{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,2]],"date-time":"2026-04-02T20:08:11Z","timestamp":1775160491752,"version":"3.50.1"},"reference-count":54,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2020,6,12]],"date-time":"2020-06-12T00:00:00Z","timestamp":1591920000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Some evidence indicated that human babesiosis caused by Babesia duncani has spread widely in North America. However, current therapeutic regimens (atovaquone + azithromycin) for human babesiosis are suboptimal with frequent recrudescence and side effects, and furthermore, there is no specific treatment for human babesiosis caused by B. duncani. Here, we screened 97 essential oils and identified 10 essential oils (garlic, black pepper, tarragon, palo santo, coconut, pine, meditation, cajeput, moringa, and stress relief) at a low concentration (0.001%; v\/v) that showed good inhibitory activity against B. duncani in the hamster red blood cell culture model. Among them, garlic oil and black pepper oil performed best, as well as their potential active ingredients diallyl disulfide (DADS) and \u03b2-caryophyllene (BCP), respectively. Interestingly, further subculture study indicated that B. duncani could relapse after treatment with current therapeutic drugs atovaquone or azithromycin even at high concentrations. In contrast, the combination of garlic oil or DADS and azithromycin showed eradication of B. duncani at low concentrations without regrowth. These results are encouraging and suggest that the garlic-derived sulfur compound DADS and \u03b2-caryophyllene (BCP) may be promising drug candidates for evaluation of their ability to cure persistent B. duncani infections in the future.<\/jats:p>","DOI":"10.3390\/pathogens9060466","type":"journal-article","created":{"date-parts":[[2020,6,15]],"date-time":"2020-06-15T03:17:32Z","timestamp":1592191052000},"page":"466","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Identification of Essential Oils Including Garlic Oil and Black Pepper Oil with High Activity against Babesia duncani"],"prefix":"10.3390","volume":"9","author":[{"given":"Yumin","family":"Zhang","sequence":"first","affiliation":[{"name":"Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 21205, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chunxiang","family":"Bai","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 21205, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wanliang","family":"Shi","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 21205, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6118-1066","authenticated-orcid":false,"given":"Hector","family":"Alvarez-Manzo","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 21205, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5908-307X","authenticated-orcid":false,"given":"Ying","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 21205, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,6,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2397","DOI":"10.1056\/NEJMra1202018","article-title":"Human babesiosis","volume":"366","author":"Vannier","year":"2012","journal-title":"N. Engl. J. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1111\/j.1423-0410.2006.00740.x","article-title":"Babesiosis and blood transfusion: Flying under the radar","volume":"90","author":"Leiby","year":"2006","journal-title":"Vox Sang."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"116","DOI":"10.3201\/eid0901.020149","article-title":"Transfusion-associated babesiosis after heart transplant","volume":"9","author":"Lux","year":"2003","journal-title":"Emerg. Infect. Dis."},{"key":"ref_4","unstructured":"(2020, June 01). CDC - Babesiosis - Data & Statistics Data, Available online: https:\/\/www.cdc.gov\/parasites\/babesiosis\/data-statistics\/index.html."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"284","DOI":"10.7326\/0003-4819-119-4-199308150-00006","article-title":"Babesiosis in Washington State: A new species of Babesia?","volume":"119","author":"Quick","year":"1993","journal-title":"Ann. Intern. Med."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Scott, J.D., and Scott, C.M. (2018). Human Babesiosis Caused by Babesia duncani Has Widespread Distribution across Canada. Healthc. Basel, 6.","DOI":"10.3390\/healthcare6020049"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"779","DOI":"10.1016\/j.ijpara.2006.03.008","article-title":"Description of Babesia duncani n.sp. (Apicomplexa: Babesiidae) from humans and its differentiation from other piroplasms","volume":"36","author":"Conrad","year":"2006","journal-title":"Int. J. Parasitol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/j.ijpara.2018.05.008","article-title":"Insights into the evolution and drug susceptibility of Babesia duncani from the sequence of its mitochondrial and apicoplast genomes","volume":"49","author":"Virji","year":"2019","journal-title":"Int. J. Parasitol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1323","DOI":"10.1016\/S0020-7519(00)00137-5","article-title":"Human babesiosis: An emerging tick-borne disease","volume":"30","author":"Kjemtrup","year":"2000","journal-title":"Int. J. Parasitol."},{"key":"ref_10","unstructured":"(2020, June 01). CDC - Babesiosis - Resources for Health Professionals, Available online: https:\/\/www.cdc.gov\/parasites\/babesiosis\/health_professionals\/index.html#tx."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1056\/NEJM199807163390304","article-title":"Persistent parasitemia after acute babesiosis","volume":"339","author":"Krause","year":"1998","journal-title":"N. Engl. J. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1016\/j.diagmicrobio.2016.02.016","article-title":"Persistence of babesiosis for >2 years in a patient on rituximab for rheumatoid arthritis","volume":"85","author":"Raffalli","year":"2016","journal-title":"Diagn. Microbiol. Infect. Dis."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1050","DOI":"10.1093\/infdis\/169.5.1050","article-title":"Cultivation and phylogenetic characterization of a newly recognized human pathogenic protozoan","volume":"169","author":"Thomford","year":"1994","journal-title":"J. Infect. Dis."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"19974","DOI":"10.1074\/jbc.AC118.005771","article-title":"Establishment of a continuous in vitro culture of Babesia duncani in human erythrocytes reveals unusually high tolerance to recommended therapies","volume":"293","author":"Abraham","year":"2018","journal-title":"J. Biol. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"12","DOI":"10.3389\/fmicb.2012.00012","article-title":"Essential oils in food preservation: Mode of action, synergies, and interactions with food matrix components","volume":"3","author":"Hyldgaard","year":"2012","journal-title":"Front. Microbiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"680","DOI":"10.1002\/ptr.951","article-title":"Studies on essential oils: Part 10; antibacterial activity of volatile oils of some spices","volume":"16","author":"Singh","year":"2002","journal-title":"Phytother. Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"212","DOI":"10.5455\/jice.20160331064446","article-title":"In vitro antimicrobial activity of five essential oils on multidrug resistant Gram-negative clinical isolates","volume":"5","author":"Sakkas","year":"2016","journal-title":"J. Intercult. Ethnopharmacol."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Feng, J., Zhang, S., Shi, W.T., Zubcevik, N., Miklossy, J., and Zhang, Y. (2017). Selective Essential Oils from Spice or Culinary Herbs Have High Activity against Stationary Phase and Biofilm Borrelia burgdorferi. Front. Med. Lausanne, 4.","DOI":"10.3389\/fmed.2017.00169"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Ma, X., Shi, W., and Zhang, Y. (2019). Essential Oils with High Activity against Stationary Phase Bartonella henselae. Antibiotics (Basel), 8.","DOI":"10.20944\/preprints201910.0178.v1"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1080\/10412905.2003.9712099","article-title":"Essential oil composition of four major cultivars of black pepper (Piper nigrum L.) III","volume":"15","author":"Menon","year":"2003","journal-title":"J. Essent. Oil Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1","DOI":"10.15585\/mmwr.ss6806a1","article-title":"Babesiosis Surveillance\u2014United States, 2011\u20132015","volume":"68","author":"Gray","year":"2019","journal-title":"MMWR Surveill. Summ."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"509","DOI":"10.7326\/0003-4819-155-8-201110180-00362","article-title":"Transfusion-associated babesiosis in the United States: A description of cases","volume":"155","author":"Herwaldt","year":"2011","journal-title":"Ann. Intern. Med."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1128\/AEM.67.1.475-480.2001","article-title":"Antimicrobial properties of garlic oil against human enteric bacteria: Evaluation of methodologies and comparisons with garlic oil sulfides and garlic powder","volume":"67","author":"Ross","year":"2001","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/S0740-0020(03)00046-7","article-title":"Inhibitory effect of thyme and basil essential oils, carvacrol, thymol, estragol, linalool and p-cymene towards Shigella sonnei and S-flexneri","volume":"21","author":"Bagamboula","year":"2004","journal-title":"Food Microbiol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1007\/BF00383068","article-title":"A quantitative assessment of the antimicrobial activity of garlic (Allium sativum)","volume":"9","author":"Rees","year":"1993","journal-title":"World J. Microbiol. Biotechnol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"282","DOI":"10.1007\/s002530100722","article-title":"Antimicrobial properties of Allium sativum (garlic)","volume":"57","author":"Harris","year":"2001","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"499","DOI":"10.1128\/AAC.30.3.499","article-title":"Allium sativum (garlic) inhibits lipid synthesis by Candida albicans","volume":"30","author":"Adetumbi","year":"1986","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"417","DOI":"10.1055\/s-2006-961504","article-title":"In vitro virucidal effects of Allium sativum (garlic) extract and compounds","volume":"58","author":"Weber","year":"1992","journal-title":"Planta Med."},{"key":"ref_29","first-page":"256","article-title":"Antimycobacterial and Antibacterial Activity of Allium sativum Bulbs","volume":"76","author":"Viswanathan","year":"2014","journal-title":"Indian J. Pharm. Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"M390","DOI":"10.1111\/j.1750-3841.2009.01289.x","article-title":"Antibacterial effects of allspice, garlic, and oregano essential oils in tomato films determined by overlay and vapor-phase methods","volume":"74","author":"Du","year":"2009","journal-title":"J. Food Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1731","DOI":"10.1128\/AAC.50.5.1731-1737.2006","article-title":"Antimalarial activity of allicin, a biologically active compound from garlic cloves","volume":"50","author":"Coppi","year":"2006","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1093\/infdis\/156.1.243","article-title":"Inhibition of growth of Entamoeba histolytica by allicin, the active principle of garlic extract (Allium sativum)","volume":"156","author":"Mirelman","year":"1987","journal-title":"J. Infect. Dis."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1007\/s00436-013-3654-2","article-title":"Inhibitory effect of allicin on the growth of Babesia and Theileria equi parasites","volume":"113","author":"Salama","year":"2014","journal-title":"Parasitol. Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1002\/em.20474","article-title":"Biological properties of garlic and garlic-derived organosulfur compounds","volume":"50","author":"Iciek","year":"2009","journal-title":"Environ. Mol. Mutagen."},{"key":"ref_35","first-page":"2094","article-title":"Antibacterial Activity and Mechanism of Action of Black Pepper Essential Oil on Meat-Borne Escherichia coli","volume":"7","author":"Zhang","year":"2016","journal-title":"Front. Microbiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"17773","DOI":"10.3390\/molecules191117773","article-title":"Antibacterial and antioxidant activity of essential oil terpenes against pathogenic and spoilage-forming bacteria and cell structure-activity relationships evaluated by SEM microscopy","volume":"19","author":"Zengin","year":"2014","journal-title":"Molecules"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1007\/s00436-011-2265-z","article-title":"In vitro sensitivity of Blastocystis hominis to garlic, ginger, white cumin, and black pepper used in diet","volume":"109","author":"Yakoob","year":"2011","journal-title":"Parasitol. Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1307","DOI":"10.1084\/jem.20151519","article-title":"Radical cure of experimental babesiosis in immunodeficient mice using a combination of an endochin-like quinolone and atovaquone","volume":"213","author":"Lawres","year":"2016","journal-title":"J. Exp. Med."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"5059","DOI":"10.1128\/AAC.00928-16","article-title":"Extensive Shared Chemosensitivity between Malaria and Babesiosis Blood-Stage Parasites","volume":"60","author":"Paul","year":"2016","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1016\/S0140-6736(00)74210-3","article-title":"Treatment of staphylococcal infections with penicillin\u2014By intermittent sterilisation","volume":"2","author":"Bigger","year":"1944","journal-title":"Lancet"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"607","DOI":"10.1038\/s41579-019-0238-x","article-title":"Protozoan persister-like cells and drug treatment failure","volume":"17","author":"Barrett","year":"2019","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1016\/j.drup.2015.10.001","article-title":"Defining the frontiers between antifungal resistance, tolerance and the concept of persistence","volume":"23","author":"Delarze","year":"2015","journal-title":"Drug Resist. Update"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"370","DOI":"10.1086\/525852","article-title":"Persistent and relapsing babesiosis in immunocompromised patients","volume":"46","author":"Krause","year":"2008","journal-title":"Clin. Infect. Dis."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"428","DOI":"10.1128\/AAC.05456-11","article-title":"Phenotypic changes in artemisinin-resistant Plasmodium falciparum lines in vitro: Evidence for decreased sensitivity to dormancy and growth inhibition","volume":"56","author":"Teuscher","year":"2012","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1002\/biof.5520260201","article-title":"Antithrombotic and anticancer effects of garlic-derived sulfur compounds: A review","volume":"26","author":"Ariga","year":"2006","journal-title":"Biofactors"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"346","DOI":"10.1016\/0091-7435(90)90034-H","article-title":"Garlic: A review of its relationship to malignant disease","volume":"19","author":"Dausch","year":"1990","journal-title":"Prev. Med."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"57","DOI":"10.2131\/jts.9.57","article-title":"[Acute toxicity test of garlic extract]","volume":"9","author":"Nakagawa","year":"1984","journal-title":"J. Toxicol. Sci."},{"key":"ref_48","first-page":"181","article-title":"Differentiating and growth inhibitory effects of diallyl disulfide on cancer cells","volume":"11","author":"Lea","year":"1997","journal-title":"Int. J. Oncol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"700","DOI":"10.1128\/AAC.23.5.700","article-title":"Antifungal activity in human urine and serum after ingestion of garlic (Allium sativum)","volume":"23","author":"Caporaso","year":"1983","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1200","DOI":"10.1016\/S0891-5849(03)00144-8","article-title":"Comparative effects of mono-, di-, tri-, and tetrasulfides derived from plants of the allium family: Redox cycling in vitro and hemolytic activity and phase 2 enzyme induction in vivo","volume":"34","author":"Munday","year":"2003","journal-title":"Free Radic. Biol. Med."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"558","DOI":"10.1177\/1091581816655303","article-title":"Toxicological Evaluation of beta-Caryophyllene Oil: Subchronic Toxicity in Rats","volume":"35","author":"Schmitt","year":"2016","journal-title":"Int. J. Toxicol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"304","DOI":"10.1016\/j.ijpharm.2013.04.013","article-title":"Physicochemical characterization and pharmacokinetics evaluation of beta-caryophyllene\/beta-cyclodextrin inclusion complex","volume":"450","author":"Liu","year":"2013","journal-title":"Int. J. Pharm."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1680","DOI":"10.1016\/j.foodchem.2011.02.036","article-title":"Physicochemical and release characterisation of garlic oil-beta-cyclodextrin inclusion complexes","volume":"127","author":"Wang","year":"2011","journal-title":"Food Chem."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"1218","DOI":"10.1126\/science.7355284","article-title":"Babesia bovis: Continuous cultivation in a microaerophilous stationary phase culture","volume":"207","author":"Levy","year":"1980","journal-title":"Science"}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/9\/6\/466\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:38:19Z","timestamp":1760175499000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/9\/6\/466"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,6,12]]},"references-count":54,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2020,6]]}},"alternative-id":["pathogens9060466"],"URL":"https:\/\/doi.org\/10.3390\/pathogens9060466","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,6,12]]}}}