{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,8]],"date-time":"2026-01-08T16:56:52Z","timestamp":1767891412413,"version":"3.49.0"},"reference-count":54,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2023,12,20]],"date-time":"2023-12-20T00:00:00Z","timestamp":1703030400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Indonesia Endowment Funds for Education (LPDP)","award":["PD1012023071000358"],"award-info":[{"award-number":["PD1012023071000358"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>This study investigates the potential role of Cold-pressed Valencia Terpeneless citrus oil (CO), as a natural antimicrobial, in controlling causative agents of pullorum disease and fowl typhoid in floor materials for poultry farming, specifically wooden chips. The study addresses the issues that have arisen as a result of the reduction in antibiotic use in poultry farming, which has resulted in the re-emergence of bacterial diseases including salmonellosis. CO efficiently inhibits the growth of pathogens including various serovars of Salmonella enterica (SE), including SE serovar Gallinarum (S. Gallinarum) and SE serovar Pullorum (S. Pullorum), in a dose-dependent manner. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of CO showed potential for controlling diverse S. Gallinarum and S. Pullorum isolates. Growth inhibition assays demonstrated that 0.4% (v\/w) CO eliminated S. Pullorum and S. Gallinarum from 24 h onwards, also impacting poultry gut microbiota and probiotic strains. Floor material simulation, specifically wooden chips treated with 0.4% CO, confirmed CO\u2019s effectiveness in preventing S. Gallinarum and S. Pullorum growth on poultry house floors. This study also investigated the effect of CO on the expression of virulence genes in S. Gallinarum and S. Pullorum. Specifically, the study revealed that the application of CO resulted in a downregulation trend in virulence genes, including spiA, invA, spaN, sitC, and sifA, in both S. Pullorum and S. Gallinarum, implying that CO may alter the pathogenicity of these bacterial pathogens. Overall, this study reveals that CO has the potential to be used as a natural antimicrobial in the prevention and management of Salmonella-related infections in chicken production, offering a viable alternative to control these re-emerging diseases.<\/jats:p>","DOI":"10.3390\/ani14010023","type":"journal-article","created":{"date-parts":[[2023,12,20]],"date-time":"2023-12-20T11:24:33Z","timestamp":1703071473000},"page":"23","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Unveiling the Potential Ways to Apply Citrus Oil to Control Causative Agents of Pullorum Disease and Fowl Typhoid in Floor Materials"],"prefix":"10.3390","volume":"14","author":[{"given":"Dita","family":"Julianingsih","sequence":"first","affiliation":[{"name":"Department of Animal and Avian Sciences, University of Maryland, College Park, MD 20742, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3850-4813","authenticated-orcid":false,"given":"Chuan-Wei","family":"Tung","sequence":"additional","affiliation":[{"name":"Department of Animal and Avian Sciences, University of Maryland, College Park, MD 20742, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kanchan","family":"Thapa","sequence":"additional","affiliation":[{"name":"Department of Animal and Avian Sciences, University of Maryland, College Park, MD 20742, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Debabrata","family":"Biswas","sequence":"additional","affiliation":[{"name":"Department of Animal and Avian Sciences, University of Maryland, College Park, MD 20742, USA"},{"name":"Biological Sciences Program, University of Maryland, College Park, MD 20742, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,12,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"100310","DOI":"10.1016\/j.japr.2022.100310","article-title":"Poultry industry paradigms: Connecting the dots","volume":"32","author":"Castro","year":"2023","journal-title":"J. Appl. Poult. Res."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Grzybowska-Brzezi\u0144ska, M., Banach, J.K., and Grzywi\u0144ska-R\u0105pca, M. (2023). Shaping Poultry Meat Quality Attributes in the Context of Consumer Expectations and Preferences-A Case Study of Poland. Foods, 12.","DOI":"10.3390\/foods12142694"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1002\/app5.377","article-title":"A policy and research agenda for Asia\u2019s poultry industry","volume":"10","author":"Mahanty","year":"2023","journal-title":"Asia Pac. Policy Stud."},{"key":"ref_4","unstructured":"USDA (United States Department of Agriculture) (2023, December 09). Animal Raising Claims Labeling Guidelines, Available online: https:\/\/www.fsis.usda.gov\/sites\/default\/files\/media_file\/2021-09\/Animal-Raising-Claims-labeling-and-Non-GMO-slides-2021-09-01.pdf."},{"key":"ref_5","unstructured":"Kirchhelle, C. (2023, November 06). Pyrrhic Progress: The History of Antibiotics in Anglo-American Food Production, Available online: https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/32101389."},{"key":"ref_6","unstructured":"WHO (World Health Organization) (2023, November 29). Antibiotic Resistance. Available online: https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/antibiotic-resistance#:~:text=Antibiotic%20resistance%20is%20accelerated%20by,poor%20infection%20prevention%20and%20control."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1016\/j.bsheal.2020.09.004","article-title":"Use of antimicrobials in food animals and impact of transmission of antimicrobial resistance on humans","volume":"3","author":"Ma","year":"2021","journal-title":"Biosaf. Health"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1016\/S2542-5196(17)30141-9","article-title":"Restricting the use of antibiotics in food-producing animals and its associations with antibiotic resistance in food-producing animals and human beings: A systematic review and meta-analysis","volume":"1","author":"Tang","year":"2017","journal-title":"Lancet Planet. Health"},{"key":"ref_9","unstructured":"Le, T. (2023, November 06). Antibiotic-Free Poultry Production: All You Need to Know. Alltech Blog. Available online: https:\/\/www.alltech.com\/blog\/antibiotic-free-poultry-production-all-you-need-know#:~:text=What%20are%20common%20challenges%20of,a%20decrease%20in%20growth%20performance."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"101864","DOI":"10.1016\/j.psj.2022.101864","article-title":"Effects of therapeutic levels of dietary antibiotics on the cecal microbiome composition of broiler chickens","volume":"101","author":"Kairmi","year":"2022","journal-title":"Poult. Sci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"279","DOI":"10.3382\/japr\/pfx066","article-title":"Vaccines and vaccination against fowl typhoid and pullorum disease: An overview and approaches in developing countries","volume":"27","author":"Revolledo","year":"2018","journal-title":"J. Appl. Poult. Res."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Kipper, D., Mascitti, A.K., De Carli, S., Carneiro, A.M., Streck, A.F., Fonseca, A.S.K., Ikuta, N., and Lunge, V.R. (2022). Emergence, Dissemination and Antimicrobial Resistance of the Main Poultry-Associated Salmonella Serovars in Brazil. Vet. Sci., 9.","DOI":"10.3390\/vetsci9080405"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Xiong, D., Song, L., Pan, Z., and Jiao, X. (2018). Identification and Discrimination of Salmonella enterica Serovar Gallinarum Biovars Pullorum and Gallinarum Based on a One-Step Multiplex PCR Assay. Front. Microbiol., 9.","DOI":"10.3389\/fmicb.2018.01718"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Li, C., Xu, Z., Chen, W., Zhou, C., Wang, C., Wang, M., Liang, J., and Wei, P. (2022). The Use of Star Anise-Cinnamon Essential Oil as an Alternative Antibiotic in Prevention of Salmonella Infections in Yellow Chickens. Antibiotics, 11.","DOI":"10.3390\/antibiotics11111579"},{"key":"ref_15","unstructured":"CFSPH (The Center for Food Security and Public Health) (2022, December 13). Fowl Typhoid and Pullorum Disease. Available online: https:\/\/www.cfsph.iastate.edu\/Factsheets\/pdfs\/fowl_typhoid.pdf."},{"key":"ref_16","unstructured":"Yeakel, S.D. (2019). Merck Manual (Veterinary Manual), Available online: https:\/\/www.merckvetmanual.com\/poultry\/salmonelloses\/pullorum-disease-in-poultry?gclid=Cj0KCQiAj4ecBhD3ARIsAM4Q_jGK343rYs0z7G7nymH9XZWn46_2jWoIXR8SyOiKKFZwkxLje2GmjYAaAs-cEALw_wcBandgclsrc=aw.ds."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"405","DOI":"10.20506\/rst.19.2.1222","article-title":"Fowl typhoid and pullorum disease","volume":"19","author":"Shivaprasad","year":"2000","journal-title":"Rev. Sci. Tech."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"107671","DOI":"10.1016\/j.pharmthera.2020.107671","article-title":"Efficacy and mechanism of actions of natural antimicrobial drugs","volume":"216","author":"Ye","year":"2020","journal-title":"Pharmacol. Ther."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1021\/bk-2018-1287.ch001","article-title":"Natural and Bio-based Antimicrobials: A Review. In Natural and Bio-Based Antimicrobials for Food Applications","volume":"1287","author":"Fan","year":"2018","journal-title":"ACS Symp. Ser."},{"key":"ref_20","first-page":"M264","article-title":"Orange Essential Oils Antimicrobial Activities against Salmonella spp.","volume":"73","author":"Crandall","year":"2008","journal-title":"J. Food Sci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1020","DOI":"10.1111\/j.1365-2672.2012.05270.x","article-title":"Antimicrobial effect and mode of action of terpeneless cold-pressed Valencia orange essential oil on methicillin-resistant Streptococcus aureus","volume":"112","author":"Muthaiyan","year":"2012","journal-title":"J. Appl. Microbiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1089\/fpd.2011.0946","article-title":"Enhanced inhibition of Listeria monocytogenes by a combination of cold-pressed terpeneless Valencia orange oil and antibiotics","volume":"9","author":"Milillo","year":"2012","journal-title":"Foodborne Pathog. Dis."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"695","DOI":"10.1089\/fpd.2008.0124","article-title":"Antimicrobial activity of commercial citrus-based natural extracts against Escherichia coli O157:H7 isolates and mutant strains","volume":"5","author":"Nannapaneni","year":"2008","journal-title":"Foodborne Pathog. Dis."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1128\/am.19.1.27-31.1970","article-title":"Antimicrobial action of some citrus fruit oils on selected food-borne bacteria","volume":"19","author":"Dabbah","year":"1970","journal-title":"Appl. Microbiol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"688","DOI":"10.1099\/jmm.0.000286","article-title":"Major components of orange oil inhibit Streptococcus aureus growth and biofilm formation and alter its virulence factors","volume":"65","author":"Federman","year":"2016","journal-title":"J. Med. Microbiol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"653","DOI":"10.1080\/14786419.2016.1219860","article-title":"Essential oil components of orange peels and antimicrobial activity","volume":"31","author":"Geraci","year":"2017","journal-title":"Nat. Prod. Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"114535","DOI":"10.1016\/j.lwt.2023.114535","article-title":"Integrated transcriptome reveals that d-limonene inhibits Candida tropicalis by disrupting metabolism","volume":"176","author":"Xiong","year":"2023","journal-title":"LWT"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e12918","DOI":"10.1111\/jfs.12918","article-title":"Antimicrobial activity and mechanism of limonene against Streptococcus aureus","volume":"41","author":"Han","year":"2021","journal-title":"J. Food Saf."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Guo, F., Chen, Q., Liang, Q., Zhang, M., Chen, W., Chen, H., Yun, Y., Zhong, Q., and Chen, W. (2021). Antimicrobial Activity and Proposed Action Mechanism of Linalool Against Pseudomonas fluorescens. Front. Microbiol., 12.","DOI":"10.3389\/fmicb.2021.562094"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"870","DOI":"10.1002\/jsfa.3897","article-title":"Microbial inhibitory and radical scavenging activities of cold-pressed terpeneless Valencia orange (Citrus sinensis) oil in different dispersing agents","volume":"90","author":"Chalova","year":"2010","journal-title":"J. Sci. Food Agric."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Birhanu, B.T., Lee, E.B., Lee, S.J., and Park, S.C. (2021). Targeting Salmonella Typhimurium Invasion and Intracellular Survival Using Pyrogallol. Front. Microbiol., 12.","DOI":"10.3389\/fmicb.2021.631426"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Alvarado-Martinez, Z., Bravo, P., Kennedy, N.F., Krishna, M., Hussain, S., Young, A.C., and Biswas, D. (2020). Antimicrobial and antivirulence impacts of phenolics on Salmonella enterica serovar Typhimurium. Antibiotics, 9.","DOI":"10.3390\/antibiotics9100668"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1637\/11927-071318-ResNote.1","article-title":"Virulence gene distribution of Salmonella Pullorum isolates recovered from chickens in China (1953\u20132015)","volume":"62","author":"Zhang","year":"2018","journal-title":"Avian Dis."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1637\/7417.1","article-title":"Virulence genotyping of Salmonella spp. with multiplex PCR","volume":"50","author":"Skyberg","year":"2006","journal-title":"Avian Dis."},{"key":"ref_35","first-page":"325","article-title":"The antibacterial effects of terpene alcohols on Streptococcus aureus and their mode of action","volume":"237","author":"Inoue","year":"2004","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_36","unstructured":"Chen, Y., Ni, J., and Li, H. (2023, November 14). Avian Leukosis Virus Subgroup J Infection Influencing Composition of Gut Microbiota within Chicken. Huizhou University, Southern Medical University. Available online: https:\/\/assets.researchsquare.com\/files\/rs-16010\/v2\/335be006-a5a1-4148-9191-d8575c2da516.pdf?c=1631841451."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"3551","DOI":"10.1016\/j.watres.2004.05.021","article-title":"Presence of Enterococcus faecalis in broiler litter and wild bird feces for bacterial source tracking","volume":"38","author":"Kuntz","year":"2004","journal-title":"Water Res."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Priyodip, P., and Balaji, S. (2019). An in vitro chicken gut model for the assessment of phytase producing bacteria. 3 Biotech, 9.","DOI":"10.1007\/s13205-019-1825-2"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"450","DOI":"10.1016\/j.aninu.2023.06.001","article-title":"Probiotic Escherichia coli Nissle 1917 protect chicks from damage caused by Salmonella enterica serovar Enteritidis colonization","volume":"14","author":"Wu","year":"2023","journal-title":"Anim. Nutr."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"7800","DOI":"10.1073\/pnas.93.15.7800","article-title":"Identification of a pathogenicity island required for Salmonella survival in host cells","volume":"93","author":"Ochman","year":"1996","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1132","DOI":"10.1016\/j.meegid.2010.07.015","article-title":"Association between phylogeny, virulence potential, and serovars of Salmonella enterica","volume":"10","author":"Litrup","year":"2010","journal-title":"Infect. Genet. Evol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1186\/s13620-017-0095-8","article-title":"Molecular characterization of antimicrobial resistant non-typhoidal Salmonella from poultry industries in Korea","volume":"70","author":"Kim","year":"2017","journal-title":"Ir. Vet. J."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Lou, L., Zhang, P., Piao, R., and Wang, Y. (2019). Salmonella Pathogenicity Island 1 (SPI-1) and Its Complex Regulatory Network. Front. Cell. Infect. Microbiol., 9.","DOI":"10.3389\/fcimb.2019.00270"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"4338","DOI":"10.1128\/jb.174.13.4338-4349.1992","article-title":"Molecular and functional characterization of the Salmonella invasion gene invA: Homology of invA to members of a new protein family","volume":"174","author":"Ginocchio","year":"1992","journal-title":"J. Bacteriol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"562","DOI":"10.5455\/vetworld.2011.562-564","article-title":"Inv A gene-specific PCR for detection of Salmonella from broilers","volume":"4","author":"Shanmugasamy","year":"2011","journal-title":"Vet. World"},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Mohammed, B.T. (2024). Identification and bioinformatic analysis of invA gene of Salmonella in free-range chicken. Braz. J. Biol., 84.","DOI":"10.1590\/1519-6984.263363"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"e36","DOI":"10.1590\/s1678-9946201961036","article-title":"Detection of virulence genes in Salmonella Heidelberg isolated from chicken carcasses","volume":"61","author":"Webber","year":"2019","journal-title":"Rev. Do Inst. Med. Trop. S\u00e3o Paulo"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1016\/j.micpath.2004.04.006","article-title":"Characterization of Salmonella pathogenicity island 1 type III secretion-dependent haemolytic activity in Salmonella enterica serovar Typhimurium","volume":"37","author":"Miki","year":"2004","journal-title":"Microb. Pathog."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"567","DOI":"10.1038\/nature05272","article-title":"Protein delivery into eukaryotic cells by type III secretion machines","volume":"444","author":"Galan","year":"2006","journal-title":"Nature"},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Myeni, S.K., Wang, L., and Zhou, D. (2013). SipB-SipC complex is essential for translocon formation. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0060499"},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Beshiru, A., Igbinosa, I.H., and Igbinosa, E.O. (2019). Prevalence of Antimicrobial Resistance and Virulence Gene Elements of Salmonella Serovars From Ready-to-Eat (RTE) Shrimps. Front. Microbiol., 10.","DOI":"10.3389\/fmicb.2019.01613"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"33151","DOI":"10.1074\/jbc.M109.034975","article-title":"Interaction between the sifA virulence factor and its host target SKIP is essential for Salmonella pathogenesis","volume":"284","author":"Diacovich","year":"2009","journal-title":"J. Biol. Chem."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1080\/19490976.2018.1506668","article-title":"Caspase-3 cleavage of Salmonella type III secreted effector protein sifA is required for localization of functional domains and bacterial dissemination","volume":"10","author":"Patel","year":"2019","journal-title":"Gut Microbes"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"12979","DOI":"10.1038\/srep12979","article-title":"The Salmonella effector protein sifA plays a dual role in virulence","volume":"5","author":"Zhao","year":"2015","journal-title":"Sci. Rep."}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/14\/1\/23\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:42:15Z","timestamp":1760132535000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/14\/1\/23"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,12,20]]},"references-count":54,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2024,1]]}},"alternative-id":["ani14010023"],"URL":"https:\/\/doi.org\/10.3390\/ani14010023","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,12,20]]}}}