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Antibacterial Activity of Sustainable Thymol Nanoemulsion Formulations Against the Bacterial Blight Disease on Cluster Bean Caused by Xanthomonas axonopodis

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Abstract

The aim of the present study was nanoencapsulation of thymol to improve its poor water solubility and preservation of encapsulated thymol against environmental conditions. Another goal of the current investigation was to assess the antibacterial activity of thymol nanoemulsion as a sustainable biopesticide to control the bacterial blight of cluster bean. An oil-in-water (o/w) nanoemulsion containing thymol was prepared by a high-energy emulsification method using gum acacia and soya lecithin as natural emulsifiers/surfactants. The characterization of thymol nanoemulsion was carried out using dynamic light scattering (DLS), transmission electron microscope (TEM) and Fourier transform infrared spectroscopy (FTIR). A mean particle size of about 83.38 nm was recorded within 10 min of sonication. The stability analysis of optimized nanoemulsion showed kinetic stability up to two months of storage at room temperature. The thymol nanoemulsion was found to be spherical with a size ranging from 80–200 nm in diameter using transmission electron microscopy. Fourier transform infrared spectroscopy was used to study the molecular interaction between emulsifier/surfactant and thymol. The antibacterial studies of thymol nanoemulsion (0.01–0.06%, v/v) by growth inhibition analysis showed a potential antibacterial effect against Xanthomonas axonopodis pv. cyamopsidis (18–0.1 log CFU/ml). Further, in field experiments, foliar spray of the different concentration of thymol nanoemulsion (0.01–0.06%, v/v) significantly increased the percent efficiency of disease control (25.06–94.48%) and reduced the disease intensity (67.33–4.25%) of bacterial blight in cluster bean.

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References

  1. Bagri RK (2021) Integrated pest management of cluster bean (Cyamopsis tetragonoloba L.). https://doi.org/10.22271/tpi.2021.v10.i3j.5855

  2. Kaur B, Purkayastha S, Dilbaghi N, Chaudhury A (2005) Characterization of Xanthomonas axonopodis pv. cyamopsidis, the bacterial blight pathogen of cluster bean using PCR-based molecular markers. J Phytopathol 153:470–479

    Article  CAS  Google Scholar 

  3. Kumhar DR, Meena AK, Meena PN (2018) Efficacy of different management modules against bacterial blight of cluster bean under epiphytotic conditions. J Pharmacogn Phytochem 7:1505–1509

    CAS  Google Scholar 

  4. Godara SL, Singh N (2021) Management of bacterial leaf blight (Xanthomonas axonopodis pv. cymopsidis) of cluster bean in Rajasthan. Indian Phytopathol. 74:223–227. https://doi.org/10.1007/s42360-020-00299-8

    Article  Google Scholar 

  5. Mishra S, Keswani C, Abhilash PC, Fraceto LF, Singh HB (2017) Integrated approach of agri-nanotechnology: challenges and future trends. Front Plant Sci 8:471. https://doi.org/10.3389/fpls.2017.00471

    Article  PubMed  PubMed Central  Google Scholar 

  6. Lade BD, Gogle DP, Lade DB, Moon GM, Nandeshwar SB, Kumbhare SD (2019) Nanobiopesticide formulations: application strategies today and future perspectives. In: Nano-biopesticides today and future perspectives. Academic Press, pp 179–206. https://doi.org/10.1016/B978-0-12-815829-6.00007-3

  7. Mishra S, Singh A, Keswani C, Singh HB (2014) Nanotechnology: exploring potential application in agriculture and its opportunities and constraints. Biotech today 4:9–14. https://doi.org/10.5958/2322-0996.2014.00011.8

    Article  Google Scholar 

  8. Kookana RS, Boxall AB, Reeves PT, Ashauer R, Beulke S, Chaudhry Q, Cornelis G, Fernandes TF, Gan J, Kah M, Lynch I (2014) Nanopesticides: guiding principles for regulatory evaluation of environmental risks. J Agric Food Chem 62:4227–4240. https://doi.org/10.1021/jf500232f

    Article  CAS  PubMed  Google Scholar 

  9. Nuruzzaman MD, Rahman MM, Liu Y, Naidu R (2016) Nanoencapsulation, nano-guard for pesticides: a new window for safe application. J Agric Food Chem 64:1447–1483. https://doi.org/10.1021/acs.jafc.5b05214

    Article  CAS  PubMed  Google Scholar 

  10. Pingali PL (2012) Green revolution: impacts, limits, and the path ahead. Proc Natl Acad Sci 109:12302–12308. https://doi.org/10.1073/pnas.0912953109

    Article  PubMed  PubMed Central  Google Scholar 

  11. Sedaghat Doost A, Devlieghere F, Dirckx A, Van der Meeren P (2018) Fabrication of Origanum compactum essential oil nanoemulsions stabilized using Quillaja Saponin biosurfactant. J Food Process Preserv 42:e13668. https://doi.org/10.1111/jfpp.13668

    Article  CAS  Google Scholar 

  12. De Castro RD, de Souza TMPA, Bezerra LMD, Ferreira GLS, de Brito Costa EMM, Cavalcanti AL (2015) Antifungal activity and mode of action of thymol and its synergism with nystatin against Candida species involved with infections in the oral cavity: an in vitro study. BMC Complement Altern Med 15:1–7. https://doi.org/10.1186/s12906-015-0947-2

    Article  CAS  Google Scholar 

  13. Chang Y, McLandsborough L, McClements DJ (2012) Physical properties and antimicrobial efficacy of thyme oil nanoemulsions: influence of ripening inhibitors. J Agric Food Chem 60:12056–12063. https://doi.org/10.1021/jf304045a

    Article  CAS  PubMed  Google Scholar 

  14. Kumari S, Kumaraswamy RV, Choudhary RC, Sharma SS, Pal A, Raliya R, Biswas P, Saharan V (2018) Thymol nanoemulsion exhibits potential antibacterial activity against bacterial pustule disease and growth promotory effect on soybean. Sci Rep 8:1–12. https://doi.org/10.1038/s41598-018-24871-5

    Article  CAS  Google Scholar 

  15. Chang Y, McLandsborough L, McClements DJ (2015) Fabrication, stability and efficacy of dual-component antimicrobial nanoemulsions: essential oil (thyme oil) and cationic surfactant (lauric arginate). Food Chem 172:298–304. https://doi.org/10.1016/j.foodchem.2014.09.081

    Article  CAS  PubMed  Google Scholar 

  16. Luo Y, Zhang Y, Pan K, Critzer F, Davidson PM, Zhong Q (2014) Self-emulsification of alkaline-dissolved clove bud oil by whey protein, gum arabic, lecithin, and their combinations. J Agric Food Chem 62:4417–4424. https://doi.org/10.1021/jf500698k

    Article  CAS  PubMed  Google Scholar 

  17. Liu LM, Huang SW, Wang L, Hou EQ, Xiao DF (2014) First report of leaf blight of rice caused by Cochliobolus lunatus in China. Plant Dis 98:686–686. https://doi.org/10.1094/PDIS-03-13-0303-PDN

    Article  CAS  PubMed  Google Scholar 

  18. Ghosh V, Mukherjee A, Chandrasekaran N (2014) Eugenol-loaded antimicrobial nanoemulsion preserves fruit juice against, microbial spoilage. Colloids Surf B 114:392–397. https://doi.org/10.1016/j.colsurfb.2013.10.034

    Article  CAS  Google Scholar 

  19. Esmaeili F, Rajabnejhad S, Partoazar AR, Mehr SE, Faridi-Majidi R, Sahebgharani M, Syedmoradi L, Rajabnejhad MR, Amani A (2016) Anti-inflammatory effects of eugenol nanoemulsion as a topical delivery system. Pharm Dev Technol 21:887–893. https://doi.org/10.3109/10837450.2015.1078353

    Article  CAS  PubMed  Google Scholar 

  20. Arakha M, Pal S, Samantarrai D, Panigrahi TK, Mallick BC, Pramanik K, Mallick B, Jha S (2015) Antimicrobial activity of iron oxide nanoparticle upon modulation of nanoparticle-bacteria interface. Sci Rep 5:1–12. https://doi.org/10.1038/srep14813

    Article  CAS  Google Scholar 

  21. Kim KH, Park JH, Kim MY, Heu S, Lee SH (2011) Genetic mapping of novel symptom in response to soybean bacterial leaf pustule in PI 96188. J Crop Sci Biotechnol 14:119–123. https://doi.org/10.1007/s12892-011-0024-4

    Article  Google Scholar 

  22. Sain SK, Gour HN (2009) Efficacy of isolated bacteria in vitro inhibition of Xanthomonas axonopodis pv. cyamopsidis and prevention of bacterial leaf blight of cluster bean in the field. J Biol Control 23:421–425. https://doi.org/10.18311/jbc/2009/3698

    Article  Google Scholar 

  23. Chester KS (1959) How sick is the plant? Plant Pathol 1:99–142

    Article  Google Scholar 

  24. Wheeler BEJ (1969) An introduction to plant diseases. An introduction to plant diseases

  25. Zhang J, Bing L, Reineccius GA (2016) Comparison of modified starch and Quillaja saponins in the formation and stabilization of flavor nanoemulsions. Food Chem 192:53–59. https://doi.org/10.1016/j.foodchem.2015.06.078

    Article  CAS  PubMed  Google Scholar 

  26. Kumari S, Choudhary RC, Kumaraswamy RV, Bhagat D, Pal A, Raliya R, Biswas P, Saharan V (2019) Zinc-functionalized thymol nanoemulsion for promoting soybean yield. Plant Physiol Biochem 145:64–74. https://doi.org/10.1016/j.plaphy.2019.10.022

    Article  CAS  PubMed  Google Scholar 

  27. Celebioglu A, Uyar T (2021) Electrohydrodynamic encapsulation of eugenol-cyclodextrin complexes in pullulan nanofibers. Food Hydrocolloids 111:106264. https://doi.org/10.1016/j.foodhyd.2020.106264

    Article  CAS  Google Scholar 

  28. Hu Z, Li S, Wang S, Zhang B, Huang Q (2021) Encapsulation of menthol into cyclodextrin metal-organic frameworks: preparation, structure characterization and evaluation of complexing capacity. Food Chem 338:127839. https://doi.org/10.1016/j.foodchem.2020.127839

    Article  CAS  PubMed  Google Scholar 

  29. Hu Q, Gerhard H, Upadhyaya I, Venkitanarayanan K, Luo Y (2016) Antimicrobial eugenol nanoemulsion prepared by gum arabic and lecithin and evaluation of drying technologies. Int J Biol Macromol 87:130–140. https://doi.org/10.1016/j.ijbiomac.2016.02.051

    Article  CAS  PubMed  Google Scholar 

  30. Ahmadi O, Jafarizadeh-Malmiri H (2021) Intensification process in thyme essential oil nanoemulsion preparation based on subcritical water as a green solvent and six different emulsifiers. Green Process Synth 10:430–439. https://doi.org/10.1515/gps-2021-0040

    Article  CAS  Google Scholar 

  31. Salvia-Trujillo L, Rojas-Graü A, Soliva-Fortuny R, Martín-Belloso O (2015) Physicochemical characterization and antimicrobial activity of food-grade emulsions and nanoemulsions incorporating essential oils. Food Hydrocolloids 43:547–556. https://doi.org/10.1016/j.foodhyd.2014.07.012

    Article  CAS  Google Scholar 

  32. Wu WH, Eskin DG, Priyadarshi A, Subroto T, Tzanakis I, Zhai W (2021) New insights into the mechanisms of ultrasonic emulsification in the oil–water system and the role of gas bubbles. Ultrason Sonochem 73:105501. https://doi.org/10.1016/j.ultsonch.2021.105501

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Moghimi R, Ghaderi L, Rafati H, Aliahmadi A, McClements DJ (2016) Superior antibacterial activity of nanoemulsion of Thymus daenensis essential oil against E. coli. Food Chem 194:410–415. https://doi.org/10.1016/j.foodchem.2015.07.139

    Article  CAS  PubMed  Google Scholar 

  34. Su D, Zhong Q (2016) Formation of thymol nanoemulsions with combinations of casein hydrolysates and sucrose stearate. J Food Eng 179:1–10. https://doi.org/10.1016/j.jfoodeng.2016.01.030

    Article  CAS  Google Scholar 

  35. Chemat F, Grondin I, Costes P, Moutoussamy L, Sing ASC, Smadja J (2004) High power ultrasound effects on lipid oxidation of refined sunflower oil. Ultrason Sonochem 11:281–285. https://doi.org/10.1016/j.ultsonch.2003.07.004

    Article  CAS  PubMed  Google Scholar 

  36. Ziani K, Chang Y, McLandsborough L, McClements DJ (2011) Influence of surfactant charge on antimicrobial efficacy of surfactant-stabilized thyme oil nanoemulsions. J Agric Food Chem 59:6247–6255. https://doi.org/10.1021/jf200450m

    Article  CAS  PubMed  Google Scholar 

  37. Gupta A, Eral HB, Hatton TA, Doyle PS (2016) Nanoemulsions: formation, properties and applications. Soft Matter 12:2826–2841. https://doi.org/10.1039/C5SM02958A

    Article  CAS  PubMed  Google Scholar 

  38. McClements DJ (2002) Colloidal basis of emulsion colour. Curr Opin Colloid Interface Sci 7:451–455. https://doi.org/10.1016/S1359-0294(02)00075-4

    Article  CAS  Google Scholar 

  39. Gupta P, Preet S, Ananya, Singh N (2022) Preparation of Thymus vulgaris (L.) essential oil nanoemulsion and its chitosan encapsulation for controlling mosquito vectors. Sci Rep 12:4335. https://doi.org/10.1038/s41598-022-07676-5

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Lazar-Baker EE, Hetherington SD, Ku VV, Newman SM (2011) Evaluation of commercial essential oil samples on the growth of postharvest pathogen Monilinia fructicola (G. Winter) Honey. Lett Appl Microbiol 52:227–232. https://doi.org/10.1111/j.1472-765X.2010.02996.x

    Article  CAS  PubMed  Google Scholar 

  41. Xue J, Davidson PM, Zhong Q (2017) Inhibition of Escherichia coli O157: H7 and Listeria monocytognes growth in milk and cantaloupe juice by thymol nanoemulsions prepared with gelatin and lecithin. Food Control 73:1499–1506. https://doi.org/10.1016/j.foodcont.2016.11.015

    Article  CAS  Google Scholar 

  42. Xu J, Zhou F, Ji BP, Pei RS, Xu N (2008) The antibacterial mechanism of carvacrol and thymol against Escherichia coli. Lett Appl Microbiol 47:174–179. https://doi.org/10.1111/j.1472-765X.2008.02407.x

    Article  CAS  PubMed  Google Scholar 

  43. Ben-Jabeur M, Ghabri E, Myriam M, Hamada W (2015) Thyme essential oil as a defense inducer of tomato against gray mold and Fusarium wilt. Plant Physiol Biochem 94:35–40. https://doi.org/10.1016/j.plaphy.2015.05.006

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

The authors are grateful to SAIF, AIIMS (New Delhi) for the TEM facility and the authors are also grateful to the Central instrumentation library (G.J.U S&T, Hisar) for the FTIR facility.

Funding

Pooja Choudhary is grateful to the Council of Scientific and Industrial Research (CSIR), New Delhi, [Ref. No. 09/752(0083)/2018/EMR-1] for providing financial support to her in the form of a Junior Research Fellowship (JRF). Dr. Gaurav is also thankful to CSIR, Govt. of India for providing CSIR-SRA (No. B-12998 dated 31 March 2023).

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Authors and Affiliations

  1. Department of Bio & Nano Technology, Guru Jambheshwar University of Science & Technology, Hisar, Haryana, 125001, India

    Pooja Choudhary, Gaurav Bhanjana, Sandeep Kumar & Neeraj Dilbaghi

  2. Physics Department, Punjab Engineering College (Deemed to be University), Chandigarh, 160012, India

    Sandeep Kumar

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  1. Pooja Choudhary
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  2. Gaurav Bhanjana
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Contributions

Pooja Choudhary: Data curation, Formal analysis, Investigation, Methodology, Validation, Visualization, Writing – original draft, Writing – review & editing; Gaurav Bhanjana: Review and Editing, Data Analysis; Neeraj Dilbaghi: Conceptualization, Data curation, Investigation, Methodology, Supervision, Writing – review & editing; Sandeep Kumar: Resources, Writing – review & editing.

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Correspondence to Neeraj Dilbaghi.

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Choudhary, P., Bhanjana, G., Kumar, S. et al. Antibacterial Activity of Sustainable Thymol Nanoemulsion Formulations Against the Bacterial Blight Disease on Cluster Bean Caused by Xanthomonas axonopodis. Indian J Microbiol (2024). https://doi.org/10.1007/s12088-024-01256-z

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