Journal of Ethnopharmacology 109 (2007) 435–441

Evaluation of the flora of Northern Mexico for in vitro antimicrobial and

antituberculosis activity夽
G.M. Molina-Salinas a,b , A. Pérez-López a , P. Becerril-Montes b , R. Salazar-Aranda a ,
S. Said-Fernández b , N. Waksman de Torres a,∗
a Departmento de Quı́mica Analı́tica, Facultad de Medicina, U.A.N.L. P.O. Box 2316, Sucursal Tecnológico, 64841 Monterrey, N.L., México
b División de Biologı́a Celular y Molecular, Centro de Investigación Biomédica del Noreste, Instituto Mexicano del Seguro Social,

Monterrey, N.L., CP 64720, México

Received 1 February 2006; received in revised form 8 August 2006; accepted 16 August 2006
Available online 23 August 2006

Abstract
The aim of the present study was to evaluate the potential antimicrobial activity of 14 plants used in northeast México for the treatment
of respiratory diseases, against drug-sensitive and drug-resistant strains of Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus
influenzae type b and Mycobacterium tuberculosis. Forty-eight organic and aqueous extracts were tested against these bacterial strains using
a broth microdilution test. No aqueous extracts showed antimicrobial activity, whereas most of the organic extracts presented antimicrobial
activity against at least one of the drug-resistant microorganisms tested. Methanol-based extracts from the roots and leaves of Leucophyllum
frutescens and ethyl ether extract from the roots of Chrysanctinia mexicana showed the greatest antimicrobial activity against the drug-resistant
strain of Mycobacterium tuberculosis; the minimal inhibitory concentration (MIC) were 62.5, 125 and 62.5 ␮g/mL, respectively; methanol-based
extract from the leaves of Cordia boissieri showed the best antimicrobial activity against the drug-resistant strain of Staphylococcus aureus (MIC
250 ␮g/mL); the hexane-based extract from the fruits of Schinus molle showed considerable antimicrobial activity against the drug-resistant strain
of Streptococcus pneumoniae (MIC 62.5 ␮g/mL). This study supports that selecting plants by ethnobotanical criteria enhances the possibility of
finding species with activity against resistant microorganisms.
© 2006 Elsevier Ireland Ltd. All rights reserved.

Keywords: Mycobacterium tuberculosis; Antimicrobial activity; Mexican plants; Alamar Blue; Plant extracts

1. Introduction The emergence and spread of drug-resistant Mycobacterium

Tuberculosis and pneumonia are two of the most impor-
tant diseases worldwide. Current epidemiological evidence sug-
gests that one third of the world’s population is infected with
Mycobacterium tuberculosis, eight million cases emerge annu-
ally and three million deaths per year are directly attributable to
infection with this bacillus (WHO, 2002). Pneumonia is also a
common infectious disease, having a high annual mortality rate
worldwide (Thomson and Miller, 2003).
The worldwide increase in incidence and mortality from
tuberculosis prompted the WHO to declare this disease a global
emergency in the early 1990s (WHO, 1994).
夽 The contribution of both groups to the present study was equally important.
∗ Corresponding author. Tel.: +52 8183294185; fax: +52 8186758546.
E-mail address: nwaksman@fm.uanl.mx (N.W.d. Torres).
tuberculosis strains, especially multidrug-resistant (MDR)
strains resistant to isoniazid and rifampin, decrease treatment
efficacy and threaten advances in the control of tuberculosis.
The emergence of drug-resistant strains of Mycobacterium
tuberculosis has led to increased pressure on current chemother-
apy regimes. In México, 13,526 new cases of pulmonary tuber-
culosis were reported in 2004 (Secretarı́a de Salud de México,
2005). In Monterrey, Nuevo León, 186 strains were isolated from
patients of the Hospital Universitario José Eleuterio González
between 1996 and 1998. Of these strains 32% were resistant to
at least one antituberculosis drug and 18% were MDR (Yang et
al., 2001). The isolation of 65 strains from patients in another
hospital in Monterrey revealed that 47.7% of strains were resis-
tant to the first line of antituberculosis drugs and 52.3% were
resistant to at least one medication (Said-Fernández et al., 2001).
The emergence of methicillin-resistant Staphylococcus
aureus (MRSA) strains in the 1960s, and more recently the

0378-8741/$ – see front matter © 2006 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.jep.2006.08.014
436 G.M. Molina-Salinas et al. / Journal of Ethnopharmacology 109 (2007) 435–441

Table 1
Plants species from northeast México screened for antimicrobial activity
Plant (family) Local name Popular use Collection site Voucher specimen References
UNL

Ceanothus coeruleus Lag. Conchinchilla Fever, angina C 024099 Dı́az (1976)

(Rhamanceae)
Chrysactinia mexicana Gray
(Compositae)
Clematis drummondii T and
G. (Ranunculaceae)
Colubrina greggii Wats.
(Rhamanceae)
Cordia boissieri A. DC.
(Borraginaceae)
Eupatorium odoratum L.
(Compositae)
Heliotropium angiospermum
Murr (Borraginaceae)
Leucophyllum frutescens
(Berl.) I.M. Johnst.
(Escrophulariaceae)
Porlieria angustifolia
(Zigophylaceae)
Phyla nodiflora (L.) Greene.
(Verbenaceae)
Sambucus mexicana Presl.
(Capripholiaceae)
Schinus molle L.
(Anacardiaceae)
Scutellaria elliptica Muhl
(Labiatae)
Smilax bona nox L.
(Smilacaceae)
Hierba de San Nicolás Respiratory affections
Barbas de Chivo Disinfectant, antibiotic
Guajolote Fever, antibiotic
Anacahuita Tuberculosis, cough
Crucita Chest complaints, pulmonary
affections
Cola de escorpión Pulmonary affections,
pharyngitis
Cenizo Tuberculosis, bronchitis
Guayacán Tuberculosis, cough, cold
Cola de Caballo Fever, boil
Sauco Chest complaints, cough,
cold, fever, throat pain
Pirul Tuberculosis, bronchitis,
cough, cold, fever,
expectorant, asthma
Hairy scullkap Cough, expectorant, fever,
cold
Zarzaparrilla Cough, fever, leprae, cold,
expectorant,
C 024102 Adame and Adame
(2000), Cantrell et al.
(1998)
G 024164 Adame and Adame
(2000), Cantrell et al.
(1998)
S 024169 Dı́az (1976)
M 024167 Adame and Adame
(2000)
S 024100 Cantrell et al. (1998)
G 024172
M 024165 Adame and Adame
(2000)
A 024173 Adame and Adame
(2000)
S 024168
S 024171 Adame and Adame
(2000)
C 024166 Adame and Adame
(2000)
C 024104
S 024098

C: Arteaga, Coahuila; M: Monterrey, Nuevo León; S: Santiago, Nuevo León; G: Garcı́a, Nuevo León; A: Agualeguas, Nuevo León.

emergence of MRSA strains resistant to vancomycin, has
become a cause for public health concern because vancomycin
is one of the few effective antibiotics available to treat infections
produced by MRSA (Robinson and Enright, 2003). The number
of strains of Streptococcus pneumoniae resistant to penicillin,
other ␤-lactams and vancomycin is increasing (Novak et al.,
1999). Even though the phenotypic resistance in Haemophilus
influenzae has not changed dramatically, some strains of this
species that produce ␤-lactamase have already been described
(Thornsberry et al., 1999).
The increase in incidence of new and reemerging infectious
diseases, has led to the need for new antimicrobial compounds
that have diverse chemical structures and novel mechanisms
of action. Natural products or their semisynthetic derivatives,
provide novel examples of such anti-infective drugs (Copp,
2003). In México, like other developing countries, traditional
medicine is an important source of products for treating com-
mon infections. About 25% of the Mexican population depends
exclusively on the use of medicinal plants.
Some plants that grow in south, central and northwest
México, and are used in the treatment of respiratory diseases,
reportedly have antibacterial activity (Encarnación et al., 1998;
Rojas et al., 2001; Jimenez-Arellanes et al., 2003). However,
there have been no systematic studies conducted of plants from
northeast México that are effective in treating respiratory dis-
eases. The aim of the present study was to evaluate 14 plants
that grow in northeast México to identify those that could serve
as good candidates for the development of new medication or
phytomedicines. The plants were mainly selected on base of
their ethnomedicinal use against respiratory infections or con-
ditions related to them. Biological assays were then performed
with drug-sensitive and drug-resistant bacterial strains.
2. Materials and methods
2.1. Plant material
All plants were collected from the field and selected because
of their traditional use as a treatment for tuberculosis, fever,
cough, blood in the sputum or other pulmonary diseases. Col-
lections were carried out between October 2000 and July 2002
from different regions in the states of Nuevo León and Coahuila
(Table 1). Different parts of the plants were collected and treated
independently. Voucher specimens for each plant were deposited
in the herbarium of the Facultad de Ciencias Biológicas, Uni-
versidad Autónoma de Nuevo León.
 G.M. Molina-Salinas et al. / Journal of Ethnopharmacology 109 (2007) 435–441
2.2. Reagents and antibiotics
The antibiotics rifampin, oxacillin, vancomycin, ofloxacin
(Sigma–Aldrich Chemical Co., St. Louis, MO, USA), and
cephalothin (Eli Lilly, México D.F., México) were used as con-
trols. Rifampin and ofloxacin were dissolved in 100% DMSO
(Sigma–Aldrich) and 0.1N NaOH, respectively, and diluted
with water to a final concentration of 1 mg/mL. Vancomycin
and cephalothin were dissolved in water at a concentration of
2.56 mg/mL. Standards were dispensed into 0.5 mL aliquots and
stored at −70 ◦ C until use.
2.3. Extraction procedure
Air-dried, finely powdered plant material (100 g) was
extracted by direct maceration with methanol (3 × 600 mL) for
2 h at room temperature. Hexane was used instead of methanol
for extraction of Schinus molle and Chrysanctinia mexicana was
extracted with diethyl ether (3 × 600 mL, room temperature)
before extraction with methanol.
Aqueous extracts were prepared by maceration of residue
from the methanol extractions using distilled water. Organic
extracts were filtered and evaporated to dryness under low pres-
sure at 38 ◦ C. The aqueous extracts were lyophilized. The dif-
ferent extracts were refrigerated at 4 ◦ C until tested.
2.4. Microorganisms
The following strains were used in the present study:
Mycobacterium tuberculosis H37 Rv ATTC 27294 (SMtb) and
CIBIN/UMF15:99 (RMtb). The SMtb strain is sensitive to all
five first-line antituberculosis drugs (streptomycin, isoniazid,
rifampin, ethambutol and pyrazinamide) and the RMtb strain,
which was isolated, identified and characterized in the mycobac-
teriology laboratory of the Centro de Investigación Biomédica
del Noreste, Instituto Mexicano del Seguro Social (Monter-
rey, Nuevo León, México), was obtained from a patient with
advanced pulmonary tuberculosis. It is also resistant to all the
aforementioned antituberculosis drugs. Staphylococcus aureus
ATTC 25923 (SSa) is sensitive to oxacillin; Staphylococcus
aureus IMSS-NL/HE25:01, which is resistant to oxacillin (RSa),
was isolated from a patient with systemic infection and charac-
terized in the clinical laboratory of the Unidad Médica de Alta
Especialidad no. 25, Instituto Mexicano del Seguro Social (Mon-
terrey, Nuevo León, México). Haemophilus influenzae type b
InDRE-90-CCH-02 (SHi) and Haemophilus influenzae type b
InDRE 49247 (RHi) are sensitive and resistant, respectively,
to ampicillin, which were obtained from the Instituto Nacional
de Diagnóstico y Referencia Epidemiológicos (InDRE, México
D.F., México). Streptococcus pneumoniae InDRE 24-CCpn-02
(R1Sp) and Streptococcus pneumoniae InDRE 49619 (R2Sp),
are resistant to oxacillin and sensitive to vancomycin.
2.5. Determination of antimicrobial activity
2.5.1. Preparation of inocula for biological assays
Mycobacterium tuberculosis strains were cultured at 37 ◦ C
in Middlebrook 7H9 broth supplemented with 0.2% glyc-
437
erol (Sigma–Aldrich) and 10% OADC enrichment (Oleic
acid–Albumin–Dextrose–Catalase; Becton Dickinson) until the
logarithmic phase of growth was reached. The inoculum for the
assay was prepared by diluting a logarithmically growing culture
to match the McFarland 1 turbidity standard and then further
diluting this to 1:50 with Middlebrook 7H9 broth to obtain a
concentration of 6 × 106 colony forming units/mL (Franzblau
et al., 1998). The working suspension was prepared just before
inoculation of the microplate.
Staphylococcus aureus strains were inoculated on Mueller–
Hinton Agar (MHA) plates (Becton Dickinson) and incubated
overnight at 37 ◦ C. Four to five colonies were transferred to
3 mL fresh MHB and maintained at 37 ◦ C for 3–4 h until growth
matched the McFarland 0.5 turbidity standard. The working
suspension was a 1:50 dilution of this culture in fresh broth
(NCCLS, 2002). Streptococcus pneumoniae and Haemophilus
influenzae strains were cultured in Petri dishes containing blood
agar and chocolate agar (Becton Dickinson), respectively. Plates
were incubated overnight at 37 ◦ C and suspensions were pre-
pared by transferring colonies to HTM until the turbidity of the
0.5 McFarland standard was reached. The Streptococcus pneu-
moniae working suspension was further diluted to 1:50 with
CAMHB-LHB and Haemophilus influenzae was diluted to 1:50
with HTM.
2.5.2. Preparation of extracts for biological assays
The organic extracts for Mycobacterium tuberculosis bioas-
says were prepared at a concentration of 2 mg/mL in 20% DMSO
in Middlebrook 7H9 (Becton Dickinson and Co., Sparks MD,
USA) broth. For bioassays of the other microorganisms, organic
extracts were prepared at a concentration of 2 mg/mL in 20%
DMSO in culture medium: Mueller–Hinton Broth (MHB; Bec-
ton Dickinson) for Staphylococcus aureus; Haemophilus test
medium (HTM, Becton Dickinson) for Haemophilus influenzae,
prepared as reported previously (NCCLS, 2002) and Cation-
Adjusted Mueller–Hinton Broth supplemented with 5% lysed
horse blood (CAMHB-LHB; Becton Dickinson) for Strepto-
coccus pneumoniae. These solutions were sterilized by filtra-
tion using 13 mm diameter PTFE acrodiscs (0.22 ␮m pore size;
Millipore Co., Bedford, MA, USA). The aqueous extracts for
testing against Mycobacterium tuberculosis were dissolved in
Middlebrook 7H9 broth at a concentration of 2 mg/mL. For the
other bacteria, aqueous extracts were prepared in the appropriate
medium (MHB, HTM or CAMHB-LHB) at a concentration of
2 mg/mL. All solutions were sterilized by filtration using 13 mm
diameter nylon acrodiscs (0.22 ␮m pore size, Pall-Gelman Lab,
Ann Arbor, MI, USA).
2.5.3. Susceptibility screening
The antibacterial activity of plant extracts against Mycobac-
terium tuberculosis strains was tested using the microplate
Alamar Blue assay (MABA; Franzblau et al., 1998) modified
by Molina-Salinas et al. (2006). The concentrations for plant
extracts ranged from 500.000 to 15.625 ␮g/mL. Final antibi-
otic concentrations used were 2.000–0.062 ␮g/mL rifampin and
16.000–0.500 ␮g/mL ofloxacin.
438 G.M. Molina-Salinas et al. / Journal of Ethnopharmacology 109 (2007) 435–441

Staphylococcus aureus and Haemophilus influenzae strains Table 3

were tested with a microdilution assay according to the National
Committee for Clinical Laboratory Standards (NCCLS, 2002;
Salvat et al., 2004) with minor modifications. The culture
medium varied according to the microorganism being tested.
MHB was used for sensitivity testing of Staphylococcus aureus
and HTM medium was used for Haemophilus influenzae.
Microdilution tests were performed in sterile flat-bottom 96-
well polystyrene microplates covered with a low evaporation lid.
Outer-perimeter wells were filled with 200 ␮L sterile water and
100 ␮L of the corresponding medium was added to each well.
A 100 ␮L volume of the extract sample was transferred to the
first well of each row, a serial two-fold dilution was performed
and the remaining 100 ␮L was discarded. Seven samples and
two antimicrobial drug controls, oxacillin or vancomycin and
cephalothin (range 64–0.4 ␮g/mL), were included in each plate.
Five additional wells were used as growth controls, where no
drugs were added, and culture medium was added to only one
well. Plates were placed into plastic bags, incubated at 37 ◦ C
for 24 h and bacterial growth then visually examined and 32 ␮L
of a 1.6:1 Alamar Blue solution in 0.85% NaCl was added to
each well. Plates were let stand at 37 ◦ C for 1 h. Color change of
the Alamar Blue solution from blue to pink confirmed bacterial
growth. The MIC was defined as the lowest extract concentration
that prevented bacterial growth.
The effect of plants extracts against Streptococcus pneumo-
niae strains were carried out using the microdilution method
(NCCLS, 2002). The MIC was defined by turbidity and a yel-
low color in wells after incubation indicated bacterial growth.
All biological assays were developed at least by duplicate.
3. Results and discussion
Most of the plants used in this study were selected based on
interviews with healers and individuals who had experience in
traditional medicine.
Table 1 shows the botanical name, local name, popular use
and voucher specimen for each of the selected plant species.
Tables 2–5 summarize the MIC values obtained with extracts
from these plants against Mycobacterium tuberculosis, Staphy-
lococcus aureus, Haemophilus influenzae and Streptococcus
Methanolic extracts active against Staphylococcus aureus
Plant Part MIC (␮g/mL)
SSa RSa
Ceanothus coeruleus R 500 500
Cordia boissieri L 250 250
Cephalothin 6.4 >64
Oxacillin 3.9 >64
L: leaves; R: roots. SSa: Staphylococcus aureus ATCC 25923 sensitive strain;
RSa: Staphylococcus aureus IMSS/HE25:01 resistant strain.
pneumoniae, respectively. In total, 48 extracts were tested. No
aqueous extracts showed antimicrobial activity and so are not
reported. Chrysanctinia mexicana was extracted with ether pre-
viously to the extraction with methanol, on account of its large
content of volatile components, especially essential oils; Schinus
molle fruits were extremely oily, therefore a hexanic extract was
also obtained. Extracts were considered active if they gave a MIC
≤125 ␮g/mL against some strains of Mycobacterium tubercu-
losis and a MIC ≤500 ␮g/mL against any of the other strains
tested.
All bacteria tested were inhibited by at least one extract.
The microorganism most sensitive was Streptococcus pneumo-
niae. DMSO showed some inhibitory effect at a concentration
of 6.25%, but the highest concentration of DMSO used was 5%.
Plants that showed significant antituberculosis activity were
Leucophyllum frutescens and Chrysanctinia mexicana (Table 2).
As shown in Table 3, two extracts were active against Staphy-
lococcus aureus, the most potent being the leaf extract from
Cordia boissieri. Eight extracts belonging to five plant species
were active against Haemophilus influenzae SHi or RHi strains
(Table 4), but no extracts showed a MIC lower than 500 ␮g/mL.
Both resistant Streptococcus pneumoniae strains were sensitive
to the greatest number of extracts (Table 5). The most active
extracts were the leaf extract of Cordia boissieri and the fruit
extract of Schinus molle.
Table 4
Organic extracts active against Haemophilus influenzae
Plant Part MIC (␮g/mL)

SHi RHi
Table 2
Organic extracts active against Mycobacterium tuberculosis Ceanothus coereleus R 500 500
Chrysactinia mexicana Fla NA 500
Plant Part MIC (␮g/mL) Ra 500 500
SMtb RMtb Cordia boissieri L NA 500
Phyla nodiflora L NA 500
Chrysactinia mexicana Ra 62.5 62.5
Schinus molle B NA 500
Leucophyllum frutescens L NA 125
Frb NA 500
R 62.5 62.5
R 500 500
Schinus molle Frb 125 NA
Isoniazid 0.062 3.125 Cephallotin 4 4
Rifampin 0.062 100 Oxacillin 4 4

L: leaves, Fr: fruits, and R: roots. SMtb: Mycobacterium tuberculosis ATCC
27294 sensitive strain; RMtb: Mycobacterium tuberculosis CIBIN/UMF15:99
MDR strain. NA: non active, criterion in text.
a All are methanolic extracts except: ethyl ether extract.
b All are methanolic extracts except: hexanic extract.
L: leaves, Fl: flowers, Fr: fruits, R: roots, and B: bark. SHi: Haemophilus influen-
zae 90-CCH-02 strain; RHi: Haemophilus influenzae ATCC 49247 resistant
strain to penicillin, methicillin. NA: non active, criterion in text.
a All are methanolic extracts except: ethyl ether extract.
b All are methanolic extracts except: hexanic extract.
 G.M. Molina-Salinas et al. / Journal of Ethnopharmacology 109 (2007) 435–441
Table 5
Organic extracts active against Streptococcus pneumoniae
Plant Part MIC (␮g/mL)
R1Sp
Ceanothus coereleus R 500
Chrysactinia mexicana L NA
Fla NA
Fl NA
Ra 500
R NA
Colubrina greggi R 500
Cordia boissieri L 125
Fl 500
Fr 500
R NA
Eupatorium odoratum L 250
Fl 500
R NA
Leucophyllum frutescens L 500
Fl NA
R 500
Phyla nodiflora L NA
Sambucus mexicana Fl/Fr 500
Schinus molle L 500
Fl 250
B NA
Frb 62.5
R 500
Scutellaria elliptica Fl/Fr 250
R 500
Cephalotin 1 4
Oxacillin 1 8
L: leaves, Fl: flowers, Fr: fruits, R: roots, and B: Barks. R1Sp: Streptococcus
pneumoniae 90-49619 resistant strain; R2Sp: Streptococcus pneumoniae 90-
CCpn-02 resistant strain. NA: non active, criterion in text.
a All are methanolic extracts except: ethyl ether extract.
b All are methanolic extracts except: hexanic extract.
We found that the microcolorimetric assay based on the use
of Alamar Blue is an excellent method for confirming results
obtained from screening crude plant extracts, not only when
testing them against Mycobacterium tuberculosis, as already
reported, but also against other microorganisms. However, it
is necessary that the inocula and incubation times be care-
fully standardized. This was done for testing the activity of
plant extracts against Staphylococcus aureus and Haemophilus
influenzae. However, Alamar Blue could not be used for Strep-
tococcus pneumoniae testing due to the color of the medium,
which interferes with visual observation of the change in color
of the Alamar Blue. In this case, a change in the color of the
medium was observed when the bacteria grew (NCCLS, 2002).
MIC values of up to 125 ␮g/mL for Mycobacterium tubercu-
losis and 500 ␮g/mL for Staphylococcus aureus, Streptococcus
pneumoniae and Haemophilus influenzae were used as criteria
for determining antibacterial activity. Although some consen-
sus on these values can be found in the literature concerning
Mycobacterium tuberculosis, in the case of other microorgan-
isms tested, there does not appear to be a clear criterion for
439
determining the lower concentrations of plant extracts that can
be considered as having an adequate antibacterial activity. Some
extracts having an MIC as high as 5 mg/mL (Rojas et al., 2001),
R2Sp
1 mg/mL (Salvat et al., 2004) and 0.5 mg/mL (Salvat et al., 2001)
have been considered active. In a recent review Rios suggested
500 to avoid experiments with quantities higher than 1 mg/mL (Rios
500
500
and Recio, 2005).
500 Both Mycobacterium tuberculosis strain H37 Rv and a clin-
250 ical isolate of Mycobacterium tuberculosis were used in this
500 study. Our rationale for this was that the first strain is used
500 around the world as a standard and the second strain is resistant
125 to all five first-line antituberculosis drugs and could provide an
250 opportunity to identify new compounds effective against MDR
250
strains. The emergence and spread of MDR strains around the
250
world is making the control of tuberculosis difficult (Rattan et
250 al., 1998). Our results support the potential for identifying new
500
500
compounds effective against MDR strains because we identi-
fied three plant species (Chrysanctinia mexicana, Leucophyllum
500
frutescens and Schinus molle) that have acceptable antitubercu-
500
250 losis activity against both the sensitive and the resistant strains
(62.5–125 ␮g/mL).
500
500
Streptococcus pneumoniae and Haemophilus influenzae were
250 included in the study because these species are some of the most
250 common bacterial agents of pneumonia. Additionally, we chose
250 Staphylococcus aureus as it is a frequent nosocomial pathogen
62.5 causing a wide variety of diseases, including pneumonia. We
250
included sensitive strains as well as one strain of Staphylococcus
500 aureus that is resistant to oxacillin, one strain of Haemophilus
500
influenzae resistant to ampicillin and two strains of Strepto-
coccus pneumoniae resistant to oxacillin. Treatment of patients
infected with these strains is difficult because bacteria are resis-
tant to a variety of penicillins (Jacobs et al., 1999; Strahilevitz
and Rubinstein, 2002). The plants showing the best activity
against these microorganisms were Cordia boissieri, Eupato-
rium odoratum and Schinus molle.
It is noteworthy that, except in the case of Haemophilus
influenzae, some extracts were more active against resistant
strains than sensitive strains. This was the case for Leucophyl-
lum frutescens against Mycobacterium tuberculosis and Cordia
boissieri, Chrysanctinia mexicana, Phyla nodiflora, and Schinus
molle against Streptococcus pneumoniae. This could mean that
these plants produce molecules directed against metabolic tar-
gets that are more susceptible in resistant strains than in sensitive
strains, but this possibility needs to be further investigated.
Leucophyllum frutescens (cenizo de Monterrey) is a perennial
medicinal herb, which grows along the Sierra Madre Oriental
in northeast México, from Nuevo León to Hidalgo. There are
reports of its use in traditional medicine to treat lung complaints
including tuberculosis. Our findings suggest that this species has
antituberculosis activity against sensitive and resistant strains
of Mycobacterium tuberculosis. Some phytotoxic lignans have
been isolated from this plant (Rimando et al., 1999) and calmod-
ulin inhibitors have been reported in the closely related species
Leucophyllum ambiguum (Rojas et al., 2003). However, there are
no previous reports on the antituberculosis activity and chemi-
cal nature of the antituberculosis compounds in Leucophyllum
frutescens. Tinctures obtained form Leucophyllum frutescens
440 G.M. Molina-Salinas et al. / Journal of Ethnopharmacology 109 (2007) 435–441
resulted inactive against Staphylococcus aureus, Escherichia
coli and Pseudomonas aeruginosa; the test was conducted by
agar disk susceptibility method; in addition, there is no mention
about the part of the plant investigated (Romero et al., 2005).
Chrysanctinia mexicana was extracted with diethyl ether
before extraction with methanol because of its high essential oil
content. The ethyl ether extract showed great activity against
both strains of Mycobacterium tuberculosis. This plant has
been reported to have antimycobacterial activity (Cantrell et al.,
1998). Phytochemical analysis isolated and identified monoter-
penes, but no biological activity was reported (Delgado and
Rios, 1991). Recently the antifungal activity of the essential oil
obtained from leaves of this plant has been reported (Cardenas-
Ortega et al., 2005).
Although Cordia boissieri is traditionally used for the treat-
ment of tuberculosis, our results do not show that it has good
antituberculosis activity. However, the methanol-based extract
is a good candidate for treatment of infections caused by resis-
tant strains of Streptococcus pneumoniae. Cordia curassavica
and Cordia monosperma have previously been reported to have
antimicrobial activity (De Souza et al., 2004; Hernandez et al.,
2003). Cordiaquinones have been identified as the antifungal
and larvicidal components of Cordia curassavica (Ioset et al.,
2000a) and phenylpropanoid derivatives make up the antimi-
crobial constituents of Cordia allidora (Ioset et al., 2000b). No
phytochemical investigation of Cordia boissieri has been con-
ducted.
4. Conclusions
The antimicrobial study of plants from northeast México
has identified various extracts with strong activity against sev-
eral pathogenic and antibiotic-resistant microorganisms that
cause respiratory diseases, including Mycobacterium tuberculo-
sis. These results show a good correlation between the reported
uses of the plants for respiratory infections in traditional Mexi-
can medicine and experimental data showing that such extracts
were active against at least one of the microorganisms tested. The
plants with the greatest potential as targets for bioassay-guided
fractionation are Leucophyllum frutescens, Cordia boissieri and
Chrysanctinia mexicana. Purification and identification of the
compounds responsible for their biological activity is now in
progress.
Acknowledgments
The authors wish to gratefully acknowledge the CONACYT-
México for support through grant 36544-N and doctoral fel-
lowships for GMMS and APL, as well as to the IMSS (FOFOI:
2005/1/I/021), U.A.N.L. (PAICYT) and CYTED (Project X.11).
The authors are grateful to traditional healers Milagro Aguirre
and Angélica Garcı́a and the biologist Humberto Sánchez Vega
for helping in collection and identification of the plants, as well
as to Marco Antonio Guzmán Lucio and M.C. Marı́a del Con-
suelo González de la Rosa for final taxonomic identification of
all the species reported here. Ivonne Carrera is acknowledged
for her technical assistance in the extraction procedure.
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