HEPAGEN

1. HEPAGEN Metabolic Management of Cow Health

2. MetabolicManagement of Cow Health • Cow metabolism during transition • Fatty liver and related disorders • Transcriptional control of lipid/energy metabolism: PPARs • PPAR-alpha agonists in veterinary

3. The TransitionPeriod • The last 3 wk before to 3 wk after parturition Extreme CHALLENGE Pregnant Nonlactating Nonpregnant Lactating • Tremendous metabolic adaptations to support lactation • Most diseases occur during or soon after this time

4. Hepatic Adaptation to Lactation Big changes over a very short time highlight the tremendous metabolic adaptations necessary to adequately support lactation

5. Energy intake and requirements for a lactation in dairy cows Mcal/day After parturition extra energy requirement for milk production is not met by feed energy intake Negative Positive Lipolisis Energy Balance = Energy Ingested - Energy Required DIM Parturition AdaptedfromBauman and Currie 1980

6. Lipid Metabolism during NEB Adipose Tissue ↑ epinephrine NEB HSL ↓ Insulin Lipolisis VLDL CO2 Liver CPT1 CPT2 CO2 Muscles Udder β-oxidation CoA CoA CoA CoA CoA CoA CoA KB KREB’S TG CO2 NEFA CO2

7. Blood NEFA and Liver TG around calving

8. Incidence of fatty liver in dairy cows Fatty liver is a common condition, up to 50 % of dairy cows

9. ConsequencesofFattyLiver The Liver sits at the crossroads of metabolism Its integrity is vital to all physiological processes Fatty liver has detrimental effects on health, productivity and fertility

10. Association of fatty liver with health status Bobe 2004

11. Association of Fatty Liver with impairment of the immune system Mastitis incidence (30 days) Hepatic fat increment (2 wk after vs. 2 wk before calving) Curtis 1989

12. Association of fatty liver with reproductive performance Bobe 2004

13. PPARs Regulation of Lipid Metabolism

14. MetabolismRegulation • All the cells regulate their metabolism in response to changes in the environment and metabolize fuels according to their availability • CLASSICAL VIEW • metabolic adaptations are controlled only by hormonal or neuronal signals • MODERN VIEW • Nutrients can directly regulate metabolism in a hormonal independent manner

15. Regulation of fat/cell interactions Lipids control the expression of genes involved in their own metabolism PPARs PeroxisomeProliferator-ActivatedReceptors Fatsensorstransducingchanges in cellularlipidlevelsto the transcriptionalregulationof target genesinvolved in fatty acid metabolism

16. PPARs are Nuclearreceptors • Receptors found within the nucleus • Bind directly to DNA and regulate gene expression • Ligand activated transcription factors NUCLEAR RECEPTORS

17. PPARPeroxisomeProliferator-ActivatedReceptors • Nuclearreceptorsinvolved in the transcriptionalregulationoflipidmetabolism and energybalance • Fatty acids and their derivatives (Acyl-CoA & eicosanoids) are the natural ligands of PPAR ANIMATION

18. 3 PPAR isotypesactasFatSensors Any changes in endogenous fatty acid profiles modulate the activity of PPAR PPARγ Fat Storage PPARα PPARδ Fat Catabolism

19. PPARs modulate Fat & Energy Metabolism PPARα PPARγ PPARδ FAT energy FAT energy STORAGE BURNING

20. PPAR α Acts in livertomaintain hepatic lipid homeostasis and reducesfatconcentrations Up-regulates genes involved in all aspects of Fat Catabolism NEFA Uptake Peroxisomal β-oxidation NEFA Transport Mitochondrial β-oxidation

21. BURNING LIVER FAT Adipose Tissue PPARα Activator ↑ epinephrine NEB HSL ↓ Insulin Lipolisis VLDL CO2 Liver CPT1 CPT2 CO2 Muscles Udder β-oxidation CoA CoA CoA CoA CoA CoA CoA KB KREB’S TG CO2 NEFA CO2

22. PPAR-αactivators: 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

23. CH3 O O C C OH CH3 HEPAGEN® 2-methyl-2-phenoxy-propionic acid 2-methyl-2-phenoxy-propanoic acid 2-Phenoxyisobutyric acid 2,2-Dimethylphenoxyacetic acid Mefepronic acid

24. CLINICAL TRIALS Effects of PPARα activation in dairy cows

25. EffectsofHepagen on liverfunction and fertility 40 Holsteincows (2°-5° lactation) 50 ml/cow 50 ml/cow 50 ml/cow 1 d 15 d 30 d Calving 3 d 5 d Biopsy Biopsy Biopsy • 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

26. Liver fat in control and Hepagen treated cows 1 d 15 d 30 d CONTROL TREATED P < 0.001 30 μm Liver sections stained with toluidine blue Sciorsci 2009

27. Liver glycogen in control and Hepagen treated cows 1 d 15 d 30 d CONTROL TREATED 30 μm Liver sections stained with haematoxylin-PAS to highlight the presence of glycogen (purple). Sciorsci 2009

28. Albumins – ProteinSynthesis • Albumin concentration significantly higher in the treated group • Albumin concentration in the control group slightlylowerthan the normalrange

29. Reproductive Parameters CONTROL HEPAGEN Sciorsci 2009 p<0.05

30. HepagenEffects on Ketosis 36 Holsteincows (2°- 4° lactation) 50 ml/cow 50 ml/cow 50 ml/cow Calving -6/10 d 10 d 30 d 40 d BHB BHB BHB BHB Bouda et al. 2008

31. Hepagen Effects on Ketosis p<0.05 Open days: lower in Control than in Treated group 109.9 vs. 118.5 days Bouda et al. 2008

32. HepagenEffects on Ketosis 57 PluriparousHolsteincows 50 ml/cow 50 ml/cow Calving -7/10 d -5/8 d 2 hours 2 d 10 d 21 d BHB BHB BHB BHB BHB Aparicio et al. 2009

33. Serum BHB Concentrations

34. Use Of Hepagen® in the Transition Dairy Cow : Practical Experiences 200 “Parmigiano-Reggiano” cows 50 ml/cow 50 ml/cow -20 d Calving Follow Up (postpartum diseases, fertility) BHB once/week Gorrieri 2009

35. Use Of Hepagen® in the Transition Dairy Cow : Practical Experiences metritis, displaced abomasum Gorrieri 2009

36. HEPAGEN Preventive and Therapeutic Protocols for the Transition Cow

37. Preventive protocols in close-up dry cows To complement transition cow management programs and herd preventive health care programs Reduce risk of postpartum diseases • 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

38. Preventive protocols in close-up dry cows 50 ml/cow 50 ml/cow 7-10 days before expected time of Calving Day of calving

39. Preventive Protocols in Fresh Cows Identify primary target for prevention • 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.)

40. Preventive Protocols in Fresh Cows 50 ml/cow 50 ml/cow Calving After 2-3 days

41. Therapeutic Protocols in Fresh cows Best with Fresh Cow Medicine Programs Daily Monitoring of Each Cow for First 10 Days after Calving (Temperature and Physical Exam) Early Identification and Treatment of Problem Cows

42. Diagnosis of Fatty Liver • Difficult • No specific symptoms • Diagnosed by biopsy • invasive technique • hemorrhage, infection, death • New promising ultrasound technology

43. 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

44. Therapeutic Protocols in Fresh cows 50 ml/cow 50 ml/cow Follow up and repeat where appropriate Early Identify and Treat After 24 h

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