The effect of Artemisia annua on broiler performance, on intestinal microbiota and on the course of a Clostridium perfringens infection applying a necrotic enteritis disease model

Figures & data

Table 1.  Ingredients and calculated composition of the basic diet.

Ingredient	Basic feed (g/kg as fed)
Wheat	499.1
Maize	100.0
Rape seed (double low)	40.0
Soya bean meal (de-hulled))	294.1
Soya bean oil	24.5
Calcium carbonate	9.0
Monocalcium phosphate	15.6
Methionine 100%	3.5
Threonine 98%	1.0
Lysine 100%	2.5
Sodium chloride	2.0
Sodium bicarbonate	2.7
Vitamin/mineral premix^a	6.0
Calculated
Energy (MJ/kg)	12.39
Protein	220.02
Fat	61.64
Crude fibre	26.52
Ash	56.00
Water	125.84
Starch	355.56
Sugar	46.91
Lysine	12.98
Methionine	6.43
Cystine	3.60
Threonine	8.74
Calcium	8.99
Phosphorus, available	4.84
Phosphorus total	7.25
Sodium	1.60
Potassium	8.91
Chloride	1.87
^aSupplying per kilogram of diet: retinol, 12,000 IU; cholecalciferol, 5000 IU; tocopherol, 62 IU; menadione, 5 mg; thiamin, 2 mg; riboflavin, 8 mg; pyridoxine, 4 mg; d-pantothenic acid; 12 mg; niacin, 60 mg; folic acid, 3 mg; biotin, 0.2 mg; cyanocobalamin, 0.02 mg; betaine anhydrate, 260 mg; calcium, 0.22 g; sodium, 1.35 g; chloride, 1.12 g; potassium, 3 mg; iron, 20 mg; zinc, 100 mg; manganese, 100 mg; copper, 26 mg; iodine, 2 mg; selenium, 0.2 mg.

Table 2.  Chemical composition of dried ground A. annua leaves.

Item	Sample 1	Sample 2
Dry matter (g/kg)	891	896
Ash (g/kg dry matter)	124	120
Fat (g/kg dry matter)	43	44
Crude protein (g/kg dry matter)	30	27
Starch (g/kg dry matter)	6	6
Insoluble NSP^a (g/kg dry matter)	162	163
Soluble NSP (g/kg dry matter)	93	76
Klason lignin (g/kg dry matter)	227	191
Dietary fibre^b (g/kg dry matter)	482	430
^aNSP, non-starch polysaccharides.
^bDietary fibre: total NSP + lignin.

Table 3.  Production results of broilers receiving increasing dietary concentrations of dried and ground A. annua leaves (Experiment 1) or n-hexane extract (Experiment 2).

	Dried plant leaves of A. annua (g/kg)
Experiment 1	0	5	10	20	SEM	P value
1 to 35 days
Body weight	2041^A	2040^A	1938^B	1876^B	33	0.013
Feed intake	2902^A	2953^A	2580^B	2543^B	91	0.009
FCR (kg feed/kg gain)	1.42	1.45	1.33	1.35	0.03	0.099
	n-Hexane extract of A. annua (mg/kg)
Experiment 2	0	125	250	500	SEM	P value
1 to 35 days
Body weight	2078	2021	2060	1951	35	0.105
Feed intake	3088^A	2948^A	2846^B	2644^B	78	0.011
FCR (kg feed/kg gain)	1.49	1.46	1.38	1.36	0.03	0.069
Values are least-square means (n=4). Means in the same row with different uppercase superscript letters differ significantly (P<0.05). FCR, feed conversion ratio; SEM, standard error of the mean.

Table 4.  Numbers of selected bacteria in the ileum and caeca contents (log colony-forming units/g digesta) of C. perfringens challenged broilers on day 27.

	Control non-challenged	Control challenged	Dried plant 10 g/kg challenged	n-Hexane extract 250 mg/kg challenged	SEM	P value
Ileum
Lactic acid bacteria	8.69	8.67	8.73	8.71	0.143	0.995
Coliform bacteria	5.31^B	7.88^A	8.04^A	7.96^A	0.550	0.011
Lactose-negative enterobacteria	5.85	6.44	6.27	6.63	0.523	0.751
Anaerobic bacteria	8.86	8.70	8.72	8.74	0.119	0.785
Caeca
Lactic acid bacteria	8.64^C	9.36^A	9.12^AB	8.79^BC	0.153	0.025
Coliform bacteria	8.12	7.85	6.91	6.66	0.659	0.368
Lactose-negative enterobacteria	6.38	6.33	5.86	5.42	0.516	0.539
Anaerobic bacteria	9.89	10.03	9.98	9.62	0.158	0.301
Values are least-square means (n=4). Means in the same row with different uppercase superscript letters differ significantly (P<0.05). SEM, standard error of the mean.

Table 5.  Small intestinal lesion scores and numbers of C. perfringens in ileum and caeca of broilers following challenge with C. perfringens strain 200302-1-1-Ba.

Item	Control non-challenged	Control challenged	Dried plant 10 g/kg challenged	n-Hexane extract 250 mg/kg challenged	SEM		P value
							Diet	Day	Diet×day
Lesion score							0.05	0.0001	0.81
22 days	0^a	1.75 (17)	1.30 (17)	0.60 (9)	0.36	Y^b
24 days	0	3.65 (20)	2.75 (17)	2.65 (18)	0.36	X
27 days	0	1.75 (17)	1.30 (18)	0.60 (9)	0.36	Y
Difference^c		A	A	B
Clostridium perfringens, ileum							0.0001	0.0001	0.66
22 days	3.16	8.07	7.52	7.20	0.27	Y
24 days	3.98	8.42	8.44	7.93	0.29	X
27 days	2.40	6.37	6.31	6.31	0.29	Z
Difference^c	C	A	AB	B
Clostridium perfringens, caeca							0.0001	0.0001	0.25
22 days	3.27	8.31	7.92	7.10	0.34	Y
24 days	5.45	8.53	8.30	7.90	0.55	X
27 days	2.31	6.83	7.23	6.09	0.20	Z
Difference^c	C	A	A	B
Values are least-square means (n=4) (number of birds with lesions out of 20 birds).
^aControl, non-challenged group not included in the statistical analysis.
^bDays within an item with different letters (X, Y, Z) differ significantly (P≤0.05).
^cDietary groups within an item with different uppercase letters (A, B, C) differ significantly (P≤0.05).

Figure 1.  Weight gain of broilers during the period post infection from 17 to 27 days.

Values are least square means (n=4), SEM=14.2, P diet<0.001.

Bars with different letters differ significantly (P≤0.05).