تاثیر آلکالوئید توماتیدین بر تشکیل بیوفیلم و بیان ژن های موثر در حساسیت جمعیتی سودوموناس آئروژینوزا
الموضوعات :
هادی قوم دوست نوری
1
,
امید تجربه کار
2
,
الهام معظمیان
3
1 - دانشجوی دکتری، گروه میکروبیولوژی، دانشکده علوم، کشاورزی و فناوری نوین، واحد شیراز، دانشگاه آزاد اسلامی، شیراز، ایران.
2 - استادیار، گروه میکروبیولوژی پزشکی (باکتریولوژی و ویروس شناسی)، دانشکده پزشکی افضلی پور، دانشگاه علوم پزشکی کرمان، کرمان، ایران.
3 - دانشیار، گروه میکروبیولوژی، دانشکده علوم، کشاورزی و فناوری نوین، واحد شیراز، دانشگاه آزاد اسلامی، شیراز، ایران.
الکلمات المفتاحية:
ملخص المقالة :
سابقه و هدف: سیستم حساسیت جمعیتی نقش مهمی در تنظیم فاکتورهای بیماری زایی و تشکیل بیوفیلم در سودوموناس آئروژینوزا بازی می کند. بنابراین مهار سیستم حساسیت جمعیتی می تواند راه حل مناسبی در درمان عفونت های مرتبط با بیوفیلم سودوموناس آئروژینوزا باشد. این مطالعه به منظور بررسی اثر غلظت های زیر حد مهاری توماتیدین بر بیان ژن های حساسیت جمعیتی و تشکیل بیوفیلم در سودوموناس آئروژینوزا انجام شد. مواد و روش ها: مطاله حاضر از نوع مداخله ای آزمایشگاهی است. ارزیابی تشکیل بیوفیلم با استفاده از روش میکروتیتر پلیت انجام شد.RNA تمام باکتری ها با استفاده از کیت ستونی جداسازی RNA، استخراج و بیان ژن های lasI، lasR، rhlI، rhlR وalgD در دو شرایط نرمال و تیمار با توماتیدین مورد بررسی قرار گرفت. ارزیابی مقاومت آنتی بیوتیکی به روش انتشار دیسک صورت گرفت. یافته ها: بیشترین و کمترین مقاومت آنتی بیوتیکی به ترتیب در برابر تیکارسیلین (%80) و کلیستین (%5) بود. توماتیدین تا غلظت 2 میلی گرم بر میلی لیتر فاقد اثر مهاری بر روی سودوموناس آئروژینوزا بود. 55% سویه ها تولید کننده بیوفیلم ضعیف و 45% تولیدکننده بیوفیلم قوی بودند. بیان همه ژن ها و تشکیل بیوفیلم پس از تیمار با توماتیدین افزایش یافت. ژن هایlasI وrhlR به ترتیب بیشترین و کمترین ژن های تحت تاثیر با توماتیدین بودند. در میان پنج ژن مورد بررسی، تنها rhlR در تولید کنندگان بیوفیلم ضعیف به طور قابل توجهی بیشتر بیان شد. نتیجه گیری: نتایج نشان داد غلظت های زیر حد مهاری توماتیدین، بر خلاف انتظار باعث افزایش بیان ژن های حساسیت جمعیتی و افزایش تشکیل بیوفیلم در سودوموناس آئروژینوزا می شود.
1-Tarkashvand, M., Lakzian, A., Fotovat, A., Mohammady, M. Isolation screening, and efficiency of Psudomonas isolates in biofilm formation on organic and inorganic carriers and phenanthrene bioremediation. Journal of Microbial World. 2020; 13(1): 6-20.
2-Ranieri, M.R., Whitchurch, C.B., Burrows, L.L. Mechanisms of biofilm stimulation by subinhibitory concentrations of antimicrobials. Curr Opin Microbiol. 2018; 45: 164-169.
3-Thi, M. T. T., Wibowo, D., & Rehm, B. H. Pseudomonas aeruginosa biofilms. International journal of molecular sciences. 2020; 21(22): 8671.
4-Firouzi Dalvand, L., Hosseini, F., Moradi Dehaghi, Sh., Siasi Torbati, E. Antibacterial and antibiofilm activity of bismuth oxide nanoparticles produced by bacillus subtilis against clinical Pseudomonas aeruginosa isolated from wound infections. Journal of Microbial World. 2019; 12(2): 172-185.
5-Saxena, P., Joshi, Y., Rawat, K., Bisht, R. Biofilms: architecture, resistance, quorum sensing and control mechanisms. Indian J. Microbiol. 2019; 59(1): 3-12.
6-Tavafi H, Abdi-Ali A, Ghadam P, Gharavi S. Evaluation of the synergistic effect of bacterial recombinant alginate lyase and therapeutic antibiotics on the growth of planktonic Pseudomonas aeruginosa. Journal of Microbial World. 2019 Aug 23;12(2):160-71.
7-Smith, R.S., Iglewski, B.H. P. aeruginosa quorum-sensing systems and virulence. Curr. Opin. Microbiol. 2003; 6(1): 56-60.
8-T. Oluwabusola, E., Katermeran, N P., Han Poh, W., Goh, T M B., Tan. L T., Diyaolu, O., Tabudravu, J., Ebel, R., A. Rice, S., Jaspars, M. Inhibition of the quorum sensing system, elastase production and biofilm formation in Pseudomonas aeruginosa by sammaplin A and Bisaprasin. Molecules. 2022; 27(1721): 1-17.
9-Schuster, M., Greenberg, E.P. A network of networks: quorum-sensing gene regulation in Pseudomonas aeruginosa. Int. J. Med. Microbiol.2018; 296(2-3): 73-81.
10-Rajkumari, J., Borkotoky, S., Murali, A., Suchiang, K., Mohanty, S.K., Busi, S. Cinnamic acid attenuates quorum sensing associated virulence factors and biofilm formation in Pseudomonas aeruginosa PAO1. Biotechnol Lett. 2018; 40(7): 1087-1100.
11-Almohaywi, B., Tin Yu, T., Iskander, G., Sabir, Sh., Bhadbhade, M., Blak, D., Kumar, N. Synthesis of alkyne-substituted dihydropyrrolones as bacterial quorum-sensing inhibitors of Pseudomonas aeruginosa. Antibiotics. 2022; 11(151): 1-16.
12-Lima, M.R., Ferreira, G.F., Neto, W.R.Nunes, Monteiro, J.M., Santos, A.R.C., Tavares, P. B., Denadai, A.M.L., Bomfim, M.R.Q., Santos, V.L.Dos, Marques, S.G., de Souza Monteiro, A. Evaluation of the interaction between polymyxin B and Pseudomonas aeruginosa biofilm and planktonic cells: reactive oxygen species induction and zeta potential. BMC Microbiol.2019; 19(1): 115.
13-Falahati, N., Jamali, H., Kargar, M. and Kafilzadeh, F., 2022. The evaluation of the effect of gold nanoparticles on the expression of (MexA/B) efflux pump genes from the RND family in multidrug resistant Pseudomonas aeruginosa strains. Journal of Microbial World, 15(1), pp.78-87.
14-Li, Q., Mao, S., Wang, H., Ye, X. The molecular architecture of Pseudomonas aeruginosa quorum-sensing inhibitors. Mar. Drugs. 2022; 20(488): 1-28.
15-Cushnie, T.P., Cushnie, B., Lamb, A.J. Alkaloids: an overview of their antibacterial, antibiotic-enhancing and antivirulence activities. Int. J. Antimicrob. Agents. 2014; 44(5): 377-386.
16-Lamontagne Boulet, M., Isabelle, C., Guay, I., Brouillette, E., Langlois, J.P., Jacques, P.E., Rodrigue, S., Brzezinski, R., Beauregard, P.B., Bouarab, K., Boyapelly, K., Boudreault, P.L., Marsault, E., Malouin, F. Tomatidine is a lead antibiotic molecule that targets Staphylococcus aureus ATP synthase subunit C. Antimicrob. Agents Chemother. 2018; 62(6).
17-Normandin, C., Boudreault, P.L. Concise large-scale synthesis of tomatidine, a potent antibiotic natural product. Molecules. 2021; 26(19): 6008.
18-Karami, P., Khaledi, A., Mashoof, R.Y., Yaghoobi, M.H., Karami, M., Dastan, D., Alikhani, M.Y. The correlation between biofilm formation capability and antibiotic resistance pattern in Pseudomonas aeruginosa. Gene Rep. 2020; 18: 100561.
19-Weinstein, M.P. Performance Standards for Antimicrobial Susceptibility Testing. Clinical and Laboratory Standards Institute. 2021
20-Wikler, M. Methods for dilution antimicrobial susceptibility test for bacteria that grow aerobically. In: Approved Standard 10th ed. M07-A11. 2018.
21-Kirmusaoglu, S. The methods for detection of biofilm and screening antibiofilm activity of agents. In: Antimicrobials, Antibiotic Resistance, Antibiofilm Strategies and Activity Methods, 15th ed. IntechOpen, pp. 2019; 1-17.
22-Mitchell, G., Lafrance, M., Boulanger, S., Seguin, D.L., Guay, I., Gattuso, M., Marsault, E., Bouarab, K., Malouin, F. Tomatidine acts in synergy with aminoglycoside antibiotics against multiresistant Staphylococcus aureus and prevents virulence gene expression. J. Antimicrob. Chemother. 2012 67(3): 559-568.
23-Hnamte, S., Parasuraman, P., Ranganathan, S., Ampasala, D.R., Reddy, D., Kumavath, R. N., Suchiang, K., Mohanty, S.K., Busi, S. Mosloflavone attenuates the quorum sensing controlled virulence phenotypes and biofilm formation in Pseudomonas aeruginosa PAO1: In vitro, in vivo and in silico approach. Microb. Pathog. 2019; 131: 128-134.
24-Schmittgen, T.D., Livak, K.J. Analyzing real-time PCR data by the comparative C (T) method. Nat. Protoc. 2008; 3(6): 1101-1108.
25-Ahmed, S., Rudden, M., Smyth, T.J., Dooley, J.S.G., Marchant, R., Banat, I.M. Natural quorum sensing inhibitors effectively downregulate gene expression of Pseudomonas aeruginosa virulence factors. Appl. Microbiol. Biotechnol. 2019; 103(8): 3521-3535.
26-Townsley, L., Shank, E.A. Natural-product antibiotics: cues for modulating bacterial biofilm formation. Trends Microbiol. 2017; 25 (12), 1016–1026.
27-Abd El-Baky, R.M., Masoud, S.M., Mohamed, D.S., Waly, N.G., Shafik, E.A., Mohareb, D. A., Elkady, A., Elbadr, M.M., Hetta, H.F. Prevalence and some possible mechanisms of colistin resistance among multidrug-resistant and extensively drug- resistant Pseudomonas aeruginosa. Infect. Drug Resist. 2020; 13: 323-332.
28-Abbas, H.A., El-Ganiny, A.M., Kamel, H.A. Phenotypic and genotypic detection of antibiotic resistance of Pseudomonas aeruginosa isolated from urinary tract infections. Afr. Health Sci. 2018: 18(1); 11-21.
29-Yamani, L., Alamri, A., Alsultan, A., Alfifi, S., Ansari, M.A., Alnimr, A. Inverse correlation between biofilm production efficiency and antimicrobial resistance in clinical isolates of Pseudomonas aeruginosa. Microb. Pathog. 2021; 157: 104989.
30-Morgan, S.J., Lippman, S.I., Bautista, G.E., Harrison, J.J., Harding, C.L., Gallagher, L.A., Cheng, A.C., Siehnel, R., Ravishankar, S., Usui, M.L., Olerud, J.E., Fleckman, P., Wolcott, R.D., Manoil, C., Singh, P.K., 2019. Bacterial fitness in chronic wounds appears to be mediated by the capacity for high-density growth, not virulence or biofilm functions. PLoS Pathog. 2019; 15(3): e1007511.
31-Sun, Z., Jiao, X., Peng, Q., Jiang, F., Huang, Y., Zhang, J., Yao, F. Antibiotic resistance in Pseudomonas aeruginosa is associated with decreased fitness. Cell Physiol. Biochem. 2013; 31(2-3): 347-354.
32-Pearson, J.P., Pesci, E.C., Iglewski, B.H. Roles of Pseudomonas aeruginosa las and rhl quorum-sensing systems in control of elastase and rhamnolipid biosynthesis genes. J. Bacteriol. 1997; 179(18): 5756-5767.
33-Kostylev, M., Kim, D.Y., Smalley, N.E., Salukhe, I., Greenberg, E.P., Dandekar, A.A. Evolution of the Pseudomonas aeruginosa quorum-sensing hierarchy. Proc. Natl. Acad. Sci. U. S. A. 2019; 116(14): 7027-7032.
34-Davies, J., Spiegelman, G.B., Yim, G. The world of subinhibitory antibiotic concentrations. Curr. Opin. Microbiol. 2006; 9(5): 445-453.
35-Guay, I., Boulanger, S., Isabelle, C., Brouillette, E., Chagnon, F., Bouarab, K., Marsault, E., Malouin, F., 2018. Tomatidine and analog FC04-100 possess bactericidal activities against Listeria, Bacillus and Staphylococcus spp. BMC Pharmacol Toxicol. 2018: 19(1): 7.
_||_1-Tarkashvand, M., Lakzian, A., Fotovat, A., Mohammady, M. Isolation screening, and efficiency of Psudomonas isolates in biofilm formation on organic and inorganic carriers and phenanthrene bioremediation. Journal of Microbial World. 2020; 13(1): 6-20.
2-Ranieri, M.R., Whitchurch, C.B., Burrows, L.L. Mechanisms of biofilm stimulation by subinhibitory concentrations of antimicrobials. Curr Opin Microbiol. 2018; 45: 164-169.
3-Thi, M. T. T., Wibowo, D., & Rehm, B. H. Pseudomonas aeruginosa biofilms. International journal of molecular sciences. 2020; 21(22): 8671.
4-Firouzi Dalvand, L., Hosseini, F., Moradi Dehaghi, Sh., Siasi Torbati, E. Antibacterial and antibiofilm activity of bismuth oxide nanoparticles produced by bacillus subtilis against clinical Pseudomonas aeruginosa isolated from wound infections. Journal of Microbial World. 2019; 12(2): 172-185.
5-Saxena, P., Joshi, Y., Rawat, K., Bisht, R. Biofilms: architecture, resistance, quorum sensing and control mechanisms. Indian J. Microbiol. 2019; 59(1): 3-12.
6-Tavafi H, Abdi-Ali A, Ghadam P, Gharavi S. Evaluation of the synergistic effect of bacterial recombinant alginate lyase and therapeutic antibiotics on the growth of planktonic Pseudomonas aeruginosa. Journal of Microbial World. 2019 Aug 23;12(2):160-71.
7-Smith, R.S., Iglewski, B.H. P. aeruginosa quorum-sensing systems and virulence. Curr. Opin. Microbiol. 2003; 6(1): 56-60.
8-T. Oluwabusola, E., Katermeran, N P., Han Poh, W., Goh, T M B., Tan. L T., Diyaolu, O., Tabudravu, J., Ebel, R., A. Rice, S., Jaspars, M. Inhibition of the quorum sensing system, elastase production and biofilm formation in Pseudomonas aeruginosa by sammaplin A and Bisaprasin. Molecules. 2022; 27(1721): 1-17.
9-Schuster, M., Greenberg, E.P. A network of networks: quorum-sensing gene regulation in Pseudomonas aeruginosa. Int. J. Med. Microbiol.2018; 296(2-3): 73-81.
10-Rajkumari, J., Borkotoky, S., Murali, A., Suchiang, K., Mohanty, S.K., Busi, S. Cinnamic acid attenuates quorum sensing associated virulence factors and biofilm formation in Pseudomonas aeruginosa PAO1. Biotechnol Lett. 2018; 40(7): 1087-1100.
11-Almohaywi, B., Tin Yu, T., Iskander, G., Sabir, Sh., Bhadbhade, M., Blak, D., Kumar, N. Synthesis of alkyne-substituted dihydropyrrolones as bacterial quorum-sensing inhibitors of Pseudomonas aeruginosa. Antibiotics. 2022; 11(151): 1-16.
12-Lima, M.R., Ferreira, G.F., Neto, W.R.Nunes, Monteiro, J.M., Santos, A.R.C., Tavares, P. B., Denadai, A.M.L., Bomfim, M.R.Q., Santos, V.L.Dos, Marques, S.G., de Souza Monteiro, A. Evaluation of the interaction between polymyxin B and Pseudomonas aeruginosa biofilm and planktonic cells: reactive oxygen species induction and zeta potential. BMC Microbiol.2019; 19(1): 115.
13-Falahati, N., Jamali, H., Kargar, M. and Kafilzadeh, F., 2022. The evaluation of the effect of gold nanoparticles on the expression of (MexA/B) efflux pump genes from the RND family in multidrug resistant Pseudomonas aeruginosa strains. Journal of Microbial World, 15(1), pp.78-87.
14-Li, Q., Mao, S., Wang, H., Ye, X. The molecular architecture of Pseudomonas aeruginosa quorum-sensing inhibitors. Mar. Drugs. 2022; 20(488): 1-28.
15-Cushnie, T.P., Cushnie, B., Lamb, A.J. Alkaloids: an overview of their antibacterial, antibiotic-enhancing and antivirulence activities. Int. J. Antimicrob. Agents. 2014; 44(5): 377-386.
16-Lamontagne Boulet, M., Isabelle, C., Guay, I., Brouillette, E., Langlois, J.P., Jacques, P.E., Rodrigue, S., Brzezinski, R., Beauregard, P.B., Bouarab, K., Boyapelly, K., Boudreault, P.L., Marsault, E., Malouin, F. Tomatidine is a lead antibiotic molecule that targets Staphylococcus aureus ATP synthase subunit C. Antimicrob. Agents Chemother. 2018; 62(6).
17-Normandin, C., Boudreault, P.L. Concise large-scale synthesis of tomatidine, a potent antibiotic natural product. Molecules. 2021; 26(19): 6008.
18-Karami, P., Khaledi, A., Mashoof, R.Y., Yaghoobi, M.H., Karami, M., Dastan, D., Alikhani, M.Y. The correlation between biofilm formation capability and antibiotic resistance pattern in Pseudomonas aeruginosa. Gene Rep. 2020; 18: 100561.
19-Weinstein, M.P. Performance Standards for Antimicrobial Susceptibility Testing. Clinical and Laboratory Standards Institute. 2021
20-Wikler, M. Methods for dilution antimicrobial susceptibility test for bacteria that grow aerobically. In: Approved Standard 10th ed. M07-A11. 2018.
21-Kirmusaoglu, S. The methods for detection of biofilm and screening antibiofilm activity of agents. In: Antimicrobials, Antibiotic Resistance, Antibiofilm Strategies and Activity Methods, 15th ed. IntechOpen, pp. 2019; 1-17.
22-Mitchell, G., Lafrance, M., Boulanger, S., Seguin, D.L., Guay, I., Gattuso, M., Marsault, E., Bouarab, K., Malouin, F. Tomatidine acts in synergy with aminoglycoside antibiotics against multiresistant Staphylococcus aureus and prevents virulence gene expression. J. Antimicrob. Chemother. 2012 67(3): 559-568.
23-Hnamte, S., Parasuraman, P., Ranganathan, S., Ampasala, D.R., Reddy, D., Kumavath, R. N., Suchiang, K., Mohanty, S.K., Busi, S. Mosloflavone attenuates the quorum sensing controlled virulence phenotypes and biofilm formation in Pseudomonas aeruginosa PAO1: In vitro, in vivo and in silico approach. Microb. Pathog. 2019; 131: 128-134.
24-Schmittgen, T.D., Livak, K.J. Analyzing real-time PCR data by the comparative C (T) method. Nat. Protoc. 2008; 3(6): 1101-1108.
25-Ahmed, S., Rudden, M., Smyth, T.J., Dooley, J.S.G., Marchant, R., Banat, I.M. Natural quorum sensing inhibitors effectively downregulate gene expression of Pseudomonas aeruginosa virulence factors. Appl. Microbiol. Biotechnol. 2019; 103(8): 3521-3535.
26-Townsley, L., Shank, E.A. Natural-product antibiotics: cues for modulating bacterial biofilm formation. Trends Microbiol. 2017; 25 (12), 1016–1026.
27-Abd El-Baky, R.M., Masoud, S.M., Mohamed, D.S., Waly, N.G., Shafik, E.A., Mohareb, D. A., Elkady, A., Elbadr, M.M., Hetta, H.F. Prevalence and some possible mechanisms of colistin resistance among multidrug-resistant and extensively drug- resistant Pseudomonas aeruginosa. Infect. Drug Resist. 2020; 13: 323-332.
28-Abbas, H.A., El-Ganiny, A.M., Kamel, H.A. Phenotypic and genotypic detection of antibiotic resistance of Pseudomonas aeruginosa isolated from urinary tract infections. Afr. Health Sci. 2018: 18(1); 11-21.
29-Yamani, L., Alamri, A., Alsultan, A., Alfifi, S., Ansari, M.A., Alnimr, A. Inverse correlation between biofilm production efficiency and antimicrobial resistance in clinical isolates of Pseudomonas aeruginosa. Microb. Pathog. 2021; 157: 104989.
30-Morgan, S.J., Lippman, S.I., Bautista, G.E., Harrison, J.J., Harding, C.L., Gallagher, L.A., Cheng, A.C., Siehnel, R., Ravishankar, S., Usui, M.L., Olerud, J.E., Fleckman, P., Wolcott, R.D., Manoil, C., Singh, P.K., 2019. Bacterial fitness in chronic wounds appears to be mediated by the capacity for high-density growth, not virulence or biofilm functions. PLoS Pathog. 2019; 15(3): e1007511.
31-Sun, Z., Jiao, X., Peng, Q., Jiang, F., Huang, Y., Zhang, J., Yao, F. Antibiotic resistance in Pseudomonas aeruginosa is associated with decreased fitness. Cell Physiol. Biochem. 2013; 31(2-3): 347-354.
32-Pearson, J.P., Pesci, E.C., Iglewski, B.H. Roles of Pseudomonas aeruginosa las and rhl quorum-sensing systems in control of elastase and rhamnolipid biosynthesis genes. J. Bacteriol. 1997; 179(18): 5756-5767.
33-Kostylev, M., Kim, D.Y., Smalley, N.E., Salukhe, I., Greenberg, E.P., Dandekar, A.A. Evolution of the Pseudomonas aeruginosa quorum-sensing hierarchy. Proc. Natl. Acad. Sci. U. S. A. 2019; 116(14): 7027-7032.
34-Davies, J., Spiegelman, G.B., Yim, G. The world of subinhibitory antibiotic concentrations. Curr. Opin. Microbiol. 2006; 9(5): 445-453.
35-Guay, I., Boulanger, S., Isabelle, C., Brouillette, E., Chagnon, F., Bouarab, K., Marsault, E., Malouin, F., 2018. Tomatidine and analog FC04-100 possess bactericidal activities against Listeria, Bacillus and Staphylococcus spp. BMC Pharmacol Toxicol. 2018: 19(1): 7.
