Effects of dietary supplementation of Trichoderma pseudokoningii fermented enzyme powder on growth performance, intestinal morphology, microflora and serum antioxidantive status in broiler chickens

Figures & data

Table 1. Ingredients and chemical composition of the basal diets of broilers.

Ingredient	Starter diet, g/kg	Finisher diet, g/kg
Yellow corn	472.6	518.0
Soybean meal (CP 44.0%)	345.2	295.9
Full fat soybean meal	100.0	100.0
Soybean oil	35.0	45.0
Calcium carbonate	16.2	13.4
Monocalcium phosphate	18.6	16.6
DL-Methionine	2.0	1.3
L-Lysine-HCl	3.7	3.2
NaCl	3.9	3.8
Choline-Cl	0.8	0.8
Vitamin premix^a	1.0	1.0
Mineral premix^b	1.0	1.0
Total	1000	1000
Calculated nutrient value
ME, kcal/kg	3050.1	3175.1
Crude protein, %	23.0	21.0
Calcium, %	1.05	0.90
Total phosphorus, %	0.75	0.70
Available phosphorus, %	0.50	0.45
Lysine, %	1.43	1.25
Methionine + cystein, %	1.07	0.96
Analyzed nutrition value
Crude protein, %	23.8	21.6
Dry matter, %	88.0	87.1

a Supplied per kg of diet: Vit A 15000 U; Vit. D3 3000 U; Vit. E 30 mg; Vit. K₃ 4 mg; Riboflavin 8 mg; Pyridoxine 5 mg; Vit. B₁₂ 25 μg; Ca-pantothenate 19 mg; niacin 50 mg; folic acid 1.5 mg; Biotin 60 μg.

b Supplied per kg of diet: Co (CoCO₃) 0.255 mg; Cu (CuSO4·5H₂O) 10.8 mg; Fe (FeSO4·H₂O) 90 mg; Zn (ZnO) 68.4 mg; Mn (MnSO4·H₂O) 90 mg; Se (Na₂SeO₃) 0.18 mg.

Table 2. The content of bioactive compounds and enzyme activities of Trichoderma pseudokoningii fermented enzyme powder^a.

	WB	EP
Total phenolic content,mg GAE^b/g DW^c	2.92 ± 0.05	4.79 ± 0.35
Xylanase, U/g	ND	487.96 ± 14.14
Cellulase, U/g	ND	18.55 ± 0.54

a The value is expressed as the mean ± standard deviation (n = 4).

b GAE: Gallic acid equivalent.

c DW: Dry weight.

WB: wheat bran; EP: enzyme powder; ND: not detected.

Figure 1. DPPH scavenging capacity (A), reducing power (B) and chelating capacity (C) of WB and EP aqueous extracts. Values are mean ± SD.

Table 3. Effect of Trichoderma pseudokoningii fermented enzyme powder supplemented in diet on growth performance of 1–35 d-old broilers.

Experimental period (d) and parameter	Experimental diets^1				SEM^3	p Values
	Control	0.1% EP	0.2% EP	0.4% EP
1–21 d
Body weight, g/bird^1	646.8^b	712.0^a	699.4^a	705.1^a	6.11	.009
Feed consumption, g/bird^2	805.0^b	873.7^a	874.3^a	867.7^a	5.60	.002
Weight gain, g/bird	603.1^b	668.5^a	655.9^a	661.7^a	6.09	.009
FCR	1.36^a	1.31^b	1.32^a^,^b	1.29^b	0.007	.050
22–35 d
Body weight, g/bird	1821.1	1932.7	1935.6	1948.3	29.96	.313
Feed consumption, g/bird	1816.8	1914.4	1896.4	1875.1	52.46	.616
Weight gain, g/bird	1174.3	1220.6	1236.2	1243.1	65.45	.796
FCR	1.63	1.59	1.55	1.51	0.014	.369
1–35 d
Feed consumption, g/bird	1310.9	1394.1	1385.3	1371.4	105.13	.953
Weight gain, g/bird	888.7	944.6	946.0	952.4	59.66	.975
FCR	1.48	1.45	1.43	1.40	0.027	.956

1 Results are provided as the means of four replicates (15 birds/replicate) in each control and treatment group (n = 4).

2 Results are provided as the means of 60 birds in each control and treatment group (n = 60).

3 SEM: standard error of the mean.

EP: enzyme powder.

a,b Means within the same rows without the same superscript letter are significantly different (p < .05).

Table 4. Effect of Trichoderma pseudokoningii fermented enzyme powder (EP) supplemented in diet on lactate concentration of intestinal content of 1–35 d-old broilers.

Lactate concentration^2, μM	Experimental diets^1				SEM^3	p Values
	Control	0.1% EP	0.2% EP	0.4% EP
Ileum
Day 21	64.3	68.7	64.0	67.2	1.84	.485
Day 35	44.0	43.8	47.5	45.3	1.24	.718
Overall	55.6	56.3	54.1	56.3	2.88	.991
Caecum
Day 21	110.8	113.9	119.8	118.9	2.62	.640
Day 35	95.1^c	113.4^a	115.4^a	105.6^b	6.52	.001
Overall	102.9^b	113.7^a^,^b	117.9^a	110.9^a^,^b	1.92	.013

1 Control: basal diet; 0.1% EP: 0.1% enzyme powder; 0.2% EP: 0.2% enzyme powder; 0.4% EP: 0.4% enzyme powder.

2 The results are provided as the means of four replicates (four birds/replicate) in each control and treatment group (n = 4). EP: enzyme powder.

3 SEM: standard error of the mean.

a–c Means within the same rows without the same superscript letter are significantly different (p < .05).

Table 5. Effect of Trichoderma pseudokoningii fermented enzyme powder (EP) supplemented in diet on microbial parameter in intestinal content of 1–35 d-old broilers.

	Experimental diets^1
Microbial parameter, log cfu/g	Control	0.1% EP	0.2% EP	0.4% EP	SEM^2	p Value
Day 21
Lactic acid bacteria
Ileum	8.15	8.05	8.08	8.42	0.10	.529
Caecum	10.79^b	11.17^a^,^b	11.88^a	11.29^a^,^b	0.11	.017
Coliform
Ileum	7.16	7.09	6.58	6.90	0.11	.334
Caecum	10.48	10.63	9.54	9.67	0.24	.079
Day 35
Lactic acid bacteria
Ileum	9.26	9.87	10.19	10.07	0.11	.071
Caecum	13.53	13.93	13.36	13.49	0.14	.602
Coliform
Ileum	9.67	9.06	8.62	8.89	0.22	.415
Caecum	11.57	10.37	9.79	9.94	0.25	.092
Overall
Lactic acid bacteria
Ileum	8.67	8.94	9.14	9.12	0.847	.200
Caecum	12.44	12.55	12.62	12.39	0.995	.340
Coliform
Ileum	8.41	8.08	7.81	7.90	0.873	.280
Caecum	11.01^a	10.20^a^,^b	9.46^b	9.43^b	0.013	.018

1 The results are provided as the means of four replicates (four birds/replicate) in each control and treatment group (n = 4).

2 SEM: standard error of the mean. EP: enzyme powder.

a,b Means within the same rows without the same superscript letter are significantly different (p < .05).

Figure 2. Photomicrography of jejunum and ileum of 21 and 35 d old broiler fed with control and fermented wheat brans. (A)–(D) Jejunum of 21 d broiler, control; 0.1% EP; 0.2%; 0.4% EP. (E)–(H) Jejunum of 35 d broiler, control; 0.1% EP; 0.2%; 0.4% EP. (I)–(L) Ileum of 21 d broiler, control; 0.1% EP; 0.2%; 0.4% EP. (M)–(P) Ileum of 35 d broiler, control; 0.1% EP; 0.2%; 0.4% EP haematoxylin and eosin stain. 40×.

Table 6. Effect of Trichoderma pseudokoningii fermented enzyme powder (EP) supplemented in diet on intestinal morphology of 1–35 d-old broilers.

Item	Experimental diets^1				SEM	p Values
	Control	0.1% EP	0.2% EP	0.4% EP
Jejunum
Day 21
Villus height, μm	1090.4^b	1140.6^a^,^b	1223.5^a	1215.7^a	16.48	.021
Crypt depth, μm	218.9	211.3	235.7	228.3	4.70	.270
Villus:crypt	4.85^b	5.49^a	5.31^a	5.39^a	0.09	.050
Day 35
Villus height, μm	1096.3^b	1331.2^a	1450.3^a	1317.6^a	24.30	<.0001
Crypt depth, μm	206.2	215.0	245.3	223.6	4.95	.084
Villus:crypt	5.43^b	6.20^a	6.17^a	5.99^a	0.10	.012
Ileum
Day 21
Villus height, μm	857.6	889.3	860.4	856.9	16.47	.282
Crypt depth, μm	147.5^b	155.2^a	142.4^a^,^b	141.6^a^,^b	2.43	.035
Villi:crypt	5.93^b	6.52^a	6.48^a	6.10^a	0.08	<.0001
Day 35
Villus height, μm	1166.4^a	1075.5^b	1081.2^b	1119.5^a^,^b	13.59	.039
Crypt depth, μm	214.8^a	173.5^b	180.1^b	185.8^b	3.76	.007
Villus:crypt	5.58^b	6.28^a	6.15^a	6.17^a	0.09	.016

1 The results are provided as the means of twenty spots corresponding to four birds for the control group and treatment groups, control: basal diet; 0.1% EP: 0.1% enzyme powder; 0.2% EP: 0.2% enzyme powder; 0.4% EP: 0.4% enzyme powder. EP: enzyme powder. SEM: standard error of the mean.

a,b Means within the same rows without the same superscript letter are significantly different (p < .05).

Table 7. Effects of Trichoderma pseudokoningii fermented enzyme powder (EP) supplemented in diets on the serum antioxidant enzymes activities and MDA levels of 1–35 d-old broilers.

Items	Experimental diets^1				SEM^2	p Values
	Control	0.1% EP	0.2% EP	0.4% EP
Day 21
SOD^3, U/mL	9.06	9.48	9.41	9.13	0.14	.071
CAT^3, U/mL	11.08^b	14.21^b	27.24^a	32.47^a	1.09	<.0001
MDA^3, μM	24.55	22.73	23.30	23.47	0.61	.761
Day 35
SOD, U/mL	8.54^c	9.03^b^,^c	9.39^b	10.26^a	0.10	<.0001
CAT, U/mL	13.48^b	15.84^a^,^b	24.20^a	19.71^a^,^b	1.23	.008
MDA, μM	28.01	27.61	25.91	29.12	0.70	.455
Overall
SOD, U/mL	8.86^c	9.22^b^,^c	9.40^a^,^b	9.83^a	0.08	.001
CAT, U/mL	11.73^b	19.05^a^,^b	25.42^a	23.27^a	1.48	.027
MDA, μM	26.28	25.17	24.60	26.11	0.53	.637

1 Control: basal diet; 0.1% EP: 0.1% enzyme powder; 0.2% EP: 0.2% enzyme powder; 0.4% EP: 0.4% enzyme powder. Results are given as the means of 16 birds’ serum samples for each treatment at day 21 and 35.

2 SEM: standard error of the mean.

3 CAT, SOD and MDA represent catalase, superoxide dismutase and malondialdehyde, respectively.

a–c Means within the same rows without the same superscript letter are significantly different (p < .05).

Table 8. Evaluation of the economic benefit of Trichoderma pseudokoningii-fermented enzyme powder (EP) supplemented in diet.

Item	Experimental diets
	Control	0.1% EP	0.2% EP	0.4% EP
Feed cost, NT$/bird
1–35 d	26.3	28.3	28.7	29.6
Meat income, NT$/bird
1–35 d	81.9	87.0	87.1	87.7
Income over feed cost, NT$/bird
1–35 d	55.6	58.7	58.4	58.1

Feed cost: based on the costs (NT$/kg) of the ingredients as follows: yellow corn 10.48, soybean meal 13.63, full fat soybean meal 18.3, soybean oil 38.00, monocalcium phosphate 16.95, calcium carbonate 1.94, l-Lysine-HCl 300, dl-Methionine 230, salt (NaCl) 2.70, choline chloride, 50% 33.8, vitamin premix 109.5, and mineral premix 43.0. The fees for processing of basal ration per kg were 9.00 for grain mixture of control group, 10.10 for 0.1% EP group, 10.03 for 0.2% EP group and 10.36 for 0.4% EP group during 1–21 d, respectively. The fees for processing of basal ration per kg were 17.33 for grain mixture of control group, 18.24 for 0.1% EP group, 18.70 for 0.2% EP group and 19.26 for 0.4% EP group during 22–35 d, respectively.