Effects of dietary Bacillus coagulans on the productive performance, egg quality, serum parameters, and intestinal morphology of laying hens during the late laying period

Figures & data

Table 1. Composition and nutrient levels of the experimental basal diet for laying hens.

Ingredients (% on dry matter basis)	Basal diet
Corn	63
Soybean meal	26
Limestone	8
DL-Methionine (98%)	0.1
CaHPO4	0.9
Premix^a	2
Total	100
Calculated nutritional level (%)
Metabolisable energy (MJ/kg)	2.65
Analysed nutritional level (%)
Crude protein	16.59
Calcium	3.23
Phosphorus	0.54
Methionine	0.36
Lysine	0.85
Threonine	0.63
Tryptophan	0.19
Arginine	1.11
Valine	0.76

^aPremix provided the following per kilogram of diet: vitamin A, 4000 IU; vitamin D3, 1200 IU; vitamin E, 6 mg; vitamin B1, 1.4 mg; vitamin B2, 3 mg; vitamin B6, 1.0 mg; vitamin B12, 0.01 mg; pantothenic acid, 7.5 mg; choline chloride, 500 mg; biotin, 0.15 mg; Ca,7500 mg; P, 3000 mg; Mn, 72 mg; Zn, 56 mg; Fe, 60 mg; Cu, 25 mg; I, 0.50 mg; Se, 0.10 mg; phytase, 2000 FTU (phytase unit).

Table 2. Effects of B. coagulans on productive performance of laying hens (n = 32).

Item	BC supplementation (g/t)					SEM	p-Value	PL	PQ
	0	50	100	150	200
Egg mass, g/hen/d	51.73^b	51.87^b	55.29^a	52.18^ab	55.01^ab	0.12	0.002	0.012	0.039
Laying rate, %	80.21^b	80.10^b	83.71^a	81.19^b	83.16^a	0.07	<0.001	<0.001	<0.001
Egg weight, g	64.76^b	65.09^b	66.03^a	65.15^b	65.03^b	0.18	<0.001	0.135	<0.001
Feed intake, g/hen/d	105.04	104.29	105.37	104.57	106.10	0.80	0.185	0.184	0.187
FCR, kg feed/kg egg	2.06^a	2.05^a	1.94^b	2.04^a	2.07^a	0.03	<0.001	0.796	<0.001
Breaking rate, %	4.73	4.36	3.81	4.14	3.96	0.31	0.050	0.018	0.020

^a,bMeans sharing different letters in the same row are significantly different (p < 0.05).

BC: Bacillus coagulans; FCR: feed conversion ratio; SEM: standard error of the means; PL: p-Value of linear analysis; PQ: p-Value of quadratic analysis.

Table 3. Effects of B. coagulans on egg quality of laying hens (n = 30).

Item	BC supplementation (g/t)					SEM	p-Value	PL	PQ
	0	50	100	150	200
Albumen height, mm	6.25	6.28	6.13	6.23	6.10	0.26	0.941	0.529	0.817
Haugh unit	76.72	76.01	75.48	75.08	74.81	2.03	0.889	0.292	0.565
Egg yolk colour, points	6.00	6.00	6.00	6.00	6.20	0.15	0.603	0.241	0.308
Eggshell strength, kgf/m^2	3.71	3.74	4.11	3.79	3.91	0.25	0.509	0.418	0.530
Eggshell thickness, mm	0.45	0.46	0.45	0.45	0.44	0.01	0.789	0.342	0.548

BC: Bacillus coagulans; SEM: standard error of the means; PL: p-Value of linear analysis; PQ: p-Value of quadratic analysis. Serum biochemistry.

Table 4. Effects of B. coagulans on serum biochemistry of laying hens (n = 12).

Item	BC supplementation (g/t)					SEM	p-Value	PL	PQ
	0	50	100	150	200
ALT, U/L	19.65	14.91	16.50	17.21	16.14	0.90	0.608	0.439	0.556
AST, U/L	23.84	17.98	22.62	16.91	15.56	1.60	0.425	0.123	0.312
TP, g/L	44.06^b	57.06^ab	59.54^a	46.46^ab	48.18^ab	5.21	0.020	0.941	0.050
ALB, g/L	26.27	18.79	20.13	22.17	22.12	3.00	0.138	0.471	0.115
ALP, U/L	26.61^ab	26.38^ab	15.29^b	18.54^ab	33.59^a	5.82	0.031	0.822	0.027
TC, mmol/L	0.53	0.47	0.46	0.43	0.59	0.09	0.483	0.844	0.242
TG, mmol/L	23.56	17.24	15.50	18.29	22.06	4.48	0.354	0.906	0.112
Ca, mmol/L	1.67	1.52	1.29	1.22	1.65	0.19	0.061	0.501	0.028
P, mmol/L	0.72	0.54	0.83	0.70	0.78	0.15	0.262	0.295	0.453
UA, μmol/L	1807.83	1423.06	1785.06	1869.88	2043.37	434.28	0.630	0.314	0.411

^a,bMeans sharing different letters in the same row are significantly different (p < 0.05).

ALT: glutamic-pyruvic transaminase; AST: glutamic-oxalacetic transaminase; TP: total protein; ALB: albumin; ALP: alkaline phosphatase; TC: total cholesterol; TG: triglyceride; Ca: calcium; P: phosphorus; UA: Uric acid; BC: Bacillus coagulans; SEM: standard error of the means; PL: p-Value of linear analysis; PQ: p-Value of quadratic analysis.

Table 5. Effects of B. coagulans on serum hormone and antioxidant enzyme activities of laying hens (n = 12).

Item	BC supplementation (g/t)					SEM	p-Value	PL	PQ
	0	50	100	150	200
LH, ng/L	26.66	25.95	22.18	27.10	31.05	4.37	0.285	0.321	0.131
FSH, U/L	6.17	6.18	5.25	5.93	5.66	0.76	0.557	0.375	0.560
E2, pmol/L	40.28	38.72	34.67	41.63	45.21	0.63	0.235	0.281	0.109
T-AOC, U/mL	1.55	1.58	1.56	1.52	1.62	0.12	0.936	0.818	0.902
SOD, U/mL	20.97	21.63	22.38	22.63	21.02	1.15	0.488	0.676	0.232
CAT, U/mL	10.52	12.64	6.63	8.04	9.85	3.89	0.211	0.298	0.412
GSH-Px, U/mL	335.89^b	397.50^a	407.68^a	385.71^ab	364.29^ab	20.91	0.012	0.402	0.003
MDA, nmol/mL	52.03	33.22	27.21	49.04	50.58	12.61	0.398	0.607	0.256

^a,bMeans sharing different letters in the same row are significantly different (p < 0.05).

LH: luteinising hormone; FSH: follicle-stimulating hormone; E₂: oestradiol; T-AOC: total antioxidant capacity; SOD: superoxide dismutase; CAT: catalase; GSH-Px: glutathione peroxide; MDA: malondialdehyde; BC: Bacillus coagulans; SEM: standard error of the means; PL: p-Value of linear analysis; PQ: p-Value of quadratic analysis. Hepatic antioxidant parameters.

Table 6. Effects of B. coagulans on hepatic antioxidant enzyme activities of laying hens (n = 12).

Item	BC supplementation (g/t)					SEM	p-Value	PL	PQ
	0	50	100	150	200
T-AOC, U/mg prot	0.85	0.83	0.84	0.83	0.83	0.01	0.119	0.088	0.086
CAT, U/ mg prot	13.79^ab	16.68^ab	12.63^b	11.97^b	18.99^a	3.09	0.007	0.289	0.129
GSH-Px, U/mg prot	685.55	593.92	687.08	566.23	644.04	58.89	0.588	0.599	0.813
MDA, nmol/mg prot	6.16	4.90	5.40	4.54	4.65	1.69	0.824	0.311	0.567

^a,bMeans sharing different letters in the same row are significantly different (p < 0.05).

T-AOC: total antioxidant capacity; CAT: catalase; GSH-Px: glutathione peroxide MDA: malondialdehyde; BC: Bacillus coagulans; SEM: standard error of the means; PL: p-Value of linear analysis; PQ: p-Value of quadratic analysis.

Table 7. Effects of B. coagulans on immune response in laying hens (n = 12).

Item	BC supplementation (g/t)					SEM	p-Value	PL	PQ
	0	50	100	150	200
IgA, µg/mL	3.32^b	3.62^ab	3.22^b	3.75^ab	4.70^a	0.33	0.027	0.011	0.009
IgG, µg/mL	23.02	20.38	13.78	12.10	20.25	2.42	0.055	0.266	0.030
IgM, µg/mL	3.51	3.27	2.37	2.55	2.33	0.37	0.103	0.012	0.033

^a,bMeans sharing different letters in the same row are significantly different (p < 0.05).

Ig: Immunoglobulins; SEM: standard error of the means; PL: p-Value of linear analysis; PQ: p-Value of quadratic analysis.

Table 8. Effects of B. coagulans on villi morphology of the small intestine in laying hens (n = 12).

Item	BC supplementation (g/t)					SEM	p-Value	PL	PQ
	0	50	100	150	200
Jejunum
Villus height, µm	1011.25^ab	1137.60^a	1013.49^ab	1061.56^ab	977.09^b	45.20	0.023	0.457	0.106
Crypt depth, µm	101.17	106.17	106.93	104.12	88.08	9.99	0.418	0.338	0.148
VH/CD	10.05	10.97	9.85	10.60	11.99	1.29	0.556	0.249	0.388
Ileum
Villus height, µm	683.45	629.31	762.47	771.36	695.64	53.99	0.085	0.090	0.201
Crypt depth, µm	115.4^ab	94.78^ab	86.54^b	104.24^ab	117.56^a	9.37	0.027	0.660	0.007
VH/CD	6.16^b	6.92^ab	8.85^a	7.44^ab	5.92^b	0.74	0.011	0.279	0.006

^a,bMeans sharing different letters in the same row are significantly different (p < 0.05).

VH/CD: villus height/crypt depth; BC: Bacillus coagulans; SEM: standard error of the means; PL: p-Value of linear analysis; PQ: p-Value of quadratic analysis.

Data availability statement

The results and analyses presented in this paper are freely available upon request.