Influence of lipids and proteins amounts and pH values on the inhibitory effects of Origanum vulgare L. essential oil against Escherichia coli and Salmonella Typhimurium

Origanum vulgare L. (OVEO) essential oil has been considered a candidate antimicrobial for use in food conservation systems. However, studies on the influence of concomitant variations of different food components or physicochemical parameters on the antibacterial properties of OVEO are scarce. This study assessed the influence of concomitant variations in amounts of proteins - PTN (4.0, 6.0 or 8.0 g/100 mL) and lipids - LIP (3.75, 5.0 or 6.25 g/100 mL) and pH values (5.0, 5.5 or 6.0) in cultivation medium on the inhibitory effects of OVEO against Escherichia coli (EC) and Salmonella Typhimurium (ST). Lowest minimum inhibitory concentration values of OVEO against EC and ST were observed in media with the highest LIP amounts regardless the PTN amount and pH value. In absorbance based microtiter plate assay (MPA), for both EC and ST, OVEO caused the lowest Grmax values in medium containing the highest LIP and PTN amounts and lowest pH value. Highest Grmax values for EC and ST were observed in medium containing the lowest LIP and PTN amount and highest pH value. Grmax values estimated from viable counts of EC and ST in tested media with OVEO confirmed bacterial growth behavior similar to that observed in MPA. Overall, the LIP amount in media was as the most influential factor to enhance the antibacterial effects of OVEO. These results indicate that the concomitant influence of LIP and PTN amounts and pH values on the antibacterial effects of OVEO should be considered for optimizing its antimicrobial efficacy in foods.

Habituation of enterotoxigenic Staphylococcus aureus to Origanum vulgare L. essential oil does not induce direct-tolerance and cross-tolerance to salts and organic acids

Enterotoxigenic Staphylococcus aureus strains that were isolated from foods were investigated for their ability to develop direct-tolerance and cross-tolerance to sodium chloride (NaCl), potassium chloride (KCl), lactic acid (LA) and acetic acid (AA) after habituation in sublethal amounts (1/2 of the minimum inhibitory concentration - 1/2 MIC and 1/4 of the minimum inhibitory concentration - 1/4 MIC) of Origanum vulgare L. essential oil (OVEO). The habituation of S. aureus to 1/2 MIC and 1/4 MIC of OVEO did not induce direct-tolerance or cross-tolerance in the tested strains, as assessed by modulation of MIC values. Otherwise, exposing the strains to OVEO at sublethal concentrations maintained or increased the sensitivity of the cells to the tested stressing agents because the MIC values of OVEO, NaCl, KCl, LA and AA against the cells that were previously habituated to OVEO remained the same or decreased when compared with non-habituated cells. These data indicate that OVEO does not have an inductive effect on the acquisition of direct-tolerance or cross-tolerance in the tested enterotoxigenic strains of S. aureus to antimicrobial agents that are typically used in food preservation.

Standardization of sodium metabisulfite solution concentrations and immersion time for farmed shrimp Litopenaeus vannamei / Padronização da concentração da solução de metabissulfito de sódio e do tempo de imersão para camarão cultivado Litopenaeus vannamei

Sodium metabisulfite is the main additive used in the prevention of melanosis in shrimp; however, it has currently been employed with great variation in concentration by producers. Thus, the aim of the present study was to determine the correlation between the concentration of the sodium metabisulfite solution and immersion time of the whole shrimp to obtain the concentration of sulfur dioxide (SO2) in the edible muscle of farmed shrimp (Litopenaeus vannamei) in accordance with the limit established by law. For this, solutions of sodium metabisulfite at different concentrations (1%, 2 %, 3 %, 4% and 5%) were prepared and samples of L. vannamei shrimp (100g) were immersed during 10, 20 or 30 minutes at temperature of 7°C. For all treatment assayed the concentration of SO2 was determined in the edible muscle of farmed shrimp (L. vannamei). The results show that for the conditions used in this study, the correlations were linear, with significant increase (P<0.05) in the SO2 concentration in the edible muscle of shrimps both increasing sodium metabisulfite concentration as increasing immersion times, suggesting the immersion of shrimps in a 3% solution for a time of 13 minutes in order to obtain SO2 concentration of 100ppm in its edible muscle in accordance with Brazilian legislation.

O metabissulfito de sódio é o principal aditivo usado na prevenção da melanose em camarão, porém, atualmente, é empregado com grande variação de suas concentrações pelos produtores. Assim, o objetivo deste estudo foi determinar a correlação entre a concentração da solução de metabissulfito de sódio e do tempo de imersão do camarão inteiro para obter a concentração final de dióxido de enxofre (SO2) no músculo comestível de camarão cultivado (Litopenaeus vannamei), de acordo com os limites estabelecidos pela legislação. Para isso, foram preparadas soluções de metabissulfito de sódio em diferentes concentrações (1%, 2%, 3%, 4% e 5%); e amostras de camarão L. vannamei (100g) foram imersas durante 10, 20 e 30 minutos à temperatura de 7ºC. Para todos os tratamentos, foram realizadas análises da concentração de SO2 no músculo comestível do camarão cultivado (L. vannamei). Os resultados demonstraram que, para as condições empregadas nesta pesquisa, as correlações encontradas foram lineares, ocorrendo um aumento significativo (P<0,05) nos teores de SO2 no músculo comestível do camarão, tanto com o aumento da concentração das soluções de metabissulfito de sódio, quanto com o aumento no tempo de imersão, sendo possível sugerir a imersão dos camarões em solução a 3% por um tempo de 13 minutos, de forma a se obter, em seu músculo comestível, a concentração de 100ppm de SO2, de acordo com o recomendado pela legislação brasileira.

Expression of the marA, soxS, acrB and ramA genes related to the AcrAB/TolC efflux pump in Salmonella entérica strains with and without quinolone resistance-determining regions gyrA gene mutations

Several studies have been conducted in recent years to elucidate the structure, function and significance of AcrB, MarA, SoxS and RamA in Salmonella enterica. In this study, the relative quantification of acrB, soxS, marA and ramA genes expression was evaluated in 14 strains of S. enterica, with or without accompanying mutations in the quinolone resistance-determining regions of the gyrA gene, that were exposed to ciprofloxacin during the exponential growth phase. The presence of ciprofloxacin during the log phase of bacterial growth activated the genes marA, soxS, ramA and acrB in all S. enterica strains analyzed in this study. The highest expression levels for acrB were observed in strains with gyrA mutation, and marA showed the highest expression in the strains without mutation. Considering only the strains with ciprofloxacin minimum inhibitory concentration values < 0.125 [1]g/mL (sensitive to ciprofloxacin), the most expressed gene in the strains both with and without mutations was acrB. In the strains with ciprofloxacin minimum inhibitory concentration values > 0.125 [1]g/mL (low susceptibility), with and without mutations in gyrA, the most expressed gene was marA. In this study, we observed that strains resistant to nalidixic acid may express genes associated with the efflux pump and the expression of the AcrAB-TolC pump genes seems to occur independently of mutations in gyrA.

Commercially laid eggs vs. discarded hatching eggs: contamination by Salmonella spp

Salmonella enterica is frequently associated with outbreaks of human salmonellosis, and products of avian origin, such as eggs and chicken meat, are the main vehicles of its transmission. The present study describes the occurrence of different serovars of Salmonella enterica and phagotypes of S. enterica serovar Enteritidis in eggs destined for human consumption. Four thousand eggs obtained from commercial egg laying farms and one thousand discarded hatching eggs from broiler farms, which were acquired at farmers' markets and informal shops, were analyzed. Salmonella spp. was isolated from 52.0% of the discarded hatching eggs, in which the predominant serovar was Enteritidis (84.6%), and the predominant Salmonella Enteritidis phagotype (PT) was PT7 (26.9%). Salmonella spp. was not isolated from eggs obtained from commercial egg laying farms. The antimicrobial resistance profile showed that 23.1% (n = 6) of the SE strains were resistant to nalidixic acid. The results suggest that the consumption of discarded hatching eggs represents an important source of Salmonella transmission to humans.

Plasmid-mediated quinolone resistance (PMQR) and mutations in the topoisomerase genes of Salmonella enterica strains from Brazil

The objective of this study was to identify mutations in the Quinolone Resistance Determining sources Regions (QRDR) of the gyrA, gyrB, parC, and parE genes and to determine if any of the qnr variants or the aac(6')-Ib-cr variant were present in strains of Salmonella spp. isolated in Brazil. A total of 126 Salmonella spp. strains from epidemic (n = 114) and poultry (n = 12) origin were evaluated. One hundred and twelve strains (88.8%) were resistant to nalidixic acid (NAL) and 29 (23.01%) showed a reduced susceptibility to ciprofloxacin (Cip). The mutations identified were substitutions limited to the QRDR of the gyrA gene in the codons for Serine 83, Aspartate 87 and Alanine 131. The sensitivity to NAL seems to be a good phenotypic indication of distinguishing mutated and nonmutated strains in the QRDR, however the double mutation in gyrA did not cause resistance to ciprofloxacin. The qnrA1 and qnrB19 genes were detected, respectively, in one epidemic strain of S. Enteritidis and one strain of S. Corvallis of poultry origin. Despite previous detection of qnr genes in Brazil, this is the first report of qnr gene detection in Salmonella, and also the first detection of qnrB19 gene in this country. The results alert for the continuous monitoring of quinolone resistance determinants in order to minimize the emergence and selection of Salmonella spp. strains showing reduced susceptibility or resistance to quinolones.

Obtainment and characterization of mannoproteins from brewers yeast cell wall - doi: 10.4025/actascibiolsci.v34i1.7124 / Obtenção e caracterização de manoproteínas da parede celular de leveduras de descarte em cervejaria - doi: 10.4025/actascibiolsci.v34i1.7124

The biomass of yeast after beer production is a raw-material for cell components extraction, including mannoproteins. The present study evaluated the using viability of spent brewers yeast Saccharomyces sp. for obtainment of extract containing mannoprotein. The extraction was conducted by Box-Behnken 33 incomplete design, for the variables temperature (75, 85 and 95ºC), time of extraction (5, 7 and 9h) and concentration of cell wall in suspension (10, 15 and 20%). The residual ethanol of fermentation doesnt have interference in the obtaining of extract containing mannoproteins. The highest rate of extraction was 4.08%, obtained at 95ºC, with 10% cell wall by 7h and with 15% of cell wall during 9h. The experimental validation for obtaining of the maximum predicted resulted in 4.50% of extract, confirming the model predictable capacity. The extract containing mannoprotein obtained from 10% of cell wall (95ºC, 9h) had 51.39% of proteins, with 58 and 64 kDa, and 25.89% of carbohydrates, distributed in mannose and glucose. The emulsification activity was 62.50 ± 0.88% and the emulsion stability was 96.00 ± 1.4%. These results evidence the bioemulsifier potential of the extract and the viability of using spent yeast from brewery for obtainment of compounds with industrial interesting properties.

A biomass de levedura resultante da produção de cerveja é mátéria-prima para extração de componentes celulares, incluíndo manoproteínas. O presente trabalho avaliou a possibilidade da utilização da levedura Saccharomyces sp. descartada em cervejaria, para obtenção de extrato com manoproteínas. A extração foi conduzida segundo delineamento fatorial incompleto, Box-Behnken 33, para as variáveis temperaturas (75, 85 e 95ºC), tempo de extração (5, 7 e 9h) e concentração da suspensão de parede celular (10, 15 e 20%). O etanol residual da fermentação não interfere na obtenção do extrato contendo manoproteínas. O maior índice de extração foi 4,08%, observado para temperatura de 95ºC na concentração de 10% por 7h e 15% por 9h. A validação experimental do maior índice predito resultou em 4,50% de extrato, confirmando a capacidade preditiva do modelo. A manoproteína obtida, a partir de 10% de parede celular (95ºC, 9h), apresentou 51,39% de proteínas, com 58 e 64 kDa, e 25,89% de carboidratos, distribuídos entre manose e glicose. A atividade emulsificante foi de 62,50 ± 0,88% e a estabilidade da emulsão foi de 96,00 ± 1,40%. Estes resultados evidenciam o potencial bioemulsificante do extrato e a viabilidade de utilização da levedura descartada em cervejarias para obtenção de compostos com propriedades industriais interessantes

Obtenção e caracterização de manoproteínas da parede celular de leveduras de descarte em cervejaria / Obtainment and characterization of mannoproteins from brewer's yeast cell wall

A biomassa de levedura resultante da produção de cerveja é mátéria-prima para extração de componentes celulares, incluíndo manoproteínas. O presente trabalho avaliou a possibilidade da utilização da levedura Saccharomyces sp. descartada em cervejaria, para obtenção de extrato com manoproteínas. A extração foi conduzida segundo delineamento fatorial incompleto, Box-Behnken 33, para as variáveis temperaturas (75, 85 e 95ºC), tempo de extração (5, 7 e 9h) e concentração da suspensão de parede celular (10, 15 e 20%). O etanol residual da fermentação não interfere na obtenção do extrato contendo manoproteínas. O maior índice de extração foi 4,08%, observado para temperatura de 95ºC na concentração de 10% por 7h e 15% por 9h. A validação experimental do maior índice predito resultou em 4,50% de extrato, confirmando a capacidade preditiva do modelo. A manoproteína obtida, a partir de 10% de parede celular (95ºC, 9h), apresentou 51,39% de proteínas, com 58 e 64 kDa, e 25,89% de carboidratos, distribuídos entre manose e glicose. A atividade emulsificante foi de 62,50 ± 0,88% e a estabilidade da emulsão foi de 96,00 ± 1,40%. Estes resultados evidenciam o potencial bioemulsificante do extrato e a viabilidade de utilização da levedura descartada em cervejarias para obtenção de compostos com propriedades industriais interessantes.

The biomass of yeast after beer production is a raw-material for cell components extraction, including mannoproteins. The present study evaluated the using viability of spent brewer's yeast Saccharomyces sp. for obtainment of extract containing mannoprotein. The extraction was conducted by Box-Behnken 33 incomplete design, for the variables temperature (75, 85 and 95ºC), time of extraction (5, 7 and 9h) and concentration of cell wall in suspension (10, 15 and 20%). The residual ethanol of fermentation doesn't have interference in the obtaining of extract containing mannoproteins. The highest rate of extraction was 4.08%, obtained at 95ºC, with 10% cell wall by 7h and with 15% of cell wall during 9h. The experimental validation for obtaining of the maximum predicted resulted in 4.50% of extract, confirming the model predictable capacity. The extract containing mannoprotein obtained from 10% of cell wall (95ºC, 9h) had 51.39% of proteins, with 58 and 64 kDa, and 25.89% of carbohydrates, distributed in mannose and glucose. The emulsification activity was 62.50 ± 0.88% and the emulsion stability was 96.00 ± 1.4%. These results evidence the bioemulsifier potential of the extract and the viability of using spent yeast from brewery for obtainment of compounds with industrial interesting properties.

Detection of quinolone-resistance mutations in Salmonella spp. strains of epidemic and poultry origin

Mutations into codons Aspartate-87 (62 percent) and Serine-83 (38 percent) in QRDR of gyrA were identified in 105 Salmonella strains resistant to nalidixic acid (94 epidemic and 11 of poultry origin). The results show a high incidence of mutations associated to quinolone resistance but suggest association with others mechanisms of resistance.

Protective effect of carboxymethyl-glucan (CM-G) against DNA damage in patients with advanced prostate cancer

Carboxymethyl-glucan (CM-G) is a soluble derivative from Saccharomyces cerevisiae (1 → 3)(1 → 6)-β-D-glucan. The protective efficiency of CM-G against DNA damage in cells from patients with advanced prostate cancer (PCa), and undergoing Androgen Deprivation Therapy (ADT), was evaluated. DNA damage scores were obtained by the comet assay, both before and after treatment with CM-G. The reduction in DNA damage, ranging from 18 percent to 87 percent, with an average of 59 percent, was not related to the increased number of leukocytes in peripheral blood. The results demonstrate for the first time the protective effect of CM-G against DNA damage in patients with advanced PCa. Among smokers, three presented the highest reduction in DNA damage after treatment with CM-G. There was no observable relationship between DNA damage scores before and after treatment, and age, alcoholism and radiotherapy.

Ciprofloxacin susceptibility reduction of salmonella strains isolated from outbreaks

The antimicrobial susceptibility of 212 Salmonella strains isolated from patients and foods was evaluated and 45 percent were found to be resistant to nalidixic acid. Nalidixic acid resistant strains showed a higher minimal inhibitory concentration for ciprofloxacin than sensitive strains. During the study an increase of strains with reduced susceptibility to ciprofloxacin was also observed.