Emu Oil

Its Anti-Inflammatory
Properties

A report for the Rural Industries Research

and Development Corporation
by J M Snowden, P J OMalley and T M Ellis
Agriculture Western Australia

October 1999

RIRDC Publication No 99/133

RIRDC Project No DAW-82A

1999 Rural Industries Research and Development Corporation.

All rights reserved.
ISBN 0 642 57954 7
ISSN 1440-6845
Emu Oil Its Anit-Inflammatory Properties
Publication no 99/133
Project no. DAW-82A.
The views expressed and the conclusions reached in this publication are those of the author and not
necessarily those of persons consulted. RIRDC shall not be responsible in any way whatsoever to any person
who relies in whole or in part on the contents of this report.
This publication is copyright. However, RIRDC encourages wide dissemination of its research, providing the
Corporation is clearly acknowledged. For any other enquiries concerning reproduction, contact the
Publications Manager on phone 02 6272 3186.

Researcher Contact Details

Dr John M. Snowden
Agriculture Western Australia
Locked Bag 4
Bentley Delivery Centre
WA 6983
Phone: (08) 9368 3555
Fax:
(08) 9474 2479
Email:
[email protected]
:

RIRDC Contact Details

Rural Industries Research and Development Corporation
Level 1, AMA House
42 Macquarie Street
BARTON ACT 2600
PO Box 4776
KINGSTON ACT 2604
Phone:
Fax:
Email:
Website:

02 6272 4539
02 6272 5877
[email protected]
http://www.rirdc.gov.au

Published in October 1999

Printed on environmentally friendly paper by Canprint

ii

FOREWORD
High prices were paid for emu fat because emu oil was believed to have therapeutic
activity. In Australia and many overseas countries it is an offence to make any therapeutic
claims for a product until its efficacy has been demonstrated and the product registered by
the relevant authority. Therefore to expand the therapeutic use of emu oil, registration as an
active ingredient under the Therapeutic Goods Act is required. The first step towards
registration is to demonstrate the efficacy of emu oil in a scientifically acceptable manner.
This report complements other RIRDC research projects investigating the potential
therapeutic applications of emu oil. These are the anti-inflammatory properties of emu oil,
the effects of emu oil on wound healing and cellular regeneration and the anti-viral/antibacterial activity of emu oil.
This publication describes an investigation into the anti-inflammatory properties of
topically applied emu oil. The other reports are also available as RIRDC research
publications.
The report, a new addition to RIRDCs diverse range of almost 400 research publications,
is part of our New Animal Industries Program which aims to accelerate the development of
viable new animal product industries.
Most of our publications are available for viewing, downloading or purchasing online
through our website:
downloads at www.rirdc.gov.au/reports/Index.htm
purchases at www.rirdc.gov.au/pub/cat/contents.html
Peter Core
Managing Director
Rural Industries Research and Development Corporation

iii

Contents
Foreword

iii

Executive Summary

Introduction

Objectives

Methodology

Results and Discussion

Anti-inflammatory Activity
(i) The Adjuvant Induced Polyarthritis Model

(ii) The Carrageenan Oedema Model

16

Correlation Between The Two Models

24

Carrier Studies
Carrageenan Induced Oedema

25

In Vivo Penetration Study

26

In Vitro Permeability Studies

27

Fatty Acid Profiles

28

Implications

29

Recommendations and Conclusions

31

References

37

25

RIRDC Publications

iv

Executive Summary
In the past, the profitability of the emu industry relied largely on breeding and selling chicks
into an expanding industry. However, it is apparent now that the future of the industry will
have to depend on the sale of emu products. The products include meat, skins, fat (oil) and
feathers. Based on the current and predicted prices for these products, it would seem likely
that emu farming will only remain profitable if the very high price previously paid for the
fat is maintained. Until recently, the price obtained for emu fat was between $20 and $25
per kilo, compared to around 20 cents per kilo for fat from other animal sources. However,
this has drop rapidly, as increasing supplies become available, and will continue to do so
unless high value end uses are established for the oil.
High prices are paid for emu fat because emu oil is believed by some to have therapeutic
and cosmetic applications. It is probable that in the near future the amount of emu oil
required for cosmetics production will be limited. Reasons for this include the small
percentage of oil in the products, the cost of production (around $2 per kilo for rendering
and around $5 per kilo for refining) and there is a lack of any documentary evidence that
emu oil performs any better than a range of cheaper alternatives. Furthermore, particularly
in Australia, there is a move away from cosmetics that contain animal products (personal
communication, David Stacy, Orion Laboratories, WA).
In Australia and many overseas countries it is an offense to make any therapeutic claims
about a product until its efficacy has been demonstrated and the product registered with the
relevant authority. Therefore to expand the therapeutic uses of emu oil, registration as an
active ingredient under the Therapeutic Goods Act is required. The first step towards
registration is to demonstrate the efficacy of emu oil in a scientifically acceptable manner
Prior to the commencement of this study, the evidence for the efficacy of emu oil as an antiinflammatory agent has been largely anecdotal, such as Australian aboriginals have used
emu oil for centuries to treat inflamed joints. The first accounts of the efficacy of this oil
were published in the mid 1800's[1]. However recently, a wide range of therapeutic
applications for the oil have been claimed in two United States patents [2,3]. Unfortunately,
no statistical evaluation of the results was presented in these patents.
Also, it is often stated that emu oils from different sources and refined by differing
processes differ widely in their therapeutic activity but no scientific studies have been
published that confirm this.
The objectives of this study were:
1.

By using an appropriate animal model(s), to determine if emu oil alone or in

combination with other agents is efficacious in the treatment of inflammation.

2.

Isolate the active agents in emu oil or develop a test to determine the level of
efficacy of emu oil samples.

3.

Conduct clinical studies on the efficacy of emu oil.

4.
4.1.
4.2.
4.3
5.

To investigate the effects of the following factors on the efficacy, yield and
properties (physical and chemical) of emu oil:
fat handling and storage procedures
fat rendering and refining regimes
oil storage procedures.
To prepare specifications for the production of emu oil to be used in antiinflammatory applications.

In this study, the anti-inflammatory activities of different preparations of emu oil have been
examined using an adjuvant induced polyarthritis in rats and a carrageenan oedema in rats.
Adjuvent induced polyarthritis results in a chronic systemic inflammation with multiple
joint involvement. Carrageenan induces an acute localised inflammation. Both these rat
models have been commonly used for the detection and development of clinically effective
anti-inflammatory drugs.
Various emu oil preparations both from farm reared and wild birds have been shown to be
highly efficacious in both models. For example in the polyarthritis model, emu oil
preparations have shown to equal or greater anti-inflammatory activity to Ibuprofen, one of
the most powerful non prescription anti-inflammatory available. Also in the carrageenan
induced oedema model, emu oil preparations have been shown to have anti-inflammatory to
prednisolone, a powerful steroid anti-inflammatory drug.
However a wide range in efficacy was observed. The reasons for this wide variability in
efficacy remain to be established but some factors have been identified. These include diet
and method of oil preparation. Also it was found that it is highly likely that two different
components of the oils are the active ingredients in the two models. To date attempts to
identify the active constituents have been unsuccessful.
None of the emu oil preparation have caused any adverse effects such as skin irritation.
Analytical studies showed that emu oil prepared from the fat of farm reared birds consisted
of a relatively simple mixture of a limited number of compounds. Also that a single
triglyceride constituted around 70% of the oil. The triglyceride is of a type not generally
found in any quantity in animal fats in that it has the same unsaturated fatty acid (oleic) at
each end and a saturated fatty acid (palmitic) in the middle. The high concentration of this
triglyceride is probably responsible for the unusual properties of emu oil.
Whilst the data is consistent with this triglyceride being the major constituent of oils
prepared from farm reared birds this does not appear to necessarily be the case for oils
prepared from the fat of wild birds particularly those feeding on native flora. The variation
in fatty acid profiles of oils prepared from the fat of wild birds is much greater than that
seen with oils prepared from the farmed birds. This variation may reflect varying diets
since birds shot close together have similar profiles. Oils have been prepared from the fat
of wild birds that were 30% omega 3 fatty acid whereas with farmed birds this fatty acid
constituted less than 1% of the oil.

vi

Whilst only briefly addressed in this study it, would seem likely that emu oil may be useful
as a trans dermal carrier of therapeutic agents, particularly of agents that are soluble in emu
oil.
Objectives 1 and 4 were examined in detail but because of the nature of the results and
reduction in funding it was not possible to meet the other objectives.
However following recommendations and conclusions are made related to Objectives 1 and
4:
1.
Emu oils with potent anti-inflammatory activity can be prepared
2.

Different components of the oil are active in the different rat models.

3.

Factors affecting efficacy include diet and rendering procedure.

4.

Marked differences were found between the oils prepared from the fat of wild birds
and farmed birds.

5.

Further investigation into the anti-inflammatory activity of emu oil should be

undertaken. These studies should include human clinical trials at the earliest
opportunity.

vii

Introduction
In the past, the profitability of the emu industry relied largely on breeding and selling chicks
into an expanding industry. However, it is apparent now that the future of the industry will
have to depend on the sale of emu products. The products include meat, skins, fat (oil) and
feathers. Based on the current and predicted prices for these products, it would seem likely
that emu farming will only remain profitable if the very high price previously paid for the
fat is maintained. Until recently, the price obtained for emu fat was between $20 and $25
per kilo, compared to around 20 cents per kilo for fat from other animal sources. However,
this has drop rapidly, as increasing supplies become available, and will continue to do so
unless high value end uses are established for the oil.
High prices are paid for emu fat because emu oil is believed by some to have therapeutic
and cosmetic applications. It is probable that in the near future the amount of emu oil
required for cosmetics production will be limited. Reasons for this include the small
percentage of oil in the products, the cost of production (around $2 per kilo for rendering
and around $5 per kilo for refining) and there is a lack of any documentary evidence that
emu oil performs any better than a range of cheaper alternatives. Furthermore, particularly
in Australia, there is a move away from cosmetics that contain animal products (personal
communication, David Stacy, Orion Laboratories, WA).
In Australia and many overseas countries it is an offense to make any therapeutic claims
about a product until its efficacy has been demonstrated and the product registered with the
relevant authority. Therefore to expand the therapeutic uses of emu oil, registration as an
active ingredient under the Therapeutic Goods Act is required. The first step towards
registration is to demonstrate the efficacy of emu oil in a scientifically acceptable manner.
Rural Industries Research and Development Corporation (RIRDC) invited Agriculture
Western Australia to facilitate a national emu oil research and development project over the
period 1996/97 to 1998/99. As part of the facilitation, Agriculture Western Australia
organised a meeting, sponsored by RIRDC, of industry representatives and researchers with
expertise and interest in emu oil research to develop a plan for a national emu oil research
development program to be funded by RIRDC. The participants identified the following as
areas requiring further research, the areas are listed in order of priority:
1. Anti-inflammatory properties of emu oil.
2. The effects of emu oil on wound healing and cellular regeneration.
3. Anti-viral/anti-bacterial activity.
4. Cholesterol lowering cardiovascular effects of emu oil.
Subsequently, RIRDC funded projects in the first three areas.
Prior to the commencement of this study, the evidence for the efficacy of emu oil as an antiinflammatory agent has been largely anecdotal, such as Australian aboriginals have used
emu oil for centuries to treat inflamed joints. The first accounts of the efficacy of this oil
were published in the mid 1800's[1]. However recently, a wide range of therapeutic
applications for the oil have been claimed in two United States patents [2,3]. Unfortunately,
no statistical evaluation of the results was presented in these patents.

Also, it is often stated that emu oils from different sources and refined by differing
processes differ widely in their therapeutic activity but no scientific studies have been
published that confirm this.
In this study, the anti-inflammatory activities of different preparations of emu oil have been
examined using an adjuvant induced polyarthritis in rats and a carrageenan oedema in rats.
Both these rat models have been commonly used for the detection and development of
clinically effective anti-inflammatory drugs [4-6].

Objectives
1.

By using an appropriate animal model(s), to determine if emu oil alone or in

combination with other agents is efficacious in the treatment of inflammation.

2.

Isolate the active agents in emu oil or develop a test to determine the level of
efficacy of emu oil samples.

3.

Conduct clinical studies on the efficacy of emu oil.

4.

To investigate the effects of the following factors on the efficacy, yield and
properties (physical and chemical) of emu oil:
4.1.
fat handling and storage procedures
4.2.
fat rendering and refining regimes
4.3
oil storage procedures.

5.

To prepare specifications for the production of emu oil to be used in antiinflammatory applications.

Methodology
Emu Oil Samples
The details of the oils used in a particular study are given in Results and Discussion.
Adjuvant Induced Polyarthritis
Adjuvant arthritis was induced by injecting a mixture of M.tuberculosis (800 ug) in
squalane (0.1mL) into the tail base of female Wistar rats (160-200g) on day 0. On day 10,
the rats were shaved just behind the ears to expose approximately 6 cm2 of dorsal skin.
Mixtures of 85% v/v emu oil and 15% cineole, a penetration enhancer [7] were prepared. A
mixture of 85% olive oil and 15% cineole was used as a negative control. Four to six
animals per group were used. The mixtures were applied at the rate of 2mL/kg on days 10,
11, 12 and 13. The rear paw diameters were measured on days 10 and 14 using a
micrometer.

Carrageenan Induced Oedema

The rats were shaved just behind the ears to expose approximately 6 cm2 of dorsal skin the
day before testing.
The animals were anaesthetised using Halothane/oxygen and 0.1mL of 1% lamda
carrageenan dissolved in physiological saline was injected into the right hand rear foot pad.
Then 1mL of oil or sorbolene (control) was applied to the animal, approximately 0.2 mL
rubbed into the affected paw and 0.8 mL rubbed into the shaved area. Eight animals per
group were used. The paw diameters were measured at 0, 4 and 6 hours.
The paws were also inspected at 24 hours and the results from any animals showing
heamorrhage at the injection site were rejected.
In vivo Penetration Study
A commercial emu oil preparation (EO1) and oil prepared from the intra-dominal fat of
birds raised by Agriculture WA (EO2) were used in this study.. Approximately 1 kg of fat
was heated in a 650 watt microwave oven on high for about 20 minutes. The oil was then
filtered through cotton wool, stored in a glass beaker sealed with plastic wrap.
Fifteen rats were anaesthetised with ether and shaved just behind the ears to expose
approximately 6 cm2 of dorsal skin. Blood samples, approximately 0.5mL, were then
obtained by heart puncture. 0.5mL samples of hydrocortisone (5mg/mL) dispersed in emu
oil or a commercial preparation of hydrocortisone (5mg/mL) cream were then applied to the
shaved area and rubbed in for 20 seconds. Further blood samples were taken at 3 and 5
hours. Prior to taking blood samples, the animals were anaesthetised. The blood was
allowed to clot and the serum collected. The serum samples were kept frozen until
required.

The hydrocortisone content of the serum was determined using a commercially available
radio immune assay ( Incstar Corporation, Stillwater, Minnesota,U.S.A.). The
measurements were performed by the Chemistry Centre (WA) on contract to Agriculture
Western Australia.
In vitro Permeability Studies
These studies were performed by InterDerm Pty Ltd on contract to Agriculture WA.
Briefly, the test procedure used glass diffusion cells with horizontally mounted sections
(approximately 1cm2) of human epidermal membrane from a single donor. Test
formulations composed of the radioactively labeled drug dispersed in the relevant oil were
applied to the upper stratum corneum side and the penetration of the labeled drug into the
underlying receptor medium was measured over a 48 hour period.
For the first experiment, H3-cortisone was the drug used. The cortisone was dispersed in
olive oil (control), commercial emu oil (EO1) or emu oil prepared by Agriculture Western
Australia (EO3). (EO4) was prepared by Dr C. Davis, Department of Primary Industries,
Hamiliton, Queensland. The oil was prepared from the subcutaneous fat of an emu raised at
Cherbourg. The oil was obtained by low temperature (40oC) rendering and clarified by
centrifugation at 12,000g for 10 minutes at 30oC. As tested, the oil was almost colourless
with a small quantity of solids at room temperature. The tissue sample was obtained from a
82 year old female.
The second experiment was essentially the same as the first except the drug used was C14diclofenac, a non steroid anti-inflammatory drug, and the tissue sample was obtained from
an 78 year old female.
The third experiment was essentially the same as the first except the hydrocortisone was
dispersed in EO2, and the tissue was obtained from an 25 year old female.
Fatty Acid Analysis
Routine fatty acid analyses were performed by the Animal Health Laboratories, Agriculture
Western .Australia using established techniques including Gas Chromatography (GC).
GC/Mass Spectroscopy and Nuclear Magnetic Resonance Studies
The CG/Mass Spectroscopy and Nuclear Magnetic Resonance Studies were performed by
the Centre for Drug Design, University of Queensland using oil samples supplied by
Agriculture Western .Australia.

Results and Discussion

Anti-inflammatory Activity
(i)The Adjuvant Induced Polyarthritis Model.
In the initial study, the anti- inflammatory properties of four different preparations of emu
oil were examined. These were sourced from birds raised in quite different habitats, three
of these were from Western Australia (EO1, EO2 and EO3) and one from Queensland
(EO4). Olive oil was used as the control.
It can seen ( Figure 1) that dermal application of three of the four emu oil preparations
significantly reduced the increase in paw diameter due to arthritic inflammation over the
treatment period. Also, EO4 was significantly more effective at reducing swelling than
EO2 or EO3, the negative value showing that it reduced incipient inflammation already preestablished on day 10. Anova of the mean paw measurements obtained in the oil study gave
a P of < 0.001

Figure 1. The effects of the various emu oil preparations on adjuvant induced arthritis.
None of the rats showed any adverse skin reactions at the site of application of any of the
oil preparations.
The administration of ibuprofen, one of the most effective anti-inflammatory drugs
available over the counter, significantly reduced (P<0.001) the increase in paw diameter due
to arthritic inflammation over the treatment period but on average it was less effective than
EO2, EO3 or EO4 (8).

The four oil samples where then subjected to a variety of analytical procedures, including
nuclear magnetic resonance (NMR) spectroscopy. Simplistically, the NMR spectroscopy
can be considered to give a fingerprint of the type of carbon and hydrogen atoms in the
material. A unexpected finding was the apparent simplicity of all the spectra (Figure 2.).
This indicates a relative simple mixture of a limited number of compounds.

Figure 2. A typical NMR spectrum of emu oil.

Also, there were no significant differences between the four spectra which indicates that the
active ingredient is only a (very) minor component of the oil. Using this and other data, the
major component (around 70%) of all the emu oil samples was identified as the triglyceride
shown in Figure 3.

OCO-(CH2)7-(CH=CH)-(CH2)7-CH3

CH3-(CH2)14-CO2

OCO-(CH2)7-(CH=CH)-(CH2)7-CH3

Figure3. The structure of the major component of emu oil.

This type of triglyceride is unusual to be in present animal and may, at least in part, be
responsible for the unusual qualities of emu oil in that there is a unsaturated fatty acid (oleic
acid) each end and a saturated fatty acid (palmitic acid) in the middle.

EMU OIL 5 (EO5) WAS A COMMERCIALLY RENDERED PREPARATION

PREPARED FROM THE FAT OF BIRDS RAISED AT CHERBOURG, QUEENSLAND.
AS SUPPLIED THE OIL CONTAINED APPROXIMATELY 25%(V/V) SOLIDS. THE
CRUDE OIL WAS FILTERED BEFORE USE. THE OIL USED WAS A PALE YELLOW
VISCOUS LIQUID.

To examine the dose response, mixtures of EO5 and olive oil in the ratios of 1:1 (EO:OO
1:1) and 1:3 (EO:OO 1:3) were prepared. Adjuvant arthritis was induced in a further 16
female rats and on day 10 the rats were shaved and the rear paw diameters measured as
described above. Mixtures of 85% EO5, EO:OO 1:1 or EO:OO 1:3 and 15% cineole were
prepared and each applied to four rats (2mL/kg) on days 10, 11, 12 and 13. Again a mixture
of 85% olive oil and 15% cineole was used as a negative control on four rats. The rear
diameters were measured again on day 14.
The dose response obtained is shown in Figure 4, relative swelling is the ratio of the
swelling in the emu oil treated animals to that in the olive oil treated animals.

Figure 4. Dose response observed with EO5.

The next four oils to be tested were prepared from the fat of two birds raised by Agriculture
Western Australia feed on the standard heat based diet. Oil samples EOA, OEB, EOC and
EOD were prepared by heating samples of diced fat, two thirds subcutaneous and one third
gut, in covered glass beakers at 50, 60, 80 or 100oC for 3 hours respectively. Once again a
range of efficacies was observed (Figure 5).

Figure 5. The effect of rendering temperature on efficacy. EOA was rendered at 50oC,
EOB at 80oC, EOC at 80oC and EOD at 100oC.
No simple correlation between efficacy and rendering temperature was observed.
To examine the effects of diet, twelve diets were prepared consisting of a common basal
diet to which was added different lipid (oils or fat) preparations. Each diet was fed to three
birds for twelve weeks prior to slaughter. The fat was collected at slaughter and stored
frozen until required.
In order to obtain larger quantities of oil an alternative rendering procedure was used. The
fat was minced and the oil extracted by placing the fat on wire mesh in an oven at varying
temperatures (60, 80 or 100oC) for varying times
For the first study the oil extracted at 60oC from the subcutaneous fat of birds fed on of four
different diets (A, B, C and D) were used. The diets were prepared by adding 10% of oil to
a wheat based basal diet. Diet A had cod liver oil , Diet B had a hydrogenated mixture of
canola and palm oils (40% trans 18:1), Diet C had tallow and Diet D had canola oil. The
results of the study are shown in Figure 6.

10

Figure6. The effects of diet emu oil efficacy.

The swelling of the animals treated with the emu oils from all diets was significantly less
than the controls. Furthermore there were significant differences between diets.
The oils tested in this model have been subjected to chemical analysis, including fatty acid
composition, and there appears to be a good correlation between efficacy and the
concentration of an unidentified compound (Figure 7), presumably a polyunsaturated long
chain fatty acid.

11

Figure7. The relationship between efficacy and the concentration of the unidentified
compound.
The highest concentration of this compound has been approximately 5mg/mL and so it is
not surprising that we were unable to detect differences between the NMR spectra of
efficacious and non-efficacious samples of oil. We have been unable to identify the
compound using GC-Mass spectroscopy.
There is considerable interest in the properties of oil prepared from the fat of wild birds.
Birds were shot in three distinct regions of Western Australia and oil samples were prepared
by rendering the fat in a water bath at 60 oC.
The first three samples to be tested were to have been prepared from the fat of birds
collected from 3 widely separated regions of Western Australia. The preparations were
supposed to prepared from WE1 from the fat of wild bird 2 (WB2) shot in the South West
forest region, WE2 from WBs5&6 shot at Southern Cross, a eucalyptus and mallee region
and WE3 from WBs 11&12 shot at Paynes Find, a mulga region. However, it is apparent
from subsequent fatty acid analysis that an error had been made and WE2 had been

12

prepared from the fat of WB1, shot in the South West region. It can be seen (Figure 8) that
only one preparation (WE3) significantly reduced swelling.

Figure 8. The efficacy of emu oil prepared from the fat of wild birds.
WB1 and WB2 were both females shot in the Augusta region of Western Australia. They
were grazing on pasture, mainly weeds, and both stomachs were packed with dandelion
flowers. WB 11 was a male and WB 12 was a female both were shot outside the dog fence
in the Mount Magnet area and their stomachs contained mulga and quandong seeds.

13

The fatty acid profiles obtained for these samples are very interesting for a number of
reasons. Firstly, WE1 and WE2 have little any efficacy and have very similar profiles,
which are markedly different from that obtained for the active WE3 (Table 1).
Furthermore, the fatty acid profiles obtained for the oils prepared from the wild birds are
very different from the profiles obtained for oils prepared from farmed birds. These
differences include:
1. The ratio of C181n9:C16. With all oils prepared from farm reared birds this ratio is
close to 2:1(Table 1) as it is with WE1 and WE2 the ratio is close to 2:1 but with WE3
the ratio is 6.6:1.
2. Twelve percent of WE3 is an unidentified fatty acid, running between C204n6 and
C205n3.
3. Almost 30% of WE1 and WE3 is C183n3 whereas with the farmed birds this was
generally less than 1%.

14

Table1. Fatty acid profiles of emu oils prepared from the fat of farm bred (FB) and wild
(WE) emus.
FBD1 FBD2 FBD3 FFBD4 FBD5 WE1
WE2
WE3
Fatty
R.T. %
%
%
%
%
%
%
%
Acid
C14:0
?
C15:0
?
C16:0*
?
?
C16:1n
7
C17:0
?
?
C18:0
?
C18:1n
9* CIS
?
?
?
C18:2n
6*
?
?
C18:3n
3*
?
?
?
?
?
C20:4n
6
?
?*
C20:5n
3
C24:0
C22:4n
6
?

2.34
2.66
2.86
3.21
3.38
3.73
3.68
3.78

0.40% 0.80% 0.50% 0.60% 0.50% 0.60%

0.10% 0.10% 0.10% 0.10% 0.10%
0.00% 0.10% 0.00% 0.10% 0.10% 0.10%
0.10%
23.10% 24.00% 21.60% 22.00% 29.10% 17.00%
0.10%
0.10% 0.40%

3.95
4.22
4.31
4.74
5.15
5.53
5.73

0.10%
0.20%
0.10% 0.20% 0.10% 0.20% 0.20%
0.20%
0.20% 1.60%
0.10%
8.50% 10.00% 8.80% 9.20% 12.10%
0.30% 0.30% 1.50%
0.30%
50.60% 48.10% 51.20% 51.50% 43.70%

0.60% 0.20%
0.10%
0.10% 0.10%
16.40% 8.70%
0.20%

5.10% 5.20% 4.60% 5.00% 3.60% 1.80% 1.70% 0.10%

0.20%

9.20%
0.20%
31.00%

0.90%
0.00% 0.20%
0.20%
0.10% 0.20%
8.80% 5.90%
0.10% 0.10%
29.90% 57.20%

5.83 2.40% 2.30% 2.50% 2.10% 1.90% 1.40% 1.30% 0.70%

6.14 0.10% 0.20% 0.20% 0.20% 0.20%
0.10%
6.4
0.10% 0.20%
6.61 6.60% 4.10% 6.40% 5.80% 0.20% 8.80% 9.00% 5.90%
7.41 1.30% 1.40% 1.30% 1.40% 1.80% 1.50% 1.60% 0.10%
7.88 0.10% 0.20% 0.10% 0.10%
7.96 0.80% 0.30% 0.50% 0.40% 0.10% 27.40% 29.20% 6.10%
8.03 0.50% 0.30% 0.10% 0.20% 0.30% 0.20% 0.20% 1.40%
8.5
1.30% 0.40% 0.40% 0.20%
9.22
0.50%
9.7
1.20%
10.31
11.47 0.00% 0.00% 0.10% 0.10% 0.00% 0.00% 0.00% 0.00%
11.57
0.50% 0.00%
0.40% 0.40%
12
11.90%
13.24 0.00% 0.10% 0.00% 0.00% 0.00% 0.10% 0.00% 0.00%
15.03 0.00% 0.00% 0.00% 0.00% 0.00% 0.00% 0.00% 0.00%
15.55 0.00% 0.00% 0.00% 0.00% 0.30% 0.00% 0.00% 0.00%
17.8

0.20% 0.20% 0.10%

15

D1 was a standard diet with no added fat or oil. D2, D3, D4 and D5 were a basal diet to
which 10% cod liver, hydrogenated canola/palm, tallow and canola oil was added
respectively.

16

Additional oil samples were prepared from the fat of individual wild birds. Unfortunately
there was some spillage during transport and to obtain sufficient material for testing the
samples had to be diluted 1:1 with olive oil prior to testing. Even so all oil preparations
were found to be highly efficacious (Figure 9).

Figure 9. The efficacy of oils prepared from the fat of individual wild birds.
No correlation has been found between the efficacy of the oils prepared from the fat of the
wilds and the fatty acid profiles of the oils (Figures 8&9, Table 2).
It is interesting to note that that the oils prepared from birds shot outside the verminfence,
and therefore living on native fauna and flora, are very much more efficacous than the oils
prepared from the wild birds feeding on improved pasture.

17

Table 2. The fatty acid profiles of oils prepared from the fat of wild emus.

Fatty Acid
C14:0
?
C15:0
?
C16:0*
?
?
C16:1n7
C17:0
?
?
C18:0*
?
C18:1n9
CIS*
?
?
?
C18:2n6
?
?
C18:3n3*
?
?
?
?
?
C20:4n6
?*
?
C20:5n3
C24:0
C22:4n6
?
C22:6n3
?
?

WB1 WB2 WB5 WB6 WB1 WB1 WB1 WB1

0
1
2
4
R.T.
%
%
%
%
%
%
%
%
2.34 0.8% 0.6%
2.66
2.86 0.1% 0.1%
3.21
3.38 15.8 16.5
%
%
3.73
3.68
3.78 1.8% 1.6%
3.95
4.22 0.2% 0.2%
4.31
0.1%
4.74
5.15 8.0% 9.0%
5.53 0.1% 0.1%
5.73 33.1 30.0
%
%
5.83 1.5% 1.3%
6.14 0.1%
6.40
6.61 11.7 9.1%
%
7.41 1.5% 1.6%
7.88 0.1%
7.96 24.0 28.9
%
%
8.03 0.3% 0.2%
8.50 0.1%
9.22
9.70
10.31
11.47 0.1% 0.4%
11.57 0.3%
12.00 0.2%
13.24 0.1% 0.1%
15.03 0.0% 0.0%
15.55 0.0% 0.0%
17.80 0.2% 0.3%
18.28 0.0% 0.0%
19.73
20.17

0.4% 0.3% 0.5% 0.2% 0.1% 0.2%

0.1%
0.0% 0.0% 0.1% 0.0% 0.0% 0.0%
25.0
%

20.0 17.4% 9.7%

%
0.1%
0.1%
6.1% 3.7% 2.8% 1.0%

7.2% 11.5%
0.2% 0.1%
0.1%
0.9% 2.4%

0.1% 0.0% 0.2% 0.2% 0.2% 0.2%

8.3%
0.2%
48.9
%
2.1%
0.2%

0.2% 0.3% 0.2%

8.7% 8.9% 7.0% 4.7% 7.3%
0.2% 0.1%
47.1 38.7% 55.9% 59.7% 58.7%
%
1.9% 1.7% 0.8% 0.6%
0.1% 0.1%
0.1%

4.2% 4.8% 15.8% 7.4% 4.3% 4.8%

0.1%
1.3% 1.4%
0.1%
1.7% 10.6 12.6% 7.4% 4.0% 4.6%
%
0.4% 1.2% 1.7% 1.3%
0.4% 0.4%
0.1%
0.2%
0.2%
0.0% 0.0% 0.1% 0.1% 0.0%
1.2% 0.8% 0.1% 8.5% 15.7%
0.0%
0.0% 0.0% 0.0% 0.0% 0.0%
0.0% 0.0%
0.0% 0.0%
0.0% 0.0% 0.0% 0.1% 0.1%
0.1% 0.1%
0.0% 0.0% 0.0% 0.0% 0.0%

18

0.1%
0.1%
8.1%
0.0%
0.0%
0.0%
0.1%
0.0%

19

(ii)The Carageennan Induced Oedema Model.

The second animal model used is referred to as the carrageenan induced rat paw oedema
model. Whilst progress with using the carrageenan induced rat paw oedema model to
demonstrate efficacy was slower than with the adjuvant model, the model is now better
understood. It is now possible to distinguish between the two effects of carrageenan i.e. the
rapidly induced oedema (0 to 6 hours) and the development of granuloma (1 to 5 days).
Initially, considerable time and effort were spent examining the effects of emu oil on
granuloma development with limited success.
With this model, a small quantity of carrageenan, a sulphated polysaccharide, is injected
into one of the rear paws of rats. The diameters of the rear paws were measured and the oil
preparations applied to a shaved area on the back of the rats. The paw diameters were
measured at 2, 4 and 6 hours. The degree of swelling is taken as a measure of the severity
of the disease.
For the initial study using the revised procedure four oils were prepared from the fat of two
birds raised by Agriculture Western Australia. Oil samples EOA, OEB, EOC and EOD
were prepared by heating samples of diced fat, two thirds subcutaneous and one third gut, in
covered glass beakers at 50, 60, 80 and 100oC for 3 hours respectively. A range of
efficacies was observed (Figure 10).

Figure 10. The effects of emu oil preparations on carrageenan induced oedema.

20

At 2 hours, all treated animals showed significantly less swelling than the controls. The
animals treated with EOA, EOC or EOD also showed significantly less swelling at 4 and 6
hours. The oils extracted at the higher temperatures (80 and 100oC) were found to be more
efficacious than those prepared at the lower temperatures.
As described previously, to examine the effects of diet, ten diets were prepared consisting
of a common basal diet to which was added different lipid (oils or fat) preparations. A
sample of fat from a Tasmanian bird, feeding on forage, was also included in the study. The
fat samples were rendered in an oven at varying temperatures (60, 80 or 100oC) for varying
times.
The oils extracted at 60oC from the subcutaneous fat of birds fed on diets A, B, C and D
were used. Diet A had cod liver oil as the added lipid, Diet B had hydrogenated fat, Diet C
had tallow and Diet D had canola oil.
The results obtain when the efficacy of these oil samples were tested using the oedema
model are show in Figure 11, eight animals per treatment were used.

21

Figure 11. The effect of diet on the efficacy of emu oil.

At 2 and 4 hours there was no significant difference between the results obtained with the
control (sorbolene treated) animals and those animals treated with the different emu oil
preparations. However, at 6 hours there was a slight but significant difference (P<0.02)
between the controls and the Diet B treated animals. This result was unusual in that in
previous studies the efficacy had shown up earlier. Also, the average swelling of the Diet D
treated animals was significantly greater than that of the Diet B treated animals at both 4
and 6 hours.
It was considered possible that the apparent lack of efficacy may have been a result of the
low yields of oil obtained with the 60oC oven temperature, only approximately 10 percent.
It was decided therefore to use the oil extracted from the same fat samples at 80oC where
the yield was around 80 percent. Essentially the same results were obtained (Fig. 12).

Figure 12.
That is no significant difference between the control and emu oil treated animals at 2 and 4
hours but a slight, but again significant (P= 0.03), difference between the control and Diet B

22

treated animals at 6 hours. Also, again the average swelling of the Diet D treated animals
was significantly greater than that of the Diet B treated animals at 2, 4 and 6 hours.
For the next study the oils prepared from the subcutaneous fat of birds fed on four different
diets (E, F, G and H) were used. Diet E was the basal diet with no added lipid, Diet F had
olive oil added, Diet G had linseed oil added and Diet H was the forage diet. The oils were
extracted in the oven at 800C. When tested using the rat oedema model, none of the oil
preparations were found to be efficacious (Figure 13).

Figure 13.
Since efficacious oils had been prepared previously from the fat of birds on essentially the
basal diet (Figure 10), it seemed likely that some other factor (factors) was responsible for
the absence of, or low, activity of these oil preparations. Two possibilities were the lack of
gut fat or the rendering procedure. The oils which had shown efficacy previously had been
prepared from a mixture composed of two thirds subcutaneous fat and one third gut fat. In
the latter preparations minced fat was placed on a stainless steel mesh and placed in an oven
at the required temperature. The oil dripped through the mesh and was collected in a
stainless steel container. Whilst the previous oils had been prepared by dicing the fat with a
knife, placing it in a glass beaker which was covered with plastic wrap to exclude air and
placed in a water bath at the required temperature.
For the next study, the four oil samples tested were two prepared using the oven from the
gut fat of birds fed Diet B or D, a commercially rendered oil from birds fed on essentially

23

the basal diet and a sample where Vitamin A (~ 1mg/mL) was added to the commercially
prepared oil. This last sample was included because it was claimed in a United States
patent that bleaching the oil destroyed its anti-inflammatory activity and the addition of
retinal acetate to the bleached oil restored its activity. The results of the study are shown in
Figure 14.

Figure 14.
There was no significant difference between the results obtained with the control animals
and those obtained with the animals treated with Diet B and D emu oils. The animals with
the commercially rendered oil had significantly (P=0.02) less swelling than the control
animals at 2 and 4 hours but not 6 hours. However, the addition of the Vitamin A resulted
in the swelling being the same as the control animals.
All oils described to this point have been chemically characterised but no correlation
between chemical composition and efficacy has been established.
At this time it was decided to discontinue testing oils prepared by heating in the oven and
go back to the original method of heating in sealed glass containers. Also, it was decided to
use samples composed of two thirds subcutaneous and one third gut fat.

The next study was a combination study. The oil used was prepared from the fat of birds
fed diet B (Diet B*) using the new rendering procedure. The other samples tested were this
oil to which various long chain polyunsaturated fatty acids were added at the concentrations
shown in Figure 15. The oil samples and sorbolene were only applied once, immediately
after the injection of the carrageenan.

24

Figure 15. Emu oil as a trans-dermal carrier.

The results obtained with animals treated with the oil alone (Figure 15) appears to very
different from that observed previously with the oil prepared from the same fat using the
oven method. In this study there was a significant (P<0.01) reduction in swelling at 2 hours
after which there appears to have been a rebound reaction which could be expected if the
level of the active ingredient had fallen below efficacious levels. At two hours there were
no significant differences between the animals treated with oil alone and those animals that
had been treated with oils that the various compounds had been added. At all times there
was no significant difference between the oil alone and the oil plus substance A ( cis
5,8,11,14,17 Eicosapentaenoic acid). However, the animals treated with oil plus substance
B ( Arachadonic acid) or C ( cis 11,14, 17 Eicosatrienoic acid) had significantly less
swelling than those treated with oil alone at 4 and 6 hours.

25

To test the rebound theory, the Diet B* oil was applied 0, 2 and 4 hours. The results show
(Figure 16) that in this case the oil resulted in a significant reduction in swelling at all times
and no rebound was observed. At the same time oils prepared from the fat of birds fed diet
A (Diet A*) or diet D (Diet D*) were tested together with the oil prepared from a
commercially bred emu (Prod.1). These oils were only applied once at time zero.

Figure 16.
The Diet D* and Prod.1 oils produced no significant reduction in swelling (Figure 16). The
animals treated with Diet A oil had significantly less swelling than the controls at 4 hours.
The Diet B* was significantly more effective than the other oil preparations so diet certainly
appears to influence efficacy.
It is interesting to compare the results of the effect of repeated application of the active oil
(Figure 17).

26

Figure 17. The effects of repeated application of an active oil.

There has been considerable interest in the properties of oil prepared from the fat of wild
birds. Birds were shot in three distinct regions of Western Australia and oil samples were
prepared by rendering the fat in a water bath at 60 or 80oC. The results of the first study
using these oils at 0 and at 2hours are shown in Figure 18.

27

Figure 18. The efficacy of emu oils prepared from the fat of wild birds.
At 2 hours the oil treated animals were the same and had markedly less swelling than the
controls. At 4 hours animals treated any one of three oil preparations were the same and
with considerable less swelling than the controls. At 6 hours three of the oil treatments
have the same swelling as the controls and there appears that there may have been some
rebound.

28

The first three samples to be tested were to have been prepared from the fat of birds
collected from 3 widely separated regions of Western Australia. The preparations were
supposed to prepared from WE1 from the fat of wild bird 2 (WB2) shot in the South West
forest region, WE2 from WBs5&6 shot at Southern Cross, a eucalyptus and mallee region
and WE3 from WBs 11&12 shot at Paynes Find, a mulga region. However, it is apparent
from subsequent fatty acid analysis that an error had been made and WE2 had been
prepared from the fat of WB1, shot in the South West region.
The rendering temperature (60 or 80oC) had no effect on the fatty acid composition, i.e.
WE3(60) and WE3(80) had almost identical fatty acid profiles.
The results obtained with additional samples prepared from the fat of individual wild birds
are shown in Figure 19.

Figure 19. The efficacy of emu oils prepared from the fat of wild emus.
All these samples prepared from the fat of the wild birds had similar efficacies but had
widely differing fatty acid profiles (Table 2).

29

The last two oils to be tested were prepared from the fats of WBs 11 and 12. The results
are shown in Figure 20.

Figure 20. The efficacy of emu oils prepared from the fat of wild emus.
At all times the average swelling of the emu oil treated animals was less than the controls,
the differences were not statistically different (P<0.05) from the controls.
EVEN WITH THE CURRENT RENDERING TECHNIQUE CONSIDERABLE
VARIATION IN THE EFFICACY IS OBSERVED. THE REASONS FOR THE
VARIATION REMAIN TO BE ESTABLISHED. TO DATE NO CORRELATION
BETWEEN CHEMICAL COMPOSITION AND EFFICACY HAS BEEN ESTABLISHED
FOR THIS MODEL.
The maximum reduction in swelling observed has been 40% of the paw thickness. This still
may represent considerable potency since published studies show that even a very powerful
anti-inflammatory drug such as hydrocortisone will not prevent swelling occurring and will
only reduce the swelling by a comparable amount. To examine this the effects of topical
application of an active emu oil (WB5) at 0 and 2 hours were compared to the effects of

30

interperitonial injection of 1, 2 or 4 mg of prednisolone at 0 hours. The results of the study

are shown Figure 21.

Figure 21. Comparison of the efficacies of prednisolone and emu oil.

At 2 hours, all 3 doses of prednisolone gave the same result indicating this is the maximum
reduction in swelling that can be achieved with this drug which is around 40%. Since the
emu oil used achieved a comparable reduction in swelling to prednisolone, it must be
considered to have potent anti-inflammatory activity. The time course observed with the
prednisolone is different to that observed with the oil which suggests they have different
modes of action.
Correlation Between The Two Models
In Figure 22 the relative swellings (swelling in the treated animals/ swelling in the controls)
measured in the adjuvant model is plotted against that measured in the carrageenan model
when the same oil was tested in both models.

31

Figure 22. Correlation of emu oil efficacy.

It is apparent that there is little if any correlation between efficacy in the different models
Carrier Studies
Whilst not scientifically established, emu oil appears to penetrate the dermis much more
readily and more completely than other oils. If this is correct then emu oil may be useful as
a trans-dermal carrier of therapeutics agents.
Carrageenan Induced Oedema
Early attempts to use emu oil as a carrier for traditional anti-inflammatory drugs, aspirin,
phenylbutazone and hydrocortisone, were unsuccessful but in hindsight this was to be
expected, as they are not soluble in the oil. It was concluded that it would be more
appropriate to test materials that were soluble in emu oil.
Previous studies (Figure 15) had shown that when two substances (B and C) where added to
the emu oil (2.5 mg/mL) they appeared to significantly enhance the efficacy of the oil. So a
dose response study using substance B was undertaken. The concentrations used were 5,
2.5 or 1.25mg/mL. In this test, the carrageenan produced a very severe inflammation in
both the test and control animals. The inflammation was very much more severe than seen
normally and none of the applied materials significantly effected the severity of the
inflammation. The severity was such that it was considered it could be masking any
efficacy the material may have. The concentration of the carrageenan solution was checked

32

and found to be correct and there was no obvious explanation for the severity of the
oedema. The experiment was therefore essentially repeated but one group of animals was
given a high dose (10mg) of Ibuprofen. Again, very severe inflammation occurred in both
the test and control animals and none the of treatments, including the Ibuprofen, reduced
oedema. On further investigation, it was found that the carrageenan had been microwaved,
rather than, autoclaved into solution. A futher study confirmed microwaved carrageenan
produced a more severe oedema than the autoclaved carrageenan.
We have been unable to repeat this dose response study.
In the light of this, the results of previous studies (e.g. Figure 13) where very severe
inflammation occurred will have to be re-assessed.
It was considered that because of its structure, Ibuprofen should have reason solubility in
emu oil. For the final study in this series the efficacy of an oil prepared by commercially
rendering fat from birds raised by Agriculture (Commercial) with that of the oil to which
Ibuprofen (10mg/ml) had been added. For comparison the oil from WB12 was included.
(Figure 23).

Figure 23. Emu oil as a trans-dermal carrier of Ibuprofen

33

At all times the average swelling of the oil treated animals was less than the controls, the
only statistically significant difference was between the animals treated with the oil
containing Ibuprofen and the controls at 2 hours.

In Vivo Penetration Study

The average hydrocortisone levels measured in the blood of animal treated with
hydrocortisone dispersed in a commercially available emu oil (HCEO1), an oil prepared
from intra-dominal fat (HCEO2) or a commercial hydrocortisone (HCCOM) cream are
shown in Figure 24.

Figure 24.
The results showed that at two hours the level of hydrocortisone in the animal treated with
HCEO2 was significantly higher (P<0.05) than that of the animals treated with HCEO1 but
not significantly higher than those treated with the commercial cortisone preparation. There
was no significant difference the animals treated with HCEO1 and those treated with the
commercial cortisone.

34

At 5 hours there were no significant differences in the concentrations of hydrocortisone.

The apparently significant finding has to be viewed with caution since in all cases the levels
of hydrocortisone are low, most at the limit of detection, and would represent only a very
slight contamination with the applied material. For example the highest concentration in
blood measured was a 100, 000 fold lower than the applied concentration. Furthermore, the
variance between replicates was very high.
It was decided to discontinue this form of study for a number of reasons including cost and
the trauma caused to the animals.
In Vitro Permeability Studies
The results obtained showed there was no significant differences between the steady fluxes
of hydrocortisone from the various preparations. The fluxes measured (fraction
applied/hr/cm2 SEM) in the first experiment were for hydrocortisone in EO1 1.88x104
4.1x10-5, for hydrocortisone in EO3 2.36x10-44.7x10-5 and for hydrocortisone in olive
oil 2.61x10-44.1x10-5. The fluxes measured (fraction applied/hr/cm2 SEM) in the third
experiment were for hydrocortisone in EO2 10.0x10-410x10-5 and for hydrocortisone in
olive oil 9.0x10-44x10-5.
The results also showed there was no significant differences between the steady fluxes of
diclofenac from the various preparations. The fluxes measured (fraction applied/hr/cm2
SEM) in the first experiment were for diclofenac in EO1 1.91x10-52.2x10-6 for diclofenac
in EO3 1.82x10-51.6x10-6 and for diclofenac in olive oil 1.64x10-52.6x10-6.
Fatty Acid Profiles
The fatty acids profiles of oil samples prepared from the fat of 8 wild birds are shown in
Table 2.
The main features to note are:
1. The wide variation in the fatty acid profiles. As stated previously, the variation seen in
fatty acid profiles of the oils prepared from the fat of wild birds vary much more than
that seen with farmed birds feed a variety of diets. This variation may reflect varying
diets because birds shot close together ((1&2), (5&6) and (11&12)) have similar
profiles.
2. As discussed previously, the omega 3 fatty acid content
3. Also as discussed previously, the C18ln9:C16 ratio.
4. The actual C19ln9 content. In farmed birds the C18ln9 content varied little (50+5%)
but with the wild birds this varies from 30 to 60%.
5. The variation in the amount of the unidentified fatty acid, running between C204n6 and
C205n3. Samples containing this fatty acid were sent to the Centre for Drug Design and
Development, University of Queensland for analysis by GC/Mass spectroscopy but the
substance was not identified.

35

Fatty acid analysis of the same oil preparation immediately after preparation and again after
storage at 5 oC showed that changes in fatty acid profiles did occur during storage. In
particular it was noted that the fatty acid that appeared to correlate with the efficacy of the
oils prepared from farm reared birds (see Figure 7) appeared to decrease during storage,

36

Implications
The demonstration that oil preparations with potent anti-inflammatory active can be
prepared and that there were no adverse effects of emu oil observed in this study nor in a
previous study (9) should facilitate obtaining permission from the relevant Ethics
Committees to perform human clinical trials on the anti-inflammatory activity of emu oil.
Such trials are required before Therapeutic Goods Act registration can be obtained.

37

38

Recommendations and Conclusions

Objectives 1 and 4 were examined in detail but because of the nature of the results and
reduction in funding it was not possible to meet the other objectives:
However the following conclusions and recommendations are given.
1.

Emu oils with potent anti-inflammatory activity can be prepared

2.

Different components of the oil are active in the different rat models.

3.

Factors affecting efficacy include diet and rendering procedure.

4.

Marked differences were found between the fatty acid profiles of oils prepared from
the fat of wild birds and those prepared from the fat of farmed birds.

5.

Further investigation into the anti-inflammatory activity of emu oil should be

undertaken. These studies should include human clinical trials at the earliest
opportunity.

39

40

41

References
1.
Bennett, G. Observations principally on the animal and vegetable products of
New South Wales, Gatherings of a naturalist in Australasia. London: John Van
Voorst, Paternoster Row: 1860.
2.

Ghosh P., Whitehouse M., Dawson M. and Turner A. Anti-inflammatory

composition derived from emu oil. United States patent number 5,431,924. 1995.

3.Fein E., Caputo J, and Nagal K. Therapeutic uses of emu oil. United States patent
number 5,472,713. 1995.
4.
Whitehouse, M. W. Adjuvant induced polyarthritis in rats. In: Greenwald
RA, Diamond HS, eds. Handbook of models for rheumatic diseases. Vol. 1. Boca
Raton: CRC Press: 1988:3-16.
5.

Rainsford, K. D. Adjuvant polyarthritis in rats: is this a satisfactory model for

screening anti-arthritic drugs? Agents Actions 1982; 12: 452-5.

6.

Billingham, M. E. J. Models for arthritis and the search for anti-arthritic drugs.
Pharmac Ther. 1983; 21: 389-417.

7.

Hashida M. Yamashita F. Terpenes as penetration enhancers. In: Smith EW

Maibach MI, eds. Percutaneous penetration enhancers. Boca Raton: CRC
Press: 1995:309-21.

8.

Snowden, J.M. an. Whitehouse, M.W. Anti-inflammatory activity of emu oils in

rats. Inflammopharmacology, 1977; 5, 127-132.

9.

Snowden, J.M, Roberts, M. and Cross, S. Emu oil in wound healing and cellular
regeneration. RIRDC Publication No. 98/18.

42