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Industrial Crops and Products 37 (2012) 445– 450

Contents lists available at ScienceDirect

Industrial Crops and Products

journa l h o me page: www.elsev ier .com/ locate / indcrop

ntibacterial activity of plant extracts obtained with alternative organicsolvents against food-borne pathogen bacteria

artha Mendeza, Raúl Rodrígueza,∗, Judith Ruiza, Diana Morales-Adameb, Francisco Castilloc,rancisco D. Hernández-Castilloc, Cristóbal N. Aguilara

Department of Food Research, School of Chemistry, Universidad Autónoma de Coahuila, 25000 Saltillo, Coahuila, MexicoGBS GLOBAL, S.A. de C.V., Brasilia #1000 Interior-1 CP, 25270 Saltillo, Coahuila, MexicoDepartment of Agricultural Parasitology, Universidad Autónoma Agraria Antonio Narro, Buenavista, 25315 Saltillo, Coahuila, Mexico

r t i c l e i n f o

rticle history:eceived 24 November 2010eceived in revised form 10 February 2011ccepted 20 July 2011vailable online 4 September 2011

eywords:lant-extractsrganic solvents

a b s t r a c t

The objectives of this study were: the chemical characterization of extracts from seven plants (Larreatridentata, Flourensia cernua, Lippia graveolens, Agave lechuguilla, Yucca filifera, Opuntia ficus-indica, andCarya illinoensis) which are acclimated to the Mexican semi-desert. The extracts were obtained usingSoxhlet method by water, ethanol and an infusion method using alternative organic solvents (lanolinand cocoa butter), in addition it was evaluated the antibacterial activity of semi-desert plant extractsagainst Enterobacter aerogenes, Escherichia coli, Salmonella typhi and Staphylococcus aureus. Chemical char-acterization of plant extracts showed that they are rich in secondary metabolites; cocoa butter was thenon-conventional solvent which it was possible to obtain the highest content of total tannins. It was

anolinocoa butterood-borne pathogen bacteria

not possible to identify saponins in those extracts where non conventional solvents were used. Whilein extract where non conventional solvents were used, it was only possible to detect the presence ofterpenes in creosote bush and prickly pear extracts. S. aureus was the bacterial strain that showed thehighest growth inhibition as consequence of the plant extracts. The use of semi-desert plant extractsobtained using organic solvents are a good alternative for food-borne pathogen bacteria control becauseall the bacterial growth decreased with the tested extracts.

. Introduction

Food-borne pathogen bacteria are one of the major public con-erns in both developed and developing countries and account foronsiderably high cases of illnesses attacking human and animalsTayel and El-Tras, 2010). In addition, increase of international tradef commodities and food products has raised the risk of dispersionf pathogenic bacteria from production sites to faraway places ofonsumption (Dorantes et al., 2002). The food-borne pathogen bac-eria contaminations are the big problem in livestock productionpigs, chickens, cattle, and aquatic animals) countries (Sittiwet anduangpronpitag, 2009). Thus, at the present time, it is a necessityo use the chemical preservatives to prevent the growth of foodpoiling microbes in the food industry (Natta et al., 2008). But, athe same time there has been an increasing consumer demand for

oods free or with low, if any, added synthetic preservatives becauseynthetic preservatives could be toxic to human (Agatemor, 2009).n addition, bacterial resistance to currently used antibiotics is

∗ Corresponding author. Tel.: +52 844 416 1238; fax: +52 844 4390511.E-mail addresses: raul.rodriguez@uadec.edu.mx, rrh961@hotmail.com

R. Rodríguez).

926-6690/$ – see front matter © 2011 Elsevier B.V. All rights reserved.oi:10.1016/j.indcrop.2011.07.017

© 2011 Elsevier B.V. All rights reserved.

becoming a concern to public health and the development of bacte-rial super resistant strains is resulting in currently used antibioticsfailing to end many bacterial infections (Cock, 2008). Some chemi-cal preservatives have been related to carcinogenic and teratogenicattributes as well as residual toxicity (Pundir et al., 2010).

All of the above concerns have been put pressure on the foodindustry for progressive removal of chemicals preservatives andadoption of natural alternatives to obtain its goals concerning safefood with long shelf lives (Agatemor, 2009). It is well known thatsome plant have anti-microbial activity and has been apply asthe remedy for local people even before the chemical medicinewas existed (Sittiwet and Puangpronpitag, 2009). Today, scientificresearch reveals that not only the chemical from the plant haseffect against a particular disease, but, that the antioxidant propertyof the plant extracts also gives beneficial effect to human health(Puangpronpitag and Sittiwet, 2009). In addition, the perceptionthat there is a lower incidence of adverse reactions to plant prepa-rations compared to synthetic pharmaceuticals and the reducedcost of plant preparations (Cock, 2008). Make the development of

plant extract as the anti-food borne pathogen bacteria one usefulpossibility (Sittiwet and Puangpronpitag, 2009).

The plants have been poorly explored as source of antimicrobialagents because the structure and mode of action are not known

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46 M. Mendez et al. / Industrial Cr

Sharma and Hashinga, 2004). In Mexico have been reported morehan 200 plant species with antifungal and antibacterial activ-ties, mainly against plant pathogens (Montes et al., 2000). Forxtraction of active phytochemicals against plant bacteria and fun-al pathogens, the most commonly solvents used are methanol,thanol, hexane, chloroform and diethyl ether (Gamboa et al., 2003;uerrero et al., 2007; Lira-Saldívar et al., 2007; Jasso de Rodríguezt al., 2007). The use of most of these solvents is not allowed inhe context of organic production systems. By this reason, it isecessary to search for new organic solvents which allow extractore and different polyphenols, also these new solvents must be

ecognized as safe to be used under organic production systemsCastillo et al., 2010). The plants from semi-desert areas represent

great potential as antimicrobial sources. These antimicrobial phy-ochemicals probably evolved as part of the defense mechanisms ofhe host against microbial invasion (Agatemor, 2009). These plantre healthy in most of the cases because no only on their den-ity, but their biological specialization. However, there are only feweports of using semi arid plant extracts obtained using organic sol-ents for food-borne pathogen bacteria. The objectives of this studyere: the chemical characterization of Larrea tridentata, Flourensia

ernua, Lippia graveolens, Agave lechuguilla, Yucca filifera, Opuntiacus-indica, and Carya illinoensis extracts obtained using Soxhletethod and alternative organic solvents (lanolin and cocoa butter)

nd evaluate the antibacterial activity of semi-desert plant extractsgainst Enterobacter aerogenes, Escherichia coli, Salmonella typhi andtaphylococcus aureus.

. Materials and methods

.1. Vegetal material

Leaves of creosote bush L. tridentata, tar bush F. cernua, oreganoippia graveolens, lechuguilla A. lechuguilla, and yucca Y. filifera,usks of pecan nut (C. illinoensis) and stalks of O. ficus-indica wereollected from areas nearby to Saltillo, Coahuila, Mexico duringugust and September of 2008. Each vegetal tissue was dehydratedntil 4–6% of moisture was reached and each dried sample wasrinded in a miller (Thomas Wiley) and powder was sieved at 1 mm.he fine powder obtained was stored in amber bottles or darklastic bags at room temperature until phytochemical compoundsxtraction was performed.

.2. Extraction of phytochemical compounds

The phytochemical compounds extraction was performed by solid–liquid procedure, using four solvents distilled water andthanol 70% for Soxhlet method and mineral oil emulsion with 10%anolin and cocoa butter for infusion method. For Soxhlet method,ach fine powder sample was mixed in a 1:4 (w/v) ratio withhe corresponding extracting agent. Infusion method was carriedut heating the solvent at 60 ◦C, once reached this temperature;he fine powder was added and remained under these conditionsuring 7 h. After this, extracts were filtered and stored at 5 ◦C

n containers covered with aluminum foil or amber bottles untilhytochemical compounds extracted were identified and quanti-ed.

.3. Analysis of phytochemical compounds

Rapid assays for a qualitative characterization of extracts wereade. Assays for detection of the presence of terpenes, saponins

nd tannins were employed (Sofowora, 1993; Barba, 1997; Galindot al., 1989).

d Products 37 (2012) 445– 450

2.3.1. Tannins concentrationThe concentration of hydrolysable tannins (HT) was determined

by the traditional method of Folin-Ciocalteu according to the pro-tocol reported by Makkar (1999). Condensed tannins (CT) werespetrophotometrically determined using the method reported bySwain and Hillis (1959). For condensed tannins determination, analiquot of 0.5 mL of plant extract was placed in a tube, with 3 mL ofHCl/butanol (1:9) and 0.1 mL of ferric reagent. On the other hand,it was added to a tube assay serie, sole catechin (standard) in dis-tilled water at different concentrations (0, 200, 400, 600, 800 and1000 ppm) to determine the reference curve. Tubes were pluggedtightly and were heated for 1 h in water bath at 90 ◦C. After that,tubes were leaved to cool and absorbances were read at 460 nm. Forhydrolysable tannins determination, a reference curve was done byplacing 400 �L of gallic acid at different concentrations (0, 200, 400,600, 800, 1000 ppm) in assay tubes. Gallic acid concentrations wereprepared using distilled water. Each one of the plant extract werediluted in a test tube respectively, immediately to each tube wereadded 400 �L of commercial Folin-Ciocalteu reagent, samples werevortexed and leaved for 5 min. Then 400 �L of NaCO3 (0.01 M) and2.5 mL of distilled water were added. Finally absorbances were readat 725 nm in UV/visible spectrophotometer.

2.3.2. SaponinsIn different tubes assay was placed 1 mL of each one of plant

extract and added 5 mL of deionizer water. Then the different tubeswere vortexed for 30 s and left rest for 15 min, the presence ofsaponins in each one of the different plant extracts was evaluatedaccording to the foam index (Galindo et al., 1989).

2.3.3. TerpenesIn different tubes was placed an aliquot of 0.5 mL of extract, then

it were added 2 mL of acetic anhydride. Tubes were cooled on ice.After, sulfuric acid was added carefully and evaluated; a changeof color from violet to blue indicated the presence of terpenes(Sofowora, 1993).

2.4. Food-borne pathogen bacteria

Four food-borne pathogen bacteria E. aerogenes INDRE, E. coliATCC 25922, S. typhi CDD-99, S. aureus ATCC 25923 proportionedby The Coahuila Public Health State Laboratory (Saltillo Coahuila,Mexico) were used in this study. Each bacterial strains was grownin brain-heart-infusion broth (BHIB).

2.5. Antibacterial activity evaluation

The antibacterial activity of plant extracts against the fourfood-borne pathogen bacteria was evaluated in micro-assays usingconventional sterile microplates of polystyrene. Each well of themicroplate was filled with 100 �L of sterile BHIB medium (Bioxon),50 �L of 1.5E8 bacterial cells/mL and the amount of extract depend-ing of total tannin final concentration. Two control treatments(sterile water, and BHIB medium without extract) were includedin the test. Inoculated microplates were incubated at 37 ◦C dur-ing 24 h. Bacterial growth was determined by absorbance readingat 630 nm using an ELISA microplates reader (Dynatech). Bacterialcell concentration was transformed to cells/mL using the referencecurve equation; the reference curve was performed by diluting1:100 each bacterial species, counting the number of bacterial cellsof an aliquot of this dilution was done using a Neubauer chamber,

this was done for each bacterium. Finally, cell concentrations weretransformed to percentage of bacterial inhibition. The percent ofbacterial growth inhibition (P) was estimated using as reference thecontrol treatment (without extract) as follows: P = (C−T)

C×100 , where C

M. Mendez et al. / Industrial Crops and Products 37 (2012) 445– 450 447

Table 1Analysis of variance for hydrolysable, condensed and total tannins of seven plant extracts.

F. V. GL Hydrolysable tannins Condensed tannins Total tannins

CM Pr > F CM Pr > F CM Pr > F

Repetition 2 0.01 0.9156 4.07 0.51 4.63 0.4811Plant 6 404.32 <0.0001 1560.32 <0.0001 2623.23 <0.0001Solvent 3 388.72 <0.0001 4152.90 <0.0001 2247.89 <0.0001

646.02 <0.0001 1037.48 <0.00016.14 6.24

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Table 2Total tannins content in seven plant species from the Mexican semidesert area.

Vegetal specie HT (mg/g) CT (mg/g) TT

L. Tridentata 17.3769 a 37.153 a 54.530 aF. cernua 4.7652 c 4.854 f 9.619 eL. graveolens 5.7996 b 19.090 d 24.889 cA. lechuguilla 4.2952 c 22.629 c 26.924 cY. filifera 0.7953 d 12.005 e 12.801 dO. ficus-indica 1.1573 d 29.020 b 30.177 bC. illinoensis 1.0821 d 31.980 b 33.062 b

Solvent × Plant 18 236.22 <0.0001

Error 54 0.20

Total 83

s the cell concentrations under the control treatment and T is theell concentrations under the extract treatment.

.6. Experimental design and data analysis

The experiment to determine the effect of different solventsnd vegetal species on phytochemicals extraction was establishednder a randomized complete block design with three replications.

n this case a factorial arrangement of treatments was used wherehe considered factors were: solvents with four levels (water,thanol, cacao butter and lanolin) and vegetal species with sevenevels (L. tridentata, F. cernua, L. graveolens, A. lechuguilla, Y. filifera, C.llinoensis and O. ficus-indica). Data were analyzed by PROC ANOVAsing the software SAS version 6.03. The significance value that wassed to reject the null hypothesis was p < 0.05. Where it was neededukey’s range procedure was used for mean treatments separation.

The experiment to determine the effect of different extractsn bacterial growth inhibition was established under a random-zed complete block design with three replications. In this case, aactorial arrangement of treatments was used too. The consideredactors were: bacteria specie with four levels (E. aerogenes, E. coli,.typhi, and S. aureus) and vegetal extracts with seven levels (L. tri-entata, F. cernua, L. graveolens, A. lechuguilla, Y. filifera, C. illinoensisnd O. ficus-indica). Experimental unit were 10 plate wells. Dataere analyzed by PROC ANOVA using the software SAS version 6.03.

he significance value that was used to reject the null hypothesisas p < 0.05. Where it was needed Tukey’s range procedure wassed for mean treatments separation.

. Results and discussion

.1. Hydrolysable, condensed and total tannins concentration

In Table 1 is shown that non-significant differences werebserved between repetitions, but significant differences wereppreciated for the hydrolysable tannins content among plantpecie suggesting that some plants present higher hydrolysableannins content than the other tested plant species. Also, it wasetermined that quantification of hydrolysable tannins is influ-nced by the solvent employed, so significant differences in thenteraction between plant and solvent were found, this indicateshat a specific solvent allowed higher hydrolysable tannins extrac-ion in a specific plant specie.

On the other hand, significant differences for amounts of con-ensed tannins content among plant species were observed. Thisay be due to the differences in chemical composition among the

ifferent plant species evaluated in this study. In addition, dif-erences among the type of solvent used for condensed tanninsxtraction were found; this can be explained by the solvent polar-ties. Significant differences were also observed for the solvent

nd plant interaction. This suggests that a specific combinationf solvent and plant allowed obtaining the highest yield of con-ensed tannins. For the extraction of tannins, heat has been appliedo accelerate the extraction of these compounds (Waterman and

Means with the same letter are not statistically different according to multiple rangetest of Tukey (p < 0.05).

Mole, 1994). Polyphenolic compounds (mainly tannins) in plantsare responsible for the yellow color in extracts (Galvin, 1993), thiscoincides with the results shown in this study, since most of theextracts showed a yellow color, except that from pecan husk whichshowed a dark brown color.

Total tannins (TT) represent the sum of hydrolysable and con-densed tannins. Highly significant differences were found amongsolvents and among plant species for the three response variables(HT, CT and TT), these differences may be due to the polarity of eachsolvent in the first case and the different polyphenol concentrationpresent in one specific plant specie, in the second case. It was alsodetected significant difference for the interaction solvent × plant,this suggest that the highest polyphenol concentration in one spe-cific plant specie is extracted with a specific solvent (Table 1). Itmay be mentioned that, besides the tannin content is affected bythe season of plant tissue harvesting, weather conditions, pheno-logical plant step, and plant tissue (leaves, stems, roots, etc.) (Hyderet al., 2005).

3.2. Total tannins content by plant species

Creosote bush is the plant specie that contains more HT and CTwhile Yucca, Opuntia and pecan nut husk were the samples with thelowest HT amount, statistical significant differences were observedamong these samples (Table 2). On the other hand F. cernua wasthe sample with the lowest condensed tannins content. The high-est content of tannins in creosote bush in comparison to otherplant species has been reported. Garza-Villasana (2008) reportedthat extracts of L. tridentata using methanol 70% as solvent hadhigher amounts of TH and TC than F. cernua, Turnera diffusa, Jat-ropha dioica, and C. illinoensis, while Trevino-Cueto et al. (2006)mentioned that creosote bush is a good source of condensed tan-nins and hydrolysable tannins which represent 61.12% of the totalphenolic content of this plant species.

3.3. Solvent effect on extracted tannins content

The amount of hydrolysable tannins extracted with water

and alcohol were statistically higher than those extracted withlanolin and cocoa butter (Table 3). Because, polyphenols havehydroxyl groups and carboxylic acids interact in a major way withpolar solvents. There were no significant differences between the

448 M. Mendez et al. / Industrial Crops an

Table 3Total tannins content in function of the used solvent.

Solvent TH (mg/g) TC (mg/g) TT

Water 6.5127 a 9.4854 b 15.9980 dEthanol 10.4710 a 11.1261 b 21.5974 cLanolin 1.6293 b 32.4993 a 34.1286 bCocoa butter 1.5422 b 36.4495 a 37.9916 a

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reatment means followed by the same letter are not significant different accordingo the Tukey’s range test (p ≤ 0.05).

annins amount extracted with water and alcohol, as well asetween tannins extracted with lanolin and cocoa butter. Com-letely opposite results were observed for condensed tanninsxtraction, where the highest amounts of condensed tanninsxtracted were those where lanolin or cocoa butter were used asolvent. There were no reports in the literature on the use of lanolinnd cocoa butter as extraction agents and this cannot be comparedith other results. Markom et al. (2007) reported hydrolysable tan-ins extraction by different solvents and mentioned that the bestolvents for tannins extraction were water, methanol and ethanol.owever, in the present study, the use of water as solvent is limited

n some cases where a rapid fermentation is observed per exam-le A. lechuguilla solutions. The higher amounts of polyphenolsxtracted with alternative organic solvents (lanolin and cocoa but-er) may be due to the association formed between the hydrophobicegion present in their structures, and the lipophilic region of theolyphenolic ester group, in comparison to the hydrophilic regionf the water molecule. Lanolin is a complex mixture of esters ofterols, triterpene alcohols, esters of aliphatic alcohols and mono-ydroxyesters of sterols and triterpenes and aliphatic alcoholsSchlossman and McCarthy, 1978), while cocoa butter is composedy glycerides, mainly oleo-palmitostearin, oleo-distearin, oleodi-almitin, stearo-diolein, palmito-dioleintrisaturatedtriolein andriunsaturedtriolein (Beckett, 1994). In addition, other lipophilichytochemical compounds could be extracted with these kinds oflternative solvents, mainly alkaloids, terpenes and lactone deriva-ives.

.4. Saponins content in plant extracts

Table 4 shows the presence of saponins in the different plantxtracts, according to the spume index. Galindo et al. (1989) men-ions that saponins presence can be determined by vigorouslyhaking of vegetal samples for 30 s, and if a stable spume remain

able 4ualitative saponin contents determination in plant extracts obtained using four differen

Solvent Vegetal specie

L. tridentata F. cernua L. graveolens

Water ++++ ++ ++++

Ethanol +++ ++ +++

Lanolin − − −

Cocoa butter − − −

−) absence and (+++++) highly abundant spume.

able 5ualitative terpene contents determination in plant extracts obtained using four differen

Solvent Vegetal specie

L. tridentata F. cernua L. graveolens

Water + + +

Ethanol + + +

Lanolin − − −

Cocoa butter + − −

−) absence and (+) presence of terpenes.

d Products 37 (2012) 445– 450

after 15 min, this is sign of positive response to saponins presence.This method is not recommended for a quantitative determina-tion, because spume height and density are influenced by mixingintensity (Uribe-Lamas, 1987). The plant extracts with the high-est saponin index were those of A. lechuguilla when water andethanol were used as solvents. While in those extracts where lano-lin and cocoa butter were used as solvents, saponins were absent.Pecan nut husk extracts were those that had the lowest amountsof saponins with all solvents, only when water was using as sol-vent, the sample showed a light spume. Extraction of saponinsfrom various biological materials and using multiple procedureshad been reported, however given the nature greatly polar of thesecompounds, all methods agree on the hot or cold extraction.

3.5. Terpenes content in plant extracts

In most of the different plant extract where water or ethanolwas used as solvent, it was possible to detect the presence of ter-penes, only in the case of the prickly pear aqueous extract it wasnot possible to observe presence of terpenes, but from this extractwas possible to extract terpenes with an unconventional solvent(cocoa butter). The presence of terpenes was observed in mostof the extracts from creosote bush, except for the extract wherelanolin was used as solvent (Table 5). Terpenes are the major con-stituents of plant essential oils. The bactericidal, fungicidal andantiviral properties of terpenes present in vegetal essential oils arewidely documented (Boatto et al., 1994; Ahmad et al., 1994; Singhet al., 1993; Saxena and Jain, 1990; Kumar et al., 2007).

3.6. Antibacterial activity of plant extracts

Plant extracts shown differences on antibacterial activity,pathogenic bacterial growth was most inhibited by creosote bushextracts (Fig. 1). Scalbert and Williamson (2000) mentioned thatthe antimicrobial properties of tannins may be due to: tan-nins can form complexes with enzymes and other microbialproteins thus inhibiting their functions, tannins can inhibit theelectron transport through membranes and additionally, and tan-nins can alter ions as iron and copper thus inhibiting the activityof some enzymes that may be essential for microbial life. The

pecan nut husk and prickly pear extracts promoted the lowestpathogenic bacteria inhibition percentage. León-Joublanc (2007)reported that infusion of lechuguilla in ethanol enhances bacteri-cidal activity of creosote bush against pathogenic microorganisms.

t solvents.

A. lechuguilla Y. filifera O. ficus C. illinoensis

+++++ ++++ +++ ++++++ ++++ − −− − − −− − − −

t solvents.

A. lechuguilla Y. filifera O. ficus C. illinoensis

+ + − ++ + + +− − + −− − − −

M. Mendez et al. / Industrial Crops and Products 37 (2012) 445– 450 449

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Fig. 1. Percentage of bacterial growth inhibition by seven different plant extracts.

lvarez and Gonzalez (2005) studied the effect of oregano powdernd essential oil on S. aureus, E. coli and S. typhimurium growth, find-ng better inhibitory effects with 150–200 ppm of essential oil and500 ppm of powder. On the other hand, Kim et al. (1995) studiedhe antibacterial activity of 11 essential oils and carvacrol and thy-

ol, which are major components of oregano essential oil, showinghat carvacrol, and geraniol, have a powerful bactericidal activity.t has been associated the bactericidal activity of carvacrol by itsction on membrane permeability, emphasizing that the hydroxylroup is related to its bactericidal effect (Ultee et al., 1998, 1999,002).

S. aureus and E. coli presented the highest growth inhibitionffect of the plant extracts, while Enterobacter was the bacteriumith the lowest growth inhibition. Castillo-Godina (2008) reported

hat creosote bush, damiana, dragon’s blood, pecan nut shell andar bush extracts induced growth inhibition on E. coli and E. aero-enes, up to 60% and 50% respectively. These results are similar tohose obtained in this work. As can be seen, the bacteria behavioras similar, but it was observed different percentages of bacterial

rowth inhibition, this can be attributed mainly to the chemi-al composition of these plants, which contain compounds suchs diterpens and flavones that may cause rupture of membraneUrzua et al., 2006; Cowan, 1999). The secondary metabolites aremportant in plant physiology because they contribute for resis-ance to microorganisms. It has been shown that polyphenols fromifferent plant sources have interesting properties for the con-rol of pathogens with relevance to humans (Guevara-Gonzaleznd Mendez-Sanchez, 2002). Aguilera-Carbo et al. (2008) reported

hat pomegranate polyphenols had an inhibitory effect on foodathogenic bacteria.

Fig. 2. Solvent effect on the antimicrobial activity of different plant extracts.

Fig. 3. Percentage of inhibition of bacterial growth due to the effect of plant extracts.

3.7. Solvent effect on the antimicrobial activity of extracts

Antibacterial activity varied according to the solvent used forplant extracts obtaining (Figs. 2 and 3). The ethanolic extracts weremore efficient in inhibiting the growth of the tested bacteria. Thenon-conventional solvent which induced the highest percentageof bacterial growth inhibition was lanolin. There are not reportsabout the use of extracts obtained with alternative organic solvents(lanolin and cacao butter) to inhibit food-borne pathogens, by thisreason; these results may not be compared.

4. Conclusions

The use of semi arid plant extracts obtained using organic sol-vents is a good alternative for food-borne pathogen bacteria (E.aerogenes, E. coli, S. typhi and S. aureus) control, because growth of alltested bacteria was inhibited. S. aureus showed the highest growthinhibition while E. aerogenes was that with the lowest inhibitionrate. Extracts with lanolin were those that promoted the high-est food-borne bacterial growth inhibition. Creosote bush extractswere those that had the highest polyphenols content and promotedthe highest bacterial growth inhibition.

Acknowledgements

This investigation was supported by a collaborative fundinggrant to GBS GLOBAL S.A. de C.V., INNOVAPYME-2010-01-137786from the National Council of Science and Technology of Mexico(CONACYT). M.M., J.R., and F.C. want to thank to CONACYT for theirscholarships to carry out BSc. and PhD. studies.

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