Vol. 8(33), pp. 1040-1050, 3 September, 2014

DOI: 10.5897/JMPR2014.5488
Article Number: 55B8C0347239
ISSN 1996-0875 Journal of Medicinal Plant Research
Copyright © 2014
Author(s) retain the copyright of this article
http://www.academicjournals.org/JMPR

Full Length Research Paper

Activities of Guatemalan medicinal plants against

cancer cell lines and selected microbes: Evidence for
their conservation
Rex G. Cates1*, Andrew Thompson1, Holly Brabazon1, Sidney McDonald1, Michael Lawrence1,
Steven Williams1, Pablo Peniallilo1, J. Alfonso Fuentes Soria2, Luis V. Espinoza3,
José Vicente Martinez4, Dany A. Arbizú5, Ernesto Villagran6 and Fernando Ancheta6
1
College of Life Sciences, Brigham Young University (BYU), Provo, UT USA.
2
Secretaría General del Consejo Superior Universitario Centroamericano (CSUCA), Ave. Las Americas 1-03, Zona No.
14, Interior Club Los Arcos, Guatemala City, Guatemala.
3
Benson Agriculture and Food Institute, Brigham Young University (BYU), Provo, UT USA.
4
Facultad de Agronomia, Universidad de San Carlos de Guatemala (USAC), Guatemala City, Guatemala.
5
Benson Institute Guatemala, Chiquimula, Guatemala.
6
Facultad de Odontología, Area Socio-Preventiva, Universidad de San Carlos de Guatemala (USAC),
Guatemala City, Guatemala.
Received 9 June, 2014; Accepted 19 August, 2014

Medicinal plants are important components in the primary health care of villagers in Guatemala.
However, an area often overlooked is the effect of medicinal plants on oral hygiene. Acetone and
methanol extracts from 63 medicinal plant species from 31 families were bioassayed against breast,
cervical, skin and tongue cancers, and the following microorganisms: Staphylococcus aureus,
Escherichia coli, Streptococcus mutans, Lactobacillus acidophilus and Candida albicans. Half-
maximum inhibitory concentrations (IC50) and half-maximum cytotoxicity concentrations (CC50) were
determined against cancerous and non-cancerous cell lines, respectively. Minimum inhibitory
concentrations (MIC) were determined against microbes. Based on levels of inhibition by extracts, IC50
values, CC50 values, and MIC values, seven species (Bursera simaruba Sarg., Burseraceae; Eriobotrya
japonica (Thumb.) Lindl., Rosaceae; Litsea guatemalensis Mez, Lauraceae; Mirabilis jalapa L.,
Nyctaginaceae; Pithecellobium dulce (Roxb.) Benth., Fabaceae); Rubus villosus Thunb., Rosaceae;
Thevetia peruviana K. Schum., Apocynaceae) were recommended for additional investigation. With
regard to oral hygiene four species (Achillea millefolium L., Asteraceae; Crotalaria longirostrata Hook.
and Arn., Fabaceae; P. dulce; Spondias purpurea L., Anacardiaceae) may merit further fractionation and
testing against oral diseases.

Key words: Anticancer, antimicrobial, IC50, CC50, MIC, oral hygiene, Guatemala.

INTRODUCTION

Well documented is the use and value of the earth’s of the drugs now available to treat human diseases are
medicinal resources with regard to primary health care for related to natural products. For anticancer, anti-migraine,
the human population. For example, Kingston (2011) and other drugs the estimate is well over 50% (Newman
and Newman and Cragg (2007) suggest that up to 50% and Cragg, 2012; Butler, 2008; McChesney et al., 2007).
 
 Cates et al. 1041

However, Newman et al. (2008), Adams and Hawkins particular interest due to their association with dental
(2007), and Chaudhuri (2007) noted that global access to plaque, caries, and other oral cavity health issues
these types of drugs is highly variable. The result is that (Kleinberg, 2002).
traditional remedies support the health care of over 65%
of the world population (Fabricant and Farnsworth, 2001),
MATERIALS AND METHODS
and in rural communities the estimate is 75 to 90%
(Chivian and Bernstein, 2008; Fowler, 2006), depending Plant collection, tissue preparation, cell lines and microbial
on the geographical area. cultures
An additional consideration is that traditional knowledge
and the biodiversity that supports that knowledge and the Eighteen species were collected from the Museo Odontológico de
Guatemala y Jardín Botánico Maya, Guatemala City, Guatemala,
development of new drugs are being lost (Cordell and 20 species from Colección y Huerto Productivo de Plantas
Colvard, 2012; Strobel et al., 2004). These in com- Medicinales, Facultad de Agronomía, Guatemala City, and 25 from
bination with the evolution of drug resistance (Lambert et the communities of Olopa and San Juan Ermita in southeastern
al., 2011) contribute to the increased awareness to Guatemala. Aids in identifying species other than vouchers and
conserve these valuable plant resources (Siwach et al., digital pictures were the Vademecum National de Plantas
Medicinales (Cáceres, 2009), the guide to medicinal plants by
2013; Kingston, 2011). Another concern regarding the Arevalo and Dieseldorff (2005), and a species list for the Museo
primary health care of people in rural communities Odontologico de Guatemala y Jardin Botánico Maya. Voucher
worldwide is the lack of information on the role of specimens are located in the herbaria at the Centro Universitario de
medicinal plants to improve oral hygiene (Colvard et al., Oriente, Universidad de San Carlos de Guatemala, Chiquimula,
2006), For example, Kufer et al. (2005) in their study on Guatemala (CUNORI) and at Brigham Young University (BRY),
the use of medicinal plants in the Ch’orto’ area in Provo, UT. Each sample from the 63 species analyzed consisted of
tissue (Table 1) collected from three or more individuals that was
southeastern Guatemala listed about 41 plants that were mixed, then bagged, labelled, and stored at -80o C (Isotemp Basic,
used to treat gastrointestinal illnesses, 34 species used Thermo Electron Corporation, Asheville, NC USA) at BYU. Acetone
for fever and pain, 38 for women’s remedies, 25 for and methanol extracts derived from five grams of plant tissue were
respiratory illnesses, but only seven for oral health eventually dissolved in double-distilled water at a final concentration
problems. Of these seven, three were used in prevention of 8 mg/ml. The human cancer cell lines used were breast (ATCC
and all seven were used for toothaches. Rural family HTB-22, breast mammary gland adenocarcinoma; ATCC,
Manassas, VA), HeLa (ATCC CCL-2, cervix epithelial
members in southeastern Guatemala near Esquipulas adenocarcinoma; ATCC), skin (ATCC CRL-2095,epithelial
who were suffering from toothache or orofacial pain malignant melanoma; ATCC), and tongue (ATCC CRL-2095,
resorted to using nine herbals but no traditional remedies human epithelial squamous carcinoma; ATCC). Cytotoxicity was
were noted to prevent cavities or other oral cavity determined using a non-cancerous Vero cell line (ATCC CRL-1586,
diseases (Hunter and Arbona, 1995). Consequently, a epithelial kidney monkey; ATCC). Staphylococcus aureus (ATCC
6538P; Becton Dickinson Laboratories, Cockeysville, MD),
need exists to find medicinal plants that have potential to Escherichia coli (ATCC 11229; ATCC) oral isolates of
prevent and treat periodontal diseases and other oral Streptococcus mutans (ATCC 33402, ATCC), Lactobacillus
health issues. acidophilus (ATCC 11975, ATCC) and Candida albicans (ATCC
These concerns are relevant to the health care of 90028, ATCC) were used to determine the antimicrobial activity of
villagers in Guatemala and therefore formed the basis for acetone and methanol extracts. Methods for culturing cancer cell
this study. The first objective was to evaluate the in vitro lines, the non-cancerous cell line, and microbes are described by
Cates et al. (2013).
growth inhibition of acetone and methanol extracts from
63 plant species against breast, cervical, skin, and
tongue cancer cell lines and a non-cancerous line. For Sulforhodamine B assay and neutral red (NR) assay
those extracts that were inhibitory at 60% or greater IC50
and CC50 values were determined. Secondly, in vitro The sulforhodamine B assay used to determine the level of
inhibition of extracts against cancer cell lines followed Skehan et al.
growth inhibition of these extracts against (1990) and Donaldson et al. (2004) as described by Cates et al.
Staphylococcus aureus, Streptococcus mutans, (2013). Inhibition activity against cell lines was determined in
Escherichia coli, Lactobacillus acidophilus, and Candida triplicate at 200, 100, and 50 µg/ml of extract. Results in Table 2
albicans were determined. For those active at 60% or are reported only for the 200 µg/ml concentration. The NR assay
greater minimum inhibitory concentrations (MIC) were followed Putnam et al. (2002) and was used on all extracts that
showed 60% or greater inhibition in the sulforhodamine assay.
obtained. All 63 species are noted in Guatemalan health
Serial dilutions of 200, 100, 50, 25, 12.5 and 6.25 µg/ml of each
care pharmacopoeias and about half of these species are plant extract were run in triplicate against each cell line (Cates et
used for oral health care. Consequently, activity against al., 2013). Additional concentrations of extract were included in the
Streptococcus mutans, Lactobacillus acidophilus, NR assay so that more data would be available for accurate
Candida albicans and the tongue cancer cell line was of calculation of half-maximum inhibitory concentrations

*Corresponding author. E-mail: [email protected].

Author(s) agree that this article remain permanently open access under the terms of the Creative Commons Attribution
License 4.0 International License

 
1042 J. Med. Plants Res.

Table 1. Scientific names, common names, tissue collected, and use of medicinal plants.

Scientific name Family Common name Tissue extracted Medicinal use

Acacia farnesiana (L.) Willd. Leguminosae Subin Leaves Vaginal bleeding, fertility, after childbirth, cold*
Acalypha guatemalensis Pax & K. Hoffm. Euphorbiaceae Hierba de cancer Leaves Gum disease, tooth ache, cancer
Achillea millefolium L. Asteraceae Milenrama Aerial portion Fever, colds, dysenteria
Allium sativum L. Liliaceae Ajo Bulb Digestion disorders, respiratory diseases
Anethum graveolens L. Apiaceae Hinojo Leaves Diarrhea, after birth antiseptic, stomach pain
Anthemis oppositifolia Lam.# Asteraceae Ixmaramac Leaves Anesthetic
Arnica montana L.† Asteraceae Arnica Aerial portion
Asclepias curassavica L. Apocynaceae Cuajatinta Leaves Fever
Baccharis trinervis Pers. Asteraceae Corrimiento Leaves Anagelsic
Bourreria huanita (Lex.) Hemsl. Boraginaceae Esquisuchil Leaves Fever, cold
Brosimum alicastrum Sw. Moraceae Ramon (Ujuxte) Green fruit Cough, sore throat
Brugmansia candida Pers. Solanaceae Florifundia Leaves Tooth ache pain, sleep agent
Bursera simaruba (L.) Sarg. Burseraceae Palo de jiote Leaves Wounds, insect bites, stings
Casimiroa edulis La Llave & Lex. Rutaceae Matasano Roots Birthing accelerant
Cedrela odorata L. Meliaceae Cedro Bark (inner) Tooth pain, birthing accelerant
Cinnamomum zeylanicum Blume Lauraceae Canela Leaves Fever, headache, cold, diarrhea
Citrus sinensis (L.) Osbeck Rutaceae Naranja Leaves Anxiety, depression
Coffea arabica L. Rubiaceae Café Leaves Dizziness
Costus pictus D. Don† Costaceae Cana de cristo Leaves
Crotalaria longirostrata Hook & Arn. Fabaceae Chipilin Leaves Sedative, anemia, insomnia
Cupressus lusitanica Mill. Cupressaceae Cipres Needles Cough
Equisetum arvense L. Equisetaceae Oreja de coche Aerial portion Gripe
Eriobotrya japonica (Thumb.) Lindl. Rosaceae Nispero Green fruit Tooth pain, gum inflammation
Euphorbia lancifolia Schldlt. Euphorbiaceae Ixbut Leaves Lactation stimulate, impotence, cold
Fleischmannia pycnocephala (Less,) R. M. King and
Asteraceae Violeta‡ Aerial portion Respiratory problems
H. Rob.
Hibiscus sabdariffa L. Malvaceae Rosa de Jamaica Leaves Intestinal distress, chicken pox
Ixora coccinea L. Rubiaceae Coralillo Leaves Muscle relaxant
Kidney and intestinal problems, heartburn, inflamed
Jatropha curcas L. Euphorbiaceae Pinon Leaves
gums
Latana camara L. Verbenaceae Cinco negritos Leaves Female hemorrhaging, discharge
Lippia dulcis Trevir. Verbenaceae Orosus Aerial portion Bronchitis
Lippia graveolens Kunth Verbenaceae Oregano Aerial portion Pain from tooth ache, spice
Litsea guatemalensis Mez Lauraceae Laurel Leaves Gastrointestinal problems, colic, swelling
Mirabilis jalapa L. Nyctaginaceae Flor de maravilla Aerial portion Cold, influenza, diarrhea
Murraya paniculata (L.) Jack Rutaceae Limonaria Leaves Relieve tooth ache pain
Nicotiana tabacum L. Solanaceae Tabaco Leaves Tooth ache
Ocimum basilicum L. Lamiaceae Albahaca morada Aerial portion Gastrointestinal spasm, migraine headaches
Ocimum micranthum Willd. Lamiaceae Albahaca del monte Aerial portion Stomach ache
Origanum vulgare L. Lamiaceae Oregano de Castillo Aerial portion Menstruation
Passiflora lingularis Juss. Heliconiaceae Granadilla Leaves Anxiety, tooth ache pain
Persea americana Mill Lauraceae Aguacate Leaves Tooth ache, rheumatism, cough
Petiveria alliacea L. Phytolacaceae Apacin Leaves Fever, nasal congestion, gastritis, diarrhea
Pimenta dioica (L.) Merr. Myrtaceae Peinata Leaves Anesthetic, stomach pain
Piper auritum Kunth Piperaceae Santa Maria Leaves Cancer
Pithecellobium dulce (Roxb.) Benth. Fabaceae Shaguay Bark Kidney stones
Priva lappulacea (L.) Pers. Verbenaceae Mozotillo Leaves Kidney disease
Prunus persica (L.) Batsch Rosaceae Duranzo Leaves Cold, cough, eating
Punica granatum L. Lythraceae Granado Leaves Tooth ache, diarrhea
Rauvolfia tetraphylla L. Apocynaceae Chalchupa Leaves Hypertension
Rhus terebinthifolia Schlech &Cham Anacardiaceae Sal de venado Leaves Tooth ache pain, gum disease
Rosmarinus officinalis L. Lamiaceae Romero Aerial portion Colic, bronchitis, anemia
Rubus villosus Lasch. Rosaceae Sarzamora Leaves Cold, cough, influenza, diarrhea, parasites
Senna occidentalis L. Fabaceae Frijolillo Leaves Dental disease
Solanum torvum Sw. Solanaceae Chichita Leaves Bronchitis, cold, diarrhea
Solanum umbellatum Miller# Solanaceae Tabaquillo Leaves Cleaning powder for teeth, tooth ache
Spondias purpurea L. Anacardiaceae Jocote Leaves Astringent, diarrhea, dysentery
 
 Cates et al. 1043

Table 1 cont’d

Stigmaphyllon ellipticum A. Juss. Malpighiaceae Contra hierba Leaves Snake bites, tooth ache
Tagetes filifolia Lag. Asteraceae Anis de monte Leaves Stomach ache, diarrhea*
Tagetes lucida Cav. Asteraceae Pericon Leaves Abdominal and menstrual pain
Taraxacum officinale F. H. Wigg. Asteraceae Amargon Leaves Hepatic and urinary disorders
Thevetia peruviana Merr. Apocynaceae Quiebra la muela Leaves Paste applied to cavity for tooth removal
Thymus vulgaris L. Lamiaceae Tomillo Aerial portion Respiratory infections, bronchitis, cough
Tridax procumbens L. Asteraceae Hierba del toro Aerial portion Hemorrhage
Vetiver grass
Vetiveria zizanioides (L.) Nash† Poaceae Leaves
(Valeriana)‡
*
Information from Kufer et al. (2005).
#
Anthemis oppositifolia and Solanum umbellatum were not analyzed for activity against microbes due to lack of tissue.
†
Medicinal use not clearly defined at time of collection.
‡
Local villagers referred to V. zizanioides as Valeriana and F. pycnocephala as violet.

(IC50) and half-maximum cytotoxicity concentrations (CC50). The against one or more of the cancer cell lines (Table 2).
IC50 and CC50 values were obtained using dosage response curves. The acetone extracts of Persea americana Mill.
(Lauraceae) and Pithecellobium dulce (Roxb.) Benth.
Microbial inhibition assay and minimum inhibitory (Fabaceae) were active against breast cancer cells (97%
concentrations (MIC) and 73% inhibition, respectively). The methanol extract
(96%) of Bursera simaruba (L.) Sarg. (Burseraceae) and
To determine which extracts exhibited inhibition against the the acetone and methanol extracts (70 and 60%,
pathogens a microwell dilution bioassay was performed using 1000,
500, and 250 µg/ml of extract following Shrestha and St. Clair
respectively) of Litsea guatemalensis Mez (Lauraceae)
(2013). Each extract was tested in triplicate and only percent were also active against this cell line. The acetone extract
inhibition at the 1000 µg/ml concentration was reported (Table 4). (94%) from P. americana and the methanol extract (75%)
For plant extracts that were inhibitory at 60% or greater (Table 4) of Cedrela odorata L. (Meliaceae) were active against the
MICs were determined using a microwell dilution bioassay. HeLa line (Table 2). Acetone and methanol (68 and
Concentrations of 1000, 500, 250, 125, 62.5, and 31.25 µg/ml were 69%, respectively) extracts from Solanum umbellatum
tested in triplicate against the microbes. The MIC was defined as
the lowest concentration of extract at which no reduction of p-
Miller (Solanaceae) and Thevetia peruviana Merr.
iodonitro-tetrazolium violet dye (Sigma-Aldrich) was observed. (Apocynaceae) (60 and 68%, respectively) also were
MICs were not calculated for S. mutans and L. acidophilus due to active against this line. Crotolaria longirostrata Hook. and
irregular growth and clumping. Details of these two assays are Arn. (Fabaceae) produced an acetone extract that was
found in Cates et al. (2013). active against skin and tongue cell lines (62% and 61%
inhibition, respectively), and the methanol extract (62%)
Data analysis of T. peruviana was active against the skin cancer cell
line (Table 2). However, the acetone extracts from C.
Data were coded by species and fraction and statistical significance longirostrata, P. dulce and the acetone and methanol
(P ≤ 0.001) between control vs. inhibition values were determined extracts from T. peruviana showed cytotoxic effects
by ANOVA (R Core Team, 2013). Results from the 200 µg/ml
against the non-cancerous Vero cell line.
concentration used against cancer cell lines and the 1000 µg/ml
concentration used against the microbes are the only results
reported (Tables 2 and 4). This is because these concentrations
yielded the maximum number of active plant species.
Neutral red (NR) assay for inhibition and cytotoxicity
Consequently, any extract showing greater than 60% inhibition for
the acetone or methanol extracts at the 200 µg/ml level for any The methanol extract from B. simaruba and the acetone
cancer cell line, and at the 1000 µg/ml for any microbial species, extract from T. peruviana were highly inhibitory at low
was considered active and worthy of neutral red or MIC analysis. concentrations (IC50 = 75 µg/ml and 30 µg/ml,
An additional criterion was that if the inhibition level of a cancer cell
line was two to three times that of the Vero line then those extracts
respectively) against the breast and HeLa cancer cell
were considered active. lines, respectively (Table 3). They also yielded low
inhibition at high concentrations against Vero cells (CC50
> 800 µg/ml and 663 µg/ml, respectively). The acetone
RESULTS extract from L. guatemalensis, and to some extent the
acetone extract from P. americana, showed moderate
Sulphorhodamine inhibition and cytotoxicity to Vero activity against the breast and HeLa lines (IC50 = 226
cells µg/ml and 387 µg/ml, respectively), and low inhibition at
high concentrations against the Vero line (CC50 > 800
Eight (12.7%) of the 63 species analyzed showed activity µg/ml). The other species showed high IC50 and/or low

 
1044 J. Med. Plants Res.

Table 2. The effect of acetone and methanol extracts on cancer cell lines.

Percent inhibition (200 μg/ml)*

Plant species Breast HeLa Skin Tongue Vero
A M A M A M A M A M
Acacia farnesiana 0 0 0 39±4 0 0 19±2 3±1 0 0
Acalypha guatemalensis 0 0 0 0 0 0 10±5 14±7 5±1 0
Achillea millifolium 0 0 0 0 0 0 0 5±2 - -
Allium sativum 0 0 0 0 0 0 0 0 - -
Anethum graveolens 5±3 0 0 4±2 0 0 0 0 - -
Anthemis oppositifolia 0 0 - - 8±4 6±1 0 0 0 0
Arnica montana 37±1 0 0 0 0 0 0 0 0 -
Asclepias curassavica 0 0 0 0 0 0 4±2 6±3 - -
Baccharis trinervis 0 0 0 0 0 0 0 0 - -
Bourreria huanita 0 0 0 0 0 0 0 0 0 0
Brosimum alicastrum 0 0 0 0 0 0 0 0 - -
Brugmansia candida 0 0 0 0 0 0 0 0 - -
Bursera simaruba 6±2 96±2 0 0 0 29±9 0 0 - 0
Casimiroa edulis 0 0 0 9±6 0 0 0 0 - -
Cedrela odorata 0 34±6 0 75±4 0 0 0 0 - 0
Cinnamomum zeylanicum 21±7 0 0 0 0 0 0 0 - -
Citrus sinensis 0 0 0 0 0 0 0 0 - -
Coffea arabica 0 0 0 0 0 0 7±3 0 - -
Costus pictus 0 0 0 27±11 0 0 0 0 - -
Crotalaria longirostrata 23±1 0 41±5 0 62±12 0 61±8 0 49±2 -
Cupressus lusitanica 0 0 0 0 0 0 0 4±2 0 3±2
Equisetum arvense 6±1 0 15 0 0 0 0 0 0 0
Eriobotrya japonica 0 0 34±6 32±7 0 0 0 0 0 0
Euphorbia lancifolia 0 0 0 0 0 0 0 0 0 0
Fleischmannia pycnocephala 0 0 0 1 5±2 4±2 0 0 0 0
Hibiscus sabdariffa 0 0 0 0 0 0 0 0 0 -
Ixora coccinea 0 0 0 9±3 0 0 0 0 - -
Jatropha curcas 0 0 0 0 0 0 0 0 0 0
Lantana camara 0 0 0 0 24±5 20±11 0 0 0 -
Lippia dulcis 0 0 0 0 0 0 2±1 0 - -
Lippia graveolens 0 0 5±1 0 0 0 0 10±3 0 0
Litsea guatemalensis 70±6 60±1 0 11±2 0 0 0 0 0 -
Mirabilis jalapa 4±1 0 0 0 27±3 0 5±1 0 0 0
Murraya paniculata 0 0 7±1 0 0 0 0 0 0 0
Nicotiana tabacum 0 0 0 0 6±1 0 12±4 0 0 0
Ocimum basilicum 3±1 0 36±8 5±2 0 0 0 0 28±2 -
Ocimum micranthum 0 0 0 8±5 0 7±1 0 0 0 0
Origanum vulgare 0 0 0 0 0 0 14±9 0 0 -
Passiflora lingularis 0 0 32±7 15±6 12±3 5±2 6±3 0 0 0
Persea americana 97±1 9±1 94±1 49±1 15±2 0 0 0 4±1 0
Petiveria alliacea 0 0 0 0 7±2 0 0 0 3±1 0
Pimenta dioica 0 0 0 0 19±4 3±1 6±1 0 0 0
Piper auritum 0 0 0 0 0 0 0 0 - -
Pithecellobium dulce 73±7 34±1 36±1 22±5 0 0 24±1 0 54±3 4±2
Priva lappulacea 46±4 0 0 0 0 0 0 0 0 -
Prunus persica 0 0 8±2 0 0 0 0 3±1 0 0
Punica granatum 0 0 21±5 0 3±2 0 0 0 7±3 0
Rauvolfia tetraphylla 0 0 0 20±1 0 0 0 0 - -
Rhus terebinthifolia 0 0 0 0 0 0 0 0 6±1 0
 
 Cates et al. 1045

Table 2 cont’d

Rosmarinus officinalis 2±1 3±1 0 0 19±6 0 0 0 - -

Rubus villosus 0 0 0 9±4 0 0 0 0 5±1 -
Senna occidentalis 0 0 0 0 11±7 0 6±3 0 - -
Solanum torvum 9±3 0 0 0 0 0 0 0 0 -
Solanum umbellatum 0 0 68±3 69±1 4±1 0 0 0 18±4 0
Spondias purpurea 18±1 0 0 0 0 0 0 0 - -
Stigmaphyllon ellipticum 0 0 0 0 0 0 0 5±3 0 0
Tagetes filifolia 0 0 12±1 0 0 0 0 0 - -
Tagetes lucida 0 0 0 0 0 3±1 0 0 5±1 0
Taraxacum officinale 0 0 0 0 0 0 6±3 0 - -
Thevetia peruviana 30±4 34±10 60±10 68±1 51±5 62±7 39±7 42±12 62±4 59±2
Thymus vulgaris 0 0 6±3 0 0 0 0 4±1 - -
Tridax procumbens 0 0 0 0 0 0 0 0 - -
Vetiveria zizanioides 0 0 0 0 0 0 0 0 0 11±5
*All comparisons between values at 60% or greater inhibition and their controls were significantly different at P ≤ 0.001.

Table 3. Half-maximum inhibitory concentration (IC50) for cancer lines and half-maximum
cytotoxicity concentration (CC50) for the Vero cell line.

IC50 (µ/ml) CC50 (µ/ml)

Cancer cell line/plant species
A M A M
Breast
Thevetia peruviana 487 592 663 <6
Bursera simaruba - 75 - >800
Pithecellobium dulce 734 - 267 -
Ocimum micranthum >800 - >800 -
Litsea guatemalensis 226 - >800 -

HeLa
Thevetia peruviana 30 85 663 <6
Persea americana 387 667 >800 >800
Solanum umbellatum 365 315 278 354

Skin
Thevetia peruviana 800 25 663 <6
Crotalaria longirostrata 168 - 136 -

Tongue
Thevetia peruviana >800 >800 663 <6
Crotalaria longirostrata 492 - 136 -

CC50 values. (Thumb.) Lindl. (Rosaceae), Mirabilis jalapa L.

(Nyctaginaceae), P. americana, Pimenta dioica (L.) Merr.
(Myrtaceae), Priva lappulacea (L.) Pers. (Verbenaceae),
Microbial inhibition and Rubus villosus Lasch. (Rosaceae) were active
against S. aureus. Methanol extracts from B. simaruba,
Thirteen (21.3%) of the 61 species tested showed growth C. odorata, and Murraya paniculata (L.) Jack (Myrtaceae)
inhibition at 60% or greater against one or more microbes were also active against S. aureus, as were the acetone
(Table 4). Acetone extracts from Eriobotrya japonica and methanol extracts from P. dulce (Table 4). Methanol

 
1046 J. Med. Plants Res.

Table 4. The effect of acetone and methanol extracts on microbes.

% Inhibition (1000 µg/ml)*

Genus/Species S. aureus S. mutans E. coli L. acidophilus C. albicans
A M A M A M A M A M
Acacia farnesiana 5±2 38±2 8±4 57±1 16±6 32±3 0 0 0 0
Acalypha guatemalensis 22±1 0 0 0 10±5 0 0 0 0 0
Achillea millefolium 0 0 9±4 51±5 8±1 95±1 20±5 98±1 0 8±3
Allium sativum 0 0 0 24±1 4±1 0 9±5 0 0 0
Anethum graveolens 5±2 0 0 7±3 0 10±4 - - 0 0
Arnica montana 22±2 0 0 26±1 0 19±3 0 21±2 0 0
Asclepias curassavica 0 16±5 12±3 19±3 0 11±1 35±5 0 0 27±2
Baccharis trinervis 0 33±4 0 0 12±2 0 0 0 5±1 0
Bourreria huanita 0 0 0 0 0 0 0 0 0 0
Brosimum alicastrum 0 12±3 10±4 0 6±2 0 0 0 0 0
Brugmansia candida 0 0 11±3 4±1 38±1 0 0 16±7 0 0
Bursera simaruba 0 68±1 23±2 36±1 14±3 56±2 0 0 0 0
Casimiroa edulis 0 58±1 12±2 37±1 12±1 5±1 0 27±6 0 0
Cedrela odorata 33±4 84±3 0 0 0 - - - 0 0
Cinnamomum zeylanicum 0 0 0 52±1 0 40±2 - - 0 0
Citrus sinensis 22±4 35±9 - 0 0 0 12±5 24±5 - 0
Coffea arabica 0 0 0 0 29±5 9±4 22±1 19±2 0 0
Costus pictus 0 0 0 0 13±7 0 17±1 39±4 0 0
Crotolaria longirostrata 0 0 0 0 22±1 0 65±3 30±1 16±4 10±7
Cupressus lusitanica 0 0 0 0 0 0 0 0 7±1 0
Equisetum arvense 0 0 0 0 0 0 0 0 0 0
Eriobotrya japonica 62±3 15±5 - 31±4 89±1 15±6 0 0 0 0
Euphorbia lancifolia 0 0 0 0 0 0 0 0 0 0
Fleischmanni pycnocephala 0 0 0 0 0 0 0 0 0 0
Hibiscus sabdariffa 0 0 15±7 19±3 21±2 0 27±2 29±1 0 21±1
Ixora coccinea 0 0 0 0 0 0 20±8 0 0 0
Jatropha curcas 0 0 20±5 16±1 10±6 0 0 0 0 0
Lantana camara 0 0 0 0 18±5 0 0 6±2 0 0
Lippia dulcis 0 0 14±2 15±1 0 0 22±2 21±4 0 20±7
Lippia graveolens 0 0 0 0 0 0 18±1 0 0 0
Litsea guatemalensis 0 0 41±5 17±4 0 0 - - 0 0
Mirabilis jalapa 60±6 48±5 36±3 0 17±4 0 0 0 0 9±3
Murraya paniculata 0 98±1 9±3 0 15±3 0 0 0 0 0
Nicotiana tabacum 0 0 0 0 0 0 0 0 0 0
Ocimum basillicum 0 0 15±3 12±3 16±6 26±1 34±2 10 0 0
Ocimum micranthum 0 0 32±3 0 0 0 0 0 0 0
Origanum vulgare 0 0 18±4 1 0 0 0 0 0 29±3
 
 Cates et al. 1047

Table 4. Cont’d.

Passiflora lingularis 0 0 0 0 0 0 0 0 0 0
Persea americana 64±5 29±4 41±3 26±2 0 0 15±3 0 0 0
Petiveria alliacea 0 0 0 0 0 0 0 6 0 0
Pimenta dioica 60±3 19±9 18±6 0 43±1 29±5 0 0 0 0
Piper auritum 0 7±1 23±4 33±6 23±2 0 0 0 0 0
Pithecellobium dulce 90±3 85±4 0 61±7 90±1 89±2 - - 0 0
Priva lappulacea 83±1 0 28±9 13±2 0 0 59±1 0 0 0
Prunus persica 0 0 - 0 14±1 12±3 10±2 15±1 0 0
Punica granatum 44±3 29±7 28±2 23±6 12±4 0 0 0 0 0
Rauvolfia tetraphylla 0 0 0 0 0 0 0 0 0 0
Rhus terebinthifolia 28±9 17±2 24±7 22±6 36±2 18±3 0 0 0 0
Rosmarinus officinalis 23±2 0 0 0 0 17±6 0 0 - 0
Rubus villosus 78±2 16±1 0 0 45±6 38±1 0 0 0 0
Senna occidentalis 0 0 0 18±2 0 0 0 23±6 0 0
Solanum torvum 0 0 0 31±3 0 0 0 0 0 0
Spondias purpurea 0 45±4 - 98±3** 0 0 35±5 13±4 0 0
Stigmaphyllon ellipticum 0 0 39±1 20±7 0 0 18±2 0 0 0
Tagetes filifolia 0 25±1 0 0 0 0 0 0 0 0
Tagetes lucida 0 0 0 0 0 0 17±3 0 0 0
Taraxacum officinale 28±4 0 0 0 0 0 0 19±5 0 0
Thevetia peruviana 0 0 16±5 0 24±2 0 0 0 0 0
Thymus vulgaris 0 0 0 13±3 0 0 22±3 25±7 0 31±6
Tridax procumbens 0 0 0 0 0 0 0 0 0 0
Vetiveria zizanioides 0 0 0 0 0 0 0 0 0 0
*All comparisons between values at 60% or greater inhibition and their controls were significantly different at P ≤ 0.001 except for S. purpurea** which was significantly
different at P ≤ 0.03.

extracts from P. dulce and Spondias purpurea L. C. albicans (Table 4). to E. coli in the inhibition assay (Table 4).
(Anacardiaceae) were inhibitory to the growth of Extracts from E. japonica and P. dulce yielded
S. mutans; no acetone extract was active against extracts with a MIC of 1000 µg/ml; all other
S. mutans (Table 4). The acetone extract from E. Minimum inhibitory concentrations (MICs) extracts yielded MIC values >1000 μg/ml and
japonica, the methanol extract from Achillea were not considered inhibitory.
millefolium L. (Asteraceae), and the acetone and The acetone extracts of M. jalapa, P. dioica, and
methanol extracts from P. dulce were active R. villosus yielded MIC values of 250 μg/ml
against E. coli. The methanol extract of A. against S. aureus (Table 5). The methanol extract DISCUSSION
millefolium and the acetone extract of C. of B. simaruba produced an MIC of >1000 μg/ml
longirostrata were the only extracts active against against S. aureus, and a MIC of 500 μg/ml against Our study along with Kufer et al. (2005) and
L. acidophilus. No extracts were active against E. coli (Table 5) even though it was not inhibitory Comerford (1996) note a wide variety of uses for
 
1048 J. Med. Plants Res.

Table 5. Minimum inhibitory concentrations (MIC) for Guatemalan medicinal plants

that showed greater than 60% inhibition against microbes.

MIC (μg/ml)
Plant species (Extract)*
S. aureus E. coli
Achillea millefolium (M) - >1000
Bursera simaruba (M) >1000 500
Cedrela odorata (M) >1000
Eribotrya japonica (A) >1000 1000
Lantana camara (M) >1000
-
Priva lappulacea (A) >1000
Mirabalis jalapa (A) 250 -
Murraya paniculata (A) >1000 >1000
Persea americana (A) >1000 -
Pimenta dioica (A) 250 -
Pithecellobium dulce (A,M) >1000 1000
Rubus villosus (A) 250 -
Spondias purpurea (A) >1000 -
*A=acetone extract; M=methanol extract; blank space indicates no inhibition per
Table 4.

the medicinal plants selected for this study (Table 1). This by villagers did not include cancer and microbial diseases
suggests that these resources are valuable to rural (Table 1) so likely the ethnomedical use will not change.
Guatemalans and need to be conserved. Overall, 16 Even so, because these species were active against
(25.4%) of 63 species were inhibitory to one or more cancer cells and microbes further study of these species
cancer cell lines and/or one or more microbes at the 60% may yield promising results.
or greater level. Eight species were inhibitory to one or One focus was to identify medicinal plant species that
more cancer cell lines and eight were inhibitory to one or might be used to improve oral hygiene. Specific
more microbes (Tables 2 and 4). Of those active against emphasis was on plant species demonstrating activity
cancer cells, extracts from B. simaruba and L. against S. mutans and L. acidophilus both of which may
guatemalensis demonstrated significant inhibition at low contribute to cavity formation, and those active against
concentrations (IC50 75 and 226 μg/ml, respectively) the tongue cancer cell line. S. purpurea and P. dulce
against the breast cell line and showed low inhibition at demonstrated significant inhibitory activity against S.
high concentrations (CC50 >800 μg/ml) against the non- mutans (Table 4). C. longirostrata was inhibitory to the
cancerous Vero cells (Table 3). The acetone extract from tongue cancer cell line (Table 2), and this species along
T. peruviana also demonstrated significant activity with A. millefolium (and P. lappulacea was almost
against the HeLa cell line (IC50 30 μg/ml vs CC50 663 inhibitory at 59% inhibition) were active against L.
μg/ml). P. americana showed some activity against the acidophilus. These species merit further investigation as
HeLa line and with further fractionation this species might to their efficacy to prevent or treat diseases of the oral
prove effective against this line. For the eight species cavity.
that were active against one or more microbes three (M. Several species reported in this study have been repor-
jalapa, P. dioica and R. villosus) registered a MIC of 250 ted elsewhere to have activity against human diseases.
μg/ml against S. aureus. B. simaruba was inhibitory to S. For example, Johnson (1999) refers to extracts from B.
auerus (Table 4) but the MIC for the methanol extract simarubra and P. americana as being used to treat sto-
was >1000 μg/ml (Table 5). Interestingly the methanol mach cancer and tumors, respectively, and in our study
extract from B. simaruba was almost significant at 54% these species were active against breast and cervical
inhibition to E. coli (Table 4) and that level of inhibition cancer cells, respectively. Additionally, S. umbellatum is
was reflected in a moderately inhibitory MIC of 500 μg/ml an important medicinal plant in some cultures but was not
against E. coli (Table 5). Extracts from C. odorata, C. reported to have activity against cancer cell lines
longirostrata, B. simaruba, P. americana, and P. dulce (Johnson, 1999).
were inhibitory to both cancer cell lines and microbes However, in our study this species was active against
(Table 2 and 4). However, extracts from these five cervical cancer cells. In summary, data from this study
species did not demonstrate significant IC50, CC50, or MIC yielded 11 significantly active species and Cates et al.
values (Tables 3 and 5). The stated uses of these species (2013) noted seven additional active species. Miller

 
 Cates et al. 1049

(2014) found 11 other Guatemalan species that produced Conflict of interests

essential oils which were highly active against the same
set of microbes used in this study which brings the total The author(s) have not declared any conflict of interests.
to 29 active medicinal plant species. Future work is
needed to determine the pharmacological activity and
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