The Study of Some Therapeutic Effects of

Natural Medicines.

Presented By:

Wi’aam Abid JwairBachelor Degree in General Biology/College of Sciences

/University of Tikrit.

Supervised by:Assist. Prof.

Dr. Ausama Nadhum Najris.

جامعة تكريتكلية العلوم

قسم علوم الحياة

An Introduction:

Today the control of microbial action and redox status within the human body through

food and food components has become the most important field of research .

An imbalance between reactive oxygen species (ROS) and reactive nitrogen species

(RNS) formation and the capacity of antioxidant defenses has resulted in the occurrence

of oxidative stress. This imbalance instigate chronic diseases such as neurodegenerative

diseases related to ageing, as well as cancer and cardiovascular diseases by damaging

DNA, proteins and lipids.

New resistance mechanisms of microorganisms against antimicrobials are emerging and

spreading globally, which is caused by the misuse and overuse of antibiotics, which

encourages bacteria to develop new ways to overcome their effects. In addition,

antibiotics might possess an adverse effects on the body, which include hypersensitivity,

immunosuppressant and allergic reactions. Thereby, threatening our ability to treat

common infectious diseases, resulting in prolonged illness, disability, and death.

These situations forced scientists to explore for new anti-microbial and therapeutic

antioxidant efficacious substances from various sources, such as medicinal plants.

The medicinal plants that were used in this study were: Chamomile (Chamaemelum

nobile), licorice (Glycyrrhiza glabra) and mistletoe (Viscum album) which have been

used throughout the world as drugs and remedies for various diseases since ancient

times. Nowadays new researches have found that these medicinal plants possess many

therapeutic effects starting from antimicrobial, anti-inflammatory, and

immunostimulatory effects to antioxidant, hypotensive, vasodilator, cardiac

depressive, sedative, antispasmodic, and anticancer effects.

These therapeutic properties are because of the complex phytochemical content of

these plants. Some of these phytochemicals were detected through this research :

phenols, flavonoids, glycosides, triterpenes, alkaloids, carbohydrates,

phytosterols and Saponins.

It is believed that combinations between drugs and herbs can help to :

Increase antimicrobial activity.

Reduce toxic effects of medicinal agents.

Treatment of mixed infections and other infections.

Prevent need for long term of antibiotic use.

Prevent emergence of multidrug resistance bacteria by increasing the killing rate,

potentiating of drugs, prevention of drug elimination, a better effect in vivo.

Prevent resistant mutant population from emerging as the dominant strains since any

drug of the combine therapy could be effective against them.

The combination of two drugs can be;

Synergistic; in which two different compounds are combined to enhance their

individual activity.

Antagonistic: If the combination results in worsening effect.

Additive: which is less than synergistic but not antagonistic.

The first example for combinations was the synergistic activity of amoxicillin with

clavulanic acid have shown successful results against β-lactamase-producing strains of

Haemophilus ducreyi.

The aim of this study is to promote the use of herbal medicines; and to find alternative

ones for the synthetic drugs by means of exploring the efficacy of some therapeutic

effects of the studied plants, especially the antimicrobial effects against the multidrug

resistant microorganisms.

The objectives of this study:

1. Make more than one type of plant extracts namely, alcoholic extract and aqueous

extracts and

2. Find the differences in the detecting results for active compounds and relate their

presence to a certain effect in further lines of the study.

3. Explore the efficacy of the antimicrobial effect of the herbal extracts.

4. Find a way to increase the efficacy of standard antimicrobials against the resistant

microorganisms by combining the extracts with the standard antimicrobial in certain

ratios.

5. Explore the antimicrobial effect of the plant extracts by combining them.

Materials And Methods

Plants (Chamaemelum nobile, Glycyrrhiza glabra, Viscum album) were collected from Iraqi herbariums and were

identified by Dr. Shakir Mahdi and then they were extracted with 3 solvents. Methanol was used in estimating “Free

Radical Scavenging Activity”; while ethanol and water were used for “ The antimicrobial Potency Tests”.

Some Phytochemicals were detected: Phenols, alkaloids, Triterpenes, Flavonoids, Carbohydrates, Glycosides,

Phytosterols and Saponins.

The “ Total Phenolic Content” and “ Total Alkaloids” were estimated quantitatively.

The Standard Antibiotics that were used in “The Antibiotic Potency Test” were: Neomycin Sulfate, Erythromycin ethyl

succinate, Gramicidin, Gentamicin Sulfate, Tetracycline, Amoxicillin against each of : E. coli, Staphylococcus aureus,

Pseudomonas aeruginosa, Bacillus pumilus. While the Antifungals that were used were: Nystatin, Metronidazole,

Clotrimazole against Candida albicans and Saccharomyces cerevisiae.

The Microorganisms species mentioned above were confirmed by means of Vitek2 and methods mentioned in Oxoid

Manual (Bridson, 2006) (Only E. coli).

The antimicrobial potency of the ethanolic and aqueous plant extract were tested against: E. coli, Staphylococcus

aureus, Bacillus pumilus and Candida albicans. And the Antimicrobial Potency of “ Combining between the

antimicrobials with the plant extracts” were only tested against: E. coli and Candida albicans.

The combination that were made were : (25%+ 75%), (50%+50%),(75%+25%) of each of Plants Extracts and some

Antimicrobials (Gramicidin, Erythromycin and Nystatin). And this test was carried according to the lack of activity the

antibacterials and the plants extracts against E. coli.

Some plants extracts were combined together in a 50:50 percentages to test the antimicrobial effects of their combining.

Extracting

Ethanolic and Aqueous Extracts

Phytochemical Screenings

Estimation of Total Phenolic Content

Estimation of Total Alkaloid

ContentAntimicrobial Tests

Standard Antimicrobial

Potency

Antimicrobial Potency of The Plant Extract

Plant + Antimicrobials Antimicrobial Potency

50% Plant Extracts+50% Antimicrobials

25% Plant Extracts+ 75% Antimicrobials

75% Plant Extract+ 25%

Antimicrobials

Plants Mixtures Antimicrobial

Potency

50% Gg+50% Cn

50%Va+50%Cn

50%Va+50%Gg

Methanolic Extracts

Free Radical Scavenging Potency

22.3

00

13.5

16.5

11.8

Inh

ibit

ion

Zo

ne

s (m

m).

Treatments (mg/ml).

The Antibacterial effects of Combining(Gramicidin+ Alcoholic Viscum album Extract)

Against E. coli.

Neomycin Gramicidin Viscum album

75%Va.+25%Gr. 50%Va.+50%Gr. 25%Va+75%Gr.

22.3

00

16.2

13

10

Inh

ibit

ion

Zo

ne

s (m

m).

Treatments (mg/ml).

The Antibacterial Effects of Combining (Erythromycin + Alcoholic Viscum album Extract)

Against E. coli.

Neomycin Erythromycin Viscum album

75%Va Extract+25%Ery. 50%Va Extract+50% Ery 25%Va Extract+75%Ery

22.3

00

1413

12

Inh

ibit

ion

Zo

ne

s (m

m).

Treatments (mg/ml).

The Antibacterial Effects of Combining (Gramicidin + Aqueous Viscum album Extract)

against E.coli.

Neomycin Gramicidin Aq. Va Extract

75%Va+25%Gr. 50%Va+50%Gr. 25%Va+75%Gr.

22.3

00

1412

10.5

Inh

ibit

ion

Zo

ne

s (m

m).

Treatments (mg/ml).

The Antibacterial Effects of Combining (Erythromycin + Aqueous Viscum album Extract)

Against E.coli.

Neomycin Erythromycin Viscum album

75%Va+25%Ery. 50%Va+50%Ery. 25%Va+75%Ery.

22.3

00

1415

12

Inh

ibit

ion

Zo

ne

s (m

m).

Treatments (mg/ml).

The Antibacterial Effects of Combining (Gramicidin + Alcoholic Chamemelum noble Extract

Against E.coli.

Neomycin Gramicidin Chamaemelum noble

75%Cn+25%Gr. 50%Cn+50%Gr. 25%Cn+75%Gr.

22.3

00

17.1

1310.8

Inh

ibit

ion

Zo

ne

s (m

m).

Treatment (mg/ml).

Erythromycin + Alcoholic Chamaemelum noble Extract.

Neomycin Erythromycin Chamaemelum noble

75%Cn+25%Ery. 50%Cn+50%Ery. 25%Cn+75%Ery.

22.3

00

1413.811.8

Inh

ibit

ion

Zo

ne

s (m

m).

Treatments (mg/ml).

The Antibacterial Effects of Combining (Gramicidin + Aqueous Chamaemelum noble Extract)

Against E.coli.

Neomycin Gramicidn Chamaemelum noble

75%Cn+25%Gr. 50%Cn+50%Gr. 25%Cn+75%Gr.

22.3

00

16.113.8

10.5

Inh

ibit

ion

Zo

ne

s (m

m).

Treatments (mg/ml).

The Antibacterial Effects of Combing(Erythromycin + Aqueous Chamaemelum noble Extract)

against E.coli.

Neomycin Erythromycin Chamaemelum noble

75%Cn+25%Ery. 50%Cn+50%Ery. 25%Cn+75%Ery.

22.3

00

18.217.315

Inh

ibit

ion

Zo

ne

s (m

m).

Treatments (mg/ml)

The Antibacterial Effects of Combining (Gramicidin + Alcoholic Glycyrrhiza glabra Extract)

Against E.coli.

Neomycin Gramicidin Glycyrrhiza glabra

75%Gg+25%Gr. 50%Gg+50%Gr. 25%Gg+75%Gr.

22.3

00

18.216.1

11

Inh

ibit

ion

Zo

ne

s (m

m).

Treatments (mg/ml).

The Antibacterial Effects of Combining (Erythromycin + Alcoholic Glycyrrhiza glabra Extract)

Against E.coli.

Neomycin Erythromycin Glycyrrhiza glabra

75%Gg+25%Ery. 50%Gg+50%Ery. 25%Gg+75%Ery.

22.3

0

0

1412.5

11.2

Inh

ibio

tio

n Z

on

es

(mm

).

Treatments (mg/ml).

The Antibacterial Effects of Combining(Gramicidin + Aqueous Glycyrhiza glabraExtract)

against E.coli.

Neomycin Gramicidin Glycyrrhiza glabra

75%Gg+25%Gr. 50%Gg+50%Gr. 25%Gg+75%Gr.

22.3

00

19.217.1

11.2

Ihib

ibti

on

Zo

nez

(m

m).

Treatments (mg/ml).

The Antibacterial Effects of Combing(Erythromycin + Aqueous Glycyrrhiza glabra Extract)

against E.coli.

Neomycin Erythromycin Glycyrrhiza glabra

75%Gg+25%Ery. 50%Gg+50%Ery. 25%Gg+75%Ery.

22.3

000000

11.1

Inh

ibit

ion

Zo

ne

s (m

m).

Treatments (mg/ml).

The Antibacterial Effects of Combining(Chamaemelum noble and Viscum album)

against E. coli.

Neomycin 1000mg/ml Alc. Va500mg/ml Alc. Va 250mg/ml Alc. Va1000mg/ml Alc. Va 500mg/ml Alc. Cn250mg/ml Alc. Va (250+250)mg/ml Alc Va+Cn

13.8

0

10.3

0

10

14

Inh

ibit

ion

Zo

ne

s (m

m).

Treatments(mg/ml).

Antimicrobial Effects of Combining(Alcoholic Extracts of Glycyrrhiza glabra And Viscum album)

Against Bacillus pumilus .

Neomycin 250mg/ml Alc. Gg

500mg/ml Alc. Gg 250mg/ml Alc Va

500 mg/ml Alc. Va (250+250)mg/ml Alc. Gg+Va

15

0

19.2

0

20

0

19

Inh

ibit

ion

Zo

ne

s (m

m)

Treatments (mg/ml).

The Antifungal Effects of Combining(Alcoholic Plants Extracts with Nystatin)

against Candida albicans.

Nystatin 250mg/ml Alc. Cn 25%Cn+75% Nys.

250mg/ml Alc. Gg 25% Gg+75% Nys. 250% Alc. Va

25% Va+75% Nys.

15

0

11.21110

Inh

ibit

ion

Zo

ne

s (m

m).

Treatments (mg/ml)

The Antifungal Effects of combining(Chamaemelum nobile with Other Plant Extracts)

Against Candida albicans.

Nystatin 250mg/ml Alc. Cn

500% mg/ml Alc. Cn 250+250 mg/ml Cn+Gg

250+250 mg/ml Cn+Va

15

00

1110

Inh

ibit

ion

Zo

ne

s (m

m).

Treatments (mg/ml).

The Antifungal Effects of Combining(Glycyrrhiza glabra with other Plants Extracts)

against Candida albicans.

Nystatin 250 mg/ml Alc. Gg

500 mg/ml Alc. Gg 250+250 mg/ml Gg+Cn

250+250 mg/ml Gg+Va

15

0

8

1010

Inh

ibit

ion

Zo

ne

s (m

m).

Treatments (mg/ml).

The Antifungal Effects of Combining(Viscum album with Other Plants Extracts)

against Candida albicans.

Nystatin 250 mg/ml Alc. Va

500 mg/ml Alc. Va 250+250 mg/ml Va+Cn

250+250 mg/ml Va+Gg

Conclusions:

1. Plant extracts contained a very complex structure with the active ingredients present in the form of

natural organic compounds.

2. The process of extraction for a particular compound is dependent on the solubility of the component in

the solvent (water or organic solvent). The process and extraction system are constantly different with

every product and compound.

3. The crude extracts of the tested plants demonstrated good potential antibacterial activities. The

potential to develop antimicrobial compounds from higher plants appears rewarding as it will propel to

the expansion of phytomedicines to turn against multidrug resistant microbes.

4. Combinations, whether they were randomly done or by specific preparations and percentages, have an

effect on the activity of inactive antimicrobials.

5. The synergistic effect of the combinations promotes the use of whole plants as beverages or other way

of consuming. Because while each individual active compound has a variety of health benefits, it has to

be appropriate to assume that there is a good chance that the synergic interaction could lead into further

health benefits.

6. It seems that every natural product has a certain capacity of scavenging free radicals because of the

variety of active ingredients present in them.

Recommendations:

1. Using more solvents in plant extraction.

2. Screening for more active compounds in the plants and exploring their individual effects on many

researching aspects; and then making a comparison between the efficacy of each active compound and

another in each aspect of researching.

3. Making further studies on the physiological effects of the plant extracts.

4. Making more exploring in the synergism between plant extracts and antimicrobials to find good solutions

for antimicrobial resistance.

5. Consuming more natural food products, since they possess the capacity of scavenging free radicals, to

enhance immunity and the whole body health.

Thank You All for

Listening