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Antibiotic activity of the extract of Punica granatum Linn. over bovine strains of Staphylococcus aureus

Atividade antimicrobiana do extrato de Punica granatum Linn. em linhagens de Staphylococcus aureus de origem bovina

Abstracts

Human and veterinary medicines have not been so well succeeded in order to achieving their goals concerning the treatment of infections for long term caused by Staphylococcus aureus linked to resistance development against antibiotic agents. The antibiotic activity of the Punica granatum Linn. fresh fruit pericarp extract was evaluated by the agar diffusion method on 38 S. aureus strains, isolated from apparently healthy lactating cows in farms situated in counties of the semi-arid region of the State of Paraíba, Brazil to determine the minimum inhibitory concentration (MIC). Twenty-two of the thirty-eight strains are penicillin-resistant (PRSA). The extract of P. granatum presented potential antibiotic action over all the assayed strains, forming 10 to 36 mm diameter inhibition zones. This paper's results claim the effectiveness of the extract of P. granatum as a potential antibacterial agent on S. aureus, and display the significance of evaluating new substances with antimicrobial potential, which can contribute to alternative therapeutics for veterinary and medicine.

Punica granatum; Punicacea; Staphylococcus aureus; antibiotic activity


A medicina humana e veterinária tem falhado no tratamento de infecções em longo prazo causadas por Staphylococcus aureus associadas ao desenvolvimento de resistência a agentes antimicrobianos. A atividade antimicrobiana do extrato da casca do fruto de Punica granatum Linn. foi avaliada sobre 38 linhagens de S. aureus de origem bovina para a determinação da concentração inibitória mínima (CIM). Das 38 linhagens ensaiadas, 22 são resistentes à penicilina.(PRSA). O extrato de P. granatum apresentou potencial ação antimicrobiana sobre todas as linhagens ensaiadas formando halos de inibição variando de 10 a 36 mm de diâmetro. Os resultados deste estudo confirmam a eficácia do extrato de P. granatum como potencial agente antibacteriano sobre S. aureus e demonstram a importância de se avaliar novas substâncias com potencial antimicrobiano, o que pode contribuir para a terapêutica alternativa em Medicina Veterinária.

Punica granatum; Punicacea; Staphylococcus aureus; atividade antimicrobiana


ARTIGO

Antibiotic activity of the extract of Punica granatum Linn. over bovine strains of Staphylococcus aureus

Atividade antimicrobiana do extrato de Punica granatum Linn. em linhagens de Staphylococcus aureus de origem bovina

Maria A. R. SilvaI, Jane S. HiginoII; Jozinete V. PereiraIII; José P. Siqueira-JúniorI; Maria S. V. Pereira* * E-mail: vieirapereira@uol.com.br, Tel. +55-83-32167407 , I

IDepartamento de Biologia Molecular, Centro de Ciências Exatas e da Natureza, Universidade Federal da Paraíba, 58051-970 João Pessoa-PB, Brasil

IIDepartamento de Farmácia, Centro de Ciências da Saúde, Universidade Federal de Pernambuco, Cidade Universitária, 50740-521 Recife-PE, Brasil

IIIDepartamento de Odontologia, Centro de Ciências Biológicas e da Saúde,Universidade Estadual da Paraíba, 580100-000 Campina Grande-PB, Brasil

ABSTRACT

Human and veterinary medicines have not been so well succeeded in order to achieving their goals concerning the treatment of infections for long term caused by Staphylococcus aureus linked to resistance development against antibiotic agents. The antibiotic activity of the Punica granatum Linn. fresh fruit pericarp extract was evaluated by the agar diffusion method on 38 S. aureus strains, isolated from apparently healthy lactating cows in farms situated in counties of the semi-arid region of the State of Paraíba, Brazil to determine the minimum inhibitory concentration (MIC). Twenty-two of the thirty-eight strains are penicillin-resistant (PRSA). The extract of P. granatum presented potential antibiotic action over all the assayed strains, forming 10 to 36 mm diameter inhibition zones. This paper's results claim the effectiveness of the extract of P. granatum as a potential antibacterial agent on S. aureus, and display the significance of evaluating new substances with antimicrobial potential, which can contribute to alternative therapeutics for veterinary and medicine.

Keywords:Punica granatum, Punicacea, Staphylococcus aureus, antibiotic activity.

RESUMO

A medicina humana e veterinária tem falhado no tratamento de infecções em longo prazo causadas por Staphylococcus aureus associadas ao desenvolvimento de resistência a agentes antimicrobianos. A atividade antimicrobiana do extrato da casca do fruto de Punica granatum Linn. foi avaliada sobre 38 linhagens de S. aureus de origem bovina para a determinação da concentração inibitória mínima (CIM). Das 38 linhagens ensaiadas, 22 são resistentes à penicilina.(PRSA). O extrato de P. granatum apresentou potencial ação antimicrobiana sobre todas as linhagens ensaiadas formando halos de inibição variando de 10 a 36 mm de diâmetro. Os resultados deste estudo confirmam a eficácia do extrato de P. granatum como potencial agente antibacteriano sobre S. aureus e demonstram a importância de se avaliar novas substâncias com potencial antimicrobiano, o que pode contribuir para a terapêutica alternativa em Medicina Veterinária.

Unitermos:Punica granatum, Punicacea, Staphylococcus aureus, atividade antimicrobiana.

INTRODUCTION

Staphylococcus aureus engenders a great deal of illnesses, from superficial injuries to severe systemic infections in man and other animals (Kloos & Sheleifer, 1981). It is the greatest hospital pathogen, being described as significant etiological agent in hospital infections acquired since 1950; it is also the most usual microorganism associated to goatish and bovine mastitis (Harvey & Gilmour, 1988; Watts, 1988; Shitandi & Mwangi, 2004).

The antibiotic therapy is the most employed procedure in terms of treatment of bovine mastitis, nonetheless, the increasing concern about antibiotic residues in milk, and the appearance of resistant bacterium strains have been stimulating the search for alternative ways to decrease or eliminate such problems (Pinto et al., 2001). The abusive and indiscriminate use of antibiotic agents in both medical and veterinarian practice have a bottom-up effectiveness regarding the resistance of drugs arising and maintenance (Van Wamel et al., 1995; Gillespie & Mchugh, 1997).

Plants with therapeutic properties have a great relevance in medicine throughout the world (Minja, 1994; Almeida et al., 2001; Silva et al., 2003; Rocha et al., 2005; Bezerra et al., 2006; Funke & Melzig, 2006; Leitão et al., 2006; Lima et al., 2006; Barbosa-Filho et al., 2007; Rocha et al., 2007). Punica granatum Linn. belongs to the Punicacea family, and its fruit pomegranate - known as romã in Brazil - is great. It has a yellowish rind and dark spots, and contains seeds in its core. They are sweet and astringent (Moreira, 1978). This shrub is native of northeast India, and it is cultivated all over the world, in tropical and subtropical regions (Souza, 1991). In Northeast region of Brazil this plant have been used as gargling against infections and inflammations of the tract respiratory (Agra et al., 2007; Oliveira et al., 2007). Extracts of exocarpo of fruits have exhibited giardicidal activity (Amaral et al., 2006) and might be used as an effective antibacterial alternative agent against oral biofilm bacteria (Pereira et al., 2006).

The purpose of this survey is to determine the antibiotic activity of the Punica granatum Linn. Fresh fruit pericarp extract on bovine penicillin-resistant and -sensible S. aureus strains.

MATERIAL AND METHODS

Preparation of the extract

P. granatum Linn. raw material (pomegranate) was obtained in the Public Market of João Pessoa (Paraíba), and botanically identified at the Universidade Federal de Pernambuco. The extract was obtained from the fresh fruit pericarp of pomegranate (P. granatum) at the Laboratório de Toxicologia, Universidade Federal de Pernambuco. The fruits were separated into pericarp and mesocarp. The pericarp was the material used in the experiments. After drying in a greenhouse at 33 °C, the material was ground and the active fraction extracted. The employed extraction method was the lixiviation or percolation in continuous flow at ambient temperature. The lixiviation, a process in which there is constant renewal of the extracting solution, was used (hydroalcoholic solution 95% v/v). The procedure for the extract concentration was carried out in rotatory evaporator (Ika-Werk fashion), at a constant temperature of 45 °C. The concentration at extract fluid level 1:1 (p/v).

Bacterial strains

The 38 strains of Staphylococcus aureus assayed were isolated from apparently healthy lactating cows in farms situated in counties of the semi-arid region of the State of Paraíba, Brazil (Pereira & Siqueira-Júnior, 1995).

RESULTS AND DISCUSSION

Antibiotic activity in vitro on S. aureus

This paper's results show the effectiveness of the extract of pomegranate over all the assayed strains (Table 1). The antibacterial activity afforded an inhibition zone of 10 to 36 mm. In accordance with the assayed plant's hydroalcoholic extract concentration, a homogenous growth inhibition was presented. There was a reduction of the inhibition zones diameter as the extract concentration was reduced, as displayed in table 1. The resistance to penicillin, in bovine samples of Staphylococcus aureus is a universal problem that persists since the 1950's (Wray, 1986); multiple resistance in bovine strains is not very frequent though. The assayed strains were previously characterized as for the resistance pattern. Resistance to penicillin was the most usual, followed by resistance to cadmium, streptomycin and tetracycline - antibiotics frequently used in the treatment of bovine infections. The results demonstrated an acclimation to environment pressures (Pereira & Siqueira-Júnior, 1995).

Considering the resistance pattern for penicillin, the experimented strains were characterized as penicillin-resistant S. aureus (PRSA) and penicillin-sensible S. aureus (PSSA). In the accomplishment of a comparative study with the extract of P. granatum, however, 16 of the 38 assayed strains (42,1%) presented MIC at a 1:64 dilution; 13 samples (34,2%) were inhibited at a 1:128 dilution, and 4 samples (10,5%) were inhibited at a 1:256 dilution; 1 and 3 respectively presented MIC at 1:16 and 1:32 dilutions. Both PRSA and PSSA proved to be sensible to the extract.

The antibiotic activity of the extract of P. granatum is associated to tannin phytoconstituents and alkaloids, found in leaves, roots, stem and fruits (Nawwar et al., 1994; Souza, 1991; Machado et al., 2002).

The antibiotic activity of P. granatum was also observed by other authors on several microorganisms, including S. aureus (Anesine & Perez, 1993; Lazo, 1990; Cáceres et al., 1987). Such data is congruous with our results. Michelin et al. (2005), however, observed that the P. granatum extract displayed no antibiotic activity on oxacillin resistant S. aureus.

The attained results verify the significance of gauging alternative and economically viable means to control infections in Veterinary medicine. In this context, it can be concluded that the extract of pomegranate has antibacterial activity in vitro, on PRSA and PSSA, which suggests the use of these substances in therapeutic indications in Veterinary medicine.

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  • *
    E-mail:
    vieirapereira@uol.com.br, Tel. +55-83-32167407
  • Publication Dates

    • Publication in this collection
      01 Aug 2008
    • Date of issue
      June 2008
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