جداسازی و غربالگری باکتریوفاژهای دریایی به منظور کاربرد در پیشگیری از بروز بیماری ویبریوزیس در میگوی سفید غربی: DOR: 20.1001.1.17359880.1400.14.3.4.5
محورهای موضوعی : مجله پلاسما و نشانگرهای زیستیسمیرا زارعی 1 , نیما بهادر 2 , مریم میربخش 3 , محمد خلیل پذیر 4
1 - گروه میکروب شناسی، دانشکده علوم، کشاورزی و فناوری‌های نوین، واحد شیراز، دانشگاه آزاد اسلامی، شیراز
2 - گروه میکروب شناسی، دانشکده علوم، کشاورزی و فناوری‌های نوین، واحد شیراز، دانشگاه آزاد اسلامی، شیراز
3 - مؤسسه تحقیقات علوم شیلاتی کشور، سازمان تحقیقات، آموزش و ترویج کشاورزی، تهران، ایران
4 - پژوهشکده میگوی کشور، موسسه تحقیقات علوم شیلاتی کشور، سازمان تحقیقات، آموزش و ترویج کشاورزی، بوشهر، ایران
کلید واژه: میگوی سفید غربی, جداسازی و غربالگری, باکتریوفاژ, بیماری ویبریوزیس,
چکیده مقاله :
زمینه و هدف: یکی از بزرگ ترین تهدیداتی که صنعت آبزی‌پروری با آن مواجه است، بیماری‌های عفونی باکتریایی و استفاده از آنتی بیوتیک ها است. فاژدرمانی یک جایگزین قابل اعتماد برای آنتی‌بیوتیک‌ها برای غیرفعال‌کردن باکتری به عنوان عوامل اصلی بیماری‌زا در صنعت آبزی‌پروری می باشد. هدف از تحقیق حاضر غربالگری گونه های ویبریو در سواحل استان بوشهر و کنترل این عامل بیماری زای میگو با استفاده از فاژ بود.روش کار: جهت انجام این مطالعه آب در سال 1398 و از مناطق نزدیک به سواحل استان بوشهر نمونه برداری شد. باکتری های عامل ویبریوزیس با استفاده از آزمایش های بیوشیمیایی و توالی یابی مولکولی تعیین شد. انتخاب باکتری هدف با توجه به نتایج پروفایل آنتی بیوگرام باکتری های غالب و پاتوژن ها انجام گردید. جهت تعیین تیتر فاژ محیط کشت TCBS به کار برده شد و شمارش پلاگ ها 24 ساعت پس از گرمخانه گذاری در دمای 30 درجه سانتی گراد انجام گرفت.یافته ها: از آب های آلوده به فاضلاب بوشهر، 5 باکتری(Vibrio alginolyticus strain SeqID، Vibrio xuii strain Z-03Phage، Vibrio alginolyticus SZ/10 gene، Vibrio rotiferianus strain SRPR-Phage، Vibrio harveyi strain Z05) جداسازی شد. با توجه به نتایج تعیین پروفایل آنتی بیوگرام، قطر هاله ی عدم رشد و آزمون بیوشیمایی باکتری گونه های مورد مطالعه، باکتری Vibrio alginolyticus strain SeqID به عنوان گونه ی باکتری غالب انتخاب شد. بر اساس نتایج حاصل از آنالیز مولکولی 16srDNA سویه جداسازی شده باکتری با 100% شباهت با باکتری ویبریو آلجینولایتیکوس بود، براساس نتایج دامنه میزبانی، فاژ Vibrio alginolyticus strain Samira Phage قادر به تشکیل پلاگ بود.نتیجه گیری: باکتریوفاژ Vibrio alginolyticus strain Samira Phage جداسازی شده اثر باکتری کشی با دامنه ای وسیع در برابر باکتری های ویبریوی شناسایی شده از سواحل بوشهر داشت که نشان می دهد این باکتریوفاژ می تواند کاندیدی مناسبی برای استفاده در فاژ درمانی بر علیه بیماری ویبریوز باشد.
The aim of this study was to screen Vibrio species off the coast of Bushehr province and control this shrimp pathogen using phage.For this study, water was sampled in 1398 from areas close to the coast of Bushehr province. Bacteria that cause Vibriosis were determined using biochemical tests and molecular sequencing. Bacterial selection was performed according to the results of antibiogram profiles of dominant bacteria and pathogens. TCBS was used to determine the phage titer of the culture medium and the plugs were counted 24 hours after incubation at 30 ° C.5 bacteria (Vibrio alginolyticus strain SeqID, Vibrio xuii strain Z-03Phage, Vibrio alginolyticus SZ / 10 gene, Vibrio rotiferianus strain SRPR-Phage, Vibrio harveyi strain Z05 were isolated from Bushehr wastewater contaminated waters). Vibrio alginolyticus strain SeqID was selected as the dominant bacterial species according to the results of antibiogram profile determination, growth halo diameter and bacterial biochemical test of the studied species. Based on the results of 16srDNA molecular analysis, the isolated strain of the bacterium was 100% similar to Vibrio alginolyticus. Based on the results of the host domain, Vibrio alginolyticus strain Samira Phage was able to form plugs.The isolated bacteriophage Vibrio alginolyticus strain Samira Phage had a bactericidal effect with a wide range against Vibrio bacteria identified from the shores of Bushehr, which indicates that this bacteriophage can be a good candidate for use in phage therapy Be against viberosis.Keywords: Isolation and screening, Bacteriophage, Vibriosis, Western white shrimp
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