Dietary phytonutrients as alternatives -to-antibiotics in ... · hormonal messengers. Hormesis: is a biological phenomenon whereby a beneficial effect (improved health, stress tolerance,

Dietary phytonutrients as alternatives-to-antibiotics in agricultural animals Mode of action in modulating cross-talks among immunity disease resistance and gut microbiotaL I L L E H O J H Y U NA N I M A L B I O S C I E N C E S A N D B I O T E C H N O L O G Y L A B O R A T O R YB E LT S V I L L E A G R I C U LT U R A L R E S E A R C H C E N T E R U S D E PA R T M E N T O F A G R I C U LT U R E A G R I C U LT U R A L R E S E A R C H S E R V I C EB E LT S V I L L E M D U S A

2nd International Symposium on Alternatives to Antibiotics OIE Paris France12-15 December 2016

2nd International Symposium on Alternatives to Antibiotics OIE Paris France12-15 December 2016

Consumers want a food system that assures food is safe affordable healthful and sustainable

Antibiotics as growth promotersMany different types of pathogens infect chickens during their lifetime Use of antibiotics is increasing

Signs of poor gut health

Dibner J 2010 Int Coccidiosis Conf 10 Guangdong China

AntibioticsDiet

DrugsDiseaseStress

Furness JB 2013 Nature Reviews GastroenterologyHepatology

Bischoff BMC Medicine 2011 924

Why do we want healthy gut

Structural Intestinal Integrity-Effective Digestion and

Absorption of Food

Balanced and Stable Intestinal Microbiota

Balanced Immune Response-Absence of GI Illness

Dysbiosis and subclinical enteritis

Inflammation and acute phase immune response

Oxidative and osmotic dysfunction

Gut barrier dysfunction

Impaired nutrient synthesis (vitamins minerals SCFA)impaired energy metabolism

Low-level effect of antibiotic growth promoters on body weight gains in broilers

(Oh ST and Lillehoj HS 2017)

Mechanisms of Antibiotic Growth Promoters (AGP)

1 Anti-microbial2 Reduce subclinical infections3 Reduce microbial use of nutrients4 Reduce growth-depressing microbial metabolites5 Enhance uptake of nutrients due to histologically thinner gut

in antibiotic-treated animals6 Reduce pro-inflammatory cytokineschemokines7 In germ-free mice antibiotics increase in adiposity8 In human newborns antibiotics is associated with obesity

AGP

Competitive exclusionEnteric pathogen

Mucosal protectionInnate and adaptive immunity

Energy efficiency

Maintenance

Beneficial effects of antibiotic alternatives

Affect GI mucosa

Stimulates enzyme secretion

Improve microbial flora

Nutrient digestion

GrowthReproduction

Report of the meeting of the OIE AD HOC Group on prioritization of diseases for which vaccines could reduce antimicrobial use in animals Paris April 21-23 2015

Bacitracin GP NE CO

Virginiamycin GP NE

Coccidiosis and Necrotic Enteritis

Lillehoj HS 2015 Avian Coccidiosis as a prototype intestinal diseases In Intestinal Health Key to maximize growth performance in livestock edited by Theo Niewold Wagenigen Academic The Netherlands Pp 71-116

Oh ST and Lillehoj 2016 The role of host genetic factors and host immunity in necrotic enteritis Avian Pathology 45313-316

Antibiotic alternatives-Mechanisms

Gadde U Lillehoj H 2017 Antibiotic Alternatives- A Review Animal Health Research Reviews In press

Essential oils key in fight against antibiotic useFeb 4 2016 1227 1

Essential oils are a key solution to an alternative for antibiotic growth promoters (AGPs) this was shown by research carried out by US food giant Cargill But what do essential oils achieve that other non-medicated feed additives donrsquot

Carbohydrate = 250 gramsProtein =100 gramsFat = 60 gramPhytonutrients =15 grams

Walsh et al 2007

ncbinlmnihgovPubmed

Function of Phytochemicals

1 Anti-oxidants carotenoids polyphenols2 Stimulates innate immunity3 Enhance adaptive (memory) immunity tumeric4 Stimulate enzymes Indole from cabbage5 Anti-bacterial allicin from garlic6 Promote beneficial bacterial growth7 Hormonal activity isoflavones from soy8 Physically block pathogen binding Proanthocyanidins9 Serve as signal transduction agents 10 Epigenetic regulation (Isoflavone tumeric)

Phytonutrientsphytochemicals Broad name for a wide variety of compounds produced by plants found in fruits vegetables beans grains and other plants

Furness 2016

Phytochemicals act on receptors on entero-endocrine cells to release hormonal messengers

Hormesis is a biological phenomenon whereby a beneficial effect (improved health stress tolerance growth or longevity) results from exposure to low doses of an agent that is otherwise toxic or lethal when given at higher doses

Son T Camandola S and Mattson M 2008 Hormetic dietary phytochemicals Neuromolecular Med 2008 10236-246

Outbreaks of Clostridial infections have been associated with certain drug use disruption in gut microbiota gut barrier failure bacterial translocation inflammation and acute death

Li Lillehoj et al 2010 Avian Pathology 39255 Li Lillehoj et al 2010 Avian Pathology 39247 Oh ST and Lillehoj 2016 The role of host genetic factors and host immunity in necrotic enteritis Avian Pathology In press

Host intestinal genes differentially affected by Necrotic enteritis in broiler chickens

19

Kim DK Lillehoj HS 2014 Transcriptional profiles of host-pathogen responses to necrotic enteritis and differential regulation of immune genes in two inbreed chicken lines showing disparate disease susceptibility PLoS ONE 9(12)e114960

Avian Coccidiosis

Coccidiosis are ubiquitous they are present wherever chickens arereared (traditional industrial label or organicbio farms) Nine speciesof Eimeria have been described in chicken

Balance of Th17 and Treg cells in Coccidiosis

Th17 and Treg cells have opposite actions on intestinal immunity

Treg cells suppress a variety of physiological and

pathological immune responses in the intestine

Th17 cells play a key role in intestinal inflammation

through induction of pro-inflammatory cytokines

Dendritic cells (APC) present the antigen to the T cells

Naiumlve CD4+ T cells differentiate into either Th17 or Treg

once in contact with dendritic cells

In gut immune systemhellip

IL-17 as a potential therapeutic target of coccidiosisIL-17

bull is pro-inflammatory cytokine produced by Th17 cells

bull induces various pro-inflammatory cytokines and

mediates inflammatory response

Administration of IL-17A neutralizing antibody induce

- Enhanced weight gain

- Reduced oocyst shedding

- Increased heterophil infiltration

- Reduction of cecal lesion

- Inhibition of E tenella schizont and merozoite development

Del Cacho et al 2014 Zhang et al 2013

Kim WH Jeong J Park AR Yim D Kim S Chang HH Yang SH Kim DH Lillehoj HS Min W 2014 Downregulation of chicken interleukin-17 receptor A during Eimeria infection Infect Immun 82(9)3845-54

Flow diagram for screening immunoactive phytochemicals

Disease challenge

Macrophage activation

Lymphocyte stimulation

Cytokine Tumor cytotoxicity Direct killing of pathogens

Select synergistic plants

Nutrigenomics

In vitro screening

In vivo feeding trial

Randomized Performance analysis in commercial broilers

In vitro and in vivo effects of purified phytochemicals on Eimeria and C perfringens

5 ppmcarvacrol

3 ppmcinnamaldehyde

2 ppmCapsicum oleor

23

246

625198

3126

48

156

31

Primary infectionE Maxima

Secondary infectionE Maxima

Up regulated Down regulated

Kim DK Lillehoj HS Lee S H Jang S and Bravo D 2010 High-Throughput Gene Expression Analysis of Intraepithelial Lymphocytes Following dietary Carvacrol Cinnamaldehyde or Capsicum Oleoresin Poultry Science 8968-81Kim DK Lillehoj HS Min WG Kim CH Hong YH and Lillehoj EP 2011 PlosOne V6e27712

NUTRIGENOMICS to identify 3 phytonutrient combination that reduced negative effects of avian coccidiosis

From black box to bold scienceLee Sung Hyen et al 2011 British J Nutrition

Phytochemical doses that we used were not cytotoxic to pathogens

Dietary effects of phytonutrients on coccidiosis

Lee S H Kim Duk Kyung Bravo D Lillehoj HS 2010 Journal Poultry Science 4789-95

Dietary Capsicum and turmeric oleoresins decreased inflammatory cytokinetranscripts and mitigated gut damage due to necrotic enteritis

Lee SH et al Lillehoj HS Bravo DM 2013 Dietary supplementation of young broiler chickens with Capsicum and turmeric oleoresins increases resistance to necrotic enteritis Br J Nutr110(5)840-7

IL-8

LITAF

TL1A

IL-17

IL-17F

Garlic effects in vitro

Kim DK Lillehoj HS Lee SH Lillehoj EP Bravo D 2013 Improved resistance to Eimeria acervulina infection in chickens due to dietary supplementation with garlic metabolites Br J Nutr 109(1)76-88

a

b

a

460

480

500

520

540

560

580

600

Non-infected Control Garlicon 40

Control E acervulina infected

Body

wei

ght

gai

n (

g)

BW gain

a

b

00

05

10

15

20

25

30

Control Garlicon 40

Oocy

sts

(X10

8 )

Oocysts

b

a

0

02

04

06

08

1

Control Garlicon 40

Ab r

esponse

to p

rofilin

(O

D

450n

m)

Ab-Profilin

In vivo dietary effects of Garlic

- Allium hookeri a member of the family Alliaceae is found in Ceylon Greece Southern China and India and has been used by locals to treat cough and cold and to heal burns and wounds(Sharma et al 2011)

- A hookeri contains an abundance of organo-sulphur compounds volatile sulphur compounds proteins prostaglandins fructans vitamins and polyphenols as well as Allicin

- Allicin and organic sulphur compounds are known to reduce cholesterol levels decrease the risk of heart attack and exert anti-inflammatory effects (Bae and Bae 2012 Kim et al 2012)

Allium hookeri

0

20

40

60

80

100

1000 500 250 125 625 1000 500 250 125 625 1000 500 250 125 625(μgmL)

Scav

engi

ng a

ctiv

ity (

)

Vit CControl Root Fermented rootLeaf

Free radical scavenging capacity

Interplay of nutrition gut microbiota and immunity

Dietary fermentable fiber and SCFAs can shape the immunological environment in the lung and influence the severity of allergic inflammation

Effect of phytochemicals on gut microbiota diversity in broilers

0

10

20

30

40

50

60

70

80

90

100

Variovorax

Geitlerinema

Calothrix

Clostridium

Selenihalanaerobacter

Streptococcus

Candidatus Arthromitus

Lactobacillus

Cobb Hubbard Ross

Kim JE Lillehoj HS Bravo D 2015 Dietary capsicum and curcuma longa oleoresins increase intestinal microbiome and necrotic enteritis in three commercial broiler breeds Res vet sci 102150-158

From plants to novel anti-parasite therapiesThe 2015 Nobel Prize winner in Medicine to Dr Youyou Tu concerning novel therapies against infections caused by Malaria protozoa

Malaria-mosquito-borne disease caused by single-cell parasites which invade red blood cells causing fever and in severe cases brain damage and death It claims more than 450000 lives

Strong evidence to suggest that the primary activator is an iron source Multi-faceted nature of cellular response to artemisinin in Plasmodia and tumor cells includes DNA damage which is dose dependent

-Resistance-Bioavailability-Cost and limitation-Mechanism of action