Phytonutrients as alternative feeding strategy to improve

performance of cattle without using an antibiotics

Sergio Calsamiglia

Dpt. Ciencia Animal y de los Alimentos

Universidad Autónoma de Barcelona

Opportunities for improving feedefficiency

VandeHaar et al., 2014

Objective: Energy retention

50

70

90

110

130

Acetate Propionate Butyrate

Efff

icie

ncy;

%

Gràfic1

Acetate

Propionate

Butyrate

Effficiency; %

62

108

78

Sheet1

Acetate62

Propionate108

Butyrate78

Fiber Starch

Pyruvate

Acetate

Methane

Propionate

Lactate

Butyrate

Glucose+4 H+

+4 H+

-8 H+

-4 H+

-2 H+

-2 H+

Energy efficiency in rumen fermentation

Monensin⇓ Gram +⇑ Gram -

⇓ Catabolism Pep & AA

⇓ NH3

⇑ Propionate⇓ Acetate⇓ Butyrate

Effects of monensin

Effects of monensin

-4

-3

-2

-1

0

1

2

3

DMI Mik yield DMI ADG

Varia

tion,

% C

ontr

ol

Dairy; Duffield et al., 2008 Beef; Duffield et al., 2012

Objectives

Search for alternatives:

Tradicional approach: a “ionophore-like” product

“Unexpected targets”

Plant Extracts

Products of the secondarymetabolism.

Small size, high diversity

Clasified as GRAS (GenerallyRecognised as Safe) (D-1999/217/CE; FDA, 2004). O

Timol

O

1-8-cineole

Carvacrol

Terpinen-4-ol

OH

OH

Carvone

Limonene

OH

O

H

OH

Cinamaldehído

Anetol

Eugenol

O

O

Glucose Pentose

Phospho-enol pyruvate

3 Acetyl- CoA

Mevalonate

DOXP

Isopentenyl Pyrophosphate

MonoterpenesTetraterpenesSterolsTriterpenes

Erythrose 4 phosphate

Shikimic acid

Chorismate

Cysteine

PhenylalanineTyrosine

Cinamic acidCoumaric acid

Anethole Eugenol Cinnamaldehyde

IsoflavonesTaninsLigninsPhenylpropanes

ThymolCarvacrol

Capsaicin

Allylsulfides

Terpenes

SaponinsSarsaponins

Mechanism of action

Ultee et al., 2002

Modulation of microbial population

Patra and Yu 2012

Phytonutrient selection process

500 molecules / extracts

100 Tested in vitro - batch

10 Tested in vitro Continuous Culture

5 Tested in vivo

CTR MON CIN

LPep N 6.8 5.4 6.4

SPep + AA N 1.9 4.4† 2.5

Ammonia N 19.2 13.0* 16.0 †

* P < 0.05 † P < 0.10

Nitrogen metabolism

Busquet et al., 2005 (J. Dairy Sci.)

1* 2 3 4 5 6 7 8 9 10 11 12 13 14

Fluore

scence

Time (seconds)

0123456789

1011

35 37 39 41 43 45 47 49 51 53 55

CTR1 P2 restrikcija HaeIII CIN1 P2 restrikcija HaeIII PCR GAR1 P2 neredcenCTR 1 (P2) HaeIII digestion CIN 1 (P2) HaeIII digestion GAR 1 (P2) undigested PCR product

CTR CIN

P. rP. rP. b

P. b

Mechanism of Action: (Prevotella)

(Ferme et al, 2004)

Nitrogen metabolism

CTR FEN CAD TEA DIL GIN CLO

LPep N 3.8 4.5 6.1 4.3 5.0 4.8 6.9

SPep + AA N 5.2 5.0 5.0 4.5 4.7 4.0 2.8

Ammonia N 7.5 8.7 7.4 8.0 8.5 7.1 6.4

*

†

* P < 0.05 † P < 0.10 Busquet et al., 2005 (Anim. Feed Sci. Technol.)

DietaryProteinProtein

PeptidesAA

NH3

Urea

Microbial Protein

XX EUGCIN

EO

Essential Oils: Protein metabolism

0

1.25

2.5

CTR MON CIN

Busquet et al., 2005 (J. Dairy Sci.)

Efficiency of energy utilization(Acetate to propionate ratio)

a

b

ab (P

Efficiency of energy utilization(Butyrate)

0

5

10

15

CTR MON CIN

mol

/100

mol

Busquet et al., 2005a (J. Dairy Sci.)

bb

ab (P

Fiber Starch

Pyruvate

Acetate

Methane

Propionate

Lactate

Butyrate

Glucose+4 H+

+4 H+

-8 H+

-4 H+

-2 H+

-2 H+

Hypothesis: Inhibition of methanogenesis

XTX

Effects of plant extracts

0

1

2

3

Milk yield, L/d; Bravo et al., 2009 ADG, kg/d; Bravo et al., 2009

% v

aria

tion

from

Con

trol

Dairy Beef

M (2.3%)M (2.5%)

Summary

Plant extracts modulate microbial populationin the rumen

Changes in the fermentation profile aresimilar to ionophores in direction and size, although a different mechanism of actionprobably takes place

Production performance in dairy (milk yield) and beef (ADG) is improved with a similar magnitude to that observed with Monensin

The unexpected targets

The ban on the use of antibiotics has open the door to new opportunities

Accidental findings are often the start of new discoveries

Feed efficiency is not only what happens in the rumen, but it is also affected by:

Lower digestive tract

Metabolism

Immunity and defense in the smallintestine

Goblet cellssecreting mucine (Muc2)

Mucine is a major constituant of inner mucus layer

Outer mucus layer & microbiota

Jamroz et al., 2006

Defense and immunity (swine)

0%

50%

100%

150%

uninfected CT infected CT infected + capsicum

MUC2 expression rel. control in ileal mucosa (fold change)

*

0%20%40%60%80%

100%120%140%

uninfected CT infected CT infected + capsicum

TLR4 expression rel. control in ileal mucosa (fold change)

*

↘ Host sensitivity↗ physical barrier

Liu et al. 2014, J. Anim. Sci

Piglets infected (challenged) with E. Coli without or with Capsicum oleoresin

Improved immunity in cows

20

30

40

50

60

0 200 400 600 800 1000 1200

Neutrophils Lymphocites

Capsicum, mg/d

% T

otal

cel

ls

Oh et al., 2015

Capsicum and milk production

46.6

49.548.3

47.3

40

42

44

46

48

50

52

Control C250 C500 C1000

Milk

, kg/

d

+6.2%

Oh et al., 2015

Capsicum and immunity:Total treatments (Calves)

56

48

44

48

52

56

60

Control XTRACT

Inci

denc

e, %

Capsicum and immunity: Treatments/sick calf

2.21

1.67

0

1

2

3

Control XTRACT

Trea

tmen

ts, n

Capsicum and immunity:Respiratory diseases (feedlot)

15.7

9.9

0

3

6

9

12

15

18

Monensin XTRACT

Inci

denc

e, %

Incidence of rumen lessions(feedlot)

73

24

0

10

20

30

40

50

60

70

80

Monensin XTRACT

Inci

denc

e, %

Hagg et al., 2013

Summary of Capsicum (1)Changes in the immune profile is consistent with improved immunity

Reduction in the incidence of diseasesobserved in calves and feedlot cattle

Difficult to justify a 6% increase in milkproduction (2.9 L/d) due to improved imunityin dairy

40

42

44

46

48

50

Control C250 C500 C1000

Milk

, kg/

d

Capsicm and milk prodution

C on tr

o l

N ex u

lin

C on tr

o l

N ex u

lin

C on tr

o l

N ex u

lin

0

1 5

3 0

4 5

Mil

k y

ield

(k

g/

d)

< 1 0 0 D IM

1 0 0 -2 0 0 D IM

> 2 0 0 D IM

* * *

D a y o f tr ia l

Mil

k y

ield

(k

g/

d)

P re -

t ria

l 5 7 9 1 2 1 5 1 9 2 2 2 5 2 8 3 1 3 4 3 7 4 0 4 2

3 0

3 5

4 0

4 5

5 0

C o n t r o l

N e x u lin

< 100 DIM

40,7 vs 44,4

Wall and Bravo, 2016

ControlCapsicum

Capsicum and milk production

W e e k o f la c ta tio n

Milk

yie

ld (

kg/d

)

1 2 3 4 5 6 7 8 9 1 0 1 1 1 2 1 3 1 4 1 51 5

2 0

2 5

3 0

C o n t r o l

N e x u lin

30,1 vs 32,1

Stelwagen et al., 2016

Control

Capsicum

XTRACT and milk yield(Heat stress)

39.4

42

36

39

42

45

Control XTRACT

Milk

, L/d

Blanck et al., 2016

The “other effect” of Capsicum

2.33.0

6.7 6.2 6.6

9.1

13.0

0

3

6

9

12

15

Monensin C+E C+E+C CAP CAP CAP bST

Varia

tion,

% C

ontr

ol

Duffield et al. 2008

Bravo, 2009 Ohet al., 2015

Blanket al., 2014 Stelwagenet al., 2016

Wall al., 2016

Bauman et al., 1999

Capsicum and insulin

0 2 0 4 0 6 0 8 00

2 0

4 0

6 0

8 0

T im e p o s t G T T (m in )

Insu

lin

,µ

IU/m

L

C o n t r o l

N E X U L IN ®

(Penn State U., Unpublished)

ControlCapsicum

Glucose metabolism

Adipose tissue

Glucose

Mammary gland

INSULINR

GlucoseRR

R

R

R

Glucose and early lactation (and bST)

Adipose tissue

Glucose

Mammary gland

INSULINRGlucose

Glucose and early lactation

Adipose tissue

Glucose

Mammary gland

INSULIN

Glucose

RRR

R

R

R

SummaryCapsicum enhances milk production (~6-9%)

The chagnes in insulin effects are parallel to those observed in early lactation or bST and may explain the mechanism of action thatjustified the increased production

Although capsicum increases feed intake, results suggest that this may not be thecause of increased production, rather theconsequence

“Playing with glucose”Sucram is a sweetner that stimulates sweetreceptorsIt may be used to increase DM intake

S

100%

133%

C

S

C

1, Texas A&M; 2, Texas Tech U

Receiving calves (190 kg)

CTR SUCRAMDMI, kg/d1,2 4,45 4,77 +7,2%ADG, g/d1,2 1242 1333 +11,4%ADG/DMI1,2 0,257 0,266 +5,9%

Pulls, %1 59,7 58,0 -2,9%Re-Pulls, %1 52,6 41,0 -22,0%

How can we explain theimproved efficiency?

Glucose absorption

Glu

Na

Glu

Glu

Na

K

ATP

Small Intestine

Blood

(Shirazi-Beechey, U Leeds, UK)

Regulation of glucose transport

Na ATP

AAAAAcAMPRE

ATPcAMPVPAC1

AC

SGLT1Sensor

T1R2/T1R3

Gustducin

Increased Ca2+

+++

Na+

GLP-2

Enteric neuron

GLP-2RVIP/PACAP

(Shirazi-Beechey, U Leeds, UK)

Changes in glucose transportersinduces by Sucram in calves

F

SGLT1

E SGLT1

Control Sucram

Moran et al., 2014

Results in calves: glucosetransporters

x2

x58

Gràfic1

Milk fedMilk fed

WeanedWeaned

Control

Sucram

pmol/s/mg prot

115

234

1.3

76

Sheet1

Milk fedWeaned

Control1151.3

Sucram23476

SGLT1 Gustducin

Increased Ca2+

+++TRPM?

Na+

GLP-2

Circulation Vagal neuron

Spinal afferent fibres

Enteric neuron

GLP-1

Insulin

GLP-1 and insulin release

(Shirazi-Beechey, U of Leeds)

Sucram and insulin realease

Ponce et al., 2007

Insu

lin, n

g/m

L

Time, minutes

Gràfic1

00

1515

3030

4545

6060

7575

9090

105105

120120

135135

150150

165165

180180

Control

Sucram

1

2.1

2

2.5

1.5

2.8

1.9

2.7

1.9

3.7

1.3

3.2

1.9

2.6

1.8

2.7

1.4

3.5

1.9

2.9

1.8

3

1.4

3.8

1.7

3.5

Full1

ControlSucram

012.1

1522.5

301.52.8

451.92.7

601.93.7

751.33.2

901.92.6

1051.82.7

1201.43.5

1351.92.9

1501.83

1651.43.8

1801.73.5

Per redimensionar l'interval de dades del gràfic, arrossegueu l'angle inferior dret de l'interval.

Insulin in dairy and beef

Insulin release may reduce body fat mobilization and help in the control of ketosis (energy balance) post partum

Insulin increases growth of muscular and adipose tissue

The effect on intramuscular fat is highercompared with external fat

The effects on marbling is higher early in the growing period

ConclusionAntibiotic growth promotants have been usedsuccessfully in animal nutrition

The ban on the use of antibiotic growthpromotants (in the EU and likely in other coutries) has stimulated the research on alternativeapproaches

The need for such alternatives has also openednew oppotunities to identify “unexpected targets”

ImmunityGlucose metabolism

ConclusionPhytonutrients (namely Cinnamaldehydeand Eugenol) modify rumen microbialpopulation, the fermentation profile and dairy and beef performance, in a directionand size similar to some AGP.Capsicum has shown immune enhancingcapabilitiesCapsicum and Sucram may enhanceproduction performance of dairy and beefthrough the modification of glucosemetabolism

Phytonutrients as alternative feeding strategy to improve performance of cattle without using an antibioticsOpportunities for improving feed efficiencyObjective: Energy retentionEnergy efficiency in rumen fermentationEffects of monensinEffects of monensinObjectivesPlant ExtractsSlide Number 9Mechanism of actionModulation of microbial populationPhytonutrient selection processNitrogen metabolismSlide Number 14Nitrogen metabolismEssential Oils: Protein metabolism�Slide Number 17Efficiency of energy utilization�(Butyrate)Slide Number 19Effects of plant extractsSummaryThe unexpected targetsImmunity and defense in the small intestineDefense and immunity (swine)Improved immunity in cowsCapsicum and milk productionCapsicum and immunity:�Total treatments (Calves)Capsicum and immunity: �Treatments/sick calfCapsicum and immunity:�Respiratory diseases (feedlot)Incidence of rumen lessions (feedlot)Summary of Capsicum (1)Capsicm and milk prodution Capsicum and milk productionXTRACT and milk yield�(Heat stress)The “other effect” of CapsicumCapsicum and insulinGlucose metabolismGlucose and early lactation (and bST)Glucose and early lactationSummary“Playing with glucose”Receiving calves (190 kg)Glucose absorptionRegulation of glucose transportChanges in glucose transporters induces by Sucram in calvesResults in calves: glucose transportersSlide Number 47Sucram and insulin realeaseInsulin in dairy and beefConclusionConclusion