EFFECT OF A MICROBIAL 6-PHYTASE SUPPLEMENTATION ON PLASMATIC MYO-INOSITOL AND ON THE PECTORALIS MUSCLE TRANSCRIPTOME IN BROILERS

J. Schmeisser1, R. Aureli1, A. Friedel2, A-A. Séon1, P. Guggenbuhl1, A. Cowieson2 and F. Fru.

1DSM Nutritional Products France, Centre de Recherche en Nutrition Animale, BP 170, 68305 Saint-Louis Cedex, France2DSM Nutritional Products Ltd, Kaiseraugst, Switzerland

Phytase efficiency to increase phosphorus utilization inpoultry has been proven for decades. In addition,phytase was demonstrated to improve growthperformance, meat breast weight, amino acidsdigestibility and plasma myo-inositol concentration.The objective of this work was to investigate potentialinteractions between phytase supplementation, growthperformance and host gene expression to identifypotential associated biomarkers.

Animals:

Diets:

Parameters:

- 16 broilers per treatment fed a corn/soybean mealbased diet ad libitum 36 days

- Negative control (NC) - Pi deficient (Total P=0.44%)- Positive control (PC) – Pi sufficient (Total P=0.75%)- NC + RONOZYME® HiPhos at 1000 U/kg- Transcriptomic analysis of breast muscle (Pectoralis

major) with Affymetrix CHIGENE-1.0-ST arrays(4 pools of 4 samples per treatment) and validationof selected genes by Q-PCR (n=16).

• Specific biomarkers potentially related to phytase supplementation havebeen identified in muscle.

• The results highlighted the importance of IP3 and myo-inositol (end productsof phytic acid degradation by phytase) for improving muscle growth.

• Confirm these results in other farm animal species and study the host geneexpression pattern in relation to other feed enzymes.

Most impacted pathways in animals fed RONOZYME® HiPhos:• Development - Calcium/calmodulin-dependant kinase and IGF

(via myo-inositol stimulation) controlling skeletal myogenesis

Number of modulated genes compared to negative control(with a minimum fold-change of 1.3 or -1.3 and p<0.05):

Fold-change compared to negative control

Microarray (n=4x4) Q-PCR (n=16)Gene symbol

HiPhos1000 PC

HiPhos1000 PC

PPP3CBMEF2AMEF2CPLCB1PI3KR1

1,67*1,44*1,43*1,432,02**

1,181,081,171,021,32

2,67*1,68*2,14*2,66*3,04*

1,881,421,731,481,51

Calcium/calmodulin-dependant kinase and IGF controlling skeletal myogenesisand IP3 signaling

* p< 0,05 ** p< 0,01

IGF-1receptor

IGF-1

PIP2

PIP3

PIP2IP3

CalciumCalmodulin (CALM1)

Myocyte enhancerfactor 2 (MEF2)

K(lysine)acetyltransferase 2B

(KAT2B)

Myogenic regulatory factor

(MYOD)

SKELETAL MUSCLE HYPERTROPHY SKELETAL MUSCLE DIFFERENCIATION

Myo-inositol

Myo-INO Phosphoinositide-3-kinase(PIK3R1 & PIK3CA)

Protein phosphatase 3 (PPP3CB)

Calcineurin A

IP3 receptor3-phosphoinositide dependent protein kinase-1 (PDPK1)

Pkb/AktPathway

Ribosomal protein S6 kinase b-1 (RPS6KB1)

PROTEIN SYNTHESIS

IGF binding protein (IGFBP)

Phospholipase C bêta (PLCB)

Up-regulated genes with phytase

PC vs NC HiPhos1000 vs NC205 53095

Confirmation of expression of selected genes by Q-PCR:

0

500

1000

1500

2000

2500

3000

PC NC HiPhos 1000

Wei

ght

gain

(g)

0,050,0

100,0150,0200,0250,0300,0350,0

Brea

st w

eigh

t (g

)

PC NC HiPhos 1000 0,010,020,030,040,050,060,070,0

Myo

-ino

sito

l (m

g/L)

PC NC HiPhos-1000

Myo-inositol effects on insulin sensitivity

ab b

a aaa

bb

a,b: Means within the chart with common superscripts do not differ significantly (p<0.05)