recommended vitamin levels for high performance broilers and layers - gilberto litta

Recommended Vitamin Levels for High

Performance Broilers and Layers

IV Simposio Internacional Sobre Exigencias Nutricionais de Aves e Suinos

Universidade Federal de Vicosa, Vicosa, Brazil March 29th – 30th , 2017

Gilberto Litta

Animal Science & Advocacy Manager

DSM Nutritional Products, Kaiseraugst (Switzerland)

The Century of the Vitamins

1900 20171950

1929-1967: 12 Nobel Prices

awarded to 20 Scientists for

Synthesis, Characterization

und Elucidation of Function of

the Vitamins

1912: Term

“Vitamins”

coined by

Casimir Funk

1946: First Large-

scale, industrial

Synthesis of Vitamin A

by Roche

1934-87: Development of industrial

Production Processes for Vitamins by Roche,

starting with Vitamin C

2016: 13th Edition of the

Vitamin Recommendation Folder

DSM Vitamin Supplementation

Guidelines for Domestic

Animals published

1906-41: 13 Vitamins

identified and characterized

▪ Fat soluble vitamins (4):

▪ vitamin A (retinol) vitamin D (calciferols)

▪ vitamin E (tocopherols) vitamin K (phylloquinone)

▪ Water soluble vitamins (9):

▪ vitamin B1 (thiamin) vitamin B2 (riboflavin)

▪ vitamin B6 (pyridoxine) vitamin B12 (cobalamin)

▪ niacin (B3;PP) pantothenic acid (B5)

▪ folic acid (B9; M) biotin (B7; H)

▪ vitamin C (ascorbic acid)

VitaminsClassification and Nomenclature

▪ Choline is sometimes included in the list;

▪ Inositol is essential for aquatic species only

▪ Vitamins are essential micronutrients, required for optimum

health and normal physiological functions such as growth,

development, maintenance or reproduction of the animal.

▪ Vitamins exercise catalytic functions; they facilitate both

synthesis and degradation of the nutrients, thereby controlling

the metabolism.

▪ Most vitamins cannot be synthesized by the animals and

therefore they must be obtained from the feed.

Vitamins: Definitions and Functions

Functions of VitaminsVitamin Basic function(s) Deficiency disorders/diseases

Vitamin A photosensitive retinal pigment, regulation of epithelial cell

differentiation, regulation of gene transcription

blindness, xerophthalmia,

keratomalacia, impaired growth

Vitamin D promotion of intestinal Ca absorption, mobilization of Ca from

bone, stimulation of renal Ca resorption, regulation of PTH

secretion, possible function in muscle

Rickets, Osteomalacia

Vitamin E antioxidant protector for membranes nerve, muscle degeneration

Vitamin K co-substrate for γ-carboxylation of glutamyl residues of several

clotting factors and their Ca-binding proteins

impaired blood clotting

Vitamin B1 coenzyme for oxidative decarboxylation of 2-keto acids,

coenzyme for pyruvate decarboxylase and transketolase

Beriberi, polyneuritis, Wernicke-

Korsakoff syndrome

Vitamin B2 coenzyme for numerous flavoproteins that catalyze redox

reactions in fatty acid synthesis/degradation, TCA cycle

dermatitis

Vitamin B6 coenzyme for metabolism of amino acids symptoms vary by species

Vitamin B12 coenzyme for conversion of methylmalonyl-CoA to succinyl-

CoA, methyl group transfer from 5-CH3-FH4 to homocysteine in

methionine synthesis

megaloblastic anemia, impaired

growth

Pantothenic

Acid

co-substrate for activation/transfer of acyl groups to form

esters, amides, citrate, triglycerides, etc.

symptoms vary by species

Niacin co-substrate for hydrogen transfer catalyzed by many

dehydrogenases, e.g. TCA cycle respiratory chain

Pellagra

Folic Acid coenzyme for transfer of single-carbon units megaloblastic anemia

Biotin coenzyme for carboxylations dermatitis, cracked hooves

Choline component of acetylcholine and the membrane structural

component phosphatidylcholine

poor growth, Perosis (deformity of

leg bones in young birds), fatty

liver

Vitamin C co-substrate for hydroxylations in collagen synthesis, steroid

metabolism

Scurvy

• Vitamins can almost never be regarded as nutrients in isolation

given that they display a wide range of interactions

▪ Fat soluble vitamins must be fed in correct ratios as they all

compete for intestinal absorption

▪ Water soluble vitamins are regulators of the intermediary

metabolism of protein, fats and carbohydrates (energy): a lack

of one of them increases the need of the others

• It’s very difficult to estimate precisely the optimum

requirements for maximizing performance for each vitamin

separately and mathematical relationships can’t be established

(Whitehead, 1987)

VitaminsFunctions and requirements

Metabolic functions and interactions of B-

group vitamins

Amino acid

pool

B6 FA B12 Krebs (citric

acid) cycle

B2 PP B1

B1 = Thiamin

B2 = Riboflavin

PP = Nicotinamide

B6 = Pyridoxine

B12 = Cobalamine

H = Biotin

FA = Folic acid

PA = Pantothenic acid

Amines

Purines

Pyruvic

acid

Glucose

B6

FA

B2B6

Proteins Carbohydrates

Uric acid Urea

B6

B6

Glycogen

Glycerol Fatty acids

Acetyl CoA

B1B2

PP

Fats Diet

Cell

Cell

Excretion

B2

PP

PAB2 PP H

PPB1

B Vitamins and Methionine cycle

MethionineHomocysteine

Vitamin B12

B9, CH3-THF

SAM - SAH

Choline Betaine

CH3 acceptor (nucleic acid, protein & amine synthesis, phosphatidylcholine, creatine

Cysteine

Vitamin B6

☺

Vitamin

Demand

Housing

Conditions

Genetics/

Breed

Feed

Composition

Life Stage

Performance

Stress

Infectious

Pressure

Temperature

/Humidity

Factors influencing Vitamin Requirements

Comparison of Ross broiler live weight for age over time (as hatched)

Source: Ross Performance and Management Guide, 1980, 2007 and 2014

Genetic driven improved performance requires each year

an adjustment of vitamin supplementation in the range of 1%

The improved efficiency reduces vitamin intake per unit of productivity

and dictates the need for more dietary vitamin

1985 2005 ∆/year

Layer (1 kg) 2.7 IU/egg 2.1 IU/egg -1.1%

Broiler (2 kg) 40 IU/kg gain 34 IU/kg gain -0.8%

Turkey (14 kg) 55 IU/kg gain 48 IU/kg gain -0.6%

All diets containing 20 IU Vitamin E/kg feed

Impact of Genetic Improvement

on Vitamin Supplementation

Source: S. Leeson, World’s Poultry Sci. Journal, 2007 63 255-266

minimum

optimum

adequate

▪ Avoid clinical (and sub-clinical?) deficiency symptoms

▪ Generate maximum performance and feed utilization

▪ Satisfy needs for efficient nutrient metabolism

▪ Maintain adequate vitamin plasma and tissue levels

▪ Enable successful reproduction

▪ Support optimum health and welfare

▪ Develop superior product quality

Criteria of Vitamin Requirements

Increased Blood-clotting TimeDepression of Appetite

Resorption DisordersInflamed MouthSusceptibility to Infection

Hepatic NecrosisPododermatitisOsteomalacia

Cervical ParalysisFatty Liver and Kidney Syndrome

Muscular MyopathyNecrosis of Heart FibersPerosis

DiarrhoeaAscitesPoor Absorption of Nutrients

Deformed /Brittle BonesLow Immune Response

Fertility ProblemsParalysis / Lameness

What is the Reason for Production

Problems such as …

Vitamin Deficiencies

Dry / Scaly Skin

Encephalomalacia –Vitamin E (and Selenium)

Rickets - Vitamin D3 Perosis - Biotin

Vitamin B1 Deficiency Vitamin B2 Deficiency Fatty Liver Hemorrhagic Syndrome

(FLHS) Vitamin B12 and E Deficiencies

Vitamin Deficiencies in Poultry

▪ Inadequate vitamin intake with the feed:

▪ low natural vitamin levels in feedstuffs, low availability

▪ presence of vitamin antagonists (avidin [egg white] biotin)

▪ insufficient vitamin supplementation levels / mixing errors.

▪ Poor digestion and absorption of vitamins

▪ gut health issues impair fat soluble vitamins absorption

▪ Increased vitamin requirements:

▪ depending on diet composition (e.g. vitamin E for PUFA’s)

▪ for immune response and during phases of stress or diseases.

Causes of Vitamin Deficiency

Vitamins and…

1. Bone development and health

2. Immune system modulation

3. Gut health

4. Stress

5. Performance

6. Meat and egg quality

Bone development and health

▪ Vitamina D3

▪ 25OHD3

▪ Vitamina K

▪ Vitamin C

0,0

0,5

1,0

1,5

2,0

2,5

3,0

3,5

7 14 21 28 35 42

Gut

Lesi

on S

core

(1=m

ild;

2=m

od;

3=se

vere

Average Age (days)

Histopathic Evidence Day 14-21

Gross Evidence Day 21-28

Legs Day 28-finish

Malabsorption-induced lameness

Malabsorption reduces efficiency of vitamin D metabolism (intestinal

absorption) with direct negative impact on bone development/health….

….and bone health is also impaired by the immune response…

Diseases and immunological aspects antagonize production functions

Anorexia and Fever (IL-1,

TNF-a)

Increased skeletal muscle protein

degradation (IL-1)

Infective disease

Cytokine production

(TNFα, IL-1b, and IL-6)

Acute phase protein response

Decreased Body Weight Gain and Muscle Growth

Lymphocyte/Macrophage

stimulation

Decreased bone deposition and Increased bone resorption

Increased lipolysis in adipocytes (IL-1,

TNF-a)Increased release of

corticosterone (IL-1, IL-6)

Cytokines promote bone resorption

TNFα, IL-1b and IL-6 are cytokines promoting osteoclast* synthesis and activity

TNFα, IL-1b and IL-6

* Osteoclast are the cells in the bone tissues responsible of bone resorption

(i.e.«destroying» bone tissue) for mobilizing Ca++ from bones to blood

Trait

(Ca/P: 8.0/3.5 g/kg)

Vitamin D3

200 IU/kg

Vitamin D3

800 IU/kg

Vitamin D3

5,000 IU/kg

Vitamin D3

10,000 IU/kg

Experiment 1

Liveweight 315a 311a 316a 336b

Tibia breaking

strength

61.0a 78.1b 90.9c 93.5c

TD incidence 88a 51b 6c 8c

Experiment 2

Liveweight 295a 297a 303a 351b

Tibia breaking

strength

36.4a 44.5a 61.4b 76.1c

TD incidence 78a 84a 22b 0b

Source: Whitehead et al., 2004

Effect of Vitamin D3 on Bone Structure and Functionality

Skin

Liver

Kidney

hydroxylation

hydroxylation

Cholecalciferol (Vitamin D3)

25-hydroxycholecalciferol

(25-OH-D3, Circulating form)

1,25-dihydroxycholecalciferol

(Active form)

Skin

Liver

Kidney

7-dehydrocholesterol Dietary Vitamin D3

UVB

irradiationSkin

Liver

Kidney

hydroxylation

hydroxylation

Cholecalciferol (Vitamin D3)

25-hydroxycholecalciferol

(25-OH-D3, Circulating form)

1,25-dihydroxycholecalciferol

(Active form)

Skin

Liver

Kidney

7-dehydrocholesterol 25OHD3 (Hy•D®)

Vitamin D3 and 25OHD3 metabolism

UVB irradiation

(280-315 nm)*Absorption occurs

virtually independent of

fat digestion

Source: Rebel A. And G. Weber, 2009, XVI World vet. Poultry Congress, Marrakesh

14,71

14,2712,96

29,98

37,67

36,84

9,88

0 0

19,87

14,0416,17

0

5

10

15

20

25

30

35

40

3 d p.i. 6 d p.i. 8 d p.i.

25O

HD

3 p

lasm

a (

ng/m

l)

Days post-infection

Uninfected Control Uninfected Hy•D®

Infected Control Infected Hy•D®

* Day-old broiler chicks were inoculated either with saline (Uninfected) or with MAS (Infected) on the day of hatch.

Control: vitamin D3 2.760 IU/kg feed; HyD: 69 mg 25-OH-D3/kg feed

Effect of Malabsorption Syndrome (MAS) on 25OHD3

serum level

25OHD3 vs Vitamin D3 effect on bone strengthSingle trials results 2003 - 2012

25OHD3 (Hy•D®) improved bone strength 10,8% against vitamin D3

Source: summary of experimental and field trials 2003 - 2012

4,5%

18,5% 18,1%

9,5%

5,7%

1,5%

4,8%3,5%

29,9%

17,2%

5,6%

10,8%

0,0%

5,0%

10,0%

15,0%

20,0%

25,0%

30,0%

35,0%

Bone Strength % improvement HyD vs Vitamin D3

Page 23

Effects of vitamin D3 or 25OHD3 on the incidence of TD, Head Femur Necrosis and Valgus Various defects

0

10

20

30

40

50

60

70

80

90

Control Hy·D® Control Hy·D® Control Hy·D® Control Hy·D®

21 - 28 29 - 35 36 - 42 43 - 49

Tibial Dyschondroplasia, %

0

20

40

60

80

100

120


21 - 28 29 - 35 36 - 42 43 - 49

Head Femur Necrosis, %

0

5

10

15

20

25

30

35

40


21 - 28 29 - 35 36 - 42 43 - 49

Valgus Various defects, %

25OHD3 in the feed reduced

bone disorders vs vitamin D3

weeks

Source: Naas et al., 2012

25OHD3 vs Vitamin D3 and Tibial Dyschondroplasia

0

2

4

6

8

10

12

TD Severity TD Incidence

% a

ffecte

d B

irds

D3: 2’760 IU/kg Hy•D: 68.9 µg/kg

Hy•D: 344 µg/kg

AUBURN UNIVERISTYROSLIN INSTITUTE

Control D3

(75 µg/kg)

Hy•D

(75 µg/kg)

Normal Tibial

Growth Plates:35.0% 88.0%

Abnormal Tibial

Growth Plates:

Severity 0 25.0% 2.0%

Severity 1 12.5% 2.0%

Severity 2 16.7% 6.0%

Severity 3 10.4% 2.0%

100.0% 100.0%

75 mg/kg = 3,000 IU/kg

0

5

10

15

20

25

30

35

40

45

50

28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55

Cum

ula

tive lam

eness

(n.

case

s )

Days

Control HyD

25OHD3 (Hy•D®) reduced late lameness incidence against vitamin D3

Control diet with vitamin D3 5.500 IU/kg; HyD 69 mg/kg in the water on top of control diet

Late Lameness Study

Source: Wideman et al., Poultry Sci., 2015

Pullet Development

Nutrition of pullets is critical for building a strong, healthy and high yielding laying hen,

especially for bones

Source: adapted from Bregendhal K., Nutrition of laying hens, Hy-Line

Immune and digestive

system and bones

Bones, muscles &

feathers (frame)

Oviduct and

medullary bone

85% of the skeleton

develops within 6

weeks and 95%

within 10 weeks

CONFIDENTIAL

Vitamin D3 vs 25OHD3 (Hy•D®) on mineralapposition rate in pullets

Vitamin D3

(2,760 IU/kg)

Vitamin D3

(5,520 IU/kg)

Vitamin D3 (2,760 IU/kg) + Hy•D® (69 µg/kg )

Mineral apposition rate: Calcein labeling technique

Sourec: Kim et al. 2016, University of Georgia (preliminary data, unpublished)

25OHD3 vs Vitamin D3 effect on bone health in layers

• Hy•D® increased bone strength (greater cortical bone density)

• Hy•D® groups used more efficiently the medullary bone instead of destroying the cortical tissue

(structural bone)

Source: Korver & Saunder-Blades, 2005

The greater the cortical bone the lower the risk of bone fractures and cage layer fatigue

(with a decrease in egg production and lower shell quality)

0

50

100

150

200

250

0 160 320 640 1280 2560

Without Vitamin C

100 mg Vitamin C/kg feed

Effect of Vitamin C on Plasma Concentrations of 1,25OH2D3

Vitamin D3 (IU/kg feed)

pg/m

l pla

sma

0

5

10

15

20

25

0 160 320 640 1280 2560

Without Vitamin C


Effect of Vitamin C on the Intestinal Ca-binding Capacity


% b

indin

g

0

50

100

150

200

250

0 160 320 640 1280

Without Vitamin C


New

ton

Breaking Load of Tibia as Affected by the Supplementation of Vitamin C


Biotin and Pododermatits

Biotin Group (2000 mcg/kg) Control Group (200 mcg/kg)

Source: Buda et al., 2000

Surface of Reticulate Scales at Digital Foot Pad

of Turkeys

Control Group (200 mcg/kg)Biotin Group (2000 mcg/kg)

Surface of the Stratum Corneum at the

Reticulate Scale of Turkeys

Source: Buda et al., 2000

Immune system modulation

▪ Vitamina A

▪ Vitamina E

▪ Vitamina C

▪ Vitamina D3

▪ 25OHD3

The immune system

Vitamin A and cell-mediated immune responseFlow cytometric analysis of intraepithelial lymphocytes

Source: Dalloul et al., 2002 Poultry Sci.

▪ Pale grey bars: 8.000 IU/kg vitamin A;

Dark grey bars: vitamin A deficient

diet

▪ Lymphocytes expressing the surface

markers CD3,CD4,CD8,αβTCR, and

γδTCR, as well as surface IgA are

reported as the percentage of total

lymphocytes

Vitamin A deficiency, altering the

lymphocytes subpopulations,

reduced local cell-mediated

immunity and lowered the ability

of broilers to resist

Eimeria infection.

• Female broiler chicks (day-old)

• 4 dietary treatments:

• Basal diet (20 ppm Vit E)

• Basal diet + 100 ppm Vit E



• Vaccination at 28 days of age with

inactivated and emulsified

Newcastle disease virus

• Blood sample on days 7, 14, 21,

28, 38, 48 and 58

1,0

1,5

2,0

2,5

3,0

3,5

4,0

4,5

5,0

7 14 21 28 38 48 58H

I Anti

body T

iters

(lo

g2)

Days after Vaccination

Basal Diet BD + 100 ppm Vit E

BD + 200 ppm Vit E BD + 300 ppm Vit E

Adapted from Franchini et al., 1986 Clinica Vet.

Vitamin E modulates humoral immune

response in broilers

Vitamin E supplementation induced a higher production of antibodies

Group 1 2 3 4 5 6 7 8 9 10

Vitamin E,

mg/kg0 0 0 0 15 15 50 50 200 200

Vaccine BI No No Yes Yes Yes Yes Yes Yes Yes Yes

+ Virus Yes No No Yes No Yes No Yes No Yes

Group 1 2 3 4 5 6 7 8 9 10

Vitamin E,

mg/kg0 0 0 0 15 15 50 50 200 200

Vaccine BI No No Yes Yes Yes Yes Yes Yes Yes Yes

+ Virus Yes No No Yes No Yes No Yes No Yes

Bacteria count Nitric oxide (µM) produced by macrophages

Vitamina E and macrophages bactericide activity against Salmonella enteritidis

Fonte: Almeida et al., 2014

Vitamina C and immune response in broilers vaccinated against Gumboro

Parameters Control

no Vitamina C

+ 1000 ppm

Vitamin C

IgM+ in bursa (N)

Pre-vaccination (7 days) 65.32 76.24*

Post-vaccination (21 days) 82.82 92.00*

Post-adminstration of virus (31 days) 56.68 66.68*

Antibodies IgG production

(x106 cells)

Post-vaccination (21 days) 19.6 47.6*

Post-administration of virus (31 days) 29.6 37.6*

Fonte: Wu et al., 2000

* P<0.05

CONFIDENTIAL

Hy•D® benefits (Non-classical)

▪ Independent of calcium

metabolism

▪ 1α-hydroxylase enzyme

activity in immune cells

▪ Locally converts circulating 25-

(OH)D3 to 1,25-(OH)2D3

▪ Activity depends on adequate

circulating 25-(OH)

▪ VDR is present in immune

system cells

▪ 1,25-(OH)2D3 binds to VDR and

initiates responses

Sources: Norman, 2008; Shanmugasundaram & Selvaraj, 2012;

Morris et al., 2014, Shojadoost et al., 2015

Vitamin D3 and 25OHD3 and immune function

1a-Hydroxylase mRNA amounts in

different organs in chicks at hatch

0

10

20

30

40

50

60

70

80

Rela

tive m

RN

A a

mounts

(A

U)

CONFIDENTIAL

Saunders-Blades and Korver, Pre-Symposium XXIII World’s Poultry Congress, Brisbane, 2008

Early (~32 weeks)

25OHD3 fed to breeders significantly increased

chick’s mature innate immune function*

Mid (~47 weeks) Late (~62 weeks)

* Measured through white blood cell phagocytosis

Effect of Vitamin D3 and 25OHD3 on immune

modulation of breeders and chicks

3. Five poultry studies between 2012 & 20154 published and 1 in press

CONFIDENTIAL

Morris et al., 2014

Immune modulation▪ Challenge model

▪ Injection of LPS (lipopolysaccharides)

to cause inflammatory responses in broiler chickens

▪ Objective

▪ To evaluate Hy•D® effects in suppressing these responses

▪ Results

▪ Hy•D® improved weight gain compared to D3 post LPS challenge

▪ Exerted anti-inflammatory effects

▪ Beneficial during immune challenges

▪ Beyond starter supplementation maintained the effects

▪ Starter only supplementation did not

Vitamin D3 and 25OHD3 effects on immune modulation

in chickens challenged with LPS injection

-2

-1

0

1

2

3

4

5

6

Fold

change f

rom

the

chole

calc

ifero

l su

pple

mente

d

gro

up

Cholecalciferol Cholecalciferol + LPS

25-OH Cholecalciferol 25-OH Cholecalciferol + LPS

**

***

*

P values : Diet, P = 0.01; LPS, P = 0.87; Diet*LPS, P < 0.01.

25OHD3 supplementation decreases IL-1β mRNA

amounts in the liver post-LPS injection

Source: Morris A. et al. Poultry science, 2014

Treatment

**

Lipopolysaccharide is a potent immunogen in stimulating an inflammatory response in both in vivo

and in vitro experiments.

Lipopolysaccharide stimulation leads to release of inflammatory cytokines IL-1β and tumor necrosis

factor alpha (TNFa)

IL-1b: pro- inflammatory marker

12 h 48 h

0

1000

2000

3000

4000

5000

6000

7000

8000

9000

10000

Fold

change f

rom

the c

ontr

ol gro

up

*

* ** **

**

*

*

*

P values: 12 h: 25-hydroxycholecalciferol, P = 0.17; LPS, P < 0.01; 25-hydroxycholecalciferol *LPS, P = 0.25; 48 h: 25-

hydroxycholecalciferol, P < 0.01; LPS, P < 0.01; 25-hydroxycholecalciferol *LPS, P < 0.01.

25OHD3 supplementation increased IL-10 mRNA in

LPS stimulated macrophages

Source: Morris and Selvaraj, 2015

IL-10: anti- inflammatory marker

*

0

1

2

3

4

5

6

7

8

21 d 35 d

24 h

body w

eig

ht

gain

expre

ssed a

s % o

f pre

-LPS b

ody w

eig

ht

Cholecalciferol 25-OH Cholecalciferol 25-OH Cholecalciferol-14 d

Effects of 25OHD3 supplementation on broiler body

weight gain at 24 h post-LPS challenge

Source: Morris A . et al. Poultry science, 2014

P = 0.03

P = 0.01

**

**

*

***

*

+2,5% more BW gain

+3,8% more BW gain

P values: P < 0.01(linear), P = 0.20 (Quadratic)

50,36

130,1107,05

215,03

312,46

Nit

rite

(µM

)

25OHD3 treatment increased nitric oxide (NO) production

in LPS stimulated macrophage cells

Macrophages secrete a variety of inflammatory mediators and cytokines like TNFa-IL-1, ROS (Reactive Oxygen Species)

and NO (Nitric Oxide)

ROS and NO act as antimicrobicidal compounds responsible for killing pathogenic bacteria

Nitric oxide: immune response aspect

Source: Morris and Selvaraj, 2015

*

* *

**

**

CONFIDENTIAL

Morris et al., 2014

Immune modulation▪ 25OHD3 supplementation

▪ Decreased expression of IL-1β (marker for

inflammation)

▪ Increased expression of IL-10 (anti-inflammatory

properties

▪ Increased production of nitric oxide (antimicrobial)

▪ Modulating immune response 25OHD3 limited

weight loss during immune challenge

Conclusions

Stress

▪ Vitamin A

▪ Vitamin E

▪ Vitamin C

1,72

1,74

1,76

1,78

1,80

1,82

1,84

1,86

1,88

1.560

1.580

1.600

1.620

1.640

1.660

1.680

1.700

1.720

1.740

0 100 200 300 400

Feed C

onvers

ion

Weig

ht

Gain

Vitamin C, ppm

Weight Gain, g Feed Conversion

Broilers performance (1 to 42 days ) under heat stress and with grading levels of vitamin C

Effect of grading dosages of Vitamin E on egg yield of hens under heat stress

25 mg vit E/kg

125 mg vit E/kg

250 mg vit E/kg

a,b P<0,05

Fonte: Panda A. K. et al., 2008

84

84,2

84,4

84,6

84,8

85

85,2

85,4

85,6

85,8

86

86,2

Egg production (%)

b

aa

Broilers performance (1 to 42 days ) under heat stress and with grading levels of vitamin E

1,64

1,66

1,68

1,70

1,72

1,74

1,76

1,78

1,80

1.350

1.400

1.450

1.500

1.550

1.600

1.650

1.700

1.750

1.800

0 75 150 225 300

Feed C

onvers

ion

Weig

ht

Gain

Vitamin E, ppm

Weight Gain, g Feed Conversion

a,b,c P≤0.001; (vitamin E x temperature 0.001)

3,56

4,895,82 5,58

4,02

Tota

l anti

body (

log2)

Vitamin E (mg/kg) Temperature

92,4

93,05

94,21

95,12

93,28

Vitamin E (mg/kg) Temperature

a,b P<0.05; (vitamin E x temperature 0.01)

c b a a bb ab a ba

Antibody response (IgM and IgG) Macrophages (%)

Vitamin E effect on antibody response and macrophages (%) of broilers under heat stress (Arbor Acres, HS 23.9-38° C vs 23.9° C)

Higher Vitamin E dosage can improve antibody response (IgM and IgG) and macrophage %

Fonte: Niu Z.Y. et al., 2009

Vitamina A effect on immune response in heat-stressed (HS) layers (Hy-Line, 25 wks, 31.5°C)

Fonte: Lin H. et al., 2002

Humoral response

Vitamin A (IU/Kg)

anti

-New

Cast

le D

isease

Vir

us

Ab (

log2)

b

a

ab ab

35

40

45

50

55

3,000 6,000 9,000 12,000

T-C

ell P

rolife

rati

on (

%)

Vitamin A (IU/kg)

b

a

bc

c

a

bcbc

c

No Heat Stress

Heat Stress

Heat-stressed layers

after vaccination

require more vitamin A

for producing more

antiobodies

Measured at 7 and

14 dd post-

vaccinationa,b P<0.05

a,b,c P<0.05

Vitamina A and antibody response in heat-stressed broilers

Source: Niu Z.Y. et al., 2009

a,b,c P≤0.001; (vitamin A x temperature 0.001)

Vitamin A (IU/kg) Temperature

c

b

aa

b

Higher Vitamina A dosages can improve antibody

response (IgM e IgG) in heat-stressed broilers

Zinc and Vitamin A alleviate heat stress (34°C) effects in broilers

Source: Kucuk et al., 2003 Biological Trace Element Research

Control: diet containing 4,600 and 5,000 IU/kg vitamin A and 45 mg/kg Zn in grower and finisher phases respectively;

Zn: control diet + 30 mg Zn/kg diet;

Vit A: control diet + 4.5 mg (15,000 IU) retinol/kg diet;

Zn + Vit A: control diet + 30 mg Zn/kg diet + 4.5 mg retinol/kg diet.

Vitamin A and Zinc improved

performance in heat stressed

broilers alone and in combination

Blood parameters – emphasis MDA

and glucose – reflect the synergy

in absorption, metabolism and

effects of Vitamin A and Zimc

Gut Health

▪ Vitamin A

▪ Vitamin E

▪ Vitamin C

▪ Vitamin D3

▪ 25OHD3

▪ Vitamin B6

▪ Vitamin B2

UNSTIRRED WATER LAYER

Vitamin B6Glutammine-Aminoglycans

Thr-Cyst-Gly-Ser-ProMucin

Vitamin E

Vitamin A

ENTEROCYTE MEMBRANE

INTESTINAL PERMEABILITY

Reduced FAs peroxidation

VitaminD3/

25OHD3

Impact of vitamin D3 and 25OHD3 on intestinal morphology

Source: Chou S. H. et al., 2009

A, B, C and D are measurements at 14, 21, 28 and 35 days of age

Control: 3.000 IU/kg vitamin D3

25OHD3: 3.000 IU/kg vitamin D3 +

69 mg/kg (starter) and 34,5 mg/kg

(grower) of 25OHD3

25OHD3 significantly incresed villi lenght (duodenum and jejunum) and reduced crypt depth

The increased ratio villi lenght/crypt depth may suggest an enhanced rate of nutrient absorption

Faecalibacterium prausnitzii(prec. Fusobacterium prausnitzii)

▪ G(+), non-spore, anaerobic, difficult to cultivate even in anaerobiosis

▪ SCFAs producer, mostly butyric acid

▪ Reduces pro-inflammatory cytokines production (IFN-g e IL-12)

▪ Promotes production of anti-inflammatory (IL-10)

▪ In humans present in GI tract ≈5% of microbiota in feces

▪ ≈ 15%+: obesity

▪ <5% : Crohn disease; IBD

▪ Isolated in poultry, swine calves (and insects)

Riboflavine (Vitamin B2) acts as redox

mediator allowinf the strict anaerobe

F.prausnitzii to grow in intestinal epithelium

despite aerobic condition

Performance

▪ OVN

▪ Vitamin D3

▪ 25OHD3

Total Vitamin Intake

Avera

ge A

nim

al Resp

onse

Deficient

Sub-optimum

Optimum

Special Applications

NRC

• Below NRC levels

• Animals at risks of

developing clinical

deficiency signs and

disorders

• Above NRC levels

• Preventing clinical

deficiency signs and

disorders

• Inadequate to permit

optimum health and

productivity

• Offsetting factors

influencing vitamin

requirement

• Permitting optimum

health, productivity

and food quality and

nutritional value

• Above optimum levels

• Optimizing certain

attributes such as

immunity, meat quality,

bone health, etc.

Optimum Vitamin Nutrition Graph

Optimum Vitamin Nutrition (OVNTM) is about feeding

animals high quality vitamins in the right amounts and

ratios appropriate to their life stage and growing

conditions.

Optimum Vitamin Nutrition is a cost-effective range of

vitamin supplementation optimizing

• Animal Health and Welfare

• Performance

• Quality and Nutritional Value of Animal-origin Foods

The OVN™ Concept

Authors Year Country ROI

1. Coelho 2000 USA n.a.

2. Perez-Vendrell et al. (IRTA) 2002 Spain n.a.

3. Perez-Vendrell and Weber (IRTA) 2007 Spain n.a.

4. Zhang et al.* 2011 China 13:1

5. Araujo et al. 2012 Brazil 5:1

6. Iglesias et al. (Granja Tres Arroyos, Cobb) 2012 Argentina 2:1

7. Aviagen Product Dev. Center 2012 USA 3:1

8. NKP FARM 2012 Thailand n.a.

9. Aviforum 2013 Switzerland 2,8:1

10. INRA & ITAVI 2013 France 5,2:1

* Trial on layers

OVN Trials in Poultry: 2000 - 2013

0

1000

2000

0-21 days 0-40 days

Body Weight (g)

Industry OVN

0

0,5

1

1,5

0-21 days 0-40 days

FCR (g/g)

Industry OVN

a b a b

300

320

340

360

380

Breast weight (g)

Industry OVN

13,5

14

14,5

15

15,5

Breast yield (%)

Industry OVN

a b a b

Perez-Vendrell and Weber, 2007

Effect of Optimum Vitamin Nutrition on

Performance and Meat Yield of Broilers

25OHD3 vs Vitamin D3 effect on breast meat yield: Single trials results 2004 - 2012

25OHD3 improved 1% breast meat yield against vitamin D3

Source: Summary of experimental and field trials, 2004 - 2012

* Vitamin D3 vs HyD starter & grower; ** Vitamin D3 vs HyD starter; HyD starter vs HyD all feeds

1,0%

2,0% 2,0%

1,7%

0,6% 0,7%

1,3%

0,3%

0,7%0,6%

0,8%

0,5% 0,4% 0,4%

0,8%

0,3%

0,6%0,4% 0,3% 0,3% 0,4%

2,0%

3,9%

0,5%0,6%

1,2%

0,2%

1,0%

0,0%

0,5%

1,0%

1,5%

2,0%

2,5%

3,0%

3,5%

4,0%

4,5%

APSI,

2007

APSI,

2008

APSI,

2009

APSI,

Q1 2

010

Agri

Sta

ts,

2010 (

small…

Agri

Sta

ts,

2010 (

mediu

m…

Agri

Sta

ts,

2010 (

larg

e b

irds)

*

Saunders

-Bla

des

& K

orv

er,

…

Bra

y,

SF A

ust

in…

Bra

y,

SF A

ust

in…

Bra

y,

SF A

ust

in P

en s

tudy,…

Ark

ansa

s, 2

012

SPR,

2012

Pola

nd,

2006

IRTA,

Spain

, 2007

ILVO

, Belg

ium

, 2007

ILVO

, Belg

ium

, 2008

IRTA,

Spain

, 2009

Pola

nd,

2010

Spain

, 2005

Hungary

, 2006

Italy

, 2008 (

fem

ale

)

Italy

, 2008 (

male

mediu

m)

Italy

, 2008 (

male

larg

e)

Hungary

, 2010

Fra

nce,

2011

Italy

, 2012 (

male

larg

e)

Avera

ge

Bre

ast

Meat

Yie

ld %

im

pro

vem

ent

HyD

vs

Vit

. D

3

Page 71

72

Vitamin D3 and 25OHD3 studies on meat yield

▪ Birds:

▪ 1440 Cobb broilers ; 0-42days

▪ Treatments:

▪ 4 treatment groups

▪ 12 replications per treatment

▪ 30 broilers per pen

▪ Parameters measured:

▪ Serum 25OHD3 at 21d and 42d

▪ BW, FCR at 21d and 42d

▪ Breast meat yield at 42d

▪ 50 birds/treatment

▪ Expression of protein synthesis-related genes

(mTOR, S6K-1 and IGF-1 ) at 42d

Vignale et al., 2015Trial Details

Source: Vignale,K., J. of Nutrition, 2015

Effect Vitamin D3 and 25OHD3 on Breast Meat Yield (%)

P value= 0.02, SEM: 0.20

19.71 b 19.72 b

20.40 a

20.30 a

19,2

19,4

19,6

19,8

20

20,2

20,4

20,6

Control High D3 HyD 0-42 HyD 0-21

%

25OHD3significantly improved breast meat yield but not additional D3

Source: Vignale, K., J. of Nutrition, 2015

Effect Vitamin D3 and 25OHD3 Fractional Synthesis Rate (FSR) at 42 d

P value= 0.0406, SEM: 3.06

4.44 b 4.38 b

16.27 a

14.77 a

0,00

2,00

4,00

6,00

8,00

10,00

12,00

14,00

16,00

18,00


Fra

cti

onal sy

nth

esi

s ra

te a

t 42d %

/d

25OHD3significantly improved protein synthesis rate but not additional D3


0,00

0,50

1,00

1,50

2,00

2,50


IGF-1

*

Effect of vitamin D3 and 25OHD3 on the expression of protein synthesis-related genes at 42 d

0,00

0,50

1,00

1,50

2,00

2,50

3,00


mTOR

0,00

0,50

1,00

1,50

2,00


S6K-1

25OHD3 from 0 to 42 d significantly improved the

expression of protein synthesis related genes

* *

*


• mTOR (mammalian Target of Rapamycin) is a protein-

kinase important for cell growth and proliferation.

• Its activation is dependent on the effect of hormones or

specific nutrients (Aas)

• Both mTOR and S6K are genes connected with protein

synthesis

Meat and Egg Quality

▪ Vitamin A

▪ Vitamin E

▪ Vitamin C

Lipid Oxidation and Meat Quality

Lipid Oxidation = Deterioration of Meat Quality

Hydroperoxides,

Cholesterol oxides

Potentially

harmful

substances

Drip loss

Destruction

of Membranes

Formation of

Metmyoglobin

Colour

changes

Aldehydes,

Ketones

Oxidative rancidity,

bad odour / flavour

25 35

100

250

5 25 65 180

a-T

ocophero

l (m

g/g

pro

tein

)

Vitamin E (mg/kg feed)

Source: Sheehy et al., 1991

Deposition of alpha-Tocopherol in Thigh Muscle

Effect of Optimum Vitamin Nutrition on Vitamin E in breast meat (mg/100g)

Page 80

P vitamin level 0,0002; P density 0,3645; P interaction 0,5607

Vitamin E level: T1 and T3 18,9 mg/kg; T2 and T4 225 mg/kg

0,00

0,20

0,40

0,60

0,80

1,00

1,20

1,40

Control OVN Normal High

Vit

am

in E

in b

reast

meat

(mg/1

00g)

Group


Source: A.M. Perez-Vendrell et al., 2002

Effect of Optimum Vitamin Nutrition on breast lipid oxidation (TBARS, nmol/g)

Page 81

P vitamin level 0,0001; P density 0,0211; P interaction 0,0254

Vitamin E level: T1 and T3 18,9 mg/kg; T2 and T4 225 mg/kg

0,00

0,05

0,10

0,15

0,20

0,25

0,30

0,35

0,40

0,45


TBARS (

nm

ol/

g)

Group


Source: A.M. Perez-Vendrell et al., 2002

Vitamin fortification of eggs

Vitamin Potential increase & comments

Vitamin A 2-3 fold

Vitamin D3 6 to 10 fold attainable in 2-3 weeks

25OHD3 Naturally present in egg yolk of hens receiving vitamin D3

(0,5 to 8,1 mcg/100 g whole egg)

3-4 fold increase

Vitamin E 4 fold or more increase in 2 to 3 weeks (e.g. 200 IU/kg

feed from 119 to 606 IU/kg egg yolk)

Vitamin K Nearly 5-fold increase in hens fed 7,5 mg/kg feed

Folic acid 2-3 fold increase in 2 to 3 weeks

Biotin 3-5 fold; excess largely goes into albumen

Vitamin B12 3-4 fold; good response but high feed levels needed

Vitamin B2 2-3 fold but fairly refractory to accretion at high feed levels

Pantothenic acid 2-3 fold; plateaus quickly

Niacin 2-3 fold; limited data base

Vitamin B1 ≈2 fold; limited data base

Vitamin B6 ≈2 fold; very limited data base

▪ Vitamins are essential nutrients, required in very small amounts,

playing a pivotal role in all metabolic and physiological functions

Take Home Message

▪ Vitamins have a pretty limited feed cost impact but a great

responsibility in animal’s health and performance as well as on

quality of animal origin foods

▪ Vitamins, besides a direct impact on DWG and FCR, can modulate

several physiological aspects like immunity and stress and hence

providing indirect performance benefits e.g. reduced medications

Thank you for your attention

[email protected]

recommended vitamin levels for high performance broilers and layers - gilberto litta

Health & Medicine