{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,28]],"date-time":"2026-03-28T19:09:28Z","timestamp":1774724968370,"version":"3.50.1"},"reference-count":86,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2022,10,17]],"date-time":"2022-10-17T00:00:00Z","timestamp":1665964800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"FCT\/MCTES (PIDDAC)","award":["LA\/P\/0045\/2020"],"award-info":[{"award-number":["LA\/P\/0045\/2020"]}]},{"name":"FCT\/MCTES (PIDDAC)","award":["UIDB\/00511\/2020"],"award-info":[{"award-number":["UIDB\/00511\/2020"]}]},{"name":"FCT\/MCTES (PIDDAC)","award":["UIDP\/00511\/2020"],"award-info":[{"award-number":["UIDP\/00511\/2020"]}]},{"name":"FCT\/MCTES (PIDDAC)","award":["NORTE-01-0145-FEDER-000054"],"award-info":[{"award-number":["NORTE-01-0145-FEDER-000054"]}]},{"name":"FCT\/MCTES (PIDDAC)","award":["UIDB\/04033\/2020"],"award-info":[{"award-number":["UIDB\/04033\/2020"]}]},{"name":"Norte Portugal Regional Operational Programme (NORTE 2020)","award":["LA\/P\/0045\/2020"],"award-info":[{"award-number":["LA\/P\/0045\/2020"]}]},{"name":"Norte Portugal Regional Operational Programme (NORTE 2020)","award":["UIDB\/00511\/2020"],"award-info":[{"award-number":["UIDB\/00511\/2020"]}]},{"name":"Norte Portugal Regional Operational Programme (NORTE 2020)","award":["UIDP\/00511\/2020"],"award-info":[{"award-number":["UIDP\/00511\/2020"]}]},{"name":"Norte Portugal Regional Operational Programme (NORTE 2020)","award":["NORTE-01-0145-FEDER-000054"],"award-info":[{"award-number":["NORTE-01-0145-FEDER-000054"]}]},{"name":"Norte Portugal Regional Operational Programme (NORTE 2020)","award":["UIDB\/04033\/2020"],"award-info":[{"award-number":["UIDB\/04033\/2020"]}]},{"name":"national funds of the FCT-Portuguese Foundation for Science and Technology","award":["LA\/P\/0045\/2020"],"award-info":[{"award-number":["LA\/P\/0045\/2020"]}]},{"name":"national funds of the FCT-Portuguese Foundation for Science and Technology","award":["UIDB\/00511\/2020"],"award-info":[{"award-number":["UIDB\/00511\/2020"]}]},{"name":"national funds of the FCT-Portuguese Foundation for Science and Technology","award":["UIDP\/00511\/2020"],"award-info":[{"award-number":["UIDP\/00511\/2020"]}]},{"name":"national funds of the FCT-Portuguese Foundation for Science and Technology","award":["NORTE-01-0145-FEDER-000054"],"award-info":[{"award-number":["NORTE-01-0145-FEDER-000054"]}]},{"name":"national funds of the FCT-Portuguese Foundation for Science and Technology","award":["UIDB\/04033\/2020"],"award-info":[{"award-number":["UIDB\/04033\/2020"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomedicines"],"abstract":"<jats:p>Essential oils are natural compounds used by humans for scientific purposes due to their wide range of properties. Eugenol is mostly present in clove oil, while pulegone is the main constituent of pennyroyal oil. To guarantee the safe use of eugenol and pulegone for both humans and animals, this study addressed, for the first time, the effects of these compounds, at low doses (chronic toxicity) and high doses (acute toxicity), in laboratory animals. Thirty-five FVB\/n female mice were randomly assigned to seven groups (n = 5): group I (control, non-additive diet); group II (2.6 mg of eugenol + 2.6 mg of pulegone); group III (5.2 mg of eugenol + 5.2 mg of pulegone); group IV (7.8 mg of eugenol + 7.8 mg of pulegone); group V (7.8 mg of eugenol); group VI (7.8 mg of pulegone); and group VII (1000 mg of eugenol + 1000 mg of pulegone). The compounds were administered in the food. Groups I to VI were integrated into the chronic toxicity study, lasting 28 days, and group VII was used in the acute toxicity study, lasting 7 days. Animals were monitored to assess their general welfare. Water and food intake, as well as body weight, were recorded. On the 29th day, all animals were euthanized by an overdose of ketamine and xylazine, and a complete necropsy was performed. Blood samples were collected directly from the heart for microhematocrit and serum analysis, as well as for comet assay. Organs were collected, weighed, and fixed in formaldehyde for further histological analysis and enzymatic assay. Eugenol and pulegone induced behavioral changes in the animals, namely in the posture, hair appearance and grooming, and in mental status. These compounds also caused a decrease in the animals\u2019 body weight, as well as in the food and water consumption. A mortality rate of 20% was registered in the acute toxicity group. Both compounds modulated the serum levels of triglycerides and alanine aminotransferase. Eugenol and pulegone induced genetic damage in all animals. Eugenol increased the activity of the CAT enzyme. Both compounds increased the GR enzyme at the highest dose. Moreover, pulegone administered as a single compound increased the activity of the GST enzyme. Histopathological analysis revealed inflammatory infiltrates in the lungs of groups II, III, and IV. The results suggest that eugenol and pulegone may exert beneficial or harmful effects, depending on the dose, and if applied alone or in combination.<\/jats:p>","DOI":"10.3390\/biomedicines10102595","type":"journal-article","created":{"date-parts":[[2022,10,17]],"date-time":"2022-10-17T05:08:02Z","timestamp":1665983282000},"page":"2595","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["Pulegone and Eugenol Oral Supplementation in Laboratory Animals: Results from Acute and Chronic Studies"],"prefix":"10.3390","volume":"10","author":[{"given":"Carla M.","family":"Ribeiro-Silva","sequence":"first","affiliation":[{"name":"School of Life and Environmental Sciences (ECVA), University of Tr\u00e1s-os-Montes and Alto Douro (UTAD), 5000-801 Vila Real, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5572-6317","authenticated-orcid":false,"given":"Ana I.","family":"Faustino-Rocha","sequence":"additional","affiliation":[{"name":"Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB) Inov4Agro, 5000-801 Vila Real, Portugal"},{"name":"Department of Zootechnics, School of Sciences and Technology (ECT), University of \u00c9vora, 7002-554 \u00c9vora, Portugal"},{"name":"Comprehensive Health Research Centre (CHRC), 7000-811 \u00c9vora, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2151-2449","authenticated-orcid":false,"given":"Rui M.","family":"Gil da Costa","sequence":"additional","affiliation":[{"name":"Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB) Inov4Agro, 5000-801 Vila Real, Portugal"},{"name":"Molecular Oncology and Viral Pathology Group, Research Center of IPO Porto (CI-IPOP)\/RISE@CI-IPOP (Health Research Network), Portuguese Oncology Institute of Porto (IPO Porto), Porto Comprehensive Cancer Center (Porto.CCC), 4200-072 Porto, Portugal"},{"name":"Laboratory for Process Engineering, Environment, Biotechnology and Energy (LEPABE), Faculty of Engineering, University of Porto (FEUP), 4200-465 Porto, Portugal"},{"name":"Associate Laboratory in Chemical Engineering (ALiCE), FEUP, 4200-465 Porto, Portugal"},{"name":"Postgraduate Programme in Adult Health (PPGSAD), Department of Morphology, Federal University of Maranh\u00e3o (UFMA), UFMA University Hospital (HUUFMA), S\u00e3o Lu\u00eds 65020-070, Brazil"}]},{"given":"Rui","family":"Medeiros","sequence":"additional","affiliation":[{"name":"Molecular Oncology and Viral Pathology Group, Research Center of IPO Porto (CI-IPOP)\/RISE@CI-IPOP (Health Research Network), Portuguese Oncology Institute of Porto (IPO Porto), Porto Comprehensive Cancer Center (Porto.CCC), 4200-072 Porto, Portugal"},{"name":"Research Department of the Portuguese League against Cancer Regional Nucleus of the North (LPCC-NRN), 4200-177 Porto, Portugal"},{"name":"Faculty of Medicine, University of Porto, 4200-319 Porto, Portugal"},{"name":"Institute of Biomedical Sciences Abel Salazar (ICBAS), 4050-313 Porto, Portugal"},{"name":"Biomedical Research Center (CEBIMED), Faculty of Health Sciences of Fernando Pessoa University (UFP), 4249-004 Porto, Portugal"}]},{"given":"Maria J.","family":"Pires","sequence":"additional","affiliation":[{"name":"Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB) Inov4Agro, 5000-801 Vila Real, Portugal"},{"name":"Department of Veterinary Sciences, UTAD, 5000-801 Vila Real, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4461-7021","authenticated-orcid":false,"given":"Isabel","family":"Gaiv\u00e3o","sequence":"additional","affiliation":[{"name":"School of Life and Environmental Sciences (ECVA), University of Tr\u00e1s-os-Montes and Alto Douro (UTAD), 5000-801 Vila Real, Portugal"},{"name":"Animal and Veterinary Research Centre (CECAV), Associate Laboratory for Animal and Veterinary Science\u2014AL4AnimalS, 5000-801 Vila Real, Portugal"},{"name":"Department of Genetics and Biotechnology, 5000-801 Vila Real, Portugal"}]},{"given":"Adelina","family":"Gama","sequence":"additional","affiliation":[{"name":"Department of Veterinary Sciences, UTAD, 5000-801 Vila Real, Portugal"},{"name":"Department of Genetics and Biotechnology, 5000-801 Vila Real, Portugal"},{"name":"Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), UTAD, 5000-801 Vila Real, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1464-8700","authenticated-orcid":false,"given":"Maria J.","family":"Neuparth","sequence":"additional","affiliation":[{"name":"Research Center in Physical Activity, Health and Leisure (CIAFEL), Faculty of Sports, O.Porto, University of Porto (FADEUP), 4200-319 Porto, Portugal"},{"name":"Laboratory for Integrative and Translational Research in Population Health (ITR), 4050-600 Porto, Portugal"},{"name":"Toxicology Research Unit (TOXRUN), University Institute of Health Sciences, CESPU, CRL, 4585-116 Gandra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8018-0317","authenticated-orcid":false,"given":"Joana V.","family":"Barbosa","sequence":"additional","affiliation":[{"name":"Laboratory for Process Engineering, Environment, Biotechnology and Energy (LEPABE), Faculty of Engineering, University of Porto (FEUP), 4200-465 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9729-7477","authenticated-orcid":false,"given":"Francisco","family":"Peixoto","sequence":"additional","affiliation":[{"name":"School of Life and Environmental Sciences (ECVA), University of Tr\u00e1s-os-Montes and Alto Douro (UTAD), 5000-801 Vila Real, Portugal"},{"name":"Chemistry Center\u2014Vila Real (CQ-VR), Biological and Environment Department, UTAD, 5000-801 Vila Real, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4447-4473","authenticated-orcid":false,"given":"Fern\u00e3o D.","family":"Magalh\u00e3es","sequence":"additional","affiliation":[{"name":"Laboratory for Process Engineering, Environment, Biotechnology and Energy (LEPABE), Faculty of Engineering, University of Porto (FEUP), 4200-465 Porto, Portugal"},{"name":"Associate Laboratory in Chemical Engineering (ALiCE), FEUP, 4200-465 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8605-0054","authenticated-orcid":false,"given":"Margarida M. S. M.","family":"Bastos","sequence":"additional","affiliation":[{"name":"Laboratory for Process Engineering, Environment, Biotechnology and Energy (LEPABE), Faculty of Engineering, University of Porto (FEUP), 4200-465 Porto, Portugal"},{"name":"Associate Laboratory in Chemical Engineering (ALiCE), FEUP, 4200-465 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9519-4044","authenticated-orcid":false,"given":"Paula A.","family":"Oliveira","sequence":"additional","affiliation":[{"name":"Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB) Inov4Agro, 5000-801 Vila Real, Portugal"},{"name":"Department of Veterinary Sciences, UTAD, 5000-801 Vila Real, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2022,10,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"372","DOI":"10.1016\/j.biortech.2009.07.048","article-title":"Repellent Activity of Essential Oils: A Review","volume":"101","author":"Nerio","year":"2010","journal-title":"Bioresour. 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