Effects of Dietary Zinc on Performance, Zinc Transporters Expression, and Immune Response of Aged Laying Hens

  • Qiqi Han
  • Yuming Guo
  • Bingkun Zhang
  • Wei NieEmail author


This study was to investigate the effects of dietary zinc (Zn) supplementation on performance, zinc transporter gene expression, and immune function in aged laying hens. In experiment 1, twenty 31-week-old hens (young) and twenty 60-week-old hens (old) with the same genetic background were fed with the same diet for 4 weeks. In experiment 2, a basal diet supplemented with zinc sulfate (ZnS) and zinc glycine chelate (ZnG) at 30, 60, 90, and 120 mg Zn/kg to constitute nine experimental diets. Eight hundred and ten 60-week-old layers were distributed in a completely randomized experimental design with 9 treatments, 6 replicates of 15 birds each, and birds were fed for 10 weeks. In experiment 1, results showed that zinc and metallothionein (MT) concentration in the shell gland of old hens was significantly lower than young layers (P < 0.05). Zinc transporters ZnT1, 4, 5, 6, and 7 messenger RNA (mRNA) abundance in old layers were significantly lower versus the young (P < 0.05). In experiment 2, results indicated that dietary zinc supplementation did not significantly affect the laying rate, average feed intake, egg weight, feed conversion efficiency, broken egg rate, or mortality (P > 0.05). Supplemental ZnG significantly improved eggshell breaking strength than ZnS, with a higher alkaline phosphatase (ALP) activity and more abundant ZnT4 expression in shell gland versus ZnS (P < 0.05). ZnG supplementation at 90 mg Zn/kg affected the duodenal mucus by significantly increasing ZnT1, 6, 7, ZIP13, and MT-4 mRNA level (P < 0.05). Zinc level significantly increased bovine serum albumin (BSA) antibody concentration on 14 day after BSA injection (P < 0.05). Supplementation of ZnG improved eggshell quality of aged layers by upgrading zinc transporter expression in the shell gland and intestine also enhanced humoral immunity.


Zinc Laying hen Shell gland Transporter Immune 



alkaline phosphatase


bovine serum albumin




enzyme-linked immunosorbent assay


endoplasmic reticulum


graphite furnace atomic absorption spectrometry


messenger RNA




metal-response element-binding transcription factor-1


quantitative real-time polymerase chain reaction


solute-linked carrier


trans-Golgi network


Zrt- and Irt-like proteins




zinc glycine chelate


zinc sulfate


Zinc transporter



The authors would like to thank the staff of the Department of Animal Science and Technology of the China Agricultural University for their valuable assistance in sample collecting. The authors are also grateful to Dr. Kenny Ray Hazen for his linguistic help.

Authors’ Contributions

Qiqi Han performed the animal experiment, established sample and statistical analysis and wrote the manuscript; Yuming Guo designed the research, interpreted the data; Bingkun Zhang assisted with the design of the research and data analysis; Wei Nie adjusted the manuscript writing and experiment designing; all authors read and approved the final manuscript.


The State Key Development Program (2016YFD0501202) supported this study.

Compliance with Ethical Standards

Ethics Approval and Consent to Participate

All procedures of animal care and use for this experiment were approved by China Agricultural University Institutional Animal Care and Use Committee (No. CAU20160910-2).

Consent for Publication

Not applicable.

Competing Interests

The authors declare that they have no competing interests.


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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.The State Key Laboratory of Animal Nutrition, College of Animal Science and TechnologyChina Agricultural UniversityBeijingPeople’s Republic of China

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