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RESEARCH ON HERBAL DRUGS FOR POULTRY & LIVESTOCK

Explore two research studies conducted at the College of Veterinary Medicine on herbal drugs for Haemophilus paragallinarum and Haemonchus contortus infections in poultry and livestock. Objectives, methods, and results are detailed.

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RESEARCH ON HERBAL DRUGS FOR POULTRY & LIVESTOCK

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  1. RESEARCH ON HERBAL DRUGS FOR POULTRY & LIVESTOCK COLLEGE OF VETERINARY MEDICINE

  2. BACKGROUND INFORMATION COLLEGE OF VETERINARY MEDICINE

  3. Two researches on local herbal plants conducted • Herbal drugs for Haemophilus paragallinarum infection • Herbal drugs for Haemonchus contortus infection COLLEGE OF VETERINARY MEDICINE

  4. OBJECTIVES OF TWO RESEARCHES COLLEGE OF VETERINARY MEDICINE

  5. Screen the inhibitory activity of local plants used by local farmers • against infectious coryza/haemonchusis • Find out the local practices adopted by farmers in using plants • for infectious coryza/haemonchusis • Determine the LD50 and ED50 of plant drugs COLLEGE OF VETERINARY MEDICINE • Find out the quality of binders of tablet forms • of the plant drugs for infectious coryza • Determine the shelf-life of plant drugs

  6. MATERIALS AND METHODS COLLEGE OF VETERINARY MEDICINE

  7. Steps: • Consultation with farmers • Identification of local plants • Diagnosis of diseases • Culture of causative agents of diseases COLLEGE OF VETERINARY MEDICINE • In vitro assay of local plants for their bactericidal/larvicidal actions • Pharmacologic studies of herbal drugs • Quality control study of herbal drugs

  8. Farmer Consultations • Interview with local farmers on: • Clinical signs of a particular disease in poultry/livestock • Local plants for treatment of a particular disease in poultry/livestock COLLEGE OF VETERINARY MEDICINE • Preparation & dosage level of a particular herb Identification of local plants used by farmers

  9. Diagnosis of diseases based on clinical signs • Isolation of Haemophilus paragallinarum in pure culture • Blood agar/nutrient broth culture • Gram staining & Biochemical tests COLLEGE OF VETERINARY MEDICINE • Infection and isolation of H. paragallinarum • Isolation of Haemonchus contortus • Culture of eggs & infection of goats

  10. Culture of Infective Stage for In Vitro Assay • Stock of pure culture of H. paragallinarum • Blood agar • Supply of eggs for culture of H. contortus COLLEGE OF VETERINARY MEDICINE • Infection of goats with L3s for supply of eggs

  11. isolation of eggs of H. contortus Identification of Infected Goats by fecalysis Collection of Fecal Sample Fecalysis (Floatation Technique) Infected Goats Identified Collect feces Culture Feces for L3s of H. contortus COLLEGE OF VETERINARY MEDICINE Retrieval & Identification of L3s L3s of H. contortus 3 Goats Infected With L3s* *Source of eggs for culture of L3s

  12. COLLEGE OF VETERINARY MEDICINE

  13. Culture of Haemonchus larvae Moistened cotton wads placed at the bottom of shot glass COLLEGE OF VETERINARY MEDICINE Feces from infected goats macerated and placed on top of cotton wads Sides of shot glass filled with tap water

  14. Culture of Haemonchus larvae Shot glass in Culture glass, filled with water up to the brim of shot glass COLLEGE OF VETERINARY MEDICINE Culture glass covered with wrappings and incubated at room temperature for 6 days

  15. Recovery of Infected Larvae (L3) of H. Contortus Removal of wrappings of shot glass COLLEGE OF VETERINARY MEDICINE Water in the culture glass transferred into petri plate Infective Larvae (L3s) recovered in Petri plate identified

  16. COLLEGE OF VETERINARY MEDICINE

  17. Preparation of Plant Extracts: • Decoction using 1:2 (w/v) • Dried plant parts were boiled for 15 minutes • Crude extracts were filtered COLLEGE OF VETERINARY MEDICINE • Filtrate was concentrated in a rotavapor (40oC) until • volume was reduced to 1/3 of its original volume • Concentrated plant extract was placed in the oven (60oC) overnight • Plant residues were collected & kept in the fridge for 1 wk.

  18. IN VITRO ASSAYS OF LOCAL PLANT EXTRACTS COLLEGE OF VETERINARY MEDICINE

  19. Techniques Used in In Vitro Assay • Sensitivity test of Carter for Haemophilus gallinarum • Larvicidal test for Haemophilus gallinarum COLLEGE OF VETERINARY MEDICINE

  20. RESULTS COLLEGE OF VETERINARY MEDICINE

  21. Plants and Parts used by local farmers for the treatment against H. paragallinarum infection in chicken (Fernandez, 1990) COLLEGE OF VETERINARY MEDICINE *Fruits given raw to sick birds

  22. In vitro assay of plant extracts* vs H. paragallinarum COLLEGE OF VETERINARY MEDICINE *1:2 (w/v) 1 part plant part: 2 parts distilled water by decoction, amount in each wellwas 0.05ml **Sensi discs, BBL Microbiological systems

  23. In vivo assay of plant extracts* against H. paragallinarum infection in chicken (Lohman) COLLEGE OF VETERINARY MEDICINE *1:2 (w/v) 1 part plant part: 2 parts distilled water by decoction, amount given to each bird was 5ml, given per os

  24. COLLEGE OF VETERINARY MEDICINE

  25. COLLEGE OF VETERINARY MEDICINE Chicken Showing Clinical Signs of Haemophilus paragallinarum Infection

  26. COLLEGE OF VETERINARY MEDICINE Chicken Showing Clinical Signs of Haemophilus paragallinarum Infection

  27. Percent efficacy* of plant water extract by decoction according to animal species against Haemonchus contortus (Fernandez, 1991) COLLEGE OF VETERINARY MEDICINE *Based on formula of Reik and Keitz (1954) NI = not used as anthelmintic by local farmers in a particular animal species

  28. Preparations, dosage levels and mode of administration of plant parts used by local farmers against parasitic infections in various animals COLLEGE OF VETERINARY MEDICINE *No. of Animals/treatment = 4 **Untreated control - not used by farmers

  29. Preparations, dosage levels and mode of administration of plant parts used by local farmers against parasitic infections in various animals COLLEGE OF VETERINARY MEDICINE *No. of Animals/treatment = 4 - not used by farmers

  30. Preparations, dosage levels and mode of administration of plant parts used by local farmers against parasitic infections in various animals COLLEGE OF VETERINARY MEDICINE *No. of Animals/treatment = 4 - not used by farmers

  31. Percent efficacy of plant parts according to animal species COLLEGE OF VETERINARY MEDICINE NI = not used by local farmers *highly effective

  32. Percent efficacy of plant parts according to animal species COLLEGE OF VETERINARY MEDICINE NI = not used by local farmers

  33. Percent efficacy of plant parts according to animal species COLLEGE OF VETERINARY MEDICINE NI = not used by local farmers *highly effective

  34. On-Going Study on Haemonchus Contortus COLLEGE OF VETERINARY MEDICINE

  35. Rationale COLLEGE OF VETERINARY MEDICINE

  36. Goat Population: As of 2002, 3.29 million heads 25% are concentrated in the Visayas COLLEGE OF VETERINARY MEDICINE 99.9% of 3.29 millions are raised by small holders ( in rural areas) Source: Philippines Recommends for Goat Production, PCARRD (2004)

  37. Constraints in Goats Production: Endoparasitism hampers productivity & full development of goat subsector Ways endoparasites hampered productivity of goats: 1. High rate mortality at weaning COLLEGE OF VETERINARY MEDICINE 2. Drop in milk production 3. Reduced feed conversion efficiency 4. Mortality of productive goats Economic Impact due to roundworm infection in goats is valued at US$3.55 M annually Source: Philippines Recommends for Goat Production, PCARRD (2004)

  38. Haemonchus contortus is one of these endoparasites considered most pathogenic (Soulsby, 1982) Haemonchus contortus infection in ruminants is addressed by use of athelmintic or dewormers COLLEGE OF VETERINARY MEDICINE Two classes of anthelmintic that could be resorted to: 1. Commercial or synthetic anthelmintic 2. Herbal anthelmintic

  39. Advantage of synthetic anthelmintic: 1. Being pure, synthetic anthelmintic is very effective Advantage is outweighed by the following: COLLEGE OF VETERINARY MEDICINE 1. Imparts residue to meat which is hazardous to consuming public 2. Parasite develops resistance against synthetic anthelmintic 3. Synthetic anthelmintic pollutes environment 4. Expensive for small holders, being imported 5. Not available in remote areas

  40. Use of alternative dewormer: Herbal anthelmintic Advantages of Herbal anthelmintic: • Does not impart drug residue in meat 2. Not very expensive COLLEGE OF VETERINARY MEDICINE 3. Available all year round 4. Parasite does not develop resistance 5. No chemical that would pollute environment

  41. Requisites for an Effective Dewormer: • Kills all worm burden (Cytotoxic action) 2. Expels dead worm (Cathartic or purgative) 3. Heals injury brought about by inflammatory reaction by worms (Astringent) COLLEGE OF VETERINARY MEDICINE Compounds in 3 Plants with Their Corresponding Compounds and actions:

  42. C. caimito M. pudica T. rumphii (Ratio of Extracts) Flavonoids Tannins Anthraquinones Alkaloids Model Animal Monoclonal Antibodies Inject Hyperimmunize Isolate Compounds Bioactive Compounds Antigen Hybridoma Cell / E. coli Testing Anthelmintic activity Final product Purified dewormer

  43. Feces Eggs in Pasture area Goat Infected Hatch COLLEGE OF VETERINARY MEDICINE Ingested L1 and L2 L1 and L2 L3 on Grass Develop Developmental Cycle of Haemonchus spp. Developmental Cycle of Haemonchus contortus

  44. Purposes of Phase I • The effective concentration against L3s of H. contortus will • be the basis for the determination of LD50 of plant cocktail • Effective plant cocktail dewormer will be made into drug forms and • these will be studied for their ED50 COLLEGE OF VETERINARY MEDICINE • Extraction by solvent will be the basis for isolation of bioactive • compounds L1 and L2

  45. Objectives of Phase I General: • Evaluate the effect of different solvents on extraction of active • compounds of the plant extract that would kill at least 80% of • L3s of H. contortus • Determine the ratios of the plant cocktail that would kill at least • 80% of the L3s of H. contortus COLLEGE OF VETERINARY MEDICINE Specific: L1 and L2 • Find out the concentration of the individual plant according to • solvent that would kill at least 80% of L3s of H. contortus • Determine the concentration of the plant cocktail that would kill • at least 80% of L3s of H. contortus

  46. MATERIALS AND METHODS COLLEGE OF VETERINARY MEDICINE

  47. Collection of Plant Parts COLLEGE OF VETERINARY MEDICINE

  48. COLLEGE OF VETERINARY MEDICINE Air-Drying of T. rumphii Stem Chopped Stem of T. rumphii Chopped leaves of C. cainito Air-Drying of C. cainito Leaves

  49. COLLEGE OF VETERINARY MEDICINE

  50. Leaves of Chrysophyllum cainito Leaves of Mimosa pudica COLLEGE OF VETERINARY MEDICINE Stem of Tinospora rumphii

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