IF plant extracts contain antiviral properties, THEN they will hold back the growth of viral-like activity in the model, BECAUSE these compounds have been shown to interfere with viral replication and are also able to destroy the integrity of viral particles.

 

ANTIVIRAL AGENTS IN PLANTS

Which plant(s) have antiviral properties, what are they?

For a long time plants have been used as a form of natural treatment against many illnesses, including viral infections. Some of the most common herbs used are : oregano, garlic, rosemary, ginger, ginseng and echinacea. These can be easily implemented into food, while also helping to protect you against many different viruses. For example, the Oleanolic acid in rosemary has shown inhibiting effects against herpes, HIV, influenza and hepatitis in test-tube and animal studies. Compounds found in other plants also have antiviral effects, such as :  

Oregano : Contains carvacrol and thymol, which have demonstrated antiviral effects against murine norovirus (MNV), herpes simplex virus type-1 (HSV-1), rotavirus, and respiratory syncytial virus (RSV).

Garlic : Rich in allicin, ajoene, and quercetin, which exhibits antiviral effects against various viruses including influenza A and B, HIV, HSV-1, viral pneumonia, and rhinovirus, which causes the common cold.

Rosemary : Contains oleanolic acid, which has presented antimicrobial activity against herpes, HIV influenza, and hepatitis

Ginger : Contains compounds like gingerol, shogaol, zingerone which exhibit antibacterial effects against avian influenza, RSV, and feline calicivirus (FCV)

Ginseng : Composed of ginsenosides, which have shown antiviral effects against hepatitis B, norovirus, and coxsackieviruses, as well as an infection of the brain called meningoencephalitis.

Echinacea : Contains alkamides, caffeic acid derivatives, and polysaccharides, which have demonstrated to inhibit viruses such as herpes and influenza.

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How do different plant extracts compare in their antiviral effectiveness? 

Plant extracts have been widely studied for their antiviral properties, with some showing strong activity against various viruses. Their effectiveness depends on factors such as the type of virus, active compounds, and how the extract is prepared. Some work by blocking virus entry, others inhibit viral replication, and some boost the immune system  to help fight infections.

1. Mechanisms of Antiviral Action 

Plant extracts can fight viruses in different ways: 

• Blocking virus entry : Some prevent viruses from attaching to or entering host cells. An example being oregano (Origanum vulgare)  contains carvacrol, which disrupts viral membranes, making it harder for the virus to infect cells. 
• Inhibiting Viral Replication : Certain compounds interfere with how viruses copy themselves inside the host cells. For example Rosemary (Rosmarinus officinalis) contains rosmarinic acid, which inhibits viral enzymes, slowing down replication. 
• Boosting the Immune System : Some extracts stimulate the immune response, helping the body fight infections more effectively. An example being Echinacea (Echinacea purpurea) enhances immune cell activity, making it useful to prevent colds and flu. 

2. Differences in Effectiveness 

Not all plant extracts work in the same way, and their effectiveness can vary based on : 

Broad-Spectrum vs. Specific Action 

• Garlic and Oregano have broad spectrum antiviral properties, meaning they can fight multiple types of viruses
• Ginseng and Echinacea are more specialized, mainly helping with viruses like influenza RSV.

Concentration and Preparation : 

• Fresh, dried, or alcohol-extracted forms can have different potency levels.
• Some compounds are heat-sensitive, meaning boiling them may reduce their efficiency.

Toxicity and Side Effects : 

• While generally safe, high doses of some extracts (like ginseng) can have side effects such as digestive issues or allergic reactions

3.  Antiviral Properties of Selected Plant Extracts 

 

4. Comparison of Antiviral Effectiveness 

 

5. Key Takeaways 

• Oregano and Garlic have strong direct antiviral effects, making them useful for fighting infections. They also stand out for broad-spectrum antiviral activity, meaning they work against many different viruses or infections.
• Echinacea and Ginseng are best for boosting the immune system response, helping the body fight viruses.
• Ginger and Rosemary work well against respiratory infections and may reduce symptoms of colds and flu.

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Does the extraction method affect antiviral activity?

Components in plant extracts are affected by factors such as temperature, pressure, environment, and method of extraction. Differences of what content is obtained by each extraction method is shown, its biological activities fluctuating. Simplified: yes, the extraction method does happen to change characteristics in plant extracts (e.g, hydroalcoholic extraction is better for clinical use).

Examples of this are present studies: different methods are shown to combat specific viruses and bacteria. CLSE (conventional liquid-solid extraction) agar method was shown to have weaker activity in inhibiting bacterial growth when testing ground peony flowers. MAE (microwave-assisted extraction) had a larger zone of inhibition for E. coli. Concluded, different methods of extraction change characteristics of the plant extract.

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How does the concentration of the extract affect its quality of effectiveness?

Previous findings describe antimicrobial activity directly related to the increase of the extract’s concentration. Tested in zones of inhibitions, the zone of inhibition linearly increases by the microgram (μg/ml) (larger zone of inhibition = better at combating bacteria). Phenolic compounds are used in cleaning materials, extracts richer in such content possess more antimicrobial activities in many organisms. Some bacteria/viruses will be more sensitive to extracts than others.

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Which extracts are associated with each viral disease?

There are many medicinal plants that treat oral infections or are consumed. Common herbs go as follows:

• Chamomile flowers can be used to induce rest and relaxation, treat rashes, wounds, or as a dose to ease vomiting during chemotherapy.
• Echinacea leaves, stalks, or roots are often used to treat colds, illnesses, coughs, flu, infections, and wounds-in-healing. Mostly used for respiratory infections and illnesses. Some may be allergic to this plant, since they are part of the daisy family.
• Feverfew leaves, as the name suggests, are used to treat fevers. They can prevent migraines and treat arthritis. Though, chewing feverfew leaves can lead to oral infections like ulcers, or upset your digestive system, and should not be used with blood thinning drugs.
• Garlic cloves are proven to fight germs, protect your heart, and reduce body inflammation. They may also improve cholesterol levels and blood pressure. Results may vary, they may induce bleeding and should not be taken with other blood thinning medicines.
• Ginger is used to treat motion sickness and nausea. Can be helpful during pregnancy or chemotherapy, and is currently being investigated on whether it has anticancer properties in surgery. Has strong antioxidant effects, but can induce heartburn, gas, and bloating.
• Ginseng is a Korean herb that isn’t certified with health benefits. Ginseng is said to boost energy, balance the body, and hormonally stimulate sex drive. Approved by FDA and is deemed safe to consume, but should not be taken with other medicines and should not be consumed by diabetic people.
• Gingko is a Chinese herb that is said to treat asthma, bronchitis (throat), tinnitus (auditory ringing or other sounds heard without an external source), and fatigue. Stated to improve energy, the real symptoms and side effects of ginkgo are unsure. Only leave extracts should be used, since gingko toxins in large amounts can cause seizure and even death. May increase bleeding. Do not use gingko with blood thinning medicines.

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How do properties in vaccines affect replication of viruses/bacterias?

Vaccines are designed to imitate a dose of the virus without its full infection. Vaccines train your immune system to build up antibodies to combat/fight off the bacteria. Through the lengthy process of developing immunity, side effects may result like nausea, dizziness, tiredness, etc. Building up resistance to the virus prevents replication in bacteria and viruses.

Vaccines, to have minute effects of the infection, so they include:

• Antigens: Trigger your immune system’s response. Often killed or weakened bacteria.
• Preservatives: Prevent harmful germs and microbes from growing within the substance. Only in miniscule or small amounts.
• Adjuvants: Boost/heighten your body’s immune system response. Examples of adjuvants are aluminum salts (present in some foods and breast milk) and squalene (present in some foods and is often a colourless oil).
• Additives: Gluten and/or organic substances used to keep vaccines effective while being stored.
• Substances from the manufacturing process: Formaldehyde, egg and yeast proteins, or antibiotics comprise a small fraction of the vaccine. Formaldehyde is naturally produced within the body, and is used to weaken or kill bacteria and viruses included in the vaccine.

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How do environmental factors influence the plant extract’s effectiveness on bacterial/viral models?

Environmental factors can put stress on plants, causing them to produce less material in flavonoids, phenolic acids, and things used for essential oils. Reduced production of these components can decrease their effectiveness in their medicinal functions. Previous studies done to test this prove that things like temperature and humidity and drought conditions directly impact quantities in these plant characteristics.

 

Once a plant is turned into traditional medicine, its qualities are impacted by its growth in its environment. Climate change and other environmental factors can alter and create variations of a plant’s medicinal and therapeutic qualities, which may not be suited for its traditional uses.

 

This is a concern for traditional medicine, since some people rely only on this form of clinical medication. Plants with changes in their medicinal traits may no longer have its traditional purpose. Plants may also go extinct or become scarce, leading to less production of these medicinal herbs. Following that, undiscovered healing capabilities in new species that cannot be tested will be forever unknown. Global health will be greatly impacted, as well as pharmaceutical businesses.

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Variables

Independent Variable (what you change)

• Type of plant extracts used (e.g.,  garlic, ginger, rosemary)
• Extraction method (aqueous vs. ethanol based)

Dependent Variable (what changes in response to the independent variable)

• Effectiveness of the plant extract against the virus/bacteria
• Zone of inhibition
• Reduction in viral/bacterial growth

Controlled Variable (what doesn’t change)

• Always use the same amount of plant material per mL of solvent
• Use the same volume of extract for each test plate
• All the cultures are incubated at the same temperature for the same time
• The same type of viral/bacterial model for all the tests
• Use the same growth medium for every agar plate

Control Group (for comparison)

• 1 agar plate with bacterial growth and no extracts to show what an untreated dish looks like
• 2 blank agar plates (compare to each other for 0 contamination)

 

1. Set up 6 nutrient agar plates using the powder, boiling water, and a refridgerator to cool down the dishes.
2. Collect and swab samples with a cotton swab from surfaces that are likely to bacterium and viruses. Introduce them to the petri dishes.
3. Incubate dishes, wait 4-6 days for bacteria to grow.
4. Set up a pot with water and place a container inside. Place the lid upside down on top of the pot, and turn on the stove. The condensation from the boiling water will collect on the top of the pan and drip into the container. This is our distilled water.
5. While waiting for enough water to collect start grinding ginger, rosemary, and garlic samples for ethanol extraction. Add the paste to the ethanol solvent with a 1:5 ratio.
6. Grind the same plants into paste, add them to the distilled water solvent with a 1:5 ratio.
7. Store extracts in vials, let them sit until agar plates have cultured bacteria.
8. Once agar plates are ready, photograph and label them with the matching extract and solvent that will be applied onto the petri dish lid.
9. Dig small wells/holes with a diameter of 5mm into the agar plates where the extracts will fill. Then, use a micropipette to fill the well with about 2g of plant extract
10. Incubate the plates again, document changes with photos and measure any visible zones of inhibition after the extract has been applied. Record in 12hr blocks.
11. Once growth has stopped, calculate the final inhibition zone in millimeter units. Compare and analyze the effectiveness of each plant extract and the impact of the different solvents.
12. Convert data into graphs with realistic expectations on how long medicinal effects last within the body (4-5 days). Complete project with results, observations, analysis, and conclusions.

DATA COLLECTED - 1d AFTER COLLECTING SAMPLES

OBSERVATIONS

• Agar plates are cloudy compared to untouched agar plates
• Small specks have grown in the agar

• Black fungus has grown around the edges of the plate
• Odd red streak was in one of the samples
• Some agar plates have less growth than others

 

DATA COLLECTED - 2d AFTER COLLECTING SAMPLES

OBSERVATIONS

• Clear outlines of swabbing bacteria were found inside the agar
• Fungus looks relatively the same
• Plates now have condensation on lids

 

DATA COLLECTED - 3d AFTER COLLECTING SAMPLES

OBSERVATIONS

• Bacterial cultures have grown a lot
• Specs are very prominent
• Fungus has grown larger
• Orange and yellow bacteria
• One sample distinctly has less bacteria than the other samples
• Large streaky bacteria found in one of the agar plates

 

DATA COLLECTED - 4d AFTER COLLECTING SAMPLES

OBSERVATIONS

• Relatively looks the same as yesterday
• Bacterial specks are a little more prominent

DATA COLLECTED - 12 hrs AFTER APPLYING EXTRACT WELLS

Gym Floor - Garlic (GcA or GcE)

1. Distilled : The zone of inhibition is 2mm in diameter
2. Ethanol : The zone of inhibition is 4mm in diameter

Sink Tap - Ginger (GrA or GrE)

1. Distilled : The zone of inhibition is 3mm in diameter
2. Ethanol : The zone of inhibition is 5mm in diameter

Beams - Rosemary (RmA or RmE)

1. Distilled : The zone of inhibition is 2mm in diameter
2. Ethanol : The zone of inhibition 3mm in diameter

 

OBSERVATIONS

• Extract inside the well is gone
• Cloudiness around the wells
• GrE has the most cloudiness surrounding the well
• Rm samples have the least amount of cloudiness around the well
• GcA has more cloudiness than GcE

 

DATA COLLECTED - 24hrs AFTER APPLYING EXTRACT WELLS

Gym Floor - Garlic (GcA or GcE)

1. Distilled : The zone of inhibition is 4mm in diameter
2. Ethanol : The zone of inhibition is 6mm in diameter

Sink Tap - Ginger (GrA or GrE)

1. Distilled : The zone of inhibition is 5mm in diameter
2. Ethanol : The zone of inhibition is 7mm in diameter

Beams - Rosemary (RmA or RmE)

1. Distilled : The zone of inhibition is 4mm in diameter
2. Ethanol : The zone of inhibition is 5mm in diameter

 

OBSERVATIONS

• Cloudiness has grown (not clear)
• Zone of inhibition is growing
• Red bacterial specks are more prominent
• More fungal growth on GrA

 

DATA COLLECTED - 36 hrs AFTER APPLYING EXTRACT WELLS

Gym Floor - Garlic (GcA or GcE)

1. Distilled : The zone of inhibition is 7mm in diameter
2. Ethanol : The zone of inhibition is 7mm in diameter

Sink Tap - Ginger (GrA or GrE)

1. Distilled : The zone of inhibition is 6mm in diameter
2. Ethanol : The zone of inhibition is 11mm in diameter

Beams - Rosemary (RmA or RmE)

1. Distilled : The zone of inhibition is 6mm in diameter
2. Ethanol : The zone of inhibition 9mm in diameter

 

OBSERVATIONS

• Cloudiness has grown (not clear)
• Zone of inhibition still growing
• Ethanolic extracts seem to be working well
• Rosemary samples had a growth spurt
• Fungus grew and spread around the petri dish
• Bottom of rosemary wells have a darker tint

 

DATA COLLECTED - 48hrs AFTER APPLYING EXTRACT WELLS

Gym Floor - Garlic (GcA or GcE)

1. Distilled : The zone of inhibition is 11mm in diameter
2. Ethanol : The zone of inhibition is 13mm in diameter

Sink Tap - Ginger (GrA or GrE)

1. Distilled : The zone of inhibition is 9mm in diameter
2. Ethanol : The zone of inhibition is 14mm in diameter

Beams - Rosemary (RmA or RmE)

1. Distilled : The zone of inhibition is 8mm in diameter
2. Ethanol : The zone of inhibition  is 10mm in diameter

 

OBSERVATIONS

• Cloudiness has grown (not clear)
• Large growth spurt in garlic extracts
• Zone of inhibition still growing
• Subtle changes

 

DATA COLLECTED - 60hrs AFTER APPLYING EXTRACT WELLS

Gym Floor - Garlic (GcA or GcE)

1. Distilled : The zone of inhibition is 13mm in diameter
2. Ethanol : The zone of inhibition is 13mm in diameter

Sink Tap - Ginger (GrA or GrE)

1. Distilled : The zone of inhibition is 11mm in diameter
2. Ethanol : The zone of inhibition is 15mm in diameter

Beams - Rosemary (RmA or RmE)

1. Distilled : The zone of inhibition is 9mm in diameter
2. Ethanol : The zone of inhibition  is 11mm in diameter

 

OBSERVATIONS

• Stunted in growth/slowed down
• Zone of inhibition appears the same as 12 hrs ago
• Subtle changes
• More condensation
• Bottom of rosemary wells look burnt

 

DATA COLLECTED - 72hrs AFTER APPLYING EXTRACT WELLS

Gym Floor - Garlic (GcA or GcE)

1. Distilled : The zone of inhibition is 13mm in diameter
2. Ethanol : The zone of inhibition is 15 mm in diameter

Sink Tap - Ginger (GrA or GrE)

1. Distilled : The zone of inhibition is 11mm in diameter
2. Ethanol : The zone of inhibition is 13mm in diameter

Beams - Rosemary (RmA or RmE)

1. Distilled : The zone of inhibition is 11mm in diameter
2. Ethanol : The zone of inhibition  is 14mm in diameter

 

OBSERVATIONS

• Cloudiness looks the same
• Condensation is more built up
• GrE fungus grew
• GrE inhibition zone decreased
• Some values remain the same

This experiment investigated the antibacterial effects of garlic, ginger, and rosemary extracts against bacteria collected from surfaces. The extracts were applied using both distilled water and ethanol solvents to determine their effectiveness. The zones of inhibition were measured overtime to analyze the antibacterial properties of each extract and compare their efficacy in different solvents. The inhibition zones formed around the extract wells indicated the antiviral activity of each plant extract. Our measurements showcased that ethanolic extracts generally had larger zones of inhibition compared to aqueous extracts, suggesting the ethanol was the more effective solvent in extracting antibacterial compounds. For garlic, the ethanol extract produced a final inhibition zone of 15mm, whereas the water-based extract reached only 13mm. This trend was consistent for ginger where the ethanol extract reached 13mm compared to 11mm for the water-based extract. Similarly, rosemary's ethanol extract produced a 14mm zone, with the water-based extract reaching 11mm. These results indicate that ethanol extracted a higher concentration of bioactive compounds responsible for antiviral activity.

The progression of inhibition zones over time showed that bacterial suppression became more pronounced after 24 hours with significant increases observed by 48 hours. For instance, garlic extract in the ethanol expanded from 4mm at 12 hours to 13mm at 48 hours, while ginger and rosemary followed similar patterns. This demonstrates that antiviral compounds need time to diffuse and take proper effect.

Throughout the experiment, several qualitative changes were noted. Initially, bacterial cultures appeared as specks and streaks growing denser over time. The presence of fungus, condensation, and red bacterial specs suggested microbial diversity in the collected samples. The cloudiness around extract wells also indicated bacterial inhibition, but it also implied possible interactions between the extract in the medium.

Garlic showed the highest antiviral activity followed by rosemary while ginger was the least effective in the end. There is evidence of antimicrobial resistance within the ginger inhibition zone tests, referring to the size decrease in diameter between 60 hrs and 72 hrs. This is likely due to the Allicin in the garlic; a compound that blocks viral entry and replication of microorganisms.

The result confirms that plant extracts can inhibit viral growth. However, their effectiveness varies based on type, solvent, the time of incubation, and the bacteria itself.

Despite following a controlled methodology, some errors or inconsistencies may have influenced our results. One potential issue was contamination, as fungal growth was observed on the agar plates. There is a possibility that contamination may have interfered with bacterial growth and inhibition zone measurements. Additionally, variations in the thickness of the agar, the depth of extract wells, or uneven spreading of bacteria could have caused slight discrepancies in inhibition zone sizes. Another influencing factor is the absorption of the extracts into the nutrient agar. Our observations indicated that the extracts were absorbed very quickly, possibly altering the concentration and the distribution. Environmental conditions, such as temperature fluctuations and condensation in the petri dishes, may have also impacted the bacterial growth rates. Lastly, human measurement errors when assessing the inhibition zones could also have introduced minor inconsistencies in our data collection.

This experiment demonstrated that garlic, ginger, and rosemary extracts have antiviral properties, with garlic being the most effective. Ethanol extracts consistently showed greater antibacterial activity than aqueous extracts, emphasizing the importance of solvent choice in these studies. Our findings suggest the potential applications for plant-based antimicrobials do in fact show high promise. Further studies could explore the mechanisms behind these effects and test a broader range of bacteria for comprehensive results and changes in global health. Addressing potential errors and refining methods, such as ensuring consistent sample preparation and minimizing external contaminants, would improve the accuracy and reliability of future experiments.

 

 

We were correct with our hypothesis. The tested extracts did result in zones of inhibition, which correlates to its antiviral and antibacterial properties. Using the agar well diffusion method, the data states that over the course of 72hrs, the garlic extracts were the most effective with the largest zones of inhibition, followed by rosemary and ginger. Furthermore, the ethyl alchohol solvent extracted more antimicrobial compounds than the distilled water solvent. With further research and experiments, we can investigate the potential of plant extracts and their clinical effects, leading to a beneficial outcome that can contribute to global health and safety.

Antiviral plant extracts could be used in real life to help treat and prevent viral infections in humans and animals alike. Multiple plant compounds have the ability to interfere with the way viruses infect cells or replicate inside the body. For example, extracts from elderberries, licorice root, and green tea have been studied for their ability to reduce the severity of infections like the flu, common cold, and even some respiratory viruses. These plant based treatments can be used in herbal medicines, teas, and even supplements to boost the immune system and help fight off viral illnesses. This is why it’s common for people to take echinacea tablets when sick, as it can easily help ward off and reduce symptoms of common cold and flu. 

Some plant extracts also show promise in treating more serious viral infections. Certain compounds from medicinal plants like Andrographis and echinacea have been dengue, fever, herpes and even HIV. These natural extracts may not be able to completely cure the infections, but they could help reduce symptoms and also relieve pain. In areas with limited access to conventional antiviral drugs, plant-based treatments could provide a more affordable and accessible option for managing viral illnesses. 

In the future, antiviral plant extracts could probably be administered in various ways, depending on the specific properties of plant compounds. The most common and easy method would be oral administration, where the extracts are taken as capsules, teas or syrups. Green tea and echinacea are taken to support the immune system. Some compounds like those from licorice root, can be formulated into lozenges or cough syrups to provide direct and quick relief for throat and respiratory infections. Topical application would be another effective method especially for skin related infections. Ointments, creams, and essential oil blends derived from plants like tea tree, neem or aloe vera can be applied directly to the skin to help treat sores, warts and other skin conditions. For more serious infections, plant based antivirals could be administered through an IV once more research is done in this field. As of right now this method is safer for synthetic drugs, however ongoing research is exploring more ways to extract and purify plant extracts to be safely injected for faster, targeted effects. Additionally, some plant extracts may be able to be integrated into vaccines and used alongside other treatments to enhance their effectiveness. 

Overall, plant extracts as antivirals have a bright future. With more research and development in this field, they may actually be able to provide a ton of possibilities for preventing and managing viral infections naturally. 

 

In our plant extract experiment, we found that several slight sources of error affected our results. One of our major sources of error was the inconsistency in the concentration of the plant extracts. Since plant compounds can vary based on factors like how they were prepared, how long they were stored, and even the exact method used for extraction, slight variations may have impacted the antiviral effectiveness observed. This most likely caused them to have stronger or weaker results than expected. Another likely source of error could have been contamination or interference. This is most likely due to our equipment, the distilled water, or the containers used. The conditions of the experiment, such as temperature, pH, or exposure time may have tampered with the results as well. If a plant extract is exposed to light, air , or heat for too long, some of the active compounds would have the chance to degrade, automatically decreasing the effectiveness. Similarly, if different samples were tested for slightly different durations or, possibly causing inconsistent results. Lastly measurement errors may have affected our findings. If I were to redo this experiment I would try to have more precise measurements, clearer controls, and also make sure to do many trials to ensure consistency throughout the experiment. 

 

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We would like to thank our parents for helping us gather all the materials needed for this project. We would also like to thank Ms. Rheinstein and Mr. Lahoda for coordinating and guiding us.