Protective role of bentonite against aflatoxin B₁- and ochratoxin A-induced immunotoxicity in broilers

Figures & data

Table 1. Chemical composition of bentonite clay (BN).

Chemical composition (w/w %)
Loss on ignition (L/I)	14.00
Silica (SiO2)	46.28
Alumina (AL2O3)	14.70
Iron Oxide (Fe2O3)	7.50
Calcium Oxide (CaO)	11.22
Magnesium Oxide (MgO)	5.00
Sodium Oxide (Na2O)	0.91
Potassium Oxide (K2O)	0.38
Specific Gravity (unitless)	2.24
Liquid Limit (%)	40.00
Plastic Limit (%)	24.00
Plasticity Index (unitless)	16.00

Table 2. Treatment regimens.

Groups	No of birds(in each group)	Treatment
Experiment 1; Dietary AFB1 (mg/g feed) and/or BN (g/kg feed)
Control	30	AFB1 0, BN 0
A1, A2, A3	30	AFB1: 0.1, 0.2, 0.6
B1, B2	30	BN: 3.7, 7.5
A1B1, A2B1, A3B1	30	AFB1: 0.1, 0.2, 0.6 BN: 3.7 with each AFB1 level
A1B2, A2B2, A3B2	30	AFB1: 0.1, 0.2, 0.6 BN: 7.5 with each AFB1 level
Experiment 2. Dietary OTA (mg/g feed) and/or BN (g/kg feed)
Control	30	OTA 0, BN 0
O1, O2, O3	30	OTA: 0.15, 0.3, 1.0
B1, B2, B3	30	BN: 3.7, 7.5, 15
O1B1, O2B1, O3B1	30	OTA: 0.15, 0.3, 1.0 BN 3.7 with each OTA level
O1B2, O2B2, O3B2	30	OTA: 0.15, 0.3, 1.0 BN: 7.5 with each OTA level
O1B3, O2B3, O3B3	30	OTA: 0.15, 0.3, 1.0 BN: 15.0 with each OTA level

Figure 1. (A–D) Antibodies titers against SRBC in broiler chicks given different dietary levels of AFB₁ and bentonite clay. Values shown are mean [± SD] titers (log₃ of reciprocal of highest dilution giving visible agglutination, n = 6/group) Value significantly different from control at *p < 0.05, or **p < 0.01. Abbreviations: A1 = 0.1 mg AFB₁/kg feed, A2 = 0.2 mg AFB₁/kg feed A3 = 0.6 mg AFB₁/kg feed, B1 = 3.7 g bentonite clay/kg feed, B2 = 7.5 g bentonite clay/kg feed. Bars (left to right, each set): control, A1, A2, A3, B1, B2, A1B1, A2B1, A3B1, A1B2, A2B2, A3B2.

Figure 2. (A, B). In-vivo phagocytic potential, of broiler chicks fed different dietary levels of AFB₁ and/or bentonite clay. Bars (left to right, each set): control, A1, A2, A3, B1, B2, A1B1, A2B1, A3B1, A1B2, A2B2, A3B2. Values shown are means ± SD (n = 6 chicks/group). Value significantly different from control at *p < 0.05, **p < 0.01, or ***p < 0.001. Abbreviations are as reported in Figure 1.

Figure 3. (A–C) Lymphoproliferative response to PHA-P of chicks fed different dietary levels of AFB₁ and/or bentonite clay. Bars (left to right, each set): control, A1, A2, A3, B1, B2, A1B1, A2B1, A3B1, A1B2, A2B2, A3B2. Values shown are means ± SD (n = 6 chicks/group). Value significantly different from control at *p < 0.05 or **p < 0.01. Abbreviations are as reported in Figure 1.

Figure 4. Follicular diameter of bursa of Fabricius of chicks fed different dietary levels of AFB₁ and/or bentonite clay. Values shown are means ± SD (n = 6 chicks/group). Value significantly different from control at *p < 0.05 or **p < 0.01. Abbreviations are as reported in Figure 1.

Figure 5. Intermolecular connective tissue of bursa of Fabricius of chicks fed different dietary levels of AFB₁ and/or bentonite clay. Values shown are means ± SD (n = 6 chicks/group). Value significantly different from control at *p < 0.05 or **p < 0.01. Abbreviations are as reported in Figure 1.

Figure 6. (A–C) Antibody titers against SRBC in broiler chicks given different dietary levels of OTA and bentonite clay. Values shown are mean [± SD titers] (log₃ of reciprocal of highest dilution giving visible agglutination, n = 6/group) Value significantly different from control at *p < 0.05, or **p < 0.01. Abbreviations: O1 = 0.15 mg OTA/kg feed, O2 = 0.3 mg OTA/kg feed, O3 = 1.0 mg OTA/kg feed, B1 = 3.7 g bentonite clay/kg, B2 = 7.5 g bentonite clay/kg feed, B3 = 15 g bentonite clay/kg feed. Bars (left to right, each set): control, O1, O2, O3, B1, B2, B3, O1B1, O2B1, O3B1, O1B2, O2B2, O3B2, O1B3, O2B3, O3B3.

Figure 7. (A, B) In vivo phagocytic potential of chicks fed different dietary levels of OTA and/or bentonite clay. Bars (left to right, each set): control, O1, O2, O3, B1, B2, B3, O1B1, O2B1, O3B1, O1B2, O2B2, O3B2, O1B3, O2B3, O3B3. Abbreviations are as reported in Figures 6. Values shown are means ± SD (n = 6 chicks/group). Value significantly different from control at *p < 0.05, **p < 0.01, or ***p < 0.001.

Figure 8. (A–C) Lymphoproliferative response to PHA-P of chicks fed different dietary levels of OTA and/or bentonite clay. Bars (left to right, each set): control, O1, O2, O3, B1, B2, B3, O1B1, O2B1, O3B1, O1B2, O2B2, O3B2, O1B3, O2B3, O3B3. Abbreviations are as reported in Figures 6. Values shown are means ± SD (n = 6 chicks/group). Value significantly different from control at *p < 0.05.

Figure 9. Follicular diameter of bursa of Fabricius of broiler chicks fed different dietary levels of OTA and/or bentonite clay. Values shown are means ± SD (n = 6 chicks/group). Value signifi-cantly different from control at *p < 0.05 or **p < 0.01. Abbreviations are as reported in Figures 6.

Figure 10. Inter-follicular connective tissue (μm) of bursa of Fabricius of chicks fed different dietary levels of OTA and/or bentonite clay. Values shown are means ± SD (n = 6 chicks/group). Value significantly different from control at *p < 0.05 or **p < 0.01. Abbreviations are as reported in Figures 6.