Impact of probiotics and/or organic acids supplementation on growth performance, microbiota, antioxidative status, and immune response of broilers

Figures & data

Table 1. Ingredients and composition of the starter-grower and finisher diets (as-fed basis).

Item (%)	Starter-grower	Finisher
	(1–28 d)	(28–42 d)
Yellow corn	53.38	60.18
Soybean meal (44% crude protein)	31.8	25.6
Gluten (60% crude protein)	7.7	6.2
Vegetable oil	3	4.3
Dicalcum phosphate^a	1.6	1.2
Limestone	1.4	1.4
Salt	0.4	0.4
DL-methionine	0.07	0.07
Lysine-HCL	0.15	0.15
Premix^b	0.5	0.5
Total	100	100
Calculated composition
Crude protein (%)	23.03	20.05
Metabolizable energy (kcal /kg)	3,050	3,194
Calcium (%)	1.01	0.90
Available phosphorous (%)	0.41	0.33
Ether extract (%)	7.48	8.49

^aDicalcium phosphate contained 16% phosphorous and 23% calcium.

^bPremix provided per kilogram of diet: vitamin A (transretinyl acetate), 10,000 U; vitamin D₃ (cholecalciferol), 3,000 U; vitamin E (all-rac-α-tocopherol), 30 U; menadione, 1.3 mg; thiamine, 2.2 mg; riboflavin, 8 mg; nicotinamide, 40 mg; choline chloride, 400 mg; calcium pantothenate, 10 mg; pyridoxine, 4 mg; biotin, 0.04 mg; folic acid, 1 mg; vitamin B₁₂ (cobalamin), 0.013 mg; Fe (from ferrous sulphate), 80 mg; Cu (from copper sulphate), 8.0 mg; Mn (from manganese sulphate), 110 mg; Zn (from zinc oxide), 60 mg; I (from calcium iodate), 1.1 mg; Se (from sodium selenite), 0.3 mg.

Table 2. Effect of probiotic and organic acids supplementation on growth performance and mortality rate of broiler chickens.

Item	Probiotic, P, %		Organic acids, OA, %			SEM	p-Value
	0	0.02	0	0.03	0.06		P	OA	P × OA
Body weight at 6 wk, g	2417.8^b	2458.7^a	2413.2	2448.0	2454.0	14.5	.01	.07	.57
Feed intake, g/bird	4421.8	4400.8	4373.5	4423.4	4437.1	28.92	.73	.66	.77
Feed conversion ratio, g feed/g gain	1.89	1.84	1.86	1.87	1.86	0.02	.14	.97	.89
Mortality, %	2.83	2.33	3.50	1.50	2.80	0.48	.61	.24	.32

^a,bMeans within a row with different superscript letters for each factor are significantly different.

Abbreviations: p-Value, Probability; SEM, standard error of mean.

Table 3. Effect of probiotic and organic acids supplementation on carcase traits of broiler chickens.

Item	Probiotic, P, %		Organic acids, OA, %			SEM	p-Value
	0	0.02	0	0.03	0.06		P	OA	P × OA
Carcase, %	71.20	72.36	71.88	71.77	71.70	0.32	.07	.97	.34
Whole leg, %	13.56	13.37	13.28	13.40	13.72	0.16	.59	.55	.89
Thigh, %	8.23	8.50	8.16	8.37	8.56	0.14	.33	.52	.36
Drumstick, %	4.57	4.53	4.56	4.44	4.65	0.05	.63	.26	.70
Major pectorals, %	9.99	10.22	10.47	9.71	10.14	0.16	.48	.16	.83
Minor pectorals, %	1.99	2.03	2.04	2.00	1.98	0.03	.56	.80	.76
Liver, %	1.81	1.85	1.84	1.86	1.80	0.02	.38	.60	.88
Heart, %	0.39	0.40	0.40	0.39	0.40	0.005	.08	.91	.47
Gizzard, %	1.48	1.42	1.47	1.44	1.44	0.03	.29	.91	.15
Bursa of Fabricius, %	0.05	0.05	0.05	0.05	0.05	0.002	.86	.45	.28
Spleen, %	0.11	0.12	0.11	0.11	0.12	0.003	.29	.39	.30

^a,bMeans within a row with different superscript letters for each factor are significantly different.

Abbreviations: p-Value, Probability; SEM, standard error of mean.

Figure 1. Interaction between probiotics (0% and 0.2%) and organic acids (0, 0.03, and 0.06%) supplementation on plasma concentrations of glutathione peroxidase (GSH-Px) and globulin, and cell-mediated immune response at 24 h post-phytohaemagglutinin-P injection (toe web swelling difference). Different superscripts in a column differ significantly (p < .05). Abbreviations: P, probiotics; OA, organic acids.

Table 4. Effect of probiotic and organic acids supplementation on blood biochemical constituents and antioxidative status of broiler chickens.

Item	Probiotic, P, %		Organic acids, OA, %			SEM	p-Value
	0	0.02	0	0.03	0.06		P	OA	P × OA
Total Protein, g/dL	3.33^b	3.55^a	3.35^b	3.56^a	3.41^ab	0.04	.002	.04	.18
Albumin, g/dL	1.53^b	1.70^a	1.61	1.63	1.61	0.03	.007	.95	.09
Globulin, g/dL	1.80	1.85	1.74^b	1.93^a	1.80^ab	0.03	.40	.04	.003
Cholesterol, mg/dL	111.87	108.40	116.85	108.10	105.45	5.98	.48	.14	.66
Triglycerides, mg/dL	74.50	67.13	73.75	71.00	67.70	7.42	.16	.64	.30
T3, ng/mL	2.72	3.11	2.64	3.12	2.95	0.24	.10	.26	.58
T4, ng/mL	3.17	4.06	3.38	4.33	3.59	0.48	.33	.79	.66
GSH-Px, U/mL	38.47^b	47.78^a	43.77	42.80	42.80	1.66	.003	.95	.02
TAC, mmol/L	0.84	0.96	0.82	0.92	0.96	0.08	.26	.51	.45
MDA, nmol/mL	6.67	6.36	6.37	6.45	6.77	0.26	.53	.82	.62

^a,bMeans within a row with different superscript letters for each factor are significantly different.

Abbreviations: p-Value, Probability; SEM, standard error of mean; T₃, triiodothyronine; T₄, thyroxine; GSH-Px, glutathione peroxidase; TAC, total antioxidant capacity; MDA, malondialdehyde.

Table 5. Effect of probiotic and organic acids supplementation on cellular and humoral immune response of broiler chickens.

Item	Probiotic, P, %		Organic acids, OA, %			SEM	p-Value
	0	0.02	0	0.03	0.06		P	OA	P × OA
Toe web swelling difference (mm)
24 h	0.53^b	0.65^a	0.50^b	0.64^a	0.63^a	0.22	.01	.01	.03
48 h	0.35^b	0.42^a	0.34^c	0.43^a	0.39^b	0.01	.04	.05	.26
72 h	0.26	0.33	0.26	0.34	0.28	0.02	.09	.21	.60
Immunoglobulin (mg/ml)
IgY	5.16^b	8.66^a	5.84	7.24	7.65	1.93	.03	.62	.69
IgM	1.60	1.74	1.62	1.66	1.72	0.07	.33	.85	.27
IgA	0.20	0.26	0.21	0.23	0.26	0.03	.18	.70	.94
NDV titre	4515.2^b	5466.3^a	5059.3	5343.7	4565.8	595.3	.01	.21	.23

^a,b,cMeans within a row with different superscript letters for each factor are significantly different.

Abbreviations: p-Value, Probability; SEM, standard error of mean; IgY, Immunoglobulin-Y; IgM, Immunoglobulin-M; IgA, Immunoglobulin-A; NDV, Newcastle Disease Virus.

Figure 2. Interaction between probiotics (0% and 0.2%) and organic acids (0, 0.03, and 0.06%) supplementation on bacterial count in meat and gut. Different superscripts in a column differ significantly (p < .05). Abbreviations: P, probiotics; OA, organic acids.

Table 6. Effect of probiotic and organic acids supplementation on bacterial count in gut and breast meat of broiler chickens.

Item	Probiotic (P, %)		Organic acids (OA, %)			SEM	p-Value
	0	0.02	0	0.03	0.06		P	OA	P × OA
Gut bacterial count
Total count (log10 cfu/g)	7.03^a	5.12^b	6.31	6.32	5.60	0.28	.001	.19	.47
B. subtilis (log10 cfu/g)	0.00^b	4.51^a	2.16	2.31	2.31	0.54	.001	.81	.81
E. coli (log10 cfu/g)	5.04	4.74	5.69^a	5.05^a	3.93^b	0.20	.23	.001	.55
Salmonella (%)	66.67^a	33.33^b	80.00^a	40.00^b	30.00^c	6.51	.001	.001	.001
Breast-meat bacterial count
Total count (log10 cfu/g)	6.21^a	4.36^b	6.32^a	5.67^a	3.88^b	0.39	.001	.001	.01
B. subtilis (log10 cfu/g)	0.00^b	3.52^a	1.69^b	1.86^a	1.71^b	0.42	.001	.01	.01
E. coli (log10 cfu/g)	3.86	3.13	4.37 ^a	3.35 ^b	2.75 ^b	0.28	.18	.05	.75
Salmonella (%)	Negative	Negative	Negative	Negative	Negative	–	–	–	–

^a,b,cMeans within the same column with different superscript are significantly differed.

Abbreviations: p-Value, Probability; SEM, standard error of mean.

Data availability statement

Data and materials that support the results and analyses presented in this study are freely available upon request.