Exogenous lysozyme influences Clostridium perfringens colonization and intestinal barrier function in broiler chickens

Figures & data

Table 1.  Composition of the diet and nutrient levels

Ingredient (g/kg)		Nutrient levels^a (g/kg)
Corn	280.7	Metabilizable energy (MJ/kg)	12.15
Wheat	330.0	Crude protein	210.0
Soybean meal (43%, C. perfringens)	321.6	Calcium	10.0
Soybean oil	25.7	Available phosphorus	4.5
Limestone	11.3	Lysine	11.5
Dicalcium phosphate	18.9	Methionine	5.0
l-Lysine (99%)	2.1	Methionine + cysteine	9.5
d,l-Methionine (98%)	1.8
Sodium chloride	3.5
Choline chloride (50%)	2.0
Mineral premix^b	2.0
Vitamin premix^c	0.2
Ethoxyquin (66%)	0.2
^aCalculated composition. ^bThe mineral premix supplied the following per kilogram of complete feed: copper, 8 mg; zinc, 75 mg; iron, 80 mg; manganese, 100 mg; selenium, 0.15 mg; iodine, 0.35 mg. ^cThe vitamin premix supplied the followings per kilogram of complete feed: vitamin A, 12,500 IU; vitamin D3, 2,500 IU; vitamin K3, 2.65 mg; vitamin B1, 2 mg; vitamin B2, 6 mg; vitamin B12, 0.025 mg; vitamin E, 30 IU; biotin, 0.0325 mg; folic acid, 1.25 mg; pantothenic acid, 12 mg; niacin, 50 mg.

Table 2.  Growth performance and intestinal lesion scores of broiler chickens

	Day 14	Days 14 to 28			Lesion score
Treatment	Initial body weight (g)	Weight gain (g)	Feed intake (g)	FCR	Day 22	Day 28
Control	291.0	680.8	1183.3	1.74	0.00	0.00^C
C. perfringens	300.7	616.9	1123.7	1.82	0.92	0.75^A
Lysozyme	293.4	689.5	1173.6	1.70	0.00	0.00^C
Lysozyme + C. perfringens	283.7	667.6	1148.5	1.72	0.67	0.50^B
SEM		9.565	15.406	0.016	0.090	0.073
P value
Lysozyme		0.082	0.812	0.017	0.078	0.037
C. perfringens		0.015	0.192	0.064	0.000	0.000
Lysozyme×C. perfringens		0.210	0.587	0.244	0.078	0.037
Means in the same column without the same uppercase superscript letter differ significantly (P <0.05). SEM, standard error of the mean.

Table 3.  C. perfringens, E. coli, Lactobacillus and Bifidobacterium. populations (log₁₀ colony-forming units/g) in the ileum of broiler chickens

	Day 22				Day 28
Treatment	C. perfringens	E. coli	Lactobacillus	Bifidobacterium	C. perfringens	E. coli	Lactobacillus	Bifidobacterium
Control	0.70	5.10	6.99	7.03	0.47^C	4.99	6.63	6.81
C. perfringens	4.79	7.01	7.68	7.18	3.60^A	6.38	7.44	6.94
Lysozyme	0.51	4.71	6.79	6.91	0.28^C	4.50	6.37	6.72
Lysozyme + C. perfringens	3.67	6.38	7.48	7.01	2.09^B	5.56	7.13	6.82
SEM	0.404	0.208	0.086	0.045	0.290	0.170	0.102	0.041
P value
Lysozyme	0.016	0.004	0.042	0.103	0.000	0.003	0.018	0.233
C. perfringens	0.000	0.000	0.000	0.183	0.000	0.000	0.000	0.173
Lysozyme×C. perfringens	0.074	0.435	0.993	0.797	0.000	0.395	0.854	0.833
Means in the same column with different uppercase superscript letter differ significantly (P <0.05). SEM, standard error of the mean.

Table 4.  Bacteria translocation rates of the liver and spleen (%) of broiler chickens

	Day 22				Day 28
	Liver		Spleen		Liver		Spleen
Treatment	Anaerobic bacteria	Aerobic bacteria	Anaerobic bacteria	Aerobic bacteria	Anaerobic bacteria	Aerobic bacteria	Anaerobic bacteria	Aerobic bacteria
Control	0.0(0/12)	0.0(0/12)^B	8.3(1/12)^B	8.3(1/12)^C	0.0(0/12)	0.0(0/12)	0.0(0/12)^C	0.0(0/12)^C
C. perfringens	33.3(4/12)	41.7(5/12)^A	100.0(12/12)^A	100.0(12/12)^A	16.7(2/12)	33.3(4/12)	75.0(9/12)^A	91.7(11/12)^A
Lysozyme	0.0(0/12)	0.0(0/12)^B	0.0(0/12)^B	8.3(1/12)^C	0.0(0/12)	0.0(0/12)	0.0(0/12)^C	8.3(1/12)^C
Lysozyme + C. perfringens	25.0(3/12)	33.3(4/12)^AB	75.0(9/12)^A	58.3(7/12)^B	16.7(2/12)	16.7(2/12)	25.0(3/12)^B	41.7(5/12)^BC
Means in the same column with different uppercase superscript letter differ significantly (P <0.05).

Table 5.  Lysozyme activity (mg/l) in the duodenum, jejunum and ileum of broiler chickens

	Day 22			Day 28
Treatment	Duodenum	Jejunum	Ileum	Duodenum	Jejunum	Ileum
Control	0.092	0.106	0.109	0.089^A	0.104	0.108
C. perfringens	0.027	0.023	0.119	0.043^C	0.051	0.127
Lysozyme	0.111	0.113	0.112	0.087^A	0.109	0.112
Lysozyme + C. perfringens	0.042	0.035	0.123	0.064^B	0.071	0.122
SEM	0.0082	0.0093	0.0043	0.0070	0.0060	0.0043
P value
Lysozyme	0.055	0.282	0.675	0.088	0.093	0.930
C. perfringens	0.000	0.000	0.243	0.000	0.000	0.093
Lysozyme×C. perfringens	0.785	0.750	0.933	0.041	0.344	0.621
Means in the same column with different uppercase superscript letter differ significantly (P <0.05). SEM, standard error of the mean.

Table 6.  Lysozyme g and g2 mRNA relative content in the duodenum of broiler chickens on day 22

Treatment	Lysozyme g/β-actin	Lysozyme g2/β-actin
Control	0.12	0.42
C. perfringens	0.13	0.43
Lysozyme	0.13	0.40
Lysozyme + C. perfringens	0.15	0.52
SEM	0.015	0.025
P value
Lysozyme	0.867	0.226
C. perfringens	0.197	0.519
Lysozyme×C. perfringens	0.289	0.263
SEM, standard error of the mean.