HEPAGEN

Metabolic Management of Cow Health

Roberto Farina - Fatro

Metabolic Management of Cow Health

• Cow metabolism during transition

• Fatty liver and related disorders• Transcriptional control of lipid/energy

metabolism: PPARs

• PPAR-alpha agonists in veterinary

The Transition Period

The last 3 wk before to 3 wk after parturition

Tremendous metabolic adaptations to support lactation Most diseases occur during or soon after this time

PregnantNonlactating

NonpregnantLactating

ExtremeCHALLENGE

Hepatic Adaptation to Lactation

Prepartum Postpartum Increase

Hepatic Blood Flow 1140 l/h 2099 l/h + 84%

DMI 9.8 kg/d 14.1 kg/d + 44 %

Liver Oxygen Utilization 1619 mmol/h 3159 mmol/h + 95 %

Daily Metabolic Activity per gram of liver 4.4 mmol O2/g 8.6 mmol O2/g X 2

Glucose Release from Liver 1356 g/d 2760 g/d X 2

Big changes over a very short time highlight the tremendous metabolic adaptations necessary to adequately support lactation

Energy intake and requirements for a lactation in dairy cows

0 14 28 42 56 70 84 98112

126140

154168

182196

210224

238252

266280

294-10

0

10

20

30

40

Energy Ingested

Energy Required

Neg

ativ

e

Po

siti

ve

Energy Balance = Energy Ingested - Energy Required

DIM

Mca

l/da

y

Adapted from Bauman and Currie 1980

After parturition extra energy requirement for milk production is not met by feed energy intake

Lipolisis

Parturition

KB

AdiposeTissue

TG NEFA

VLDL

CoA

CPT1CPT2

HSL

β-oxidation

KREB’S

CoACoA

CO2

CO2

CoA

CoA

CoA

CO2

CO2Liver

MusclesUdder

CoA

↓ Insulin

↑ epinephrine

Lipolisis

NEB

Lipid Metabolism during NEB

-17 -14 -11 -8 -5 -2 1 4 7 10 13 16 19 22 250

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

0%

2%

4%

6%

8%

10%

12%

14%

NEFA Liver TG

Days

NEF

A m

Eq/L

Live

r TG

- %

DM

ParturitionAdapted from Overton 2003

Blood NEFA and Liver TG around calving

Incidence of fatty liver in dairy cows

Fatty liver is a common condition, up to 50 % of dairy cows

Consequences of Fatty Liver

Fatty liver has detrimental effects on health, productivity and fertility

The Liver sits at the crossroads of metabolism

Its integrity is vital to all physiological processes

Association of fatty liver with health status

Disorder Association ReferenceDisplaced abomasum +++ Wada et al., 1995; Rehage et al., 1996

Impaired immunoreactivity ++ Wentink et al., 1997; Zerbe et al., 2000

Ketosis +++ Gröhn et al., 1987; Veenhuizen et al., 1991

Laminitis + Fronk et al., 1980; Rehage et al., 1996

Mastitis ++ Morrow et al., 1979

Metritis ++ Haraszti et al., 1982; Heinonen et al., 1987

Milk fever + Higgins and Anderson, 1983; Gröhn et al., 1987

Retained placenta + Haraszti et al., 1982; Heinonen et al., 1987

Bobe 2004

Association of Fatty Liver with impairment of the immune system

Curtis 1989

Mas

titis

inci

denc

e (3

0 da

ys)

Hepatic fat increment(2 wk after vs. 2 wk before calving)

Association of fatty liver with reproductive performance

Parameter Association Reference

First ovarian activity ++ Reid et al., 1983; Rukkwamsuk et al., 1999c

First ovulation + Reid et al., 1983

First estrus + Paulová et al., 1990; Jorritsma et al., 2000

First insemination + Reid et al., 1983

Days open ++ Heinonen et al., 1987; Paulová et al., 1990

Pregnancy rate ++ Haraszti et al., 1982; Jorritsma et al., 2000

Services/cow + Schäfer et al., 1988; Paulová et al., 1990

Bobe 2004

PPARs

Regulation of Lipid Metabolism

Metabolism Regulation• All the cells regulate their metabolism in

response to changes in the environment and metabolize fuels according to their availability

MODERN VIEWNutrients can directly

regulate metabolism in a hormonal independent

manner

CLASSICAL VIEWmetabolic adaptations are controlled only by hormonal or neuronal

signals

Regulation of fat/cell interactions

PPARsPeroxisome Proliferator-Activated Receptors

Fat sensors transducing changes in cellular lipid levels to the transcriptional regulation of target

genes involved in fatty acid metabolism

Lipids control the expression of genes involved in their own metabolism

PPARs are Nuclear receptorsNUCLEAR RECEPTOR LIGAND

Thyroid hormone R Thyroid hormone

Glucocorticoid R Cortisol

Estrogen R Estrogen

Progesterone R Progesterone

Androgen R Testosterone

PPAR Lipids

Receptors found within the nucleus

Bind directly to DNA and regulate gene expression

Ligand activated transcription factors

NUCLEAR RECEPTORS

PPARPeroxisome Proliferator-Activated Receptors

• Nuclear receptors involved in the transcriptional regulation of lipid metabolism and energy balance

• Fatty acids and their derivatives (Acyl-CoA & eicosanoids) are the natural ligands of PPAR

ANIMATION

3 PPAR isotypes act as Fat Sensors

PPARγ

PPARδ

Fat Storage

PPARα

Fat Catabolism

Any changes in endogenous fatty acid profiles modulate the activity of PPAR

PPARs modulate Fat & Energy Metabolism

PPARα PPARδ PPARγ

STORAGE

FATenergy

BURNING

FATenergy

PPAR αActs in liver to maintain hepatic lipid

homeostasis and reduces fat concentrations

Peroxisomalβ-oxidation

Mitochondrialβ-oxidation

NEFATransport

NEFAUptake

Up-regulates genes involved in all aspects of Fat Catabolism

↑ epinephrine

↓ Insulin

KB

AdiposeTissue

TG NEFA

NEB

Lipolisis

HSL

CO2

CO2

MusclesUdder

VLDL

CoA

CPT1CPT2

β-oxidation

KREB’S

CoACoA

CO2

CO2

CoA

CoA

CoA

CoA

BURNING LIVER FAT

Liver

PPARα Activator

PPAR-α activators: Fibrates• fenofibrate, gemfibrozil• used to lower triglycerides and raise

HDL-C in dyslipidemia to reduce risk of cardiovascular events

• 2-phenoxy-2-methyl-propionic acid• Hepagen• used to treat fatty liver, related metabolic

disorders and improve energy balance

HEPAGEN®

2-methyl-2-phenoxy-propionic acid

2-methyl-2-phenoxy-propanoic acid2-Phenoxyisobutyric acid2,2-Dimethylphenoxyacetic acidMefepronic acid

O C

CH3

CH3

C

OH

O

CLINICAL TRIALSEffects of PPARα

activation in dairy cows

Effects of Hepagen on liver function and fertility

40 Holstein cows (2°-5° lactation)

• Treated group: 50 ml of Hepagen® I.M. at calving, 3d postpartum and 5d postpartum• Control group: 50 ml of physiological solution (NaCl 0.9%)/ head at calving, 3d

postpartum and 5d postpartum

50 m

l/co

w

50 m

l/co

w

50 m

l/co

w

Calv

ing 3 d 5 d

Biopsy

1 d

Biopsy

Biopsy

15 d 30 d

Liver fat in control and Hepagen treated cows

Liver sections stained with toluidine blue

Sciorsci 2009

30 μm

CO

NT

RO

LT

RE

AT

ED

1 d 15 d 30 d

15 d 30 d0%

1%

2%

3%

4%

5%

6%

7%

8%

9%

10%

8.6%9.0%

0.3% 0.1%

Liver Fat

ControlTreated

P < 0.001

Liver glycogen in control and Hepagen treated cows

Liver sections stained with haematoxylin-PAS to highlight the presence of glycogen (purple). Sciorsci 2009

30 μm

CO

NT

RO

LT

RE

AT

ED

1 d 15 d 30 d

Albumins – Protein Synthesis

1 3 5 10 15 30 4026

27

28

29

30

31

32

33

34

35

36

ControlLinear (Control)HepagenLinear (Hepagen)

Days from parturition

g/L

• Albumin concentration significantly higher in the treated group• Albumin concentration in the control group slightly lower than the normal range

Reproductive Parameters

11 d 13 d -

1 2 3 4 5 6 7

3.70 4.24 5,46

6.47

Progesterone

ng/m

l

-

20

40

60

80

74 50

Days to 1° Heat

days

- 20 40 60 80

100 120 140

12192

Days open

days

-

20

40

60

80

50 6457 71

Pregnancy rate

days

Sciorsci 2009p<0.05HEPAGENCONTROL

Hepagen Effects on Ketosis36 Holstein cows (2°- 4° lactation)

50 m

l/co

w

50 m

l/co

w

50 m

l/co

w

Calv

ing

10 d-6/10 d

BHB

30 d 40 d

BHB

BHB

BHB

Bouda et al. 2008

Hepagen Effects on Ketosis

-10 10 30 400.40.50.60.70.80.91.01.11.2

Serum BHB Concentrations

BHB

mm

ol/L

10 30 400%

25%

50%

44%39%

28%17%

6% 6%

Subclinical Ketosis BHB > 1.4

CONTROLHEPAGEN

Bouda et al. 2008

p<0.05

Open days: lower in Control than in Treated group 109.9 vs. 118.5 days

Hepagen Effects on Ketosis57 Pluriparous Holstein cows

50 m

l/co

w

50 m

l/co

w

Calv

ing

2 hours-7/10 d

BHB

10 d 21 d

BHB

BHB

Aparicio et al. 2009

-5/8 d

BHB

2 dBH

B

CONTROL BCS: 3.25 – 3.75 TREATED BCS: 3.25 – 3.75

CONTROL BCS: ≥ 4 TREATED BCS: ≥ 4

Serum BHB Concentrations

-10 -7 2 10 210

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

CONTROL BCS ≥ 4

CONTROL BCS: 3.25 – 3.75

HEPAGEN BCS ≥ 4

HEPAGEN BCS: 3.25 – 3.75

Use Of Hepagen® in the Transition Dairy Cow : Practical Experiences

200 “Parmigiano-Reggiano” cows

50 m

l/co

w

50 m

l/co

wCa

lvin

g-20 d

Gorrieri 2009

BHB once/week

Follow Up (postpartum diseases, fertility)

Use Of Hepagen® in the Transition Dairy Cow : Practical Experiences

0%5%

10%15%20%25%30%35%

30 %15 %

Postpartum Diseases

0.00.20.40.60.81.0

0.86

0.35

Milk BHB

BHB

mg/

dl

50 60 70 80 90

100

8976

Days open

days

0%5%

10%15%20%25%30%

25 %

10 %

BHB +

metritis, displaced abomasum

Gorrieri 2009

HEPAGENPreventive and

Therapeutic Protocols for the Transition Cow

Preventive protocols in close-up dry cows

Monitor and record for diseases occurring during the early lactation period in the herd:• Lactational incidence risk (LIR): #affected / # of calvings (at

risk) in the same time period•Case definitions/Confidence of diagnosis grade•Define targets for acceptable levels of incidence

Reduce risk of postpartum diseases

To complement transition cow management programs and herd preventive health care programs

50

ml/

cow

7-10 days before expected time of Calving

50

ml/

cow

Day of calving

Preventive protocols in close-up dry cows

Preventive Protocols in Fresh Cows

Cows at a higher risk of fatty liver and metabolic disordes: Over-conditioned Underfed Quick weight loss Calving difficulties, Twins Predisposing diseases (Infections, RP, etc.)

Identify primary target for prevention

50

ml/

cow

Calving

50

ml/

cow

After 2-3 days

Preventive Protocols in Fresh Cows

Therapeutic Protocols in Fresh cows

Daily Monitoring of Each Cow for First 10 Days after Calving

(Temperature and Physical Exam)

Early Identification and Treatment of Problem Cows

Best with Fresh Cow Medicine Programs

Diagnosis of Fatty Liver

• Difficult • No specific symptoms• Diagnosed by biopsy – invasive technique–hemorrhage, infection, death

• New promising ultrasound technology

Diagnosis of Fatty Liver• Cows having problems from the beginning of

lactation• Rapid weight and BCS loss, reduced feed intake• Presence of ther diseases• Diseases more severe and less responsive– Milk fever cows that relapse and become downers– Ketotic cows that don’t respond to treatment– Chronic mastitis cows– Repeat breeders that defy all treatments– Cows that relapse or go from one disease to another– Reduced milk production– Cows that are frequently culled

50

ml/

cow

Early Identify and Treat

Follow up and repeat where appropriate

50

ml/

cow

After 24 h

Therapeutic Protocols in Fresh cows

THANK YOU