Use of encapsulated Bacillus subtilis and essential oils to improve antioxidant and immune status of blood and production and hatching performance of laying hens

Figures & data

Table 1. Ingredients and chemical composition of the experimental basal diets (dry matter basis).

Ingredients^a (%, unless otherwise indicated)	Content
Corn	62.00
Soybean meal	26.00
Limestone	5.50
CaHPO4	1.00
Premix^b	5.00
Total	100.00
Calculated composition
ME, MJ/kg	11.18
CP, %	16.14
Ca, %	2.99
AP, %	0.38
Salt, %	0.37
Lys, %	0.95
Met, %	0.51
Met + Cys, %	0.82

^aCaHPO₄, Calcium hydrogen phosphate; ME, metabolisable energy; CP, crude protein; Ca, Calcium; AP, available phosphorus; Lys, lysine; Met, methionine; Met + Cys, methionine + cysteine; BSEO, mixture of Bacillus subtilis and essential oils.

^bPremix provided per kilogram of diet: vitamin A, 170 000 IU; vitamin D3, 64 000 IU; vitamin E, 880 IU; vitamin B1, 48 mg; vitamin B2, 105 mg; vitamin B6, 48 mg; vitamin B12, 0.20 mg; vitamin K3, 48 mg; nicotinic acid, 380 mg; pantothenic acid, 270 mg; folic acidm, 24 mg; Zn (from zinc sulfate), 1560 mg; Fe (from ferrous sulfate), 1100 mg; Mn (from manganese sulfate), 1800 mg; Cu (from copper sulfate), 240 mg; I (from potassium iodide) 23 mg; Se (from sodium selenite), 4.80 mg; Ca, 10.00 g; P, 2.60 g; NaCl, 3.70 g.

Table 2. Effects of increasing inclusion of dietary BSEO on production performance of laying hens*.

Item	Dietary treatment**				Pooled SEM	p-value
	CON	BSEO-1	BSEO-2	BSEO-3		ANOVA	Linear	Quadratic
Egg weight (g)	44.5	44.4	44.4	44.8	0.3	.966	.714	.728
Egg production (%)	65.3	65.6	66.0	68.0	0.4	.076	.020	.278
Feed intake (g/d per hen)	88.8	87.9	88.1	88.0	1.0	.989	.818	.832
Feed conversion ratio (g of feed/g of e.g. g)	3.48	3.21	3.18	3.04	0.06	.079	.014	.607

*Data are means of 6 replicates per treatment with 32 hens per replicate. **CON, BSEO-1, BSEO-2, BSEO-3 diets contained 0, 300, 600, 900 mg/kg of BSEO, respectively.

^a–bMeans within a row with different superscripts differ significantly (p < .05).

Table 3. Effects of increasing inclusion of dietary BSEO on hatching performance of laying hens*.

Item	Dietary treatment**				Pooled SEM	p-value
	CON	BSEO-1	BSEO-2	BSEO-3		ANOVA	Linear	Quadratic
Fertility (%)	95.4	96.0	96.2	95.9	0.2	.417	.234	.235
Hatchability (%)	84.5^b	87.3^a	86.7^a	87.0^a	0.3	.001	.004	.017
Hatchability of fertile e.g. gs (%)	88.7^b	91.0^a	90.3^ab	90.7^a	0.2	.002	.010	.038

*Data are means of 6 replicates per treatment with 32 hens per replicate.

**CON, BSEO-1, BSEO-2, BSEO-3 diets contained 0, 300, 600, 900 mg/kg of BSEO, respectively.

Table 4. Effect of increasing inclusion of dietary BSEO on egg quality of laying hens*.

Item	Dietary treatment**				Pooled SEM	p-value
	CON	BSEO-1	BSEO-2	BSEO-3		ANOVA	Linear	Quadratic
Eggshell-breaking strength (kgf)	4.06	4.05	3.82	3.90	0.07	.558	.265	.721
Eggshell thickness (mm)	0.333	0.334	0.329	0.319	0.002	.096	.032	.203
Yolk index	0.377^b	0.386^ab	0.392^ab	0.397^a	0.002	.007	.001	.476
Yolk colour	5.55	5.30	5.29	5.75	0.16	.576	.623	.200
Haugh unit	60.5	60.9	61.0	61.5	0.7	.971	.636	.988

*Data are means of 6 replicates per treatment with 32 hens per replicate.

**CON, BSEO-1, BSEO-2, BSEO-3 diets contained 0, 300, 600, 900 mg/kg of BSEO, respectively.

^a–bMeans within a row with different superscripts differ significantly (p < .05).

Table 5. Effect of increasing inclusion of dietary BSEO on serum antioxidant capacity of laying hens*.

Item**	Dietary treatment***				Pooled SEM	p-value
	CON	BSEO-1	BSEO-2	BSEO-3		ANOVA	Linear	Quadratic
GSH-Px (U/mL)	2271^b	2580^ab	2756^a	2795^a	58	.002	<.001	.168
T-SOD (U/mL)	228	235	238	234	1	.220	.167	.115
T-AOC (U/mL)	5.17^c	6.26^bc	7.67^ab	7.76^a	0.26	<.001	<.001	.184
MDA (nmol/mL)	6.29	5.61	5.50	6.05	0.20	.451	.639	.126

*Data are means of 2 hens of 6 replicates per treatment.

**GSH-Px, glutathione peroxidase; T-SOD, total superoxide dismutase; T-AOC, total antioxidant capacity; MDA, malondialdehyde.

***CON, BSEO-1, BSEO-2, BSEO-3 diets contained 0, 300, 600, 900 mg/kg of BSEO, respectively.

^a–cMeans within a row with different superscripts differ significantly (p < .05).

Table 6. Effects of increasing inclusion of dietary BSEO on serum immune indexes of laying hens*.

Item**	Dietary treatment***				Pooled SEM	p-value
	CON	BSEO-1	BSEO-2	BSEO-3		ANOVA	Linear	Quadratic
IgA (g/L)	2.24	2.20	2.17	2.21	0.02	.332	.318	.144
IgG (g/L)	4.21	4.11	4.04	4.11	0.03	.157	.133	.093
IgM (g/L)	1.64	1.60	1.58	1.61	0.01	.237	.284	.084
NDV-Ab	1.14	1.21	1.28	1.10	0.05	.680	.896	.240
AIV-Ab	0.960^b	1.24^a	1.16^a	1.22^a	0.03	.003	.004	.042

*Data are means of 2 hens of 6 replicates per treatment. **IgA, immunoglobulin A; IgG, immunoglobulin G; IgM, immunoglobulin M; NDV-Ab, Newcastle disease virus antibody; AIV-Ab, avian influenza virus antibody. ***CON, BSEO-1, BSEO-2, BSEO-3 diets contained 0, 300, 600, 900 mg/kg of BSEO, respectively.

^a–bMeans within a row with different superscripts differ significantly (p < .05).

Table 7. Effect of increasing inclusion of dietary BSEO on serum hormone index of laying hens*.

Item**	Dietary treatment***				Pooled SEM	p-value
	CON	BSEO-1	BSEO-2	BSEO-3		ANOVA	Linear	Quadratic
E2 (pg/mL)	16.0^c	17.9^b	16.0^c	19.5^a	0.3	<.001	<.001	.059
PTH (pg/mL)	144^c	158^ab	163^a	152^b	2	<.001	.003	<.001
P (ng/mL)	1.80	1.60	1.64	1.77	0.07	.711	.970	.261
LH/FSH	1.13^b	1.22^ab	1.18^ab	1.37^a	0.03	.047	.016	.393

*Data are means of 2 hens of 6 replicates per treatment. **E₂, oestradiol; PTH, parathyroid hormone; P, progesterone; LH, luteotropic hormone; FSH, follicle-stimulating hormone. ***CON, BSEO-1, BSEO-2, BSEO-3 diets contained 0, 300, 600, 900 mg/kg of BSEO, respectively.

^a–cMeans within a row with different superscripts differ significantly (p < .05).