research on herbal drugs for poultry & livestock
DESCRIPTION
RESEARCH ON HERBAL DRUGS FOR POULTRY & LIVESTOCK. COLLEGE OF VETERINARY MEDICINE. BACKGROUND INFORMATION. COLLEGE OF VETERINARY MEDICINE. Two researches on local herbal plants conducted. Herbal drugs for Haemophilus paragallinarum infection. - PowerPoint PPT PresentationTRANSCRIPT
RESEARCH ON HERBAL DRUGS FOR POULTRY & LIVESTOCK
BACKGROUND INFORMATION
• Two researches on local herbal plants conducted
Herbal drugs for Haemophilus paragallinarum infection
Herbal drugs for Haemonchus contortus infection
OBJECTIVES OF TWO RESEARCHES
• 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
• Determine the shelf-life of plant drugs
• Find out the quality of binders of tablet forms of the plant drugs for infectious coryza
MATERIALS AND METHODS
Steps:
• Consultation with farmers
• Identification of local plants
• Diagnosis of diseases
• Culture of causative agents of diseases
• In vitro assay of local plants for their bactericidal/larvicidal actions
• Pharmacologic studies of herbal drugs
• Quality control study of herbal drugs
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
Identification of local plants used by farmers
Preparation & dosage level of a particular herb
Diagnosis of diseases based on clinical signs
• Isolation of Haemonchus contortus
Blood agar/nutrient broth culture
• Isolation of Haemophilus paragallinarum in pure culture
Gram staining & Biochemical tests
Culture of eggs & infection of goats
Infection and isolation of H. paragallinarum
Culture of Infective Stage for In Vitro Assay
Blood agar
• Stock of pure culture of H. paragallinarum
• Supply of eggs for culture of H. contortus
Infection of goats with L3s for supply of eggs
Identification of Infected Goats
by fecalysis
Collection of Fecal Sample
Fecalysis (Floatation Technique)
Infected GoatsIdentified
Culture Feces for L3sof H. contortus
Retrieval & Identification of L3s
L3s of H. contortus
3 Goats InfectedWith L3s*
*Source of eggs for culture of L3s
Collect feces
isolation of eggs of H. contortus
Culture of Haemonchus larvae
Moistened cotton wads placed at the bottom of shot glass
Feces from infected goats macerated and placed on top of cotton wads
Sides of shot glass filled with tap water
Culture of Haemonchus larvae
Shot glass in Culture glass, filled with water up to the brim of shot glass
Culture glass covered with wrappings andincubated at room temperature for 6 days
Recovery of Infected Larvae (L3) of H. Contortus
Removal of wrappings of shot glass
Water in the culture glass transferred into petri plate
Infective Larvae (L3s) recovered in Petri plate identified
Preparation of Plant Extracts:
• Decoction using 1:2 (w/v)
• Dried plant parts were boiled for 15 minutes
• Crude extracts were filtered
• 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.
IN VITRO ASSAYS OF LOCAL PLANT EXTRACTS
Techniques Used in In Vitro Assay
• Sensitivity test of Carter for Haemophilus gallinarum
• Larvicidal test for Haemophilus gallinarum
RESULTS
Plants and Parts used by local farmers for the treatment against H. paragallinarum infection in chicken (Fernandez, 1990)
Scientific Names Common Names
Local Names Plant Parts
Ananas squamosa Sweet Sop Atis Leaves
Capsicum frutescens Wild Pepper Siling labuyo Fruits
Chrysanthemum indicum
Mansaninya Hilbas Leaves
Citrus grandis Pomelo Buongon Rinds
Coleus aromaticus Oregano Karabo Leaves
Coleus blumei - Mayana (red) Leaves
Helianthus annuus* Sunflower Sunflower Seeds
Heliotropium indicum Elephant Grass
Trompa Elefante
Leaves
Spondias pinnata - Libas Leaves
Solanum spp. - Terramycin Plant
Fruits
Zingiber officinale Ginger Luya Root
*Fruits given raw to sick birds
In vitro assay of plant extracts* vs H. paragallinarum
**Sensi discs, BBL Microbiological systems
Treatment No.
Plant ExtractsZone of Inhibition (cm)
After 1 h
After 24 h
T0(-) Distilled Water NZ NZ
TO(+) Streptomycin disc** 3 5
T1 Anona squamosa 1 NZ
T2 Capsicum frutescens NZ NZ
T3 Chrysanthemum indicum
NZ NZ
T4 Citrus grandis 1.5 NZ
T5 Coleus aromaticus NZ NZ
T6 Coleus blumei NZ NZ
T7 Helianthus annuus NZ NZ
T8 Heliotropium indicum 3 6
T9 Spondias pinnata 2 3
T10 Solanum spp. NZ NZ
T11 Zingiber officinale 1 NZ*1:2 (w/v) 1 part plant part: 2 parts distilled water by decoction, amount in each wellwas 0.05ml
In vivo assay of plant extracts* against H. paragallinarum infection in chicken (Lohman)
*1:2 (w/v) 1 part plant part: 2 parts distilled water by decoction, amount given to each bird was 5ml, given per os
Treatment No.
Plants No. of Birds
Examined
Water Eye & Nasal
Discharge
Facial Swelling
Mucus in Upper Respiratory
Tract
Congested Lungs
TO Untreated Control 4 4 4 4 4
T1 Anona squamosa 6 4 3 3 0
T2 Capsicum frutescens 6 3 6 6 3
T3 Chrysanthemum indicum
6 4 5 5 2
T4 Citrus grandis 6 3 3 3 0
T5 Coleus aromaticus 5 4 4 4 0
T6 Coleus blumei 6 2 2 2 1
T7 Helianthus annuus 5 2 2 2 1
T8 Heliotropium indicum 5 0 0 0 0
T9 Spondias pinnata 6 3 3 3 0
T10 Solanum spp. 6 3 4 4 0
T11 Zingiber officinale 5 1 2 2 0
Chicken Showing Clinical Signs of Haemophilus paragallinarum Infection
Chicken Showing Clinical Signs of Haemophilus paragallinarum Infection
Percent efficacy* of plant water extract by decoction according to animalspecies against Haemonchus contortus (Fernandez, 1991)
PlantsPlantsChicken Chicken
(% (% Efficacy)Efficacy)
Swine (% Swine (% Efficacy)Efficacy)
Goat (% Goat (% Efficacy)Efficacy)
Ananas comosus (fruit)Ananas comosus (fruit) 63.363.3 59.159.1 52.352.3
Ananas comosus Ananas comosus (leaves)(leaves)
NINI 54..754..7 56.756.7
Mangifera indicaMangifera indica 60.060.0 58.258.2 56.756.7
Manihot esculentaManihot esculenta 53.453.4 52.052.0 50.850.8
Tamarindus indicaTamarindus indica 59.459.4 58.458.4 55.655.6
Moringa oleiferaMoringa oleifera 69.069.0 64.764.7 61.561.5
Artemisia vulgarisArtemisia vulgaris 57.657.6 56.356.3 53.353.3
Mimosa pudicaMimosa pudica NINI NINI 79.7**79.7**
Chrysophyllum cainitoChrysophyllum cainito NINI NINI 70.3**70.3**
Tinospora rumphiiTinospora rumphii NINI NINI 85.6***85.6***
NI = not used as anthelmintic by local farmers in a particular animal species
*Based on formula of Reik and Keitz (1954)
Group
No.*
Scientific Names (LocalNames)
Plant Parts Preparati
ons
Dosages Administration
Chicken
Swine Goat
TO Control** - - - - - -
T1 Ananas comosus (pina)
Unripe fruit
pounding 15 g 30 g 40g Mixed w/ feed
T2 Artemisia vulgaris (hilbas)
leaves decoction
15 ml 30 ml 40 ml Per os
T3 Bixa orellana (Asuite)
seeds decoction
5 ml 10 ml 20 ml Per os
T4 Cajanus cajan(Kadios)
roots decoction
15 ml 30 ml 40 ml Per os
T5 Cassia alata(sunting)
seeds pounding 15 g 30 g 40 g Per os
T6 Chrysophylum cainito (caimito)
leaves decoction
- - 40 ml Per os
T7 Clitorea ternatea (balog-balog)
seeds heating &
pounding
15 g 30 g 40 g Mixed w/ feed
Preparations, dosage levels and mode of administration of plant parts usedby local farmers against parasitic infections in various animals
*No. of Animals/treatment = 4
**Untreated control
- not used by farmers
Group
No.*
Scientific Names (LocalNames)
Plant Parts Preparati
ons
Dosages Administration
Chicken
Swine Goat
T8 Karicq papaya(kapayas)
seeds pounding 15 g - 40 g Per os
T9 Lansium domesticum (lansones)
seeds pounding 15 g 30 g 40 g Per os
T10 Leucaena leucocephala
(ipil-ipil)
seeds Pounding 15 g 30 g 40 g Per os
T11 Mangifera indica (paho)
seeds Pounding 15 g 10 g 30 g Per os
T12 Manihot esculenta
(balanghoy)
bark/ roots
Decoction 15 ml 30 ml 40 ml Per os
T13 Mimosa pudica(makahiya)
leaves decoction - - 40 ml Per os
T14 Momordica charantia (paliya)
leaves Heating/ expressin
g
3 ml 6 ml 12 ml Per os
T15 Moringa oleifera (malunggay)
seeds pounding 15 g 30 g 40 g Per os
Preparations, dosage levels and mode of administration of plant parts usedby local farmers against parasitic infections in various animals
*No. of Animals/treatment = 4
- not used by farmers
Preparations, dosage levels and mode of administration of plant parts usedby local farmers against parasitic infections in various animals
*No. of Animals/treatment = 4
- not used by farmers
Group No.*
Scientific Name (Local
Name)
Plant Parts
PreparationDosages
Administration
chicken Swine
Goat
T16 Quisqualis indica(niyog-
niyogan)
seeds pounding 15 g 30 g 40 g Per os
T17 Tamarindus indica
(Sambag)
leaves decoction 15 ml 30 ml 40 ml Per os
T18 Tinospora rumphii
(panyawan)
stem decoction - - 40 ml Per os
Percent efficacy of plant parts according to animal species
Plants Chicken (% Efficacy)
Swine (% Efficacy)
Goat (% Efficacy)
Control -7.1 -13.7 -7.8
Ananas comosus 63.3 59.1 52.3
Artememisia vulgaris 57.6 56.3 53.3
Bixa orellana 66.0 61.4 58.3
Cajanus cajan 57.7 53.3 53.3
Cassia alata 62.8 59.1 57.0
Chrysophyllum cainito NI NI 70.3
Clitorea ternatea 62.8 58.7 55.6
Karica papaya 59.4 NI 58.8
NI = not used by local farmers
*highly effective
Percent efficacy of plant parts according to animal species
Plants Chicken (% Efficacy)
Swine (% Efficacy)
Goat (% Efficacy)
Lansium domesticuml 65.6 61.0 60.7
Leucaena leucocephala
68.0 62.1 62.1
Mangifera indica 60.0 58.2 56.7
Manihot esculenta 53.4 52.0 50.8
Mimosa pudica NI NI 79.7
Momordica charantia 74.7 70.0 64.5
Moringa oleifera 69.0 64.7 61.5
Clitorea ternatea 62.8 58.7 55.6
Quisqualis indica 70.7 70.0 62.4
NI = not used by local farmers
Percent efficacy of plant parts according to animal species
NI = not used by local farmers
*highly effective
Plants Chicken (% Efficacy)
Swine (% Efficacy)
Goat (% Efficacy)
Tamarindus indica
59.4 58.4 55.6
Tinospora rumphii
NI NI 85.6*
On-Going Study on Haemonchus Contortus
Rationale
Goat Population:
As of 2002, 3.29 million heads
25% are concentrated in the Visayas
99.9% of 3.29 millions are raised by small holders ( in rural areas)
Source: Philippines Recommends for Goat Production, PCARRD (2004)
Constraints in Goats Production:
Endoparasitism hampers productivity & full development of goat subsector
Ways endoparasites hampered productivity of goats:
2. Drop in milk production
1. High rate mortality at weaning
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)
Haemonchus contortus is one of these endoparasites considered most pathogenic (Soulsby, 1982)
Two classes of anthelmintic that could be resorted to:
Haemonchus contortus infection in ruminants is addressed by use of athelmintic or dewormers
1. Commercial or synthetic anthelmintic
2. Herbal anthelmintic
Advantage of synthetic anthelmintic:
1. Being pure, synthetic anthelmintic is very effective
5. Not available in remote areas
Advantage is outweighed by the following:
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
Use of alternative dewormer: Herbal anthelmintic
Advantages of Herbal anthelmintic:
1. Does not impart drug residue in meat
2. Not very expensive
3. Available all year round
4. Parasite does not develop resistance
5. No chemical that would pollute environment
Requisites for an Effective Dewormer:
1. Kills all worm burden (Cytotoxic action)
2. Expels dead worm (Cathartic or purgative)
3. Heals injury brought about by inflammatory reaction by worms (Astringent)
Compounds in 3 Plants with Their Corresponding Compounds and actions:
PlantsPlantsCompounds & ConcentrationCompounds & Concentration
FlavonoiFlavonoidsds
AnthraquinoAnthraquinonesnes
AlkaloiAlkaloidsds
TanniTanninsns
Chrysophyllum Chrysophyllum cainitocainito
++++ -- ++++++++ --
Tinospora Tinospora rhumpiirhumpii
++++++++ -- ++++ --
Mimosa Mimosa pudicapudica
-- ++++ -- ++++++++
C. caimitoM. pudicaT. rumphii
(Ratio of Extracts)
FlavonoidsTannins
AnthraquinonesAlkaloids
Model Animal
Monoclonal Antibodies
Isolate
Bioactive Compounds
Antigen
Compounds
Inject Hyperimmunize
Hybridoma Cell /
E. coli
Anthelmintic activity
Purified dewormer
Testing
Final product
Hatch
L1 and L2
Develop
L3 on Grass
Ingested
Goat Infected
Feces
Developmental Cycle of Haemonchus spp.
Developmental Cycle of Haemonchus contortus
Eggs in Pasture area
L1 and L2
L1 and L2
Purposes of Phase I
• The effective concentration against L3s of H. contortus will be the basis for the determination of LD50 of plant cocktail
• Extraction by solvent will be the basis for isolation of bioactive compounds
• Effective plant cocktail dewormer will be made into drug forms and these will be studied for their ED50
L1 and L2
Objectives of Phase I
• 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
General:
• Determine the ratios of the plant cocktail that would kill at least 80% of the L3s of H. contortus
Specific:
• 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
MATERIALS AND METHODS
Collection of Plant Parts
Air-Drying of T. rumphii StemChopped Stem of T. rumphii
Air-Drying of C. cainito Leaves Chopped leaves of C. cainito
Leaves of Mimosa pudica Leaves of Chrysophyllum cainito
Stem of Tinospora rumphii
Plant Sample
Petroleum Ether
Petroleum Ether Extract
Crude PetroleumEther Fraction
Assay*
Rotavap
Plant Residue
Ethanol
Ethanol Extract
Crude Ethanol Fraction
Assay*
Rotavap
Plant Residue
Water
Water Extract
Crude Aqueous Fraction
Assay*
Rotavap
Plant Residue
Discard
*Highly effective extract by solvent will be combined for herbal cocktail
Extraction of Individual Plant Parts
Combination of Plant Extracts*
Assay of Plant Cocktail forlarvicidal activity
Effective Combination of Plant Cocktail against L3
Dosage Level for LD50 and ED50
Studies (Phase II)
Isolation of Bioactive Compounds (Phase II)
*Highly Effective Extract by Solvent will Be combined to form a plant cocktail
Multiplication of Bioactive Compounds
Weighing of Plant Parts
Transfer of Plant Parts in Amber Bottles
Extraction by Infusion
Straining of Plant Part Extracts
Setting up of Rotavapor
Concentration of Plant Part Extracts in Rotavapor
Plant Extracts Ready for Concentration in a Rotary evaporator
Plant Extracts by Solvents & 0.5% Ivermectin
Tinospora rumphiiTinospora rumphii Chrysophyllum cainitoChrysophyllum cainito Mimosa pudicaMimosa pudica
1 part1 part 1 part1 part 1 part1 part
1 part1 part 1 part1 part 2 parts2 parts
1 part1 part 1 part1 part 3 parts3 parts
1 part1 part 2 parts2 parts 1 part1 part
1 part1 part 2 parts2 parts 2 parts2 parts
1 part1 part 2 parts2 parts 3 parts3 parts
1 part1 part 3 parts3 parts 1 part1 part
1 part1 part 3 parts3 parts 2 parts2 parts
1 part1 part 3 parts3 parts 3 parts3 parts
2 parts2 parts 1 part1 part 1 part1 part
2 parts2 parts 1 part1 part 2parts2parts
Table 1. Varying ratios used in combining the 3 plants (makabuhay, caimito, and makahiya) extracts
Table 1. Continued
2 parts2 parts 1 part1 part 3 parts3 parts
2 parts2 parts 2 parts2 parts 1 part1 part
2 parts2 parts 2 parts2 parts 3 parts3 parts
2 parts2 parts 3 parts3 parts 1 part1 part
2 parts2 parts 3 parts3 parts 2 parts2 parts
2 part2 part 3 parts3 parts 3 parts3 parts
3 parts3 parts 1 part1 part 1 part1 part
3 parts3 parts 1 part1 part 2 parts2 parts
3 parts3 parts 1 part1 part 3 parts3 parts
3 parts3 parts 2 part2 part 1 part1 part
3 parts3 parts 2 parts2 parts 2 parts2 parts
3 parts3 parts 2 parts2 parts 3 parts3 parts
3 parts3 parts 3 parts3 parts 1 part1 part
3 parts3 parts 3 parts3 parts 2 parts2 parts
A S S A Y OF PLANT EXTRACTS
L3s Transferred into Wells of Hollow Glass Slide
Exposure of L3s with Plant Extracts
L3s in wells of hollow glass slide counted
L3s in wells of hollow glass slide exposed with0.5 ml of the varying concentrations of plantcocktail
Movement of L3s monitored for 30 Minutes after exposure
Absence of movement upon touchingof L3s with end of inoculating needle
Exposure of L3s with Plant Extracts
Computation for the Dosage Level of the Combined Plant Extracts:
A x 100 B
Where:
A = Weight (mg/g) of Combined Plant Extract
B = Volume (ml) of Distilled water
% Efficacy = No. of Dead L3s x 100 No. of L3s Exposed
Where: Below 70 % efficacy, the plant extract is said to be ineffective;
71%-80 % efficacy, the plant extract is said to be effective
81-100% efficacy, the plant extract is said to be highly effective
Formula of Reik and Keitz (1954) on Percent Efficacy of Anthelmintic
Experimental Design:
Experimental Design:
Layout of Experiment on Plant Cocktails: Completely Randomized Design (CRD)
TO(-) = 1% Ethanol
TO(+) = 0.5% ivermectin
Treated Groups =nth concentrations of plant cocktail
Replicates = 2 with at least 30 L3s per replicate
Layout of experiment of Individual Plant Extracts: Completely Randomized Design (CRD)
T0(-) = 1% of appropriate solvent
T0(+) = 0.5% Ivermectin
Treated Groups: nth Concentrations of appropriate extract
Replicates: 2 with at least 30 L3s per replicate
Statistical Analysis:
Experiment on Plant Cocktails:
Experiment of Individual Plant Extracts:
Analysis of Variance (ANOVA)
Significant among treatment means: DMRT
Analysis of Variance (ANOVA)
R E S U L T S
Mean Percent Efficacy of Individual Panyawan-, Caimito-, & Makahiya- Pet Ether Extracts
0102030405060708090
1000.
5%Iv
erm
ectin
1% P
etEt
her
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Concentrations
% E
ffica
cy
PANYAWAN
CAIMITO
MAKAHIYA
Mean Percent Efficacy of Individual Panyawan-, Caimito-, & Makahiya- Ethanol- Extracts
0
10
20
30
40
50
60
70
80
90
100
0.5%
Iver
mec
tin1%
Pet
Ethe
r
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Concentrations
% E
ffica
cy
PANYAWAN
CAIMITO
MAKAHIYA
Mean Percent Efficacy of Individual Panyawan-, Caimito-, & Makahiya- Water- Extracts
0102030405060708090
1000.
5%Iv
erm
ectin
1% P
etEt
her
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Concentrations
% E
ffica
cy
PANYAWAN
CAIMITO
MAKAHIYA
Petroleum Ether-Ethanol Extraction
Treatments Treatments (P:C:M)(P:C:M)
MeanMean
TT0(+) 0(+) (0.5% (0.5%
Ivermectin)Ivermectin)
37.90b37.90b
TT0(-)0(-) (1% Ethanol)(1% Ethanol) 14.83c14.83c
TT1 1 (1:1:1)(1:1:1) 97.88a97.88a
TT2 2 (1:1:2)(1:1:2) 100.00a100.00a
TT3 3 (1:1:3)(1:1:3) 100.00a100.00a
TT4 4 (1:2:1)(1:2:1) 100.00a100.00a
TT5 5 (1:2:2)(1:2:2) 100.00a100.00a
TT6 6 (1:2:3)(1:2:3) 100.00a100.00a
TT7 7 (1:3:1)(1:3:1) 100.00a100.00a
TT8 8 (1:3:2)(1:3:2) 100.00a100.00a
TT9 9 (1:3:3)(1:3:3) 100.00a100.00a
TT10 10 (2:1:1)(2:1:1) 100.00a100.00a
TT11 11 (2:1:2)(2:1:2) 100.00a100.00a
TreatmentsTreatments
MeanMean
TT12 12 (2:1:3)(2:1:3) 100.00a100.00a
TT13 13 (2:2:1)(2:2:1) 98.04a98.04a
TT1414 (2:2:3)(2:2:3) 98.60a98.60a
TT15 15 (2:3:1)(2:3:1) 100.00a100.00a
TT16 16 (2:3:2)(2:3:2) 98.96a98.96a
TT17 17 (2:3:3)(2:3:3) 98.15a98.15a
TT18 18 (3:1:1)(3:1:1) 100.00a100.00a
TT1919 (3:1:2)(3:1:2) 99.02a99.02a
TT20 20 (3:1:3)(3:1:3) 98.99a98.99a
TT21 21 (3:2:1)(3:2:1) 100.00a100.00a
TT22 22 (3:2:2)(3:2:2) 99.12a99.12a
TT23 23 (3:2:3)(3:2:3) 100.00a100.00a
TT24 24 (3:3:1)(3:3:1) 100.00a100.00a
TT25 25 (3:3:2)(3:3:2) 100.00a100.00a
Means with different letters are statistically significant (p<0.01) by DMRT
Plant Cocktail by Petroleum Ether-Ethanol Extraction
Mean Percent Efficacy of Pet Ether to Ethanol- Plant- Cocktail Extracts at 100% Concentration
05
101520253035404550556065707580859095
100
0.5%
Iver
mec
tin
1% E
than
ol
T1 T2 T3 T4 T5 T6 T7 T8 T9
T10
T11
T12
T13
T14
T15
T16
T17
T18
T19
T20
T21
T22
T23
T24
T25
Treatments
% E
ffica
cy
Mean Percent Efficacy of Ethanol- Plant Cocktail Extracts at 100% Concentration
05
101520253035404550556065707580859095
100
0.5%
Iver
mec
tin
1% E
than
ol T1 T2 T3 T4 T5 T6 T7 T8 T9
T10
T11
T12
T13
T14
T15
T16
T17
T18
T19
T20
T21
T22
T23
T24
T25
Treatments
% E
ffica
cyPlant Cocktail by Ethanol Extraction
PHASE II ACTIVITIES
Pharmacologic Studies of Plant Cocktail
• LD50 of plant cocktail
• ED50 of plant cocktail
Studies on Drug Forms
• Studies on the appropriate binders for tablet forms of plant cocktail
• Studies on the appropriate capsules for plant cocktail
Pharmacologic Studies of Plant Cocktail
• LD50 of plant cocktail
• ED50 of plant cocktail
Studies on Drug Forms
• Studies on the appropriate binders for tablet forms of plant cocktail
• Studies on the appropriate capsules for plant cocktail
Bioactive Compounds Elucidation
Plant Extracts
Isolate
FlavonoidsTannins
AnthraquinoneAlkaloids
Inject
Animal Model
MonoclonalAntibodies
Bioactive Compounds.
Isolate
Bioactive Compounds Elucidation
Solid Phase
Ligand
Antibody
Antigen
Bioactive Compounds Elucidation
Antigen (bioactive compounds)
Hybridoma/E. coli
Purified Anthelmintic
Final Product
Inoculate
Testing
Testing
Plantdrug
dev’t.
HybridomalE. coli
Monoclonalantibodies
Bioactivecmpds
Postmarket
Mktgsales
Pilotcomm’lprod’n
Qualitycontrol
Field evl’n.
Herbal Anthelmintic Product Path
IP/Biosafety Audit
Biosafety Approval
Regulatory approvals(BFAD)
DiscoveryProduct
Development Product Enhancement
Yr 1 Yr 1 - 3 Yr 2 Yr 3 Yr 4
Pure Form
Plant/Raw
MaterialsScreening
Labevl’n.
Crude Form
Drug Forms
Drug Forms