SELAMAT

DATANG

Frog Skin and Its

Potential in Wound Dr Ahmad Hata Rasit
Professor of orthopedics

Healing Faculty of Medicine & Health Sciences

Universiti Malaysia Sarawak

www.unimas.my
INTRODUCTION
WHY Odorrana hosii (Boulenger,
1891)?
MOST HANDSOME FROG

POISON FROG
Odorrana hosii
Dorsal top is dark green with
both lateral sides marked with
brown lines while the ventral
side is silvery and yellowish
white. Its hind legs were
stripes of light and dark brown.
-skin is potential for
wound healingQ

-microhabitat
specialist so prone to
be threatened by
habitat conversion
 Research Questions

01 Water Quality
Is the distribution of O hosii skin
glands affected by water quality?

02 Frog Skin Histology

Is skin histology different
depending on skin region and
sampling location?

03 Wound Healing
Do wounds on the skin
regions heal at different
rates?
Research Objectives
Water Quality
To compare water quality
parameters in various
locations and their effects on
skin glands.

Frog Skin Histology

To compare the histology of
frog skin at various skin regions
and localities.

Wound Healing
To compare wound healing in
the skin regions on Days 3, 7,
and 14.
 Sampling Site (Localities)
The frogs, O. hosii, were collected in
four locations: (1) Ranchan
Recreational Park, (2) Matang Wildlife Santubong National Park Matang Wildlife Center
Center, (3) Borneo Highland, and (4) (1.7° N, 110.3325° E) (1.6° N, 110.1591° E)
Santubong National Park, with
permission from the Sarawak
Biodiversity Council and Sarawak
Forestry Department. Borneo Highland
(1.5° N 110.15° E) Ranchan Recreational Park
(1.15° N, 110.583° E)

Water samples were collected and tested on-site

using two pieces of equipment: a pH metre (Eutech
pH 150) and a waterproof handheld turbidimeter
(Eutech ECTN 100 IR).
Methods
The use of Wistar rats in
live experiments was
approved by the Universiti
Malaysia Sarawak Animal

H
Frog Skin histology Ethics Committee
based on skin (UNIMAS-AEC) and was
regions overseen by authorities
throughout the

G
Gland distribution experiment.
based on skin
regions

E Excision
wound model

H
Histology on
wound
healing
 The mucous glands are identified with flatten
nuclei at the apical of the single simple
squamous or cuboidal cells
Mucosal glands secrete mucus to maintain the
moisture, permeability and elasticity of the
skin, all of which are necessary for amphibian
homeostasis 

The seromucous glands, also known as

mixed glands, exhibit the properties of
both mucous glands and granular glands

Granular glands (poison glands) are

characterized by the flatten nuclei in the cells
which line the acinus

Figure : 40× magnification of O. hosii skin showing

The chromatophore are pigment cells found
detailed cutaneous glands. M – Mucous gland; SM –
under the basal of the epidermal cells which
Seromucous gland; G – Granular gland and Chr –
gives the colouration of the frog
Chromatophore (pigment cells).
Histology of wound healing

Figure : 40× magnification of rat’s wound. (1) Wound

length; (2) Dermal length; (3) Epidermis length; (4)
Epidermis; (5) Dermis; (6) Hypodermis; (7) Granulation
tissue; and (8) Scab.
Wound Healing Parameter ( Gal et al., 2008)

• Epidermal closure (quantitative) Qualitative scale for inflammation rate modified from Gal et al. (2008).

Scale Epithelization PMNL Fibroblasts New Vessels Hair Follicle

 100%
0 thickness of cut edge absent absent absent absent

• Dermal closure (quantitative) 1 migration of cells (<50%) mild ST mild ST mild SCT <30%

2 migration of cells (>50%) mild DL/GT mild GT mild GT >50%

• Granulation tissues (quantitative)
3 bridging the excision moderate DL/GT moderate GT moderate GT >80%

 100% 4 keratinization marked DL/GT marked GT marked GT mature follicle

Semi-quantitative evaluation of histological sections (ST - surrounding tissues; DL -

• Inflammation rate (semi quantitative) demarcation line; SCT - subcutaneous tissue; GT -granulation tissue)
RESULTS
1.Water Quality Parameters
30.0000 p=0.00*

25.0000

20.0000

p=0.087 p=0.00*
15.0000

10.0000 p=0.00*

5.0000

0.0000
pH Turbidity Temperature Dissolved Oxygen
The distribution of mucous and seromucous glands appears to be
influenced by dissolved oxygen (DO), but not that of granular
glands.

1. RRP has the lowest DO followed by MWC, BH and SNP. A trend is noted where O. hosii that dwells in low
DO waters have high number of seromucous glands. O. hosii found in RRP and BH requires more
adaptation mechanism for a higher AMP and homeostasis where waters are low in DO. A low DO may
indicate stagnant or still waters where a high amount of microbial activities may take place. Seromucous
glands serve to function as O. hosii’s defense against the external environment and to facilitate oxygen
diffusion into the skin.

2. Number of mucous glands was found to be the highest in O. hosii from BH and SNP where DO is high. The
frogs adapt to needing less of secretion of AMP from seromucous glands that exhibits both mucous glands
and granular glands properties. Hence, having high number of mucous glands is enough to ensure its
survivability in the environment.
 NOTES TAKEN
1. Mucous and seromous glands are essential for preventing skin dehydration in frogs. Mucosal
glands secrete mucus to maintain the moisture, permeability and elasticity of the skin,
all of which are necessary for amphibian homeostasis
Habit and Habitat of O. hosii (Inger et al 2017)
2. Dissolved oxygen (DO) is the most important indicator of freshwater quality because it is
required
1. Neverfor amphibians
found more thanandfewother
metresaquatic life to grow
from edge/rocky (Wahaba et al, 2019; Fitri et al.
streams
2021).
2. MoreVariations
common inin primary
DO concentration can have
forest, also lives an impact
in partially loggedon freshwater
forest providedbiological
a swift and
chemical
clear processes (Fitri et al. 2021). High DO is found in clear and fast flowing streams
stream is available
(O.
3. hosii
Perchmicrohabitats), which trees
on rocks /shrubs/small contain the moststreams,
overhanging oxygen.particularly where the current
is strongest
3. 4. Skin
Water secretions
quality can kill small
parameters haveanimals, such
no effect onasthe
other frogs, not of
distribution dangerous
glanulartoglands
human (important
though.healing)
for wound But if webut
were
thetohigh
be absent-mindedly
distribution ofrub
theour eyes in
glands after
O. handling O hosii,
hosii makes thethe
species
result is quite painful. Wash hands thoroughly after handling frogs
suitable for wound healing management.

4. As a result, we must protect O. hosii's microhabitats for its survival so that we can explore
more on the species' full potential as a wound healer.
2.Histology comparisons from skin regions,
study locations, and frog skin layers
120.000 p=0.00* p=0.16* p=0.00*

100.000

80.000

60.000

40.000

20.000

0.000
Ranchan Matang Borneo Santubong Ranchan Matang Borneo Santubong Ranchan Matang Borneo Santubong
Highlands Highlands Highlands
Epidermis Spongiosum Compactum

Dorsal Head Dorsal Bottom Dorsal Thigh Ventral Head Ventral Bottom Ventral Thigh

Thickness of skin between four localities and six skin regions (µm).
 2.Histology comparisons from skin regions,
study locations, and frog skin layers
4.00 p=0.014*
p=0.333 p=0.000*

3.50

3.00

2.50

2.00

1.50

1.00

.50

.00
Ranchan Matang Borneo Santubong Ranchan Matang Borneo Santubong Ranchan Matang Borneo Santubong
Highlands Highlands Highlands
Mucous Seromucous Granular

Number of glands between four localities and six skin regions.

 2.Histology comparisons from skin regions,
study locations, and frog skin layers
25000
p=0.005* p=0.076 p=0.059 p=0.000*

20000

15000

10000

5000

0
Borneo Highlands

Borneo Highlands

Borneo Highlands

Borneo Highlands
Matang

Santubong

Matang

Santubong

Matang

Santubong

Matang

Santubong
Ranchan

Ranchan

Ranchan

Ranchan
Mucous Seromucous Granular Chromatophore

Average size of glands between four localities and six skin regions (µm²).
3.Excisional wound healing comparison
3.Surface wound closure on Day 3 between
frog skin region and localities (%).
3.Surface wound closure on Day 7
between frog skin region and localities (%).
3.Surface wound closure on Day 14 between
frog skin region and localities (%).
3.Day 3 of epidermal closure between
frog skin region and localities (%).
3.Day 7 of epidermal closure between
frog skin region and localities (%).
3.Day 14 of epidermal closure between
frog skin region and localities (%).
3.Day 3 of dermal closure between frog
skin region and localities (%).
3.Day 7 of dermal closure between frog
skin region and localities (%).
3.Day 14 of dermal closure between
frog skin region and localities (%).
3.Day 3 granulation tissue area (%) between
frog skin region and localities.
3.Day 7 granulation tissue area (%)
between frog skin region and localities.
3. Day 14 granulation tissue area (%)
between frog skin region and localities.
 Conclusion
The dorsal region has
The study identified O. hosii skin from four the most granular
two different water different locations was glands and the fewest
quality classes, with discovered to have mucous glands,
Ranchan Recreational cutaneous gland whereas the ventral
Park being the most variations that reflect region has the most
polluted (Class IV) of its environment in seromucous glands
the four study sites terms of numeration, and fewest mucous
(Class I) size, and distribution. glands.

F W A
 Further Recommendations

Instead of suturing It is possible to

investigate the Bioprospecting of
frog skin grafts onto
wounds, frog lipid direct application of biotechnological
skin extract could be purified wound
applied topically. antimicrobial management
peptides.

L P W
Thank You

Email
rahata@unimas.
my

Phone
+60128581469