The Effect of Dietary Nitrate on Enteric Methane Emissions ... - J. Newbold.pdf · Jolien Veneman,...

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The Effect of Dietary Nitrate on

Enteric Methane Emissions and

Methaemoglobin in Ruminants:

a Meta-Analysis

www.cargill.com/feed

Jolien Veneman, John Newbold,

Hink Perdok, Jamie Newbold

February 2016

Introduction

NITRATE

• Nitrate acts as alternative hydrogen sink when being reduced to

ammonia (Ungerfeld and Kohn, 2006)

NO3- + H2 → NO2

- + H2O

NO2- + 3∙H2 + 2∙H+ → NH4

+ + 2∙H2O

• Both reactions s have a greater negative ΔG value than methane

formation, through the reduction of CO2, therefore nitrate reduction

would be favoured over methanogenesis as a hydrogen sink

• If nitrate is completely reduced to ammonia then 100 g nitrate could

‘compete’ with 25.8 g of methane (stoichiometric potential)

• When nitrite forms, and is not quickly reduced to ammonia, it can be

absorbed across the rumen wall to bind to Heamoglobin, forming

Metheamoglobin (MetHb). This can lead to reduced performance or in

extreme situations death at high levels of MetHb

2

Introduction

META- ANALYSIS

• Several newly published studies on nitrate supplementation and methane emissions

• Not included in Lee and Beauchemin (Dec., 2014) 11 comparisions.

Aim

• Study the effect of nitrate supplementation on methane emissions (g/kg DMI) Calculate efficacy of nitrate

• The effect of nitrate on MetHb%

• Which moderators influence this effect

3

Materials and Methods;Data selection

STUDIES

• 18 papers

Total comparisons between a control and nitrate treatment

• 27 for Methane yield, 21 for MetHb%

Inclusion criteria;

• Within each experiment, control and nitrate diets contained equal concentrations of crude protein.

• Methane yield was expressed as methane emissions per kg dry matter intake (DMI) and MetHb as % of total haemoglobin.

4

Materials and methods; analysisMETAFOR PACKAGE IN R (VIECHTBOUWER, 2010)

• Mean difference (MD) (Treatment – Control) can be used when units (and range) are the same

• Using Random effects model to test overall effect of nitrate;

• θi = µ + ui

• Mixed effects model to test effect of moderators;

• θi = β0 + β1 x1p + ……. + βp xip + ui,

• Each moderator was first tested individually and included if P<0.10 to construct final model

• Animal type (beef cattle, dairy cattle or sheep; class)

• Dose (g NO3/kg DM; continuous)

• Feeding management ( or ; class)

• Treatment duration (weeks; continuous)

• Treatment type (coated or plain Ca Nitrate; class)

5

Results METHB (N=21)

Overall significant increase in

MetHb% by 1.2% units

(±0.35SE; P<0.001)

Significant effect for feeding

management

Restricted fed (n=11)

• MetHb +2.4% (±0.47SE;

P<0.001)

• Max values increased at dose

>18 g NO3/kg DM

Ad libitum fed (n=10)

• MetHb +0.4% (±0.31SE;

P=0.17)

• Max MetHb level did not

exceeded 7.2%

6

0

5

10

15

20

25

30

0 5 10 15 20 25 30

Me

tHb

% M

ea

n a

nd

Ma

x

Nitrate dose (g/kg DM)

Restricted/fixed fed;

Treatment mean and maximum

0

5

10

15

20

25

30

0 5 10 15 20 25 30

Me

tHb

% M

ea

n a

nd

Ma

x


Ad libitum fed;

Treatment mean and maximum

Animal type (beef cattle, dairy

cattle or sheep)

Feeding management ( or

)

Results

METHANE (G/KG DMI; N=27)

Overall significant reduction in methane yield (-3.6 g/kg DMI; ±0.33SE;

P<0.001)

Significant effect of dose

• nitrate decreases methane by 0.21g (±0.035SE; P<0.001) per g nitrate

7

Animal type (beef cattle,

dairy cattle or sheep)

Feeding management

( or )

-9

-8

-7

-6

-5

-4

-3

-2

-1

0

0 5 10 15 20 25 30

Eff

ec

t s

ize

(T

rt -

Co

ntr

ol)


Treatment MD with nitrate dose

Results METHANE (G/KG DMI; N=27)

20.8 gram per 100 gram instead of 25.8 gram = 80.6% efficacy

No apparent decrease in methane decreasing efficacy with increasing

nitrate dose

8



Feeding management

( or )

0

0.2

0.4

0.6

0.8

1

1.2

1.4

0 5 10 15 20 25 30

eff

ica

cy (

rea

l re

du

cti

on

/sto

ich

iom

etr

ic p

ote

nti

al)

nitrate dose (g/kg DM)

Efficacy per comparison

with increasing dose of nitrate

© 2014 Cargill, Incorporated. All rights reserved.GGAA 2016-2.0

-1.5

-1.0

-0.5

0.0

0.5

1.0

1.5

2.0

0 5 10 15 20 25 30

Eff

ect

siz

e M

D (

Trt

-C

on

tro

l)

Duration of study (weeks)

Results

DMI (N=39)

Small overall negative

effect -0.12kg

(SE±0.06:P<0.003)

• No significant effect of

dose, animal type,

coating or feeding

management.

• Significant effect of exp.

Duration with decrease

in DMI with longer

experiments

9



Feeding management (

or )

-2.0

-1.5

-1.0

-0.5

0.0

0.5

1.0

1.5

2.0

0 5 10 15 20 25 30

Eff

ect

siz

e M

D (

Trt

-C

on

tro

l)


Treatment Mean Difference

Treatment MD, effect of exp. duration

Results

PH (N=11)

• Overall increase in pH with

0.06 points (SE±0.003;

P=0.003)

• Significant effect of animal

(P=0.0148) with sheep

and beef cattle showing a

significant increase, but

not in dairy cattle.

10




or )

-0.3

-0.2

-0.1

0.0

0.1

0.2

0.3

0 5 10 15 20 25 30

Ch

an

ge i

n p

H (

Trt

-C

on

tro

l)


pHTreatment MD with dose

-0.05

0.00

0.05

0.10

0.15

0.20

1

pHbeef cattle

dairy cattle

sheep

Results

TOTAL VFA

(n=11)

• Overall no effect of

nitrate on VFA

• Tendency (P=0.093) for

effect of animal type with

only sheep showing a

significant increase in

tVFA

11




or )

-50

-40

-30

-20

-10

0

10

20

30

0 5 10 15 20 25 30

Eff

ect

siz

e (

Trt

-C

on

tro

l)


tVFA

-4

-2

0

2

4

6

8

10

12

1

tVFAbeef cattle

dairy cattle

sheep

Treatment MD

Treatment MD

Remove ‘only’ before sheep

© 2014 Cargill, Incorporated. All rights reserved.GGAA 2016

Results

ACETATE %

(n=15)

• Overall nitrate increases

Acetate by 2.46%

(P<0.001)

• Effect of nitrate or coated

nitrate, with coated nitrate

not increasing Ac%

• Effect of feeding

management, with

restricted feeding

increasing Ac% more

12




or )

-2

0

2

4

6

8

10

0 5 10 15 20 25 30

Ch

an

ge i

n a

ceta

te %

(T

rt -

Co

ntr

ol)

Nitrate dose (g/kg feed DM)

Ac%Treatment

-2

-1

0

1

2

3

4

5

coated nitrate nitrate (not coated)

MD in Ac%

0

1

2

3

4

5

6

ad libitum restricted

MD in Ac%

Type of feeding managementType of nitrate

Results

PROPIONATE %

(n=15)

• Overall nitrate decreases

propionate wby 2.17% s

(P<0.001)

• Effect of type of nitrate,

with coated nitrate not

decreasing Pr%

13




or )

-12

-10

-8

-6

-4

-2

0

2

4

0 5 10 15 20 25 30

Ch

an

ge i

n p

rop

ion

ate

% (

Trt

-C

on

tro

l)


Pr%Treatment

-4

-3

-2

-1

0

1

2

coated nitrate nitrate (not coated)

MD in Pr%Type of nitrate

Results

N-BUTYRATE %

(n=15)

• Overall no effect of

nitrate on Butyrate %

(P=0.94)

• No other moderators had

an effect (e.g. dose,

duration)

14




or )

-7

-6

-5

-4

-3

-2

-1

0

1

2

3

4

0 5 10 15 20 25 30

Ch

an

ge i

n b

uty

rate

% (

Trt

-C

on

tro

l)


Bu%

Treatment

Results

AMMONIA

(n=14)

• Overall nitrate showed a

tendency to decrease

ammonia by 1.1 mmol/L

(P=0.08)

• Significant effect of

experiment duration

(P<0.001) with ammonia

decreasing with

increasing duration

15




or ) -10

-8

-6

-4

-2

0

2

4

6

0 2 4 6 8 10 12 14

ch

an

ge i

n a

mm

in

mM

(T

rt -

Co

ntr

ol)

Treatment duration (weeks)

Ammonia, mM

-8

-6

-4

-2

0

2

4

6

0 5 10 15 20 25 30

ch

an

ge i

n a

mm

in

mM

(T

rt -

Co

ntr

ol)


Ammonia, mM

Treatment effect with nitrate dose

Treatment effect with duration of experiment

ConclusionMETA-ANALYSIS

MetHb%

• When nitrate-fed ruminants are limit-fed (e.g. to 95% of ad libitum intake) mean MetHb is increased significantly by 2.4% compared to the control.

• MetHb% is not different from the control when cattle or sheep are fed ad libitum

• Risk of increased MetHb% is higher in limit fed animals

Methane Yield

• Nitrate reduced methane yield in a linear dose response manner ( 80.6% efficacy)

16

Data (appendix)INCLUDED IN THE META-ANALYSIS

18

REF-ID YEARCOMPA

-RISONCOUNTRY ANIMAL

FIXED or

AD LIBTREATMENT

DURATION

IN WEEKS

MAX*

DOSEn CH4 DMI RUMEN METHB

deRaphélisSoissan 01 2014 1 Australia Sheep Fixed Nitrate 10 20 7 x x x x

El-Zaiat 01 2014 1 Brazil Sheep Ad lib Coated nitrate 13.1 27 6 x x x x

Guyader 01 2015 1 France Dairy Fixed Nitrate 5 23 4 x x x x

Guyader 02 2015 1 France Dairy Fixed Nitrate 5 23 4 x x x x

Hulshof 01 2012 1 Brazil Beef Ad lib Nitrate 3.3 22 14 x x x

Lee 01 2015 3 Canada Beef Ad lib Coated nitrate 4 23 8 x x

Lee 02 2015 3 Canada Beef Ad lib Coated nitrate 4 23 8 x x x

Li 01 2012 1 Australia Sheep Both Nitrate 7.7 19 5 x x x x

Li 02 2013 1 Australia Sheep Both Nitrate 9.3 19 5 x x x

Newbold 01 2014 5 USA Beef Fixed Nitrate 6.1 28 6 x x x

Newbold 01 2014 5 Brazil Beef Ad lib Nitrate 15.7 24 5 x

Nolan 01 2010 1 Australia Sheep Fixed Nitrate 3 25 4 x x x x

Troy 01 2015 4 UK Beef Ad lib Nitrate 24 22 6 x x x

vanZijderveld 01 2010 1 Netherlands Sheep Fixed Nitrate 4 26 5 x x x

vanZijderveld 04 2011 1 Netherlands Dairy Fixed Nitrate 12.7 21 10 x x x

Velazco 01 2013 1 Australia Beef Ad lib Nitrate 6 19 11 x x

Veneman 01 2015 1 New Zealand Dairy Both Nitrate 4.6 20 6 x x x x

Veneman 01 2015 1 UK Dairy Ad lib Nitrate 5 20 6 x x x x

Olijhoek 01 2014 3 Denmark Dairy Ad lib Nitrate 4 21 4 x x x x

* formulated dose of nitrate in g/kg feed DM

The Effect of Dietary Nitrate on Enteric Methane Emissions ... - J. Newbold.pdf · Jolien Veneman,...

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