بررسی تاثیر نانوامولسیون اسانس ترخون بر بیان ژن های حدت اشرشیاکلی انتروهموراژیک

نویسندگان
مریم عزیزخانی 1
فهیمه توریان 1
پولین شهره 2
راضیه پرتوی 2
محمدحسن شاهوی 3
1 دانشگاه تخصصی فن آوری های نوین آمل
2 گروه بهداشت مواد غذایی، دانشکده دامپزشکی، دانشگاه تخصصی فن آوری‌های نوین آمل
3 دانشگاه تخصصی فن آوری‌های نوین آمل
10.52547/fsct.17.106.121
چکیده
اشرشیاکلی O157:H7 انتروهموراژیک از مهمترین و شایع ترین پاتوژن­های غذایی در سراسر دنیاست که در حال کسب مقاومت در برابر برخی ترکیبات ضدمیکربی سنتزی رایج می­باشد. هدف از این پژوهش تعیین حداقل غلظت مهار کنندگی (MIC) و حداقل غلظت باکتری­کشی (MBC)نانو امولسیون اسانس ترخون (Artemisia dracunculus) برای سویه انتروهموراژیک اشرشیاکلی و سپس تاثیر غلظت­های تحت MIC آن بر نرخ رشد و بیان ژن­های حدت stx1) و (stx2 بود. نانوامولسیون اسانس ترخون به روش فراصوت تهیه و اندازه قطرات و پتانسیل زتای آن تعیین شد. MIC و MBC اسانس و نانوامولسیون با استفاده از روش میکرودایلوشن براث تعیین شد. میزان رشد و بیان ژن­های stx1A و stx2A در اشرشیاکلی پس از تیمار با غلظت­های مختلف تحت MIC بررسی شد. استراگول به عنوان اصلی­ ترین ماده تشکیل دهنده اسانس شناسایی شد. قطر ذرات نانوامولسیون به طور متوسط ​​50 نانومتر و پتانسیل زتا 30- میلی ولت بود. مقادیر MIC اسانس و نانوامولسیون، به ترتیب، 11/0± 58/0 و 07/0± 33/0میلی­گرم در میلی لیتر و MBC معادل، به ترتیب، 20/0± 65/0 و 15/0± 38/0میلی­گرم در میلی لیتر به دست آمد. نانوامولسیون نسبت به اسانس خالص دارای اثر مهارکنندگی بیشتری در برابر رشد باکتری بود. در پایان دوره 72 ساعته، تیمار با نانوامولسیون در غلظت 75 درصد MIC منجر به کاهش نسخه برداری از stx1A و stx2A به ترتیب برابر 75/3 و 10/4 برابر گردید در حالی که در غلظت 75 درصد MIC اسانس، میزان نسخه برداری از stx1 و stx2 در مقایسه با شاهد به ترتیب 91/1 و 02/2 برابر کاهش یافت. بیشتر بودن فعالیت مهارکنندگی نانوامولسیون اسانس ترخون در مقایسه با اسانس خالص در برابر رشد و تولید شیگاتوکسین اشرشیاکلی پتانسیل آن را برای کاربرد به عنوان نگهدارنده خوراکی طبیعی و نیز راه حلی جهت مشکل جهانی ظهور میکرب­های مقاوم به آنتی بیوتیک نشان می­دهد.
کلیدواژه‌ها
اسانس
اشرشیاکلی انتروهموراژیک
ترخون
شیگاتوکسین
نانوامولسیون

موضوعات

عنوان مقاله English

Evaluating the effect of Nanoemulsion of Artemisia dracunculus Essential Oil on Expression of virulence genes in enterohemorrhagic Escherichia coli

نویسندگان English

Maryam Azizkhani 1
Fahimeh Tooryan 1
P. shohreh 2
Razieh Partovi 2
MH Shahavi 3
1 Amol University of Special Modern Technologies
2 Amol University of Special modern Technologies
3 Amol University of Special modern Technologies
چکیده English

​Escherichia coli O157:H7 is one of the most important and common foodborne pathogens in the world which is being resistant against some current synthetic antimicrobials. The aim of this study was to determine the minimum inhibitory concentration (MIC) and the minimum bacterial concentration (MBC) of Artemisia dracunculus (tarragon) essential oil and its nanoemulsion for enterohemorrhagic strain of Escherichia coli and then the effect of sub-MIC concentrations on growth rate and gene expression of virulence genes (stx1A and stx2A). Nanoemulsion of tarragon essential oil was prepared by the ultrasound method and the droplet size and zeta potential were determined. MIC and MBC of essential oil and nanoemulsion were determined using the broth microdilution method. The growth rate and expression of stx1A and stx2A genes in Escherichia coli were assessed after treatment with different concentrations of sub-MICs of essential oil and nanoemulsion. Estragol was identified as the main component in the essential oil. The average diameter of nanoemulsion particle was 50 nm and the zeta potential was -30mV. The MIC values ​​ of essential oil and nanoemulsion were 0.58±0.11 and 0.33±0.07mg/ml, respectively, and their MBC were 0.65 ± 0.20 and 0.38 ± 0.15 mg/ml, respectively. Nanoamulsion had a greater inhibitory effect against bacterial growth than free essential oil. At the end of the 72-hour period, nanoemulsion treatment at 75% MIC resulted in a reduction in stx1A and stx2A transcription of 3.75 and 4.10 folds, while at 75% MIC of essential oil stx1A and stx2A transcripts were reduced 1.91 and 2.02 folds compared with control, respectively. Higher activity of nanoemulsion of tarragon essential oil to reduce the growth and shigatoxin production of E. coli compared to pure EO, reveals its potential to be used as a natural food preservative and a solution to the global problem of emergence of antibiotic-resistant microbes.

کلیدواژه‌ها English

Enterohemorrhagic E.coli
Essential oil
Nanoemulsion
Shigatoxin
Tarragon
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