Effect of dietary nonphytate phosphorus content on ileal lymphocyte subpopulations and cytokine expression in the cecal tonsils and spleen of laying hens that were or were not orally inoculated with Salmonella Typhimurium

Shi P. Bai Key Laboratory for Animal Disease-Resistance Nutrition of China's Ministry of Education, Institute of Animal Nutrition, Sichuan Agricultural University, Ya'an, Sichuan 625014, PR China.

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Yong Huang College of Veterinary Medicine, Sichuan Agricultural University, Ya'an Sichuan 625014, PR, China.

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Yu H. Luo Key Laboratory for Animal Disease-Resistance Nutrition of China's Ministry of Education, Institute of Animal Nutrition, Sichuan Agricultural University, Ya'an, Sichuan 625014, PR China.

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Lei L. Wang Key Laboratory for Animal Disease-Resistance Nutrition of China's Ministry of Education, Institute of Animal Nutrition, Sichuan Agricultural University, Ya'an, Sichuan 625014, PR China.

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Xue M. Ding Key Laboratory for Animal Disease-Resistance Nutrition of China's Ministry of Education, Institute of Animal Nutrition, Sichuan Agricultural University, Ya'an, Sichuan 625014, PR China.

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Jian P. Wang Key Laboratory for Animal Disease-Resistance Nutrition of China's Ministry of Education, Institute of Animal Nutrition, Sichuan Agricultural University, Ya'an, Sichuan 625014, PR China.

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Qiu F. Zeng Key Laboratory for Animal Disease-Resistance Nutrition of China's Ministry of Education, Institute of Animal Nutrition, Sichuan Agricultural University, Ya'an, Sichuan 625014, PR China.

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Ke Y. Zhang Key Laboratory for Animal Disease-Resistance Nutrition of China's Ministry of Education, Institute of Animal Nutrition, Sichuan Agricultural University, Ya'an, Sichuan 625014, PR China.

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Abstract

OBJECTIVE To evaluate the effects of dietary nonphytate phosphorus (NPP) content on ileal lymphocyte subpopulations and cytokine expression in the cecal tonsils and spleen of hens that were or were not inoculated with Salmonella Typhimurium.

ANIMALS 64 Salmonella-free hens.

PROCEDURES Hens were fed a diet with 0.22% (control; n = 32) or 0.42% (high-P; 32) NPP for 6 weeks and then orally inoculated with S Typhimurium (5 × 107 CFUs) or PBSS. Tissues were obtained from 8 S Typhimurium–inoculated and 8 PBSS-inoculated hens from each group at 2 and 7 days postinoculation (DPI). Percentages of ileal CD4+ and CD8+ lymphocytes were determined by flow cytometry. Cytokine mRNA expression was determined by quantitative real-time PCR assays.

RESULTS For S Typhimurium–inoculated hens, plasma parathyroid hormone concentration was significantly increased and 1,25-dihydroxyvitamin D3 concentration was decreased in hens fed the high-P diet, compared with values in hens fed the control diet. Salmonella Typhimurium inoculation caused an increase in the percentage of ileal CD8+ lymphocytes and the expression of interleukin (IL)-1β, IL-6, IL-8, interferon-γ, IL-12, and IL-18 in the cecal tonsils and spleen and a decrease in the expression of IL-4 and IL-10 in the cecal tonsils. Hens fed the high-P diet had significantly increased splenic expression of interferon-γ at 2 DPI and IL-1β, IL-6, IL-12, and IL-18 at 7 DPI, compared with hens fed the control diet.

CONCLUSIONS AND CLINICAL RELEVANCE Results suggested there was a T-helper 1 cytokine reaction in the cecal tonsils and spleen of S Typhimurium–inoculated hens, and dietary NPP content altered calcium regulation hormone concentrations and affected splenic cytokine expression.

Abstract

OBJECTIVE To evaluate the effects of dietary nonphytate phosphorus (NPP) content on ileal lymphocyte subpopulations and cytokine expression in the cecal tonsils and spleen of hens that were or were not inoculated with Salmonella Typhimurium.

ANIMALS 64 Salmonella-free hens.

PROCEDURES Hens were fed a diet with 0.22% (control; n = 32) or 0.42% (high-P; 32) NPP for 6 weeks and then orally inoculated with S Typhimurium (5 × 107 CFUs) or PBSS. Tissues were obtained from 8 S Typhimurium–inoculated and 8 PBSS-inoculated hens from each group at 2 and 7 days postinoculation (DPI). Percentages of ileal CD4+ and CD8+ lymphocytes were determined by flow cytometry. Cytokine mRNA expression was determined by quantitative real-time PCR assays.

RESULTS For S Typhimurium–inoculated hens, plasma parathyroid hormone concentration was significantly increased and 1,25-dihydroxyvitamin D3 concentration was decreased in hens fed the high-P diet, compared with values in hens fed the control diet. Salmonella Typhimurium inoculation caused an increase in the percentage of ileal CD8+ lymphocytes and the expression of interleukin (IL)-1β, IL-6, IL-8, interferon-γ, IL-12, and IL-18 in the cecal tonsils and spleen and a decrease in the expression of IL-4 and IL-10 in the cecal tonsils. Hens fed the high-P diet had significantly increased splenic expression of interferon-γ at 2 DPI and IL-1β, IL-6, IL-12, and IL-18 at 7 DPI, compared with hens fed the control diet.

CONCLUSIONS AND CLINICAL RELEVANCE Results suggested there was a T-helper 1 cytokine reaction in the cecal tonsils and spleen of S Typhimurium–inoculated hens, and dietary NPP content altered calcium regulation hormone concentrations and affected splenic cytokine expression.

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