{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T09:39:13Z","timestamp":1790501953118,"version":"4.1.0"},"reference-count":39,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2021,6,1]],"date-time":"2021-06-01T00:00:00Z","timestamp":1622505600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000199","name":"U.S. Department of Agriculture","doi-asserted-by":"publisher","award":["2018-67015-28295"],"award-info":[{"award-number":["2018-67015-28295"]}],"id":[{"id":"10.13039\/100000199","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vaccines"],"abstract":"<jats:p>Enterobactin (Ent) is a highly conserved and important siderophore for the growth of many Gram-negative bacterial pathogens. Therefore, targeting Ent for developing innovative intervention strategies has attracted substantial research interest in recent years. Recently, we developed a novel Ent conjugate vaccine that has been demonstrated to be effective for controlling Gram-negative pathogens using both in vitro and in vivosystems. In particular, active immunization of chickens with the Ent conjugate vaccine elicited strong immune responses and significantly reduced intestinal colonization of Campylobacter jejuni, the leading foodborne bacterial pathogen. Given that hyperimmune egg yolk immunoglobulin Y (IgY) has been increasingly recognized as a promising and practical non-antibiotic approach for passive immune protection against pathogens in livestock, in this study, we assessed the efficacy of oral administration of broiler chickens with the anti-Ent hyperimmune egg yolk powder to control C. jejuni colonization in the intestine. However, supplementation of feed with 2% (w\/w) of anti-Ent egg yolk powder failed to reduce C. jejuni colonization when compared to the control group. Consistent with this finding, the ELISA titers of the specific IgY in cecum, ileum, duodenum, gizzard, and serum contents were similar between the two groups throughout the trial. Chicken intestinal microbiota also did not change in response to the egg yolk powder treatment. Subsequently, to examine ex vivo stability of the egg yolk IgY, the chicken gizzard and duodenum contents from two independent sources were spiked with the egg yolk antibodies, incubated at 42 \u00b0C for different lengths of time, and subjected to ELISA analysis. The specific IgY titers were dramatically decreased in gizzard contents (up to 2048-fold) but were not changed in duodenum contents. Collectively, oral administration of broiler chickens with the anti-Ent egg yolk powder failed to confer protection against intestinal colonization of C. jejuni, which was due to instability of the IgY in gizzard contents as demonstrated by both in vivo and ex vivo evidence.<\/jats:p>","DOI":"10.3390\/vaccines9060569","type":"journal-article","created":{"date-parts":[[2021,5,31]],"date-time":"2021-05-31T23:33:33Z","timestamp":1622504013000},"page":"569","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Passive Immunization of Chickens with Anti-Enterobactin Egg Yolk Powder for Campylobacter Control"],"prefix":"10.3390","volume":"9","author":[{"given":"Huiwen","family":"Wang","sequence":"first","affiliation":[{"name":"Department of Animal Science, The University of Tennessee, Knoxville, TN 37996, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2795-6988","authenticated-orcid":false,"given":"Ximin","family":"Zeng","sequence":"additional","affiliation":[{"name":"Department of Animal Science, The University of Tennessee, Knoxville, TN 37996, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6560-154X","authenticated-orcid":false,"given":"Liu","family":"Cao","sequence":"additional","affiliation":[{"name":"Department of Animal Science, The University of Tennessee, Knoxville, TN 37996, USA"},{"name":"Department of Civil and Environmental Engineering, The University of Tennessee, Knoxville, TN 37996, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7155-6474","authenticated-orcid":false,"given":"Qiang","family":"He","sequence":"additional","affiliation":[{"name":"Department of Civil and Environmental Engineering, The University of Tennessee, Knoxville, TN 37996, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0377-1030","authenticated-orcid":false,"given":"Jun","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Animal Science, The University of Tennessee, Knoxville, TN 37996, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,6,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1146\/annurev-food-022811-101137","article-title":"Egg yolk antibodies for passive immunity","volume":"3","author":"Mine","year":"2012","journal-title":"Annu. Rev. Food Sci. Technol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"860","DOI":"10.1016\/j.biotechadv.2011.07.003","article-title":"Application of chicken egg yolk immunoglobulins in the control of terrestrial and aquatic animal diseases: A review","volume":"29","author":"Xu","year":"2011","journal-title":"Biotechnol. Adv."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1017\/S1466252315000195","article-title":"Passive immunization with hyperimmune egg-yolk IgY as prophylaxis and therapy for poultry diseases\u2014A review","volume":"16","author":"Gadde","year":"2015","journal-title":"Anim. Health Res. Rev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"701","DOI":"10.1086\/509936","article-title":"The health burden of Campylobacter infection and the impact of antimicrobial resistance: Playing chicken","volume":"44","year":"2007","journal-title":"Clin. Infect. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1016\/j.trsl.2020.04.009","article-title":"New and alternative strategies for the prevention, control, and treatment of antibiotic-resistant Campylobacter","volume":"223","author":"Dai","year":"2020","journal-title":"Transl. Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"755","DOI":"10.1089\/fpd.2008.0247","article-title":"Novel approaches for Campylobacter control in poultry","volume":"6","author":"Lin","year":"2009","journal-title":"Foodborne Pathog. Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1079\/WPS20020016","article-title":"Factors affecting intestinal colonisation of poultry by Campylobacter and role of microflora in control","volume":"58","author":"Mead","year":"2002","journal-title":"Worlds Poult. Sci. J."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1017\/S1466252317000135","article-title":"Antibiotic resistance trends and mechanisms in the foodborne pathogen, Campylobacter","volume":"18","author":"Tang","year":"2017","journal-title":"Anim. Health Res. Rev."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e00319","DOI":"10.1128\/AEM.00358-19","article-title":"Enterobactin-specific antibodies induced by a novel enterobactin conjugate vaccine","volume":"85","author":"Wang","year":"2019","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_10","unstructured":"Sheppard, S.K. (2014). Siderophore-mediated iron acquisition for Campylobacter infection. Campylobacter Ecology and Evolution, Caister Academic."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1128\/MMBR.00012-07","article-title":"Siderophore-based iron acquisition and pathogen control","volume":"71","author":"Miethke","year":"2007","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"7764","DOI":"10.1016\/j.vaccine.2020.10.040","article-title":"Enterobactin-specific antibodies inhibit in vitro growth of different gram-negative bacterial pathogens","volume":"38","author":"Wang","year":"2020","journal-title":"Vaccine"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Cui, Y., Guo, F., Guo, J., Cao, X., Wang, H., Yang, B., Zhou, H., Su, X., Zeng, X., and Lin, J. (2020). Immunization of chickens with the enterobactin conjugate vaccine reduced Campylobacter jejuni colonization in the intestine. Vaccines, 8.","DOI":"10.3390\/vaccines8040747"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"779","DOI":"10.3389\/fimmu.2021.629480","article-title":"Evaluation of the immunogenic response of a novel enterobactin conjugate vaccine in chickens for the production of enterobactin-specific egg yolk antibodies","volume":"12","author":"Zeng","year":"2021","journal-title":"Front. Immunol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"3075","DOI":"10.1021\/acs.molpharmaceut.0c00492","article-title":"t-Butanol enables dual functionality of mannitol: A cryoprotectant in frozen systems and bulking agent in freeze-dried formulations","volume":"17","author":"Sonje","year":"2020","journal-title":"Mol. Pharm."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Diraviyam, T., Zhao, B., Wang, Y., Schade, R., Michael, A., and Zhang, X. (2014). Effect of chicken egg yolk antibodies (IgY) against diarrhea in domesticated animals: A systematic review and meta-analysis. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0097716"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Wang, Z., Li, J., Li, J., Li, Y., Wang, L., Wang, Q., Fang, L., Ding, X., Huang, P., and Yin, J. (2019). Protective effect of chicken egg yolk immunoglobulins (IgY) against enterotoxigenic Escherichia coli K88 adhesion in weaned piglets. BMC Vet. Res., 15.","DOI":"10.1186\/s12917-019-1958-x"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1111\/j.1439-0450.2004.00739.x","article-title":"Effect of orally administered egg yolk antibodies on Salmonella enteritidis contamination of hen\u2032s eggs","volume":"51","author":"Methner","year":"2004","journal-title":"J. Vet. Med. B"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4516","DOI":"10.1073\/pnas.1000080107","article-title":"Global patterns of 16S rRNA diversity at a depth of millions of sequences per sample","volume":"108","author":"Caporaso","year":"2011","journal-title":"Proc. Natl. Acad Sci. USA"},{"key":"ref_20","unstructured":"(2020, December 10). Illumina 16S Metagenomic Sequencing Library Preparation. Available online: https:\/\/support.illumina.com\/documents\/documentation\/chemistry_documentation\/16s\/16s-metagenomic-library-prep-guide-15044223-b.pdf."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"852","DOI":"10.1038\/s41587-019-0209-9","article-title":"Reproducible, interactive, scalable and extensible microbiome data science using QIIME 2","volume":"37","author":"Bolyen","year":"2019","journal-title":"Nat. Biotechnol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1016\/j.jbiotec.2017.06.1198","article-title":"25 years of serving the community with ribosomal RNA gene reference databases and tools","volume":"261","author":"Yilmaz","year":"2017","journal-title":"J. Biotechnol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"581","DOI":"10.1038\/nmeth.3869","article-title":"DADA2: High-resolution sample inference from Illumina amplicon data","volume":"13","author":"Callahan","year":"2016","journal-title":"Nat. Methods"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"McMurdie, P.J., and Holmes, S. (2013). phyloseq: An R package for reproducible interactive analysis and graphics of microbiome census data. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0061217"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"306","DOI":"10.3382\/japr.2014-00937","article-title":"Function of the digestive system","volume":"23","author":"Svihus","year":"2014","journal-title":"J. Appl. Poult. Res."},{"key":"ref_26","first-page":"108","article-title":"Activities of amylase, trypsin and chymotrypsin of pancreas and small intestinal contents in the red jungle fowl and broiler breed","volume":"10","author":"Kadhim","year":"2011","journal-title":"Afr. J. Biotechnol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1077","DOI":"10.1271\/bbb.57.1077","article-title":"Oral passive immunization effect of anti-human rotavirus IgY and its behavior against proteolytic enzymes","volume":"57","author":"Hatta","year":"1993","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"4638","DOI":"10.1016\/S0021-9258(18)93672-0","article-title":"Purification and characterization of chicken pepsinogen and chicken pepsin","volume":"244","author":"Bohak","year":"1969","journal-title":"J. Biol. Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1051\/rnd:19990404","article-title":"Comparison of pig and chicken pepsins for protein hydrolysis","volume":"39","author":"Gomez","year":"1999","journal-title":"Reprod. Nutr. Dev."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1016\/j.jim.2004.11.017","article-title":"Microencapsulation for the gastric passage and controlled intestinal release of immunoglobulin Y","volume":"296","author":"Mine","year":"2005","journal-title":"J. Immunol. Methods"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2911","DOI":"10.1021\/jf062900q","article-title":"Chitosan\u2212alginate microcapsules for oral delivery of egg yolk immunoglobulin (IgY)","volume":"55","author":"Li","year":"2007","journal-title":"J. Agric. Food Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"106349","DOI":"10.1016\/j.foodhyd.2020.106349","article-title":"Formulation of alginate\/carrageenan microgels to encapsulate, protect and release immunoglobulins: Egg Yolk IgY","volume":"112","author":"Gu","year":"2021","journal-title":"Food Hydrocoll."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"3785","DOI":"10.1016\/j.vaccine.2019.05.082","article-title":"Evaluation of in ovo vaccination of DNA vaccines for Campylobacter control in broiler chickens","volume":"37","author":"Liu","year":"2019","journal-title":"Vaccine"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2322","DOI":"10.3382\/ps\/pey081","article-title":"In ovo applications in poultry: A review","volume":"97","author":"Peebles","year":"2018","journal-title":"Poult. Sci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"60","DOI":"10.3389\/fnut.2018.00060","article-title":"The well-developed mucosal immune systems of birds and mammals allow for similar approaches of mucosal vaccination in both types of animals","volume":"5","author":"Nochi","year":"2018","journal-title":"Front. Nutr."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1073\/pnas.232686799","article-title":"Identification of pathogen-specific and conserved genes expressed in vivo by an avian pathogenic Escherichia coli strain","volume":"100","author":"Dozois","year":"2003","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"352","DOI":"10.1080\/03079457.2018.1475724","article-title":"Re-thinking the chicken\u2013Campylobacter jejuni interaction: A review","volume":"47","author":"Awad","year":"2018","journal-title":"Avian Pathol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"582","DOI":"10.1128\/MMBR.00015-13","article-title":"Salmonella pathogenicity and host adaptation in chicken-associated serovars","volume":"77","author":"Foley","year":"2013","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"ref_39","unstructured":"Kiyono, H., and Pascual, D.W. (2020). Current and new approaches for mucosal vaccine delivery. Mucosal Vaccines, Elsevier. [2nd ed.]."}],"container-title":["Vaccines"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-393X\/9\/6\/569\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:09:42Z","timestamp":1760162982000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-393X\/9\/6\/569"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,6,1]]},"references-count":39,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2021,6]]}},"alternative-id":["vaccines9060569"],"URL":"https:\/\/doi.org\/10.3390\/vaccines9060569","relation":{},"ISSN":["2076-393X"],"issn-type":[{"value":"2076-393X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,6,1]]}}}