Molecular and Cellular Biochemistry 228: 111–117, 2001.

© 2001 Kluwer Academic Publishers. Printed in the Netherlands.

111

Antifungal activities of origanum oil against

Candida albicans
Vijaya Manohar,1 Cass Ingram,2 Judy Gray,2 Nadeem A. Talpur,1 Bobby
W. Echard,1 Debasis Bagchi3 and Harry G. Preuss1
1
Department of Physiology and Biophysics, Georgetown University Medical Center, Washington, DC; 2North American
Herbs and Spices, Waukeegan, IL; 3Department of Pharmacy Sciences, Creighton University School of Pharmacy and AHP,
Omaha, NE, USA

Received 27 June 2001; accepted 14 September 2001

Abstract
The antimicrobial properties of volatile aromatic oils from medicinal as well as other edible plants has been recognized since
antiquity. Origanum oil, which is used as a food flavoring agent, possesses a broad spectrum of in vitro antimicrobial activities
attributed to the high content of phenolic derivatives such as carvacrol and thymol. In the present study, antifungal properties
of origanum oil were examined both in vitro and in vivo. Using Candida albicans in broth cultures and a micro dilution method,
comparative efficacy of origanum oil, carvacrol, nystatin and amphotericin B were examined in vitro. Origanum oil at 0.25
mg/ml was found to completely inhibit the growth of C. albicans in culture. Growth inhibitions of 75% and >50% were ob-
served at 0.125 mg/ml and 0.0625 mg/ml level, respectively. In addition, both the germination and the mycelial growth of C.
albicans were found to be inhibited by origanum oil and carvacrol in a dose-dependent manner. Furthermore, the therapeutic
efficacy of origanum oil was examined in an experimental murine systemic candidiasis model. Groups of mice (n = 6) infected
with C. albicans (5 × LD50) were fed varying amounts of origanum oil in a final vol. of 0.1 ml of olive oil (vehicle). The daily
administration of 8.6 mg of origanum oil in 100 µl of olive oil/kg body weight for 30 days resulted in 80% survivability, with
no renal burden of C. albicans as opposed to the group of mice fed olive oil alone, who died within 10 days. Similar results
were obtained with carvacrol. However, mice fed origanum oil exhibited cosmetically better clinical appearance compared to
those cured with carvacrol. The results from our study encourage examination of the efficacy of origanum oil in other forms
of systemic and superficial fungal infections and exploration of its broad spectrum effect against other pathogenic manifesta-
tions including malignancy. (Mol Cell Biochem 228: 111–117, 2001

Key words: antifungal activity, in vitro, in vivo, female BALB/c mice, origanum oil, Candida albicans, carvacrol, nystatin,
amphotericin B

Introduction growth of various food spoiling organisms including the

species of Aspergillus (mycotoxinogenic filamentous fungi)
The antimicrobial properties of volatile aromatic oils from and Hansenula (industrial yeasts) [4–7]. While the oil and
edible plants have been recognized since antiquity. Origa- many of its constituents have been demonstrated to be an-
num oil, which is used as a food flavoring agent, has been tifungal in vitro against non-pathogenic yeasts [8, 9], a few
shown to possess a broad spectrum of antimicrobial activ- studies have suggested a potential therapeutic effect against
ity due to its high content of phenolic derivatives such as experimental infections in rats due to Trichophyton rubrum,
carvacrol and thymol [1–3]. Earlier studies have demon- a human dermatophytic filamentous fungus) [10]. Never-
strated the ability of origanum oil to retard and inhibit the theless, very little information is available on its compara-

Address for offprints: H.G. Preuss, Department of Physiology and Biophysics, Georgetown University Medical Center, Med-Dent Building, SE103, 3900
Reservoir Road, NW, Washington, DC 20007, USA (E-mail: preusshg@georgetown.edu)
112

tive antifungal activity on the growth and physiology of Susceptibility testing

human pathogenic yeasts or filamentous fungi either in vitro
or in vivo. Furthermore, its direct therapeutic use either in A micro-broth dilution technique was employed to determine
superficial or systemic infections due to bacteria or fungi has the susceptibility of the strains of C. albicans to oil of origa-
not been clearly established. In the present study, we exam- num and carvacrol [19, 20]. Susceptibility was expressed as
ined the antifungal properties of origanum oil and its major minimum inhibitory concentration (MIC) and minimum fun-
chemical constituent carvacrol against Candida albicans, a di- gicidal concentration (MFC). The stock solutions of origa-
morphic yeast-like fungus. C. albicans which resides as com- num oil, olive oil and carvacrol (63% v/v adjusted in olive
mensal in the mucocutaneous cavities of skin, vagina and oil) were dissolved in ethanol-Tween 80 solvent. Nystatin and
intestine of humans [11], can cause infections under altered amphotericin B were dissolved in 50% ethanol, and used as
physiological and pathological conditions such as infancy, positive controls. Solvent and media controls were also in-
pregnancy, diabetes, prolonged broad spectrum antibiotic ad- cluded for reference. The Sabouraud’s glucose (S.g) broth
ministration, steroidal chemotherapy as well as AIDS [12– containing varying amounts (logarithmic, serially and 2-fold
18]. We have demonstrated that origanum oil inhibits the diluted) of origanum oil and carvacrol and the various con-
growth of C. albicans in vitro as well as in vivo. We conclude trols were inoculated with actively dividing C. albicans cells.
that the daily oral administration of origanum oil may be highly The cultures were incubated for 24 and 48 h at 30°C on a
effective in the prevention and treatment of candidiasis. metabolic rotary shaker (200 rev/min), and the growth was
monitored both visually and colorimetrically (at 540 nM).
The minimum inhibitory concentration (MIC) was defined
as the lowest concentration required to arrest the growth of
Materials and methods the fungi at the end of 24 h of incubation. Minimum fungi-
cidal concentration (MFC) was determined by sub-culturing
Animals and treatment a 0.01 ml aliquot of the medium drawn from the culture tubes
showing no macroscopic growth at the end of 48 h of cul-
Female BALB/c mice (15–18 g) were obtained from Taconic ture on S.g. agar plates and incubated further for the appear-
Farms (Germantown, NY, USA). The animals were main- ance of yeast-like growth. The plates were scored for growth
tained in a controlled environment at 25°C with a 12 h light of the yeast colonies. The lowest concentration of the anti-
and 12 h dark cycle, and were acclimatized for 3–5 days fungal agent from which negative growth or fewer than 3
before use. The mice were housed in groups of five, fed com- colonies were recorded was considered as minimum fungi-
mercial rodent pellets and given water ad libitum through- cidal concentration (MFC).
out the experiments. The protocol for the entire investigation
was approved by the Animal Welfare Board at Georgetown
University Medical Center. Effect on yeast and mycelial forms of C. albicans

The effect of origanum oil on the formation of germ tubes

Plant oils and chemicals (filament initiation) by C. albicans was examined, by incu-
bating the blastospores with various concentrations of origa-
Origanum (P73 OreganolTM) and olive oil were provided by num oil, carvacrol and other agents in 20% egg albumin at
North American Herbs and Spices, Waukegan, IL, USA. Car- 37°C [21]. The comparative percentage inhibition of germ
vacrol, nystatin and amphotericin B were purchased from tube formation was determined by microscopic examination.
Sigma Chemical Co. (St. Louis, MO, USA). Sabouraud’s glu- The effect on the mycelial formation (filament elongation) was
cose (S.g) broth and agar media were purchased from Difco carried out by incubating the previously germinated blasto-
Laboratories (Detroit, MI, USA). All other chemicals used spores of C. albicans in the presence of varying amounts of
in this study were obtained from Sigma Chemical Co. (St antifungal agents at 37°C for an additional 24 h and 48 h. Both
Louis, MO, USA) and were of analytical grade or the high- the germination and the filament elongation were microscopi-
est commercial grade available. cally monitored. The MIC and MFC were determined as above.

Organisms Toxicity tests with mice

Standard strain of Candida albicans (ATCC No. 48274) ob- Since origanum oil is known to be pungent and corrosive, a
tained from ATCC, Fairfax, VA, USA, was grown and main- titration of the doses of origanum oil was conducted to de-
tained on S.g. agar slants. termine the tolerable dose to be given orally. Both the neat
 113

oil and the dilutions in olive oil (vehicle) were orally admin- Results
istered to the mice according to the body weight using a ball
tip gavage needle (Harvard Apparatus, South Natick, MA, Origanum oil is fungicidal to C. albicans in vitro
USA). Higher concentrations of the oil were given as single
bolus dose, while the lower concentrations were given daily In the present study, we have examined the antifungal prop-
for 7 days. The overall health of the animals such as coat erties of origanum oil and one of its major constituents, car-
color, body weight, other side effects such as scruffiness, and vacrol and thymol, both in vitro and in vivo. Using Candida
death were recorded for 14 days. At the end of the experi- albicans in broth cultures and a micro-dilution method, com-
ment, mice were sacrificed to visually examine the internal parative efficacies of origanum oil, carvacrol, nystatin and
organs for any abnormalities. amphotericin B were examined in vitro. Origanum oil at 0.25
mg/ml was found to completely inhibit the growth of C. al-
bicans in culture. Growth inhibitions of 75% and >50% were
Effect on experimental murine systemic candidiasis observed at 0.125 mg/ml and 0.0625 mg/ml level, respec-
tively (Table 1). Figure 1 provides a pictorial presentation of
A murine systemic candidiasis model developed previously the inhibitory effect. A 2-fold higher concentration of carvac-
[22], was employed to evaluate the in vivo anti fungal activi- rol was required to be fungicidal.
ties of origanum oil and carvacrol. Initially, the concentra-
tions of C. albicans needed to achieve 50% mortality of the
animals (LD50) were determined. Mice were injected i.v. with Both the germination and mycelial elongation of C.
varying doses of actively growing C. albicans in a final vol. albicans are inhibited in vitro
of 0.1 ml. Mortality of mice was monitored for 30 days. The
death of the mice due to C. albicans was confirmed by ana- Origanum oil and carvacrol both inhibit germ-tube formation
tomical evidence for organ involvement, i.e. the histological by the blastospores of C. albicans (Table 2). The production
presence of yeasts and/or pseudomycelia. Furthermore, re- of germ tubes and subsequent mycelial formation when ex-
nal burden of C. albicans was assessed by culturing an ali- posed to either serum or hen’s egg albumen is an in vitro
quot of the kidney homogenate on S.g. agar plates. The dose correlate of the in vivo tissue invasive capabilities of the
titration revealed that 2.5 × 106 C. albicans cells were needed pathogenic strains of C. albicans. Origanum oil inhibited
to kill 50% of the mice. Subsequently, batches of mice in-
jected with 5 × LD50 dose of C. albicans were used to assess
therapeutic activity. Table1. Effect of origanum oil on the growth of Candida albicans

Agent 24 h MIC 48 h MFC

(mg/ml) (mg/ml)
Treatment protocol
Origanum oil 0.125 0.25
Carvacrol 0.25 0.5
In two separate experiments, groups of mice (6 each) in- Amphotericin B 0. 0015 0.0015
fected with C. albicans (5 × LD50) were gavaged, daily with Nystatin 0. 005 0.005
origanum oil or carvacrol in 0.1 ml of olive oil for 8 days Olive oil No effect No effect
and 30 days. The amount of antifungal agents administered MIC = minimum inhibitory concentration; MFC = minimum fungicidal
was calculated based on the body weight of the mice. Con- concentration.
trol mice received either olive oil orally alone (negative
control), or olive oil orally plus amphotericin B 25 µg i.p.
(positive control). The experiments were terminated at the Table 2. Effect of origanum oil on germ tube formation and filament elon-
end of 30 days. The body weights, the disease status and gation
the overall health of the mice during the experiment were Agent Germ tube formation Filament elongation
recorded. The pathological status of the mice was deter-
mined by visual examination of the internal organs after MIC MFC MIC MFC
mg/ml mg/ml mg/ml mg/ml
their death or sacrifice at the completion of the experiment.
For histology, kidney smears were examined microscopi- Origanum oil 0. 062 0.125 0.125 0.125
cally after staining with 1% methylene blue for the presence Carvacrol 0. 125 0.125 0.125 0.25
Olive oil NE NE NE NE
of fungal elements. Renal burden of C. albicans was fur-
ther tested by culturing aliquots of kidney homogenates on NE = no effect; MIC and MFC for nystatin and amphotericin B = 2.5 and
S.g. agar plates. 5.0 µg/ml respectively.
114

Fig. 1. In vitro antifungal activity of origanum oil against Candida albicans. Exponentially growing C. albicans cells were cultured with varying concentra-
tions of antifungal agents. Aliquots from the cultures drawn at the end of 48 h of culturing were plated on to Sabouraud’s glucose agar. The plates were
further incubated for an additional 48 h for the appearance of yeast like colonies.

germ-tube production by blastospores of C. albicans in a dose- Table 3. Toxicity of origanum oil to BALB/c mice
dependent manner. The MIC required to inhibit the production
Concentration of oil* No. of mice
of germ tube formation was 0.062 mg/ml (24 h) and the fun- (mg/kg) (tested/survived)
gicidal effect (at 48 h) required double the concentration
(MFC = 0.125 mg/ml). Carvacrol exerted effects similar to Bolus dose 2600 5/0
1300 5/0
origanum oil at twice the concentrations. Origanum oil and
650 5/0
carvacrol were also found to affect filament elongation. The 325.0 5/5
mycelial forms of C. albicans were also inhibited by origa- 162.5 5/5
num oil at 0.125 mg/ml (MIC) and 0.125 mg/ml (MFC) lev-
Daily dose for 1 week 26 6/6
els, and by carvacrol at 0.125 mg/ml (MIC) and 0.25 mg/ml 52 6/6
(MFC) levels, respectively. 78 6/6
104 6/6
130 6/6
Origanum oil can protect mice from systemic candidiasis *Oil was mixed with 100 µl olive oil and gavaged.

The therapeutic efficacy of origanum oil was examined in an

experimental murine systemic candidiasis model. Toxicity Table 4. Therapeutic efficacy of origanum oil: 8-day treatment
studies to determine the least toxic dose of origanum oil that
Agent Conc. @ per No. of mice No. of mice with
can be fed safely to mice were conducted (Table 3). A single kg body wt. (mg) tested/survived renal burden#
bolus dose of 650 mg/kg body weight caused uniform mor-
tality. However, daily administration of smaller doses of ori- Origanum oil 325. 5 6/6 4/6
162. 5 6/6 6/6
ganum oil mixed with olive oil was well tolerated, showing Amphotericin B 1. 0 6/6 2/6
no apparent clinical abnormalities (Table 3). Olive oil (vehicle) 6/0* –
In the first in vivo study, the therapeutic ability of origa-
*Died within 10 days. #Renal burden+ = kidney smear+ culture+.
num oil given only for 8 days to prevent the mortality of mice
with systemic infection due to C. albicans was tested (Table
4). Groups of mice (n = 6) infected with C. albicans (5 × LD50) mg per kg body weight was found to rescue the mice from
were gavaged with varying amounts of origanum oil in a fi- mortality with 100% survival rate over the 30 days of obser-
nal vol. of 0.1 ml. An 8-day treatment with 162.5 mg and 325 vation, similar to mice that received amphotericin B (1 mg/
 115

Table 5. Therapeutic efficacy of origanum oil: 30-day treatment

Agent Conc. @ per No. of mice No. of mice with

kg body wt. (mg) tested/survived renal burden#

Origanum oil 52. 00 6/6 0/6

43. 33 6/6 0/6
31. 00 6/6 0/6
26. 00 6/6 0/6
17. 33 6/6 0/6
8. 66 6/5 0/5
Carvacrol 17. 33 6/6 0/6
8. 66 6/5 0/5
Amphotericin B 1. 0 6/6 0/6
Olive oil 6/0* 6/6

*Died within 10 days. #Renal burden+ = kidney smear+ culture+. One mouse
treated with 8.66 mg origanum oil/kg b.w. died on day 15, while one mouse
treated with 8.66 mg carvacrol/kg b.w. died on day 13. Fig. 2. In vivo effect of origanum oil in experimental murine candidiasis.
Infected mice were fed orally with varying amounts of origanum oil in 0.1
ml vol. of olive oil (vehicle). The control animals received either olive oil
kg body weight). In contrast, the control group of mice who alone or olive oil plus amphotericin B (i.p.). The body weights and the
overall health of the mice were recorded daily.
received 0.1 ml of olive oil alone died within 10 days. At the
end of 30 days, the surviving animals appeared to be clinically
healthy, without scruffiness and with regain of normal coat antimicrobial activity, including anthelmintic properties, due
luster. Internally, no abnormalities of organs were noted. Fur- to its phenolic, alcoholic and terpenoid constituents [4]. The
thermore, the kidney smears failed to demonstrate the pres- objective of the present study was to assess the antifungal
ence of either yeasts or pseudomycelia. However, the culture properties of origanum oil against C. albicans in both in vitro
of their kidney homogenates on S.g. agar revealed the pres- and in vivo. Furthermore, the comparative efficacy of origa-
ence of C. albicans. (Table 4). num oil, carvacrol, nystatin and amphotericin B was also
The results from the second experiment to determine the examined. The results clearly demonstrate that the origanum
optimal dose over a 30-day treatment course required for oil can act as a potent antifungal agent against C. albicans,
clinical cure and to completely eliminate the renal burden of and can function similar to antifungal antibiotics such as
C. albicans are shown in Table 5. A daily administration of nystatin or amphotericin B.
origanum oil as low as 8.66 mg/kg body weight (equivalent C . albicans is a harmless commensal yeast-like fungus in
of 1 µl per mouse) was found to cure 80% of the infected healthy humans, which can cause superficial as well as life-
mice. Initially all groups of infected mice, exhibited scruffi- threatening systemic infections under immune compromised
ness, loss of body weight, and lack of coat luster. In the ve- situations. C .albicans can colonize or infect virtually all body
hicle-control mice, the average body weight was found to sites because of its high adaptability to different host niches
decrease 7 days after injection from approximately 16–11 g. by the activation of appropriate sets of genes in response to
In contrast, mice receiving treatment had a gradual increase complex environmental signals [21–25]. Thus, our objec-
in body weight after injection from 15–17 to 16–19 g by the tive was to assess the possible therapeutic potential of the
end of 30 days (Fig. 2). The daily administration of equiva- commonly used origanum oil against this human dimorphic
lent amounts (8.66 and 17.33 mg/kg body weight) of carva- commensal, which can become a facultative pathogen un-
crol was found to confer similar therapeutic properties (data der altered physiological situations.
not shown). However, the overall clinical appearance of the Previous studies to assess the inhibitory effect of origanum
mice receiving origanum oil was cosmetically better than oil against twenty-five different genera of microorganisms
those receiving carvacrol, as evidenced by the improved coat including animal and plant pathogens, food poisoning and
luster and minimal scruffiness. spoilage bacteria, have demonstrated the growth inhibition
properties by origanum oil [26]. Essential oils of oregano
(Origanum vulgare) were found to inhibit the growth and
Discussion production of ochratoxin A by Aspergillus ochraceus NRRL
3174 up to 21 days in culture [27].
Aromatic herbal oils used for cooking and flavoring are in- Origanum oil has been shown to delay or inhibit the growth
creasingly claimed to have broad spectrum antimicrobial of saprophytic food spoiling fungi such as Aspergillus fla-
activities. Origanum oil has been suggested to have potent vus (mycotoxogenic) and industrial yeasts such as Hansenula
116

anoma [8]. Furthermore, origanum oil, and carvacrol have Acknowledgement

been reported to have some promising effect on rats infected
with a human dermatophytic fungus, Trichophyton rubrum This work was supported by a generous grant from North
[10]. However, the essential oils including origanum oil at American Herb and Spice located in Waukeegan, IL, USA.
100 ppm had no effect on the pseudomycelial formation by Oreganum oil was a generous gift from the North American
Candida lipolytica [10,11]. Herb and Spice, Waukeegan, IL, USA.
In this paper we have demonstrated, that origanum oil ef-
fectively inhibits the in vitro growth of C. albicans, a human
yeast-like fungus which can cause both systemic and super-
ficial infections in debilitated individuals. In addition we have References
shown that origanum oil directly inhibits germination and
filament formation (the two phases required for tissue inva- 1. Leung AY, Fostere S: Encyclopedia of Common Natural Ingredients
Used in Food, Drugs and Cosmetics, 2nd Ed. New York, John Wiley
sion) by C. albicans. Nevertheless, the novel fungicidal prop- & Sons, 1996, p 11
erty of origanum oil was of superior efficacy as compared to 2. Laredo JV, Abut M, Calvo-Torras MA: Antimicrobial activity of es-
carvacrol against the germination and mycelial elongation of sences from Labiatae. Microbios 82: 171–762, 1995
pathogenic strain of C. albicans in vitro. Furthermore, selected 3. Knobloch K, Pauli A, Iberl N, Wqeigand N, Weis HM: Antibacterial
concentrations of origanum oil demonstrated therapeutic ef- and antifungal properties of essential oil components. J Essent Oil Res
1: 119–128, 1989
ficacy comparable to the effective examined dose for am- 4. Barrata MT, Dorman HJD, Deans SG, Biondi DM, Ruberro G: Chemi-
photericin B (1.0 mg) in protecting the mice from systemic cal composition, antibacterial and antioxidative activity of laurel, sage,
candidiasis. The overall clinical appearance of the mice re- rosemary, oregano and coriander essential oils. J Essent Oil Res 10:
ceiving origanum oil was cosmetically better than those re- 618–627, 1998
ceiving carvacrol (data not shown), although this difference 5. Barrata MT, Dorman HJD, Deans SG, Biondi DM, Ruberro G: Anti-
microbial antioxidant properties of some commercial essential oils.
in appearance could possibly be attributed to the presence of Flav Frag J 13: 235–244, 1998
additional constituent phenols such as thymol, terpenes and 6. Lis-Balchin M, Deans SG: Bioactivity of selected plant essential oils
flavonoids in origanum oil [4]. It has been demonstrated that against Listeria monocytogenes. J Appl Microbiol 82: 759–762, 1997
some of these components of origanum oil have antispas- 7. Tantaoui-Elaraki A, Beraoud L: Inhibition of growth and aflatoxin
modic and antioxidant activities, in addition to their antimi- production in Aspergillus parasiticus by essential oils of selected plant
materials. J Environ Path Toxicol Oncol 13: 67–72, 1994
crobial potentials [23, 28, 29]. A comparative study of these 8. Conner DE, Beuchat, LR: Effects of essential oils from plants on growth
components may shed more light not only on the antifungal of food spoilage yeasts. J Food Sci 49: 429, 1984
potentials, but also on the pharmacological effects of each of 9. Llewellyn GC, Burkett ML, Eadie T: Potential mold growth, aflatoxin
these components. It is interesting to speculate that secondary production and antimycotic activity of selected natural spices and herbs.
infections due to C. albicans in debilitated individuals such as Assoc Anal Chem 64: 955–962, 1981
10. Adam K, Sivropoulou A, Kokkini S, Lanaras T, Arsenakis M: Antifun-
those having diabetes and HIV infection may be prophy- gal activities of Origanum vulgare subsp. Hirusutum, Mentha spicata,
lactically controlled by the daily intake of small amounts of Lavanula angustifolia, and Salvia fruticosa essential oils against hu-
oregano oil, either alone or added to a food. man pathogenic fungi. J Agricul Food Chem 46: 1739–1745, 1998
In summary, the results presented in this paper conclusively 11. Kaufman HK: Opportunistic fungal infections: Superficial and sys-
demonstrate the antifungal potential of edible origanum oil. temic candidiasis. Geriatrics 52: 50–54, 1997
12. Friedman S, Richardson SE, Jacobs SE, O’Brien K: Systemic infec-
Origanum oil is shown to be both fungistatic and fungicidal tion in extremely low birth weight infants: Short term morbidity and
to C. albicans, the human pathogenic yeast. Both the germi- neuro developmental outcome. Ped Infect Disease J 19: 499–504, 2000
nation and the pseudomycelial phase of the yeast are inhib- 13. Young GL, Jewell D: Topical treatment for vaginal candidiasis in preg-
ited in vitro. Further, the daily oral administration of as little nancy. Cochrane Datab System Rev 2: CD000225, 2000
1.0 µl of oil for 30 days completely freed 80% of the labora- 14. Rippon JW: Candidiasis and the pathogenic yeasts. In: J.W. Rippon
(ed), Med Mycol, PA, 1988, pp 532–581
tory mice from experimental systemic candidiasis, which is 15. Kennedy WA, Laurier C, Gautrin D, Ghezzo H, Pare MJL, Contandrio-
fatal if not treated. Thus, daily oral administration of origanum poulos AP: Occurrence of bad risk factors of oral candidiasis treated
oil may be highly effective in the prevention and treatment of with oral antifungals in seniors using inhaled steroids. J Clin Epidem
candidiasis. The results from our study not only encourage 53: 696–701, 2000
examination of the efficacy of origanum oil in other forms 16. Sallah S: Hepatosplenic candidiasis in patients with acute leukemia:
Increasingly encountered complication. Anticancer Res 19: 757–760,
of systemic and superficial fungal infections, but also to ex- 1999
plore its broad spectrum effect against other pathogenic 17. Jagarlamudi R, Kumar L: Systemic fungal infections in cancer patients.
manifestations including malignancies. Trop Gastroenterol 21: 3–8, 2000
 117

18. Mackenzie DWR, Cauwenberg G, Van Cutsem J, Drouhet E, Dupont B: 23. Zava DT, Delban CM, Blan M: Estrogen and bioactivity of foods, herbs
Mycoses in AIDS patients: An overview. In: H. Vanden Bossche (ed), and spices. Proc Soc Exp Biol Med 217: 369–378, 1998
Mycoses in AIDS Patients. New York, Plenum Press, 1990, p 27–33 24. Staib P, Kretschmar M, Nichterlein T, Hof H, Morschhauser J: Differ-
19. Shadomy S, Espinel-Ingroff: Susceptibility testing with antifungal ential activation of a Candida albicans virulence gene family during
Drugs. Manual Clin Microbiol 3, 1980 infection. Proc Natl Acad Sci USA 97: 6102–6107, 2000
20. Rex JH, Pfaller MA, Galgiani JN, Bartlett MS, Espinel-Ingroff A, 25. Heinig MJ, Francis J, Pappagiansis D: Mammary candidosis in lactat-
Ghannoum MA, Lancaster M, Odds FC, Rinaldi MG, Walsh TJ, Barry ing women. J Hum Lact 15: 281–288, 1999
AL: Development of interpretive breakpoints for antifungal suscepti- 26. Dorman HJ, Deans SG: Antimicrobial agents from plants: Antibacte-
bility testing: conceptual framework and analysis of in vitro–in vivo rial activity of plant volatile oils. J Appl Microbiol 88: 308–316, 2000
correlation data for flucanozole and itraconazole, and Candida infec- 27. Basilico MZ, Basilico JC: Inhibitory effects of some spice essential oils
tions. Clin Infect Dis 24: 235–247, 1997 on Aspergillus ochraceus NRRL 3174 growth and ochratoxin A pro-
21. Manohar V, Sirsi M, Ramananda Rao G: Yeasts in superficial mycoses duction. Lett Appl Microbiol 29: 238–241, 1999
I. Incidence, isolation and characterization. Indian J Med Res 63: 261– 28. Van den Broucke CO, Lemli JA: Antispasmodic activity of Origanum
265, 1975 compactum. Part 2: Antagonistic effects of thymol and carvacrol. Planta
22. Manohar V, Sirsi M, Ramananda Rao G: Yeasts in superficial mycosis Med 45: 188–190, 1982
II. Pathogenicity of Candida species to swiss albino mice. Indian J Med 29. Nakatani N: Phenolic antioxidants from herbs and spices. Biofactors
Res 64: 9–15, 1975 13: 141–146, 2000
118