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However, these compounds have limitations such as volatility, poor water solubility, and phytotoxicity. Nanoencapsulation, through processes like micro- and nanoemulsions, can enhance the stability and bioactivity of essential oils. In this study, thyme essential oil from supercritical carbon dioxide extraction was selected as a sustainable antimicrobial tool and nanoencapsulated in an oil-in-water emulsion system. The investigated protocol provided high-speed homogenisation in the presence of cellulose nanocrystals as stabilisers and calcium chloride as an ionic crosslinking agent. Thyme essential oil was characterised via GC-MS and UV-vis analysis, indicating rich content in phenols. The cellulose nanocrystal\/essential oil ratio and calcium chloride concentration were varied to tune the nanoemulsions\u2019 physical\u2013chemical stability, which was investigated via UV-vis, direct observation, dynamic light scattering, and Turbiscan analysis. Transmission electron microscopy confirmed the nanosized droplet formation. The nanoemulsion resulting from the addition of crosslinked nanocrystals was very stable over time at room temperature. It was evaluated for the first time on Pseudomonas savastanoi pv. savastanoi, the causal agent of olive knot disease. In vitro tests showed a synergistic effect of the formulation components, and in vivo tests on olive seedlings demonstrated reduced bacterial colonies without any phytotoxic effect. These findings suggest that crosslinked cellulose nanocrystal emulsions can enhance the stability and bioactivity of thyme essential oil, providing a new tool for crop protection.<\/jats:p>","DOI":"10.3390\/molecules28237884","type":"journal-article","created":{"date-parts":[[2023,12,1]],"date-time":"2023-12-01T04:12:56Z","timestamp":1701403976000},"page":"7884","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Cellulose Nanocrystal-Based Emulsion of Thyme Essential Oil: Preparation and Characterisation as Sustainable Crop Protection Tool"],"prefix":"10.3390","volume":"28","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3328-7701","authenticated-orcid":false,"given":"Francesca","family":"Baldassarre","sequence":"first","affiliation":[{"name":"Department of Biological and Environmental Sciences, UdR INSTM of Lecce University of Salento, Via Monteroni, 73100 Lecce, Italy"},{"name":"Institute of Nanotechnology, CNR NANOTEC, Consiglio Nazionale delle Ricerche, Via Monteroni, 73100 Lecce, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2743-6413","authenticated-orcid":false,"given":"Daniele","family":"Schiavi","sequence":"additional","affiliation":[{"name":"Department of Agriculture and Forest Sciences (DAFNE), University of Tuscia, Via S. Camillo de Lellis, snc, 01100 Viterbo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Veronica","family":"Di Lorenzo","sequence":"additional","affiliation":[{"name":"Department of Agriculture and Forest Sciences (DAFNE), University of Tuscia, Via S. Camillo de Lellis, snc, 01100 Viterbo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesca","family":"Biondo","sequence":"additional","affiliation":[{"name":"Department of Biological and Environmental Sciences, UdR INSTM of Lecce University of Salento, Via Monteroni, 73100 Lecce, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3706-6276","authenticated-orcid":false,"given":"Viviana","family":"Vergaro","sequence":"additional","affiliation":[{"name":"Department of Biological and Environmental Sciences, UdR INSTM of Lecce University of Salento, Via Monteroni, 73100 Lecce, Italy"},{"name":"Institute of Nanotechnology, CNR NANOTEC, Consiglio Nazionale delle Ricerche, Via Monteroni, 73100 Lecce, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9182-4336","authenticated-orcid":false,"given":"Gianpiero","family":"Colangelo","sequence":"additional","affiliation":[{"name":"Department of Engineering for Innovation, University of Salento, Via Monteroni, 73100 Lecce, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0822-4506","authenticated-orcid":false,"given":"Giorgio Mariano","family":"Balestra","sequence":"additional","affiliation":[{"name":"Department of Agriculture and Forest Sciences (DAFNE), University of Tuscia, Via S. Camillo de Lellis, snc, 01100 Viterbo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4467-0922","authenticated-orcid":false,"given":"Giuseppe","family":"Ciccarella","sequence":"additional","affiliation":[{"name":"Department of Biological and Environmental Sciences, UdR INSTM of Lecce University of Salento, Via Monteroni, 73100 Lecce, Italy"},{"name":"Institute of Nanotechnology, CNR NANOTEC, Consiglio Nazionale delle Ricerche, Via Monteroni, 73100 Lecce, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,11,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/j.envint.2017.05.018","article-title":"Residential exposure to pesticides as risk factor for childhood and young adult brain tumors: A systematic review and meta-analysis","volume":"106","author":"Lison","year":"2017","journal-title":"Environ. 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