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Abstract
Background
The indiscriminate use of antibiotics, associated with the emergence of microorganisms
human resistant pathogens to major classes of antibiotics, has caused many clinical
problems in the treatment of infectious diseases. Plants used in order to medicinal
purpose for treatment, healing and/or prevention is one of the oldest forms of medicinal
practice of humanity [1]. Biomonitored studies are developed and refined, relating
popular knowledge to realization of bioassays that confirm the therapeutic efficacy
and the low toxicity, as the plants with efficacy proven for a particular biological
activity are possible raw materials for natural and/or synthetic medicines [2]. The
aim of this study was to evaluate the antibacterial, cytotoxic and antioxidant potential
in vitro of extracts of a native plant from Alagoas, the Zeyheria tuberculosa, known
as Ipê Felpudo used in Brazilian folk medicine for the treatment of cancer and skin
diseases [3].
Methods
In vitro experimental research, was realized in the Laboratory of Wound Care at Federal
University of Alagoas. Four fractions were tested in different parts (leaves, stems)
of Zeyheria tuberculosa, extracts XL1. XL2, XL3 and XS1. Antimicrobial activity was
determined by microbial sensitivity tests, the method of well diffusion, plates-holes
diffusion assay and broth microdilution method for determination of minimum inhibitory
concentration (MIC). The bacterial inhibition percentage of disk diffusion test was
calculated by dividing the mean of the sample inhibition, for hundred times by the
mean of inhibition halos for the positive control[4]. Were used eight bacterial strains,
among them Gram-positive and Gram-negative bacteria, distributed by American Type
Cell Cellection. The evaluation of the samples toxicity was obtained through toxicity
test with Brine Shrimp. The evaluation of the free radical sequester ability was performed
according to the free radical sequester activity of the synthetic 2,2-Diphenyl-1-picrylhydrazyl
(DPPH).
Results and conclusions
All tested samples showed moderately active against the strain of Sthapylococcus aureus
(ATCC 25923) using the agar diffusion method by the well technique with percentage
of inhibition >25% and <75%. The XS1 extract showed the most significant antimicrobial
activity in MIC, inhibiting the strain growth of S. aureus with 1000 until 125 μg
mL-1 concentration. Identified the absence of toxicity in all samples, as the mortality
percentage rate was ≤30% (CL50 ≥ 1000 µg mL-1). In the antioxidant test, all samples
were considered inactive with CL50> 200 mg/mL. Phytochemical studies previously performed
with Z. tuberculosa showed the presence of isolated flavonoids justifying the antimicrobial
activity found [3]. These results represent the primary indications security plant
species for performing in-vivo bioassays with perspective in the infection control.
The present work was undertaken to determine safety parameters of selected Mexican medicinal plants chosen on the basis of their frequency of medicinal use and commercial importance. The medicinal herbs included Amphipteryngium adstringens, Hintonia standleyana, Hintonia latiflora, Piper sanctum, Haemathoxylon brasiletto, Iostephane heterophylla, Valeriana procera, Arracacia tolucensis, Brickellia veronicaefolia, Scaphyglottis livida, Exostema caribaeum, Hippocratea excelsa, Ligusticum porteri, Poliomintha longiflora and Gnaphalium sp. In the acute toxicity studies in mice performed according to the Lorke procedure, Exostema caribaeum, Hippocratea excelsa, Ligusticum porteri and Poliomintha longiflora were the most toxic with LD(50) values between 1085 and 2mg/kg. The Ames test revealed that Gnaphalium sp. and Valeriana procera extracts induced mutations of S. typhimurium TA98 with or without the S9 microsomal fraction, and TA100 in the presence of the enzymatic fraction, respectively. The tincture of Valeriana procera, however, was non-mutagenic. Finally, in the Artemia salina lethality test Brickellia veronicaefolia, Arracacia tolucensis, Poliomintha longiflora and Piper sanctum caused significant mortality of the crustacean larvae with LC(50) in the range of 37-227 microg/mL.
Background Due to the indiscriminate use of antimicrobial drugs, the emergence of human pathogenic microorganisms resistant to major classes of antibiotics has been increased and has caused many clinical problems in the treatment of infectious diseases. Thus, the aim of this study was to evaluate for the first time the in vitro antimicrobial activity and brine shrimp lethality of extracts and isolated compounds from Zeyheria tuberculosa (Vell.) Bur., a species used in Brazilian folk medicine for treatment of cancer and skin diseases. Methods Using the disc diffusion method, bioautography assay and brine shrimp toxicity test (Artemia salina Leach), we studied the antimicrobial activity and lethality of extracts and isolated compounds against three microorganisms strains, including Gram-positive (Staphylococcus aureus) and Gram-negative (Pseudomonas aeruginosa) bacteria and yeasts (Candida albicans). Results In this study, the extracts inhibited S. aureus (8.0 ± 0.0 to 14.0 ± 0.0 mm) and C. albicans (15.3 ± 0.68 to 25.6 ± 0.4 mm) growth. In the brine shrimp test, only two of them showed toxic effects (LC50 29.55 to 398.05 μg/mL) and some extracts were non-toxic or showed weak lethality (LC50 705.02 to > 1000 μg/mL). From these extracts, four flavones [5,6,7,8-tetramethoxyflavone (1), 5,6,7-trimethoxyflavone (2), 4'-hydroxy-5,6,7,8-tetramethoxyflavone (3), and 4'-hydroxy-5,6,7-trimethoxyflavone (4)] were isolated through bioassay-guided fractionation and identified based on the 1D and 2D NMR spectral data. By bioautography assays, compounds 1 [S. aureus (16.0 ± 0.0 mm) and C. albicans (20.0 ± 0.0 mm)] and 3 [S. aureus (10.3 ± 0.6 mm) and C. albicans (19.7 ± 0.6 mm)] inhibited both microorganisms while 2 inhibited only S. aureus (11.7 ± 0.6 mm). Compound 4 did not restrain the growth of any tested microorganism. Conclusion Our results showed that extracts and isolated flavones from Z. tuberculosa may be particularly useful against two pathogenic microorganisms, S. aureus and C. albicans. These results may justify the popular use this species since some fractions tested had antimicrobial activity and others showed significant toxic effects on brine shrimps. However, in order to evaluate possible clinical application in therapy of infectious diseases, further studies about the safety and toxicity of isolated compounds are needed.
[1
]Instituto de Tecnologia e Pesquisa, Universidade Tiradentes, Av. Murilo Dantas, 300,
Farolândia, Aracaju, Sergipe, CEP 49032-490, Brazil
[2
]Laboratório de Pesquisa em Tratamento de Feridas, Universidade Federal de Alagoas,
Campus A. C. Simões, Av. Lourival Melo Mota, s/n, Cidade Universitária, Maceió, Alagoas,
CEP 57072-900, Brazil
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Conference name:
5th Congress of the Brazilian Biotechnology Society (SBBIOTEC)
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