پروفایل ژنی تولید انتروتوکسین جدایه‌های استافیلوکوکوس اورئوس داری مقاومت چندگانه آنتی‌بیوتیکی در محصولات لبنی سنتی استان فارس

محورهای موضوعی : میکروب شناسی غذایی

مریم بیگی ¹ , محمد کارگر ²

1 - گروه میکروبیولوژی، واحد جهرم، دانشگاه آزاد اسلامی، جهرم، ایران
2 - گروه میکروبیولوژی، واحد جهرم، دانشگاه آزاد اسلامی، جهرم، ایران

تاریخ دریافت : 1399/05/27 تاریخ پذیرش : 1399/05/27 تاریخ انتشار : 1399/06/01

کلید واژه: استافیلوکوکوس اورئوس, محصولات لبنی, انتروتوکسیژنیک, مقاومت چندگانه آنتی بیوتیکی,

چکیده مقاله :

سابقه و هدف: استافیلوکوکوس اورئوس از مهمترین عوامل مسمومیت غذایی در جهان است. هدف از این پژوهش ارزیابی فراوانی ژن‌های انتروتوکسینو شناسایی جدایه‌های دارای مقاومت چند دارویی در محصولات لبنی سنتی استان فارس می‌باشد.مواد و روش‌ها: این پژوهش به صورت مقطعی_توصیفی بر روی 300 نمونه محصولات لبنی سنتی(پنیر، شیر، کشک، دوغ و سرشیر) انجام گرفت. شناسایی و تائید جدایه‌های استافیلوکوکوس اورئوس با استفاده از آزمون‌های استاندارد بیوشیمیایی و تکثیرژن‌هایnucو 23S rRNA با روش PCR انجام شد. سپس به منظور ارزیابی ژن‌های انتروتوکسین از روشPCRچندگانه استفاده شد. همچنین حساسیت جدایه‌ها برای 16 آنتی‌بیوتیک با روش دیسک دیفیوژن مطابق معیار CLSI ارزیابی گردید.یافته‌ها: از مجموع نمونه‌های مورد بررسی، 120 جدایه (40%)‌ استافیلوکوکوس اورئوس تشخیص داده شدند. بیشترینفراوانیآلودگی مربوط به پنیر 38 نمونه (66/31%)و کمترین آن مربوط به دوغ 10 نمونه (33/8%) بود. 91 نمونه (83/75%) از جدایه‌های آلوده حاوی ژن انتروتوکسین بودند. بیشترین فراوانی ژن‌های انتروتوکسین، مربوط به sea (17/54%) و به دنبال آن به ترتیب seb ، seg ، see، sedو sec با فراوانی50%، 33/28%، 67/16%، 17/4% و 50/3% بود. همچنین از 64 سویه استافیلوکوکوس اورئوس (33/56%) دارای مقاومت چند دارویی، 54 جدایه (38/84%) واجد ژن های انتروتوکسین بودند.نتیجه‌گیری: نتایج پژوهش نشان دهنده فراوانی قابل توجه جدایه‌های استافیلوکوکوس اورئوس دارای مقاومت چند دارویی مولد انتروتوکسین در منطقه مورد بررسی بود. از این‌رو پایش مداوم مولکولی جدایه‌های استافیلوکوکوس اورئوس و همچنین مقایسه پروفایل ژنتیکی جدایه‌ها با نمونه‌های کلینیکی پیشنهاد می‌گردد.

چکیده انگلیسی:

Background & Objectives: Staphylococcus aureus is one of the most importantfactors of food poisoning in the world. The objective of this research is to investigate the frequency amount of enterotoxin genes in traditional dairy products with multidrug resistance isolates in Fars province.  Materials and Methods: This cross-sectional descriptive study was performed on 300 samples of traditional dairy products.  In order to identify Staphylococcus aureus isolates in addition to standard biochemical tests, according to PCR method, nuc and 23S rRNA genes proliferation were confirmed. Enterotoxin genes were identified by using of the multiple PCR method. The susceptibility of the isolates for 16 antibiotics was investigated by using of the disk diffusion method according to CLSI criteria.   Results: 120 isolates (40%) were identified as Staphylococcus aureus. The highest frequency of contamination is related to Cheese with 38 samples (31.66%) and its lowest relates to Doogh with 10 samples (8.33%). 91 samples (75.83%) of contaminated isolates were included of the enterotoxin gene, the highest frequency of enterotoxin gene was related to sea (54.17%) and its following, seb, seg, see, sed and sec respectively were included the frequency 50%, 33.33%. 28%, 16.67%, 4.17% and 3.50%. Also out of 64 strains of Staphylococcus aureus (56.33%) with multidrug resistance, 54 isolates (84.38%) had enterotoxin genes.  Conclusion: The finding indicated that a significant frequency of Staphylococcus aureus isolates with enterotoxin producing multidrug resistance. Therefore, continuous molecular monitoring of Staphylococcus aureus isolates and also evaluation of the association of genetic profiles with clinical specimens is recommended.

منابع و مأخذ:

1. Eslami M, Koohi MK, adehashem E, Khadiri B, Keshavarz H. Survey the Presence of
   Coagulase Positive Staphylococcus aureus in Cottage Cheeses Produced from Sheep Milk and
   Sold in Marand County. J Food Sci Tech. 2015; 12(46): 211-218.
   2. Alizadeh S, Amini K. Determining the Presence of irulence Genes Panton alentine
   Leukocidin Pvl and Methicillin Resistance Gene Meca in Staphylococcus aureus Strains
   Isolated from Food Samples by Multiplex PCR and Antibiotic Resistance. J Rafsanjan Univ
   Med Sci. 2015; 14(5): 27-34[In persian].
   3. Mahdavi S. Prevalence of Six Genes Encoding Enterotoxins Production of Staphylococcus
   aureus Isolated from White-Brined-Cheese by Multiplex PCR. J Food Hygiene. 2014; 3(4):
   1-7.
   4. Tavakoli H R, jodayi A A, Imani Foladi A A, sarshar M, Rafati H, Asadi bagh asiab B.
   Comman Types Of Staphylococcus Aureus Enterotoxin In Meaty Foods. Iran J Clin Infect Dis
   Trop Med. 2013; 17(59): 9-15.
   5. Podkowik M, Park J, Seo K, Bystroń J, Bania J. Enterotoxigenic potential of
   coagulase-negative staphylococci. International. J food microbiol. 2013; 163: 34-40.
   6. Luss JP, antonder I. The occurrence of indicator bacteria on hands and aprons of food
   handlers in the delicatessen sections of a retail group. J Food Control. 2007; 18(4): 326-332.
   7. Ahmadi , Tajbakhsh E, Momtaz H. Detection of the Antibiotic Resistance Pattern in
   Staphylococcus aureus Isolated from Clinical Samples Obtained from Patients Hospitalised in
   Imam Reza Hospital, Kermanshah. J Microbial World. 2014; 6(4): 209-311.
   8. Magiorakos A. Srinivasan A, Carey R, Carmeli Y, Falagas M, Giske C, Olsson‐Liljequist B.
   Multidrug‐resistant, extensively drug‐resistant and pandrug‐resistant bacteria: an international
   expert proposal for interim standard definitions for acquired resistance. Clin microbiol infect.
   2012; 18(3): 268-281.
   9. Mirzaei H, Javadi A, Farajli M, Shah-Mohammadi A, Monadi A, Barzegar A. Prevalence of
   Staphylococcus aureus resistant to methicillin in traditional cheese and cream: a study in city
   of Tabriz, Iran. J et Res. 2012; 1.70-65 :)76(
   10. Brown DFJ, Edwards DI, Hawkey PM, Morrison D, Ridgway GL, Towner KJ, Wren MW.
   Guidelines for the laboratory diagnosis and susceptibility testing of Methicillin-Resistant
   Staphylococcus aureus (MRSA). J Antimicrob Chemother. 2005; 56: 1000-1018.
   11. Mollaei M, Rashki A. The Prevalence of Adhesive Surface Encoding Genes in
   Staphylococcus aureus Isolated from Hospitalized Patients in abol-Iran by Multiplex PCR. J
   Fasa Univ Med Sci. 2016; 6(3): 296-302.
   12. Chapaval L, Moon D, Gomes J, Duarte F, Tsai S. Use of PCR to Detect Classical
   Enterotoxins Genes and Toxic Shock Syndrom Toxin-1 Gene (tst) in Staphylococcus aureus
   Isolated from Crude Milk and Determination of Toxin Productivites of S.aureus. Arq Inst Biol,
   Sao Paulo. 2006; 73(2): 165-169.
   13. Cremonesi P, Luzzana M, Brasca M, Morandi S, Lodi R, imercati C, Agnellini D,
   Caramenti G, Moroni P, Castiglioni B. Development of a multiplex PCR assay for the
   identification of Staphylococcus aureus enterotoxigenic strains isolated from milk and dairy
   products. Mol Cell Probes. 2005; 19: 299-305.
   14. Mahdiyoun, S M, Ahanjan M, Goudarzi M, Rezaee R. Prevalence of Antibiotic Resistance in
   Methicillin-Resistant Staphylococcus aureus and Determining Aminoglycoside Resistance
   Gene by PCR in Sari and Tehran Hospitals. J Mazandaran Univ Med Sci. 2015; 25(128):
   97-107 [In persian].
   15. Nazari R, Godarzi H, Rahimi Baghi F, Moeinrad M. Enterotoxin Gene Profiles among
   Staphylococcus aureus Isolated from Raw Milk. Iran J et Res. 2014; 15(4): 409.
   16. Alni, R, Mohammadzadeh A, Mahmoodi P, Alikhani M. Detection of Toxic Shock Syndrome
   Toxin (tsst) Gene Among Staphylococcus aureus Isolated from Patients and Healthy Carriers.
   Avicenna J Clin Microbiol Infect. 2018; 5(1): 1-5.
   17. MahdaviS. Prevalence of Six Genes Encoding Enterotoxins Production of Staphylococcus
   aureus Isolated from White-Brined-Cheese by Multiplex PCR. Journal of Food Hygiene. 2014;
   3(4): 1-7.
   18. CLSI, Performance Standards for Antimicrobial Susceptibility Testing 30th ed. CLSI
   Supplement M100, CLSI, Wayne, Pa, USA; 2020.
   19. Ghazavi N, Rahimi E, Esfandiari , Shakerian A. Enterotoxigencity, distribution of
   enterotoxigenic genes and antibiotic resistance pattern of Staphylococcus aureus isilated from
   traditional sweet. Tropical Biomedicine. 2018; 35(4): 880-892.
   20. Shekarforoush SS, N Rokni, G Karim, Rohani Razavi SM, Kiaie S M M, Abbasvali M. Study
   on the Overview on Food Borne Bacteria in Foodstuffs with Animal Origin in Iran, Part Two:
   Meat and Meat Products. J Food Hyg. 2012; 2(3): 1-14.
   21. Sadeghi, M R. The prevalence of Enterotoxigenic Isolates of Staphylococcus aureus in Bovine and Sheep Bulk Tank Milk Samples of Maku. J et Microbiol. 2015; 11(1): 27-38.
   22. Imani-Fooladi A.A, Riazipour M, Sattari M. Molecular and serological detection of
   Enterotoxigenic Staphylococcus aureus from traditionally dairy products. J Shahrekord Univ
   Med Sci. 2010; 11(4): 19–26. [persian]
   23. Singh P, Prakash A. Prevalance of coagulase positive pathogenic Staphylococcus aureus in
   milk and milk products collected from unorganized sector of Agra. Journal Guide-Acta
   Agriculturae Slovenica. 2010; 96(1): 37–41.
   
   24. Mohammadi Sani A. The survey of the distributed cheese to listeria monocytogen,
   Staphylococcus aureus and Ecoli pathogens in Mashhad and Ghochan. Twelf th national
   congress of Iran environment health, Shahid Beheshti medical sciences university. 522-533.
   25. Molla Abaszadeh H, Haji Sheikhzadeh B. Surveying the contamination rate, sensibility and
   antimicrobial resistance patterns in Staphylococcus aureus isolated from Traditional cheese
   consumed in Qotur of Khoy province. J Fasa univ med sci. 2014; 4(2): 209-217.
   26. Abbasi S, Taei S, amanzad B. The prevalence of methicillin-resistant Staphylococcus aureus
   strains producing enterotoxin A and B. Tehran Univ Med J. 2016; 73(11):778-83 [In persian].
   27. Morandi S, Brasca M, Lodi R, Cremonesi P, Castiglioni B. Detection of classical enterotoxins
   and identification of enterotoxin genes in Staphylococcus aureus from milk and dairy products.
   et microbiol. 2007; 124: 66-72.
   28. icosa GN, Le Loir A, Le Loir Y, de Carvalho AF, Nero LA. egc characterization of
   enterotoxigenic Staphylococcus aureus isolates obtained from raw milk and cheese. Int J Food
   Microbiol. 2013; 165: 227-30.
   29. Mashouf R, Hosseini S.M, Mousavi S.M. Arabestani M. Prevalence of Enterotoxin Genes and
   Antibacterial Susceptibility Pattern of Staphylococcus aureus Strains Isolated from Animal
   Originated Foods in West of Iran. Oman med j. 2015; 30 (4): 283.
   30. Adwan G, Abu-Shanab B, Adwan K. Enterotoxigenic Staphylococcus aureus in raw milk in
   the North of Palestine. Turk J Biol. 2005; 29: 229-232.
   31. Ahmadi E, Ghuzivandi A, Ebrahim Pourbaser, K, Karimi Dareabi, H. Identification of sea and
   seb frequency in Staphylococcus aureus isolated from cow, sheep and goat mastitic milk
   samples in Sanandaj city. Iran et J. 2018; 14(2): 5-13.
   32. Imani Fooladi A.A, Tavakoli H.R, Naderi A. Detection of enterotoxigenic Staphylococcus
   aureus isolates in domestic dairy products. Iran J Microbiol. 2010; 2(3): 137–142.
   33. Khazaei S, Pourtahmaseby P, Kanani M, Madani S.H, Malekianzadeh E. Staphylococcus
   aureus Resistance to ancomycin: A Six Years Survey,(2006-2012). Med J Tabriz Univ Med
   Sci Health Services. 2013; 35(5): 40-45.
   34. Ma Y, hao Y, Tang J, Tang C, Chen J, Liu J. Antimicrobial susceptibility and presence of
   resistance & enterotoxins/enterotoxin-likes genes in Staphylococcus aureus from food. CyTA-J
   Food. 2018; 16(1): 76-84.
   35. huge S.Y, Tan D.M, Li .G. Research on Staphylococcus aureus enterotoxin property and
   drug resistance in quick frozen flour product in Guangxi. Mod Prev Med. 2015; 42: 1765-1767.
   36. Teyhoo M, Mobin M, Mozafari NA, Moadab SR, Sedigh Bayan KH, Mones Rast SH. The
   prevalence of Toxic shock syndrome toxin -1 (TSST-1) producing clinical isolates of
   Staphylococcus aureus strain isolated from shohada hospital in Tabriz, Iran. Med lab j. 2011; 5
   (1): 38-44.[persian]
   37. Kumar R, Yadav B, Singh R. Antibiotic Resistance and Pathogenicity Factors in
   Staphylococcus aureus Isolated from Mastitic Sahiwal Cattle. J of biosciences. 2011; 36(1):
   175-88.
   38. Parastan R, Kargar M, Solhjoo K,Kafilzadeh F. Staphylococcus aureus biofilms: Structures,
   antibiotic resistance, inhibition, and vaccines. Gene Reports.2020; 20, 100739.
   39. Parastan R, Kargar M, Solhjoo K,Kafilzadeh F. A synergistic association between
   adhesion-related genes and multidrug resistance patterns of Staphylococcus aureus isolates
   from different patients and healthy individuals. Journal of Global Antimicrobial
   Resistance.2020; 22: 379-385.
   40. Moezi P, Kargar M, Doosti A, Khoshneviszadeh M. Multiplex touchdown PCR assay to
   enhance specificity and sensitivity for concurrent detection of four foodbornepathogens in raw
   milk. Journal of Applied Microbiology.2019; 127:262-273.
   41. Moezi P, Kargar M, Doosti A, Khoshneviszadeh M. Developing a multiplex real-time PCR
   with anew pre-enrichment to simultaneously detect four foodborne bacteria in milk. Future
   Microbiol. 201914(10):885–898.
   42. Dahyot S, Lbeurre J, Argemi , Francois P, Lemee L, Prevost G, et al. Multiple-Locus
   ariable Number Tandem Repeat Analysis (ML A) and Tandem Repeat Sequence Typing
   (TRST), helpful tools for subtyping Staphylococcus lugdunensis. Sci Rep. 2018; 8: 1-11.