Immune function, egg quality and production responses in layer hens fed two different lignocellulose fibre supplements during the early laying period

Figures & data

Table 1. Chemical composition (g/100 g) of commercial diets, ‘Pullet grower’ and ‘Golden yolk’, used as the Control and basal experimental diets (manufacturer’s information^a).

Chemical Composition^b	Pullet Grower	Golden Yolk
Dry matter	86.0		86.0
Moisture	14.0		14.0
Protein	15.5		15
Fat	2.5		2.5
Fibre (maximum)^c	8.0		10.0
ME kcal/kg	2700		2800
Calcium	1.0		3.8
Phosphorus	0.64		0.68
Available Phosphorus	0.5		0.35

^aRidley AgriProducts Pty Ltd., Pakenham, 3810, Australia.

^bFeed analysis and ingredients were obtained from manufacture but for confidentiality reasons the quantities used were not able to be obtained.

Ingredients: wheat, barley, field peas, meat and bone meal, beef tallow, canola meal expeller, soybean meal, canola seed, Millrun mix, oat hulls, limestone, MDCP, salt, sodium bicarb, choline chloride, methionine, lysine, vitamin and mineral premix, phytase and xylanase.

^cCrude fibre Analysed by FeedTest (Agrifood Technology Pty Ltd, Werribee, Vic., Australia) in Golden Yolk was 6.4%.

ME: metabolisable energy.

Table 2. Live body weights (BW), weights and relative weights (RW, g/100 g BW) of immune organs, Peyer’s patches (PP) measurements and jejunal plus ileal areas of Hy-Line Brown layer strain hens given diets containing different types of lignocellulose fibres for 8 weeks from 20 weeks of age (mean, pooled SEM, n = 6 pens).

Treatment	C	LC1	LC2	p Value	Pooled SEM
Initial BW (g)	1729.3	1722.8	1726.3	0.973	10.6
Final BW (g)	1939.5	1992.8	1956.9	0.192	12.1
Absolute organ weights (g)
Bursa of Fabricius	0.380	0.397	0.394	0.906	0.02
Thymus glands	0.360^a	0.485^b	0.427^ab	0.020	0.02
Spleen	1.96	2.10	1.91	0.202	0.04
Caecal tonsils	0.208	0.245	0.220	0.417	0.01
Organ RW (g/ 100 g BW)
Bursa of Fabricius	0.0196	0.0200	0.0202	0.965	0.001
Thymus glands	0.0187^a	0.0244^b	0.0217^ab	0.043	0.001
Spleen	0.1012	0.1046	0.0974	0.391	0.002
Caecal tonsils	0.0107	0.0123	0.0113	0.581	0.001
Number of PP	8.8^a	12.7^b	9.2^a	0.001	0.5
Area of PP (cm^2)	3.1^a	4.2^b	3.5^ab	0.007	0.2
Jejunum + ileum area (cm^2)	187.1	201.5	184.5	0.221	4.2
Relative area of PP (% of area of jejunum + ileum)	1.7^a	2.1^b	1.9^ab	0.022	0.1

^a,bMeans within the same row with different subscript letters are significantly different (p < 0.05).

To calculate a mean for each pen, three hens/pen were used for measurements of immune organ weights and 1 hen/pen for PP measurements and jejunal and ileal areas.

^C Control diet – see Table 1. LC1 - Control diet plus 1.5 g/100 g Arbocel RC fine. LC2 - Control diet plus 1.5 g/100 g OptiCell C5.

Table 3. Measurements of innate immunity (phagocytosis and oxidative burst of heterophils¹) and active immune function (proliferation of lymphocytes² and egg yolk IgY³) in Hy-Line Brown layer hens fed diets containing different types of lignocellulose fibre for 8 weeks from 20 weeks-of-age. Heterophil functions measured after being fed diets for 4 weeks, lymphocyte proliferation after 7 weeks and egg yolk IgY after 8 weeks. (mean, pooled SEM, n = 6).

Treatment	C	LC1	LC2	p Value	Pooled SEM
Heterophil functions
PI (latex beads/100 cells)	387.7^a	489.7^b	423.5^ab	0.006	14.6
%P (% cells containing 1 or more beads)	80.0^a	85.8^b	81.5^ab	0.027	1.0
Oxidative burst ΔRFU	1607.8	1656.4	1631.8	0.976	85.6
Lymphocyte proliferation^3
T lymphocytes (ConA stimulated)	82.8	95.4	95.1	0.257	3.5
B lymphocytes (LPS stimulated)	93.6	105.5	97.1	0.083	2.3
T and B lymphocytes (PMA stimulated)	160.8	182.2	178.9	0.767	12.3
Egg yolk IgY
mg/g yolk	8.96	9.96	8.93	0.543	0.42
mg/whole yolk	121.71	138.82	124.53	0.556	6.62

See Table 2 for information on treatment diets C, LC1 and LC2.

One hen/ pen was used for heterophil and lymphocyte measurements and for IgY a mean for each pen was obtained from 2 egg yolks.

^a,bMeans within the same row with different subscript letters are significantly different (p < 0.05).

1 - Oxidative burst measured 1 h after stimulation with phorbol 12-myristate 13-acetate as a change in relative fluorescent unit (ΔRFU) in hens fed experimental diets for 4 weeks.

2 – Mitogen stimulated lymphocyte proliferation in hens fed experimental diets for 7 weeks. Expressed as a percentage of the increase of mitogen stimulated lymphocytes relative to non-stimulated lymphocytes. Mitogens:- ConA - 2.5 μg/mL concanavalin A; LPS - 3.125 μg/mL lipopolysaccharide from Escherichia coli; PMA - 1.25 μg/mL phorbol 12-myristate 13-acetate.

3 - IgY measured in egg yolks from hens fed diets for 8 weeks.

Table 4. Egg production and egg quality measurements of Hy-Line Brown layer hens fed diets containing different lignocellulose fibre supplements for eight weeks from 20 weeks-of-age (means, pooled SEM, n = 6 pens).

Treatment	C	LC1	LC2	p Value (rows)	Pooled SEM (rows)
Weeks fed diets/Age (in weeks)		Production rate eggs/hen per d^c
0 – 2 / 20 − 22	0.73^A	0.75^A	0.74^A	0.761	0.015
2 – 4 / 22 − 24	0.88	0.91	0.90	0.513	0.009
4 – 6 / 24 − 26	0.90	0.94	0.92	0.196	0.008
6 – 8 / 26 − 28	0.91^aB	0.96^bB	0.94^abB	0.009	0.006
P Value	0.003	<0.001	0.001
SEM (columns)	0.034	0.010	0.030
		Whole egg weight including shell (g)^d
0 – 2 / 20 − 22	50.0^A	51.1^A	50.5^A	0.587	0.4
2 – 4 / 22 − 24	54.5	56.0	55.1	0.265	0.4
4 – 6 / 24 − 26	57.7^a	60.3^b	58.2^ab	0.031	0.4
6 – 8 / 26 − 28	59.6^aB	62.1^bB	60.1^abB	0.018	0.4
p Value	<0.001	<0.001	<0.001
SEM (columns)	0.501	1.046	0.286
		Egg quality after 4 weeks
Haugh unit^e	100.59^A	102.35^A	101.23	0.466	0.57
Shell thickness (mm)	0.404	0.408	0.407	0.956	0.01
		Egg quality after 8 weeks
Haugh unit^e	95.98^B	98.97^B	98.21	0.151	0.65
Shell thickness (mm)	0.402	0.427	0.412	0.073	0.005
p Value	0.027	0.025	0.147
SEM (columns)	1.484	1.070	1.760
Whole egg weight (g)^f	60.56	62.88	61.46	0.202	0.53
Egg shell weight (g)	5.93	6.12	5.92	0.748	0.12
Yolk weight (g)	13.44	14.16	13.92	0.430	0.22
Albumen weight (g)	41.19	42.60	41.62	0.435	0.44
RW egg shell	0.098	0.097	0.095	0.797	0.002
RW yolk	0.222	0.225	0.227	0.911	0.004
RW albumen	0.680	0.678	0.677	0.912	0.003

See Table 2 for information on treatment diets C, LC1 and LC2.

^{a, b}Means within the same row with different subscript letters are significantly different (p < 0.05).

^{A, B}Means within the same column for a parameter at 24 and 28 weeks-of-age with different uppercase superscripts differ significantly (p < 0.05).

RW: relative weight (g/100 g whole egg).

^cStrain average egg production rate at 20 – 22 weeks-of-age is 0.695 and at 26 – 28 weeks, 0.953 g (converted from values in hen-d percentage, Hy-Line Brown International (2014)).

^dStrain average egg weight at 20 – 22 weeks is 51.88 g and at 26 – 28 weeks-of-age it is 59.05 g (Hy-Line Brown International 2014).

^eHaugh units at 24 weeks of age average, 96.0 and at 28 weeks-of-age, 94.2 (Hy-Line Brown International, 2014).

^fAfter 4 and 8 weeks being fed experimental diets, the means of 2 eggs/pen were used to measure egg quality and weights of egg components.

Hy-Line Brown International. 2014. Hy-Line Brown management guide – Commercial layers. Iowa, USA: Hy-Line International; http://www.hyline.com.au/wp-content/uploads/2015/06/HL-Brown-Commercial-Manual-BRN-COM-ENG.pdf. Pp1–40. [accessed January 2014]

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Data availability statement

The data that support the findings of this study are available from the corresponding author (Dr. S. M. Hussein) upon reasonable request.