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 »  Abstract
 »  Introduction
 »  Materials and Me...
 »  Results
 »  Discussion
 »  References
 »  Article Tables

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 Table of Contents    
RESEARCH ARTICLE
Year : 2011  |  Volume : 43  |  Issue : 2  |  Page : 180-182
 

Lantana montevidensis Briq improves the aminoglycoside activity against multiresistant Escherichia coli and Staphylococcus aureus


1 Molecular Bioprospection Postgraduation Program, Laboratory of Natural Products Research, Regional University of Cariri, 63105 - 000, Crato, CE, Brazil
2 Vice-Reitoria de Pesquisa e Pós-Graduação, Universidade de Fortaleza, Av. Washington Soares 1321, Edson Queiroz, 60811-905, Fortaleza-CE, Brazil
3 Departamento de Química, Universidade Federal do Piauí, Campus Universitário Ministro Petrônio Portella, 64049-550, Bairro Ininga,Teresina-PI, Brazil

Date of Submission26-Mar-2010
Date of Decision07-Oct-2010
Date of Acceptance31-Dec-2010
Date of Web Publication6-Mar-2011

Correspondence Address:
José G.M. Costa
Molecular Bioprospection Postgraduation Program, Laboratory of Natural Products Research, Regional University of Cariri, 63105 - 000, Crato, CE
Brazil
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0253-7613.77359

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 » Abstract 

Objective : In this work, we report the antibacterial and modulatory activity of Lantana montevidensis Briq.
Materials and Methods : The antibacterial activities of leaf (LELm) and root (RELm) extracts alone or in association with aminoglycosides were determined by a microdilution test. Multiresistant strains of Escherichia coli (Ec 27) and Staphylococcus aureus (Sa 358) were used.
Results : The results show the inhibitory activity of LELm against E. coli (minimal inhibitory concentration [MIC] 16 μg/mL) and S. aureus (MIC 128 μg/mL). The synergistic effect of the extracts and aminoglycosides was verified too. The maximum effects were obtained with RELm with gentamicin against E. coli with MIC reduction (312 to 2 μL).
Conclusion : The data from this study are indicative of the activity antibacterial of extracts of L. montevidensis and its potential in modifying the resistance of aminoglycosides.


Keywords: Aminoglycosides, antibacterial and modulatory activities, Lantana montevidensis Briq


How to cite this article:
Sousa EO, Almeida TS, Rodrigues FF, Campos AR, Lima SG, Costa JG. Lantana montevidensis Briq improves the aminoglycoside activity against multiresistant Escherichia coli and Staphylococcus aureus. Indian J Pharmacol 2011;43:180-2

How to cite this URL:
Sousa EO, Almeida TS, Rodrigues FF, Campos AR, Lima SG, Costa JG. Lantana montevidensis Briq improves the aminoglycoside activity against multiresistant Escherichia coli and Staphylococcus aureus. Indian J Pharmacol [serial online] 2011 [cited 2019 May 24];43:180-2. Available from: http://www.ijp-online.com/text.asp?2011/43/2/180/77359



 » Introduction Top


The search for new antibacterial agents is important due to the progressively increasing resistance of clinically important pathogens to known classes of antibiotics. [1] With the increased incidence of resistance to antibiotics, natural products from plants could be an interesting alternative. [2],[3] In the past years, many plants have been evaluated not only for antimicrobial activity, but also for resistance-modifying agents. [4],[5]

Resistance occurs due to random genetic mutations in the bacterial cell that alter its sensitivity to a single drug or to chemically similar drugs through a variety of mechanisms. [6] Many bacteria are able to develop changes in their sensitivity, but Staphylococcus aureus Scientific Name Search  and  Escherichia More Details coli have been recognized for the increasing resistance to conventional antibiotics.

S. aureus has persisted as one of the most important hospital and community pathogens; apart from causing different kinds of intoxication, it is usually involved in diverse tissue and/or organ infections. [7] E. coli is one of the microorganisms that has been associated with intestinal and urinary tract infections. Pathogenic and commensal strains of E. coli have different rates of resistance and can carry different genes. [8]

Lantana is a genus of about 150 species of perennial flowering plants popularly used as antirheumatics, stimulants, antibacterials, biologic controls, and as ornamental plants. [9],[10],[11],[12] Phytochemical studies of the Lantana species led to the isolation of triterpenes, steroids, and flavonoids. [13] Leaf extracts of Lantana have shown a broad spectrum of biological activities. [14],[15],[16],[17] Lantana montevidensis Briq (Verbenaceae), a shrub native to Brazil and Uruguay, is popularly known as "chumbinho." It was introduced in many countries as an ornamental plant and considered as an invasive species in many parts of the world. The leaf infusions are used in folk medicine to treat fever, influenza, asthma, bronchitis, and many other diseases. [10],[18]

The methanolic leaf extract has shown an antiproliferative activity against tumor cells and the flavonoid rich fraction was effective against human gastric adenocarcinoma, human uterine carcinoma, and melanoma cell lines. [14] However, the antimicrobial modulatory activity of L. montevidensis extracts was not previously demonstrated and this justifies this work.

In the present study, the ethanolic leaf and root extracts of L. montevidensis from Cariri Cearense, Brazilian Northeast, were evaluated as modifiers of aminoglycoside resistance.


 » Materials and Methods Top


Plant Material

Leaves and roots of Lantana montevidensis Briq were collected in March, 2009, from the Small Aromatic and Medicinal Plants Garden of the Natural Products Research Laboratory (LPPN) at University Regional do Cariri (URCA), Crato of county, Cearα state, Brazil. A voucher specimen was sent to the Herbarium Caririense Dαrdano de Andrade Lima (HCDAL), Department of Biological Sciences (URCA), which is deposited on the registration no. 1619.

Preparation of Extracts

The LELm and RELm were prepared using the cold extraction method. [19] A total of 400 g of the fresh leaves and 714 g of the roots was placed in a flask containing cold ethanol and left for 72 h at an ambient temperature. A rotary vacuum pump extractor was used to remove the ethanol from the extracts (under reduced pressure, 80ºC). The extracts were weighed and stored.

Antibacterial Activity and Minimal Inhibitory Concentration

The antibacterial activity of the LELm and RELm was investigated employing a microdilution method, recommended by National Committee for Clinical and Laboratory Standards M7-A6. [20] In the assay were used two multiresistant strains obtained from a clinical material: E. coli (Ec 27) from sputum and S. aureus (Sa 358) from a surgical wound.

The brain heart infusion (BHI 3.8%) broth was used for the bacterial growth (24 h, 35 ± 2ºC). The inoculum was an overnight culture of each bacterial species in the BHI broth diluted in the same medium to a final concentration of approximately 1 × 10 8 CFU/mL (0.5 NTU - McFarland scale). After this, the suspension was dilluted to 1 × 10 6 CFU/mL in 10% BHI. A total of 100 μL of each dilution was distributed in 96-well plates plus essential oils, achieving 5 × 10 5 CFU/mL as the final concentration of the inoculum.

The initial solution of the LELm and RELm was performed using 10 mg of each extract dissolved in 1 mL of dimethyl sulfoxide (DMSO) to obtain an initial concentration of 10 mg/mL. From this concentration, several dilutions were made in distilled water in order to obtain a stock solution of 1024 μg/mL. Further serial dilutions were performed by the addition of the BHI broth to reach a final concentration in the range of 8-512 μg/mL).

All experiments were performed in triplicate and the microdilution trays were incubated at 35 ± 2ºC for 24 h. The antibacterial activity was detected using a colorimetric method by adding 25 μL of the resauzurin staining (0.01%) aqueous solution in each well at the end of the incubation period. The minimal inhibitory concentration (MIC) was defined as the lowest extract concentration able to inhibit the bacteria growth, as indicated by resauzurin staining (dead bacterial cells are not able to change the staining color by visual observation - blue to red).

Antibiotic Modifying Activity

In order to evaluate of the LELm and RELm as modulators of antibiotic resistance, the MICs of aminoglycosides (neomycin, kanamycin, amikacin, and gentamicin) against the analyzed strains were determined in the presence or absence of the extracts using the microdutiltion test. Subinhibitory concentrations (MIC 1/8) in 10% BHI were used.

The antibiotic solutions (5000 μg/mL) were prepared in distillated water for use on the same day. A total of 100 μL of the antibiotic solution, using serial dilutions (1:2), was added to the wells containing 10% BHI and the diluted bacterial suspension (1:10). Microplates were incubated for 24 h at room temperature and the antibacterial activity was determined as described before.


 » Results Top


The antimicrobial properties of the extracts against two bacterial strains by using a microdilution assay for in vitro susceptibility testing was investigated. It was verified for the inhibitory activity, clinically relevant, against E. coli (MIC 16 μg/mL) and S. aureus (MIC 128 μg/mL), as shown in [Table 1].
Table 1: Values of the minimal inhibitory concentration (MIC) for the leaf and root extracts of L. montevidensis

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[Table 2] shows effects of extracts on aminoglycoside activities (MIC ×1/8). Reduction in MICs for all analyzed antibiotics was observed, when the extracts were added to the culture medium. The maximum effect was seen for the activity of gentamicin on E. coli by RELm, with sevenfold reduction in the MIC (312 to 2 μg/mL).
Table 2: Minimal inhibitory concentration (MIC) values for the aminoglycoside in the absence and presence of the leaf and root extracts of L. montevidensis

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 » Discussion Top


The antibacterial assay using chloroformic and methanolic extracts of L. camara leaves and three pentacyclic triterpenes isolated from hexanic extracts of L. hispida leaves showed an effective antibacterial activity against Mycobacterium tuberculosis. [15],[17] The ethanol extract of the leaves of L. camara was reported to exhibit an activity against S. aureus, Klebsiella pneumoniae, and E. coli. [21] Many triterpenes, naftoquinones, flavonoids, alkaloids, and glycosides isolated from the Lantana species are known to posses different biological activities, including antibacterial properties. [13]

Aminoglycosides are potent bactericidal antibiotics targeting the bacterial ribosome. Several mechanisms have evolved in bacteria which confer them with antibiotic resistance. These mechanisms can chemically modify the antibiotic, render it inactive through physical removal from the cell, or modify the target site so that it is not recognized by the antibiotic. In E. coli, the main mechanisms of resistance to aminoglycosides are active drug efflux and enzymatic inactivation. [22],[23]

Many substances were claimed to be modulators of the antibiotic activity, such as phenothiazines, [24] diterpenes, [25] flavones, and phenolic derivatives. [26] However, no single drug is reported to revert the aminoglycoside resistance. [27]

Several reports indicate different antibiotic combinations assayed in vitro and used in the clinics. However, the combinations of natural products and clinically used antibiotics are less reported. The data obtained in this study are indicative of the potential antibacterial and modulatory activity of the LELm and RELm.

 
 » References Top

1.Costa JG, Rodrigues FF, Angélico EC, Pereira CK, Sousa EO, Caldas GF, et al. Composição química e avaliação da atividade antibacteriana e toxicidade do óleo essencial de Croton zehntneri (variedade estragol). Braz J Pharmacogn 2007;18:583-6.  Back to cited text no. 1
    
2.Silva JG, Souza IA, Higino JS, Siqueira-Junior JP, Pereira JV, Pereira MS. Atividade antimicrobiana do extrato de Anacardium occidentale Linn em amostras multiresistentes de Staphylococcus aureus. Braz J Pharmacogn 2007;17:572-7.   Back to cited text no. 2
    
3.Coutinho HD, Costa JG, Siqueira-Júnior JP, Lima EO. In vitro anti-staphylococcal activity of Hyptis martiusii Benth against methicillin-resistant Staphylococcus aureus-MRSA strains. Braz J Pharmacogn 2008;18:670-5.  Back to cited text no. 3
    
4.Gibbons S. Anti-staphylococcal plant natural products. Nat Prod Rep 2004;21:263-77.  Back to cited text no. 4
    
5.Gurib-Fakim A. Medicinal plants: Traditions of yesterday and drugs of tomorrow. Mol Aspects Med 2006;27:1-93.  Back to cited text no. 5
    
6.Brooks GF, Butel JS, Morse JÁ. Microbiota normal do corpo humano. In: Brooks GF, Butel JS, Morse JA, editors. Microbiologia Médica. 21 nd ed. Guanabara Koogan; 2000. p. 142-5.  Back to cited text no. 6
    
7.Verhoeff J, Beaujean D, Vlok H, Baars A, Meyler A, Van Der Werkwn C, et al. A dutch approach to methicillin-resistance Staphylococcus aureus. Eur J Clin Microbiol Infect Dis 1999;18:461-6.  Back to cited text no. 7
    
8.Rosengren LB, Waldner CL, Reid-Smith RJ. Associations between antimicrobial resistance phenotypes, antimicrobial resistance genes and virulence genes of fecal Escherichia coli from healthy grow-finish pigs. Appl Environ Microbiol 2009;75:1373-380.  Back to cited text no. 8
    
9.Dua VK, Gupta NC, Pandey AC, Sharma VP. Repellency of Lantana camara (Verbenaceae) flowers against Aedes mosquitoes. J Am Mosq Control Assoc 1996;12:406-8.  Back to cited text no. 9
    
10.Ghisalberti EL. Lantana camara L. (Verbenaceae). Fitoterapia 2000;71:467-86.  Back to cited text no. 10
    
11.Costa JG, Sousa EO, Rodrigues FFG, Lima SG, Braz-Filho R. Composição química e avaliação das atividades antibacteriana e de toxicidade dos óleos essenciais de Lantana camara L. e Lantana sp. Braz J Pharmacogn 2009;19:721-5.  Back to cited text no. 11
    
12.Sousa EO, Colares AV, Rodrigues FFG, Campos AR, Lima SG, Costa JG. Effect of Collection Time on Essential Oil Composition of Lantana camara Linn (Verbenaceae) Growing in Brazil Northeastern. Rec Nat Prod 2010;4:31-7.  Back to cited text no. 12
    
13.Kohli RK, Batish DR, Batish HP, Singh HP, Dogra KS. Status, invasiveness and environmental threats of three tropical American invasive weeds (Parthenium hysterophorus L., Ageratum conyzoides L., Lantana camara L.) in India. Biol Invasions 2006;8:1501-510.  Back to cited text no. 13
    
14.Nagão T, Abe F, Kinjo J, Okabe H. Antiproliferative Constituents in Plants. Flavones from the Leaves of Lantana montevidensis BRIQ. and Consideration of Structure-Activity Relationship. Biol Pharm Bull 2002;25:875-9.  Back to cited text no. 14
    
15.Jimínez-Arellanes A, Meckes M, Torres J, Luna-Herrera J. Antimycobacterial triterpenoids from Lantana hispida (Verbenaceae). J Ethnopharmacol 2007;111:202-5.   Back to cited text no. 15
    
16.Misra N, Sharma M, Raj K, Dangi A, Srivastava S, Misra-Bhattacharya S. Chemical constituents and antifilarial activity of Lantana camara against human lymphatic filariid Brugia malayi and rodent filariid Acanthocheilonema viteae maintained in rodent hosts. Parasitol Res 2007;100:439-48.  Back to cited text no. 16
    
17.Kirimuhuzya C, Waako P, Joloba M, Odyek Olwa. The anti-mycobacterial activity of Lantana camara a plant traditionally used to treat symptoms of tuberculosis in South-western Uganda. Afr Health Sci 2009;l9:40-5.  Back to cited text no. 17
    
18.Deena MJ, Thoppil JE. Antimicrobial activity of the essential oil of Lantana. Fitoterapia 2000;71:453-5.  Back to cited text no. 18
    
19.Matos FJ, Machado MI, Craveiro AA, Barbosa-Filho JM. Essential oil of Mentha villosa Huds. J Essent Oil Res 1999;11:41-4.  Back to cited text no. 19
    
20.National Committee for Clinical Laboratory Standards. Methods for Dilution Antimicrobial Susceptibility Tests for bacteria that grow aerobically, Approved Standard. 6 th ed. NCCLS: Wayne; 2003.   Back to cited text no. 20
    
21.Nayak BS, Raju SS, Ramsubhag A. Investigation of wound healing activity of Lantana camara L. in Sprague Dawley rats using a burn wound model. Int J Applied Res Nat Prod 2008;1:15-9.  Back to cited text no. 21
    
22.Fourmy D, Recht MI, Blanchard SC, Puglisi JD. Structure of the A-site of E.coli 16 S rRNA complexed with an aminoglicoside antibiotic. Science 1996;274:1367-77.  Back to cited text no. 22
    
23.Smith E, Williamson EM, Wareham N, Kaatz GW, Gibbons S. Antibacterials and modulators of bacterial resistance from the immature cones of Chamaecyparis lawsoniana. Phytochemistry 2007;68:210-7.  Back to cited text no. 23
    
24.Gunics G, Motohashi N, Amaral L, Farkas S, Joseph M. Interaction between antibiotics and non-conventional antibiotics on bacteria. Int J Antimicrob Agents 2000;14:239-42.  Back to cited text no. 24
    
25.Nicolson K, Evans G, O'toole PW. Potentiation of methicillin activity aganst mithicillin - resistant Staphylococcal aereus by diterpenos. FEMS Microbiol let 1996;179:233-9.  Back to cited text no. 25
    
26.Sato Y, Shibata H, Arakaki N, Higuti T. 6,7 - dihydroxyflavone dramatically intensifies the susceptibility to ß - lactam antibiotics in methicillin - resistant and sensitive Staphylococcus aureus. Antimicrob Agents Chemother 2004;48:1357-360.  Back to cited text no. 26
    
27.Jana S, Deb JK. Molecular understanding of aminoglycoside action and resistance. Appl Microbiol Biotechnol 2006;70:140-50.  Back to cited text no. 27
    



 
 
    Tables

  [Table 1], [Table 2]

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