ردیابی ژنهای حدت شیگاتوکسین و اینتیمین در اشریشیاکلی جدا شده از یک کبوتر با علایم گوارشی: گزارش موردی
محورهای موضوعی :
مجله پلاسما و نشانگرهای زیستی
مجید غلامی آهنگران
1
,
مهرداد استادپور
2
,
آنیتا خانی
3
1 - بخش بهداشت و بیماری های طیور، دانشکده دامپزشکی، واحد شهرکرد، دانشگاه آزاد اسلامی، شهرکرد، ایران
2 - دانش آموخته دانشکده دامپزشکی، واحد شهرکرد، دانشگاه آزاد اسلامی، شهرکرد، ایران.
3 - دانش آموخته دانشکده دامپزشکی، واحد شهرکرد، دانشگاه آزاد اسلامی، شهرکرد، ایران
تاریخ دریافت : 1399/08/29
تاریخ پذیرش : 1400/02/22
تاریخ انتشار : 1401/09/01
کلید واژه:
اﻳﻨﺘﻴﻤﻴﻦ,
ﺷﻴﮕﺎﺗﻮﻛﺴﻴﻦ,
اﺷﺮﻳﺸﻴﺎﻛﻠﻰ,
ﻫﻤﻮﻟﻴﺰﻳﻦ,
کبوتر,
چکیده مقاله :
زمینه و هدف: اشریشیاکلی به عنوان فلور در دستگاه گوارش تمام خونگرمها و از جمله کبوتر یافت میشود. این باکتری دارای ژنهای حدت زیادی است که از جمله میتوان به شیگاتوکسین و اینتیمین اشاره کرد. اینتیمین مسئول اتصال اشریشیاکلی به سلولهای مخاطی روده بوده و شیگاتوکسین باعث مرگ سلول میشود. هدف از مطالعه اخیر بررسی حضور این ژن های حدت در اشریشیاکلی جداشده از یک مورد عفونت گوارشی در کبوتر به منظور امکان بیماریزایی اشریشیاکلی با منشا کبوتر در انسان می باشد.
مواد و روش ها: سوآب کلواکی از یک کبوتر با علایم گوارشی ارجاع شده به کلینیک دامپزشکی تهیه شد. ﭘﺲ از ﻛﺸﺖ و ﺧﺎﻟﺺﺳﺎزى، ﺑﺎ ﺗﺴﺖﻫﺎى ﺑﻴﻮﺷﻴﻤﻴﺎﻳﻰ ﺑﻪ ﺗﺄﻳﻴﺪ ﻣﻮارد آﻟﻮدﮔﻰ اﺷﺮﻳﺸﻴﺎﻛﻠﻰ ﭘﺮداﺧﺘﻪ ﺷﺪ. ﺳﭙﺲ ﺑﺎ اﺳﺘﺨﺮاج DNA ﺑﻪروش ﺟﻮﺷﺎﻧﺪن، ﺑﻪ ردﻳﺎﺑﻰ ژنﻫﺎى ﻫﻤﻮﻟﻴﺰﻳﻦ، اﻳﻨﺘﻴﻤﻴﻦ و زﻳﺮ واحدﻫﺎى ١ و ٢ ﺷﻴﮕﺎﺗﻮﻛﺴﻴﻦ (stx1 و stx2) ﭘﺮداﺧﺘﻪ ﺷﺪ. ژن های مورد نظر با پرایمر های اختصاصی و براساس تکثیر ژن های هدف با طول قطعه مشخص (به ترتیب 165، 890، 614 و 779 جفت بازی) مورد شناسایی قرار گرفتند.
نتایج: ﻧﺘﺎﻳﺞ ﻧﺸﺎن داد سویه اشریشیاکلی جدا شده از کبوتر با علایم اسهال واجد ژنهای حدت شیگاتوکسین 1 و اینتیمین بوده و فاقد ژن کد کننده همولیزین و شیگاتوکسین 2 میباشد.
نتیجه گیری: نتایج نشان داد ژنهای حدت شیگاتوکسین 1 و اینتیمین در کبوتر واجد علایم گوارشی قابل ردیابی است. لذا با توجه به این نتایج، بر خلاف تصورات قبلی، ممکن است پرندگان منشا اشریشیاکلی شیگاتوکسوژنیک باشند و ممکن است علایم اسهال در کبوتر با حضور اشریشیاکلی واجد ژن شیگاتوکسین و اینتیمین در ارتباط داشته باشد که لازم است برای اثبات آن بررسیهای بیشتری انجام شود.
چکیده انگلیسی:
Background & Aim : Escherichia coli (E. coli) is found as a flora in the digestive tract of all warm-blooded animals, including pigeons. This bacterium has many virulent genes including shigatoxin and intimin. Intimin is responsible for binding E. coli to intestinal mucosal cells, and shigatoxin causes cell death. The aim of recent study was to investigate the presence of these virulence genes in E. coli isolated from a case of gastrointestinal infection in pigeons in order to determine the possibility of pathogenic E. coli with origin of pigeons in humans.
Materials and Methods: In this regard, to identify virulent genes in E. coli isolated from diarrheic pigeon, a cloaca swab was prepared from a pigeon with diarrhea symptoms. After microbial culture and purification, the infection to E. coli was confirmed by biochemical examinations. Then, DNA was extracted by boiling method and hemolysin, intimin and sub unites of shigatoxin (stx1 & stx2) genes were detected. The target genes were identified by specific primers based on amplification of target genes with specific fragment lengths (165, 890, 614 and 779 bp, respectively).
Results: The E. coli strain isolated from pigeon with symptom of diarrhea had the virulence genes of stx1 and intimin and lacks the genes encoding hemolysin and stx2.
Conclusion: The results showed that the virulence genes of shigatoxin 1 and intimin were detectable in pigeons with gastrointestinal symptoms. According to these results, contrary to previous perceptions, birds may be the source of shigatoxogenic E. coli, and the symptoms of diarrhea in pigeons may be related to the presence of Shigatoxin and intimine in E. coli, which needs to be further investigated.
منابع و مأخذ:
1. Adeli, Z., Firoozeh, F., Zibaei, M., Shakib, P. )2013(. Prevalence of Shiga toxin and Intimine genes in Shiga toxin-producing Escherichia coli isolated from urine samples in Lorestan, Iran. FEYZ, 17(2); 188-194.
2. Awad-Alla, M.E., Abdien, H.M., Dessouki, A.A. )2010(. Prevalence of bacteria and parasites in White Ibis in Egypt. Vet. Ital., 46(3); 277-86.
3. Byrne, L., Vanstone, G.L., Perry, N.T., Launders, N., Adak, G.K., Godbole, G., et al. )2014(. Epidemiology and microbiology of Shiga toxin-producing Escherichia coli other than serogroup O157 in England, 2009–2013. J. Med. Mic., 63(9); 1181-1188.
4. Croxen, M.A., Law, R.J., Scholz, R., Keeney, K.M., Wlodarska, M., Finlay, B.B. )2013(. Recent advances in understanding enteric pathogenic Escherichia coli. Clin. Mic. Rev., 26(4); 822-880.
5. Estrada-García, T., Cerna, J.F., Paheco-Gil, L., Velázquez, R.F., Ochoa, T.J., Torres, J., et al. )2005(. Drug-resistant diarrheogenic Escherichia coli, Mexico. Emerg. Infect. Dis., 11(8); 1306.
6. Fagan, P.K., Hornitzky, M.A., Bettelheim, K.A., Djordjevic, S.P. )1999(. Detection of Shiga-like toxin (stx1 andstx2), intimin (eaeA), and enterohemorrhagic Escherichia coli (EHEC) hemolysin (EHEC hlyA) genes in animal feces by multiplex PCR. Appl. Environ. Microbiol., 65(2); 868-872.
7. Friesema, I., Van Der Zwaluw, K., Schuurman, T., Kooistra-Smid, M., Franz, E., Van Duynhoven, Y., et al. )2014(. Emergence of Escherichia coli encoding Shiga toxin 2f in human Shiga toxin-producing coli (STEC) infections in the Netherlands, January 2008 to December 2011. Eurosurveillance., 19(17(; 20787.
8. Gholami-Ahangaran, M., Zia-Jahromi, N. )2014(. Identification of shiga toxin and intimin genes in Escherichia coli detected from canary (Serinus canaria domestica). Toxicol.Indust. Health., 30(8); 724-727.
9. Gioia-Di Chiacchio, R.M., Cunha, M.P.V., Sturn, R.M., Moreno, L.Z., Moreno, A.M., Pereira, C.B.P., et al. )2016(. Shiga toxin-producing Escherichia coli (STEC): Zoonotic risks associated with psittacine pet birds in home environments. Vet. Microbiol., 184; 27-30.
10. Guenther, S., Ewers, C., Wieler, L.H. )2011(. Extended-spectrum beta-lactamases producing coli in wildlife, yet another form of environmental pollution. Front. Microbiol., 2; 246.
11. Haag-Wackernagel, D., Moch, H. )2004(. Health hazards posed by feral pigeons. J Infect., 48(4); 307-313.
12. Hosseini, S., Ranjbar, R., Zahrayi Salehi, T. )2016(. The Identification of pathogenic Escherichia coli strains to carry intimin gene isolated from various water sources. Iran. J. Med. Microbiol., 10(4); 17-24.
13. Kimpe, A., Decostere, A., Martel, A., Haesebrouck, F., Devriese, L.A. )2002(. Prevalence of antimicrobial resistance among pigeon isolates of Streptococcus gallolyticus, Escherichia coli and Salmonella enterica serotype Typhimurium. Avian Pathol., 31(4); 393-397.
14. Mora, A., Blanco, M., Blanco, J.E., Dahbi, G., López, C., Justel, P., et al. )2007(. Serotypes, virulence genes and intimin types of Shiga toxin (verocytotoxin)-producing Escherichia coli isolates from minced beef in Lugo (Spain) from 1995 through 2003. BMC Microbiol., 7(1); 13.
15. Morabito, S., Dell’Omo, G., Agrimi, U., Schmidt, H., Karch, H., Cheasty, T., et al. )2001(. Detection and characterization of Shiga toxin-producing Escherichia coli in feral pigeons. Vet. Microbiol., 82(3); 275-283.
16. Nolan, L.K., Barnes, H.J., Vaillancourt, J.P., Abdul-Aziz, T., Logue, C.M. )2020(. Colibacillosis. In: Disease of Poultry. Swayne, D.E., Boulianne, M., Logue, C.M., McDougald, L.R., Nair, V. and Suarez, D.L. editors. 14th ed., Massachusetts: Wiley-Blackwell, pp: 770-790.
17. Paton, J.C., Paton, A.W. )1998(. Pathogenesis and diagnosis of Shiga toxin-producing Escherichia coli Clin. Microbiol. Rev., 11(3); 450-479.
18. Persad, A.K., Lejeune, J.T. )2015(. Animal reservoirs of Shiga toxin-producing Escherichia coli. In Enterohemorrhagic Escherichia coli and Other Shiga Toxin-Producing E. coli. Am. Soc. Microbiol., 231-244.
19. Sandvig, K. )2001(. Shiga toxins. Toxicon, 39(11); 1629-1635.
20. Schmidt, H., Scheef, J., Morabito, S., Caprioli, A., Wieler, L.H., Karch, H. )2000(. A new Shiga toxin 2 variant (Stx2f) from Escherichia coli isolated from pigeons. Appl. Environ. Microbiol., 66(3); 1205-1208.
21. Ziauddini, A.H., Gholami-Ahangaran, M., Ahmadi-Dastgerdi, A. )2020(. Detection of virulence genes of intimin, hemolysin and shigatoxin in Escherichia coli isolated from Pscittacin. New Cell. Molecul. Biotechnol. J., 10(38); 61-68.
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1. Adeli, Z., Firoozeh, F., Zibaei, M., Shakib, P. )2013(. Prevalence of Shiga toxin and Intimine genes in Shiga toxin-producing Escherichia coli isolated from urine samples in Lorestan, Iran. FEYZ, 17(2); 188-194.
2. Awad-Alla, M.E., Abdien, H.M., Dessouki, A.A. )2010(. Prevalence of bacteria and parasites in White Ibis in Egypt. Vet. Ital., 46(3); 277-86.
3. Byrne, L., Vanstone, G.L., Perry, N.T., Launders, N., Adak, G.K., Godbole, G., et al. )2014(. Epidemiology and microbiology of Shiga toxin-producing Escherichia coli other than serogroup O157 in England, 2009–2013. J. Med. Mic., 63(9); 1181-1188.
4. Croxen, M.A., Law, R.J., Scholz, R., Keeney, K.M., Wlodarska, M., Finlay, B.B. )2013(. Recent advances in understanding enteric pathogenic Escherichia coli. Clin. Mic. Rev., 26(4); 822-880.
5. Estrada-García, T., Cerna, J.F., Paheco-Gil, L., Velázquez, R.F., Ochoa, T.J., Torres, J., et al. )2005(. Drug-resistant diarrheogenic Escherichia coli, Mexico. Emerg. Infect. Dis., 11(8); 1306.
6. Fagan, P.K., Hornitzky, M.A., Bettelheim, K.A., Djordjevic, S.P. )1999(. Detection of Shiga-like toxin (stx1 andstx2), intimin (eaeA), and enterohemorrhagic Escherichia coli (EHEC) hemolysin (EHEC hlyA) genes in animal feces by multiplex PCR. Appl. Environ. Microbiol., 65(2); 868-872.
7. Friesema, I., Van Der Zwaluw, K., Schuurman, T., Kooistra-Smid, M., Franz, E., Van Duynhoven, Y., et al. )2014(. Emergence of Escherichia coli encoding Shiga toxin 2f in human Shiga toxin-producing coli (STEC) infections in the Netherlands, January 2008 to December 2011. Eurosurveillance., 19(17(; 20787.
8. Gholami-Ahangaran, M., Zia-Jahromi, N. )2014(. Identification of shiga toxin and intimin genes in Escherichia coli detected from canary (Serinus canaria domestica). Toxicol.Indust. Health., 30(8); 724-727.
9. Gioia-Di Chiacchio, R.M., Cunha, M.P.V., Sturn, R.M., Moreno, L.Z., Moreno, A.M., Pereira, C.B.P., et al. )2016(. Shiga toxin-producing Escherichia coli (STEC): Zoonotic risks associated with psittacine pet birds in home environments. Vet. Microbiol., 184; 27-30.
10. Guenther, S., Ewers, C., Wieler, L.H. )2011(. Extended-spectrum beta-lactamases producing coli in wildlife, yet another form of environmental pollution. Front. Microbiol., 2; 246.
11. Haag-Wackernagel, D., Moch, H. )2004(. Health hazards posed by feral pigeons. J Infect., 48(4); 307-313.
12. Hosseini, S., Ranjbar, R., Zahrayi Salehi, T. )2016(. The Identification of pathogenic Escherichia coli strains to carry intimin gene isolated from various water sources. Iran. J. Med. Microbiol., 10(4); 17-24.
13. Kimpe, A., Decostere, A., Martel, A., Haesebrouck, F., Devriese, L.A. )2002(. Prevalence of antimicrobial resistance among pigeon isolates of Streptococcus gallolyticus, Escherichia coli and Salmonella enterica serotype Typhimurium. Avian Pathol., 31(4); 393-397.
14. Mora, A., Blanco, M., Blanco, J.E., Dahbi, G., López, C., Justel, P., et al. )2007(. Serotypes, virulence genes and intimin types of Shiga toxin (verocytotoxin)-producing Escherichia coli isolates from minced beef in Lugo (Spain) from 1995 through 2003. BMC Microbiol., 7(1); 13.
15. Morabito, S., Dell’Omo, G., Agrimi, U., Schmidt, H., Karch, H., Cheasty, T., et al. )2001(. Detection and characterization of Shiga toxin-producing Escherichia coli in feral pigeons. Vet. Microbiol., 82(3); 275-283.
16. Nolan, L.K., Barnes, H.J., Vaillancourt, J.P., Abdul-Aziz, T., Logue, C.M. )2020(. Colibacillosis. In: Disease of Poultry. Swayne, D.E., Boulianne, M., Logue, C.M., McDougald, L.R., Nair, V. and Suarez, D.L. editors. 14th ed., Massachusetts: Wiley-Blackwell, pp: 770-790.
17. Paton, J.C., Paton, A.W. )1998(. Pathogenesis and diagnosis of Shiga toxin-producing Escherichia coli Clin. Microbiol. Rev., 11(3); 450-479.
18. Persad, A.K., Lejeune, J.T. )2015(. Animal reservoirs of Shiga toxin-producing Escherichia coli. In Enterohemorrhagic Escherichia coli and Other Shiga Toxin-Producing E. coli. Am. Soc. Microbiol., 231-244.
19. Sandvig, K. )2001(. Shiga toxins. Toxicon, 39(11); 1629-1635.
20. Schmidt, H., Scheef, J., Morabito, S., Caprioli, A., Wieler, L.H., Karch, H. )2000(. A new Shiga toxin 2 variant (Stx2f) from Escherichia coli isolated from pigeons. Appl. Environ. Microbiol., 66(3); 1205-1208.
21. Ziauddini, A.H., Gholami-Ahangaran, M., Ahmadi-Dastgerdi, A. )2020(. Detection of virulence genes of intimin, hemolysin and shigatoxin in Escherichia coli isolated from Pscittacin. New Cell. Molecul. Biotechnol. J., 10(38); 61-68.