Antimicrobial and anti-biofilm effects of carotenoid pigment extracted from Rhodotorula glutinis strain on food-borne bacteria
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Abstract
Background and Objectives: Carotenoid pigments are among the most important pigments and have many applications in various food, cosmetics, hygiene industries and biotechnology. These pigments are produced by plants and microorganisms including Rhodotorula spp. This research intended to study the antimicrobial and antibiofilm effects of the carotenoid pigment from Rhodotorula glutinis on food spoilage bacteria (Staphylococcus aureus and Salmonella Typhimurium).
Materials and Methods: The R. glutinis was isolated from milk samples of cows with mastitis and ITS sequence-based typing was performed on them. After extracting the pigment from R. glutinis, its purity was examined using thin-layer chromatography. Following that, the broth microdilution method was used to evaluate antimicrobial effects of the pigment and MtP assay and subsequently scanning electron microscopy were used to assess the antibiofilm effects. In addition, the sub-MIC effects of the pigment on expression of quorum-sensing (QS) genes in S. Typhimurium isolates (sdiA and luxS) and S. aureus isolates (hld) were studied. Finally, the degree of toxicity of the pigment was analyzed using the MTT assay.
Results: ITS sequence analysis of R. glutinis revealed that the recently separated isolates exhibited strong differences with the strains recorded in NCBI database in genetic structure. The pigment produced by R. glutinis had strong antimicrobial effects and its mean MIC against S. Typhimurium isolates (17.0 μl.ml-1) was higher than the mean MIC against the S. aureus isolates (4.1 μl.ml-1). Electron microscope images and real-time observations indicated that the sub-MIC values of the pigment suppressed biofilm formation by suppressing expression of QS genes. In addition, the mentioned pigment at high MIC concentrations did not have toxic effects on Vero cells.
Conclusion: This research suggests that R. glutinis pigment is effective in destroying the planktonic form of food spoilage bacteria and degrading food spoilage biofilm-forming bacteria. Moreover, considering the low toxicity level of R. glutinis pigment for eukaryotic cells, we can suggest its use as a natural antibacterial preservative in various food materials.
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8. Asfour HZ. Anti-quorum sensing natural compounds. J Microsc Ultrastruct 2018; 6: 1-10.
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11. Sen T, Barrow CJ, Deshmukh SK. Microbial pigments in the food industry-challenges and the way forward. Front Nutr 2019; 6: 7.
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15. Poorniammal R, Prabhu S, Dufossé L, Kannan J. Safety evaluation of fungal pigments for food applications. J Fungi (Basel) 2021; 7: 692.
16. Rapoport A, Guzhova I, Bernetti L, Buzzini P, Kieliszek M, Kot AM. Carotenoids and some other pigments from fungi and yeasts. Metabolites 2021; 11: 92.
17. Hermanns AS, Zhou X, Xu Q, Tadmor Y, Li L. Carotenoid pigment accumulation in horticultural plants. Hortic Plant J 2020; 6: 343-360.
18. Novoveská L, Ross ME, Stanley MS, Pradelles R, Wasiolek V, Sassi J-F. Microalgal carotenoids: A review of production, current markets, regulations, and future direction. Mar Drugs 2019; 17: 640.
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21. Gosalawit C, Imsoonthornruksa S, Udomsil N, Ketudat-Cairns M. Genome sequence of the Oleaginous yeast Rhodotorula paludigena strain CM33, a potential strain for biofuel production. Microbiol Resour Announc 2020; 9(19): e00286-20.
22. Allahkarami S, Sepahi AA, Hosseini H, Razavi MR. Isolation and identification of carotenoid-producing Rhodotorula sp. from Pinaceae forest ecosystems and optimization of in vitro carotenoid production. Biotechnol Rep (Amst) 2021; 32: e00687.
23. Forgacs E, Cserhati T. Thin-layer chromatography of natural pigments: new advances. J Liq Chromatogr Relat Technol 2002; 25: 1521-1541.
24. CLSI. Performance Standards for Antimicrobial Susceptibility Testing. 30th ed. CLSI supplement M100. Wayne, PA: Clinical and Laboratory Standards Institute; 2020.
25. Kord M, Ardebili A, Jamalan M, Jahanbakhsh R, Behnampour N, Ghaemi EA. Evaluation of biofilm formation and presence of ica genes in Staphylococcus epidermidis clinical isolates. Osong Public Health Res Perspect 2018; 9: 160-166.
26. Kolar SL, Ibarra JA, Rivera FE, Mootz JM, Davenport JE, Stevens SM, et al. Extracellular proteases are key mediators of Staphylococcus aureus virulence via the global modulation of virulence‐determinant stability. Microbiologyopen 2013; 2: 18-34.
27. Karavolos MH, Bulmer DM, Winzer K, Wilson M, Mastroeni P, Williams P, et al. LuxS affects flagellar phase variation independently of quorum sensing in Salmonella enterica serovar Typhimurium. J Bacteriol 2008; 190: 769-771.
28. Neogi SB, Islam MM, Islam SKS, Akhter AHMT, Sikder MMH, Yamasaki S, et al. Risk of multi-drug resistant Campylobacter spp. and residual antimicrobials at poultry farms and live bird markets in Bangladesh. BMC Infect Dis 2020; 20: 278.
29. Mantravadi PK, Kalesh KA, Dobson RC, Hudson AO, Parthasarathy A. The Quest for novel antimicrobial compounds: emerging trends in research, development, and technologies. Antibiotics (Basel) 2019; 8: 8.
30. Rostami H, Hamedi H, Yolmeh M. Some biological activities of pigments extracted from Micrococcus roseus (PTCC 1411) and Rhodotorula glutinis (PTCC 5257). Int J Immunopathol Pharmacol 2016; 29: 684-695.
31. Tegegne DT, Yalew ST, Emeru BA, Messele EY,Werid GM, Bora SK, et al. Study of prevalence, associated risk factors and causative bacteria of bovine mastitis in Ethiopia. Int J Vet Sci Technol 2020; 4: 001-6.
32. Manimala MRA, Murugesan R. In vitro antioxidant and antimicrobial activity of carotenoid pigment extracted from Sporobolomyces sp. isolated from natural source. J Appl Nat Sci 2014; 6: 649-653.
33. Yolmeh M, Khomeiri M. Effect of mutagenesis treatment on antimicrobial and antioxidant activities of pigments extracted from Rhodotorula glutinis. Biocatal Agric Biotechnol 2017; 10: 285-290.
34. McKeegan KS, Borges-Walmsley MI, Walmsley AR. Microbial and viral drug resistance mechanisms. Trends Microbiol 2002; 10(10 Suppl): S8-14.
35. Ramesh C, Vinithkumar NV, Kirubagaran R. Marine pigmented bacteria: A prospective source of antibacterial compounds. J Nat Sci Biol Med 2019; 10: 104-113.
36. Bin L, Wei L, Xiaohong C, Mei J, Mingsheng D. In vitro antibiofilm activity of the melanin from Auricularia auricula, an edible jelly mushroom. Ann Microbiol 2012; 62: 1523-1530.
37. Wang K-L, Dou Z-R, Gong G-F, Li H-F, Jiang B, Xu Y. Anti-larval and anti-algal natural products from marine microorganisms as sources of anti-biofilm agents. Mar Drugs 2022; 20: 90.
38. Zhou K, Li C, Chen D, Pan Y, Tao Y, Qu W, et al. A review on nanosystems as an effective approach against infections of Staphylococcus aureus. Int J Nanomedicine 2018; 13: 7333-7347.
39. Ejaz R, Malik S, Ahmad M, Ali H, Choudhry S. Anti-biofilm potential of menthol purified from Mentha piperita L.(Mint). Biol Clin Sci Res J 2020; 2020(1): e037.
40. Oladejo O, Imani J. Inhibitory Effect of CUSTOS, a Formulated Allium-based extract, on the growth of some selected plant pathogens. Int J Plant Biol 2022; 13: 44-54.
41. Riyaz AL, Natarajan B, Monisha M, Paari KA. Probiotic properties of Enterococcus faecium isolated from Gallus gallus domesticus and its anti–microbial, anti–biofilm and growth enhancing potential in Danio rerio. J Fish Environ 2021; 45: 23-34.
42. Goel N, Fatima SW, Kumar S, Sinha R, Khare SK. Antimicrobial resistance in biofilms: Exploring marine actinobacteria as a potential source of antibiotics and biofilm inhibitors. Biotechnol Rep (Amst) 2021; 30: e00613.
43. Zhu H, He C-C, Chu Q-H. Inhibition of quorum sensing in Chromobacterium violaceum by pigments extracted from Auricularia auricular. Lett Appl Microbiol 2011; 52: 269-274.
44. Lee J-E, Lee N-K, Paik H-D. Antimicrobial and anti-biofilm effects of probiotic Lactobacillus plantarum KU200656 isolated from kimchi. Food Sci Biotechnol 2020; 30: 97-106.
45. Jin H-J, Zhang X, Cao H, Niu Y-J, Li C, Liu H. Chemical composition, security and bioactivity of the red pigment from Penicillium purpurogenum Li-3. Chem Biodivers 2018; 15(12): e1800300.
2. Schirone M, Visciano P, Tofalo R, Suzzi G. Editorial: Foodborne pathogens: Hygiene and safety. Front Microbiol 2019; 10: 1974.
3. Schneider G, Schweitzer B, Steinbach A, Pertics BZ, Cox A, Kőrösi L. Antimicrobial efficacy and spectrum of Phosphorous-Fluorine co-doped TiO2 nanoparticles on the foodborne pathogenic bacteria Campylobacter jejuni, Salmonella Typhimurium, Enterohaemorrhagic E. coli, Yersinia enterocolitica, Shewanella putrefaciens, Listeria monocytogenes and Staphylococcus aureus. Foods 2021; 10: 1786.
4. Augustin J-C, Kooh P, Bayeux T, Guillier L, Meyer T, Jourdan-Da Silva N, et al. Contribution of foods and poor food-handling practices to the burden of foodborne infectious diseases in France. Foods 2020; 9: 1644.
5. Serwecińska L. Antimicrobials and antibiotic-resistant bacteria: a risk to the environment and to public health. Water 2020; 12: 3313.
6. Zhou L, Zhang Y, Ge Y, Zhu X, Pan J. Regulatory mechanisms and promising applications of quorum sensing-inhibiting agents in control of bacterial biofilm formation. Front Microbiol 2020; 11: 589640.
7. Jiang Q, Chen J, Yang C, Yin Y, Yao K. Quorum sensing: a prospective therapeutic target for bacterial diseases. Biomed Res Int 2019; 2019: 2015978.
8. Asfour HZ. Anti-quorum sensing natural compounds. J Microsc Ultrastruct 2018; 6: 1-10.
9. Hakimi Alni R, Ghorban K, Dadmanesh M. Combined effects of Allium sativum and Cuminum cyminum essential oils on planktonic and biofilm forms of Salmonella Typhimurium isolates. 3 Biotech 2020; 10: 315.
10. Choksi J, Vora J, Shrivastava N. Bioactive pigments from isolated bacteria and its antibacterial, antioxidant and sun protective application useful for cosmetic products. Indian J Microbiol 2020; 60: 379-382.
11. Sen T, Barrow CJ, Deshmukh SK. Microbial pigments in the food industry-challenges and the way forward. Front Nutr 2019; 6: 7.
12. Keller NP. Fungal secondary metabolism: regulation, function and drug discovery. Nat Rev Microbiol 2019; 17: 167-180.
13. Singh BP, Rateb ME, Rodriguez-Couto S, Polizeli MLTM, Li W-J. Editorial: Microbial secondary metabolites: recent developments and technological challenges. Front Microbiol 2019; 10: 914.
14. Yadav AN. Microbial biotechnology for bio-prospecting of microbial bioactive compounds and secondary metabolites. J Appl Biol Biotechnol 2021; 9: 1-6.
15. Poorniammal R, Prabhu S, Dufossé L, Kannan J. Safety evaluation of fungal pigments for food applications. J Fungi (Basel) 2021; 7: 692.
16. Rapoport A, Guzhova I, Bernetti L, Buzzini P, Kieliszek M, Kot AM. Carotenoids and some other pigments from fungi and yeasts. Metabolites 2021; 11: 92.
17. Hermanns AS, Zhou X, Xu Q, Tadmor Y, Li L. Carotenoid pigment accumulation in horticultural plants. Hortic Plant J 2020; 6: 343-360.
18. Novoveská L, Ross ME, Stanley MS, Pradelles R, Wasiolek V, Sassi J-F. Microalgal carotenoids: A review of production, current markets, regulations, and future direction. Mar Drugs 2019; 17: 640.
19. Sharifi A, Mohammadzadeh A, Salehi TZ, Mahmoodi P, Nourian A. Cuminum cyminum L. essential oil: A promising antibacterial and antivirulence agent against multidrug-resistant Staphylococcus aureus. Front Microbiol 2021; 12: 667833.
20. White TJ, Bruns TD, Lee SB, Taylor JW (1990). Amplification and Direct Sequencing of Fungal Ribosomal RNA Genes for Phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ, Eds. PCR Protocols: A Guide Methods Appli, Academic Press, New York, 315-322.
21. Gosalawit C, Imsoonthornruksa S, Udomsil N, Ketudat-Cairns M. Genome sequence of the Oleaginous yeast Rhodotorula paludigena strain CM33, a potential strain for biofuel production. Microbiol Resour Announc 2020; 9(19): e00286-20.
22. Allahkarami S, Sepahi AA, Hosseini H, Razavi MR. Isolation and identification of carotenoid-producing Rhodotorula sp. from Pinaceae forest ecosystems and optimization of in vitro carotenoid production. Biotechnol Rep (Amst) 2021; 32: e00687.
23. Forgacs E, Cserhati T. Thin-layer chromatography of natural pigments: new advances. J Liq Chromatogr Relat Technol 2002; 25: 1521-1541.
24. CLSI. Performance Standards for Antimicrobial Susceptibility Testing. 30th ed. CLSI supplement M100. Wayne, PA: Clinical and Laboratory Standards Institute; 2020.
25. Kord M, Ardebili A, Jamalan M, Jahanbakhsh R, Behnampour N, Ghaemi EA. Evaluation of biofilm formation and presence of ica genes in Staphylococcus epidermidis clinical isolates. Osong Public Health Res Perspect 2018; 9: 160-166.
26. Kolar SL, Ibarra JA, Rivera FE, Mootz JM, Davenport JE, Stevens SM, et al. Extracellular proteases are key mediators of Staphylococcus aureus virulence via the global modulation of virulence‐determinant stability. Microbiologyopen 2013; 2: 18-34.
27. Karavolos MH, Bulmer DM, Winzer K, Wilson M, Mastroeni P, Williams P, et al. LuxS affects flagellar phase variation independently of quorum sensing in Salmonella enterica serovar Typhimurium. J Bacteriol 2008; 190: 769-771.
28. Neogi SB, Islam MM, Islam SKS, Akhter AHMT, Sikder MMH, Yamasaki S, et al. Risk of multi-drug resistant Campylobacter spp. and residual antimicrobials at poultry farms and live bird markets in Bangladesh. BMC Infect Dis 2020; 20: 278.
29. Mantravadi PK, Kalesh KA, Dobson RC, Hudson AO, Parthasarathy A. The Quest for novel antimicrobial compounds: emerging trends in research, development, and technologies. Antibiotics (Basel) 2019; 8: 8.
30. Rostami H, Hamedi H, Yolmeh M. Some biological activities of pigments extracted from Micrococcus roseus (PTCC 1411) and Rhodotorula glutinis (PTCC 5257). Int J Immunopathol Pharmacol 2016; 29: 684-695.
31. Tegegne DT, Yalew ST, Emeru BA, Messele EY,Werid GM, Bora SK, et al. Study of prevalence, associated risk factors and causative bacteria of bovine mastitis in Ethiopia. Int J Vet Sci Technol 2020; 4: 001-6.
32. Manimala MRA, Murugesan R. In vitro antioxidant and antimicrobial activity of carotenoid pigment extracted from Sporobolomyces sp. isolated from natural source. J Appl Nat Sci 2014; 6: 649-653.
33. Yolmeh M, Khomeiri M. Effect of mutagenesis treatment on antimicrobial and antioxidant activities of pigments extracted from Rhodotorula glutinis. Biocatal Agric Biotechnol 2017; 10: 285-290.
34. McKeegan KS, Borges-Walmsley MI, Walmsley AR. Microbial and viral drug resistance mechanisms. Trends Microbiol 2002; 10(10 Suppl): S8-14.
35. Ramesh C, Vinithkumar NV, Kirubagaran R. Marine pigmented bacteria: A prospective source of antibacterial compounds. J Nat Sci Biol Med 2019; 10: 104-113.
36. Bin L, Wei L, Xiaohong C, Mei J, Mingsheng D. In vitro antibiofilm activity of the melanin from Auricularia auricula, an edible jelly mushroom. Ann Microbiol 2012; 62: 1523-1530.
37. Wang K-L, Dou Z-R, Gong G-F, Li H-F, Jiang B, Xu Y. Anti-larval and anti-algal natural products from marine microorganisms as sources of anti-biofilm agents. Mar Drugs 2022; 20: 90.
38. Zhou K, Li C, Chen D, Pan Y, Tao Y, Qu W, et al. A review on nanosystems as an effective approach against infections of Staphylococcus aureus. Int J Nanomedicine 2018; 13: 7333-7347.
39. Ejaz R, Malik S, Ahmad M, Ali H, Choudhry S. Anti-biofilm potential of menthol purified from Mentha piperita L.(Mint). Biol Clin Sci Res J 2020; 2020(1): e037.
40. Oladejo O, Imani J. Inhibitory Effect of CUSTOS, a Formulated Allium-based extract, on the growth of some selected plant pathogens. Int J Plant Biol 2022; 13: 44-54.
41. Riyaz AL, Natarajan B, Monisha M, Paari KA. Probiotic properties of Enterococcus faecium isolated from Gallus gallus domesticus and its anti–microbial, anti–biofilm and growth enhancing potential in Danio rerio. J Fish Environ 2021; 45: 23-34.
42. Goel N, Fatima SW, Kumar S, Sinha R, Khare SK. Antimicrobial resistance in biofilms: Exploring marine actinobacteria as a potential source of antibiotics and biofilm inhibitors. Biotechnol Rep (Amst) 2021; 30: e00613.
43. Zhu H, He C-C, Chu Q-H. Inhibition of quorum sensing in Chromobacterium violaceum by pigments extracted from Auricularia auricular. Lett Appl Microbiol 2011; 52: 269-274.
44. Lee J-E, Lee N-K, Paik H-D. Antimicrobial and anti-biofilm effects of probiotic Lactobacillus plantarum KU200656 isolated from kimchi. Food Sci Biotechnol 2020; 30: 97-106.
45. Jin H-J, Zhang X, Cao H, Niu Y-J, Li C, Liu H. Chemical composition, security and bioactivity of the red pigment from Penicillium purpurogenum Li-3. Chem Biodivers 2018; 15(12): e1800300.
| Files | ||
| Issue | Vol 15 No 1 (2023) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v15i1.11922 | |
| Keywords | ||
| Rhodotorula glutinis; Food-borne bacteria; Carotenoid pigment; Quorum sensing; Anti-biofilm | ||
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This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Naisi S, Bayat M, Zahraei SalehiT, Rahimian Zarif B, Yahyaraeyat R. Antimicrobial and anti-biofilm effects of carotenoid pigment extracted from Rhodotorula glutinis strain on food-borne bacteria. Iran J Microbiol. 2023;15(1):79-88.
