ??????? ??????? ???????
  (0.002).

26
??? ?????? ???????? Isolation of Bacteriophage
????? ????? ???????? ??? ??????? ???? ?? ????? ??
????? ?????? ????? ??????? ????? ??? ???? ??????
?????? ??? ????? ?? ???????? ??????? ??? ??????
??? ????? ???? ????? ?? ??????? P.mirabilis ???
???????? ???????? ???????.
27
???? ?????? ????????Characterizes of
Bacteriophage
?? ????? ???? ?????? ???????? ?? ???? ???? ???
???? ??????? Plaques ???? ?????? ?????? ????????
??? ??? ??????? ?? ???? ????? ??????? ??????? ???
??? Plaques ?????? ??? 0.5-3 ?????? ????? Plaques
????? ???????? ???????? ?????? ?? ??? ?????
??????? ????? ?? ??? Plaques ????????
28
????? ?????? ?? pH ???????? ?? ?????? ????????
???????
pH
????? ???????? ????????? ???????? ?????? ???
????? ????????? ?? ????? ??????? ???????? ???????
????? ???? ??? ????? ???? ???? ?? ????? ???????
(25? 37? 45? 55)º?? ????? ?? ???? ????? 65 º? ???
???? ?????? ???????? P.mP20 ?????? ??? ?????
??????? P.mirabilis ??? ?????? ??????
?????? ??? pH (2? 4? 7? 9? 11) ?????? ??????
?????? ???????? ??? ????? ????????? ?? ?????
?????? ?? pH? ?????? ????? ??????? ??????? ???
???? ???????? ????????? ???????? ??? ?????
????????? ????? ???? ???? ???? ?? pH (2)? ?? ???
??? ?????? P.mP21 ?? ?????? ??? ????? ?????????
?? pH (4) ?pH(11)? ????? ????? ???? ????????
?????? ??? ????? ????????? ?? pH (7) ?pH (9)
29
???? ????? ????? ??????? ?pH ??? ????????
????????? ????????
30
??????????? ? ????????Conclusions Reconditions
31
???????????

1. ????? ????? ??? ??????? P.mirabilis ????? ??????
   ????? ????? ?? ????? ???????? ??????? ???????
   ???????.
2. ???? ???? ????? ??????? P.mirabilis ????????
   ??????? ????? Tigecycline ????? Nitrofurantoin
   ?????? 100? ?? ??? ????? ?????? ????? ?????
   Ertapenem ????? Meropenem ?????? 100.
3. ????? ??????? P.mirabilis ???????? ??? ?????
   ??????? ?????????????? ???????? ??????????????
   ????? ????? ?????? 36.7 ?26.7 ???? ???????.
4. ????? ????? ??????? ??????? ?????? ???????
   P.mirabilis ??? ????? ?????? ?????? ?????? ?????
   ??????? ?????? 100? ?? ??? ???? ???? ????? ?????
   protease 90.
5. ?? ??? ????? ?????? ??????? ????? ???????? ???
   ??? ??????? P.mirabilis ??? ???????? ????????
   ???????? ???????.
6. ????? ????? ????? ???? pH ???????? ??? ????????
   ????????? ?????? ???????? ??? ??? ????????? ???
   ??? pH (7-9) ??????? ????? ?? ??? ???? ????????
   ????? ??? ??? ????????? ??? ????? ??????? (25-55)
   º?.

32
????????

• ????? ?????? ?? ???????? ?? ?????? ?????????
  ???????? ??????? ????? ?? ???????? ???????.
• ????? ??????? ??????? ?????? ???????? ????????
  ?????? ??????????.
• ????? ?????? ?? ??????? ??? ???????? ?????????
  ????????? ?? ?????? ??????? ???? ??????? ??
  ???????? ???????.
• ????? ?????? ?? ???????? ?? ??? ???????? ???????
  ?? ?????? ???????? ?????? ?????? ?????? ??
  ????????? ???????.
• ????? ?????? ?? ????? ?????? ???????? ?? ?????
  ??????? ?? ????????? ???????.
• ????? ??????? ?????? ?????? ????????.

33
????? ???? ???????