Clinical Testing and Therapeutic Application of Formulated Bee

Department of Agriculture
BUREAU OF AGRICULTURAL RESEARCH
Apitherapy in Animals:
Clinical Testing and Therapeutic
Application of Formulated Bee Products
as an Antimicrobial and Wound Healing
Enhancing Agent Using Animal Models
8 December 2014
Clark, Pampanga
1
PROPONENTS: Maria Amelita C. Estacio, DVM, PhD
Waren N. Baticados, DVM, PhD
Therese Marie A. Collantes, DVM, MS
Jussiaea V. Bariuan, DVM
Mark Joseph M. Desamero, DVM
Abigail Baticados, DVM
TARGET PROJECT COST: 750,000.00 Php
TOTAL AMOUNT SPENT: 750,139.12 Php
TOTAL AMOUNT RELEASED: 750,000.00 Php
BALANCE: -139.12 Php
DURATION NUMBER OF YEARS: 18 months
DATE STARTED: August 1, 2012
DATE ENDED: February 28, 2014
2
PROJECT OVERVIEW
General
Objectives:
1. To compare the efficacy of ethanol extracted
propolis with the conventional antibiotic dressing in
incisional
wound
healing.
2. To compare the efficacy of newly developed
gamma-radiated propolis-alginate patch with the
antibiotic-laden patch in incisional and sutured
wounds.
3. To compare the efficacy of newly developed
gamma-radiated honey-alginate patch with the
antibiotic-laden patch in incisional and sutured
wounds.
3
PROJECT OVERVIEW
Specific
Objectives:
1. To determine the rate of incisional wound healing
among the controls and 30% ethanol extracted
propolis
(30%
EEBP)
treated
group.
2. To determine the rate of incisional & sutured
wound healing among the controls and honey-, and
propolis
wound
patch
treated
groups.
3. To characterize the quality of post-operative
wound resolution among controls and 30% EEBP
treated
group
by
gross
examination.
4. To characterize the quality of incisional and
sutured wound resolution among the controls and
honey, and propolis wound patches treated groups by
gross
examination.
4
Specific
Objectives:
5. To characterize the histologic changes in
incisional wound among the controls and 30%
ethanol extracted bee propolis (30% EEBP) treated
group.
6. To characterize the histologic changes in
incisional and sutured wounds among the controls
& honey-, & propolis wound patch treated groups.
7. To determine the antimicrobial activity of the
30% EEBP compared with the controls by
evaluating the minimum inhibitory concentration
and antibiotic sensitivity testing.
5
PHYSICAL and FINANCIAL ACCOMPLISHMENTS
List of Activities
Accomplished
Proposed
Budget
Actual
Release
Difference
1. Antimicrobial
testing of 30%
ethanol extracted
bee propolis (30%
EEBP)
100%
440,974.00
441,113.03
-139.03
2. Efficacy testing of
30% EEBP for
incisional wound
100%
3. Efficacy testing of
honey & propolis
wound patch for
incisional & sutured
wounds: Gross and
histopathologic
evaluation
100%
TOTAL:
100%
440,974.00
441,113.03
-139.03
6
HIGHLIGHTS OF ACCOMPLISHMENTS
Project Component
and Activities per
Approved WP
Activities Undertaken
Approved
Budget
Actual
Expenses
Variance
1. Established the
minimum effective
concentration of
propolis for wound
management
2. Established the
antimicrobial and wound
healing activities of 30%
ethanol extracted bee
propolis (30% EEBP)
a. Bacterial isolation & identification
from wound & adjacent skin of test cats
b. Conducted MIC of 30% EEBP using
lab vs. wound & skin bacterial isolates
c. Screening & acclimation of test cats
d. Induction of incisional wound in cats
& treatment with 30% EEBP
e. Gross & microscopic evaluation of
post-operative wound-treated 30%
EEBP
440,974.00
441,113.03
-139.03
3. Development of
wound patches and
evaluation of healing
properties of gammaradiated honey- &
propolis-alginate wound
patches
a. Purchasing & acclimation of mice
b. Anesthetization & induction of
incisional & sutured wounds in mice
c. Treatment of incisional & sutured
wounds with propolis & honey patches
d. Gross & microscopic evaluation of
incisional & sutured wounds treated
with honey & propolis patches
440,974.00
441,113.03
-139.03
TOTAL:
7
PHYSICAL TARGETS
- Determination
of
minimum
inhibitory
concentration of propolis extract
- Determination of antibiotic sensitivity testing of
propolis extract
- Selection of honey and propolis extract for wound
patch development
- Screening & acclimation of test cats for wound
healing efficacy testing
- Procurement & acclimation of laboratory mice for
wound healing efficacy testing
- Wound healing efficacy test in cats
- Wound healing efficacy tests in laboratory mice
- Histopathological processing and microscopic
examination of wound healing sites
8
Title ID: _______ Proponent: _________ Cost: ______ Duration: _________
MFO, Components, Activities
Physical Target and
Accomp.
Total
Targets
First Second
Year
Year
22Financial Allocation
(M PhP)
Total
Budget
First
Year
Second
Year
MFO
Major Activities
1. Antimicrobial analysis of
propolis extract
1
1
- wound & skin swabs
- bacterial isolation & ID
- MIC
- Antibiotic sensitivity tests
2. Clinical Application
- propolis extract in wounds in
cats
- honey-alginate wound
patch in wounds lab mice
- propolis-alginate wound
patch in wounds in lab mice
3. Presentation in scientific
conference
4. Published abstracts
440,974.00 441,113.03
3
1
2
3
1
2
0.00
0.00
0.00
6
0.00
0.00
0.00
6
9
PROJECT MILESTONES
Project Milestones
Brief Description
1. Establishment of minimum
effective concentration of
propolis extract with
antimicrobial and wound
healing activities
Selected propolis from the Philippine stingless bee
(Tetragona biroi) possessed microbial inhibitory
effect against the common pus forming bacteria of
contaminated wounds. In vivo testing in cats also
show increased healing rate of incisional wound.
2. Development & testing of
gamma-radiated propolisalginate patch for wound
management in humans and
animals
In vivo testing of propolis-alginate wound patch in small
lab animals (UPCVM) and preliminary testing in humans
(PNRI, East Avenue Hospital, NKI, Heart Center)
3. Development & testing of
In vivo testing of honey-alginate wound patch in small
gamma-radiated honey-alginate lab animals (UPCVM) and preliminary testing in humans
patch for for wound
(PNRI, East Avenue Hospital, NKI, Heart Center)
management in humans and
animals
4. Development of new
biomedical product for
humans and animals
Potential usage in the Philippines and abroad for
management of different types of wound
10
COMMERCIALIZATION ACTIVITIES
Standardization of medical grade honey and propolis
with PNRI
- Discussion on establishment of harvesting and
extraction protocols for medical grade honey and
propolis for development of new biomedical products
following FDA and International standards.
Wound Patch Product Development with PNRI
- Improvements on the physical properties of honey- &
propolis-alginate patches
- Improvement of the prototype wound patches with
probable inclusion of adhesives and/or sensors to
determine state of wound healing
- Improvement of storage packaging material for the
wound patches to prevent any untoward effects of sunlight
and prolonging the shelf-life.
11
COMMERCIALIZATION ACTIVITIES
Scouting Partner Veterinary & Human
Pharmaceutical Companies
- Discussion on the potential of these
wound patches with interested veterinary
and human pharmaceutical companies
12
PROJECT IMPACT
- Creation of new biomedical products
at par with international standards
- Commercialization opportunities
(for domestic and international market)
- Conservation strategies and
sustainable apiculture for the Philippine
stingless bee
- Livelihood opportunities for bee
13
PROJECT IMPACT
List of Beneficiaries
Highlights of Success Stories
1. human patients
2. medical doctors
3. military personnel
4. animal patients
5. veterinarians
6. human and veterinary
pharmaceutical companies
7. bee keepers
8. Researchers, students
2 graduated DVM thesis students
6 published abstracts
14
PROJECT SUSTAINABILITY
Sustainable
- Increasing demand for new biomedical products
(wound patches) due to increasing antibiotic
resistance
- Higher compliance and faster healing rate of
wounds for human patients especially indigents
under normal circumstances & during calamities
- High market value of new biomedical products
- Potential usage in human wound management
- Potential usage in animal wound management
- Potential interest of veterinary and human
pharmaceutical companies
15
ISSUES/CONCERNS/CHALLENGES &
ACTIONS TAKEN
Issues / Concerns/
Challenges
Action Undertaken
1. Availability & staggered
procurement of test mice
Two months advance
reservation of mice to the
officer-in-charge of the
laboratory animal facility
16
PHOTOS (with captions)
Please include the following:
1.
2.
3.
4.
5.
Project site/s and signage
Products/By-products
Action shots of project team, farmers, processors
Snap shot of the Audited Financial Report
PDF file of the complete list of beneficiaries
17
Apitherapy Project Sites
Laboratory Animal Experimental Room & Microbiology
Laboratory, College of Veterinary Medicine, UPLB
18
Apitherapy Project Signage
Apitherapy project signage at the Laboratory Animal
Experimental Room, College of Veterinary Medicine, UPLB.
19
Apitherapy Project Sites
Laboratory Animal Experimental Room, College of Veterinary
Medicine, UPLB for in vivo wound healing experimentation
20
Antimicrobial Activity of 30%
Ethanol Extracted Bee Propolis
21
22
23
24
25
Wound Healing Activity of 30% Ethanol Extracted
Bee Propolis (30% EEBP)
26
Developed & Tested Gamma Radiated Honey& Propolis-Alginate Wound Patches
27
Representative Mouse per Treatment Group with
Complete Incisional Wound Healing
28
Representative Mouse per Treatment Group with
Incomplete Incisional Wound Healing
29
30
H2O2 + Povidone iodine
H2O2 +
Povidone iodine
Alginate
Fusidate sodium
Honey-alginate
Propolis-alginate
Normal skin
of Mouse
31
32
Representative Mouse per Treatment Group with
Complete Sutured Wound Healing
33
Representative Mouse per Treatment Group with
Incomplete Sutured Wound Healing
34
35
H2O2 +
Povidone iodine
Fusidate
sodium
Propolisalginate
Alginate
Honeyalginate
Normal skin
of Mouse
36
37
Audited Financial Report
38
END
39

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