دسته‌بندی ژنتیکی ایزولههای استافیلوکوکوس اورئوس جدا شده از گوشت مرغ بر پایه ژن Spa
محورهای موضوعی :
میکروب شناسی
حسن ممتاز
1
(استاد، گروه میکروبیولوژی، واحد شهرکرد، دانشگاه آزاد اسلامی، شهرکرد، ایران)
پریسا حیدری
2
(کارشناسی ارشد، گروه میکروبیولوژی، واحد شهرکرد، دانشگاه آزاد اسلامی، شهرکرد، ایران)
تاریخ دریافت : 1400/02/21
تاریخ پذیرش : 1401/05/06
تاریخ انتشار : 1400/10/01
کلید واژه:
ژن Spa,
ژنوتایپیینگ,
استافیلوکوکوس اورئوس,
گوشت مرغ,
استان چهارمحال و بختیاری,
چکیده مقاله :
هدف: استافیلوکوکوس اورئوس یکی از شایع‌ترین عوامل مسمومیت غذایی در انسان می‌‌باشد. روش‌‌های مختلفی از جمله ژنوتایپینگ بر اساس پروتئین A (Spa typing) و روش‌‌های مبتنی بر PCR جهت دسته بندی ژنتیکی این باکتری به کار رفته است. در مطالعه حاضر از ردیابی ژن Spa جهت ژنوتایپینگ ایزولههای استافیلوکوکوس اورئوس استفاده شد.مواد و روش‌ها: تعداد 100 نمونه گوشت خام مرغ از مراکز عرضه‌کننده گوشت مرغ در سطح بازار استان چهارمحال و بختیاری، اخذ شد. نمونه‌ها جهت جستجوی باکتری استافیلوکوکوس اورئوس به روش کشت میکروبی و مولکولی آزمایش و حضور ژن Spa در ایزولههای جدا شده به روش PCR و هضم آنزیمی جستجو شد.نتایج: از مجموع 23 ایزوله استافیلوکوکوس اورئوس مورد مطالعه، تعداد 6 ایزوله فاقد ژن Spa بودند. در بقیه ایزوله‌ها، قطعه‌ای با اندازه حدوداً 1100 تا 1500 جفت بازی ردیابی شد که بر اساس قطعه ژنی ردیابی شده، 17 ایزوله واجد ژن Spa در چهار ژنوتیپ I تا IV تقسیم‌بندی شدند؛ به طوری که 9 ایزوله در ژنوتیپ SpaI، 3 ایزوله در ژنوتیپ II Spa، 3 ایزوله در ژنوتیپ SpaIIIو 2 ایزوله در ژنوتیپ Spa IV قرار گرفتند.نتیجه‌گیری: وجود تنوع ژنومی بالا در این ایزوله‌ها نشانگر انتقال متقاطع آلودگی بوده و بنابراین، می‌‌توان با اجرای استانداردهای کنترل کیفیت و امنیت مواد غذایی در حین فرآیند تولید از حضور و رشد این باکتری در مواد غذایی جلوگیری کرد.
چکیده انگلیسی:
Objectives: Staphylococcus aureus is one of the most common causes of food poisoning in humans. Various methods including genotyping based on protein A (Spa typing) and PCR-based methods have been used for genetic classification of this bacterium. In the present study, Spa gene tracking was used for genotyping of Staphylococcus aureus isolates.Methods: 100 samples of raw chicken meat were collected from chicken meat supply centers in the market of Chaharmahal and Bakhtiari province. Samples were tested for Staphylococcus aureus bacteria by microbial culture and molecular methods and the presence of Spa gene in strains detected by PCR and enzymatic digestion.Results: Out of the total of 23 isolates of Staphylococcus aureus studied, 6 isolates lacked the Spa gene. In the rest of the isolates, a fragment with a size of about 1100 to 1500 bp was detected, and based on the detected gene fragment, 17 isolates containing the Spa gene were divided into four genotypes I to IV; So that 9 isolates were in SpaI genotype, 3 isolates in SpaII genotype, 3 isolates in SpaIII genotype and 2 isolates in SpaIV genotype.Conclusion: The presence of high genomic diversity in these isolates indicates cross-contamination of contamination and therefore can be prevented from the presence and growth of this bacterium in food by implementing quality control and food safety standards during the production process.
منابع و مأخذ:
Kadariya J, Smith TC & Thapaliya D. Staphylococcus aureus and staphylococcal food-borne disease: an ongoing challenge in public health. BioMed Research International. 2014; 827965. DOI:1155/2014/827965
Wei-Wei L, Zhu J, Zhen S, Liang X, Jiang Y & Ning L. Analysis of foodborne disease outbreaks in China mainland in 2011. Chines Journal of Food Hygiene. 2018; 30: 283-8.
Jackson CR, Davis JA & Barrett JB. Prevalence and characterization of methicillin-resistant Staphylococcus aureus isolates from retail meat and humans in Georgia. Journal of Clinical Microbiology. 2013; 51(4): 1199-207.
McCallum N, Berger-Bächi B & Senn MM. Regulation of antibiotic resistance in Staphylococcus aureus. International Journal of Medical Microbiology. 2010; 300(2-3): 118-29.
Papadopoulos P, Papadopoulos T, Angelidis AS, Boukouvala E, Zdragas A, Papa A, Hadjichristodoulou C & Sergelidis D. Prevalence of Staphylococcus aureus and of methicillin-resistant S. aureus (MRSA) along the production chain of dairy products in north-western Greece. Food Microbiology. 2018; 69: 43-50.
Wu S, Huang J, Wu Q, Zhang J, Zhang F, Yang X, Wu H, Zeng H, Chen M, Ding Y & Wang J. Staphylococcus aureus isolated from retail meat and meat products in China: incidence, antibiotic resistance and genetic diversity. Frontiers in Microbiology. 2018;
9: 2767.
Koreen L, Ramaswamy SV, Graviss EA, Naidich S, Musser JM & Kreiswirth BN. Spa typing method for discriminating among Staphylococcus aureus isolates: implications for use of a single marker to detect genetic micro-and macrovariation. Journal of Clinical Microbiology. 2004; 42(2): 792-9.
Strommenger B, Braulke C, Heuck D, Schmidt C, Pasemann B, Nubel U & Witte W. Spa typing of Staphylococcus aureus as a frontline tool in epidemiological typing. Journal of Clinical Microbiology. 2008; 46(2): 574-81.
Farahmand S, Haeili M, Darban-Sarokhalil D. Molecular Typing and Drug Resistance Patterns of Staphylococcus aureus Isolated from Raw Beef and Chicken Meat Samples. Iranian Journal of Medical Microbiology. 2020; 14(5): 478-89.
Kumar R, Yadav BR & Singh RS. Genetic determinants of antibiotic resistance in Staphylococcus aureus isolates from milk of mastitic crossbred cattle. Current Microbiology. 2010; 60(5): 379-86.
Afrough P, Pourmand MR, Zeinalinia N, Yousefi M, Abdossamadi Z & Bagherzadeh Yazdchi S. Molecular typing of clinical and nasal carriage isolates of Staphylococcus aureus by spa gene patterns. Journal of Mazandaran University of Medical Sciences. 2012; 22(94): 28-34.
Börjesson S. Antibiotic Resistance in Wastewater: Methicillin-resistant Staphylococcus aureus (MRSA) and antibiotic resistance genes. Doctoral dissertation, Linköping University Electronic Press, 2009.
Momtaz H, Dehkordi FS, Rahimi E, Asgarifar A & Momeni M. Virulence genes and antimicrobial resistance profiles of Staphylococcus aureus isolated from chicken meat in Isfahan province, Iran. Journal of Applied Poultry Research. 2013; 22(4): 913-21.
Omoe K, Ishikawa M, Shimoda Y, Hu DL, Ueda S & Shinagawa K. Detection of seg, seh, and sei genes in Staphylococcus aureus isolates and determination of the enterotoxin productivities of S. aureus isolates harboring seg, seh, or sei genes. Journal of Clinical Microbiology. 2002; 40(3): 857-62.
Soriano JM, Rico H, Molto JC & Manes J. Effect of introduction of HACCP on the microbiological quality of some restaurant meals. Food Control. 2002; 13(4-5): 253-61.
Huong BT, Mahmud ZH, Neogi SB, Kassu A, Van Nhien N, Mohammad A, Yamato
M, Ota F, Lam NT, Dao HT & Khan NC. Toxigenicity and genetic diversity of Staphylococcus aureus isolated from Vietnamese ready-to-eat foods. Food Control. 2010; 21(2): 166-71.
Gencay YE, Ayaz ND & Doğru AK. Enterotoxin gene profiles of Staphylococcus aureus and other Staphylococcal isolates from various foods and food ingredients. Erciyes Üniversitesi Veteriner Fakültesi Dergisi. 2010; 7(2): 75-80.
Eshraghi S & Salehipur Z. Detection of tst, sec, sea and sec +sea in staphylococcus aureus isolated from different food sampling. Tehran Medical University Journal. 2008; 67(7): 470-6.
Javadi A & Safarmashaei S. Microbial profile of marketed broiler meat. Middle-East Journal of Scientific Research. 2011; 9(5): 652-6.
Feizi A, Nazeri M & Pilevar A. Isolation of Staphylococcus spp. genera from broiler breeder flocks in East Azerbaijan Province of Iran: Prevalence and antimicrobial susceptibility. African Journal of Microbiology Research. 2012; 6(28): 5819-23.
Ajello L, Hay R, Topley WW & Wilson SG. Topley and Wilson's Microbiology and Microbial Infections: Medical Mycology. Arnold; 1998.
Songer JG & Post KW. Veterinary microbiology-E-book: bacterial and fungal agents of animal disease. Elsevier Health Sciences; 2004.
Salasia SI, Khusnan Z, Lammler C & Zschock M. Comparative studies on pheno-and genotypic properties of Staphylococcus aureus isolated from bovine subclinical mastitis in central Java in Indonesia and Hesse in Germany. Journal of Veterinary Science. 2004; 5(2): 103-9.
Bystroń JA, Bania J, Lis EL, Molenda J & Bednarski M. Characterisation of Staphylococcus aureus strains isolated from cows' milk. Bulletin of the Veterinary Institute in Puławy. 2009; 53(1): 59-63.
Karahan M, Acik MN & Cetinkaya B. Investigation of virulence genes by PCR in Stapylococcus aureus isolates originated from subclinical bovine mastitis in Turkey. Pakistan Veterinary Journal. 2011; 31(3): 249-53.
Bhati T, Nathawat P, Sharma SK, Yadav R, Bishnoi J & Kataria AK. Polymorphism in spa gene of Staphylococcus aureus from bovine subclinical mastitis. Veterinary World. 2016; 9(4): 421.
Frenay HM, Theelen JP, Schouls LM, Vandenbroucke-Grauls CM, Verhoef J, Van Leeuwen WJ & Mooi FR. Discrimination of epidemic and nonepidemic methicillin-resistant Staphylococcus aureus strains on the basis of protein A gene polymorphism. Journal of Clinical Microbiology. 1994; 32(3): 846-7.
Kuzma K, Malinowski ED, Lassa HE & Klossowska A. Analysis of protein A gene polymorphism in Staphylococcus aureus isolates from bovine mastitis. Bulletin of the Veterinary Instit27ute in Pulawy. 2005; 49: 41-4.
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Kadariya J, Smith TC & Thapaliya D. Staphylococcus aureus and staphylococcal food-borne disease: an ongoing challenge in public health. BioMed Research International. 2014; 827965. DOI:1155/2014/827965
Wei-Wei L, Zhu J, Zhen S, Liang X, Jiang Y & Ning L. Analysis of foodborne disease outbreaks in China mainland in 2011. Chines Journal of Food Hygiene. 2018; 30: 283-8.
Jackson CR, Davis JA & Barrett JB. Prevalence and characterization of methicillin-resistant Staphylococcus aureus isolates from retail meat and humans in Georgia. Journal of Clinical Microbiology. 2013; 51(4): 1199-207.
McCallum N, Berger-Bächi B & Senn MM. Regulation of antibiotic resistance in Staphylococcus aureus. International Journal of Medical Microbiology. 2010; 300(2-3): 118-29.
Papadopoulos P, Papadopoulos T, Angelidis AS, Boukouvala E, Zdragas A, Papa A, Hadjichristodoulou C & Sergelidis D. Prevalence of Staphylococcus aureus and of methicillin-resistant S. aureus (MRSA) along the production chain of dairy products in north-western Greece. Food Microbiology. 2018; 69: 43-50.
Wu S, Huang J, Wu Q, Zhang J, Zhang F, Yang X, Wu H, Zeng H, Chen M, Ding Y & Wang J. Staphylococcus aureus isolated from retail meat and meat products in China: incidence, antibiotic resistance and genetic diversity. Frontiers in Microbiology. 2018;
9: 2767.
Koreen L, Ramaswamy SV, Graviss EA, Naidich S, Musser JM & Kreiswirth BN. Spa typing method for discriminating among Staphylococcus aureus isolates: implications for use of a single marker to detect genetic micro-and macrovariation. Journal of Clinical Microbiology. 2004; 42(2): 792-9.
Strommenger B, Braulke C, Heuck D, Schmidt C, Pasemann B, Nubel U & Witte W. Spa typing of Staphylococcus aureus as a frontline tool in epidemiological typing. Journal of Clinical Microbiology. 2008; 46(2): 574-81.
Farahmand S, Haeili M, Darban-Sarokhalil D. Molecular Typing and Drug Resistance Patterns of Staphylococcus aureus Isolated from Raw Beef and Chicken Meat Samples. Iranian Journal of Medical Microbiology. 2020; 14(5): 478-89.
Kumar R, Yadav BR & Singh RS. Genetic determinants of antibiotic resistance in Staphylococcus aureus isolates from milk of mastitic crossbred cattle. Current Microbiology. 2010; 60(5): 379-86.
Afrough P, Pourmand MR, Zeinalinia N, Yousefi M, Abdossamadi Z & Bagherzadeh Yazdchi S. Molecular typing of clinical and nasal carriage isolates of Staphylococcus aureus by spa gene patterns. Journal of Mazandaran University of Medical Sciences. 2012; 22(94): 28-34.
Börjesson S. Antibiotic Resistance in Wastewater: Methicillin-resistant Staphylococcus aureus (MRSA) and antibiotic resistance genes. Doctoral dissertation, Linköping University Electronic Press, 2009.
Momtaz H, Dehkordi FS, Rahimi E, Asgarifar A & Momeni M. Virulence genes and antimicrobial resistance profiles of Staphylococcus aureus isolated from chicken meat in Isfahan province, Iran. Journal of Applied Poultry Research. 2013; 22(4): 913-21.
Omoe K, Ishikawa M, Shimoda Y, Hu DL, Ueda S & Shinagawa K. Detection of seg, seh, and sei genes in Staphylococcus aureus isolates and determination of the enterotoxin productivities of S. aureus isolates harboring seg, seh, or sei genes. Journal of Clinical Microbiology. 2002; 40(3): 857-62.
Soriano JM, Rico H, Molto JC & Manes J. Effect of introduction of HACCP on the microbiological quality of some restaurant meals. Food Control. 2002; 13(4-5): 253-61.
Huong BT, Mahmud ZH, Neogi SB, Kassu A, Van Nhien N, Mohammad A, Yamato
M, Ota F, Lam NT, Dao HT & Khan NC. Toxigenicity and genetic diversity of Staphylococcus aureus isolated from Vietnamese ready-to-eat foods. Food Control. 2010; 21(2): 166-71.
Gencay YE, Ayaz ND & Doğru AK. Enterotoxin gene profiles of Staphylococcus aureus and other Staphylococcal isolates from various foods and food ingredients. Erciyes Üniversitesi Veteriner Fakültesi Dergisi. 2010; 7(2): 75-80.
Eshraghi S & Salehipur Z. Detection of tst, sec, sea and sec +sea in staphylococcus aureus isolated from different food sampling. Tehran Medical University Journal. 2008; 67(7): 470-6.
Javadi A & Safarmashaei S. Microbial profile of marketed broiler meat. Middle-East Journal of Scientific Research. 2011; 9(5): 652-6.
Feizi A, Nazeri M & Pilevar A. Isolation of Staphylococcus spp. genera from broiler breeder flocks in East Azerbaijan Province of Iran: Prevalence and antimicrobial susceptibility. African Journal of Microbiology Research. 2012; 6(28): 5819-23.
Ajello L, Hay R, Topley WW & Wilson SG. Topley and Wilson's Microbiology and Microbial Infections: Medical Mycology. Arnold; 1998.
Songer JG & Post KW. Veterinary microbiology-E-book: bacterial and fungal agents of animal disease. Elsevier Health Sciences; 2004.
Salasia SI, Khusnan Z, Lammler C & Zschock M. Comparative studies on pheno-and genotypic properties of Staphylococcus aureus isolated from bovine subclinical mastitis in central Java in Indonesia and Hesse in Germany. Journal of Veterinary Science. 2004; 5(2): 103-9.
Bystroń JA, Bania J, Lis EL, Molenda J & Bednarski M. Characterisation of Staphylococcus aureus strains isolated from cows' milk. Bulletin of the Veterinary Institute in Puławy. 2009; 53(1): 59-63.
Karahan M, Acik MN & Cetinkaya B. Investigation of virulence genes by PCR in Stapylococcus aureus isolates originated from subclinical bovine mastitis in Turkey. Pakistan Veterinary Journal. 2011; 31(3): 249-53.
Bhati T, Nathawat P, Sharma SK, Yadav R, Bishnoi J & Kataria AK. Polymorphism in spa gene of Staphylococcus aureus from bovine subclinical mastitis. Veterinary World. 2016; 9(4): 421.
Frenay HM, Theelen JP, Schouls LM, Vandenbroucke-Grauls CM, Verhoef J, Van Leeuwen WJ & Mooi FR. Discrimination of epidemic and nonepidemic methicillin-resistant Staphylococcus aureus strains on the basis of protein A gene polymorphism. Journal of Clinical Microbiology. 1994; 32(3): 846-7.
Kuzma K, Malinowski ED, Lassa HE & Klossowska A. Analysis of protein A gene polymorphism in Staphylococcus aureus isolates from bovine mastitis. Bulletin of the Veterinary Instit27ute in Pulawy. 2005; 49: 41-4.
