Transcript Science Fair Project

Antimicrobial Essential
Oils
Scientific research behind thyme, oregano
and clove
Nicole Stevens
Scope
 Parasitic infection is worldwide
 Outranks cancer as #1 global killer
 The Center for Disease Control
estimates:
 62 million cases of infectious illness in
the US yearly.
 Common cold caused by any of 200
different viruses, of which 30%-50%
remain unidentified.
 10%-15% of adult colds are caused by
viruses also responsible for more
serious illnesses
Menagerie
 There are over 1,000 species of parasites that
can live in your body.
 Doctors can only identify and test for 5% of
these parasites.
Pneumonia
Image source: http://www.hsph.harvard.edu/mizgerdlab/web_site/images_on_background/sem.gif
 Several parasites have emerged as significant
causes of food borne and waterborne diseases.
 30% of parasites live in our digestive system;
 70% live throughout the body including the
blood, brain, eyes and sinus.
Asymptomatic
 Not all parasites produce acute illness—
many cause subtler chronic symptoms










Flu-like symptoms
Allergies
Weight loss
Ravenous appetite
Abdominal bloating, gas, cramping,
diarrhea, nausea
Fever
Joint pain
Disturbed sleep
Muscle cramps
Sugar cravings
Image Source:http://health.utah.gov headache.jpg
Costs
 In the US annually:





$3.5 billion spent on OTC cough remedies
$2 million spent on “natural” medicines that fight colds
$1.3 billion spent on direct medical costs to fight flus
70 million work-loss days associated with illness
Lost productivity in the workplace costs businesses $15 billion
Image source: http://www.bainbridgepharmacy.com/pharmacy/
Tiny Creatures, BIG Problem
“The worldwide overuse of antibiotics has caused microorganisms to develop resistance to
current antibiotics and to become virulent.”
(Compendium of Continuing Education in Dentistry, Jan. 9, 1988, Harry G. Meeker, DMD)
 National Institutes of Health:
 About 90,000 deaths annually from infections that don’t respond to treatment
 More and more bacteria (such as Staph. aureus and numerous gut bacteria) are
resistant to the most powerful antibiotics available
 Centers for Disease Control:
 Doctors prescribe antibiotics to children 65% of the time if they feel parents expect
them, even if not needed
 Nearly all significant bacterial infections in the world are resistant to the most
commonly prescribed antibiotics
Our Solution
“Microorganisms, however, do not appear to develop tolerance or resistance to the
antibacterial effects of essential oils such as clove oil (eugenol) and thyme oil (thymol).
Such oils provide an effective, powerful, and cost-effective means of infection
control…”
(Compendium of Continuing Education in Dentistry, Jan. 9, 1988, Harry G. Meeker, DMD)
 Essential Oils: antimicrobial champions!
 Thyme (>55% thymol)
 Clove (>80% eugenol)
 Oregano (>60% carvacrol)
Safety
 Eugenol (from clove EO), thymol (from thyme EO), and carvacrol (from
oregano EO) have been used in dentistry since the 19th century for root
canals, temporary fillings and cements, periodontal therapy, and abscesses
 Bernard Schechter, DDS
 Currently designated GRAS by the FDA
Thyme
THYME ESSENTIAL OIL ANTIMICROBIAL ACTIVITY
BACTERIA (gram negative/positive)
FUNGI/MOLDS
Acinetobacter calcoacetica10
Haemophilus influenzae5
Pseudonomas fluorescens20
Aspergillus flavus1,13,18,22
Actinobacillus actinnomycetemcomitans26
Klebsiella pneumoniae10
Saccharomyces cerevisiae20
Aspergillus fumigatus5,14
Actinomyces viscosus26
Lactobacillus curvatus24
Salmonella enterica2
Aspergillus ochraceus1
Aeromonas hydrophila10
Lactobacillus plantarum10
Salmonella pullorum10
Aspergillus parasiticus1,6,12,23
Alcaligenes faecalis10
Lactobacillus sake24
Sarcima spp.20
Candida albicans21
Bacillus cereus9
Leuconostoc cremoris10
Selenomonas artemidis19
Fusarium moniliforme1
Bacillus subtilis10,20
Listeria innocua27
Serratia liquefaciens24
Fusarium proliferatum12
Beneckea natriegens10
Listeria monocytogenes2,3,17
Serratia marcescens10,20
Fusarium solani14
Brevibacterium linens10
Micrococcus luteus10
Staphylococcus aureus5
Penicillium citrinum15
Brochothrix thermosphacta
10
Micrococcus spp
20
16
7
Salmonella sp.
Penicillium digitatum
Campylobacter jejuni2
Morazella sp.10
Streptococcus pneumoniae5
Penicillium expansum11,14
Carnobacterium piscicola24
Mycobacterium phlei20
Streptococcus pyogenes5
Phytophtora cinnamomi21
10
4
Streptococcus milleri
25
Pyrenochaeta lycopersici
21
Citrobacter freundii
Photobacterium phosphoreum
Clostridium sporogenes10
Peptostreptococcus anaerobius25
Streptococcus mitis25
Rhizopus oryzae14
Eikenella corrodens26
Porphyromonas gingivalis1,2,19
Streptococcus mutans25
Thrichophyton mentagrohphytes21
Enterobacter aerogenes10
Prevotella buccae25
Streptococcus sanguis25
Verticillium dahliae21
Enterococcus faecalis10
Prevotella intermedia25
Streptococcus sobrinus19
Erwinia carotovora10
Escherichia coli 0157:H72
Prevotella nigrescens26
Prevotella oris25
Treponema vincentii26
Yersinia enterocolitica10
Escherichia coli10,20
Flavobacterium suaveolens10
Proteus vulgaris10
Pseudonomas aeruginosa10
PROTOZOA PARASITES
Leishmania major8
Trypanosoma brucei8
1 Soliman and Badeaa, 2002, 2 Friedman et al., 2002, Bart et al. 2004, 3 Smith-Palmer et al., 2002, 4 Mejlholm and Dalgaard, 2002, 5 Inouye et al. 2001, 2000, 6 Tantaoui-Elaraki and Beraoud, 1994, 7 Daferera et al., 2000, 8 Mikus et
al., 2000, 9 Ultee et al., 2000, 10 Dorman and Deans, 2000, 11 Venturini et al., 2002, 12 Juglal et al., 2002, 13 Lopez-Malo et al., 2002, 14 Inouye et al., 1998, 15 Vazquez at al., 2001, 16 Weissinger et al., 2001, 17 Ettayebi et al.,
2000, 18 Montes-Belmont and Carvajal, 1998, 19 Shapiro and Guggenheim, 1995, 20 Farag et al., 1987, 21 Giamperi et al., 2002, 22 Mahmoud, 1994, 23 Farag et al., 1989, 24 Ouattara et al., 1997, 25 Didry et al., 1995, 26 Shapiro et
al., 1994, 27. Olasupo NA, et al., 2004
Thyme vs. Microbes
 A 1995 study by Nicole Didry at the College of Pharmaceutical and
Biological Sciences in Lille, France
 Thyme oil at very small concentrations (<500 ppm) killed the pathogenic
organisms responsible for the tooth decay, gingivitis, and bad breath








Streptococcus mutans,
S. sanguis
S. milleri
S. mitis
Peptostreptococcus anaerobius
Prevotella buccae
P. oris
P. intermedia
Essential Oil vs. Oral Pathogens
KILL TIMES AGAINST ORAL PATHOGENS
Fusobacterium nucleatum
Streptococcus mutans
Prevotella intermedia
Lactobacillus casei
Candida albicans ATCC 28366
Candida albicans ATCC 18804
Essential Oil
<0.5 min
<0.5 min
<0.5 min
<0.5 min
<0.5 min
<0.5 min
TYPES of MOUTHRINSES
Stannous Fl Sterile Water Control
<0.5 min
>5 min
<1 min
>5 min
<2 min
>5 min
>2 min
>5 min
>5 min
>5 min
>5 min
>5 min
Pan et al., 1999
“Essential oils, such as thymol, menthol and eucalyptol, have been used in mouth rinses for over
100 years [and] have been documented to be antibacterial in laboratory tests. With the
association of microorganisms and plaque formation, and [their] suspected involvement with
carious lesions (tooth decay) and gingivitis, the effect of essential oils takes on new interest.”
Journal of the Society of Cosmetic Chemists
Thyme for Health
 Researchers at the Scottish Agricultural College
 Daily doses of thyme oil (42.5 mg/K of body
weight), drastically slowed age-related DHA
degradation in the brain
 Karesh Youdim and colleagues
 3.9 mg thyme and clove oils for 17 months
slowed DHA loss
Image source: http://www.nu-mega.com/images/Brain.jpg
 Jukic et al (2007)
 Thyme oil and derivatives slow destruction of
neurotransmitters
Thyme for Health
Protection of Brain Antioxidant Activity
thyme-treated
thymol-treated
control group
Youdim, Deans, 2000
Thyme: The Bottom Line
 Research shows us that thyme essential oil:
 Is potently antimicrobial
 Has been used for centuries in oral health (and
health in general!)
 May slow or reverse aging of brain tissue by
protecting DHA, antioxidants and
neurotransmitters
 May slow or prevent age-related eye tissue
damage by protecting DHA
Image source: http://www.dkimages.com/discover/previews/880/25092981.JPG
Clove
Clove: Long History
 Subjects awaiting an audience with Chinese emperors
were required to chew cloves to mask breath odors (207
B.C. to 220 A.D.)
 Avicenna treated “putrifaction” of the teeth and gums
with clove pills (~1000 A.D.)
 New England physician, Thomas Palmer, wrote in 1696:
Image source: http://www.aromaticsinternational.com/img/oilthumbs/oils_clb.jpg
 “Oyl of cloves—stayeth ye putrifaction of the bones in old and
new ulcers . . . It heals wounds, diseases, wind, digesteth cold
humours. Causes a sweet breath.”
 Thomas Berdmore, Operator for the Teeth to King
George III, treated toothache with “acrid aromatic
substances” including mace and cloves (late 1700s)
Clove: Long History
 The Island of Ternate in the Moluccas
Archipelago was the site of extensive clove
cultivation in the 16th century
 Dutch destroyed the clove trees to protect their
monopoly
 Native inhabitants were devastated by disease
(60,000+ thought to have died)
 Researchers attributed the sudden onset of epidemics
to the lack of “clove leaves and flowers sufficient to
purify the air.”
 Some East Indies natives still wear clove stuck to
nostrils and lips so demons do not enter there
Image source: http://www.altcancer.com/phyto/images/clove_tree.jpg
Safety
 Safety confirmed by NTP in lifetime animal studies (1983, Technical Report
No. 223, National Toxicology Program)
 Ames salmonella assay also showed eugenol to be antimutagenic (1995,
Azizan & Blevins, East Tennessee State University)
 Animals studies at the University of Wisconsin Medical School achieved similar results
in an animal model.
 The Joint FAO/WHO Expert Committee on Food Additives estimated an
acceptable human daily intake of eugenol of up to 2.5mg / kg body weight
 The German Commission E monograph prescribes mouthwashes consisting
of 1 to 5% clove essential oil as an oral antiseptic and topical anesthetic,
stating that it has “antibacterial, antifungal, antiviral” action
Clove vs Pathogens
BACTERIA (gram negative/positive)
5,8
FUNGI/MOLDS
5,8
Acinetobacter calcoacetica
Actinobacillus actinnomycetemcomitans2
2
Actinomyces viscosus
Aeromonas hydrophila5,8
5,8
Alcaligenes faecalis
Bacillus sp.18
Bacillus subtilis5,6,8,19,21
5,8
Beneckea natriegens
Brevibacterium linens5,8
Brochothrix thermosphacta5,8,9
Campylobacter jejuni10
9
Carnobacterium piscicola
Citrobacter freundii5,8
Clostridium sporogenes5,8
Eikenella corrodens2
Enterobacter sp. 18
Enterobacter aerogenes5,8
5,22
Enterococcus faecalis
Erwinia carotovora5,8
Escherichia coli5,6,8,19
10
Escherichia coli 0157:H7
5,8
Flavobacterium suaveolens
Klebsiella pneumoniae5,8
9
Lactobacillus curvatus
5,8
Lactobacillus plantarum
Lactobacillus sake9
Micrococcus luteus
Micrococcus spp6,12
5,8
Morazella sp.
Mycobacterium phlei6,19
2,3
Peptostreptococcus anaerobius
Photobacterium phosphoreum14
Porphyromonas gingivalis1,2
3
Prevotella buccae
Prevotella intermedia1,3
Prevotella nigrescens2
Prevotella oris3
5,8
Proteus vulgaris
Pseudonomas aeruginosa5,8,19
Pseudonomas fluorescens6,9
Saccharomyces cerevisiae6
Salmonella enterica10
Salmonella pullorum5,8
6
Sarcima spp.
Selenomonas artemidis2
Serratia liquefaciens9
5,6,8
Serratia marcescens
18
Staphylococcus sp.
Staphylococcus aureus5,6,8,19
3
Streptococcus milleri
3
Streptococcus mitis
Streptococcus mutans3
Leuconostoc cremoris5,8
Listeria monocytogenes10,13
Listeria innocua 21
Streptococcus sanguis2,3
Streptococcus sobrinus2
Treponema vincentii2
Yersinia enterocolitica5,8
VIRUSES
16
Aspergillus flavus
Aspergillus niger19
7,11
Aspergillus parasiticus
Candida albicans19
12
Colletotrichum musae
Fusarium moniliforme11
Fusarium proliferatum12
12
Lasiodiplodia theobromae
Mucor sp.19
Penicillium chrysogenum19
Penicillium citrinum 17
15
Penicillium expansum
Rhizopus sp.19
Clove vs. Microbes
Herpes virus I and II
4
1 Cai and Wu, 1996
2 Shapiro et al., 1994
3 Didry et al., 1994
4 Benencia et al., 2000
5 Dorman and Deans, 2000
6 Farag et al. 1989
7 Farag et al. 1989
8 Lis-Balchin et al., 1997
9 Ouattara et al., 1997
10 Friedman et al., 2002
11 Juglal et al., 2002
12 Ranasinghe et al., 2002
13 Smith-Palmer et al., 2002
14 Mejlholm and Dalgaard, 2002
15 Venturini et al., 2002
16 Lopez-Malo et al., 2002
17 Vazques et al., 2001
18 Moleyar and Narasimham, 1992
19 Yousef and Tawil, 1980
20 Chami F et al., 2004
21 Olasupo NA, et al., 2004.
22 Kayaoglu G et al., 2005
Clove vs. Microbes
 Eugenol strongly inhibits
growth of HSV I and
HSV II
 Eugenol significantly
delayed the onset of
herpetic lesion on the
eyes of infected animals
Clove vs. Microbes
 Bacteria and fungi tend cluster together in
clusters called biofilms that serve as a powerful
protective mechanism
 Biofilms make the microorganisms much more
difficult to kill
 Clove is one of the few natural compounds that
can disrupt these biofilms
 Recent studies have shown that eugenol disrupts
Candida biofilms.
[1] Ahmad et al., “Antimicrobial activity of clove oil and its potential in the treatment of vaginal
candidiasis,” 2005
[2] He et al., “In vitro activity of eugenol against Candida albicans biofilms.” 2007.
Image source: http://www.ansci.wisc.edu/facstaff/Faculty/pages/albrecht/albrecht_web/Programs/microscopy/images/andes%20biofilm%20sem.jpg
Clove for Health
 According to a test developed by USDA researchers at Tufts
University clove bud oil is the most powerful antioxidant
known
ORAC score per 100g
Clove Bud Essential Oil
Nutmeg Essential Oil
1,078,687
158,130
Prune
5, 700
Blueberry
2,400
Raspberry
1,200
Orange
750
Clove for Health
 Nagababu et al (1995) showed that
eugenol protected against CCl4
hepatotoxicity (liver damage)
 Due to eugenol’s amazing free
radical scavenging activity!
 Also showed that eugenol protected
against lipid damage from free
radicals (1994)
Image source: http://www.portfolio.mvm.ed.ac.uk/studentwebs/session3/7/Treatm2.jpg
Clove: The Bottom Line
 Research shows us that clove essential oil:




Is potently antimicrobial
Is an incredibly powerful antioxidant
Is an amazing anti-inflammatory
Has been used for centuries in oral health
(and health in general!)
 Shown to protect DHA in retinal tissue
Image source: http://www.womens-health-symmetry.com/images/CloveBudLeaf.gif
Oregano
Carvacrol
Oregano vs. Microbes
Oregano vs. Microbes
 A 2001 Georgetown University Study
found that oregano oil at a concentration of
0.25 mg/ml completely inhibited candida
growth
 Entire control group died in 10 days (100%
mortality)
 Mice infected with Candida fed 8.6 mg of
oregano oil for 30 days had only 20%
mortality
Survival of Candida-infected Subjects
100%
90%
100%
80%
80%
70%
60%
50%
40%
30%
20%
Manohar V, Ingram C, Gray J, Talpur NA, Echard BW,
Bagchi D, Preuss HG. Antifungal activities of origanum oil
against Candida albicans. Mol Cell Biochem. 2001
Dec;228(1-2):111-7.
0%
0%
10%
0%
10 days
Oregano-Fed
30 days
Control
Oregano vs. Microbes
 Adam et al (1998)
 Oregano essential oil at a 1/50,000
dilution caused 95% kill of human
pathogenic fungi
 In vitro and in vivo
 No mutagenic activity!
 Daouk et al (1995)
Image source: http://129.215.156.68/Images/Asexual%20structures%20of%20Aspergillus%20niger.jpg
 Oregano oil at 0.1μl / mL strongly
inhibited common (and deadly) food
fungi
Oregano vs. Microbes
 Research at Weber
State University by
Sue Chao found that
12 microliters of
oregano had the
antibacterial
equivalence of 10
units of penicillin.
Oregano EO
Penicillin (10 units)
microLiters
3
E COLI Zone of
Inhibition (cm)
2.4
0
STAPH AUREUS
Zone of Inhibition
microLiters
(cm)
Oregano EO
12
3.2
Penicillin (10 units)
3.2
Streptococcus pneumoniae before Oregano Oil
exposure
Streptococcus pneumoniae after Oregano
Oil exposure
Oregano vs. Microbes
 Harry Preuss, MD, at Georgetown found that
oregano oil (in particular, carvacrol) works as well
as antibiotics on Staphylococcus aureus.
 Oregano oil also killed E. coli, Heliobacter pylori,
Bacillus anthracis, and Klebsiella pneumoniae
 Penicillin resistant
 Phase 2: animal studies
 50% of mice fed oregano oil for 30 days survived
Staph. aureus infections
 Control mice died within 3 days
Image source: http://www.biology4kids.com/extras/dtop_micro/7821_580.jpg
Oregano for Health
 Carvacrol and thymol in oregano
oil are antioxidants
 Slamenova et al (2007)
 Very small concentrations of
carvacrol protected cells against
DNA damage from hydrogen
peroxide
 Carvacrol and thymol themselves
did not damage DNA
Image source: http://www.ch.cam.ac.uk/magnus/molecules/nucleic/dna1.jpg
Oregano: The Bottom Line
 Research shows us that oregano essential
oil:
 Is potently antimicrobial
 Many microbes that are resistant to common
antibiotics
 Is a powerful antioxidant that helps protect
DNA from damage
 Is not mutagenic or cytotoxic to cells
Image source: http://www.westcountryorganicherbs.co.uk/oregano.JPG
So What?
 Infectious disease and growing antibiotic
resistance are major threats to our health
 Essential oils, especially thyme, oregano and
clove, can help!
Image source: http://www.thevibecenter.com/images/LivingOils2.jpg





Powerful antimicrobials
No resistance demonstrated
Synergistic: also support health in other ways
Internal and external benefits
Centuries of safe and successful use