Effects of deoxynivalenol contaminated corn distiller's dried grains with solubles on growth performance, body composition, immunological response, and gastrointestinal health in young pullets

Abstract

Mycotoxins, particularly deoxynivalenol (DON), are common contaminants in feed ingredients such as corn distiller's dried grains with solubles (DDGS) and pose significant risks to poultry health. This study investigated the effects of feeding naturally DON contaminated DDGS on growth performance, body composition, immunological response, and gastrointestinal health in young pullets. A total of 360, 4-week-old Hy-Line W36 pullets were randomly assigned to diets with increasing levels of naturally DON contaminated DDGS (0, 5, 10, 15, and 20%) over 28 days, resulting in dietary DON concentrations ranging from below the limit of quantification to 15.4 ppm. Diets with DON concentration exceeding 8.9 ppm, corresponding to 15% and 20% DDGS inclusion, resulted in significantly lower body weight gain (BWG) and feed intake (FI) from experimental day 14 to day 28 compared to DON concentration below 5.9 ppm (0, 5 and 10% DDGS groups; P = 0.024 and P = 0.007, respectively). Body composition analysis showed a higher tissue fat percentage in the 20% DDGS group (15.4 ppm DON) by day 28 compared to lower inclusion levels (P = 0.021). Immunologically, a significant increase in the CD4⁺:CD8⁺ ratio in spleen was observed in the 20% DDGS group compared to the 0% DDGS group (P = 0.013), whereas both 15 and 20% DDGS inclusion levels significantly increased the ratio in cecal tonsil (P < 0.001). Additionally, interleukin 1β (IL-1β) expression significantly increased in the cecal tonsil by day 28 with 15 and 20% DDGS inclusions (P = 0.002). Gut health was compromised as gut permeability increased linearly with increasing DDGS inclusion (linear, P = 0.043), aligning with significant alterations in the expression of the tight junction protein occludin (OCLN; P = 0.007). Antioxidant responses in the liver showed increased superoxide dismutase (SOD) activity in early exposure (day 13, P = 0.038), followed by decreased SOD activity (P = 0.001) and reduced glutathione (GSH) levels (P < 0.001) by day 28. In conclusion, feeding DON-contaminated DDGS at higher inclusion levels (15% and 20%) with final diet DON concentrations exceeding 8.9 ppm over 28 days adversely affects growth performance, immune function, and gut integrity in young pullets.

Keywords: Deoxynivalenol; Gastrointestinal health; Growth performance; Immune response; Pullets.

Fig. 1

T cell populations in spleen and cecal tonsils of pullets on day 13 fed diets with 0 to 20% DON-contaminated corn DDGS: A) CD4⁺ population percentage in the spleen; B) CD8⁺ population percentage in the spleen; C) CD4⁺:CD8⁺ ratio in the spleen; D) CD4⁺ population percentage in the cecal tonsil; E) CD8⁺ population percentage in the cecal tonsil; F) CD4⁺:CD8⁺ ratio in the cecal tonsil. Parameters are indicated by experimental day followed by bird age. Statistical analysis includes P-values from one-way ANOVA (P), linear regression (P_L), and quadratic regression (P_Q). Sample size: 6 tissues per treatment.

Fig. 2

T cell populations in spleen and cecal tonsils of pullets on day 28 fed diets with 0 to 20% DON-contaminated corn DDGS: A) CD4⁺ population percentage in the spleen; B) CD8⁺ population percentage in the spleen; C) CD4⁺:CD8⁺ ratio in the spleen; D) CD4⁺ population percentage in the cecal tonsil; E) CD8⁺ population percentage in the cecal tonsil; F) CD4⁺:CD8⁺ ratio in the cecal tonsil. Parameters are indicated by experimental day followed by bird age. Statistical analysis includes P-values from one-way ANOVA (P), linear regression (P_L), and quadratic regression (P_Q). Superscripts (a, b, c) denote statistically significant differences, with trendlines indicating linear and quadratic effects. Sample size: 6 tissues per treatment.

Fig. 3

Expression of inflammatory cytokines in cecal tonsils of pullets fed diets with 0 to 20% DON-contaminated corn DDGS over 28 days: A) Interleukin-1β mRNA expression on day 13; B) Interleukin-6 mRNA expression on day 13; C) Interleukin-10 mRNA expression on day 13; D) Interferon gamma mRNA expression on day 13; E) Interleukin-1β mRNA expression on day 28; F) Interleukin-6 mRNA expression on day 28; G) Interleukin-10 mRNA expression on day 28; H) Interferon gamma mRNA expression on day 28. Data are presented as fold changes using the 2^−ΔΔCt method. Parameters are indicated by experimental day followed by bird age. Statistical analysis includes P-values from one-way ANOVA (P), linear regression (P_L), and quadratic regression (P_Q). Superscripts (a, b, c) denote statistically significant differences, with trendlines indicating linear and quadratic effects. Sample size: 6 tissues per treatment.

Fig. 4

Antioxidant responses in pullets on day 13 fed diets with 0 to 20% DON-contaminated corn DDGS: A) Total Antioxidant Capacity (T-AOC) in blood serum; B) Superoxide Dismutase (SOD) activity in liver; C) Malondialdehyde (MDA) activity in liver; D) Reduced Glutathione (GSH) activity in liver; E) Oxidized Glutathione (GSSG) activity in liver; F) Total Glutathione (GSH+GSSG) activity in liver. Data are presented as activity levels. Parameters are indicated by experimental day followed by bird age. Statistical analysis includes P-values from one-way ANOVA (P), linear regression (P_L), and quadratic regression (P_Q). Superscripts (a, b) denote statistically significant differences, with trendlines indicating linear and quadratic effects. Sample size: 6 tissues per treatment.

Fig. 5

Antioxidant responses in pullets on day 28 fed diets with 0 to 20% DON-contaminated corn DDGS: A) Total Antioxidant Capacity (T-AOC) in blood serum; B) Superoxide Dismutase (SOD) activity in liver; C) Malondialdehyde (MDA) activity in liver; D) Reduced Glutathione (GSH) activity in liver; E) Oxidized Glutathione (GSSG) activity in liver; F) Total Glutathione (GSH+GSSG) activity in liver. Data are presented as activity levels. Parameters are indicated by experimental day followed by bird age. Statistical analysis includes P-values from one-way ANOVA (P), linear regression (P_L), and quadratic regression (P_Q). Superscripts (a, b) denote statistically significant differences, with trendlines indicating linear and quadratic effects. Sample size: 6 tissues per treatment.

Fig. 6

Gut permeability in pullets measured by FITC-d assay on days 13 and 28 of feeding diets with 0 to 20% DON-contaminated corn DDGS: A) Gut permeability on day 13; B) Gut permeability on day 28. FITC-d: fluorescein isothiocyanate-dextran. Statistical analysis includes P-values from one-way ANOVA (P), linear regression (P_L), and quadratic regression (P_Q). Trendline indicates linear effect. Sample size: 6 per treatment.

Fig. 7

Expression of tight junction proteins in the jejunum of pullets on days 13 and 28 of feeding diets with 0-20% DON-contaminated corn DDGS: A) Claudin-1 mRNA expression on day 13; B) Junction Adhesion Molecule-2 (JAM-2) mRNA expression on day 13; C) Occludin mRNA expression on day 13; D) Mucin-2 mRNA expression on day 13; E) Claudin-1 mRNA expression on day 28; F) Junction Adhesion Molecule-2 (JAM-2) mRNA expression on day 28; G) Occludin mRNA expression on day 28; H) Mucin-2 mRNA expression on day 28. Data are presented as fold changes using the 2^−ΔΔCt method. Parameters are indicated by experimental day followed by bird age. Statistical analysis includes P-values from one-way ANOVA (P), linear regression (P_L), and quadratic regression (P_Q). Superscripts (a, b) denote statistically significant differences, with trendlines indicating linear and quadratic effects. Sample size: 6 tissues per treatment.