67th EAAP Meeting Belfast 2016 1 · This project has received funding from the European Union‘s Seventh Framework Programme for research, technological development and demonstration

This project has received funding from the European Union‘s Seventh Framework Programme for

research, technological development and demonstration under grant agreement No. 311794.

1 67th EAAP Meeting Belfast 2016



2

SYNBIOTIC

ECO FCE: IN OVO MANIPULATION TO INDUCE LONG TERM EFFECT ON MORPHOLOGY OF CHICKEN GUT

www.Nutraingredients.com

67th EAAP Meeting Belfast 2016



IN OVO TECHNOLOGY TO DELIVER SYNBIOTICS-MOVIE

67th EAAP Meeting Belfast 2016 3

HIGH TOLERANCE OF EMBRYOS FOR TREATMENT

HIGH HATCHABILITY RATES AFTER INJECTION

This prototype is a property of enterprise Drobex-Agro Sp. z o.o., Poland, according to grant agreement THRIVE RITE, No. 315198 (2012-2015)



WHAT ARE SYNBIOTICS?


Synbiotic = + prebiotic

substance present in or introduced into the food to stimulate the growth of gut flora

selected bacterial or yeast cultures which task is beneficial effect in the gastrointestinal tract

Synbiotic 1–

Lb. salivarius IBB3154 +Bi2tos, Clasado Ltd.

Synbiotic 2 – Lactobacillus plantarum IBB3036 + lupin RFOs

probiotic



Our Bioscreen result show a perfect match of Bi2tos and probiotic against glucose reference


SYN1 Lb. salivarius 3154 + Bi2tos SYN2 Lb. plantarum 3036 + RFO

Measured using a broadband filter (420nm-580nm wavelength range )



completely developed and highly vascularized

allantochorion efficient transport route from the air

cell to the blood [Romanoff, 1960]

Immersed in amniotic fluid contact with GI tract delivery in ovo

ensures reaching GI tract [Jankowski et al., 2012]

Why in ovo technology?

6

To ensure the best protection for the newly hatched individual, the external supplementation should be given as early as possible [Madej and Bednarczyk, 2016]

Broiler chicken life span

HATCH

1 day

SLAUGHTER

-12 day

Incubation of fertilized eggs

IN OVO INJECTION

42 day

www.jakesfeed.com www.hitechhatchfresh.com

Time from injection to hatch: promote

microflora development

Nenonatal period: stimulation of good health condition without antibiotic treatment [Madej et al., 2015]





EXPERIMENTAL METHODS

www.cartoonnetworkasia.com



EXPERIMENTAL REARING IN PRODUCTION CONDITIONS

8

Performance ( including the FCR) Seperate pens for sampling

Treatment Replicates

(pens)

No. of chickens

males/pen

Replicates

(pens)

No. of

chickens

males/pen

The sume of

experimental

chickens

SYN1 8 75 8 10 680

SYN2 8 75 8 10 680

CONTROL 8 75 8 10 680

+ 8000 * 2040


*„chicken house flock”: 8 000 chickens (not sexed) to fill the space of the chicken house; randomised, group housed experiment

in ovo injection: 5 850 Cobb 500FF eggs SYN1 Lb. salivarius 3154 + Bi2tos, Clasado Ltd. SYN2 Lb. plantarum 3036 + RFO, in house developed




EXPERIMENTAL RESULTS




FINAL BODY WEIGHT & FEED CONSUMPTION

10

SYN1 Lb. salivarius 3154 + Bi2tos, Clasado Ltd. SYN2 Lb. plantarum 3036 + RFO, in house developed

0

1000

2000

3000

CONTROL SYN1 SYN2Bo

dy

Wei

ght

Gai

n [

g]

BWG [1-41d]

0

0,2

0,4

0,6

0,8

1

1,2

1,4

1,6

1,8

CONTROL SYN1 SYN2

Feed

Co

nsu

mp

tio

n E

ffic

ien

cy

[g/g

]

FCE [1-41d]


0

1000

2000

3000

4000

CONTROL SYN1 SYN2

Feed

Inta

ke [

g]

FI[g]



MORTALITY

11

Very low, <2% mortality in ferm conditions

FCE did not significantly increase after synbiotics. In previous studies, an increase in prebiotic treated groups was observed

0

0,2

0,4

0,6

0,8

1

1,2

1,4

1,6

1,8

2

CONTROL SYN1 SYN2

Mo

rtal

ity

[%]




Weight and lenght of small gut at 42d


0

20

40

60

80

100

120

140

Control SYN1 SYN2

We

igh

t [g

]

020406080

100120140160180200220

Control SYN1 SYN2

Len

ght

[cm

]

b a ab

AB

B

A

A,B,C- P<0.01, a,b,c- P <0.05



Villi height, widht and crypt depth


0

500

1000

1500

2000

duodenum jejunum ileum

He

igh

t o

f vi

lli [

µm

]

0

50

100

150

200

250


Wid

th o

f vi

lli [

µm

]

0

50

100

150

200


De

pth

of

cryp

ts [

µm

]

CONTROL SYN1 SYN2

B A

AB

B B

A

AB A B

A

B B

B A A

A B B

A,B,C- P<0.01



14

Histomorphology- villi surface Day 42

CONTROL SYN1 SYN2

0

0,2

0,4

0,6

0,8

1

1,2

1,4

1,6

1,8

Duodenum Jejunum Ileum

Vill

i su

rfac

e m

ln µ

m2

A,B- P<0.01


A B AB

SYNBIOTICS positively affected surface of the villi.

SYN1 and SYN2 increased the villi surface in duodenum.




0

200

400

600

800

1000

1200

1400

Ne

utr

al g

ob

let

cells

/m

m2

/vill

us

15

Number of neutral goblet cells/mm2 villus

SYN1 –shielding effect from goblet cells + low mortality is a good effect during 1st period of rearing

SYN1 and SYN2 – did not affect neutral goblet cells at the end of rearing. Maintained good status. Displayed DIFFERENT MODE OF ACTION.

CONTROL SYN1 SYN2


B

A

C

b

a

c

a a b b a a a b b

1d 7d 42d

A,B,C- P<0.01, a,b,c- P <0.05



SUMMARY





17

LONG TERM EFFECTS ON GUT


surface of villi in dudodenum

surface of villi in small intestine (dudodenum & jejunum)- NS


goblet cells profile

depth of crypts depth of crypts

versus Control

lenght of dudodenum & jejunum lenght of intestine

goblet cells profile



MAJOR CONCLUSIONS ON GUT MICROSTRUCTURE


SYN1 and SYN2 positively affect (increase) the surface area of the absorbent villi of the small intestine throughout the rearing period

SYN1 and SYN2 positively affected number of goblet cells which might be cause of observed lower mortality

Different synbiotics show differential effect on the microstructure of individual sections [duodenum, jejunum, ileum] of small intestine

A single in ovo treatment with a low dose of synbiotic beneficially affects the gut microstructure (reflecting physiological state of a gut) at day 42

Synbiotics stimulated changes in duodenum and jejunum- 2 segments responsible for the most intensive absorption of nutrients in chicken gut

19

Inulin + L.Lactis ssp. lactis

Bi2tos + L.Lactis ssp. cremoris

RFO + L.Lactis ssp. lactis

RFO + L.Lactis ssp. cremoris

RFO+ L.plantarum

Bi2tos + L.salivarius

Duolac RFO Inulin Bi2tos DiNovo

BW [1-3]

BW [1-6]

FINAL BW

Breast muscle

Microvilli lenght

Microvilli surface

Bursa fabricius to spleen

IgG (Y) concentr.

Genes cecal tonsils

Genes spleen

Genes intest.

Peripheral lymphocytes B

Intestine Lymphocytes B

Ceacal tonsils lymphocytes

Pancreas potential



MAJOR IMPLICATIONS-FUTURE R&D


To reduce the production costs of healthy poultry meat, without use of antibiotics

Technical alignment with HATCHERY production line (the compatible elements)

Improved FCE Focus on meat quality (white stripping

reduction, beneficial fatty acid composition)

challenge tests –stress conditions + infections (Salmonella, Campylobacter)

To lower coccidia oocysts in young chickens.

In ovo & synbiotics



21 67th EAAP Meeting Belfast 2016

This scientific work is co-financed from funds for science of the Polish Ministry of Science and Education allocated to an international project ECO FCE in the years 2013-2017

UTP TEAMS:

UTP Leader: Prof. Marek Bednarczyk

THE WHOLE TRANSCIRPTOME ANALYSIS Prof.Maria Siwek Anna Sławińska, PhD Aleksandra Dunisławska, MSc- PhD student

PHYSIOLOGIC PARAMETERS AFTER SYNBIOTICS IN OVO Prof. Paweł Maćkowiak Prof. Leszek Nogowski Ewa Pruszyńska-Oszmałek, PhD Paweł Kołodziejski, PhD

HISTOLOGICAL PARAMETERS AFTER SYNBIOTICS IN OVO Joanna Bogucka, PhD Prof. Gabriela Elminowska-Wenda Agata Dankowiakowska, PhD Adriana Sobolewska, PhD Agata Szczerba, MSc- PhD Student

BIOCHEMICAL PARAMETERS AFTER SYNBIOTICS IN OVO Prof. Bogdan Janicki Mateusz Buzała, PhD Ola Roślewska, PhD Dorota Cygan-Szczgielniak

EXPERIMENTAL REARING, FISH BACTERIOLOGY Prof. Damian Józefiak

MEAT QUALITY Professor Giuseppe Maiorano Siria Tavaniello, PhD Cinzia Abiuso, PhD

UP POZNAN PARTNER:

GENETICS & STATISTICS Prof.Tomasz Szwaczkowski

Documents

67th EAAP Meeting Belfast 2016 1 · This project has received funding from the European Union‘s Seventh Framework Programme for research, technological development and demonstration