03 set peter ferket - cientifica potential of iof.pdf

The Potential of In Ovo Feeding and Early Nutrigenomic Programming

Peter R. Ferket N.C. State University

The Greek Philosopher's Genetic Theory “An individual’s traits are acquired from contact with the environment, and can be inherited by their offspring.”

You are Programmed to Succeed!

Poultry Breeding Based on Mendelian Genetics

Gene6cs assumes heritable traits are based DNA sequence

Mendel

85% of Improvement in Poultry Production Due to Genetics!

1956 2006 42 d, Live Wt, g 540 2805 6 wk Feed/Gain 2.35 1.70 Days to 1.8 kg +112 32

• >45 g weight/year • >1% increase/year (Havenstein et al., 2007)

Growth

Incuba\on Neonate Produc\ve Growth

Maintenance

In 1956, the incuba6on/neonate stage represented <25% of a broiler’s life

1956

Growth

Incuba\on Neonate Produc\ve Growth

Maintenance

But Now It is 50%!

Genetics changed the playing field…

…but expression of gene6c poten6al is what drives performance and profits

Nutrition has not kept pace with genetics

The time has come to

CLOSE THE GAP

Molecular Biology is Changing Our Understanding of Heritability

•  Study of gene expression by up-‐regula\on of mRNA

•  Study of Proteomics

•  Study of Metabolonics

Epigenetics = “On-Genes”

Epigene6cs assumes heritable traits are based on gene expression

Lamarck

It’s the link between gene6cs and nutri6onal adapta6on!

Swarming Locusts

EpigeneticEvidence

•  Swarming phenotype is environmentally influenced

•  Trait is passed genera\on to genera\on.

Exposure to an adverse s\muli during cri\cal

periods of development can permanently

reprogram normal physiological responses.

Early Life Programming

can turn on “Thrifty” Genes

Diego V. Bohórquez©

Why the difference between these two genetically identical bees?

Diego V. Bohórquez©

The first meal makes the difference!

Synchronize in-‐ovo feeding when the embryo begins to imbibe amnio6c fluid

(“The Avian Embryo”, Romanoff 1960)

Op6mum IOF Window

•  E17-‐E18 for Broilers •  E23-‐24 for Turkeys

Volume of amnio6c fluid (ml)

Days 1 3 5 7 9 11 13 15 17 19 21

8

6

4

2

IOF

•  Administration of highly digestible nutrients (CHO, Protein, Minerals, Vitamins, etc. ) into the amnion of late term embryos

•  Benefits: –  Advanced gut development –  Increased glycogen reserves –  Advanced muscle development –  Increase growth performance –  Better humeral and innate immunity –  Improved chick or poult quality –  Changes gene expression

Uni and Ferket, 2003. US Patent No. 6592878.

Jejunum Cross-Sections 20 Day-old Broiler Embryo

Control

In Ovo-‐Fed* * 1 ml. Saline containing 10% Maltose + 10% Sucrose + 5% Dextran

Control

IOF

Turkey Embryo Jejunum 25 Days of Incubation

Oliveira et al. (2008)

Turkey Embryo Jejunum At Hatch

Control

IOF


Control In-ovo Fed

Bohorquez et al. (2009)

Control In-‐ovo Fed

IOF advances villus development and func6on

of hatchling poults BohÓrqez et al. (2009)

In Ovo Feeding Increases Liver Glycogen

1 ml of Primary IOF solution given at 17 E

18E 19E 20E Hatch

Days before hatch

0

4

8

12

16

20

Glycogen

conten

t (mg/g o

f wet tissue

)

Control In-‐Ovo Fed

A A

A

A

A A

BB

18E 19E 20E HatchDays before hatch

0

4

8

12

16

20

Glycogen

conten

t (mg/g o

f wet tissue

)

Control In-‐Ovo

A A

A

B

A

A

B

B

Cobb Ross

In Ovo Feeding Increases Liver Glycogen Content in Poults

mg

Gly

coge

n

b

b

a

a

NC STATE UNIVERSITY


Day 3 Hatch

0 7E-‐8E 16E 21E

Embryonic myoblasts

Fetal myoblasts

Adult myoblasts-‐ Satellite cells

Day 8

in-ovo feeding

In ovo Feeding Timed to Boost 2nd and 3rd wave of satellite cell development

Skeletal Muscle Satellite Cells (Thymidine incorporation - CPM)

7 d of age 3 d of age Broiler Chicks 310 16,803 Non-‐injected 1,524 29,453 Commercial IOF 492% 175% ∆

Kornasio et al. (2011) PSJ

Control IOF

Hematoxylin & Eosin X40

Breast Muscle Fibers in 35d Broilers

Kornasio et al. (2011) PSJ

Effect of In Ovo Feeding on Body and Breast Muscle Weights (g) of Ross Broilers

Day 25 Day 10 Day of Hatch

In Ovo Control In Ovo Control In Ovo Control

997* 943 254* 243 47.0* 45.3 BW (g)

+ 5.7 + 4.2 +3.7 BW Diff. (%)

13.0* 12.0 12.3* 11.4 2.05* 1.93 Breast muscle % of BW

+8.3 +5.2 +6.2 Diff. (%) in Breast Muscle

•  1ml of Primary IOF solu6on given at 17 E. • Age of breeding flock:35 weeks

Uni et al. (2005)

Effect of IOF and Post-Hatch Feeding Time on Cobb (500)

Post-‐Hatch Fas6ng Time

In Ovo Treatment

2 d BW, g 35 d BW, g 0-‐35 d FCR

36 hr (SP) Control 38.7 1,995 1.63 36 hr (SP) IOF 42.0 2,209 1.69 6 hr (EF) Control 63.7 2,374 1.72 6 hr (EF) IOF 63.7 2,272 1.60 Source of Varia6on -‐-‐-‐-‐-‐-‐-‐-‐-‐-‐-‐-‐-‐-‐-‐-‐ (P-‐Value) -‐-‐-‐-‐-‐-‐-‐-‐-‐-‐-‐-‐-‐-‐-‐-‐-‐-‐-‐

Post-‐Hatch Fas\ng Time .0001 .0001 .974 In Ovo Treatment .0005 .185 .514 Feeding X IOF .0005 .001 .0485

(Kornasio et al., 2011)

Effect of IOF and Post-Hatch Feeding Time on Muscle

Development of Cobb (500)


Effect of IOF and Post-

Hatch Feeding Time on

Breast Muscle of Cobb (500)


•  Administration of highly digestible nutrients (CHO, Protein, Minerals, Vitamins, etc. ) into the amnion of late term embryos

•  Benefits: –  Advanced gut development –  Increased glycogen reserves –  Advanced muscle development –  Increase growth performance –  Better humeral and innate immunity –  Improved chick or poult quality –  Changes gene expression

Uni and Ferket, 2003. US Patent No. 6592878.

Effect of IOF on Bursa of Fabricius Development

•  IOF-treated chicks have larger bursa

•  Bursa has more and larger follicles

•  =more B cells ready to produce antibodies

∆

Comm.IOF

Non

Injected

+20.4%

0.177

0.147

Bursa to

Body weight

+18.2%

447

379

Follicle No. at

hatch

Uni et al. (2008)

Control In-ovo Fed

Bohorquez et al. (2009)

Mucin gene expression (MUC 2) in Chicks

Smirnov et al. (2006)

Control versus IOF:

Effect of Mineral Enrichment by IOF on Skeletal Development of Broilers

Control IOF Enriched (at E 18)

Amount per embryo Mineral / Vitamin concentration in the IOF solution

1.071mg Phosphorus (0.18%)

0.974mg Organic Iron (Bioplex) 0.16%

0.591mg Organic Zinc (Bioplex) 0.10%

0.36mg Organic Manganese (Bioplex) 0.06%

0.36mg Organic Calcium (Bioplex) 0.06%

0.018mg Organic Copper (Bioplex) 0.003%

136.8IU Vitamin D3 0.001%

(Yair and Uni, 2011)

Levels and uptake of minerals from the yolk in control and in ovo mineral

enrichment


Effect of IOF of Organic Minerals on Tibia Mechanical Properties of Broilers

The enrichment resulted in improvement in the mechanical properties of tibiae.


Blue=down regulated

Red=up regulated

Method = Ward

HBZ

HOXA3

CTGF

PDK1

SOD1

MYBPC3

TOP2B

SM22

AZIN1

TLR4

HSPE1

HB alphaA

EDNRA

ANF / ANP

HBE

IGFBP1

EFNB2

Tenascin precursor

Signal transducer and activator of transcription 3

Beta-actinCytoskeletal beta actin

GAPDH

CHICK DNA

TURKEY DNA

fructose biphosphatase (F1,6 BPase)

transketolase

creatine kinase

aconitate hydratase

malate dehydrogenase

maltase-glucoamylase

glutamate dehydrogenase

glyceraldehyde 3-phosphate dehydrogenase

growth hormone receptorphosphopyruvate hydratase

glycogen synthase 1 (muscle)

isocitrate dehydrogenase (NADP+)

carnitine O-palmitoyltransferasegrowth hormone receptor (GHR)

succinate dehydrogenase

EF2

CHICK DNATURKEY DNAaminopeptidase Ey

Std

Lsm

ean

Day

25

Std

Lsm

ean

Day

26

Std

Lsm

ean

Day

28

Std

Lsm

ean

Trt n

o

Std

Lsm

ean

Trt y

e

Std

Lsm

ean

Trt*

Day

no

25

Std

Lsm

ean

Trt*

Day

no

26

Std

Lsm

ean

Trt*

Day

no

28

Std

Lsm

ean

Trt*

Day

ye

25

Std

Lsm

ean

Trt*

Day

ye

26

Std

Lsm

ean

Trt*

Day

ye

28

Dendrogram

Hierarchical Clustering of Standardized LSMeans

Genes with similar pattern were grouped in the same cluster/color

43/120 genes expressed differently over \me by IOF


Hori zontal reference l ine drawn at -log10(p ) = 4.01Frame Si ze for Gra phs BelowMarker Size for Graph s Below

0

10

-log1

0(p-

value

) for

Diff

of T

rt =

(CO

N)-(I

OF)

HOXA3

PA

MYL4

PLB

HIF1AACTA2

IFN? HMOX1

MYL9

EGFr

IGFBP1TLR2 -2IL13

Prepro endothelin -1

LEREPO4 protein - simila r

IL-8

FasL

creatin e ki nase

maltase-g lucoamylasehexokinase 1

fructose b iph osphate al dolase B

glycogen synthase kin ase-3 beta

aminopep ti dase

tyrosinase

F_actin

CDK1

14_3_3

-0.5 -0.4 -0.3 -0.2 -0.1 0 0.1 0.2 0.3 0.4 0.5Diff of Trt = (CON)-(IOF)

Diff of Trt = (CON)- (IOF)

Diff of treatment: Control -‐ IOF

Fold difference: 4.0 -‐ Log

10 (P-‐value

) of D

iff

Bohorqez et al. (2009)

•  EGFr •  ILGF-BP1 •  Thyroid receptors α1 & β1

Epithelial growth

•  IL-8 •  TLR2 •  Fas-Ligand

Innate Immunity

•  Maltase-glucoamylase •  Glycogen synthase kinase •  Aminopeptidase 1

CHO and Protein Metabolism

•  F-actin capping protein •  Myosin light-chain 4 & 8

Smooth Muscle Development

Bohorqez et al. (2009)

Gene Expression Patterns: The Powerful Tool of Transcriptomics Decoding nutri6onal control: Nutrigenomics

Bridging the gap between

genetics and nutrition

Early nutri6on can imprint desired metabolic traits.

Imprint for the desired trait when they are young

Nutritional Imprinting

What Production Traits Can Be Imprinted by Adaptive Conditioning of Gene Expression?

•  Tolerance to immunological stress •  Tolerance to environmental stress •  Tolerance to oxida\ve stress •  Energy u\liza\on and requirement •  Mineral u\liza\on and requirement

It is not just what we feed, but also when we feed it!

The first 90 hours of the chicks life

determines a bird’s ability to use specific nutrients

Nutritional Conditioning

Heat map shows differential gene expression patterns in intestinal tissues from broilers at 21 days

Control Condi6oned Uncondi6oned

The conditioned birds have totally different physiological baseline

Effects of the Programmed Nutrition (PN) Strategy on Ca

and P Digestion in Broilers Diet Ca Diges6on (%) P Diges6on (%) Corn-‐soy control diet 23.9b 23.1c

PN supplements without condi\oning 28.6b 28.8b

Complete PN strategy 44.6a 39.7a

Alltech-‐UK Nutri\on Research Alliance (2011)

Enhanced Mineral Retention a Using PN Strategy Mineral Content in Breast Muscle

Mineral content Control Alltech PN diet Calcium (ppm) 41 44 Selenium (ppb) 120 332 Zinc (ppm) 3.04 3.03 Manganese (ppb) 129 157 Copper (ppb) 513 457 Iron (ppm) 3.31 3.58

Note: PN diets contain up to 50% less Zn, Mn, and Cu

(Data from Alltech-‐UK Nutri\on Research Alliance, 2011)

Improved Meat

Quality Item Standard

diet Programmed Nutri6on

Total an\oxidant capacity (µMol)

1.76 1.85

Color (red index) 9.12 9.77 Drip loss (%) 2.19 1.32 pH 6.03 6.15

Oxidative Stability: Freshness

Quant et al. (2011) Alltech-‐University of Kentucky Nutri8on Research Alliance, Lexington, KY

Rancidity

Programmed Nutri6on

Control

Breast Filet Cooking Characteristics

Cooking characteris6cs Diet Cooking loss, % Control 17.05 Programmed Nutri\on 15.27 SEM 0.92

Potential Benefits of Nutrigenomic Programming • Nutrient U\liza\on •  Performance efficiency •  Disease Resistance • Meat yield and quality •  Reproduc\ve efficiency •  Enteric ecosystem •  Behavior

This is only the beginning!

It is the next Step Forward in Nutrition.

03 set peter ferket - cientifica potential of iof.pdf

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