اثر ضد بیوفیلمی عصاره نعناع بر روی باکتری سودوموناس آئروژینوزا جدا شده از بیماران بستری شهرستان تبریز

نویسندگان
پریا وقری
منصور آل طه
گروه زیست شناسی، دانشکده تحصیلات تکمیلی، واحد بناب، دانشگاه آزاد اسلامی، بناب، ایران
چکیده
مقدمه

بیوفیلم­‌ها ممکن است بر روی سطوح زنده یا غیر زنده تشکیل شوند و در محیط طبیعی، صنعتی و بیمارستان شایع باشند. سودوموناس آئروژینوزا عامل پاتوژنیک بوده و پتانسیل بالایی برای تشکیل بیوفیلم دارد. تحقیق در مورد داروهای گیاهی برای مقابله با بیوفیلم از اهمیت خاصی برخوردار است. هدف از این تحقیق تعیین اثر عصاره نعناع بر روی باکتری و بیوفیلم جدایه‌های سودوموناس آئروژینوزا و تعیین فراوانی ژن pelF و ارتباط آن با تشکیل بیوفیلم می­‌باشد.

مواد و روش­‌ها

208 سویه بالینی جمع­‌آوری شد. بعد از عصاره­‌گیری تاثیر آن بر روی رشد سودوموناس آئروژینوزا بررسی گردید و مقدار MIC و MBC عصاره نعناع تعیین شد. زمان مناسب و شدت تشکیل بیوفیلم به صورت ضعیف، متوسط و قوی سنجیده شد و کمترین غلظت مهار بیوفیلم توسط عصاره نعناع (BIC) تعیین گردید. سپس حضور ژن pelF توسط PCR بررسی شد.

یافته‌­ها

عصاره نعناع باعث تشکیل هاله عدم رشد بر روی باکتری سودوموناس آئروژینوزا گردید و غلظت MIC و MBC عصاره نعناع به ترتیب 50 و 100 میلی گرم بر میلی لیتر تعیین شد. زمان مناسب تشکیل بیوفیلم 48 ساعت تعیین گردید و کمترین غلظت مهار بیوفیلم توسط عصاره نعناع 50 میلی گرم بر میلی لیتر به دست آمد. صد درصد نمونه­‌های جدا شده دارای ژن PelF بودند و این ژن رابطه معنی­‌داری با تشکیل بیوفیلم نشان داد (0/0001 =P).

نتیجه‌گیری

احتمالا می­توان از عصاره نعناع در ترکیبات مختلف برای مهار باکتری سودوموناس آئروژینوزا و مهار تشکیل بیوفیلم آن استفاده کرد.
کلیدواژه‌ها
سودوموناس آئروژینوزا
بیوفیلم
عصاره نعناع
ژن Pel
PCR

عنوان مقاله English

Anti-biofilm effect of peppermint extract on Pseudomonas aeruginosa that isolated of hospitalized patients in Tabriz city

نویسندگان English

paria vaghri
Mansoor Aletaha
Department of Biology, Faculty of Graduate Studies, Bonab Branch, Islamic Azad University, Bonab, Iran
چکیده English

Introduction

Green biosynthesis of the metallic nanoparticles is considered as one of the effective method for cancer treatment with minimum side effects. The objective of this study was the evaluation of cytotoxicity of zinc oxide (ZnO) nanoparticles synthesized with Ceratonia siliqua extract on the breast cancer cells and assessment of anti-angiogenesis properties of these nanoparticles in the chorioallantoic model (CAM).

Methods & Materials

In this investigation, the MDA-MB231 cancer cells line was treated for 24, 48 and 72 hours at different concentrations of ZnO nanoparticles (31.2 to 250 µg/ml) synthesized with Ceratonia siliqua extract and the percentage of cell viability was evaluated using an MTT assay. In addition, to determine the anti-angiogenesis effect, Ross Fertilized Chicken eggs were used in the CAM assay. After 12 days, the vessel number, vessels length, embryo length, and weight were measured.

Results

The findings showed that the green synthesized ZnO nanoparticle induces a cytotoxicity effect on breast cancer cell line (P≤ 0.001) depended in dose and time. The IC50 values were found about 62.5 μg/ml.

Evaluation of angiogenesis in the CAM model showed that ZnO nanoparticles in the dose ratio used, significantly reduced the number of vessels (P≤0.01); (P≤0.001) and the length of the vessels (P≤0.001); (P≤0.05) and embryo size (P≤0.05) and embryo weight ((P≤0.05) only in the high-dose).

Conclusion

ZnO nanoparticles synthesized with Ceratonia siliqua extract having a cytotoxic effect on breast cancer cells and anti-angiogenic effect can be an appropriate candidate in cancer treatment.

کلیدواژه‌ها English

Pseudomonas aeruginosa
Biofilm
Peppermint extract
Pel gene
PCR
1. Hamilton AC. Medicinal plants, conservation and livelihoods. Biodivers Conserv. 2004;13(8):1477-517.
2. Giaouris E, Heir E, Hébraud M, Chorianopoulos N, Langsrud S, Møretrø T, et al. Attachment and biofilm formation by foodborne bacteria in meat processing environments: causes, implications, role of bacterial interactions and control by alternative novel methods. Meat Science. 2014;97(3):298-309.
3. Sandasi M, Leonard CM, Van Vuuren SF, Viljoen AM. Peppermint (Mentha piperita) inhibits microbial biofilms in vitro. S. Afr. J. Bot. 2011;77(1):80-5.
4. Valeriano C, de Oliveira TLC, de Carvalho SM, Cardoso MdG, Alves E, Piccoli RH. The sanitizing action of essential oil-based solutions against Salmonella enterica serotype Enteritidis S64 biofilm formation on AISI 304 stainless steel. Food Control. 2012;25(2):673-7.
5. Costerton JW, Cheng K, Geesey GG, Ladd TI, Nickel JC, Dasgupta M, et al. Bacterial biofilms in nature and disease. Annu Rev Microbiol. 1987;41(1):435-64.
6. Adabi M, Talebi Taher M, Arbabi L, Afshar M, Fathizadeh S, Minaeian S, et al. Determination of antibiotic resistance pattern of Pseudomonas aeruginosa strains isolated from patients with burn wounds. JAUMS. 2015;15(1):66-74.
7. Arora DS, Kaur J. Antimicrobial activity of spices. Int J Antimicrob Agents. 1999;12(3):257-62.
8. Zhou S, Koh H-L, Gao Y, Gong Z-y, Lee EJD. Herbal bioactivation: The good, the bad and the ugly. Life Sciences. 2004;74(8):935-68.
9. Kok CR, Yang J, Ciftci O, Hutkins R. Antibacterial effects of peppermint, Mentha piperita essential oil in free-form and in nanoparticles on Pseudomonas fluorescens. 2016.
10. Ahmed A. Antibacterial activity of Mentha piperita and Allium sativum against some of gram-ve bacteria. Al-Mustansiriyah J Sci. 2012;23(5):39-48.
11. McKay DL, Blumberg JB. A review of the bioactivity and potential health benefits of peppermint tea (Mentha piperita L.). Phytother Res. 2006;20(8):619-33.
12. Friedman M, Henika PR, Mandrell RE. Bactericidal activities of plant essential oils and some of their isolated constituents against Campylobacter jejuni, Escherichia coli, Listeria monocytogenes, and Salmonella enterica. J Food Prot. 2002;65(10):1545-60.
13. Friedman L, Kolter R. Genes involved in matrix formation in Pseudomonas aeruginosa PA14 biofilms. Mol Microbiol. 2004;51(3):675-90.
14. Vasseur P, Vallet-Gely I, Soscia C, Genin S, Filloux A. The pel genes of the Pseudomonas aeruginosa PAK strain are involved at early and late stages of biofilm formation. Microbiology. 2005;151(3):985-97.
15. Colvin KM, Irie Y, Tart CS, Urbano R, Whitney JC, Ryder C, et al. The Pel and Psl polysaccharides provide Pseudomonas aeruginosa structural redundancy within the biofilm matrix. Environ Microbiol. 2012;14(8):1913-28.
16. Sakuragi Y, Kolter R. Quorum-sensing regulation of the biofilm matrix genes (pel) of Pseudomonas aeruginosa. J Bacteriol. 2007;189(14):5383-6.
17. Baltch AL, Smith RP. Pseudomonas aeruginosa: infections and treatment. Pseudomonas aeruginosa: infections and treatment. 1994(12).
18. Chanawong A, M'zali FH, Heritage J, Lulitanond A, Hawkey PM. SHV-12, SHV-5, SHV-2a and VEB-1 extended-spectrum β-lactamases in Gram-negative bacteria isolated in a university hospital in Thailand. J Antimicrob Chemother. 2001;48(6):839-52.
19. Chaudhari V, Gunjal S, Mehta M. Antibiotic resistance patterns of Pseudomonas aeruginosa in a tertiary care hospital in Central India. Int J Med Sci Public Health. 2013;2(2):386-9.
20. Eriksen H, Iversen B, Aavitsland P. Prevalence of nosocomial infections in hospitals in Norway, 2002 and 2003. J Hosp Infect. 2005;60(1):40-5.
21. Johnson JM, Church GM. Alignment and structure prediction of divergent protein families: periplasmic and outer membrane proteins of bacterial efflux pumps. J Mol Biol Mol. 1999;287(3):695-715.
22. Schümann J, Bluethmann H, Tiegs G. Synergism of Pseudomonas aeruginosa exotoxin A with endotoxin, superantigen, or TNF results in TNFR1-and TNFR2-dependent liver toxicity in mice. Immunol Lett. 2000;74(2):165-72.
23. Valle DL, Andrade JI, Puzon JJM, Cabrera EC, Rivera WL. Antibacterial activities of ethanol extracts of Philippine medicinal plants against multidrug-resistant bacteria. APJTB. 2015;5(7):532-40.
24. Nyenje ME, Green E, Ndip RN. Biofilm formation and adherence characteristics of Listeria ivanovii strains isolated from ready-to-eat foods in Alice, South Africa. ScientificWorldJournal. 2012;2012.
25. Moskowitz SM, Foster JM, Emerson J, Burns JL. Clinically feasible biofilm susceptibility assay for isolates of Pseudomonas aeruginosa from patients with cystic fibrosis. J Clin Microbiol. 2004;42(5):1915-22.
26. Jabalameli F, Mirsalehian A, Khoramian B, Aligholi M, Khoramrooz SS, Asadollahi P, et al. Evaluation of biofilm production and characterization of genes encoding type III secretion system among Pseudomonas aeruginosa isolated from burn patients. Burns. 2012;38(8):1192-7.
27. Van Delden C, Iglewski BH. Cell-to-cell signaling and Pseudomonas aeruginosa infections. Emerg Infect Dis. 1998;4(4):551.
28. Farzaei L, Azizi A, Arji I, Rostami R. Bioactive Potential of Peppermint Extract Against Some Human Bacterial and Fungal Pathogens. Avicenna J Clin Microbiol Infect. 2015;2(4).
29. Pulipati S, Koushik OS, Babu PS. Phytochemical Analysis and Antibacterial Efficacy of Mentha piperita (L) Ethanolic Leaf Extract against Clinical Isolates of Uropathogens. Br Microbiol Res J. 2016;13(6).
30. Niculae M, Spînu M, Şandru CD, Brudaşcă F, Cadar D, SZAKACS B, et al. Antimicrobial potential of some Lamiaceae essential oils against animal multiresistant bacteria. Lucrări Scintifice Medicină Veterinară Timoara. 2009;42:170-5.
31. Ceylan O, Ugur A, Sarac N, Sahin MD. The antimicrobial and antibiofilm activities of Mentha x piperita L. essential oil. J Biosci Biotechnol Discov. 2014.
32. Lang M, Rodrigues S, Boulho R, Duteil E, Bazire A, Bedoux G. An Essential Oil Blend Prevents Pseudomonas aeruginosa PA01 from Forming Biofilms. J Bacteriol Parasitol. 2016;7(2):1000268.