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بررسی بی ضرری انتروکوک های پروبیوتیکی بومی جدا شده از محصولات شیری و شیر انسان با استفاده از روش های بیوشیمیایی و مولکولی | ||
| علوم دامی | ||
| مقاله 7، دوره 31، شماره 120، آذر 1397، صفحه 89-102 اصل مقاله (1.12 M) | ||
| نوع مقاله: مقاله پژوهشی | ||
| شناسه دیجیتال (DOI): 10.22092/asj.2017.114810.1498 | ||
| نویسندگان | ||
| مهزاد خطیبی1؛ نرگس واسجی2؛ بهزاد صالحی3؛ ناهید مژگانی* 4 | ||
| 1کارشناس ارشد میکروبیولوژی-دانشگاه آزاد اسلامی واحد کرج | ||
| 2عضو هیئت علمی موسسه تحقیقات علوم دامی کشور، سازمان تحقیقات،آموزش و کشاورزی،کرج | ||
| 3عضو هیئت علمی دانشگاه آزاد اسلامی واحد کرج | ||
| 4عضو هیئت علمی موسسه تحقیقات واکسن و سرم سازی رازی، سازمان تحقیقات،آموزش و کشاورزی،کرج | ||
| چکیده | ||
| هدف از این مطالعه، تعیین بی ضرری و ایمنی گونه های انتروکوک جدا شده از شیر بز، پنیر سنتی و نمونه های شیر مادر بود. نمونه های انتروکوکوس جدا شده به وسیله ویژگی های فنوتیپی و تعیین توالی 16srRNA تا سطح جنس شناسایی شدند و خواص پروبیوتیکی آن ها، زنده مانی در شرایط اسیدی و حضور در درصدهای مختلف نمک صفراوی در زمان های مختلف بررسی گردید. فعالیت ضد میکروبی این باکتری ها بر علیه باکتری های بیماری زا و مقاومت آنها نسبت به انواع آنتی بیوتیک ها نیز ارزیابی شد. جهت بررسی بی ضرری انتروکوکوک های شناسایی شده، حضور یا عدم حضور تعدادی از ژن های ویرولانس(asa1, hyl, esp, agg, gelE, cylA, cylB, cylM, efaAfm, efaAfs) و مقاومت به ونکومایسین (vanA, vanH, vanR , vanY ) با استفاده از روش های بیوشیمیایی و مولکولی سنجیده شد. مقاومت فنوتیپی انتروکوک های انتخاب شده در برابر لیپاز، DNase و همولیز گلبول های قرمز نیز تعیین گردید. نتایج این تحقیق خصوصیات پروبیوتیکی بعضی از این گونه ها را مشخص کرد. 4 جدایه تحمل اسید و نمک صفرا را نشان دادند و اثرات ضد میکروبی چشمگیری داشتند. تمامی سویه های انتروکوکوس فاسیوم جدا شده از شیر مادر فاقد ژن های ویرولانس بودند. در آزمایشات فنوتیپی ویرولانس در سویه ها، همه سویه ها از نظر فعالیت همولیزی آلفاهمولیتیک بودند. همه سویه ها به تتراسایکلین، سفالوتین، آموکسی سیلین، سفازولین و داکسی سایکلین حساسیت نشان دادند و همه آن ها به جز سویه TA00154، نسبت به نالیدیکسیک اسید و تری متوپریم سولفامتاکسازول، مقاوم بودند. | ||
| کلیدواژهها | ||
| پروبیوتیک؛ انتروکوکوس؛ 16srRNA؛ ژنهای ویرولانس؛ مقاومت ونکومایسین | ||
| عنوان مقاله [English] | ||
| Evaluating the Safety of Indigenous Enterococcus species isolated from dairy products and Breast milk by Biochemical and Molecular Methods | ||
| نویسندگان [English] | ||
| Mahzad Khatibi1؛ Narges Vaseji2؛ Behzad Salehi3؛ Nahid Mojgani4 | ||
| 1Azad university of Karaj-Iran | ||
| 2Animal Science Research Institute of Iran, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran. | ||
| 3Azad university of Karaj-Iran | ||
| 4Razi Vaccine and Serum Research Institute.Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran. | ||
| چکیده [English] | ||
| Enterococcus plays an important role in various industries. Some Enterococci isolated from dairy and breast milk has been reported as probiotics. However, these species selected as starter cultures or as probiotics should be evaluated for their safety and the absence of different virulence traits including virulence genes and antibiotic resistance. The aim of this study was to determine the safety of locally isolated Enterococcus species biochemically and targeting essential virulence genes by PCR method. A number of suspected Enterococcus species isolated from ewe milk, traditional cheese and mothers' milk were identified to genus level by phenotypic characteristics and 16SrRNA sequencing. The selected isolates were screened for their probiotic properties by determining their acid and bile resistance, antibacterial activity against a number of pathogens and antibiotic resistance. Phenotypic virulence parameters including lipase, DNAse and hemolysis of red blood cells, was determined. The presence or absence of a number of virulence genes including asa1, hyl, esp, agg, gelE, cylA, cylB, cylM, efaAfm, efaAfs and vancomycin resistance genes including vanA, vanH, vanR and vanY was evaluated. The results of this study determined the probiotic properties of some of these species. Four isolates showed acid and bile salt tolerance and showed significant antimicrobial effects. All isolates from Breast milk, lacked virulence genes. In virulance phenotypic experiments, all strains were alpha hemolytic. All strains were susceptible to Tetracycline, Cephalothin Amoxicillin, Cefazolin and Doxycycline and all of them except the TA00154 strain, were resistant to Nalidixic acid and Trimethoprim sulfamethoxazole. | ||
| کلیدواژهها [English] | ||
| Probiotic, enterococcus, 16S rRNA, virulence genes, vancomycin resistance | ||
| مراجع | ||
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