diagnosis and (early) prediction of acidosis and ketosis using ...acidosis and ketosis using milk...

1

Veerle Fievez

Ghent University

Laboratory for Animal Nutrition and Animal Product Quality

Diagnosis and (early) prediction of

acidosis and ketosis using milk fatty

acids as biomarkers

Overview

• Fatty acids

• Origin of milk fatty acids

• Milk fatty acids as biomarkers

• Negative energy balance & ketosis

• Acidosis

• Potential and constraints for practice


http://www.lanupro.UGent.be – [email protected]

2

1

2 3 4

a b g

Essential fatty acids

Fatty acids

Overview

• Fatty acids




• Acidosis




3



Milk fat composition



Milk fatty acids

4

Overview

• Fatty acids




• Acidosis




Overview

• Fatty acids



• Negative energy balance & ketosis - Metabolism of mobilized fat

- Composition of mobilized fat

- Potential as biomarker

• Acidosis




5

Laboratory for Animal Nutrition and Animal Product

Quality

http://www.lanupro.UGent.be –

[email protected]

Metabolism of

mobilised fat



Composition of

mobilised fat

Subcutaneous

Visceral

Hostens et al.2012. J Dairy Sci 95,

3756.

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Potential as biomarker

of NEB – why ?

-Metritis

-(Sub)clinical ketosis

-Displaced abomasum -Retained placenta

risk

Economic losses

-Increased culling

-Reduced fertility

-Less milk

Metabolism of

mobilised fat



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C18:1cis-9: potential

biomarker of NEB

β2 1+exp(- β0*(x- β1))

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NEF

A

Milk fat C18:1 cis-9 g/100g milk fatty acids

β1= 23.5

FP = 6.5% (Sp = 93.5%)

Blue colour = ill cow

Green colour = healthy cow

Jorjong et al.2014. J Dairy Sci 97, 7054.



C18:1cis-9: potential

early biomarker of NEB

0.0

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Pro

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NEF

A

Milk fat C18:1 cis-9 (g/100 g milk fatty acids)

β1= 24

FP = 11.4% (Sp = 88.6%)

Blue colour = ill cow

Green colour = healthy cow

Jorjong et al.2014. J Dairy Sci 97, 7054.

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Validation C18:1cis-9:

biomarker of NEB Accuracy of the prediction Sensitivity (Se), Specificity (Sp)

biomarker

High plasma NEFA??

Diagnosis

MF C18:1 c-9 of 23




biomarker of NEB

MF C18:1 c-9 NEFA≥0,6 NEFA<0,6

≥24

<24

Jorjong et al. In preparation.

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biomarker of NEB

Se = TP/(TP+FN)

Sp =

TN/(TN+FP)

MF C18:1 c-9 NEFA≥0,6 NEFA<0,6

≥24

TP

FP

<24

FN

TN

Jorjong et al. In preparation.




biomarker of NEB Se =

TP/(TP+FN)

Sp =

TN/(TN+FP)

MF C18:1cis-9

< or ≥ 4.0

≥ 4.1

Experiment 1 (d14) 55 89

Experiment 2 (d14) 58 87

Experiment 3 (d10)

(d3)

45

81

84

75

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Laboratory for Animal Nutrition and Animal Product

Quality

http://www.lanupro.UGent.be –

[email protected]

Metabolism of

mobilised fat



Biomarker of

propionate supply

linear odd-chain (C15:0, C17:0)

iso branched-chain (even & odd)

anteiso branched-chain (odd)

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Bacterial OBCFA &

rumen end products




http://www.lanupro.UGent.be – [email protected] 21



A, P/S, L

Cellulolytic

Amylolytic

Sugar

Pectin

Fievez et al.2012. Anim Feed Sci

Technol 172, 51.







C18:1cis-9/C15:0:

potential biomarker SCK FP = 17% (Sp = 83%)

FN = 20% (Se= 80%)

Jorjong et al. J Dairy Sci, revised

version submitted

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Biomarker SCK –

available cow-side tests

Product Detection level of

BHBA

Sensitivity (%) Specificity (%)

KetoCheck Powder

(milk or urine)

Trace 41 99

KetoStix (urine) Small 78 96

KetoTest (milk) 0.2 mmol/L 27-59 76-99

PortaBHB (milk) 0.2 mmol/L 75 91

Precision Xtra (blood) 1.4 mmol/L 91 94

Towsend et al. 2011

Overview

• Fatty acids




• Acidosis




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Overview

• Fatty acids




• Subacute rumen acidosis (SARA) - Rumen metabolism of fatty acids

- Fatty acids associated with low rumen pH

- Potential as biomarker






Overview – rumen

metabolism

1

14

Bacterial OBCFA &

rumen end products







A, P/S, L

Cellulolytic

Amylolytic

Sugar

Pectin

Fievez et al.2012. Anim Feed Sci

Technol 172, 51.



Overview – rumen

metabolism

1

2 3

4

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Lipolysis

Lipolysis = prerequisite for BH = detoxification process

C18:0C18:0

C18:1C18:1t11t11

C18:2C18:2t11c15t11c15

C18:3C18:3c9t11c15c9t11c15

C18:3C18:3c9c12c15c9c12c15

C18:0C18:0

C18:1C18:1t11t11

C18:2C18:2t11c15t11c15

C18:3C18:3c9t11c15c9t11c15

C18:3C18:3c9c12c15c9c12c15

Rumen BH:

C18:2n-6 : 87 ± 7 %

C18:3n-3 : 86 ± 7 %

C20:5n-3 : > 90 %

C22:6n-3: > 90 %

Glasser et al. 2008. Animal 2, 691.

Chilliard et al. 2010. OCL 17, 22.

Biohydrogenation



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Rumen

biohydrogenation

Chilliard et al. 2007. Eur. J. Lipid Sci. Technol.

109, 828.

SLOW

FAST



Rumen biohydro-

genation: low pH

Chilliard et al. 2007. Eur. J. Lipid Sci. Technol.

109, 828.

Partial inhibition trans 18:1

reductase

Secundary pathway



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Biomarker of SARA



Colman et al. Comp Electron Agric, in press.

acidosis classification models based on pH-criterion – OBCFA & specific trans-FA

(pH < 5.6 for at least 3h/d; pH < 5.6 for at least 4.5h/d; pH < 5.8 for at least 8h/d)

iso C14:0

iso C15:0

iso C16:0

iso C17:0

anteiso C13:0

anteiso C15:0

anteiso C17:0

C15:0

C17:0

C18:1trans-10

C18:2cis-9,trans-11

C18:1trans-11

FA in same column do not appear simultaneously in model

FA presented in white – positively related to acidosis risk

FA presented in yellow – negatively related to acidosis risk

Validation milk FA

biomarker of SARA



Colman et al. Comp Electron Agric, in press

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Overview

• Fatty acids




• Acidosis




Prerequisites for

transfer to practice



Routine analysis

Improvement models (Se & Sp)

Monitoring & feedback

http://www.google.be/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&cad=rja&uact=8&ved=0CAcQjRw&url=http%3A%2F%2Foffline070.com%2F1566%2Fgo-de-a13-krijgt-een-online-community&ei=Nll8VKe8FcLkaKu3gOAK&psig=AFQjCNErz8tm7MQSA9Fglawj1-hR0fj7pQ&ust=1417521746751467

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Routine analysis:

raw milk



0.00

0.10

0.20

0.30

0.40

0.50

0.60

0.70

0.80

0.90

1.00

Rcv2

NEB

Hyperketonemia

SARA

Stefanov, 2012. PhD dissertation UGent.

Routine analysis:

milk fat trans



NEB

Hyperketonemia

(SARA)

0.00

0.10

0.20

0.30

0.40

0.50

0.60

0.70

0.80

0.90

1.00

Rcv2

Stefanov et al. 2013. J Agric Food Chem, 61,

3403.

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Routine analysis:

milk fat OBCFA



NEB

Hyperketonemia

SARA

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0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

Rcv

2

Stefanov et al. 2010. J Agric Food Chem, 58,

10804.

Monitoring:

economics - NEB



• Incidence – prevalence

• Treatment cost

• ‘Disease’ cost

• Model performance (Se & Sp)

‘Cheap to treat’ ‘Disease is not

associated with

major loss’

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Monitoring:

economics



De Ridder, 2014. MSc dissertation UGent.

20-24 Euro/case

Conclusion



Milk fatty acids - biomarkers of metabolic disorders

1/ NEB !

2/ Hyperketonemia: C15:0 = challenge

3/ SARA: routine analysis = challenge (OBCFA)

4/ Economics: semi-routine & specific groups @ risk

diagnosis and (early) prediction of acidosis and ketosis using ...acidosis and ketosis using milk...

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