Optimization of thymol nanoencapsulation in gelatin and carrageenan polymer network and investigation of anti-biofilm effect against marine bacteria Bacillus sp.

Document Type : Research Paper

Authors

1 Ph.D. in Marine Biology, Department of Marine Biology, Faculty of Marine Science and Technology, University of Hormozgan, Bandar Abbas, Iran

2 Professor in Department of Biology, Faculty of Science, University of Qom, Qom, Iran

3 Associate Professor in Department of Phytochemistry, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, Tehran, Iran

4 Associate Professor in Department of Marine Biology, Faculty of Marine Science and Technology, University of Hormozgan, Bandar Abbas, Iran

10.22124/japb.2023.22800.1477

Abstract

Thymol is the main monoterpene phenol occurring in essential oils. The aim of this study was to encapsulate thymol, in natural polymers gelatin and carrageenan. In order to achieve the optimization of encapsulated thymol, the CCD method of the experiment design software was used. As a result, based on the analysis, it was found that the optimal conditions suggested by Design Expert 13 software were as follows: pH 6, thymol concentration 0.4% (W/V) and the ratio of carrageenan to gelatin =1, which in this case, the encapsulation efficiency, nanocapsule size and polydispersity index were obtained 91%, 112 nm, and 1.1, respectively. The encapsulated thymol gave lower minimal inhibition concentration (MIC) and minimal bactericidal concentration (MBC) values than the unencapsulated thymol against Bacillus sp. strain. In addition, encapsulated thymol had 70% inhibition and eradication of biofilm in lower concentrations. In general, the results showed an improvement in antibacterial and antibiofilm activity for nanoencapsulated thymol against marine bacteria Bacillus sp. This formulation could be proposed as an alternative or adjuvant for controlling biofilm in marine bacteria in the first stage of the biofouling phenomenon.

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 Bannister J., Sievers M., Bush F. and Bloecher N. 2019. Biofouling in marine aquaculture: A review of recent research and developments. Biofouling, 35(6): 631–648. doi: 10.1080/08927014.2019.1640214
Braga P.C., Culici M., Alfieri M. and Dal Sasso M. 2008. Thymol inhibits Candida albicans biofilm formation and mature biofilm. International Journal of Antimicrobial Agents, 31: 472–477. doi: 10.1016/j.ijantimicag.2007.12.013
Da Silva Carvalho A.G., Da Costa Machado M.T., Barros H.D.D. F.Q., Cazarin C.B.B., Junior M.R.M. and Hubinger M.D. 2019. Anthocyanins from jussara (Euterpe edulis Martius) extract carried by calcium alginate beads pre-prepared using ionic gelation. Powder Technology, 345: 283–291. doi: 10.1016/j.powtec.2019.01.016
Dalleau S., Cateau E., Berges T., Berjeaud J.M. and Imbert C. 2008. In vitro activity of terpenes against Candida biofilms. International Journal of Antimicrobial Agents, 31: 572–576. doi: 10.1016/j.ijantimicag.2008.01.028
Di Pasqua R., Mamone G., Ferranti P., Ercolini D. and Mauriello G. 2010. Changes in the proteome of Salmonella enterica serovar Thompson as stress adaptation to sublethal concentrations of thymol. Proteomics, 10(5): 1040–1049. doi: 10.1002/pmic.200900568
Ghaderi L., Aliahmadi A., Ebrahimi S.N. and Rafati H. 2021. Effective inhibition and eradication of Pseudomonas aeruginosa biofilms by Satureja khuzistanica essential oil nanoemulsion. Journal of Drug Delivery Science and Technology, 61: 1–8 (102260). doi: 10.1016/j.jddst.2020.102260
Gilsenan P. and Ross-Murphy S. 2000. Rheological characterization of gelatins from mammalian                 and marine sources. Food Hydrocolloids, 14: 191–195. doi: 10.1016/S0268-005X(99)00050-8
Holdt S.L. and Kraan S. 2011. Bioactive compounds in seaweed: Functional food applications and legislation. Journal of Applied Phycology, 23: 543–597. doi: 10.1007/s10811-010-9632-5
Hsieh W.C., Chang C.P. and Gao Y.L. 2006. Controlled release properties of chitosan encapsulated volatile citronella oil microcapsules by thermal treatments. Colloids and Surfaces, 53: 209–214. doi: 10.1016/j.colsurfb.2006.09.008
Hu K., Huang X., Gao Y., Huang X., Xiao H. and McClements D.J. 2015. Core-shell biopolymer nanoparticle delivery systems: Synthesis and characterization of curcumin fortified zein-pectin nanoparticles. Food Chemistry, 182: 275–281. doi: 10.1016/j.foodchem.2015.03.009
Jafri H., Ansari F.A. and Ahmad I. 2019. Prospects of essential oils in controlling pathogenic biofilm. P: 203–236. In: Ahmad Khan M.S., Ahmad I. and Chattopadhyay D. (Eds.). New Look to Phytomedicine. Academic Press, USA. doi: 10.1016/B978-0-12-814619-4.00009-4
Kang J., Liu L., Wu X., Sun Y. and Liu Z. 2018. Effect of thyme essential oil against Bacillus cereus planktonic growth and biofilm formation. Applied Microbiology and Biotechnology, 102: 10209–10218. doi: 10.1007/s00253-018-9401-y
Ludensky M. 1998. An automated system for biocide testing on biofilms. Journal of Industrial Microbiology and Biotechnology, 20(2): 109–115. doi: 10.1038/sj.jim.2900487
Mak W., Hamid N., Liu T., Lu J. and White W. 2013. Fucoidan from New Zealand Undaria pinnatifida: Monthly variations and determination of antioxidant activities. Carbohydrate Polymers, 95(1): 606–614. doi: 10.1016/j.carbpol.2013.02.047
Patel A., Hu Y., Tiwari J.K. and Velikov K.P. 2010. Synthesis and characterization of zein–curcumin colloidal particles. Soft Matter, 6: 6192–6199. doi: 10.1039/C0SM00800A
Raei P., Pourlak T., Memar M.Y., Alizadeh N., Aghamali M., Zeinalzadeh E., Asgharzadeh M. and Kafil H. 2017. Thymol and carvacrol strongly inhibit biofilm formation and growth of carbapenemase-producing Gram negative bacilli. Cellular and Molecular Biology, 63(5): 108–112. doi: 10.14715/cmb/2017.63.5.20
Rajitha Z., Nancharaiah Y.V. and Venugopalan V.P. 2020. Insight into bacterial biofilm-barnacle larvae interactions for environmentally benign antifouling strategies. International Biodeterioration and Biodegradation, 149: 1–12 (104937). doi: 10.1016/j.ibiod.2020.104937
Rassu G., Nieddu M., Bosi P., Trevisi P., Colombo M., Priori D., Manconi P., Giunchedi P., Gavini E. and Boatto G. 2014. Encapsulation and modified-release of thymol from oral microparticles as adjuvant or substitute to current medications. Phytomedicine, 21(21): 1627–1632. doi: 10.1016/j.phymed.2014.07.017
Sun X., Pan C., Ying Z., Yu D., Duan X., Huang F., Ling J. and Ouyang X.K. 2020. Stabilization of zein nanoparticles with k-carrageenan and tween 80 for encapsulation of curcumin. International Journal of Biological Macromolecules, 146: 549–559. doi: 10.1016/j.ijbiomac.2020.01.053
Tian Z., Lei Z., Chen X. and Chen Y.  2020. Evaluation of laser cleaning for defouling of marine biofilm contamination on aluminum alloys. Applied Surface Science, 499: 1–12 (144060). doi: 10.1016/j.apsusc.2019.144060
Wattanasatcha A., Rengpipat S. and Wanichwecharungruang S.  2012. Thymol nanospheres as an effective anti-bacterial agent. International Journal of Pharmaceutics, 434: 360–365. doi: 10.1016/j.ijpharm.2012.06.017
Zarei Jeliani Z., Sohrabipour J., Soltani M., Rabiei R. and Yousefzadi M. 2021. Seasonal variations in growth and phytochemical compounds of cultivated red alga, Hypnea flagelliformis, in southern coastlines of Iran. Journal of Applied Phycology, 33: 2459–2470. doi: 10.1007/s10811-021-02429-9
ZoBell C.E.  1941. Studies on marine bacteria. I. The cultural requirements of heterotrophic aerobes. Journal of Marine Research, 4: 41–75.