{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,16]],"date-time":"2026-06-16T06:49:31Z","timestamp":1781592571831,"version":"3.54.5"},"reference-count":74,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2025,3,3]],"date-time":"2025-03-03T00:00:00Z","timestamp":1740960000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2025,3,3]],"date-time":"2025-03-03T00:00:00Z","timestamp":1740960000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100007102","name":"Zagazig University","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100007102","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["BMC Vet Res"],"abstract":"<jats:title>Abstract<\/jats:title>\n          <jats:sec>\n            <jats:title>Background<\/jats:title>\n            <jats:p>\n              <jats:italic>Salmonella enterica<\/jats:italic> serovar Pullorum, the causative agent of pullorum disease, is one cause of the economic losses in the global poultry industry. Vaccination and antibiotics are still the most effective methods of controlling Salmonella, even though the vaccine contains the causative agent, and the antibiotic therapy has limited efficacy. We provide a novel immunostimulator and antibiotic substitute to protect against and avoid <jats:italic>Salmonella pullorum<\/jats:italic> (SP) infection.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Methods<\/jats:title>\n            <jats:p>\n              <jats:italic>Nigella sativa<\/jats:italic>-purified oil (NS) and monophosphoryl lipid A (MPLA) were formulated as nanoliposomal compounds (NS-MPLA). Their protective and immunomodulatory efficacies were experimentally tested orally in broiler chicks against challenge with virulent pandemic drug-resistant SP. Four chick groups were utilized: control; NS-MPLA-supplemented; SP-challenged; and SP-challenged, then NS-MPLA-treated. Clinical signs, organ gross pathology, colony-forming counts, and tissue histopathological alterations were investigated. The relative fold-changes in the expression of <jats:italic>IL-1\u03b2, IL-4, IL-17<\/jats:italic>, <jats:italic>IL-22, TLR-4, INF-\u03b3, IgA,<\/jats:italic> and <jats:italic>MUC2<\/jats:italic> genes were evaluated.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>The SP-challenged chicks showed notable symptoms and extensive pathological lesions in their internal organs. The bacteria colonized the challenged chicks' livers and continued to shed in their feces for 5\u20136\u00a0days. A minor amount of immune cell tissue trafficking was noted. The NS-MPLA-treated chicks displayed opposing patterns after being challenged with SP. They exhibited mild clinical signs with modest gross pathology in the internal organs. After 3\u20134\u00a0days, the liver and the fecal droppings were cleared of SP. Significant heterophilic aggregation, lymphocytic infiltration, and lymphoid follicle enlargement were observed. Additionally, chicks challenged with SP and then NS-MPLA-treated showed a 5- to tenfold increase in immune-related cytokines, immunoglobulin A, and mucosal relative gene expression folds compared to the SP-challenged non-NS-MPLA-treated, which showed a sharp decline in <jats:italic>IL-4<\/jats:italic> and <jats:italic>IL-22<\/jats:italic> and a minor rise in the rest of the tested gene relative expressions. Chicks given NS-MPLA supplementation showed a significant upregulation of these genes compared to the control group.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusion<\/jats:title>\n            <jats:p>In this first report on poultry, it is possible to draw the conclusion that NS-MPLA supplementation in SP-infected chicks boosts immunity and provides protection. It promoted bacterial clearance and tissue repair and stimulated the expression of genes linked to immunity and the mucosal surface. These findings suggest the potential application of NS-MPLA in salmonella control programs as an antibiotic substitute or in immunization strategies.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1186\/s12917-025-04473-w","type":"journal-article","created":{"date-parts":[[2025,3,3]],"date-time":"2025-03-03T02:06:33Z","timestamp":1740967593000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Nigella sativa monophosphoryl lipid A nanoliposome: a promising antibiotic alternative and immunomodulator to control virulent pandemic drug-resistant\u00a0Salmonella pullorum infection in broiler chicks"],"prefix":"10.1186","volume":"21","author":[{"given":"Adel Attia M.","family":"Ahmad","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elsayed Alsaied Masoud","family":"Hussien","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alaa A. A. M.","family":"Elian","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohamed","family":"Abdelmoneim","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A.","family":"Ali","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ahmed E.","family":"Abdelhamid","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gamal A.","family":"Elmowalid","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,3,3]]},"reference":[{"key":"4473_CR1","doi-asserted-by":"publisher","first-page":"165","DOI":"10.1016\/j.vetmic.2018.11.032","volume":"228","author":"R Guo","year":"2019","unstructured":"Guo R, Li Z, Zhou X, Huang C, Hu Y, Geng S, et al. Induction of arthritis in chickens by infection with novel virulent Salmonella Pullorum strains. Vet Microbial. 2019;228:165\u201372.","journal-title":"Vet Microbial"},{"issue":"3","key":"4473_CR2","doi-asserted-by":"publisher","first-page":"279","DOI":"10.3382\/japr\/pfx066","volume":"27","author":"L Revolledo","year":"2018","unstructured":"Revolledo L. Vaccines and vaccination against fowl typhoid and pullorum disease: An overview and approaches in developing countries. J Appl Poultry Res. 2018;27(3):279\u201391.","journal-title":"J Appl Poultry Res"},{"issue":"3","key":"4473_CR3","doi-asserted-by":"publisher","first-page":"100894","DOI":"10.1016\/j.psj.2020.12.007","volume":"100","author":"F Sun","year":"2021","unstructured":"Sun F, Li X, Wang Y, Wang F, Ge H, Pan Z, Xu Y, Wang Y, Jiao X, Chen X. Epidemic patterns of antimicrobial resistance of Salmonella enteric serovar Gallinarum biovar Pullorum isolates in China during the past half-century. Poult Sci. 2021;100(3):100894.","journal-title":"Poult Sci"},{"issue":"1","key":"4473_CR4","doi-asserted-by":"publisher","first-page":"23","DOI":"10.3390\/antibiotics13010023","volume":"13","author":"M Farhat","year":"2024","unstructured":"Farhat M, Khayi S, Berrada J, Mouahid M, Ameur N, El-Adawy H, Fellahi S. Salmonella enterica Serovar Gallinarum Biovars Pullorum and Gallinarum in Poultry: Review of Pathogenesis, Antibiotic Resistance, Diagnosis and Control in the Genomic Era. Antibiotics. 2024;13(1):23.","journal-title":"Antibiotics"},{"key":"4473_CR5","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1016\/j.vetimm.2016.03.015","volume":"173","author":"R Guo","year":"2016","unstructured":"Guo R, Geng S, Jiao H, Pan Z, Chen X, Jiao X. Evaluation of protective efficacy of a novel inactivated Salmonella Pullorum ghost vaccine against virulent challenge in chickens. Vet Immunopathol. 2016;173:27\u201333.","journal-title":"Vet Immunopathol"},{"key":"4473_CR6","doi-asserted-by":"publisher","first-page":"1","DOI":"10.2903\/j.efsa.2004.114","volume":"114","author":"EFSA","year":"2004","unstructured":"EFSA. Opinion of the Scientific Panel on Biological Hazards on a request from the Commission related to the use of vaccines for the control of Salmonella in poultry. The EFSA J. 2004;114:1\u201374. https:\/\/doi.org\/10.2903\/j.efsa.2004.114.","journal-title":"The EFSA Journal"},{"issue":"5","key":"4473_CR7","doi-asserted-by":"publisher","first-page":"282","DOI":"10.3389\/fmicb.2014.00282","volume":"17","author":"MS Diarra","year":"2014","unstructured":"Diarra MS, Malouin F. Antibiotics in Canadian poultry productions and anticipated alternatives. Front Microbiol. 2014;17(5):282. https:\/\/doi.org\/10.3389\/fmicb.2014.00282. PMID: 24987390; PMCID: PMC4060556","journal-title":"Front Microbiol"},{"issue":"5","key":"4473_CR8","doi-asserted-by":"publisher","first-page":"247","DOI":"10.54846\/jshap\/1016","volume":"25","author":"LL Schulz","year":"2017","unstructured":"Schulz LL, Rademacher CJ. Food and Drug Administration Guidance 209 and 213 and Veterinary Feed Directive regulations regarding antibiotic use in livestock: a survey of preparation and anticipated impacts in the swine industry. J Swine Health Prod. 2017;25(5):247\u201355.","journal-title":"J Swine Health Prod"},{"key":"4473_CR9","unstructured":"Asheg AA, Abdullatif M Karmous, Ben Naser KM, Abuzaid L. Duration of Feeding Black seed (Nigella sativa) to Broiler Chicks and Its Effect on Immune Response, Cholesterol and Gut Microflora.Res J Pharmaceut Biol and Chem Sci. 2016;7(1):183\u2013187."},{"key":"4473_CR10","doi-asserted-by":"publisher","unstructured":"Talebi A, Maham M, Asri-Rezaei S, Pournaghi P, Khorrami MS, Derakhshan A. Effects of Nigella sativa on Performance, Blood Profiles, and Antibody Titer against Newcastle Disease in Broilers. Evid Based Complement Alternat Med. 2021;14(2021):2070375. https:\/\/doi.org\/10.1155\/2021\/2070375. PMID34234833PMCID: PMC8216824.","DOI":"10.1155\/2021\/2070375"},{"key":"4473_CR11","doi-asserted-by":"publisher","unstructured":"Soliman E S, Hamad R T, Ahmed A. Prophylactic and immune modulatory influences of\u00a0Nigella sativa\u00a0Linn. In broilers exposed to biological challenge. Vet World. 2017; 10(12):1447\u20131455. https:\/\/doi.org\/10.14202\/vetworld.2017.1447-1455. Epub 2017 Dec 12. PMID: 29391685; PMCID: PMC5771169.","DOI":"10.14202\/vetworld.2017.1447-1455"},{"key":"4473_CR12","doi-asserted-by":"publisher","unstructured":"Manjunatha V, Nixon J E, Mathis G F, Lumpkins B S, G\u00fczel-Seydim Z B, Seydim A C, Greene A K, Jiang X. Nigella sativa as an antibiotic alternative to promote growth and enhance health of broilers challenged with Eimeria maxima and Clostridium perfringens. Poult Sci. 2023 Aug; 102(8):102831. https:\/\/doi.org\/10.1016\/j.psj.2023.102831. Epub 2023 Jun 3. Erratum in: Poult Sci. 2023 Oct; 102(10):103015. https:\/\/doi.org\/10.1016\/j.psj.2023.103015. PMID: 37356299; PMCID: PMC10404756.","DOI":"10.1016\/j.psj.2023.102831 10.1016\/j.psj.2023.103015"},{"key":"4473_CR13","unstructured":"Abu Dieyeh, A.Z. and Darwish, A.M. Effect of Feeding Powdered Black Cumin Seeds (Nigella sativa L .) on Growth Performance of 4\u20138 Week-Old Broilers. J Animal Vet Adv. 2008;7(3):292\u2013296."},{"issue":"2\u20133","key":"4473_CR14","doi-asserted-by":"publisher","first-page":"251","DOI":"10.1016\/S0378-8741(02)00324-0","volume":"84","author":"HS El-Abhar","year":"2003","unstructured":"El-Abhar HS, Abdallah DM, Saleh S. Gastroprotective activity of Nigella sativa oil and its constituent, thymoquinone, against gastric mucosal injury induced by ischaemia\/reperfusion in rats. J Ethno Pharmacol. 2003;84(2\u20133):251\u20138.","journal-title":"J Ethno pharmacol"},{"issue":"1","key":"4473_CR15","first-page":"652","volume":"6","author":"I Intsar Kadhim","year":"2018","unstructured":"Intsar Kadhim I, Al-Zubaidi TM, Haider AH. Influence of dietary supplementation of Nigella sativa on experimental coccidiosis in broiler chickens. Journal of Entomology and Zoology Studies. 2018;6(1):652\u20136.","journal-title":"Journal of Entomology and Zoology Studies"},{"issue":"1","key":"4473_CR16","doi-asserted-by":"publisher","first-page":"6568","DOI":"10.1038\/s41598-023-33416-4","volume":"13","author":"Nu Lail","year":"2023","unstructured":"Lail Nu, Sattar A, Omer MO, et al. Biosynthesis and characterization of zinc oxide nanoparticles using Nigella sativa against coccidiosis in commercial poultry. Sci Rep. 2023;13(1):6568. https:\/\/doi.org\/10.1038\/s41598-023-33416-4.","journal-title":"Sci Rep"},{"issue":"3","key":"4473_CR17","doi-asserted-by":"publisher","first-page":"404","DOI":"10.1016\/j.jsps.2016.09.011","volume":"25","author":"ZT Rushmi","year":"2017","unstructured":"Rushmi ZT, Akter N, Mow RJ, Afroz M, Kazi M, de Matas M, Rahman M, Shariare MH. The impact of formulation attributes and process parameters on black seed oil loaded liposomes and their performance in animal models of analgesia. Saudi Pharm J. 2017;25(3):404\u201312.","journal-title":"Saudi Pharm J"},{"key":"4473_CR18","doi-asserted-by":"publisher","unstructured":"Ali T, Hussain F, Naeem M, Khan A, Al-Harrasi A. Nanotechnology Approach for Exploring the Enhanced Bioactivities and Biochemical Characterization of Freshly Prepared\u00a0Nigella sativa\u00a0L. Nano suspensions and Their Phytochemical Profile. Front Bioeng Biotechnol. 2022 May 17; 10:888177. https:\/\/doi.org\/10.3389\/fbioe.2022.888177.","DOI":"10.3389\/fbioe.2022.888177"},{"issue":"19","key":"4473_CR19","doi-asserted-by":"publisher","first-page":"6744","DOI":"10.3390\/molecules27196744","volume":"27","author":"MH Shariare","year":"2022","unstructured":"Shariare MH, Khan MA, Al-Masum A, Khan JH, Uddin J, Kazi M. Development of Stable Liposomal Drug Delivery System of Thymoquinone and Its In vitro Anticancer Studies Using Breast Cancer and Cervical Cancer Cell Lines. Molecules. 2022;27(19):6744. https:\/\/doi.org\/10.3390\/molecules27196744. PMID: 36235288; PMCID: PMC9571792.","journal-title":"Molecules"},{"key":"4473_CR20","doi-asserted-by":"publisher","unstructured":"Ghiaee Shamloo, A.; Yadegari, M.H.; Jaafari, M.R.; Zarrinfar, H. Inhibitory Effect Evaluation of\u00a0Nigella sativa\u00a0Oil Loaded to Liposomal Nanocarriers against\u00a0Candida parapsilosis\u00a0Isolates.\u00a0Preprints.\u00a02023;2023061953. https:\/\/doi.org\/10.20944\/preprints202306.1953.v1.","DOI":"10.20944\/preprints202306.1953.v1"},{"key":"4473_CR21","doi-asserted-by":"publisher","unstructured":"Childers N K, Miller K L, Tong G, Llarena JC, Green way T, Ulrich JT, Michalek SM. Adjuvant activity of monophosphoryl lipid A for nasal and oral immunization with soluble or liposome-associated antigen. Infect Immun. 2000;68(10):5509\u201316. https:\/\/doi.org\/10.1128\/IAI.68.10.5509-5516.2000. PMID: 10992447; PMCID: PMC101499.","DOI":"10.1128\/IAI.68.10.5509-5516.2000"},{"issue":"2","key":"4473_CR22","doi-asserted-by":"publisher","first-page":"103","DOI":"10.18176\/jiaci.0349","volume":"29","author":"JM Zubeldia","year":"2019","unstructured":"Zubeldia JM, Ferrer M, D\u00e1vila I, Justicia JL. Adjuvants in Allergen-Specific Immunotherapy: Modulating and Enhancing the Immune Response. J Investig Allergol Clin Immunol. 2019;29(2):103\u201311. https:\/\/doi.org\/10.18176\/jiaci.0349. (Epub 2018 Nov 12 PMID: 30418155).","journal-title":"J Investig Allergol Clin Immunol"},{"key":"4473_CR23","doi-asserted-by":"publisher","unstructured":"Heath MD, Mohsen MO, de Kam PJ, Carreno Velazquez TL, Hewings SJ, Kramer MF, K\u00fcndig TM, Bachmann MF, Skinner MA. Shaping Modern Vaccines: Adjuvant Systems Using Micro Crystalline Tyrosine (MCT\u00ae). Front Immunol. 2020;24(11):594911. https:\/\/doi.org\/10.3389\/fimmu.2020.594911. PMID: 33324411; PMCID: PMC7721672.","DOI":"10.3389\/fimmu.2020.594911"},{"key":"4473_CR24","doi-asserted-by":"publisher","unstructured":"Lin YJ, Zimmermann J, Sch\u00fclke S. Novel adjuvants in allergen-specific immunotherapy: where do we stand? Front Immunol. 2024;23(15):1348305. https:\/\/doi.org\/10.3389\/fimmu.2024.1348305. PMID: 38464539; PMCID: PMC10920236.","DOI":"10.3389\/fimmu.2024.1348305"},{"key":"4473_CR25","doi-asserted-by":"publisher","unstructured":"Wang YQ, Bazin-Lee H, Evans JT, Casella CR, Mitchell TC. MPL Adjuvant Contains Competitive Antagonists of Human TLR4. Front Immunol. 2020;16(11):577823. https:\/\/doi.org\/10.3389\/fimmu.2020.577823. PMID: 33178204; PMCID: PMC7596181.","DOI":"10.3389\/fimmu.2020.577823"},{"issue":"5\u20136","key":"4473_CR26","doi-asserted-by":"publisher","first-page":"368","DOI":"10.1016\/S0264-410X(02)00617-5","volume":"21","author":"K Johansen","year":"2003","unstructured":"Johansen K, Schroder U, Svensson L. Immunogenicity and protective efficacy of a formalin-inactivated rotavirus vaccine combined with lipid adjuvants. Vaccine. 2003;21(5\u20136):368\u201375.","journal-title":"Vaccine"},{"issue":"7","key":"4473_CR27","doi-asserted-by":"publisher","first-page":"1129","DOI":"10.1517\/14712598.4.7.1129","volume":"4","author":"JR Baldridge","year":"2004","unstructured":"Baldridge JR, McGowan P, Evans JT, Cluff C, Mossman S, Johnson D, Persing D. Taking a Toll on human disease: Toll-like receptor 4 agonists as vaccine adjuvants and mono therapeutic agents. Expert Opin Biol Ther. 2004;4(7):1129\u201338.","journal-title":"Expert Opin Biol Ther"},{"key":"4473_CR28","doi-asserted-by":"publisher","unstructured":"Kang X, Yang Y, Meng C, Wang X, Liu B, Geng S, Jiao X, Pan Z. Safety and protective efficacy of Salmonella Pullorum spiC and rfaH deletion rough mutant as a live attenuated DIVA vaccine candidate. Poult Sci. 2022 Mar; 101(3):101655. https:\/\/doi.org\/10.1016\/j.psj.2021.101655. Epub 2021 Dec 8. PMID: 34991038; PMCID: PMC8743217.","DOI":"10.1016\/j.psj.2021.101655"},{"issue":"11","key":"4473_CR29","doi-asserted-by":"publisher","first-page":"5316","DOI":"10.1016\/j.psj.2020.07.017","volume":"99","author":"C Chen","year":"2020","unstructured":"Chen C, Li J, Zhang H, Xie Y, Xiong L, Liu H, Fang Wang F. Effects of a probiotic on the growth performance, intestinal flora, and immune function of chicks infected with Salmonella Pullorum. Poult Sci. 2020;99(11):5316\u201323.","journal-title":"Poult Sci"},{"issue":"90","key":"4473_CR30","doi-asserted-by":"publisher","first-page":"21","DOI":"10.1016\/j.colsurfb.2011.09.042","volume":"1","author":"W Fan","year":"2012","unstructured":"Fan W, Yan W, Xu Z, Ni H. Formation mechanism of monodisperse, low molecular weight chitosan nanoparticles by ionic gelation technique. Colloids Surf B Biointerfaces. 2012;1(90):21\u20137. https:\/\/doi.org\/10.1016\/j.colsurfb.2011.09.042. (Epub 2011 Oct 2 PMID: 22014934).","journal-title":"Colloids Surf B Biointerfaces"},{"key":"4473_CR31","unstructured":"Ramadan Mansour E, El-Saber MM, Abdelhamid AE, Ahmed A. El-Sayed, Effect of nano chitosan encapsulated spermine on growth, productivity and bioactive compounds of chili pepper (Capsicum annuum L.) under salinity stress. Egyptian J Chem. 2022;65(8):187\u2013198."},{"key":"4473_CR32","doi-asserted-by":"publisher","first-page":"717","DOI":"10.2307\/1592106","volume":"38","author":"M Nakamura","year":"1994","unstructured":"Nakamura M, Nagamine N, Takahashi T, Suzuki S, Sato S. Evaluation of the efficacy of a bacterin against Salmonella enteritidis infection and the effect of stress after vaccination. Avian Dis. 1994;38:717\u201324.","journal-title":"Avian Dis"},{"key":"4473_CR33","unstructured":"ISO. Standards catalogue 07.100.30. Food Microbiology. 2011. http:\/\/www.iso.org\/iso\/products\/standards\/catalogu."},{"key":"4473_CR34","doi-asserted-by":"publisher","unstructured":"Xiong D, Song L, Geng S, Tao J, An S, Pan Z, Jiao X. One-Step PCR Detection of Salmonella Pullorum\/Gallinarum Using a Novel Target: The Flagellar Biosynthesis Gene flhB. Front Microbiol. 2016;22(7):1863. https:\/\/doi.org\/10.3389\/fmicb.2016.01863. PMID: 27920764; PMCID: PMC5118417.","DOI":"10.3389\/fmicb.2016.01863"},{"key":"4473_CR35","doi-asserted-by":"publisher","unstructured":"Weinstein MP, Lewis JS 2nd. The Clinical and Laboratory Standards Institute Subcommittee on Antimicrobial Susceptibility Testing: Background, Organization, Functions, and Processes. J Clin Microbiol. 2020;58(3):e01864-e1919. https:\/\/doi.org\/10.1128\/JCM.01864-19. PMID: 31915289; PMCID: PMC7041576.","DOI":"10.1128\/JCM.01864-19"},{"issue":"4","key":"4473_CR36","doi-asserted-by":"publisher","first-page":"46","DOI":"10.5409\/wjcp.v2.i4.46","volume":"2","author":"XZ Huang","year":"2013","unstructured":"Huang XZ, Zhu LB, Li ZR, Lin J. Bacterial colonization and intestinal mucosal barrier development. World J Clin Pediatr. 2013;2(4):46\u201353.","journal-title":"World J Clin Pediatr"},{"key":"4473_CR37","doi-asserted-by":"crossref","unstructured":"Suvarna SK, MBBS, BSC, FRCP, FRCPath. Cardiac Pathology - A Guide To Current Practice 2013. In: Suvarna SK, editor. 3rd ed. London: Springer; 2013. p. 1\u201317.","DOI":"10.1007\/978-1-4471-2407-8_1"},{"key":"4473_CR38","doi-asserted-by":"publisher","unstructured":"Ghareeb K, Awad WA, Soodoi C, Sasgary S, Strasser A, B\u00f6hm J. Effects of feed contaminant deoxynivalenol on plasma cytokines and mRNA expression of immune genes in the intestine of broiler chickens. PLoSOne. 2013;8(8):e71492. https:\/\/doi.org\/10.1371\/journal.pone.0071492. Erratum. In: PLoSOne. 2014;9(6):e100051. PMID: 23977054; PMCID: PMC3748120.","DOI":"10.1371\/journal.pone.0071492"},{"key":"4473_CR39","doi-asserted-by":"publisher","unstructured":"Liu H, Zhang M, Han H, Yuan J, Li Z. Comparison of the expression of cytokine genes in the bursal tissues of the chickens following challenge with infectious bursal disease viruses of varying virulence. Virol J. 2010;8(7):364. https:\/\/doi.org\/10.1186\/1743-422X-7-364.","DOI":"10.1186\/1743-422X-7-364"},{"issue":"2","key":"4473_CR40","doi-asserted-by":"publisher","first-page":"172","DOI":"10.3382\/ps\/peu064","volume":"94","author":"Q Zhang","year":"2015","unstructured":"Zhang Q, Eicher SD, Applegate TJ. Development of intestinal mucin-2, IgA, and polymeric Ig receptor expressions in broiler chickens and Pekin ducks. Poult Sci. 2015;94(2):172\u201380.","journal-title":"Poult Sci"},{"issue":"4","key":"4473_CR41","doi-asserted-by":"publisher","first-page":"402","DOI":"10.1006\/meth.2001.1262","volume":"25","author":"KJ Livak","year":"2001","unstructured":"Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2-\u0394\u0394CT method. Methods. 2001;25(4):402\u20138.","journal-title":"Methods"},{"key":"4473_CR42","doi-asserted-by":"publisher","unstructured":"Elmowalid GA, Ahmad AAM, El-Hamid MIA, Ibrahim D, Wahdan A, El Oksh ASA, Yonis AE, Elkady MA, Ismail TA, Alkhedaide AQ, Elnahriry SS. Nigella sativa Extract Potentially Inhibited Methicillin Resistant Staphylococcus aureus Induced Infection in Rabbits: Potential Immunomodulatory and Growth Promoting Properties. Animals (Basel). 2022;12(19):2635. https:\/\/doi.org\/10.3390\/ani12192635. PMID: 36230379; PMCID: PMC9559630.","DOI":"10.3390\/ani12192635"},{"key":"4473_CR43","doi-asserted-by":"publisher","first-page":"103","DOI":"10.1186\/s12917-023-03632-1","volume":"19","author":"AAM Ahmad","year":"2023","unstructured":"Ahmad AAM, Elmowalid GA, Abdelhamid AE, et al. Nigella sativa-chitosan nanoparticles: Novel intestinal mucosal immunomodulator controls and protects against Salmonella enterica serovar Enteritidis infection in broilers. BMC Vet Res. 2023;19:103.","journal-title":"BMC Vet Res"},{"key":"4473_CR44","doi-asserted-by":"crossref","unstructured":"Elmowalid G A, Ammar A M, Ahmad A A M. Nigella sativa seed extract: 1. Enhancement of sheep macrophage immune functions in vitro. Res Vet Sci. 2013; 95 (2):437\u201343.","DOI":"10.1016\/j.rvsc.2013.02.015"},{"issue":"9","key":"4473_CR45","doi-asserted-by":"publisher","first-page":"e1006512","DOI":"10.1371\/journal.ppat.1006512","volume":"13","author":"SA Ragland","year":"2017","unstructured":"Ragland SA, Criss AK. From bacterial killing to immune modulation: Recent insights into the functions of lysozyme. PLoS Pathog. 2017;13(9):e1006512.","journal-title":"PLoS Pathog"},{"key":"4473_CR46","doi-asserted-by":"publisher","unstructured":"El-Sharkawy H, Tahoun A, El-Gohary AEA, El-Abasy M, El-Khayat F, Gillespie T, Kitade Y, Hafez HM, Neubauer H, El-Adawy H. Epidemiological, molecular characterization and antibiotic resistance of Salmonella enterica serovars isolated from chicken farms in Egypt. Gut Pathog. 2017Feb;10(9):8. https:\/\/doi.org\/10.1186\/s13099-017-0157-1. PMID: 28203289; PMCID: PMC5301364.","DOI":"10.1186\/s13099-017-0157-1"},{"issue":"12","key":"4473_CR47","doi-asserted-by":"publisher","first-page":"1435","DOI":"10.3390\/antibiotics10121435","volume":"10","author":"T Ramatla","year":"2021","unstructured":"Ramatla T, Tawana M, Onyiche TE, Lekota KE, Thekisoe O. Prevalence of antibiotic resistance in salmonella serotypes concurrently isolated from the environment, animals, and humans in South Africa: a systematic review and meta-analysis. Antibiotics (Basel). 2021;10(12):1435.","journal-title":"Antibiotics (Basel)"},{"key":"4473_CR48","doi-asserted-by":"publisher","first-page":"3545","DOI":"10.1007\/s13197-020-04360-2","volume":"57","author":"T Subramani","year":"2020","unstructured":"Subramani T, Ganapathyswamy H. An overview of liposomal nano-encapsulation techniques and its applications in food and nutraceutical. J Food Sci Technol. 2020;57:3545\u201355. https:\/\/doi.org\/10.1007\/s13197-020-04360-2.","journal-title":"J Food Sci Technol"},{"key":"4473_CR49","doi-asserted-by":"publisher","first-page":"1421","DOI":"10.1007\/s00289-023-04768-8","volume":"81","author":"AE Abdelhamid","year":"2024","unstructured":"Abdelhamid AE, Ahmed EH, Awad HM, et al. Synthesis and cytotoxic activities of selenium nanoparticles incorporated nano-chitosan. Polym Bull. 2024;81:1421\u201337. https:\/\/doi.org\/10.1007\/s00289-023-04768-8.","journal-title":"Polym Bull"},{"issue":"1\u20133","key":"4473_CR50","doi-asserted-by":"publisher","first-page":"81","DOI":"10.1016\/s0169-409x(01)00171-5","volume":"51","author":"L Illum","year":"2001","unstructured":"Illum L, Jabbal-Gill I, Hinchcliffe M, Fisher AN, Davis SS. Chitosan as a novel nasal delivery system for vaccines. Adv Drug Deliv Rev. 2001;51(1\u20133):81\u201396. https:\/\/doi.org\/10.1016\/s0169-409x(01)00171-5. (PMID: 11516781).","journal-title":"Adv Drug Deliv Rev"},{"key":"4473_CR51","doi-asserted-by":"publisher","first-page":"761","DOI":"10.2147\/IJN.S238445","volume":"15","author":"S Renu","year":"2020","unstructured":"Renu S, Markazi AD, Dhakal S, Lakshmanappa YS, Shanmugasundaram R, Selvaraj RK, et al. Oral deliverable mucoadhesive Chitosan-Salmonella subunit nanovaccine for layer chickens. Int J Nanomedicine. 2020;15:761\u201377. https:\/\/doi.org\/10.2147\/IJN.S238445.","journal-title":"Int J Nanomedicine"},{"key":"4473_CR52","volume-title":"Novel strategies for improved mucosal immunity in chickens: Application to control salmonellosis","author":"WI Muir","year":"1996","unstructured":"Muir WI. Novel strategies for improved mucosal immunity in chickens: Application to control salmonellosis. PhD diss.: University of Sydney; 1996."},{"key":"4473_CR53","doi-asserted-by":"crossref","unstructured":"Dar MA, Urwat U, Ahmad SM, Ahmad R, Kashoo ZA, Dar TA, Bhat SA, Mumtaz PT, Shabir N, Shah RA, Heidari M. Gene expression and antibody response in chicken against Salmonella Typhimurium challenge. Poult Sci. 2019;98(5):2008\u20132013.","DOI":"10.3382\/ps\/pey560"},{"key":"4473_CR54","first-page":"14","volume":"84","author":"PA Chacana","year":"2003","unstructured":"Chacana PA, Terzolo HR. New light on old views of avian Pullorosis: avian Typhosis. Rev Inst Med Trop. 2003;84:14\u201320.","journal-title":"Rev Inst Med Trop"},{"issue":"10","key":"4473_CR55","doi-asserted-by":"publisher","first-page":"e2487","DOI":"10.1371\/journal.pntd.0002487","volume":"7","author":"BN Parsons","year":"2013","unstructured":"Parsons BN, Humphrey S, Salisbury AM, Mikoleit J, Hinton JC, Gordon MA, Wigley P. Invasive Non- Typhoidal Salmonella Typhimurium ST313 Are Not Host- Restricted and Have an Invasive Phenotype in Experimentally. PLoS Negl Trop Dis. 2013;7(10):e2487.","journal-title":"PLoS Negl Trop Dis"},{"key":"4473_CR56","doi-asserted-by":"publisher","first-page":"83","DOI":"10.1007\/s00580-011-1371-8","volume":"22","author":"SN Shah","year":"2013","unstructured":"Shah SN, Kamil SA, Darzi MM, et al. Haematological and some biochemical changes in experimental fowl typhoid infection in broiler chickens. Comp Clin Pathol. 2013;22:83\u201391. https:\/\/doi.org\/10.1007\/s00580-011-1371-8.","journal-title":"Comp Clin Pathol"},{"issue":"5","key":"4473_CR57","doi-asserted-by":"publisher","first-page":"2986","DOI":"10.1128\/IAI.73.5.2986-2990.2005","volume":"73","author":"P Wigley","year":"2005","unstructured":"Wigley P, Hulme SD, Powers C, Beal RK, Berchieri A Jr, Smith A, Barrow P. Infection of the reproductive tract and eggs with Salmonella enterica serovar pullorum in the chicken is associated with suppression of cellular immunity at sexual maturity. Infect Immun. 2005;73(5):2986\u201390.","journal-title":"Infect Immun"},{"key":"4473_CR58","doi-asserted-by":"crossref","unstructured":"Al-Mufarrej, S.I. Immune-responsiveness and performance of broiler chickens fed black cumin (Nigella Sativa L.) powder. Journal of the Saudi Soc Agric Sci. 2014;13(1):75-80.","DOI":"10.1016\/j.jssas.2013.01.006"},{"issue":"2","key":"4473_CR59","first-page":"76","volume":"2","author":"T Azeem","year":"2014","unstructured":"Azeem T, Rehmanb ZU, Umar S, Asifa M, Arifc M, Rahmanc A. Effect of Nigella sativa on poultry health and production: A review. Sci Lett. 2014;2(2):76\u201382.","journal-title":"Sci Lett"},{"issue":"7","key":"4473_CR60","doi-asserted-by":"publisher","first-page":"305","DOI":"10.1016\/s1471-4922(03)00122-3","volume":"19","author":"DM Moncada","year":"2003","unstructured":"Moncada DM, Kammanadiminti SJ, Chadee K. Mucin and Toll-like receptors in host defense against intestinal parasites. Trends Parasitol. 2003Jul;19(7):305\u201311. https:\/\/doi.org\/10.1016\/s1471-4922(03)00122-3. (PMID: 12855381).","journal-title":"Trends Parasitol"},{"issue":"1","key":"4473_CR61","doi-asserted-by":"publisher","first-page":"45","DOI":"10.1038\/nrc1251","volume":"4","author":"MA Hollingsworth","year":"2004","unstructured":"Hollingsworth MA, Swanson BJ. Mucins in cancer: Protection and control of the cell surface. Nat Rev Cancer. 2004;4(1):45\u201360.","journal-title":"Nat Rev Cancer"},{"key":"4473_CR62","doi-asserted-by":"publisher","first-page":"189","DOI":"10.1016\/j.bbcan.2006.01.002","volume":"1765","author":"M Andrianifahanana","year":"2006","unstructured":"Andrianifahanana M, Moniaux N, Batra SK. Regulation of mucin expression: Mechanistic aspects and implications for cancer and inflammatory diseases. Biochim Bio phys Acta-Rev Cancer. 2006;1765:189\u2013222.","journal-title":"Biochim Bio phys Acta-Rev Cancer"},{"issue":"6","key":"4473_CR63","doi-asserted-by":"publisher","first-page":"411","DOI":"10.1038\/nri2316","volume":"8","author":"D Artis","year":"2008","unstructured":"Artis D. Epithelial-cell recognition of commensal bacteria and maintenance of immune homeostasis in the gut. Nat Rev Immunol. 2008;8(6):411\u201320. https:\/\/doi.org\/10.1038\/nri2316. (PMID: 18469830).","journal-title":"Nat Rev Immunol"},{"key":"4473_CR64","doi-asserted-by":"crossref","unstructured":"Sadarman Febrina D, Yendraliza Shirothul HM, Nurfitriani AR, Barkah NN, Sholikin MM, Yunilas Qomariyah N, Anuraga Jayanegara A, Solfaine R, Irawan A. Effect of dietary black cumin seed (Nigella sativa) on performance, immune status, and serum metabolites of small ruminants: A meta-analysis. Small Ruminant Res. 2021;204:106521.","DOI":"10.1016\/j.smallrumres.2021.106521"},{"issue":"13","key":"4473_CR65","doi-asserted-by":"publisher","DOI":"10.3389\/fphys.2022.920777","volume":"18","author":"K Acevedo-Villanueva","year":"2022","unstructured":"Acevedo-Villanueva K, Akerele G, Al-Hakeem W, Adams D, Gourapura R, Selvaraj R. Immunization of Broiler Chickens with a Killed Chitosan Nanoparticle Salmonella Vaccine Decreases Salmonella Enterica Serovar Enteritidis Load. Front Physiol. 2022;18(13):920777. https:\/\/doi.org\/10.3389\/fphys.2022.920777.","journal-title":"Front Physiol"},{"key":"4473_CR66","doi-asserted-by":"publisher","unstructured":"Tiruppur Venkatachallam SK, Pattekhan H, Divakar S, Kadimi US. Chemical composition of Nigella sativa L. seed extracts obtained by supercritical carbon dioxide. J Food Sci Technol. 2010; 47(6):598\u2013605. https:\/\/doi.org\/10.1007\/s13197-010-0109-y. Epub 2010 Nov 1. PMID: 23572692; PMCID: PMC3551137.","DOI":"10.1007\/s13197-010-0109-y"},{"key":"4473_CR67","doi-asserted-by":"publisher","first-page":"113349","DOI":"10.1016\/j.biopha.2022.113349","volume":"153","author":"K Ciesielska-Figlon","year":"2022","unstructured":"Ciesielska-Figlon K, Daca A, Kokotkiewicz A, \u0141uczkiewicz M, Zabiega\u0142a B, Witkowski JM, Lisowska KA. The influence of Nigella sativa essential oil on proliferation, activation, and apoptosis of human T lymphocytes in vitro. Biomed Pharmacother. 2022;153:113349. https:\/\/doi.org\/10.1016\/j.biopha.2022.113349. Epub 2022 Jun 29 PMID: 35779419.","journal-title":"Biomed Pharmacother"},{"key":"4473_CR68","doi-asserted-by":"publisher","unstructured":"Ouattar H, Zouirech O, Kara M, Assouguem A, Almutairi SM, Al-Hemaid FM, Rasheed RA, Ullah R, Abbasi AM, Aouane M, Mikou K. In Vitro Study of the Phytochemical Composition and Antioxidant, Immunostimulant, and Hemolytic Activities of Nigella sativa (Ranunculaceae) and Lepidium sativum Seeds. Molecules. 2022;27(18):5946. https:\/\/doi.org\/10.3390\/molecules27185946. PMID: 36144678; PMCID: PMC9505328.","DOI":"10.3390\/molecules27185946"},{"issue":"2","key":"4473_CR69","doi-asserted-by":"publisher","first-page":"268","DOI":"10.1016\/j.jep.2010.06.030","volume":"131","author":"AF Majdalawieh","year":"2010","unstructured":"Majdalawieh AF, Hmaidan R, Carr RI. Nigella sativa modulates splenocyte proliferation, Th1\/Th2 cytokine profile, macrophage function and NK anti-tumor activity. J Ethnopharmacol. 2010;131(2):268\u201375.","journal-title":"J Ethnopharmacol"},{"issue":"4","key":"4473_CR70","doi-asserted-by":"publisher","first-page":"579","DOI":"10.30954\/2277-940X.04.2021.3","volume":"11","author":"Y Singh","year":"2021","unstructured":"Singh Y, Prakashbhai PN, Saxena B, Ramakrishnan S. Immunomodulatory Effects of Nigella sativa Extract on Chicken Peripheral Blood Mononuclear Cells: In-vitro. J Anim Res. 2021;11(4):579\u201386.","journal-title":"J Anim Res"},{"issue":"3","key":"4473_CR71","doi-asserted-by":"publisher","first-page":"201","DOI":"10.1631\/jzus.B1000163","volume":"12","author":"MH Boskabady","year":"2011","unstructured":"Boskabady MH, Keyhanmanesh R, Khameneh S, Doostdar Y, Khakzad MR. Potential immunomodulation effect of the extract of Nigella sativa on ovalbumin sensitized guinea pigs. J Zhejiang UnivSci B. 2011;12(3):201\u20139.","journal-title":"J Zhejiang UnivSci B"},{"issue":"3","key":"4473_CR72","doi-asserted-by":"publisher","first-page":"401","DOI":"10.1016\/j.dci.2012.08.002","volume":"38","author":"WH Kim","year":"2012","unstructured":"Kim WH, Jeong J, Park AR, Yim D, Kim YH, Kim KD, Chang HH, Lillehoj HS, Lee BH, Min W. Chicken IL-17F: identification and comparative expression analysis in Eimeria-infected chickens. Dev Comp Immunol. 2012;38(3):401\u20139. https:\/\/doi.org\/10.1016\/j.dci.2012.08.002.","journal-title":"Dev Comp Immunol"},{"issue":"8","key":"4473_CR73","doi-asserted-by":"publisher","first-page":"857","DOI":"10.1038\/ni.1767","volume":"10","author":"T Duhen","year":"2009","unstructured":"Duhen T, Geiger R, Jarrossay D, Lanzavecchia A, Sallusto F. Production of interleukin 22 but not interleukin 17 by a subset of human skin-homing memory T cells. Nat Immunol. 2009;10(8):857\u201363.","journal-title":"Nat Immunol"},{"issue":"9","key":"4473_CR74","doi-asserted-by":"publisher","first-page":"2369","DOI":"10.1002\/eji.201040848","volume":"40","author":"S Trifari","year":"2010","unstructured":"Trifari S, Spits H. IL-22-producing CD4+ T cells: middle-men between the immune system and its environment. Eur J Immunol. 2010;40(9):2369\u201371.","journal-title":"Eur J Immunol"}],"container-title":["BMC Veterinary Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12917-025-04473-w.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s12917-025-04473-w\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12917-025-04473-w.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,3,3]],"date-time":"2025-03-03T13:55:31Z","timestamp":1741010131000},"score":1,"resource":{"primary":{"URL":"https:\/\/bmcvetres.biomedcentral.com\/articles\/10.1186\/s12917-025-04473-w"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,3]]},"references-count":74,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["4473"],"URL":"https:\/\/doi.org\/10.1186\/s12917-025-04473-w","relation":{},"ISSN":["1746-6148"],"issn-type":[{"value":"1746-6148","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,3,3]]},"assertion":[{"value":"21 July 2024","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"3 January 2025","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"3 March 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"This experiment was carried out in conformity with ethical procedures and policies guidelines approved by the Institutional Animal Care and Use Committee of Zagazig University, ZU-IACUC, under number ZU-IACUC\/2\/F\/148\/2024.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"The authors declare no competing interests.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"132"}}