{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,17]],"date-time":"2026-06-17T15:21:33Z","timestamp":1781709693355,"version":"3.54.5"},"reference-count":56,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2021,5,10]],"date-time":"2021-05-10T00:00:00Z","timestamp":1620604800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005642","name":"University of Sassari","doi-asserted-by":"publisher","award":["FAR2020SATTA"],"award-info":[{"award-number":["FAR2020SATTA"]}],"id":[{"id":"10.13039\/501100005642","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Veterinary Sciences"],"abstract":"<jats:p>The use of natural substances such as essentials oils against bee pathogens is of great interest as an alternative to traditional methods based on synthetic compounds like antibiotics and fungicides, in order to minimize the risk of having toxic residues in hive products and to prevent the development of resistance phenomena. This study evaluated the inhibitory, fungicidal and sporicidal activity of ten essential oils extracted from aromatic plants against Ascosphaera apis, the etiological agent of chalkbrood, an invasive honey bee mycosis. The most effective essential oils were Thymus herba-barona, Thymus capitatus and Cinnamomum zeylanicum, which showed values of minimum fungicidal concentration and minimum sporicidal concentration ranging from 200 to 400 ppm. Carvacrol was the main component of Thymus capitatus and Thymus herba-barona oils, whereas cinnamic aldehyde prevailed in Cinnamomum\u00a0zeylanicum oil. Further in-apiary studies will allow the evaluation of side effects on bees and residues in hive products.<\/jats:p>","DOI":"10.3390\/vetsci8050080","type":"journal-article","created":{"date-parts":[[2021,5,10]],"date-time":"2021-05-10T10:49:51Z","timestamp":1620643791000},"page":"80","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["In Vitro Activity of Several Essential Oils Extracted from Aromatic Plants against Ascosphaera apis"],"prefix":"10.3390","volume":"8","author":[{"given":"Michelina","family":"Pusceddu","sequence":"first","affiliation":[{"name":"Department of Agricultural Sciences, University of Sassari, 07100 Sassari, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1274-7147","authenticated-orcid":false,"given":"Ignazio","family":"Floris","sequence":"additional","affiliation":[{"name":"Department of Agricultural Sciences, University of Sassari, 07100 Sassari, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0619-462X","authenticated-orcid":false,"given":"Nicoletta P.","family":"Mangia","sequence":"additional","affiliation":[{"name":"Department of Agricultural Sciences, University of Sassari, 07100 Sassari, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7370-0903","authenticated-orcid":false,"given":"Alberto","family":"Angioni","sequence":"additional","affiliation":[{"name":"Department of Life and Environmental Sciences, University of Cagliari, 09124 Cagliari, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1296-3835","authenticated-orcid":false,"given":"Alberto","family":"Satta","sequence":"additional","affiliation":[{"name":"Department of Agricultural Sciences, University of Sassari, 07100 Sassari, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,5,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"797","DOI":"10.1080\/00275514.1996.12026717","article-title":"Ascosphaera acerosa sp. nov. isolated from the alfalfa leafcutting bee, with a key to the species of Ascosphaera","volume":"88","author":"Bissett","year":"1996","journal-title":"Mycologia"},{"key":"ref_2","first-page":"7","article-title":"Ascosphaera torchioi sp. nov., a pathogen of Osmia lignaria propinqua (Hymenoptera)","volume":"77","author":"Youssef","year":"2001","journal-title":"Mycotaxon"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"501","DOI":"10.1002\/j.1537-2197.1955.tb11154.x","article-title":"Life cycle of Ascosphaera apis (Pericystis apis)","volume":"42","author":"Spiltoir","year":"1955","journal-title":"Am. J. Bot."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"553","DOI":"10.1080\/15572536.2007.11832549","article-title":"High mobility group (HMG-box) genes in the honeybee fungal pathogen Ascosphaera apis","volume":"99","author":"Aronstein","year":"2007","journal-title":"Mycologia"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1001","DOI":"10.1016\/j.sjbs.2016.04.016","article-title":"In vitro evaluation of the effects of some plant essential oils on Ascosphaera apis, the causative agent of Chalkbrood disease","volume":"24","author":"Ansari","year":"2017","journal-title":"Saudi J. Biol. Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1080\/00218839.1985.11100675","article-title":"Electropositive redox state of the fifth-instar larval gut of Apis mellifera","volume":"24","author":"Bignell","year":"1985","journal-title":"J. Apic. Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"540","DOI":"10.1051\/apido:19940604","article-title":"Chalkbrood development in honeybee brood under controlled conditions","volume":"25","author":"Puerta","year":"1994","journal-title":"Apidologie"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1051\/apido:19960401","article-title":"Effect of temperature and humidity of sealed brood on chalkbrood development under controlled conditions","volume":"27","author":"Flores","year":"1996","journal-title":"Apidologie"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1080\/00218839.1987.11100768","article-title":"Carbon dioxide activation of spores of the chalkbrood fungus Ascosphaera apis","volume":"26","author":"Heath","year":"1987","journal-title":"J. Apic. Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1080\/0005772X.1982.11097877","article-title":"Chalk brood pathogens: A review","volume":"63","author":"Heath","year":"1982","journal-title":"Bee World"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Bailey, L., and Ball, B.V. (1991). Honey Bee Pathology, Academic Press. [2nd ed.].","DOI":"10.1016\/B978-0-12-073481-8.50006-0"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"236","DOI":"10.3896\/IBRA.1.49.3.02","article-title":"In vitro inhibitory actions of some essential oils on Ascosphaera apis, a fungus responsible for honey bee chalkbrood","volume":"49","author":"Boudegga","year":"2010","journal-title":"J. Apic. Res."},{"key":"ref_13","first-page":"23","article-title":"The effect of Varroa jacobsoni Oud. on the incidence and course of chalkbrood disease in Apis mellifera L. colonies","volume":"43","author":"Glinski","year":"1988","journal-title":"Ann. Univ. Mariae Curie-Sk\u0142odowska Sect. DD Med. Vet."},{"key":"ref_14","first-page":"517","article-title":"Examination of artificially infested brood with Varroa mites for secondary infections","volume":"20","author":"Koch","year":"1989","journal-title":"Apidologie"},{"key":"ref_15","unstructured":"Needham, G.R., Page, E., Delfinado-Baker, M., and Bowman, C.E. (1988). Morphology of some microorganisms associated with the female mite Varroa jacobsoni: A survey by electron microscopy. Africanized Honey Bees and Bee Mites, Ellis Horwood Ltd."},{"key":"ref_16","first-page":"54","article-title":"Enfermedades secundarias a la parasitaci\u00f3n por Varroa en Apis mellifera","volume":"43","author":"Puerta","year":"1990","journal-title":"Vida Apic."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1016\/j.vetmic.2005.03.005","article-title":"Spores of Ascosphaera apis contained in wax foundation can infect honeybee brood","volume":"108","author":"Flores","year":"2005","journal-title":"Vet. Microbiol."},{"key":"ref_18","first-page":"703","article-title":"Assessment of losses in honey yield due to the chalkbrood disease, with reference to the determination of its economic injury levels in Egypt","volume":"70","author":"Zaghloul","year":"2005","journal-title":"Commun. Agri. Appl. Biol. Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"S20","DOI":"10.1016\/j.jip.2009.06.018","article-title":"Chalkbrood disease in honey bees","volume":"103","author":"Aronstein","year":"2010","journal-title":"J. Invertebr. Pathol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1007\/BF00441261","article-title":"Effects of itraconazole on the sporocysts wall of the entomopathogenic fungi Ascosphaera apis as revealed by the scanning electron microscope","volume":"103","author":"Liu","year":"1988","journal-title":"Mycopathologia"},{"key":"ref_21","first-page":"163","article-title":"Imidazole derivatives in control of the honey bee brood mycoses","volume":"40","author":"Glinski","year":"1996","journal-title":"Pszczel. Zesz. Nauk."},{"key":"ref_22","unstructured":"Davis, C., and Ward, W. (2003). Control of Chalkbrood Disease with Natural Products, Rural Industries Research and Development Corporation, Canprint. RIRDC Publication No 03\/2017."},{"key":"ref_23","first-page":"253","article-title":"In vitro inhibitory activity of essential oil vapors against Ascosphaera apis","volume":"7","author":"Kloucek","year":"2012","journal-title":"Nat. Prod. Commun."},{"key":"ref_24","first-page":"601","article-title":"A novel approach for the management of the chalkbrood disease infesting honeybee Apis mellifera L. (Hymenoptera: Apidae) colonies in Egypt","volume":"70","author":"Mourad","year":"2005","journal-title":"Commun. Agric. Appl. Biol. Sci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"704","DOI":"10.1080\/10412905.2006.9699211","article-title":"Laboratory evaluation of Heterothalamus alienus essential oil against different pests of Apis mellifera","volume":"18","author":"Ruffinengo","year":"2006","journal-title":"J. Essent. Oil Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1080\/10412905.2001.9699633","article-title":"In vitro fungistatic effect of essential oils against Ascosphaera apis","volume":"13","author":"Ringuelet","year":"2001","journal-title":"J. Essent. Oil Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"282","DOI":"10.1080\/10412905.2003.9712143","article-title":"In Vitro activity of essential oils from San Luis-Argentina against Ascosphaera apis","volume":"15","author":"Dellacasa","year":"2003","journal-title":"J. Essent. Oil Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"336","DOI":"10.1080\/10412905.2005.9698924","article-title":"An in vitro evaluation of Tagetes minuta essential oil for the control of the honeybee pathogens Paenibacillus larvae and Ascosphaera apis, and the parasitic mite Varroa destructor","volume":"17","author":"Eguaras","year":"2005","journal-title":"J. Essent. Oil Res."},{"key":"ref_29","first-page":"1","article-title":"Control of Ascosphaera apis and Paenibacillus larvae subsp. larvae by the use of essential oils for obtaining beehive products without toxic residues","volume":"11","author":"Bailac","year":"2006","journal-title":"Mol. Med. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"921","DOI":"10.1007\/s11046-018-0298-x","article-title":"Antimicrobial activity of essential oils against the fungal pathogens Ascosphaera apis and Pseudogymnoascus destructans","volume":"183","author":"Gabriel","year":"2018","journal-title":"Mycopathologia"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1080\/14786419.2017.1306703","article-title":"Stonebrood and chalkbrood in Apis mellifera causing fungi: In Vitro sensitivity to some essential oils","volume":"32","author":"Nardoni","year":"2018","journal-title":"Nat. Prod. Res."},{"key":"ref_32","first-page":"1147","article-title":"Factors affecting chemical variability of essential oils: A review of recent developments","volume":"4","author":"Barra","year":"2009","journal-title":"Nat. Prod. Commun."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1030","DOI":"10.1021\/jf025940c","article-title":"Chemical Composition, Plant Genetic Differences, and Antifungal Activity of the Essential Oil of Helichrysum italicum G. Don ssp. microphyllum (Willd) Nym","volume":"51","author":"Angioni","year":"2003","journal-title":"J. Agric. Food Chem."},{"key":"ref_34","unstructured":"Clinical and Laboratory Standards Institute (2008). Reference Method for Broth Dilution Antifungal Susceptibility Testing of Filamentous Fungi, Clinical and Laboratory Standards Institute. [2nd ed.]. Approved Standard."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1051\/apido:19960206","article-title":"Activit\u00e9s in vitro de plusieurs huiles essentielles sur Bacillus larvae White et essai au rucher","volume":"27","author":"Floris","year":"1996","journal-title":"Apidologie"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1085","DOI":"10.1007\/s00203-019-01673-5","article-title":"Antifungal activity of selected essential oils against Fusarium culmorum and F. graminearum and their secondary metabolites in wheat seeds","volume":"201","author":"Perczak","year":"2019","journal-title":"Arch. Microbiol."},{"key":"ref_37","unstructured":"R Core Team (2021, February 15). R: A Language and Environment for Statistical Computing. R Foundation for Statistical Computing. Available online: http:\/\/www.R-project.org."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1051\/apido:19890304","article-title":"Activit\u00e9 des huiles essentielles de Labi\u00e9es sur Ascophaera apis et traitement d\u2019un rucher","volume":"20","author":"Colin","year":"1989","journal-title":"Apidologie"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1603\/0022-0493-97.2.187","article-title":"Comparison between two thymol formulations in the control of Varroa destructor: Effectiveness, persistence, and residues","volume":"97","author":"Floris","year":"2004","journal-title":"J. Econ. Entomol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"4934","DOI":"10.1007\/s11356-013-2143-6","article-title":"Thymol as an alternative to pesticides: Persistence and effects of Apilife Var on the phototactic behavior of the honeybee Apis mellifera","volume":"21","author":"Carayon","year":"2014","journal-title":"Environ. Sci. Pollut. Res. Int."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"856","DOI":"10.1007\/s10646-016-1642-x","article-title":"Prolonged effects of in-hive monoterpenoids on the honey bee Apis mellifera","volume":"25","author":"Alayrangues","year":"2016","journal-title":"Ecotoxicology"},{"key":"ref_42","first-page":"1","article-title":"Antimicrobial activity of cinnamon (Cinnamomum zeylanicum) essential oil and its main components against Paenibacillus larvae from Argentine","volume":"61","author":"Gende","year":"2008","journal-title":"Bull. Insectology"},{"key":"ref_43","first-page":"93","article-title":"Advances in the apiary control of the honeybee American Foulbrood with Cinnamon (Cinnamomum zeylanicum) essential oil","volume":"62","author":"Gende","year":"2009","journal-title":"Bull. Insectology"},{"key":"ref_44","first-page":"247","article-title":"Susceptibility of Paenibacillus larvae isolates to a tetracycline hydrochloride and Cinnamon (Cinnamomum zeylanicum) essential oil mixture","volume":"63","author":"Gende","year":"2010","journal-title":"Bull. Insectology"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1080\/10412905.1994.9698376","article-title":"An in vitro evaluation of botanical compounds for the control of the honeybee pathogens Bacillus larvae and Ascosphaera apis, and the secondary invader B. alvei","volume":"6","author":"Calderone","year":"1994","journal-title":"J. Essent. Oil Res."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"704","DOI":"10.1038\/194704b0","article-title":"Identification of geraniol as the active component in the Nassanoff pheromone of the honey bee","volume":"194","author":"Boch","year":"1962","journal-title":"Nature"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"320","DOI":"10.1038\/202320a0","article-title":"Identification of nerolic and geranic acids in the Nassanoff pheromone of the honey bee","volume":"202","author":"Boch","year":"1964","journal-title":"Nature"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1513","DOI":"10.1016\/0022-1910(66)90041-2","article-title":"Citral in the Nassanoff pheromone of the honey bee","volume":"12","author":"Shearer","year":"1966","journal-title":"J. Insect Physiol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1016\/j.fbio.2019.04.004","article-title":"In vitro antibacterial activity and volatile characterisation of organic Apis mellifera ligustica (Spinola, 1906) beeswax ethanol extracts","volume":"29","author":"Felicioli","year":"2019","journal-title":"Food Biosci."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"9514","DOI":"10.3390\/ijms12129514","article-title":"Volatile compounds in honey: A review on their involvement in aroma, botanical origin determination and potential biomedical activities","volume":"12","author":"Ndip","year":"2011","journal-title":"Int. J. Mol. Sci."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1051\/apido:19950202","article-title":"Propolis: Chemical composition, biological properties and therapeutic activity","volume":"26","author":"Marcucci","year":"1995","journal-title":"Apidologie"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1051\/apido:2000102","article-title":"Propolis: Recent advances in chemistry and plant origin","volume":"31","author":"Bankova","year":"2000","journal-title":"Apidologie"},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Simone-Finstrom, M.D., and Spivak, M. (2012). Increased resin collection after parasite challenge: A case of self-medication in honey bees?. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0034601"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"297","DOI":"10.1111\/1744-7917.12515","article-title":"Resin foraging dynamics in Varroa destructor-infested hives: A case of medication of kin?","volume":"26","author":"Pusceddu","year":"2019","journal-title":"Insect Sci."},{"key":"ref_55","first-page":"317","article-title":"Botanicals in pest management: Current status and future perspectives","volume":"Volume 1","author":"Peshin","year":"2009","journal-title":"Integrated Pest Management: Innovation-Development Process"},{"key":"ref_56","first-page":"63","article-title":"Essential oils as green pesticides: Potential and constraints","volume":"4","author":"Koul","year":"2008","journal-title":"Biopestic Int."}],"container-title":["Veterinary Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2306-7381\/8\/5\/80\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:58:45Z","timestamp":1760162325000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2306-7381\/8\/5\/80"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,5,10]]},"references-count":56,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2021,5]]}},"alternative-id":["vetsci8050080"],"URL":"https:\/\/doi.org\/10.3390\/vetsci8050080","relation":{},"ISSN":["2306-7381"],"issn-type":[{"value":"2306-7381","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,5,10]]}}}