Phytogenics inpoultrynutrition;animportantsourceofantimicrobial,

anticoccidial,antioxidantandantiinflammatory compounds
LecturerDr.Ilias GIANNENAS
LaboratoryofNutrition,FacultyofVeterinaryMedicine,
AristotleUniversityofThessaloniki,Greece

FutureDynamicsinsustainableAnimalNutrition,
November6&7,2014,HartvanHollandconferencecentre,
Nijkerk,TheNetherlands

SCHOTHORSTFEEDRESEARCH
80YEARANNIVERSARYCONFERENCE

Presentationoutline
Introduction
Phytogenics
Plantbioactivecompoundsandhealthimplications

Antimicrobialeffect
Antiprotoozoal effects
Antioxidanteffect
Antiinaflammatory effect
Othereffects

Conclusions
Perspectivesforfutureresearch

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

Demands
Consumer

Producer

Good quality food

No residues

Optimum performance
Healthy crops

Antibiotic Growth Promoters

& Coccidiostats

Natural Alternatives

Growthpromotersasfeedadditives
The use of feed additives as a mechanism for manipulating growth

promotion in farm animals has been extensively investigated since 1950s

(Beeson and Perry, 1952)

Resistancetoantimicrobials!

Recently, animal production in many countries has faced restrictions in

the use of antibiotic growth promoters

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

ConsequencesofAGPsbanonguthealth
Dysbacteriosis / Wet Litter
problems
Decreased digestion
Reduced weight gain
Impaired feed conversion ratio

Ban on AGPs
2006

Necrotic enteritis

Coccidiosis

Intestinal necrosis

Intestinal necrosis
Mortality

Estimated cost
= 0.05$ / bird

Mortality

Alternativesforbacterialdiseases
Necroticenteritis significantlosses,cost,especiallyaftertheAGPban
Enteritis(E.coliinfection,Salmonellosis) mortality,bacteriatransfer
Omphalitis (Navelinfection) mortality,depressedgrowthrate
Chronicrespiratorydisease mortalitydepressedgrowthrate
AvianCampylobacterinfection bacteriatranfer,endangeredfood
chain
Footpaddisease depressedgrowthrate

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

Alternativesforprotozoaldiseases Coccidiosis
Coccidiosis is a significant endemic disease in poultry

3
Billions

The financial loss to

the poultry industry

worldwide circa
3 billion USD/year

2008
Cost mainly due to prophylactic or therapeutic infeed medications and also

as a result of the disease impact on birds health

(Williams,1999;Bozkurt etal,2013)

Thymol

Hippocrates,thefatherofmedicine,usedplantextracts
andprescribedperfumefumigations

Phytogenics

Thyme

Piperin
e

Black Pepper

Productsof plantorigin
o Botanicals
o Plantextracts and essentialoils
o Purecompounds
Lastdecadesseveralinvivo studieshavebeencarriedoutto
evaluatetheeffectivenessofessentialoilstomanipulate
growthperformanceandimprovenutrientutilizationon
poultry(Brenes andRoura,2011)

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

Phytogenics Activities&substances
Antimicrobial
Bactericide
Parasiticide
Fungicide
Insecticide
Antioxidant

Bioactive substances
Carvacrol
Thymol
Eugenol
Limonen
Vanilin
Cinnamaldehyde

Antiinflammatory

Tannins

Feedintakeincrease
Stimulatingdigestivesecretions

AntimicrobialmechanismofEOs
Thehydrophobicity ofEOsenablesthemto
partitioninthelipidsofthecellmembrane
andmitochondria,renderingthempermeable
andleadingtoleakageofcellcontents
DisintegratetheoutermembraneofGram
negativebacteriareleasing
lipopolysaccharides (Burt,2004)

Synergisticeffectprovidesanincreased
intensitycausedbythecombinationof
twoormoresubstancesonanorganism

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

Bacteriostatic/Bactericidal effectsofEOs
Rank

Chemical group

Component

Essential Oil

Phenols

Carvacrol, thymol, eugenol, gaiacol

Aldehyde

cinnamaldehyde

Inner bark of cinnamon tree

Monoterpnol C10

1-terpinne-4-ol

Tea tree

Aldehydes

Geranial, citronellal

Lemon grass

Cetones

Verbenone, menthone, carvone

Rosemary, aniseed, mint

Ether

Estragol, anethol

Basil, aniseed, star anise

Oxydes

Eucalyptol

Eucalyptus

Terpenes

Pinenes

Pines, firs

Oregano, thyme, clove, cinnamon leaves,

Knobloch et al., 1989; Franchomme et al., 1990; Dogna, 1990; Inouye et al., 2001; Friedman et al., 2002;
Hernander-Ochoa, 2005

Invivo studies
Onthisbasis,itislogicaltoconsideressentialoilsapplicationinvivo as

theintakeofessentialoilscouldreplaceantiobiotics

Vogt,earlyas1981,usedhersandspicesinpoultrynutrition
Essentialoils,preparationsofherbalextracts&blendswithpurephenolic

substancesshowedareductionofcolonyformingunitsofmanybacteria
suchasClostridiumspp

Herbalfeedsupplementscanaffectgastrointestinalmicroflora

compositionandpopulation.

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

Phytogenicsasgrowthpromoters
Dietaryuseof:
Oreganoplant
Oreganoextracts/essentialoils
Carvacrol,thymol

Initialresearchyielded:82papers
Suitableforthisreview:47papers
Parameter

N of
manuscripts

Maximum
deficit %

Maximum
benefit %

BW

-6.74

+23.00

BWG

N.S.

+14.11

Animal type

No of
manuscripts

Chicken broilers

26

Chicken hens

Turkeys

Quail

Rabbits

Pigs

Sows

-15.07

+9.20

Cows

13

-5.45

+17.19

Ewes

Mortality

N.S.

N.S.

Lambs

Egg

N.S.

N.S.

Milk

N.S.

+35.39

FI
FCR

Giannenas et al., 2013

Antiparasitic,protozoal&fungalmechanismsofEOs
Phenolsexhibitoocysticide activityasdisinfectantsinvivo &invitro

tests (Williams,1997)

Phenolsaretargetingagainstcellmembranesaffectingcellwall

structure,internalcellmembranesandenzymeactivities

Polyphenols (tannins)havedirecteffectstoparasites,butalsotothe

eggs

Activesubstancesreactwithsulfhydryl groups,whichare

indispensableforthefungalgrowth

Formate ofchargetransfercomplexeswithelectrondonorsinthe

funguscellthat
leadtoinhibitionofcelldivisionand
interferencewithcellmetabolism
inhibitionofthefungalcellwallsynthesizingenzymes

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

Currentinterventionstocontrolcoccidiosisinpoultry
Chemotherapeutics(antibiotics chemicals)
Controlofcoccidia hasbeengreatlydependentontheuseof

chemotherapeuticsagents
Ithasprovensuccessinmanypartsoftheworld,duetoitseaseofuseandthe

abilitytoprovideuniformtreatmentandprevention,however
Onsomeoccasions,mightbetoxictothebirds
Theconstantpressuretoreducethedependenceonantimicrobialsincludingthe
anticoccidial drugs,becauseof
Highcost
Publichealthconcernfordrugresiduesinpoultrymeat,eggs,foods
ResistancedevelopmenttodrugsbyEimeria species

Resistancetoanticoccidialdrugsworldwide
Approved drugs=Ionophores,synthetic chemicals,sulfonamides,mixed

products
Resistance theabilityofaparasitestraintosurviveand/ormultiply

despitetheadministrationandabsorptionofadrugindosesequaltoor
higherthanthoseusuallyrecommended
Developmentofresistancetoallanticoccidial drugs
WidespreadoccurrenceinUS,EU,SouthAmerica,SouthAfrica,China

(19742004)
Crossresistancetonewsubstanceswithsimilarmodeofaction!

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

Alternativecontrolofcoccidiosisinpoultry;
Phytogenics?
Plants/extracts/compounds

Scientific name

Challenge/beneficial effects/mode of action

Literature

Oregano aerial parts and

essential oil (containing
carvacrol and thymol)

Origanum
vulgare ssp.
hirtum

Eimeria mix (8 species): lesions, oocyst

shedding, FCR; E. tenella: body weight gain,
FCR, lesions, oocyst shedding

Batungbacal et al.
(2007), Giannenas et
al. (2003,2004)

Quinghao dried herb;

artemisinin; 1,8-cineol;
camphor; A. sieberi
petroleum ether extract of
aerial parts; A. afra
acetone/water extract from
aerial parts
Green tea leaves
China bark tree extract,
Quinine
Turmeric spice rhizome,
Curcumin

Artemisia annua, E. tenella: lesions, oocyst shedding; E.

A. sieberi., A.
acervulina: oocyst shedding (not E. maxima);
afra
Eimeria mix (E. tenella, E. maxima, E.
acervulina): FCR; induction of oxidative
stress.1,8-cineol and camphor: body weight
gain, lesions
Camellia sinensis
Cinchona
succirubra
Curcuma longa

E. maxima: oocyst shedding

E. tenella, E. meleagrimitis: sporozoite invasion
in vitro
E. maxima: lesions, body weight gain; E.
acervulina (not E. tenella); antioxidative

Allen et al.
(1997,1998), Allen &
Fetterer (2002), Arab
et al. (2006), Naidoo et
al. (2008)

Jang et al. (2007)

Christaki et al. (2004),
Fayer (1971)
Allen et al. (1998),
Allen & Fetterer(2002)

(1/2)
Bozkurt et al (2013)

Anticoccidialphytogenics
Plants/extracts/compounds

Challenge/beneficial effects/mode of action

Literature

Oriental plum; Japanese plum Prunus salicina

Scientific name

E. acervulina: body weight gain, oocyst

shedding, IFN- and IL-15 (mRNAs) of
intraepithelial lymphocytes, spleen cell
proliferation; phenolics and antioxidants

Lee et al. (2007)

Sugar cane extract

E. tenella: body weight gain, oocyst shedding,

lesions, antibody response
E. tenella: body weight gain, diarrhoea,
mortality, lesions, oocysts shedding
E. tenella: weight gain, mortality, bloody
diarrhoea
Eimeria mix (E. tenella, E. maxima, E.
acervulina): FCR, oocyst shedding

El-Abasy et al.
(2003)
Florou-Paneri et al.,
2004
Youn & Noh (2001)

E. tenella: body weight gain, mortality, lesions;

Eimeria mix (E. tenella, E. maxima, E.
acervulina): FCR

Naidoo et al. (2008),

Wang et al. (2008)

Olympus tea

Saccharum
officinarum
Sideritis scardica

Ku Shen root decoction

Sophora flavescens

Wild garlic acetone/water

extract from whole plant;
marasmine

Tulbaghia violacea

Grape seed proanthocyanidin

extract; ethanol/water extract
from pomace

Vitis vinifera
Artemisia, Thyme

(2/2)

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

Naidoo et al. (2008)

Bozkurt et al. (2013)

E. tenella in vitro ::: screening

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

10

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

11

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

12

b
c

c
d

E.tenella invivo::: challenge trial

Group

Code

Description

UU

untreated, uninfected

UI

untreated, infected

Las

Tenc

Inclusion
rate
(g/ton feed)

Lasalocid

75

Thymol

300

Thymol, matrix
encapsulated

140

2E+04 E. tenella
oocysts per bird at day
14 (crop intubation)

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

13

E.tenella invivo::: challenge trial

UU

UI

Las

Tenc

BW (g)

Day 35

1865a

1780bc

1842ab

1710c

1831ab

FI (g)

2986a

3206b

2931ac

2737 c

3046ab

FCR (FI/BWG)

1,641a

1,849c

1,632a

1,644a

1,706b

c
b
a

ab

FCR (FI/BWG)

BW (g)

ab

bc
c

UU

UI

Las

Tenc

E.tenella invivo::: challenge trial

UU

UI

Las

Tenc

LS d21 (0-4)

0,00a

2,67b

1,75b

2,38b

2,00b

LS d35 (0-4)

0,17a

2,17b

0,67a

2,00b

1,67b

2+

0,0ac

86,4b

19,4c

31,1b

DS d17-21
OPG reduction (%)
Mortality (birds/30)

Normal villae

Cocci affected villae

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

14

Effectsofcoccidia onintestinalvilli

Normal

Cocci affected

Theyneed96hourstorepair!
Dochickenshavethistime?

AntioxidantmechanismsofEOs
Phenolics reactwithfreeradicalsandreactiveoxygenspecieslike

tocopherols
Detoxifyandprotectcellandorgans
Spareeffectontocopherols likeascorbicacid
Prooxidant activityatcertainlevelslikeascorbicacid?
Antioxidantmechanismsnotyetfullyunderstood

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

15

Resultsonantioxidantactivity
1600

CONTROL
OR5
OR10
OR5-TC
OR10-C
C

MDA (ng/g)

1200

800

400

0
0

90
180
270
Incuba tion tim e (m in)

MDA levels in breast tissue submitted

to iron-induced lipid oxidation

AntiinflammatoryeffectsofEOs

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

16

Oreganosubstances
downregulated ileal IL6
andtonsilLITAF,IFN,
TLR4andIL10gene
expression
&exertedasignificant
antiinflammatoryeffect

E. tenella in vivo ::: screening

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

17

Phytogenics Digestibilityenhanceronchicken
Thymol,cinnamaldehyde oracommercialpreparationofessentialoil

components wereshowntoincreaseamylaseactivity intheintestinal

digesta offemalebroilers(Leeetal.,2003).
Increaselipaseactivity inthepancreas andintestinalwall withaplant

extract consistingofcapsaicin,cinnamaldehyde andcarvacrol withFCR

improvement inbroilers (Jamroz etal.,2005).
Dietarypiperine (pungentprincipleofblackpepper)stimulatedthe digestive

enzymesofpancreas andenhancedthedigestivecapacity andreduced the

gastrointestinalfeedtransittime (Srinivasan,2007).
Plantextracts fromamixtureoforegano,cinnamon andpepper orfrom

sage,thyme androsemary improved apparentwholetractandileal

digestibilityofnutrientsinbroilers(Hernandezetal.,2004).

Adverseeffectsofaromaticplants
Inaworkwithoreganogroundplant,althoughdietsupplementationat5.0

and7.5g/kgoffeedprotectedbirds

Oreganoat10g/kg,hadnonprotectiverolepossiblyduetoactionofits

phenolic constituentsthatinhighdosesmighthavetoxiceffects(Giannenas et
al.,2004)

Phenolic compoundsmayexerttheiractivityonthehostenterocytes,too

(WeberandDeBont,1996)

Carvacrol andthymol mighthavebreakingeffectontheupperlayerofmature

enterocytes ofmucosa,whicharealreadyaffectedbyintracellularpathogens

Hydrophobiccharacterofcarvacrol suggestsinteractionwithmembranes.

Whenconcentrationofcarvacrol increases,itisexpectedtoaccumulateand
interactinthephospholipid belayer,affectingmembranefluidity

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

18

Explanationsforinconsistentresults
Evidenceonbiologicalactivepropertiesofplantsisnotconsistentwiththe

expectationsarisingfromtradition

Majorityofethnopharmacologyreportsoriginatefromruminants,asthemain

livestockspeciesthatgenerateincomeinpoorcountries

Whensuchplantsaretestedinmonogastrics,partofthereportedvariation

maybeduetophysiologicaldifferencebetweenruminantandnonruminant
animals(Githiori etal.,2003)

Essentialoilsduetotheirpotentnatureshouldbeusedaslowaspossible

levelsinanimalnutrition

Theymayleadtofeedintakereductionorgastrointestinalmicroflora

disturbanceortoxicity

Explanationsforinconsistentresults(2)
Variationobservedintheexperimentalprotocolsandlevelof

supplementations
Variouseffectsongutmicroflora andintestinalprotozoabutalsoon

animalmetabolism
Variable composition.For example,concentration of two predominant

components of thyme essential oils,i.e.thymol and carvacrol have been

reported to range from 3%to 60%of total oils
Observedlackofgrowthpromotingeffectinsometrialsmayrelateto

compositionofbasaldietand/orenvironmentalconditions

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

19

SafetyIssues?
FoodgradelistedasGRAS

(GenerallyRecognizedAsSafe)
AppearonEURegisterofFeedAdditives

(Reg.1831/2003)
Residueshavetobemonitored

ShareofphytogenicsinEUmarket

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

20

Productionofphytogenics
Authentification of raw materials
Chemical standardization
Process chemistry: scale up
In vitro
studies

In vivo
studies
Field Trials

Quality
control

GMP production and marketing

Futureperspectives
Newinvestigationsarewarrantedonthefollowingtopics:
mechanismsofaction forpromisingcompoundsincombinations
(e.g.EOswithhealthbenefits,+probiotics,+othercompoundse.g.organicacids)
bioavailabilityandpharmacokinetics ofEOs+otherfeedadditivese.g.enzymes
zootechnical andbiologicaleffectsofcombinationsofalternatives basedonthe
factthatGITdisordersposthatchingdisplayhighlycomplexaetiologyand
mechanisms,interactionwithimmunity
negativeinteractions betweenfeedcomponentsandaddedalternativesORamong
alternativesmustbeunderstood

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

21

NovelSearch
Numerousnativeherbsarerichinphenolic compounds
However,multidisciplinaryresearchisnecessarywiththeinteractionsof

researchInstitutesand,theFeedAdditiveindustry,theFeedindustryand
thePoultryindustryinordernovelsubstancestobetested!
Progresscanbeachievedbyconsideringthevariousaspectsofmedicinal

plantstogetherandthusexploittheircomplementarity

Emphasisandadditionalongoingresearchfor
performance,
immunity
behaviouralobservations

MainReferences
Aromatic plantsasasourceofbioactivecompounds:E.Christaki,E.Bonos,I.
Giannenas,P.FlorouPaneri,Agriculture,2012
An update on approaches to control coccidia in poultry using botanical extracts: M.
Bozkurt, I. Giannenas, K. Kucukyilmaz, E. Christaki, P. FlorouPaneri, British Poultry
Science, 2013
Essential oils and their applications in animal nutrition Invited Review: I. Giannenas,
E. Bonos, E. Christaki, P. FlorouPaneri, Medicinal and Aromatic plants, 2013
Oregano supplementation in farm animal diets: effects on productive parameter: .
Giannenas, . Bonos, E. Christaki, P. FlorouPaneri. 17th Congress of the European
Society of Veterinary and Comparative Nutrition, Ghent, Belgium, 2013.

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

22

Thankyouforyourattention!

Thessaloniki, Greece

Future Dynamicsinsustainable Animal Nutrition,Nov7,2014

23