Pharmaceutical Biology

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Antifungal and Antibacterial Activity of Four

Selected Mexican Medicinal Plants

Victor M. Navarro García, Gabriela Rojas, L. Gerardo Zepeda, Margarita

Aviles, Macrina Fuentes, Armando Herrera & Enrique Jiménez

To cite this article: Victor M. Navarro García, Gabriela Rojas, L. Gerardo Zepeda, Margarita
Aviles, Macrina Fuentes, Armando Herrera & Enrique Jiménez (2006) Antifungal and Antibacterial
Activity of Four Selected Mexican Medicinal Plants, Pharmaceutical Biology, 44:4, 297-300, DOI:
10.1080/13880200600715837

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Published online: 07 Oct 2008.

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Pharmaceutical Biology
2006, Vol. 44, No. 4, pp. 297–300

Antifungal and Antibacterial Activity of Four Selected Mexican

Medicinal Plants

Victor M. Navarro Garcı́a1, Gabriela Rojas1, L. Gerardo Zepeda2, Margarita Aviles3, Macrina Fuentes3,
Armando Herrera1, and Enrique Jiménez1
1
Laboratorio de Microbiologı́a, Centro de Investigaci
on Biomédica del Sur, Instituto Mexicano del Seguro Social,
Xochitepec, Morelos, México; 2Departamento de Quı́mica Organica, Escuela Nacional de Ciencias Biologicas-IPN,
México D.F., México; 3Instituto Nacional de Antropologı́a e Historia, Cuernavaca, Morelos, México

Abstract
The antifungal and antibacterial activity of 10 crude
extracts from four different species, all of them used in
Mexican folk medicine for the treatment of infectious dis-
eases, were tested in vitro for antimicrobial activity against
Staphylococcus aureus, Streptococcus faecalis, Escherichia
coli, Klebsiella pneumoniae, Salmonella typhi, Candida albi-
cans, Trichophyton mentagrophytes, and Trichophyton
rubrum. All extracts from the above plants showed some
degree of antimicrobial activity against at least two micro-
organisms tested. The strongest antibacterial activity was
found in the water extract of Hibiscus sabdariffa and the
methanol extract of Lysiloma acapulcensis, whereas the
methanol extract from Loeselia mexicana showed the best
antifungal activity against dermatophytes.
Keywords: Antibacterial activity, antifungal activity, folk
medicine, infectious diseases, medicinal plants.
Introduction
Infectious diseases represent an important problem to
health and represent one of the main causes of
morbidity and mortality worldwide (World Health
Organization, 2004). In the past years, there has been
an increasing incidence of infectious diseases due to a
growth in immunocompromised population, such as
cancer and HIV=AIDS patients. This fact coupled with
the resistance and the toxicity of several antibiotics
support the search for new drugs (Fostel & Lartey,
2000). In this sense, medicinal plants offer many possi-
bilities for discovering new molecules with antimicrobial
activity (Navarro and Delgado, 1999; Rı́os et al., 2003;
Gonzalez et al., 2004).
Mexico is a country where folk medicine still plays a
very import role in the health care system (Argueta,
1994). We are currently investigating medicinal plants
with potential biomedical uses in the treatment of infec-
tious diseases (Lozoya et al., 1992; Navarro et al., 1996;
Rojas et al., 2001). In order to extend our studies in this
field, we decided to explore the antibacterial and antifun-
gal activities of Hibiscus sabdariffa, Loeselia mexicana,
Lysiloma acapulcensis, and Miconia mexicana, which
were collected on the basis of ethnobotanical surveys
realized from the states of Morelos and Chiapas, Central
and Southern México. To our knowledge, there are no
reports describing the antimicrobial properties and
chemical constituents of these plants. Therefore, our
objective was to investigate the in vitro antimicrobial
activity of 10 extracts from the above species against
two Gram-positive bacteria, two Gram-negative bac-
teria, one yeast, and two mycelia fungi that are common
causes of several infectious diseases. The current work
will contribute to enhance our knowledge of the anti-
microbial properties of four Mexican medicinal plants,
and these results may lead to the development of new
therapeutic alternatives in phytomedicine.
Materials and Methods
Collection of plant material
Aerial parts of Loeselia mexicana and Lysiloma acapul-
censis were collected in March 2004 in their natural

Accepted: 3 March 2006

Address correspondence to: Victor M. Navarro Garcı́a, Laboratorio de Microbiologı́a, Centro de Investigaci
on Biomédica
del Sur, Instituto Mexicano del Seguro Social, Argentina 1, 62790 Xochitepec, Morelos, México. Tel.=Fax: þ777 361 21 55;
E-mail: vmnavg@yahoo.com.mx

DOI: 10.1080/13880200600715837 # 2006 Taylor & Francis Group, LLC

298 V.M. Navarro Garcı́a et al.
Table 1. List of medical plant used to evaluate their antimicrobial properties.
Species (family)
(voucher specimen number) Local name
Lysiloma acapulcensis Tepejuaje
(Kunth) Benth.
(Fabaceae)
(INHAM 2002)
Loeselia mexicana (Lam) Espinosilla
Brand
(Palemoniaceae)
(INAHAM 2017)
Miconia mexicana Itswa
(Bonpl.) Naudin
(Melastomataceae)
(INHAM 2018)
Hibiscus sabdariffa L. Flor de Jamaica
(Malvaceae)
(IMSSM-14290)
habitat from different regions in Morelos State, México.
Miconia mexicana was collected in January 2001 in
Oxchiuc, Chiapas State, México. Voucher specimens
were prepared and authenticated by M. en C. Margarita
Aviles and M. en C. Macrina Fuentes and were deposited
at the herbarium of Instituto Nacional de Antropologı́a e
Historia de Morelos (INAHM) in the city of Cuerna-
vaca, Morelos State. Hibiscus sabdariffa was collected
in February 2004 from cultivate grown controlled at
Centro de Investigaci on Biomédica del Sur, Xochitepec,
in Morelos. This plant was botanically authenticated
by M. en C. Abigail Aguilar at the herbarium of the
Instituto Mexicano del Seguro Social (IMSSM). The
voucher specimen numbers are given in Table 1. Tra-
ditional practices by herbalists in Morelos State provided
the basis for selecting the part of the plant to be tested.
Preparation of extracts
Loeselia mexicana, Lysiloma acapulcensis, and Miconia
mexicana were dried at room temperature for a period
of 2 weeks and then milled into fine powder, which
was successively extracted with n-hexane, dichloro-
methane, and methanol (100 g=1500 ml) at room tem-
perature. Each solvent was replaced three-times with
fresh solvent, remaining in contact with the plant
material 24 h each time. After filtration, the extracts were
concentrated to residue by removing the solvents in a
rotavapor at 40
C. Roughly powdered calyces (100 g)
from Hibiscus sabdariffa were macerated in 1000 ml of
distilled water by shaking at room temperatures for
24 h twice. The extract was filtered and freeze-dried.
The yield of the extracts was quantified, and the obtained
material was protected from direct light and stored at less
than 4
C until its use.
Popular uses (route of administration) (references)
For wounds, gastrointestinal disorders, bladder infections
(oral and topics) (Argueta et al., 1994; Avilés and Su
arez,
1994; Monroy and Castillo, 2000, Avilés 2001)
For skin diseases, diarrhea, respiratory disease, fever,
dandruff, hair care, avoid loss of hair (oral and topic)
(Argueta et al., 1994; Aguilar et al., 1994)
Anti-inflammatory, anthelmintic, cough treatment (oral
and topic) (Berlin et al., 1990)
Intestinal pain, diuretic, fever, and reduce blood pressure
(oral) (Argueta et al., 1994; Monroy and Castillo, 2000)
Antibacterial activity
The bacteria used for the antibacterial assays were pur-
chased from American Type Culture Collection (ATCC,
Rockville, MD, USA). The following strains were used:
Staphylococcus aureus ATCC 6358; Streptococcus faeca-
lis ATCC 10231; Escherichia coli ATCC 8937; Klebsiella
pneumoniae ATCC 13883, and Salmonela typhi ATCC
06539.
The antibacterial activity was measured by determin-
ing the minimal inhibitory concentration (MIC) and
was carried out by employing the agar dilution method
(Rı́os et al., 1988). The crude extracts (n-hexane, dichlor-
omethane, and methanol) were dissolved in dimethyl
sulfoxide (DMSO; 2% v=v); to obtain a concentration
of 40 mg=ml. Dilutions were prepared from these solu-
tions to obtain final concentrations of 8.0 to 0.5 mg=ml.
The inoculum for each organism was prepared from
cultures containing 108 colony-forming units (CFU)=ml.
The diluted (1:20) inoculum was applied as a spot by
means of a calibrated loop to deliver 0.002 ml, resulting
in a spot inoculum covering a circle of 5–8 mm diameter
and containing 104 CFU. The plates were incubated for
24 h at 37
C. Gentamicin (2.5–120 mg=ml) (Sigma) was
used as reference standard. Observations were performed
by duplicate, and results are expressed as the lowest con-
centration of plant extract able to produce a complete
suppression of colony growth on agar (minimum inhibi-
tory concentration; MIC).
Antifungal activity
The following three strains were used as test fungi: Try-
chophyton mentagrophytes ATCC 28185, Trychophyton
rubrum ATCC 28188, and Candida albicans ATCC
 Antimicrobial activity of Mexican plants 299

10231. The filamentous fungi were maintained on potato
dextrose agar (PDA; Merck, Germany) at 27
C. Sabour-
aud glucose agar (SGA; Merck, Germany) was used to
keep up the yeast and as assay medium.
The antifungal assay was performed by the agar dilution
method using Petri dishes (Falcon, USA) (Rahalison
et al., 1994; Gadhi et al., 2001). The stock solution of
extracts and reference compounds, nystatin (Merck,
Germany) and miconazole (Sigma), were two-fold serial
diluted yielding concentrations in the range from 8.0 to
0.125 mg=ml and from 128 to 1 mg=ml, respectively. Final
concentrations of DMSO in the test were less than 2%
(v=v). Final inoculum of 105 cell=ml for Candida albicans
and 106 spore=ml for filamentous fungi was placed on
top of the solidified agar with a loop calibrated to deliver
0.005 ml. Experiments were carried out in duplicate
and incubation at 29
C. The fungal growth was checked,
first in control plates prepared without any test sample,
after 24, 48, and 72 h, depending on the incubation
period required for a visible growth: 24 h for
Candida albicans, 24 h for Aspergillus niger, and 72 h for
dermatophytes.
Results and Discussion
Table 1 shows the botanical name, voucher specimen
numbers, local name, and popular uses of the studied
plants. In Table 2, the plant part used, the percentage
yield, and the obtained MIC values of the corresponding
extracts are summarized. Considering that in this screen-
ing only crude extracts were tested, extracts with MIC
values of 8.0 mg=ml or below were considered active.
The obtained results show that all the tested plants
were active against at least two of the assayed fungi.
As antifungal, the most interesting were the methanol
extracts of Lysiloma acapulcensis (C. albicans MIC ¼ 2.0,
T. mentagrophytes MIC ¼ 1.0, and T. rubrum
MIC ¼ 1.0 mg=ml) and Loeselia mexicana (C. albicans
MIC ¼ 4.0, T. mentagrophytes MIC ¼ 0.250, and
T. rubrum MIC ¼ 0.250 mg=ml). Also, the dichloro-
methanol and hexane extracts of L. mexicana showed
an important activity against dermatophytes (Table 2).
Concerning the antibacterial properties, the most
interesting results were obtained from methanol extracts
of Lysiloma acapulcensis, Miconia mexicana, and
Hibiscus sabdariffa (water extract), with the last extract
being the most active against the Gram-positive and
Gram-negative bacteria. Thus, MIC of 0.5 mg=ml was
found against S. aureus and S. faecalis, while a MIC of
1.0 mg=ml was found against E. coli, K. pneumoniae,
and S. typhi.
These screenings showed that all tested plant extracts
possess interesting antimicrobial properties. This
explains the use of these plants in folk medicine for the
treatment of various illnesses whose symptoms might
involve infectious diseases. Consequently, in order to
elucidate the responsible active compound, we have
currently started a bioguided fractionation of the above
extracts.

Table 2. Minimal inhibitory concentration (MIC, mg=ml) of the organic crude extract.

Extract MIC
Used plant yield
Species parta Extractb (%)c Sad Sf Ec Kn St Ca Tm Tr

H 2.6 4.0 4.0 > 8.0 > 8.0 > 8.0 > 8.0 > 8.0 > 8.0
Lysiloma acapulcensis B D 3.5 1.0 2.0 4.0 4.0 2.0 4.0 2.0 2.0
M 4.9 1.0 2.0 4.0 2.0 2.0 2.0 1.0 1.0
H 3.1 0.5 2.0 4.0 2.0 4.0 8.0 4.0 4.0
Miconia mexicana L D 5.2 > 8.0 > 8.0 > 8.0 > 8.0 > 8.0 > 8.0 2.0 4.0
M 9.1 0.5 2.0 4.0 2.0 4.0 4.0 2.0 2.0
H 2.9 2.0 4.0 > 8.0 > 8.0 > 8.0 > 8.0 1.0 1.0
Loeselia mexicana S D 3.7 2.0 2.0 > 8.0 > 8.0 > 8.0 > 8.0 0.5 0.5
M 5.1 > 8.0 > 8.0 > 8.0 > 8.0 > 8.0 4.0 0.25 0.25
Hibiscus sabdariffa C W 38.0 0.5 0.5 1.0 1.0 1.0 8.0 4.0 4.0
Gentamycin 0.002 0.002 0.008 0.008 0.008 0.008 NT NT
Myconazol NTe NT NT NT NT NT 0.008 0.008
Nystatin NT NT NT NT NT 0.008 NT NT
a
Plant part tested: B, bark; C, calyces; L, leaf; S, stem.
b
Extract: D, dichloromethane; H, hexane; W, water extract; M, methanol.
c
Based on dry starting material.
d
Microbial species: Sa, Staphylococcus aureus; Sf, Streptococcus faecalis; Ec, Escherichia coli; Kn, Klebsiella pneumoniae; St,
Salmonella typhi; Ca, Candida albicans; Tm, Trichophyton mentagrophytes; Tr, Trichophyton rubrum.
e
Not tested.
300 V.M. Navarro Garcı́a et al.
Acknowledgments
We want to thank Myrna and Francois Mages for tech-
nical assistance. This investigation was partially sup-
ported by project IMSS-FOFOI: 2005=1=I=066.
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