The opportunity for modern uses of indigenous and aboriginal medical practices

The opportunity for modern uses of indigenous and aboriginal medical practices

Amy Mann

Turramurra High School

Abstract

Modern medicine can be further advanced with the use of aboriginal practices. Dodonaea Viscosa, or otherwise known as hopbush, is a plant used in indigenous practices for antimicrobial activity. This experiment investigated the effect of the subspecies Dodonaea Viscosa Purpurea on the three bacterias, M.luteus, B.Subtilis, and E.coli. The bacteria was produced into pure cultures using the streak method in order to isolate the bacteria. Then the plant was produced into a liquid with incorporation of aboriginal practices, using saliva and a mortar and pestle to simulate chewing the plant. Then the liquid was tested against the bacterias with a disk diffusion method. The results showed that there was a large difference between the means of the zone of inhibition and the expected zone of inhibition. Which showed that the Dodonaea Viscosa Purpurea liquid has a very insignificant effect on the bacteria. This demonstrated that the subspecies has very little to no antimicrobial activity

Literature review

For tens of thousands of years, Aboriginal people have relied on natural remedies to treat a variety of illnesses, and thankfully, many of those remedies are still being used today (NACC, 2016). Australian bush medicine is a widely known traditional method for pharmaceutical practices that still hold many mysteries, even today (Kamenev, 2011).“[In Aboriginal culture], nothing is written down; instead, it’s passed on through singing and dancing ceremonies, which are becoming increasingly rare.” says Dr Evelin Tiralongo a pharmacist and expert in complementary medicine from Griffith University in Queensland(Australian Geographic, 2011). Different parts of Australia varied in the specific plants and food they’d eat, (Gott, 2018), which resulted in different plants and methods used for medicinal practice. The premature deaths of the custodians of Yaegl traditional knowledge meant this community lost much of its cultural memory. It was a personal tragedy but also a cultural, economic and intellectual one. There is great urgency about protecting what customary knowledge remains and using it to benefit communities (Tressider, 2019). Modern Medicine is becoming more reliant on plants, so being able to utilise the chemical and biological properties of medicinal plants should be a priority. Associate Professor of Molecular Chemistry Joanne Jamie is the Co-Director of Macquarie’s Indigenous Bioresources Research Group, along with medical microbiologist Associate Professor Subramanyam Vemulpad. These two are working to bridge the gap between bush and modern medicine. They have been working directly with the [Yaegl] community to document and preserve customary indigenous knowledge, both traditional and contemporary (Tressider, 2019). To address the dwindling information of bush medicine from indigenous people, Jamie.j et al (2022) aimed to investigate, with a literature review, the aboriginal medicinal plants native to Queensland, and research their uses and biodiversity. The research showed that there were 135 species of Queensland’s native indigenous plants that were used to treat 62 different skin diseases (Jamie.j, 2022). Additionally, the article demonstrated that the distinct cultures and beliefs that have come because of the aboriginal people show an intimate and profound relationship that they have developed with the environment (Jamie.j, 2022). These medicinal plants were said to have been rarely researched through western protocols, which shows a potential for the prospects of bush medicine aiding modern medicine (Jamie.j, 2022). Within this article, it can be seen that indigenous biocultural knowledge (IBK) is gaining an increasing importance and recognition for its possible value used within pharmaceuticals, biodiversity conservation, land management, and biodiscovery (Jamie.j, 2022). Similarly to the last article, in studying the methods and functions of bush medicines, Hossain(2019) investigated the uses of sticky hopbush, dodonaea viscosa, as a cure for a variety of ailments within the Omani nation. Utilising the knowledge from different pharmacological studies that show that extracts of D.visoca has significant antidiabetic, antimicrobial, insecticidal, antioxidant, cytotoxic, antifertility, anti-inflammatory, analgesic, anti-ulcer, antispasmodic, anti-diarrheal and detoxification effects (Hossain, 2019). D.viscosa is a species of plant belonging to the sapindaceae family, which has other species within the family similarly used in the treatment of a wide variety of illnesses (Hossain, 2019). Almost all parts and all species belonging to this family are used as a traditional medicine (Hossain, 2019). Most of them are flowering and woody plants and used medicine to treat different diseases including cardiac diseases (Hossain, 2019). African people used the plant's leaves to treat gastrointestinal disorders (GI disorders), skin conditions, and rheumatism (Hossain, 2019). In New Zealand, the leaves are used to treat wounds. However, in Oman the leaves of the selected plant as a medication to treat many ailments such as itching and rash, swelling, rheumatoid arthritis, bone disorders, GI disorder and the muscle relaxant (Hossain, 2019). While in Australia, people have used the plant to treat wounds, bleeding, bone fractures and snake bites (Hossain, 2019). This utilisation of the plant within folk medicines further highlights the antimicrobial and antiviral properties that the Omani considered of the plant (Hossain, 2019). After the investigations of Dodonaea Viscosa that reveal its medicinal properties, Nayeem.N (2021) further examined the plant by testing its wound healing properties on animal models. The experiment involved the use of dried leaves from the plant that was pulped in methanol and chloroform solvents to obtain a respective solvent of the extract (Nayeem.N, 2021). The wound healing activity of the herbal solution was then studied through Sprague Dawley rats using incision and excision wound models (Nayeem.N, 2021). The process of healing of a ruptured area holds complications that may eventually result in the restructuring and restoring of the original living tissue (Nayeem.N, 2021). The healing of wounds involves several stages that include inflammatory processes, contraction of wounds, and lastly epithelization (Nayeem.N, 2021). Agents, such as this dodonaea viscosa extract, can relieve this pain and work to enhance the properties that help in injury healing (Nayeem.N, 2021). From the study it was noticed that the methanolic extract formula diffused easier than the chloroform extract, however the difference was not significant (Nayeem.N, 2021). But with the diffusion characteristic in both the formulations, they demonstrated their significant wound healing activity in excision and incision wound models because it indicated that the formulation can pass through the medium to the site of action effectively (Nayeem.N, 2021). This research has shown that the dodonaea viscosa and other medicinal plants should be further researched and applied into the medical field. As several other natural substances have also been reported to expedite the healing process of a wound, these novel biodegradable, biocompatible, and economical components need to be identified for the healing of wounds.

Scientific research question

What are the effects of a Dodonaea Viscosa Purpurea medicinal liquid on the bacteria in the modern world?

Sub questions

What are the effects of a Dodonaea Viscosa Purpurea medicinal liquid on E.coli?

What are the effects of a Dodonaea Viscosa Purpurea medicinal liquid on B.subtilis?

What are the effects of a Dodonaea Viscosa Purpurea medicinal liquid on M.Luteus?

Scientific hypothesis

Alternative Hypothesis (H_a)

The Dodonaea Viscosa Purpurea Formulation will disrupt and prevent the bacteria from growing and produce an inhibition zone.

Null Hypothesis (H_0)

Bacteria and pathogenic agents are not affected by the medicine produced from the Dodonaea Viscosa Purpurea

Methodology

Procedure 1:

Production of pure cultures

Due to the safety protocols in school, the bacteria had to be produced from a pure bacteria culture rather than the dirt and water solution as thought before. There were three pure bacteria cultures produced, Escherichia coli (E.Coli), Bacillus (B.Subtilis) and Micrococcus (M.Luteus). These were used to create isolated bacteria colonies, which would then be used later in testing the antiseptic juice of the dodonaea viscosa.

bacterial solutions

1. Sterilise inoculating loop in bunsen burner

2. Wait for cool down

3. Dip loop into bacterial solution

4. Make a series of parallel non overlapping streaks on the edge of an agar plate

5. Sterilise inoculating loop in bunsen burner

6. Wait for cool down

7. Rotate agar plate 30°

8. Make a series of parallel non overlapping streaks starting in previously streaked plate

9. Repeat step 5-8 3 additional times

10. On the last streak, zigzag solution into middle of plate without touching the solution from other streaks

11. Close agar plate

12. Tape edges and label plate with bacteria

13. This is called the streak method and was used for all three of the bacterias.

Procedure 2:

Production of medicinal liquid

1. Extract 5 average sized leaves (7.5 ∓ 0.5 cm long and 1.3 ∓ 0.2 cm across) from the dodonaea viscosa, cutting them off the plant rather than pulling

2. Place these leaves into the mortar

3. Drop in 2 ml of saliva into the mortar

4. Grind leaves with a pestle in a circular motion for 3 minutes

5. Add another 2 ml of saliva and grind for another 2 minutes or until the leaves are dissolved but the stem is not

This should produce a goopy green mixture which will be used as the medicine.

Procedure 3:

1. How this medicine will be applied to the bacteria,

2. Light a bunsen burner

3. Open a pure bacteria culture

4. Gently pick up a small amount of the pure bacteria with a sterile cotton swab with the pure bacteria culture next to the flame but not touching

5. Then close the culture

6. Dip Cotton swab into a saline solution in a test tube

7. Dispose of cotton swab using methanol or alcohol

8. Gently mix saline + bacteria solution without spilling anything out

9. Pour solution into agar plate

10. Use a pipette to pick up the solution and place back into same test tube

11. Place plate down to let the leftover liquid settle into the agar for 10 seconds

12. Using a sterilised hole puncher, cut filter paper into 6mm diameter discs

13. Using a sterile tweezer, individually dip a 6mm filter paper disc into the medicine

14. Submerge disc for a second

15. Open up the agar plate

16. Place this individual onto one of the three bacteria cultures in the positions in figure 1. Below

17. Close the agar plate

18. Repeat steps 12 to 16 until all positions on the figure 1. are filled

Possible Errors

Table 2: Errors

Results

The results are based upon the facts. This section describes what was observed, calculated or the trends discovered. It is not an explanation of the results. The order of the results can either follow the order of the methodology or, maybe, in order of most important to least important. Results may include tables, graphs and/or other visual representations to highlight important features. It may be relevant to comment on the degree of uncertainty stated for each set of data collected. All visual displays should be labelled with a number, concise name and a stand-alone description of how the result was obtained. It is useful to integrate visual displays with text so that the reader is guided through the research.

Table 3: Bacteria zone of inhibition

Note: the purple highlighted data is the outliers within the data spread

To calculate whether the data is considered an outlier, utilising the Interquartile range to calculate.

Table 4: Data calculations

Figure 2: Average ZOI including outliers

Figure 3: Average ZOI excluing outliers

For this trial of this experiment, the standard zone is 6mm diameter circle with an area of 9π, and so for the zone of inhibition to prove the bacteria is sensitive to the dodonaea viscosa purpurea the zone of inhibition should reach 9π or greater when excluding the area of the standard zone.

Equation for working out area of zone of inhibition: A = πr^2 - 9π

Where r =3+ ZOI

Table 5: Bacteria resistance without outliers

From these results, it is seen that all three bacteria are resistant to the medicine, however the most susceptible bacteria was Bacillus Subtilis.

Statistical test includes an independent Ttest

Alpha-Value - 0.001: The alpha value, or the threshold for statistical significance, is arbitrary. In most cases, researchers use an alpha of 0.001, which means that there is a less than 0.1% chance that the data being tested could have occurred under the null hypothesis. The reason for the alpha value being 0.001 is from the article, Wound healing potential of Dodonaea viscosa extract formulation in experimental animals, (2021, Nayeem.N)

For the test to occur, I need to compare each bacteria to the ZOI that would be significant enough to be considered sensitive to the medicine. To prove the bacteria is sensitive to the dodonaea viscosa purpurea the zone of inhibition should reach 9π or greater when excluding the area of the standard zone.

Table 6: Bacteria resistance

Thus the radius of each the bacteria should of been:

9π = πr^2 - 9π

r^2 = 18

r = 3 + ZOI

3 + ZOI = 18

ZOI = 15

Therefore the ZOI must have been 15mm for the bacteria to be considered sensitive and have a significant effect.

Table 7: Expected value compared to actual value

Table 8: T-test

Since the p-value of the experiment shows that it is greater than the alpha-value, the null hypothesis is then accepted. Thus these bacteria and pathogenic agents are not affected by the medicine produced from dodonaea viscosa purpurea.

Discussion

Indigenous and Aboriginal medical practices have the ability to further develop modern medicine.

The Dodonaea Viscosa, hop bush, was used in order to determine whether it had an antimicrobial effect on the three bacterias, E.Coli, B.Subtilis and M.Luteus. The results from the experiment demonstrate that the two, E.coli and M.Luteus, are resistant to the medicinal liquid while B.Subtilis is also resistant but the most susceptible out of the three. This provides evidence to the null hypothesis which states that bacteria and pathogenic agents are not affected by the medicine produced from the Dodonaea Viscosa Purpurea. The main factor that demonstrates the very insignificant effect from the Dodonaea Viscosa Purpurea liquid was the difference of the means and the expected value. For the Dodonaea Viscosa Purpurea medicinal liquid to have been effective the means of the zone of inhibition would of had to have been 15 mm in radius, however the largest average zone of inhibition was for the Micrococcus luteus which had an average radius of 0.93mm. This provided evidence for the null hypothesis.

When broken down into its components, Dodonaea Viscosa, has fatty acids and starch that get broken down by the saliva because of amylase, lipase and other enzymes. These chemical compounds become antimicrobial and have a notable activity against different isolated human pathogenic bacterial strains, gram positive (2019, Hossain). This can be seen in the results as Micrococcus Luteus and Bacillus Subtilis are both gram positive and have a higher mean when including outliers. And then Escherichia Coli has the lowest mean and is a gram negative bacteria. Thus the medicine shows to be slightly more impactful on gram positive bacteria then gram negative but they still had a very insignificant effect from the medicine.

For this experiment, the subspecies of Dodonaea Viscosa Purpurea was used as it was accessible. This variety is a modern version of other traditional subspecies and was bred for looks and aesthetics for gardens. Previous research has utilised different subspecies from Saudi Arabia (2021, Nayeem.N) in which the most common variety is Dodonaea Viscosa Angustifolia (2020,Al-Namazi.A, Al-Ammari.B, Davy.A, Al-Turki.T). Another article utilised hop bush to research its biological and phytochemicals compounds (2019, Hossain). It demonstrates the pharmacological activities of the plant in antidiabetic, antioxidant, antimicrobial and other activities (2019, Hossain). In which the review was conducted with a Dodonaea Viscosa located in Omani, which commonly has the subspecies, Dodonaea Viscosa Angustifolia, like in Saudi Arabia. By using a different, under researched subspecies, it limits the chance for the Purpurea variety to hold a significant antimicrobial effect. All other assortments of Dodonaea Viscosa have been a part of folklore and traditional medicines, except for the Dodonaea Viscosa Purpurea. This is a possible reason as to why the medicinal liquid provided a statistical value (p>0.001) that demonstrates that the plant didn’t affect the bacteria significantly. While this variety of the plant shows an insignificant result, the use of other varieties, like Angustifolia, can be used to advance the research of medicinal plants and traditional medicines. This research may provide a negative result but aids in identifying plants that have medicinal properties. The method is limited in its conduction as the use of human saliva is a differing variable. In aboriginal practices, chewing the Dodonaea Viscosa was used and thus the saliva and plant mixed together (MDNC, 2020). This means that the effectiveness of the plant could depend on the saliva of the individual. This error poses a limitation to the aspects of antiseptic medicine produced and shows a degree of uncertainty in the plant. Another possible limitation is the subspecies.

By researching medicinal plants, the advancements of medicine can be enhanced with traditional practices. Indigenous medical practices can suggest plants and methods that produce sufficient medicinal application in modern health. The techniques used by aboriginal folklore give insight into the methods that can be used in medicine. Modern medicine relies on so many different chemical compounds and overcomplicates the production of medicine. Even with the complication of these medicines, bacteria evolve constantly and have been able to overcome the use of certain medications. Because of this, research should lean towards traditional practices to advance upon their research and create new alternate medicines that have the ability to overcome the evolution of bacterias.

Future research should focus on the utilisation of traditional medicine and practices to aid the advancements of medicine. Plants and herbal medicines have shown to be effective in cooperation with modern solutions.

Conclusion

This research was set in place to investigate the effects of a Dodonaea Viscosa Purpurea medicinal liquid on the bacteria in the modern world. The Dodonaea Viscosa Purpurea medicinal liquid was produced by incorporating traditional medicine practices. The medicine was tested against three different bacterial species, Micrococcus Luteus, Bacillus Subtilis, and Escherichia coli. The results showed that there was overall an insignificant antimicrobial effect from the Dodonaea Viscosa Purpurea liquid. This was demonstrated in the large difference between the means of the zones of inhibition of the bacteria and the expected value to be considered effective. Which meant that the research provided evidence for the null hypothesis which states that bacteria and pathogenic agents are not affected by the medicine produced from the Dodonaea Viscosa Purpurea The main limitation of this experiment is the subspecies used in this research. Other previous research has used other subspecies, mainly Dodonaea Viscosa Angustifolia, which did show to have a significant antibacterial effect. It was also seen that Dodonaea Viscosa Angustifolia was the subspecies that worked best within wound healing which includes antimicrobial activity. This research demonstrates that the use of aboriginal practices can provide some impact on modern medicine. Previous research provides a greater understanding of the mesh between folklore and modern medicine but this experiment provides information on the differences in subspecies.

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