{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,12]],"date-time":"2026-05-12T08:44:51Z","timestamp":1778575491180,"version":"3.51.4"},"reference-count":52,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,3,7]],"date-time":"2022-03-07T00:00:00Z","timestamp":1646611200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Physiol."],"abstract":"<jats:p>Two experiments were conducted to investigate the effects of induced moisture loss on embryonic development and the immune response following an inflammatory challenge immediately post-hatch. In Experiment I, fertile leghorn eggs (<jats:italic>n<\/jats:italic> = 100) and commercial broiler eggs (<jats:italic>n<\/jats:italic> = 300) were set at 37.5\u00b0C and moisture loss was induced in one-half of the Leghorn and broiler eggs by drilling two, 1.5 mm diameter holes. The Control eggs had 0 holes. At embryonic day (ED)18, layer and broiler eggs in the 2-holes treatment had a significant (<jats:italic>P<\/jats:italic> &amp;lt; 0.01) increase in moisture loss compared to the control treatment (10.1% vs. 8.2%). Similarly, at ED18, the broiler eggs with 2-holes had a significant increase (<jats:italic>P<\/jats:italic> &amp;lt; 0.01) in moisture loss compared with control eggs (9.9% vs. 8.4%). Thymocytes from both the leghorn (104%) and broiler (62%) embryos in the 2-holes treatment had significantly increased <jats:italic>in vitro<\/jats:italic> proliferation compared with the control embryos (<jats:italic>P<\/jats:italic> \u2264 0.05). At ED18, layer and broiler embryos in the 2-holes treatment had an approximate twofold increase in the splenic CD8<jats:sup>+<\/jats:sup>\/CD4<jats:sup>+<\/jats:sup> ratio (<jats:italic>P<\/jats:italic> \u2264 0.05) and CD4<jats:sup>+<\/jats:sup>CD25<jats:sup>+<\/jats:sup> cells percentage in both the thymus and spleen (<jats:italic>P<\/jats:italic> \u2264 0.05). At ED18, both layer and broiler embryos from the 2-holes treatment had a significant increase in splenic IL1-\u03b2, IL-6, IL-10, and TLR-4 mRNA transcription compared to the control group (<jats:italic>P<\/jats:italic> \u2264 0.05). Experiment II was repeated with 300 fertile broiler eggs. On the day of hatch, chicks were randomly distributed into one of four treatments in a 2 (0, 2 holes) \u00d7 2 (0, 500 \u03bcg lipopolysaccharide, LPS) factorial arrangement of treatments. Chicks in the LPS groups were injected intraperitoneally with 500 \u03bcg\/kg BW LPS. At 24 and 48 h post-hatch, chicks hatched from eggs with 2-holes and challenged with LPS had a significant increase (<jats:italic>P<\/jats:italic> \u2264 0.05) in thymocyte proliferation at 24 h (42%) and 48 h (37%) when compared with chicks hatched from the control (0-hole; 0 \u03bcg LPS) treatment. Chicks hatched from the 2-holes treatment and challenged with the LPS had an approximately twofold higher splenic CD8<jats:sup>+<\/jats:sup>\/CD4<jats:sup>+<\/jats:sup> ratio and 1.5 fold increase in CD4<jats:sup>+<\/jats:sup>CD25<jats:sup>+<\/jats:sup> percentage compared to control chicks (P \u2264 0.05). In chicks hatched from the 2-holes treatment, MUC2 mRNA transcription was comparable to control chicks at 24 and 48 h in response to the LPS challenge. Our data suggest that the 2-holes treatment reprograms gene transcription to facilitate cell survival via proliferation and differentiation during an LPS inflammatory challenge.<\/jats:p>","DOI":"10.3389\/fphys.2022.820349","type":"journal-article","created":{"date-parts":[[2022,3,7]],"date-time":"2022-03-07T08:48:35Z","timestamp":1646642915000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":3,"title":["Effects of Induced Moisture Loss in Chicken Embryos at Embryonic Day 18 and Post-hatch Immune Response During Salmonella enteritidis Lipopolysaccharide Challenge in Broilers"],"prefix":"10.3389","volume":"13","author":[{"given":"Jenna L.","family":"Gregorich","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael S.","family":"Lilburn","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Revathi","family":"Shanmugasundaram","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2022,3,7]]},"reference":[{"key":"B1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1086\/405244","article-title":"Ontogeny of volume regulations in embryonic extracellular fluids.","volume":"42","author":"Adolph","year":"1967","journal-title":"Q. Rev. Biol."},{"key":"B2","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1093\/icb\/20.2.373","article-title":"Water in the avian egg overall budget of incubation.","volume":"20","author":"Ar","year":"1980","journal-title":"Am. Zool."},{"key":"B3","author":"Barott","year":"1937","journal-title":"Effect of Temperature, Humidity, and Other Factors on Hatch of Hens\u2019 Eggs and on Energy Metabolism of Chick Embryos."},{"key":"B4","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1515\/bmc-2012-0001","article-title":"The role of hyperosmotic stress in inflammation and disease.","volume":"3","author":"Brocker","year":"2012","journal-title":"Biomol. Concepts"},{"key":"B5","doi-asserted-by":"publisher","first-page":"827","DOI":"10.1093\/ps\/79.6.827","article-title":"Effects of relative humidity during incubation on hatchability and body weight of broiler chicks from young breeder flocks.","volume":"79","author":"Bruzual","year":"2000","journal-title":"Poult. Sci."},{"key":"B6","doi-asserted-by":"publisher","first-page":"874","DOI":"10.3382\/ps.0740874","article-title":"Incubation relative humidity effects on allantoic fluid volume and hatchability.","volume":"74","author":"Buhr","year":"1995","journal-title":"Poult. Sci."},{"key":"B7","author":"Cartwright","year":"2000","journal-title":"Incubating and Hatching Eggs."},{"key":"B8","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1097\/00024382-199507000-00007","article-title":"Hypertonic\/hyperoncotic fluids reverse prostaglandin E2 (PGE2)-induced T-cell suppression.","volume":"4","author":"Coimbra","year":"1995","journal-title":"Shock"},{"key":"B9","author":"Davis","year":"1987","journal-title":"The Effects of Variation in Egg Water Loss on the Chick Embryo and its Fluid Compartments."},{"key":"B10","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1002\/jez.1402450205","article-title":"Embryonic osmoregulation: consequences of high and low water loss during incubation of the chicken egg.","volume":"245","author":"Davis","year":"1988","journal-title":"J. Exp. Zool."},{"key":"B11","doi-asserted-by":"publisher","first-page":"574","DOI":"10.1080\/00071660802357025","article-title":"Comparison of a modern broiler and layer strain during embryonic development and the hatching process.","volume":"49","author":"Everaert","year":"2008","journal-title":"Br. Poult. Sci."},{"key":"B12","doi-asserted-by":"crossref","first-page":"892","DOI":"10.1590\/S0100-879X2009005000024","article-title":"Treatment of hemorrhagic shock with hypertonic saline solution modulates the inflammatory response to live bacteria in lungs.","volume":"42","author":"Fernandes","year":"2009","journal-title":"Braz. J. Med. Biol. Res."},{"key":"B13","doi-asserted-by":"publisher","first-page":"532","DOI":"10.1038\/nri3233","article-title":"Ion channels and transporters in lymphocyte function and immunity.","volume":"12","author":"Feske","year":"2012","journal-title":"Nat. Rev. Immunol."},{"key":"B14","doi-asserted-by":"crossref","first-page":"972","DOI":"10.1038\/nature05836","article-title":"Gene-specific control of inflammation by TLR-induced chromatin modifications.","volume":"447","author":"Foster","year":"2007","journal-title":"Nature"},{"key":"B15","doi-asserted-by":"crossref","first-page":"940741","DOI":"10.1155\/2010\/940741","article-title":"The chick chorioallantoic membrane: a model of molecular, structural, and functional adaptation to transepithelial ion transport and barrier function during embryonic development.","volume":"2010","author":"Gabrielli","year":"2010","journal-title":"J. Biomed. Biotechnol."},{"key":"B16","doi-asserted-by":"crossref","first-page":"738","DOI":"10.4049\/jimmunol.1501729","article-title":"TRPV4-mediated detection of hyposmotic stress by skin keratinocytes activates developmental immunity.","volume":"196","author":"Galindo-Villegas","year":"2016","journal-title":"J. Immunol."},{"key":"B17","doi-asserted-by":"crossref","first-page":"10673","DOI":"10.1073\/pnas.0403139101","article-title":"NFAT5\/TonEBP mutant mice define osmotic stress as a critical feature of the lymphoid microenvironment.","volume":"101","author":"Go","year":"2004","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"B18","doi-asserted-by":"crossref","first-page":"C787","DOI":"10.1152\/ajpcell.1986.251.5.C787","article-title":"Embryonic chick allantois: functional isolation and development of sodium transport.","volume":"251","author":"Graves","year":"1986","journal-title":"Am. J. Physiol. Cell Physiol."},{"key":"B19","first-page":"7","article-title":"The value of understanding moisture loss in incubation.","volume":"31","author":"Green","year":"2017","journal-title":"Int. Hatchery Pract."},{"key":"B20","doi-asserted-by":"crossref","first-page":"1499","DOI":"10.3382\/ps.0701499","article-title":"Effects of air humidity during incubation and age after hatch on heat tolerance of neonatal male and female chicks.","volume":"70","author":"Hamdy","year":"1991","journal-title":"Poult. Sci."},{"key":"B21","doi-asserted-by":"crossref","first-page":"354","DOI":"10.1086\/physzool.52.3.30155756","article-title":"Osmoregulation by avian embryos: the allantois functions like a toad\u2019s bladder.","volume":"52","author":"Hoyt","year":"1979","journal-title":"Physiol. Zool."},{"key":"B22","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1016\/j.biochi.2004.07.009","article-title":"Osmotic stress, a proinflammatory signal in Caco-2 cells.","volume":"86","author":"Hubert","year":"2004","journal-title":"Biochimie"},{"key":"B23","doi-asserted-by":"publisher","first-page":"757","DOI":"10.1080\/00071669608417905","article-title":"Selection for egg shell strength in laying hens using shell membrane characteristics.","volume":"37","author":"Johansson","year":"1996","journal-title":"Br. Poult. Sci."},{"key":"B24","author":"Landauer","year":"1967","journal-title":"Hatchability of Chicken Eggs as Influenced by Environment and Heredity."},{"key":"B25","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1093\/intimm\/dxt065","article-title":"MiR-568 inhibits the activation and function of CD4+ T cells and Treg cells by targeting NFAT5.","volume":"26","author":"Li","year":"2014","journal-title":"Int. Immunol."},{"key":"B26","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2- \u0394\u0394CT method.","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"B27","first-page":"143","article-title":"A review of the effects of temperature, humidity, turning and gaseous environment in the incubator on the hatchability of the hen\u2019s egg","author":"Lundy","year":"1969","journal-title":"The Fertility and Hatchability of Hen\u2019s Egg"},{"key":"B28","doi-asserted-by":"crossref","first-page":"45266","DOI":"10.1074\/jbc.M314132200","article-title":"Hyperosmotic stress induces rapid focal adhesion kinase phosphorylation at tyrosines 397 and 577: role of Src family kinases and Rho family GTPases.","volume":"279","author":"Lunn","year":"2004","journal-title":"J. Biol. Chem."},{"key":"B29","doi-asserted-by":"crossref","first-page":"2587","DOI":"10.1016\/j.molimm.2010.06.011","article-title":"Gene expression reprogramming protects macrophage from septic-induced cell death.","volume":"47","author":"Melo","year":"2010","journal-title":"Mol. Immunol."},{"key":"B30","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1186\/1742-9994-10-23","article-title":"The effect of hydration state and energy balance on innate immunity of a desert reptile.","volume":"10","author":"Moeller","year":"2013","journal-title":"Front. Zool."},{"key":"B31","doi-asserted-by":"publisher","first-page":"1716","DOI":"10.3382\/ps.2010-00779","article-title":"Effect of eggshell temperature and a hole in the air cell on the perinatal development and physiology of layer hatchlings.","volume":"89","author":"Molenaar","year":"2010","journal-title":"Poult. Sci."},{"key":"B32","article-title":"22.1. Osmoregulation and osmotic balance","author":"Molnar","year":"2013","journal-title":"Concepts of Biology-1st Canadian Edition."},{"key":"B33","doi-asserted-by":"publisher","first-page":"420","DOI":"10.3389\/fvets.2019.00420","article-title":"Dietary antibiotic growth promoters down-regulate intestinal inflammatory cytokine expression in chickens challenged with LPS or Co-infected with eimeria maxima and clostridium perfringens.","volume":"6","author":"Oh","year":"2019","journal-title":"Front. Vet. Sci."},{"key":"B34","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1093\/icb\/20.2.329","article-title":"The physics of gas exchange across the avian eggshell.","volume":"20","author":"Paganelli","year":"1980","journal-title":"Am. Zool."},{"key":"B35","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1186\/s12950-019-0220-4","article-title":"Hypertonic solution-induced preconditioning reduces inflammation and mortality rate.","volume":"16","author":"Pimentel","year":"2019","journal-title":"J. Inflammation"},{"key":"B36","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1533\/9780857093912.2.183","article-title":"Molecules involved in chemical defence of the chicken egg","author":"Rehault-Godbert","year":"2011","journal-title":"Improving the Safety and Quality of Eggs and Egg Products"},{"key":"B37","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1002\/jez.1400540207","article-title":"Effect of humidity on the growth, calcium metabolism, and mortality of the chick embryo.","volume":"54","author":"Romanoff","year":"1929","journal-title":"J. Exp. Zool."},{"key":"B38","author":"Romanoff","year":"1949","journal-title":"The Avian Egg."},{"key":"B39","doi-asserted-by":"publisher","first-page":"1828","DOI":"10.3382\/ps\/pev161","article-title":"Effect of Salmonella infection on cecal tonsil regulatory T cell properties in chickens.","volume":"94","author":"Shanmugasundaram","year":"2015","journal-title":"Poult. Sci."},{"key":"B40","doi-asserted-by":"publisher","first-page":"476","DOI":"10.1016\/j.vetimm.2011.08.016","article-title":"In vitro lipopolysaccharide treatment alters regulatory T cell properties in chickens.","volume":"144","author":"Shanmugasundaram","year":"2011","journal-title":"Vet. Immunol. Immunopathol."},{"key":"B41","doi-asserted-by":"publisher","first-page":"722","DOI":"10.3382\/ps\/pey460","article-title":"Effect of embryonic thermal manipulation on heat shock protein 70 (HSP70) expression and subsequent immune response to post-hatch lipopolysaccharide challenge in Pekin ducklings.","volume":"98","author":"Shanmugasundaram","year":"2019","journal-title":"Poult. Sci."},{"key":"B42","doi-asserted-by":"crossref","first-page":"354","DOI":"10.1006\/excr.1997.3476","article-title":"Hyperosmotic stress as a stimulant for proinflammatory cytokine production.","volume":"231","author":"Shapiro","year":"1997","journal-title":"Exp. Cell Res."},{"key":"B43","doi-asserted-by":"crossref","first-page":"326","DOI":"10.1016\/j.cell.2010.09.050","article-title":"SnapShot: network motifs.","volume":"143","author":"Shoval","year":"2010","journal-title":"Cell"},{"key":"B44","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1093\/icb\/20.2.385","article-title":"Water and ionic fluxes inside the egg.","volume":"20","author":"Simkiss","year":"1980","journal-title":"Am. Zool."},{"key":"B45","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1016\/0014-4827(69)90119-0","article-title":"Transport functions of the chick chorio-allantoic membrane: I. Normal histology and evidence for active electrolyte transport from the allantoic fluid, in vivo.","volume":"58","author":"Stewart","year":"1969","journal-title":"Exp. Cell Res."},{"key":"B46","first-page":"136","article-title":"The humidity factor in incubation","author":"Townsley","year":"1931","journal-title":"Proceedings of the 4th World\u2019s Poultry Congress at the Crystal Palace"},{"key":"B47","doi-asserted-by":"crossref","first-page":"529","DOI":"10.1016\/0300-9629(82)90118-9","article-title":"The relationship between eggshell porosity and oxygen consumption of the embryo in the domestic fowl.","volume":"72","author":"Tullett","year":"1982","journal-title":"Comp. Biochem. Physiol. A Physiol."},{"key":"B48","doi-asserted-by":"crossref","first-page":"1104","DOI":"10.1038\/ncb1071","article-title":"Rac\u2013MEKK3\u2013MKK3 scaffolding for p38 MAPK activation during hyperosmotic shock.","volume":"5","author":"Uhlik","year":"2003","journal-title":"Nat. Cell Biol."},{"key":"B49","doi-asserted-by":"crossref","first-page":"2145","DOI":"10.3382\/ps.2013-03006","article-title":"Effect of relative humidity during incubation at a set eggshell temperature and brooding temperature posthatch on embryonic mortality and chick quality.","volume":"92","author":"Van der Pol","year":"2013","journal-title":"Poult. Sci."},{"key":"B50","doi-asserted-by":"crossref","first-page":"397","DOI":"10.1186\/cc1546","article-title":"Hypertonic saline resuscitation in sepsis.","volume":"6","author":"Wade","year":"2002","journal-title":"Crit. Care"},{"key":"B51","doi-asserted-by":"crossref","first-page":"798","DOI":"10.3382\/ps.2008-00292","article-title":"Field study on broilers\u2019 first-week mortality.","volume":"88","author":"Yassin","year":"2009","journal-title":"Poult. Sci."},{"key":"B52","doi-asserted-by":"publisher","first-page":"1475","DOI":"10.1016\/S0002-9440(10)64423-9","article-title":"Lipopolysaccharide induces overexpression of MUC2 and MUC5AC in cultured biliary epithelial cells: possible key phenomenon of hepatolithiasis.","volume":"161","author":"Zen","year":"2002","journal-title":"Am. J. Pathol."}],"container-title":["Frontiers in Physiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fphys.2022.820349\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,3,7]],"date-time":"2022-03-07T08:49:20Z","timestamp":1646642960000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fphys.2022.820349\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,3,7]]},"references-count":52,"alternative-id":["10.3389\/fphys.2022.820349"],"URL":"https:\/\/doi.org\/10.3389\/fphys.2022.820349","relation":{},"ISSN":["1664-042X"],"issn-type":[{"value":"1664-042X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,3,7]]},"article-number":"820349"}}