Amelioration of Ochratoxin A-induced immunotoxic effects by silymarin and vitamin E in White Leghorn cockerels

Figures & data

Table 1.  Layout of the experiment.

Groups	No. of birds	Treatment in feed
Control	20	Nil
OTA1	20	OTA (1 mg/kg)
OTA2	20	OTA (2 mg/kg)
Silymarin (SL)	20	Silymarin (10 g/kg)
Vitamin E (VE)	20	Vitamin E (200 mg/kg)
OTA1+SL	20	OTA (1 mg/kg) + Silymarin (10 g/kg)
OTA2+SL	20	OTA (2 mg/kg) + Silymarin (10 g/kg)
OTA1+VE	20	OTA (1 mg/kg) + Vitamin E (200 mg/kg)
OTA2+VE	20	OTA (2 mg/kg) + Vitamin E (200 mg/kg)
VE+SL	20	Vitamin E (200 mg/kg) + Silymarin (10 g/kg)
OTA1+SL+VE	20	OTA (1 mg/kg) + Silymarin (10 g/kg) + Vitamin E (200 mg/kg)
OTA2+SL+VE	20	OTA (2 mg/Kg) + Silymarin (10 g/kg) + Vitamin E (200 mg/kg)

OTA1, group receiving 1 mg/kg OTA in feed; OTA2, group receiving 2 mg/kg OTA in feed; SL, Silymarin; VE, Vitamin E.

Table 2.  Antibody titers (log2) against SRBC at different days.

Groups	Time after primary SRBC injection				Time after booster SRBC injection
	7 days		14 days		7 days		14 days
	Total Ig	Ig G	Total Ig	Ig G	Total Ig	Ig G	Total Ig	Ig G
Control	2.99 ± 0.21^a	2.51 ± 0.23^a	2.70 ± 0.13^a	2.24 ± 0.16^a	2.86 ± 0.10^a	2.45 ± 0.15^a	2.90 ± 0.11^a	2.59 ± 0.00^a
OTA1	2.31 ± 0.24^cd	1.98 ± 0.30^ab	1.59 ± 0.00^c	1.43 ± 0.29^bc	2.31 ± 0.24^b	2.08 ± 0.16^bc	2.45 ± 0.15^bc	2.00 ± 0.01^bcd
OTA2	2.31 ± 0.24^cd	1.95 ± 0.43^ab	1.00 ± 0.00^d	0.75 ± 0.50^d	1.90 ± 0.21^e	1.69 ± 0.21^d	1.98 ± 0.30^de	1.44 ± 0.29^f
Silymarin (SL)	2.80 ± 0.17^ab	2.37 ± 0.28^a	2.52 ± 0.13^a	1.90 ± 0.21^abc	2.85 ± 0.20^a	2.45 ± 0.15^a	2.85 ± 0.20^a	2.31 ± 0.24^ab
Vitamin E (VE)	2.99 ± 0.15^a	2.58 ± 0.20^a	2.51 ± 0.23^a	2.06 ± 0.35^ab	2.85 ± 0.20^a	2.45 ± 0.15^a	2.70 ± 0.13^ab	2.23 ± 0.28^abc
OTA1+SL	2.80 ± 0.17^ab	2.39 ± 0.13^a	2.68 ± 0.29^a	2.23 ± 0.28^a	2.85 ± 0.20^a	2.37 ± 0.28^ab	2.80 ± 0.17^a	2.37 ± 0.28^ab
OTA2+SL	1.65 ± 0.47^e	1.04 ± 0.75^c	1.79 ± 0.24^bc	1.69 ± 0.21^bc	2.31 ± 0.24^b	2.08 ± 0.16^bc	2.24 ± 0.28^cd	1.87 ± 0.36^cde
OTA1+VE	2.52 ± 0.13^bc	2.08 ± 0.16^a	2.59 ± 0.01^a	2.16 ± 0.19^a	2.85 ± 0.20^a	2.31 ± 0.24^abc	2.74 ± 0.20^ab	2.39 ± 0.13^ab
OTA2+VE	1.98 ± 0.30^de	1.44 ± 0.29^bc	1.69 ± 0.21^bc	1.40 ± 0.49^c	2.31 ± 0.24^b	1.98 ± 0.30^cd	2.08 ± 0.16^de	1.79 ± 0.24^def
VE+SL	2.67 ± 0.13^abc	2.23 ± 0.28^a	2.58 ± 0.20^a	2.12 ± 0.43^a	2.80 ± 0.17^a	2.31 ± 0.24^abc	2.64 ± 0.11^ab	2.08 ± 0.16^bcd
OTA1+SL+VE	2.80 ± 0.17^ab	2.39 ± 0.13^a	2.75 ± 0.11^a	2.23 ± 0.28^a	2.86 ± 0.10^a	2.51 ± 0.23^a	2.80 ± 0.17^a	2.37 ± 0.28^ab
OTA2+SL+VE	1.65 ± 0.47^e	1.04 ± 0.75^e	1.90 ± 0.21^b	1.44 ± 0.29^bc	2.24 ± 0.16^b	1.98 ± 0.30^cd	1.90 ± 0.21^e	1.54 ± 0.41^ef

Values shown are mean (± SD) from n = 5 birds/group at each timepoint.

Values in each column followed by different letters are significantly different at p ≤ 0.05.

OTA, Ochratoxin A; SL, Silymarin; VE, Vitamin E. All other designations are as outlined in footer of Table 1.

Table 3.  Lymphoproliferative response (mm) to subcutaneous injection of avian tuberculin.

Groups	24 h post-injection	48 h post-injection	72 h post-injection
Control	0.14 ± 0.03^a	0.10 ± 0.07	0.03 ± 0.03
OTA1	0.06 ± 0.04^bc	0.09 ± 0.07	0.08 ± 0.07
OTA2	0.05 ± 0.02^bc	0.06 ± 0.05	0.09 ± 0.08
Silymarin (SL)	0.11 ± 0.04^ab	0.03 ± 0.03	0.08 ± 0.05
Vitamin E (VE)	0.12 ± 0.03^a	0.09 ± 0.07	0.07 ± 0.04
OTA1+SL	0.14 ± 0.05^a	0.057 ± 0.02	0.06 ± 0.01
OTA2+SL	0.07 ± 0.03^bc	0.03 ± 0.04	0.07 ± 0.06
OTA1+VE	0.02 ± 0.01^c	0.05 ± 0.04	0.08 ± 0.03
OTA2+VE	0.03 ± 0.01^c	0.04 ± 0.03	0.05 ± 0.04
VE+SL	0.14 ± 0.03^a	0.04 ± 0.05	0.07 ± 0.02
OTA1+SL+VE	0.09 ± 0.04^ab	0.03 ± 0.03	0.02 ± 0.02
OTA2+SL+VE	0.05 ± 0.05^bc	0.09 ± 0.07	0.07 ± 0.07

Values shown are mean (± SD) from n = 5 birds/group at each timepoint.

Values in each column followed by different letters are significantly different at p ≤ 0.05.

OTA, Ochratoxin A; SL, Silymarin; VE, Vitamin E. All other designations are as outlined in footer of Table 1.

Table 4.  Phagocytic index as determined by carbon clearance assay.

Groups	At 3 min	At 15 min
Control	0.45 ± 0.08^ab	0.06 ± 0.03^a
OTA1	0.30 ± 0.06^c	0.04 ± 0.01^c
OTA2	0.30 ± 0.06^c	0.03 ± 0.02^cd
Silymarin (SL)	0.44 ± 0.02^ab	0.05 ± 0.01^ab
Vitamin E (VE)	0.41 ± 0.06^ab	0.07 ± 0.02^a
OTA1+SL	0.24 ± 0.06^bc	0.05 ± 0.02^ab
OTA2+SL	0.13 ± 0.08^c	0.02 ± 0.02^de
OTA1+VE	0.61 ± 0.26^a	0.06 ± 0.01^a
OTA2+VE	0.26 ± 0.15^bc	0.02 ± 0.03^de
VE+SL	0.38 ± 0.05^b	0.06 ± 0.01^ab
OTA1+SL+VE	0.34 ± 0.01^bc	0.05 ± 0.01^ab
OTA2+SL+VE	0.11 ± 0.03^c	0.01 ± 0.01^ef

Values shown are mean (± SD) from n = 5 birds/group at each timepoint.

Values were determined as K = (log_n OD₁ − log_n OD₂)/(T₂ − T₁), where OD₁ and OD₂ are optical densities (640 nm) at times T₁ and T₂, respectively. Absorbance from a ‘0 min’ sample was considered as the ‘zero’ value for each respective sample.

Values in each column followed by different letters are significantly different at p ≤0.05.

OTA, Ochratoxin A; SL, Silymarin; VE, Vitamin E. All other designations are as outlined in footer of Table 1.