a b s t r a c tEnterococci belong to the lactic acid bacteria (LAB) group, which are often considered to provide benefit to the host organism when consumed. However, these microorganisms have a potential as infective agents, being necessary to evaluate the presence of virulence factors and resistance to antibiotics to warrant the safe use of new strains as probiotic cultures. This study aimed to detect genes of potential virulence factors related with adhesion, aggregation, biofilm formation and resistance to vancomycin, in addition to evaluate the antibiotic susceptibility and adhesion capacity of Enterococcus durans LA18s, a strain previously isolated from Minas Frescal cheese. The PCR reactions with specific primers to detect genes of adhesion collagen protein (ace), aggregation substances (agg and asa), bopA (putative glycosyltransferase), bopB (beta-phosphoglucomutase), bopC (aldose 1-epimerase), and bopD (sugar-binding transcriptional regulator) were negative for E. durans LAB18s. In addition, the strain did not present the resistance genes vanA, vanC1 and vanC2/3, and exhibited sensibility to antibiotics commonly used in animal feed, such as erythromycin, tetracycline, vancomycin, gentamicin and penicillin. This strain also showed a strong capacity of biofilm formation and exhibited satisfactory auto-aggregative and hydrophobicity features. The results suggest that this strain can be safely used in animal feed.
The genus Enterococcus belong to the genera of bacteria that produce lactic acid and can confer health benefits to living organisms. Selenium (Se) is an essential micronutrient for humans and animals. Thirty-six Enterococcus species isolated from dairy products were screened for Se(IV) sorption capacity for use as a probiotics in animal nutrition. Several isolates grew luxuriantly and significantly removed Se(IV) from Se(IV) amended medium. Two isolates, LAB 14 and LAB 18, identified by 16S rRNA gene sequence analysis as Enterococcus faecalis (98% nucleotide sequence similarity) and Enterococcus faecium (97% nucleotide sequence similarity), respectively, were selected for further studies. The two isolates grew optimally and removed selenium at initial pH 7.0. Optimum removal of Se(IV) from the medium was recorded at 25°C. Time course studies showed that after 8h of incubation LAB 14 and LAB 18 cultures displayed the highest biomass production and Se(IV) bioremoval and most selenite in culture depleted in 24h. At initial concentrations of 10 mg L(-1) and 60 mg L(-1), E. faecium (LAB 18) removed 9.91 mg L(-1) and 59.70 mg L(-1), respectively after 24h. Similar Se(IV) bioremoval capacity was recorded with E. faecalis (LAB 14). Substantial amount of Se was detected in biomass of E. faecium (0.4599 mg g(-1) of dry weight) and E. faecalis (0.4759 mg g(-1) of dry weight). The significant uptake and transformation of Se(IV) by the Enterococcus species observed in this study suggest that they can be used to deliver dietary Se through feed augmentation with Se(IV)-enriched Enterococcus biomass.
RESUMOO efeito protetor exercido por frutas e hortaliças tem sido atribuído à presença de compostos antioxidantes. Objetivou-se, neste estudo, avaliar in vitro a capacidade antioxidante de um grupo de frutas e hortaliças, cruas e cozidas, através da diminuição da peroxidação lipídica, induzida por ferro em fígado de ratos. Foram utilizados fígados de ratos homogeneizados, que foram submetidos à oxidação pelo ferro. As frutas e hortaliças foram utilizadas como antioxidantes, a fim de combater o estresse oxidativo induzido pelo ferro. O método utilizado neste trabalho foi a Reação ao Ácido Tiobarbitúrico (TBARS), tendo como marcador para avaliar o estresse oxidativo o Malonaldeído (MDA). De acordo com os resultados obtidos, observou-se que houve uma diminuição significativa do estresse oxidativo no grupo das frutas e das hortaliças cruas e cozidas com ferro, quando o fígado foi submetido à oxidação deste micronutriente. No grupo das frutas e das hortaliças cruas e cozidas sem ferro, ocorreu redução significativa do estresse oxidativo, apenas em determinadas frutas e hortaliças. O consumo de uma dieta rica em frutas e hortaliças contribui com a defesa antioxidante do organismo, inibindo danos oxidativos em macromoléculas in vitro.Termos para indexação: Antioxidantes, estresse oxidativo, peroxidação lipídica, frutas, hortaliças. ABSTRACTThe protector effect of fruits and vegetables has been attributed to the presence of antioxidant compounds. The objective of this study was to evaluate the in vitro antioxidant activity of a group of raw and cooked fruits and vegetables, through the decrease of lipid peroxidation, induced by iron in rat livers. Homogenized liver of rats that were submitted to iron oxidation were used in this experiment. The fruits and vegetables were used as antioxidants, in order to combat the oxidative stress induced by the iron. The method used in this experiment was the thiobarbituric acid reaction (TBARS), with malondialdehyde (MDA) used as a marker to evaluate the oxidative stress. In accordance with the results, a significant reduction of oxidative stress was observed in the groups of raw fruits and vegetables and fruits and vegetables cooked with iron, when the liver was submitted to the oxidation of this micronutrient. In the groups of raw fruits and vegetables and fruits and vegetables cooked without iron, a significant reduction of the oxidative stress occurred, only in certain fruits and vegetables. The consumption of a diet rich in fruits and vegetables may contribute to the antioxidant defense of the organism, inhibiting in vitro oxidative damages in macromolecules. Index terms: INTRODUÇÃOO consumo de frutas e hortaliças tem aumentado principalmente em decorrência do seu valor nutritivo e efeitos terapêuticos. Esses alimentos contêm diferentes fitoquímicos, muitos dos quais possuem propriedades antioxidantes que podem estar relacionadas com o retardo do envelhecimento e com a prevenção de certas doenças como o câncer, acompanhado de doenças-crônicas-inflamátórias, doenças cardíacas, pu...
Lactic acid bacteria (LAB) have an important role in a great variety of fermented foods. In addition to their contribution to sensory characteristics, they enhance food preservation and can be used as probiotics. In this study, the antimicrobial and antioxidant activities of culture supernatants and cell free extracts of 16 LAB isolated from meat and dairy products were investigated. The bacterial were identified by 16S rRNA sequencing. GenBank BLAST analysis revealed that all the isolates belong to Enterococcus faecium species. Antimicrobial activity against the indicator microorganism (Listeria monocytogenes) was observed at 11 culture supernatants and 4 cell free extracts. The sensibility of culture supernatant was evaluated by proteinase K and trypsin and it was observed that activity of antimicrobial substance was completely lost after the treatment. All of the isolates showed antioxidant activity as determined by the Thiobarbituric Acid Reactive Substances (TBARS) method with both types of extracts. When the antioxidant capacity was investigated using ABTS •+ method (2,2 azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)) and DPPH method (2,2-diphenyl-1-picrylhydrazyl) it was observed that only culture supernatants showed antioxidant capacity. These bacteria could particularly help to reduce or inhibit pathogenic microorganisms as well as oxidative spoilage in foods and feed.Keywords: antimicrobial activity, antioxidant, Enterococcus, lactic acid bacteria, molecular characterization.Atividade antimicrobiana e antioxidante de espécies de Enterococcus isoladas de carne e produtos lácteos ResumoAs bactérias ácido láticas (BAL) têm um papel importante em uma grande variedade de alimentos fermentados. Em adição à sua contribuição para as características sensoriais, estes microorganismos melhoram a conservação de alimentos e podem ser utilizados como probióticos. Neste estudo, as atividades antimicrobiana e antioxidante do sobrenadante e dos extratos livres de células de 16 isolados de LAB de carne e produtos lácteos foram investigadas. Os isolados foram identificados pelo sequenciamento da região 16S do rRNA. Após a comparação das sequências obtidas com aquelas disponíveis na base de dados GenBank, observou-e que todos os isolados foram pertencentes à espécie Enterococcus faecium. A atividade antimicrobiana contra o microrganismo indicador (Listeria monocytogenes) foi observada no sobrenadante das culturas em 11 isolados, e nos extratos livres de células por 4 isolados. A sensibilidade da cultura sobrenadante foi avaliada pela proteinase K e tripsina e observou-se que a atividade da substância antimicrobiana foi completamente perdida após o tratamento com as enzimas proteolíticas. Todos os isolados apresentaram atividade antioxidante, como determinado pelo método do ácido tiobarbitúrico de substâncias reativas (TBARS) com ambos os tipos de extratos. Quando a capacidade antioxidante foi investigada usando o método do ABTS (2,2 azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)) e o método de DPPH (2,2-di...
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