Effects of different levels of Hermetia illucens larvae meal on performance, egg quality, yolk fatty acid composition and oxidative status of laying hens

Figures & data

Table 1. Composition and nutrient levels of diets (air-dry basis).

Items	CON	HILM-1	HILM-3	HILM-5
Ingredients^a (%, unless otherwise indicated)
Corn	63.80	62.10	58.00	54.00
Soybean meal	23.20	22.00	19.50	17.00
Wheat bran	0.00	1.90	6.50	11.00
Limestone	7.00	7.00	7.00	7.00
HILM^b	0.00	1.00	3.00	5.00
Ca(HCO3)2	1.00	1.00	1.00	1.00
Premix^c	5.00	5.00	5.00	5.00
Total	100.00	100.00	100.00	100.00
Calculated composition
ME (MJ/kg)	10.97	11.00	11.00	11.02
CP	15.81	15.80	15.81	15.81
EE	2.74	3.09	3.80	4.51
Ca	2.89	2.91	2.95	2.99
Lys	0.89	0.90	0.90	0.91
AP	0.54	0.56	0.58	0.61
Met	0.45	0.48	0.50	0.52

^aHILM, Hermetia illucens larvae meal; ME, metabolisable energy; CP, curde protein; EE, ether extract; Met, methionine; Lys, lysine; Ca, Calcium; AP, available phosphorus.

^bCON, HILM-1, HILM-2, HILM-3 diets contained 0, 1%, 3%, 5% HILM, respectively.

^cThe premix provided the following per kg of diets：vitamin A, 14 000 IU; vitamin B, 13 mg; vitamin B₂, 12 mg; vitamin B₅, 16.5 mg; vitamin B₆, 6 mg; vitamin B₁₂, 0.03 mg; vitamin D₃, 3 500 IU; vitamin E, 100 IU; vitamin K₃, 4.25 mg; nicotinamide, 50 mg; D-biotin, 0.25 mg; folic acid, 2 mg; Cu, 12.5 mg; Fe, 70 mg; Mn, 110 mg; Zn, 85 mg; I, 1.25 mg; Se, 0.3 mg; Ca, 5 g; TP, 1.5 g; NaCl, 1.5 ∼ 7.5 g; choline chloride, 400 mg.

Table 2. Effects of HILM on production performance of laying hens^A.

Item^B	Dietary treatment^C				Pooled SEM	p-Value
	CON	HILM-1	HILM-3	HILM-5		ANOVA	Linear	Quadratic
Day 1-28 of the experiment
Egg weight (g)	48.65^b	48.36^b	49.58^a	48.64^b	0.128	.003	.281	.182
Feed intake (g/d per hen)	85.37^b	88.66^ab	91.89^a	89.71^a	0.596	.001	.002	.016
Egg production (%)	54.27	54.50	56.95	57.56	0.007	.172	.035	.887
FCR (g of feed/g of egg)	3.29	3.41	3.27	3.29	0.039	.534	.672	.481
Day 29-56 of the experiment
Egg weight (g)	49.04^b	49.04^b	50.07^a	49.30^b	0.125	.009	.100	.113
Feed intake (g/d per hen)	85.14	88.18	90.05	87.55	0.750	.139	.172	.065
Egg production (%)	49.95^b	47.18^b	50.03^b	55.32^a	0.007	.001	.003	.005
FCR (g of feed/g of egg)	3.51^ab	3.75^a	3.59^ab	3.26^b	0.049	.003	.027	.003
Day 1-56 of the experiment
Egg weight (g)	48.85^b	48.70^b	49.83^a	48.97^b	0.091	.000	.059	.042
Feed intake (g/d per hen)	85.26^b	88.42^ab	90.97^a	88.63^ab	0.480	.000	.002	.003
Egg production (%)	52.11^ab	50.84^b	53.49^ab	56.44^a	0.524	.001	.001	.039
FCR (g of feed/g of egg)	3.40^a	3.57^a	3.43^ab	3.27^b	0.032	.010	.059	.009

^AData are means of six replicates per treatment with 18 hens per replicate.

^BHILM, Hermetia illucens larvae meal; FCR, feed conversion ratio.

^CCON, HILM-1, HILM-2, HILM-3 diets contained 0, 1%, 3%, 5% HILM, respectively.

^a-bMeans within a row with different superscripts differ significantly (p < .05).

Table 3. Effect of HILM on egg quality of laying hens^A.

Item^B	Dietary treatment^C				Pooled SEM	p-Value
	CON	HILM-1	HILM-3	HILM-5		ANOVA	Linear	Quadratic
Day 28 of the experiment
Egg-shape index	1.30	1.30	1.30	1.33	0.006	.273	.108	.267
Eggshell-breaking strength (kgf)	3.90	3.74	3.94	3.88	0.060	.663	.780	.668
Eggshell thickness (mm)	0.35^a	0.35^a	0.32^b	0.33^ab	0.004	.025	.026	.626
Yolk index	0.41^a	0.40^a	0.39^ab	0.37^b	0.004	.001	.000	.437
Haugh unit	62.10^b	63.84^ab	66.32^a	64.42^ab	0.574	.046	.065	.110
Albumen height (mm)	4.33^a	4.15^ab	4.15^ab	3.64^b	0.073	.005	.001	.241
Eggshell weight (g)	4.71^b	4.47^b	5.38^a	4.59^b	0.056	.000	.179	.004
Day 56 of the experiment
Egg-shape index	1.36	1.32	1.33	1.34	0.009	.550	.468	.243
Eggshell-breaking strength (kgf)	4.18	3.64	3.83	3.47	0.128	.234	.090	.745
Eggshell thickness (mm)	0.37	0.34	0.36	0.32	0.007	.101	.087	.815
Yolk index	0.40	0.42	0.38	0.40	0.006	.175	.341	1.000
Haugh unit	64.51	57.89	65.55	62.73	1.186	.107	.821	.412
Albumen height (mm)	4.62	4.59	5.12	4.60	0.139	.493	.700	.392
Eggshell weight (g)	4.71	4.33	4.40	4.35	0.065	.143	.085	.206

^AData are means of six replicates per treatment with 18 hens per replicate.

^BHILM, Hermetia illucens larvae meal.

^CCON, HILM-1, HILM-2, HILM-3 diets contained 0, 1%, 3%, 5% HILM, respectively.

^a-bMeans within a row with different superscripts differ significantly (p < .05).

Table 4. Effect of HILM on yolk fatty acid composition of laying hens^A.

Item^B	Dietary treatment^C				Pooled SEM	p-Value
	CON	HILM-1	HILM-3	HILM-5		ANOVA	Linear	Quadratic
C14:0	0.45^b	0.53^b	0.69^a	0.84^a	0.039	.000	.000	.375
C16:0	26.68	26.76	25.88	25.52	0.230	.149	.036	.612
C17:0	0.16^b	0.15^b	0.20^a	0.20^a	0.007	.018	.006	.783
C18:0	8.75	9.18	9.03	9.02	0.103	.566	.504	.317
SFA	36.05	36.62	35.79	35.58	0.236	.469	.311	.424
C14:1	0.15	0.15	0.18	0.19	0.009	.275	.071	.911
C16:1	4.09	3.72	3.48	3.10	0.141	.075	.011	.984
C18:1n9t	0.18	0.19	0.18	0.18	0.004	.559	.408	.584
C18:1n9c	42.32	42.06	41.62	42.18	0.391	.941	.822	.633
C20:1	0.25	0.24	0.26	0.25	0.006	.855	.567	1.000
MUFA	46.97	46.34	45.71	45.89	0.330	.573	.220	.559
C18:2n6c	11.98	12.14	13.46	13.60	0.309	.118	.026	.986
C18:3n3	0.38	0.37	0.40	0.34	0.013	.338	.478	.241
C18:3n6	0.16	0.16	0.19	0.15	0.005	.140	.742	.086
C20:2	0.15^b	0.18^ab	0.20^a	0.16^ab	0.006	.046	.242	.013
C20:3n6	0.27	0.28	0.28	0.24	0.009	.529	.393	.241
C20:4n6	2.90	2.92	2.88	2.96	0.041	.910	.713	.705
C22:6n3	1.10	1.04	1.12	1.07	0.022	.634	.939	.863
PUFA	16.94	17.04	18.46	18.51	0.312	.114	.028	.965

^AData are means of six replicates per treatment with 2 hens per replicate;

^BHILM, Hermetia illucens larvae meal; SFA, saturated fatty acids; MUFA, monounsaturated fatty acids; PUFA, polyunsaturated fatty acids;

^CCON, HILM-1, HILM-3, HILM-5 diets contained 0, 1%, 3%, 5% HILM, respectively;

^a-bMeans within a row with different superscripts differ significantly (p < .05).

Table 5. Effect of HILM on yolk amino acid composition of laying hens^A (mg/g).

Item^B	Dietary treatment^C				Pooled SEM	p-Value
	CON	HILM-1	HILM-3	HILM-5		ANOVA	Linear	Quadratic
NEAA
Aspartic Acid (Asp)	46.79	48.16	48.44	47.96	0.545	.752	.469	.428
Glutamic Acid (Glu)	56.93^b	58.00^ab	61.40^a	60.23^ab	0.626	.024	.009	.264
Arginine (Arg)	27.16	28.18	28.49	27.56	0.330	.508	.619	.163
Proline (Pro)	16.89	17.30	17.73	17.20	0.203	.565	.479	.270
Alanine (Ala)	23.95	24.76	25.19	24.61	0.295	.548	.382	.266
Serine (Ser)	33.25	34.03	34.95	33.59	0.388	.472	.585	.192
Glycine (Gly)	14.14	14.56	15.03	14.53	0.186	.433	.342	.230
EAA
Histidine (His)	10.33	10.72	10.85	10.43	0.142	.562	.756	.180
Threonine (Thr)	20.83	21.51	21.98	21.33	0.245	.449	.388	.195
Tyrosine (Tyr)	14.12	14.65	15.53	14.81	0.216	.136	.115	.138
Cystine (Cys)	4.05	4.11	4.45	4.39	0.079	.181	.056	.685
Valine (Val)	26.60^b	27.07^ab	29.17^a	27.82^ab	0.352	.046	.042	.141
Methionine (Met)	7.58^b	8.65^b	9.96^a	8.41^b	0.248	.001	.013	.001
Phenylalanine (Phe)	22.82^b	22.77^b	25.05^a	23.46^b	0.323	.039	.088	.172
Leucine (Leu)	37.50^b	37.57^b	40.62^a	38.48^ab	0.452	.048	.087	.170
Isoleucine (Ile)	22.60	23.37	23.89	23.11	0.310	.557	.477	.242
Lysine (Lys)	29.50	30.91	30.94	30.49	0.361	.492	.374	.223
Total	416.31	430.26	440.35	427.32	5.177	.464	.369	.215

^AData are means of six replicates per treatment with 2 hens per replicate;

^BHILM, Hermetia illucens larvae meal; NEAA, nonessential amino acid; EAA, essential amino acid;

^CCON, HILM-1, HILM-3, HILM-5 diets contained 0, 1%, 3%, 5% HILM, respectively;

^a-b Means within a row with different superscripts differ significantly (p < .05).

Table 6. Effect of HILM on plasma antioxidant capacity of laying hens^A.

Item^B	Dietary treatment^C				Pooled SEM	p-Value
	CON	HILM-1	HILM-3	HILM-5		ANOVA	Linear	Quadratic
GSH-PX (U/mL)	7005.02	7096.83	6037.35	6621.26	230.609	.387	.298	.600
T-SOD (U/mL)	150.46^b	202.88^a	211.82^a	181.23^ab	7.888	.017	.102	.005
T-AOC (U/mL)	4.46	4.48	5.16	4.15	0.292	.690	.928	.405
MDA (nmol/mL)	7.86^ab	8.93^a	4.70^ab	4.17^b	0.667	.024	.006	.486
CAT (U/mL)	3.76	4.52	5.98	5.87	0.431	.197	.047	.605

^AData are means of two hens of six replicates per treatment.

^BHILM, Hermetia illucens larvae meal; GSH-Px, glutathione peroxidase; T-SOD, total superoxide dismutase; T-AOC, total antioxidant capacity; MDA, malondialdehyde; CAT, catalase.

^CCON, HILM-1, HILM-3, HILM-5 diets contained 0, 1%, 3%, 5% HILM, respectively.

^a-bMeans within a row with different superscripts differ significantly (p <.05).

Table 7. Effects of HILM on plasma immune indexes of laying hens^A.

Item^B	Dietary treatment^C				Pooled SEM	p-Value
	CON	HILM-1	HILM-2	HILM-3		ANOVA	Linear	Quadratic
IgG (g/L)	4.53	4.56	4.41	4.53	0.035	.461	.586	.523
IgM (g/L)	1.74	1.75	1.68	1.72	0.016	.428	.330	.651
IgA (g/L)	2.32	2.35	2.23	2.23	0.030	.398	.149	.802
NDV-Ab (S/P)	0.75	0.81	0.90	0.95	0.033	.096	.014	.967
AIV (S/P)	0.99^b	0.99^ab	1.07^a	1.01^ab	0.010	.044	.149	.116
IFN-γ (pg/mL)	44.97	47.77	37.07	44.53	2.058	.353	.504	.575
IL-2 (pg/mL)	249.96^ab	265.03^a	178.17^b	202.08^ab	12.125	.028	.017	.827
IL-4 (pg/mL)	14.26	12.54	11.75	12.33	0.755	.696	.338	.480

^AData are means of two hens of six replicates per treatment.

^BHILM, Hermetia illucens larvae meal; IgG, immunoglobulin G; IgM, immunoglobulin M; IgA, immunoglobulin A; NDV-Ab, newcastle disease virus antibody; AIV-Ab, avian influenza virus antibody; IFN-γ, interferon-γ; IL-2, interleukin- 2; IL-4, interleukin- 4.

^CCON, HILM-1, HILM-3, HILM-5 diets contained 0, 1%, 3%, 5% HILM, respectively.

^a-bMeans within a row with different superscripts differ significantly (p < .05).