اثرات آنتی‌باکتریال و آنتی‌بیوفیلم پروبیوتیک‌ها علیه باکتری Pseudomonas aeruginosa مقاوم به چند دارو از جدایه‌های بالینی

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه زیست‌شناسی، دانشکده علوم، دانشگاه دیالی، دیالی، 32001، عراق.

2 گروه زیست‌شناسی، دانشکده آموزش علوم خالص، دانشگاه دیالی، دیالی، 32001، عراق.

3 گروه زیست‌شناسی، دانشکده علوم، دانشگاه دیالی، دیالی، 32001، عراق

چکیده

هدف: مقاومت آنتی‌بیوتیکی یک مشکل عمده سلامت جهانی محسوب می‌شود که درمان مؤثر بسیاری از عفونت‌های باکتریایی، به‌ویژه عفونت‌های ناشی از Pseudomonas aeruginosa مقاوم به چند دارو (MDR)، را با چالش مواجه کرده است. از این‌رو، نیاز مبرمی به توسعه راهکارهای نوین برای مقابله با میکروارگانیسم‌های مقاوم به آنتی‌بیوتیک وجود دارد. پروبیوتیک‌ها مزایای بالقوه‌ای را در درمان اختلالات مختلف نشان داده‌اند. هدف این مطالعه بررسی اثرات آنتی‌باکتریال و آنتی‌بیوفیلم پروبیوتیک‌ها علیه جدایه‌های بالینی P. aeruginosa مقاوم به چند دارو بود.
مواد و روش‌ها: در این مطالعه، 20 جدایه P. aeruginosa مقاوم به چند دارو از مجموع 165 نمونه بالینی شامل نمونه‌های گوش، زخم، سوختگی و ادرار جداسازی شد. آزمون حساسیت آنتی‌بیوتیکی با استفاده از روش انتشار دیسک انجام گرفت. گونه‌های Lactobacillus از 15 نمونه طبیعی مختلف (7 نمونه شیر، 5 نمونه ماست و 3 نمونه واژینال) جدا شده و با استفاده از آزمون‌های بیوشیمیایی شناسایی و به‌وسیله سیستم Vitek 2 تأیید شدند. سوپرناتانت بدون سلول (CFS) برای ارزیابی فعالیت‌های آنتی‌باکتریال و آنتی‌بیوفیلم علیه جدایه‌های P. aeruginosa مقاوم به چند دارو، با استفاده از روش انتشار در چاهک آگار و آزمون میکروپلیت برای سنجش فعالیت ضد بیوفیلم به‌کار گرفته شد.
نتایج: از میان 15 نمونه طبیعی، 10 نمونه (3 شیر، 5 ماست و 2 واژینال) از نظر حضور گونه‌های Lactobacillus مثبت بودند. از میان 20 جدایه P. aeruginosa، تنها 7 جدایه تولیدکننده قوی بیوفیلم بودند. نتایج نشان داد که قوی‌ترین اثرات مهاری بر رشد سه جدایه P. aeruginosa (P7، P13 و P15) در غلظت حداقل مهارکنندگی (MIC) و زیر MIC (به‌ترتیب 25 و 5/12 میکروگرم بر میلی‌لیتر) مشاهده شد، در حالی‌که چهار جدایه دیگر (P3، P4، P11 وP18 ) و MIC و زیر MIC برابر با 50 و 25 میکروگرم بر میلی‌لیتر را نشان دادند. اثر مهاری CFS تنها علیه 7 جدایه P. aeruginosa مقاوم به چند دارو با بیشترین میزان تشکیل بیوفیلم بررسی شد و نتایج، اثر آنتی‌باکتریال قابل توجهی را نشان داد. در جدایه‌های حساس، قطر هاله عدم رشد بین 3 تا 12 میلی‌متر متغیر بود.
نتیجه‌گیری: از میان 10 جدایه Lactobacillus spp.، دو جدایه (با منشأ واژینال) هیچ‌گونه فعالیت مهاری علیه یک یا چند سویه P. aeruginosa مقاوم به چند دارو نشان ندادند. گونه‌های Lactobacillus جداشده از شیر و ماست دارای اثر مهاری بر رشد P. aeruginosa بودند. یافته‌ها در نهایت این مفهوم را تأیید می‌کنند که می‌توان از پروبیوتیک‌ها به‌عنوان جایگزینی برای آنتی‌بیوتیک‌ها در درمان باکتری‌های بیماری‌زای مقاوم به آنتی‌بیوتیک استفاده کرد.

کلیدواژه‌ها

عنوان مقاله [English]

Antibacterial and Antibiofilm effects of probiotic against Multidrug Resistant Pseudomonas aeruginosa bacteria from clinical isolates

نویسندگان [English]

  • Izdehar Mohammed Jasim 1
  • Ghassan Mohammed Jasim 2
  • Abbas Yaseen Hasan 3
1 Department of Biology, College of sciences, University of Diyala, Diyala, 32001, Iraq.
2 Department of Biology, College of education for pure sciences, University of Diyala, Diyala, 32001, Iraq
3 Department of Biology, College of sciences, University of Diyala, Diyala, 32001, Iraq
چکیده [English]

Objective
Resistance to antibiotics represents a significant worldwide health issue that exposes the effective treatment of several bacterial illnesses, particularly those induced by multi-drug-resistant (MDR) Pseudomonas aeruginosa. Consequently, there's an imperative necessity to develop novel techniques to address resistant to antibiotics microorganisms. Probiotics which have demonstrated potential advantages in several disorders. Objective: The aim of study is to investigating the antibacterial and antibiofilm effects of probiotic against MDR P. aeruginosa clinical isolates.

Materials and methods
Twenty MDR P. aeruginosa isolates isolated from 165 samples obtained from different clinical specimen (ear, wound, burn, and urine). Antibiotics susceptibility test was performed using disc diffusion method. Lactobacilli species were isolated from 15 different natural samples (7 milk, 5 curd and 3 vagina), and identified by biochemical tests and confirmed using vitek 2 system. Cell free supernatant (CFS) had been utilized for evaluating both antibiofilm and antibacterials efficacy against multidrug-resistant P. aeruginosa isolates through the agar well diffusion method and microtiter plate assay to assess antibiofilm activity.
Results
Out of 15 different natural samples, 10 (3 milk, 5 curd and 2 vagina) samples were positive for Lactobacilli spp. Among 20 isolates of P. aeruginosa only seven isolates were strong biofilm producer. The findings indicated that the most potent inhibitory effects on the development for three isolates of P. aeruginosa (P7, P13, and P15) were observed at minimum inhibitory concentration (MIC) and sub-MIC levels (25 and 12.5 µg/mL, respectively, and four isolates (P3, P4, P11, and P18) exhibited MIC and sub-MIC (50 and 25 µg/mL), respectively. The inhibitory effect of CFS were done against only 7 MDR P. aeruginosa isolates which highest biofilm formation, The results showed antibacterial effect. In the susceptible isolates, the zone of inhibition varied from 3 mm to 12 mm in diameter.

Conclusions
Among 10 Lactobacillus spp. isolates, two isolates (from vaginal sources) exhibited no inhibitory activity against one or more of the multidrug-resistant P. aeruginosa strains. Lactobacilli spp. Isolated from (milk and curd) that have inhibitory effect on growth against P. aeruginosa, Findings ultimately corroborated the concept of employing probiotics as a substitute for antibiotics in the treatment of resistant to antibiotics pathogenic bacteria.

کلیدواژه‌ها [English]

  • biofilm
  • inhibitory effect
  • MDR
  • probiotics
  • Pseudomonas aeruginosa

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مجله بیوتکنولوژی کشاورزی
دوره 18، شماره 1
فروردین 1405
صفحه 399-418

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