پایان نامه شیوع ژن های شیگا توکسین و اینتیمین در سویه های اشریشیا کلی O157H7 جدا شده از عفونت ادراری در شهرستان الشتر

فهرست:

مقدمه  ..................................................................................................................................................................   1

اهداف اصلی طرح  ..............................................................................................................................................   3

اهداف اختصاصی طرح  .......................................................................................................................................   3

اهداف کاربردی طرح  ..........................................................................................................................................   3

فرضیات یا سئوالات پژوهشی  ............................................................................................................................   4

فصل اول بررسی متون ( کلیات)  ........................................................................................................................   5

1-1 تاریخچه   History )  ).............................................................................................................................   6

2-1 طبقه بندی  classification))  .................................................................................................................   6

3- 1زیستگاه (Habitat)  ..................................................................................................................................   7

4- 1 شکل باکتری ..............................................................................................................................................   7

5-1 کشت صفات بیوشیمیابی..............................................................................................................................   7

6-1 مخازن و راه های انتقال باکتری  .................................................................................................................   8

7-1 ساختمان آنتی ژنی  .....................................................................................................................................   9

8-1 عوامل ویرولانس  ......................................................................................................................................... 11 

1_9انواع اشریشیاکلی .......................................................................................................................................... 12

10_1 باکتری اشرشیاکلی O157:H7...................................................................................... 14

ژن ها و عوامل حدت .......................................................................................................................................... 18

1_11زیستگاه طبیعی اشرشیاکلی O157:H7...........................................................................  19

1_12تشخیص .....................................................................................................................................................19

13_1 اپیدمیولوژی .............................................................................................................................................. 20

14-1 انتقال ........................................................................................................................................................ 21

15-1 علائم و نشانه ها ...................................................................................................................................... 22

16-1 تضاهرات بالینی ....................................................................................................................................... 22

17-1 درمان ...................................................................................................................................................... 24

18-1 پیشگیری ................................................................................................................................................   25

آشپزی و رژیم غذایی .......................................................................................................................................   26

فصل دوم مروری بر مطالعات گذشته  ..........................................................................................................     27

فصل سوم مواد و روش‌ها  ............................................................................................................................     32

1-3مواد و روش کار  ....................................................................................................................................     33

1-1-3 مواد  ..................................................................................................................................................     33

1-1-1-3محیط های کشت  ..........................................................................................................................     33

2-1-1-3 -سایر ترکیبات  .............................................................................................................................    33

2-1-3 –وسایل  ..............................................................................................................................................   34

2-3- جامعه آماری  .........................................................................................................................................   34

3-3- حجم نمونه  ............................................................................................................................................   34

4-3- نحوه جمع‌آوری نمونه  ...........................................................................................................................   34

5-3- کشت نمونه‌ها  ......................................................................................................................................    35

6-3- تشخیص آزمایشگاهی  ............................................................................................................................  35

1-6-3- رنگ آمیزی گرم  ................................................................................................................................  35

2-6-3- تست اکسیداز  ....................................................................................................................................  35

3-6-3- تست کاتالاز  .......................................................................................................................................35

4-6-3- تست های بیوشیمیایی  ....................................................................................................................... 35

1-4-6-3-آزمایش استفاده از سیترات  ........................................................................................................36

2 -4-6-3- آزمایش تحرک و تولید اندول  ......................................................................................................36

3-4-6-3- آزمایش متیل رد (MR )  .............................................................................................................. 37

4-4-6-3- آزمایش Voges Proskauer ( VP )  ......................................................................................37

5-4-6-3-آزمایش TSI (  Triple Suger Iron Agar )  ......................................................................... 37

8-3-  نگهداری سویه ها  ................................................................................................................................. 37

9-3-آماده سازی باکتری جهت استخراج DNA  .......................................................................................... 38

1-9-3-  تهیه محیط کشت مایع لوریا ( LB, Luria broth )................................................................38

2-9-3- کشت باکتریها بر روی محیط LB    ......................................................................................... 38

10-3- استخراج DNA به روش کیتBIONEER  ..............................................................................38

2-12-3- مراحل استخراج DNA به روش کیت BIONEER ...............................................................39

11-3- واکنش زنجیره پلیمرازی ( ( PCR  ....................................................................................................40

مواد لازم برای PCR

1-13-3 -  PCR با استفاده از این پرایمرها انجام شد  ............................................................................41

2-13-3- آماده سازی پرایمرها  ......................................................................................................................42

3-13-3- اجزای واکنش PCR  ....................................................................................................................42

4-13-3- نحوه انجام  واکنش PCR   ...........................................................................................................43

5-13-3- آماده سازی مخلوط اصلی ( Master mix )  برای واکنش  PCR  .........................................43

12-3- الکتروفورز   ......................................................................................................................................44

1-12-3- مواد مورد نیاز برای ساخت ژل آگاروز  ...............................................................................................45

2-12-3-تهیه بافر تانک الکتروفورز  ................................................................................................................45

3-12-3- روش تهیه ژل آگاروز   ....................................................................................................................45

4-12-3 طریقه رنگ آمیزی ژل  .................................................................................................................. 46

13-3- عکسبرداری از ژل  ...........................................................................................................................47

فصل چهارم نتایج  ............................................................................................................................................ 48

1-4- نتایج کشت و تعیین هویت ایزوله‌های کلینیکی........................................................................................49

2-4-نتایج بررسی های ملکولی PCR حاصل از ژن های 1stx ،2stx ، eaeA..............................................51

فصل پنجم بحث و پیشنهادات  .........................................................................................................................54

منابع  ................................................................................................................................................................. 60

منبع:

1.بنیادی م،مشتاق ح،اسفندآبادی ش،صالحی ت. 1386. مطالعه میزان آلودگی شیرهای استان چهارمحال و بختیاری به باکتری اشریشیا کلی7H157O، مجله دامپزشکی ایران،

2. دستورالعمل تشخیص آزمایشگاهی اشریشیاکلی مولد اسهال. آزمایشگاه مرجع کشوری اشریشیاکلی (آمکا)؛

3. شریفی ث. بررسی پاره ای از خصوصیات فیزیولوژیک اشریشیاکلی های جداشده از نمونه های ادرار و مدفوع جهت شناسایی سویه انتروهموراژیک O157:H7، همراه با تعیین الگوی مقاومت میکروبی باکتریهای جداشده، پایان نامه، دانشکده پزشکی کرمان، بهمن ماه ٨٠، با شماره ثبت ٢٢٧٧.

4. کارگر م و همایون م. 1388؛ بررسی شیوع منابع آلودگی سویه های انتروهموراژیک اشریشیاکلی و میزان مقاومت آن ها در مقابل آنتی بیوتیک ها در بین کودکان زیر 5 سال شهرستان مرودشت. مجله علوم پزشکی دانشگاه آزاد اسلامی، (4)19: 273-268.

5. کارگر م، حیدری س،عباسیان ف،شکرفروش ش. 1385. میزان شیوع و مقاومت آنتی بیوتیکی اشریشیا کلی 7H157O در شیر خام گاوهای شهرستان جهرم.فصلنامه بیماری های عفونی و گرمسیری وابسته به انجمن متخصصین بیماری های عفونی و گرمسیری،

6. نایب آقایی م و منصوری ش. 1385 شناسایی سویه های اشریشیا کلی O157:H مولد شیگا توکسین(STEC)  جدا شده از نمونه های مدفوع وادرار بیماران به روش  Multiplex PCR. یافته،؛ (1)8: 27-21.

7. هاشمی م، محسن زاده م، بیات م، شریعتی فر م، بنی حسن م. بررسی شیوع مقاومت آنتی بیوتیکی E. coli O157:H7 جدا شده از گوشت چرخ کرده در مشهد. مجموعه مقالات دهمین کنگره سراسری میکروب شناسی ایران، ایلام، 1388: 221.

8. بنیادی م، صالحی ت، محزونیه م، طاهری ف.1390. مطاله زن های حدت سویه های وروتوکسین زای باکتری اشریشیا کلی جدا شده از شیرهای خام و پنیر غیر پاستوریزه. مجله تحقیقاتی دام پزشکی، دوره 66 شماره 3، 228-223.

9. عرفانی ن.1389 .مقالات مروری عفونت راجعه در زنان. نشریه نوین پزشکی، شماره 490.

10. خوروش ف، زارع فر س، مصطفوی زاده ک، مباشری زاده س، ایزدی م، جنیدی ن.1386. مقایسه شیوع ارگانیسم های ایجاد کننده عفونت ادراری جانبازان مبتلا به آسیب نخاعی وبیماران دارای سوند ادراری بستری در بیمارستان الزهرای اصفهان . طب نظامی –شماره 9 (4).

11_Zilevica A 2005;  Hospital acquired and community-acquired uropathologens; modeling of infection. Bioautomation3: 63-67

12_Tanagho EA and Mc Aninch JW. Smith 2000.`s general urology. 15th Philadelphia. Mc Grow Hill.

13_Tanagho EA and Mc Aninch JW. Smith `s , 1998. general urology. 15th Philadelphia. Mc Grow Hill, 2000.Finkestein R and Kassis E. Community acquired urinary tract infection in adults: A hospital viewpoint. J Hosp Infect 38(3): 193-202.

14_ Chua MD, Annabelle T and Dytan MD.2000 Surveillance of pathogens and resistance patterns in urinary tract infections. Phil J Microbial Infect Dis; 28(1): 11

15_Lucas MJ and Cunningham FG.1993. Urinary infection in pregnancy. Clin Obstet Gynecol; 36(4): 855-868.

16_ Walker TS. 1998:. Microbiology. Philadelphia, W.B.Sanders Company, 20-30.

17_ Whittam TS, Wolfe ML, Selander RK, Dyer DW and Loos BG.1993 Clonal relationship among E.coli strains that cause hemorrhagic colitis and infantile diarrhea. Infect Immun; 61: 1619-162

18_ Adwank Abu-Hassan N, Essawi T and Bdir M.2002. Isolation and characterization of shiga toxigenic Escherichia coli strains from northern Palestine. J Med Microbiol; 51: 332-5.

19_Chart H, Perry NT, Cheasty T and Wright PA.2002. The kinetics of antibody production to antigens of Escherichia coli O157 with hemolytic uremic syndrome. J Med Microbiology; 51: 522-525.

20_Tanagho EA, McAninch JW. Smith's .2004. General Urology. 6th Edi.McCraw-Hill. 203-09.

21_Karch H, Tarr P, Bielaszewska M (2005). "Enterohaemorrhagic Escherichia coli in human medicine.". Int J Med Microbiol 295 (6–7): 405–18.

22_O antigen at Dorlands Medical Dictionary

23_Gunser F, Bohm H, Russmann H.1992. Moleculur detection of sorbitol-fermenting E.coli O157:H7 in patients with HUS, J Clin Microbial.30: 1807-100.

24_Rangel JM, Sparling PH, Crowe C, Griffin PM, Swerdlow DL. Epidemiology of Escherichia coli O157:H7 outbreaks, United States, 1982–2002. Emerg Infect Dis [serial on the Internet]. 2004 Apr [date cited]

25_Walker TS, 1998. Microbiology. Philadelphia, W.B.Sanders Company: 20-30

26_Mora A, Blanco M, Blanco JE, Dahbi G, López C, Justel P, et al. Serotypes, 2007.virulence genes and intimin types of Shiga toxin(verocytotoxin)- producing Escherichia coli isolates from minced beef in. Lugo(Spain) from 1995 through 2003. BMC Microbiology; 1(7):1-9.

27_Leotta GA, Miliwebsky ES, Chinen I, Espinosa EM, Azzopardi K, Tennant SM, et al.2008. Characterisation of Shiga toxin-producing Escherichia coli O157 strains isolated from humans in Argentina, Australia and New Zealand. BMC Microbiology;8(46):1-8.

28_Law D.2000. Virulence factors of Escherichia coli O157 and other Shiga toxin-producing E. coli. J Appl Microiol; 88(5): 729-45.

29_Karmali MA, Stede BT, Petic M, Lime.1983. Sporadic cases ofv haemoly

Tic-Uraemic sgndrome associated with faecal cytotoxin and cytotoxin-aproducing Ecoli in stools, Lancet.1:619-20.

30_Yuksel S , Ozturk B, Kavaz A, et al.2006. Antibiotic resistance of urinary tract pathogens and evaluation of empirical treatment in Turkish children with urinary tract infection. Int J Antimicrob Agent; 28(5): 413-6.

31_Razavilar, V. (1998). Microbial Pathogens in Foods, Epidemiological aspects of Food Intoxications. Tehran University press. Tehran, Iran.

32_Whittam TS, Wolfe ML, Selander RK, Dyer DW, Loos BG. Clonal 1993.relationship among E.coli strains that cause hemorrhagic colitis and infantile diarrhea. Infect Immun; 61: 1619-1629

33_Chart H, Perry NT, Cheasty T, Wright PA .2002. The kinetics of antibody.production to antigens of Escherichia coli O157 with hemolytic uremic syndrome. J Med Microbiology; 51: 522-525

34_U.S. Food and Drug Administration. Food Code: 1993 recommendations of the United States Public Health Service, Food and Drug Administration. Pub. no. PB94-11394. Washington: National Technical Information Service.

35_Blue Ribbon Task Force. Solving the E. coli O157:H7 problem. Chicago: National Livestock and Meat Board; 1994

36_Centers for Disease Control and Prevention.1997. Escherichia coli O157:H7 infections associated with eating a nationally distributed commercial brand of frozen ground beef patties and burgers—Colorado, 1997. MMWR Morb Mortal Wkly Rep.;46:777–8.

37_Bell BP, Goldoft M, Griffin PM, Davis MA, Gordon DC, Tarr PI, A multistate outbreak of Escherichia coli O157:H7-associated bloody diarrhea and hemolytic uremic syndrome from hamburgers. The Washington experience. JAMA

38_Whittam TS, Wolf ML, Wachsmuth Ik, Orskov I, Wilson RA.1993. Clonal relationships among Ecoli strains that cause hemorrhagic colitis and infantile diarrhea. Infect Immun,61:1619-29.

39_Cimolani N, Bassalygas, Mah DG, Morrison BJ, Corter JE. 1994.Acontinuling assessment of risk factor for the development of Ecoli O157:H7. Associated hemolytic uremic syndrome.Clin Nephrd,42:85-89.

40_Gliffin PM. 1995. E.coli O157:H7 and other enterohemorrhagic E.coli, In: Braster MI, Smith PD, Ruvlin JI, Greenberg Hb, Guarrant RL, eds. Infections of gasteroin testinal Tract, New York Raven Pr

41_ Riley L, Remis R, Helgerson S, McGee H, Wells J, Davis B, Hebert R, Olcott E, Johnson L, Hargrett N, Blake P, Cohen M (1983). "Hemorrhagic colitis as38sociated with a rare Escherichia coli serotype". N Engl J Med 308 (12): 681–698

42_Wells JG, Davis BR, Wachsmuth IK et al. (September 1983). "Laboratory investigation of hemorrhagic colitis outbreaks associated with a rare Escherichia coli serotype". Journal of clinical microbiology 18 (3): 512–20.

43_Doyle MP, Schoeni JL. 1984. "Survival and growth characteristics of Escherichia coli associated with hemorrhagic colitis". Applied and environmental microbiology 48 (4): 855–6.

44_whittam TS, Wachsmuth IK, Wilson RA. 1988 . "Genetic evidence of clonal descent of Escherichia coli O157:H7 associated with hemorrhagic colitis and hemolytic uremic syndrome". The Journal of infectious diseases 157 (6): 1124–33.

45_Szabo, R. A.; Todd, E. C. D.; Jean, A. 1986. "Method to isolate Escherichia coli 0157:H7 from food". Journal of Food Protection 49 (10): 768–772.

46_Reisbig R, Olsnes S, Eiklid K. 1981. "The cytotoxic activity of Shigella toxin. Evidence for catalytic inactivation of the 60 S ribosomal subunit". The Journal of Biological Chemistry 256 (16): 8739–44.

47_Calderwood SB, Auclair F, Donohue-Rolfe A, Keusch GT, Mekalanos JJ .1987. "Nucleotide sequence of the Shiga-like toxin genes of Escherichia coli". Proceedings of the National Academy of Sciences of the United States of America 84 (13): 4364–8.

48_O’Brien AD, Newland JW, Miller SF, Holmes RK, Smith HW, Formal SB. 1984. "Shiga-like toxin-converting phages from Escherichia coli strains that cause hemorrhagic colitis or infantile diarrhea". Science 226 (4675): 694–6. doi:10.1126/science.6387911.

49_Strockbine NA, Marques LR, Newland JW, Smith HW, Holmes RK, O'Brien AD .1986. "Two toxin-converting phages from Escherichia coli O157:H7 strain 933 encode antigenically distinct toxins with similar biologic activities". Infection and immunity 53 (1): 135–40.

50_Brunder W, Schmidt H, Karch H .1996. "KatP, a novel catalase-peroxidase encoded by the large plasmid of enterohaemorrhagic Escherichia coli O157:H7". Microbiology (Reading, England) 142 (11): 3305–15.

51_ Li F, Zhao C, Zhang W, Cui S, Meng J, Wu J, et al. 2005. Use of Ramification Amplification Assay for Detection of Escherichia coli O157:H7 and Other Escherichia coli Shiga Toxin- Producing Strains. J Clin Microbiol;; 43; 6086 -6090.

52_Osek J.2003. Development of a Multiplex PCR Approach for the Identification of Shiga-Toxin Producing Escherichia coli Strains and their Major Virulence Factor Genes. Journal of Applied Microbiology; 95;1217-1225

53_Bidet P, Mariani-Kurkdjian P, Grimont F, Brahimi N, Courroux C, Grimont P, et al.2005. Characterization of Escherichia coli O157:H7 Isolates Causing Haemolytic Uraemic Syndrome in France. Journal Medical Microbiology; 54; 71-75.

54_Escherichia coli". http://science.jrank.org/pages/2571/Escherichia-coli.html. Retrieved June 24, 2010.

55_Moake JL. 1994 .Haemolytic-uraemic syndrome Lancet;343:393-94.

56_Westwood ME, Whiting PF, Cooper J, et al.2005 . Further Investigation of Confirmed Urinary Tract Infection (UT1) in Children Under Five Years; a Systematic Review. BMC Pediatr; 5: 2.

57_Mead PS. 1998. Griffin Pm, Escherichia coli O157:H7, Lancet;352:1207-120.

58_Tiden J, Young W, MC Namara AM.1996. A new route of Transmission for E.coli O157:H7. Am J Publ H 1th; 86:1142-45.

59_Lesaux N, Spika JS, Friesen B. 1993. Groand  beef consumption in noncommercial setting is a risk factor for sporadic E.coli O157:H7.Infection in Canada .J Infect Dis;167:500-02.

60_ J.D. Greig, E.C.D. Todd, C. Bartleson, and B. Michaels. 2010. "Infective Doses and Pathen Carriage", pp. 19-20, USDA 2010 Food Safety Education Conference.

61_Brazil, S. M.; Steelman, C. D.; Szalanski, A. L. 2007. "Detection of pathogen DNA from filth flies (Diptera: Muscidae) using filter paper spot cards". Journal of Agricultural and Urban Entomology 24 (1): 13–18.

62-Szalanski, A. L.; Owens, C. B.; McKay, T.; Steelman, C. D. 2004. "Detection of Campylobacter sp., and E. coli O157:H7 in filth flies (Diptera: Muscidae) by polymerase chain reaction".Medical and Veterinary Entomology 18 (3): 241–246.

63_Slutsker L, Riec AA, Greene KD, Hutwagner L, Griffin PM.1997. Ecoli O157:H7 diarrhea in the USA. Clinical and epidemiological feature:Ann Intern Med;126:1207-120.

64_Ban on E. Coli in Ground Beef Is to Extend to 6 More Strains". New York Times. September 12, 2011. Retrieved 2011-10-08. "After the U.S.D.A. banned the O157 form of E. coli from ground beef in 1994, the meat industry sued to block the move, but the agency prevailed in court.

65_Todd W.T.A, Dundas S.2001. The mangment of VTEC O157 Infection. Inter J Food Nicro; 66: 103_ 110.

66_Bell BP, Griffin PM, Lozano P.1997.predictors of HUS in children during a large outbreak of E.coli O157:H7 in factions pediatrics; 100: 12-16.

67_Murakami J, Kishi K, Hiramutsu K. Macrolides and clindamycin.2000 .suppress relese of shiga like toxins from E.coli O157:H7, Inter J Antimicro Agents; 15 : 103-109.

68_Nathan M, Thielman, Richard L, Guerrant.2004. Acute Infections Diarrhea, N Engl J Med; 350:38-47

69_James Chin.2000. Diarrhea caused by Escherichia coli, Control of Communicalble Diseases, 17 th ed.American public Health Association , PP.155-158.

70_Walterspiel JN, Ashkenazi S, Morrow AL, Cleary TG. 1992. "Effect of subinhibitory concentrations of antibiotics on extracellular Shiga-like toxin I". Infection 20 (1): 25–9.

71_Sutsker L, Allen A, Katherine D, Joy G, Lori H and Patrica M.1997. Escherichia coli O157:H7 diarrhea in the united States: clinical and Epidemiologic Features Annals of Internal Medicine, 126: 505_513

72_Mead PS, Slutsker L, Dietz V, MCocaing LF, Bresee JS, Shapiro C. Food related illness and death in the United States Emerg Infect. Dis, 5:607-625.

73_Nataro JP and Kaper JB.1998. Diarrheagenic Escherichia coli. Clin Microbiol Rev; 11(1): 142-201.

74_Riley LW, Remis RS, Helgerson SD, McGee HB, Wells JG, Davis BR, Hebert RJ, Olcott ES, Johnson LM, Hargrett NT, Blake PA and Cohen ML. 1983. Hemorrhagic colitis associated with a rare Escherichia coli serotype. N Engl J Med; 308(12): 681-685

75_Hogan MC, Gloor GM, Uhl JR, Cockerill FR and Milliner DS. 2001. Two cases of non-O157:H7 Escherichia coli hemolytic uremic syndrome caused by urinary tract infection. Am J Kidney Dis38:E22.

76_Johnson JR, Jerome C, Boster DR, Stapleton AE and Tarr PI.2002. Analysis of Urinary Escherichia coli Isolates for Ability To Produce Shiga Toxin. J Clin Microbial; 40(6): 2247-2248.

77_Forfar J and Arneil G.1998. Diseases Due to Infection. Text book fo pediatrics. 3 rd ed. London Churchill Living stone: 949.

78_Coia JE,1998. Microbiological and epidemiological aspects of E.coli O157:H7 in Infection , FEMS Immu and MED Micro (20)1-9.

79_Zamir G, Sakran W, Horowitz Y, et al.2004. Urinary Tract Infection Arch Dis Child; 89: 466–468

80_Murphy B P , Murphy M , Buckley J F .2005. In –Line milk filter analysis : Escherichia coli O157:H7 surveillance of milk production holding . Int J Hyg

Environ – Healthy 4521-4528.

81_Chang J H , Chou C.2000. Growth and survival of Escherichia coli O157:H7 during the fermentation and storage of diluted cultured milk drink . Food Microbiology; 17: 579-587.

82_Widiasih D A , Ido N , Omoe K .2003. Duration and magnitude of faecal shedding of shiga toxin – producing Escherichia coli from naturally infected Cattle . Epidemiol . Infect . 132 : 67-75.

83_Paton AW, Paton JC.1998. Detection and characterization of Shiga toxigenic Escherichia coli by using multiplex PCR assay for stx1, stx2, eaeA, enterrohemorrhagic Escherichia coli hlyA, rfbO111, rfbO157. J Clin Microbiol 36(2):598-602.

84_Pradel N, Liverlli V, Champs CD, Palcoun J, Reynaud A, Scheutz F, et al.2000 . Prevalence and characterization of Shiga-toxin producing Escherichia coli isolated from cattle, food, and children during a one-year prospective study in France. J Clin Microbiol.38(3):1023-31.

85_Blanco M, Blanco JE, Mora A, Rey J, Alonso JM, Hermoso M, et al. 2003. Serotypes, virulance genes, and intimin types of shiga toxin (verotoxin)- producing Escherichia coli isolates from healthy sheep in spain. J Clin Microbiol 41(4):1351-6.

86_Mohsin M, Hussain A, Butt MA, Bashir S, Tariq A, Babar S, et al. 2007. Prevalence of Shiga toxin- producing Escherichia coli in diarrheal patients in faisalabad region of pakistan as determined by multiplex PCR. J Infect Developing Countries.1(2):164-9.

87_Estrada-Garcia T, Cerna JF, Paheco-Gil L, Velazquez RF, Ochoa TJ, Torres J, et al.2005. Drug- resistant diarrheogenic Escherichia coli , Mexico. Emerging Infectious Disease .11(8):1306-8.

88_Santaniello A, Gargiulo A, Borrelli L, Dipineto L, Cuomo A, Sensale M, et al.2007. Survey of Shiga-toxinproducing Escherichia coli in urban pigeons (Columba Livia) in the city of Napoli, Italy. Ital J Anim Sci .6(2): 313-6.

89_Gannon VPJ, King RK, Kim JY and Thomas EJG.1992. Rapid and Sensitive Method for Detection of Shiga-Like Toxin-Producing Escherichia coli in Ground Beef Using the Polymerase Chain Reaction. J Appl Environ Microbial.58(12): 3809-3812.

90_Aslani MM, Bouzari S.2003. An epidemiological study on verotoxin-producing Escherichia coli infection among population of northern region of Iran. Eur J Epidemiol.18(4).

91_Alikhani MY, Mirsalehian A, Fatollahzadeh B, Pourshafie MR and Aslani 2007.MM. Prevalence of enteropathogenic and shiga-toxin producing Esscherichia coli among children with or without diarrhea in Iran. J Health Popul Nutr. 25 (1): 88-93.

92_S. M. Hosseini, H. Ezzatpanah, M. Aminlari, M. Mazaheri Assadi,D. Ataie.2011. Investigating the Contamination of E.coil O157:H7 in Processed Meat Products Produced in Two Factories at Shiraz and Tehran. Food Technology & Nutrition / Summer .Vol. 8 / No. 3

93_Jafareyan-Sedigh MR, Rahimi E, Doosti A 2011.Isolation of Escherichia coli O157: H7 in sheep meats using cultural and PCR method.June, July; 13(2).

94_Bonyadian, M, Zahraei Salehi, T, Mahzounieh, M .R, Akhavan Taheri. F.

2011.VIRULENCE GENES OFVEROTOXIGENIC E. COLI ISOLATED FROM RAWMILK AND UNPASTURIZED CHEESE.J.Vet.Res. 66,3:223-228

95_Pollard DR, Tyler SD, Rozee KR, Johnson MW and Lior H.1990. Rapid and specific detection of verotoxin genes in Escherichia coli by the polymerase chain reaction. J Clin microbial. 540-545.

96_Baron E, Finegold SM.1994.Diagnostic Microbiology.9th  Edition, Mosby, Philadelphia.

97_Adwank Abu-Hassan N, Essawi T, Bdir M. 2002. Isolation and characterization of shiga toxigenic Escherichia coli strains from northern Palestine. J Med Microbiol 51: 332-5

98_Kim JY, Kim S, Kwon N, Bae WK, Lim JY, Koo HC, et al.2005. Isolation and identification of Escherichia colio157:h7 using different detection methods and molecular determination by multiplex PCR and RAPD. J Vet Sci.6(1):7-19.

99_S. M. Hosseini, H. Ezzatpanah, M. Aminlari, M. Mazaheri AssadiD. Ataie .2011. Investigating the Contamination of E.coil O157:H7 in ProcessedMeat Products Produced in Two Factories at Shiraz and Tehran. Food Technology & Nutrition / Summer .Vol. 8 / No. 3

100_Josefa M, Rangel T, Phyllis H, Patricia M and David L.1982-2002 .Epidemiology of Escherichia coli O157:H7 outbreaks, United States, volume 11,N.

101_ Rily IW,Remis Rs, Helgerson SD. 1983.Hemorrhagic colitts associated with a rare Escherichia coli serotgpe.N Engl J Med,308:681-685.

101_Jazayeri M. 2000. Frequency of bacterial agents in urinary tract infection and their and antibiotic susceptibility pattern in Semnan. Journal of Semnan University of Medical Sciences, Vol. 1, No.4.

102_Santaniello A, Gargiulo A, Borrelli L, Dipineto L, Cuomo A, Sensale M, et al.2007. Survey of Shiga-toxin producing Escherichia coli in urban pigeons(Columba Livia) in the city of Napoli, Italy. Ital J Anim Sci 6(2): 313-6