Maria Cristina Dantas Vanetti

Qualidade microbiológica de leite cru refrigerado e isolamento de bactérias psicrotróficas proteolíticas

Storing refrigerated raw milk in the source of production reduces economic losses which occur due to the acidifying activity of mesophilic bacteria. Nevertheless, this process selects psychrotrophic bacteria that cause technological and economic problems to the dairy industry. Samples of individual and collective refrigerated tanks and samples from an industrial silo in a dairy industry were collected and analyzed to evaluate the microbiological quality of raw milk. Furthermore, proteolytic psychrotrophic bacteria were isolated from refrigerated raw milk and characterized according to the Gram reaction and glucose fermentation. The refrigerated raw milk from the industrial silo was not compatible with legal microbiological standards, and its microbial count was significantly higher than milk from individual and collective tanks. A significant difference of contamination by mesophiles, proteolytic and non-proteolytic psychrotrophs, and Pseudomonas was observed in samples collected from refrigeration tanks compared to samples collected from the industrial silo. The Gram-negative microbiota was isolated more often, especially the glucose non-fermentative Gram-negative bacteria.

Biofilm formation and acyl homoserine lactone production in Hafnia alvei isolated from raw milk

The objective of this study was to detect the presence of acyl homoserine lactones (AHLs), signal molecules of the quorum sensing system in biofilm formed by Hafnia alvei strains. It also evaluated the effect of synthetic quorum sensing inhibitors in biofilm formation. AHLs were assayed using well diffusion techniques, thin layer chromatography (TLC) and detection directly in biofilm with biomonitors. The extracts obtained from planktonic and sessile cell of H. alvei induced at least two of three monitor strains evaluated. The presence of AHLs with up to six carbon atoms was confirmed by TLC. Biofilm formation by H. alvei was inhibited by furanone, as demonstrated by 96-well assay of crystal violet in microtitre plates and by scanning electron microscopy. The H. alvei 071 hall mutant was deficient in biofilm formation. All these results showed that the quorum sensing system is probably involved in the regulation of biofilm formation by H. alvei.

Bovicin HC5 and nisin reduce Staphylococcus aureus adhesion to polystyrene and change the hydrophobicity profile and Gibbs free energy of adhesion

Staphylococcus aureus is an opportunistic pathogen often multidrug-resistant that not only causes a variety of human diseases, but also is able to survive on biotic and abiotic surfaces through biofilm communities. The best way to inhibit biofilm establishment is to prevent cell adhesion. In the present study, subinhibitory concentrations of the bacteriocins bovicin HC5 and nisin were tested for their capability to interfere with the adhesion of S. aureus to polystyrene. Subinhibitory dosages of the bacteriocins reduced cell adhesion and this occurred probably due to changes in the hydrophobicity of the bacterial cell and polystyrene surfaces. After treatment with bovicin HC5 and nisin, the surfaces became more hydrophilic and the free energy of adhesion (∆G(adhesion)) between bacteria and the polystyrene surface was unfavorable. The transcriptional level of selected genes was assessed by RT-qPCR approach, revealing that the bacteriocins affected the expression of some important biofilm associated genes (icaD, fnbA, and clfB) and rnaIII, which is involved in the quorum sensing mechanism. The conditioning of food-contact surfaces with bacteriocins can be an innovative and powerful strategy to prevent biofilms in the food industry. The results are relevant for food safety as they indicate that bovicin HC5 and nisin can inhibit bacterial adhesion and consequent biofilm establishment, since cell adhesion precedes biofilm formation.

Enrichment of mushrooms: An interesting strategy for the acquisition of lithium

The capability of Pleurotus ostreatus mushroom to accumulate lithium (Li) and the accessibility of this Li compared with lithium carbonate (Li(2)CO(3)), often used as psychiatric medicine, were investigated. Mushrooms were produced on a substrate-based on coffee husk, with different added concentrations of lithium chloride (LiCl). Biological efficiency (BE), the crude protein content, the concentration of Li and other elements present in mushrooms were determined. The sequential extraction and in vitro test were used to verify the accessibility and the degree of solubility of this element. Li concentration in mushrooms was directly influenced by increasing LiCl concentration in the substrate (P<0.05). The BE was not affected by different concentrations of LiCl. Li present in enriched mushrooms showed greater accessibility than in Li(2)CO(3). Therefore, P. ostreatus mushrooms, enriched with lithium can be an alternative source of Li, as well as being a food with high nutritional value.

Effect of eugenol on growth and listeriolysin o production by Listeria monocytogenes

The inhibitory effect of eugenol, a naturally occurring compound mainly present in the essential oil fraction of cloves, was studied on the growth and listeriolysin O (LLO) production by Listeria monocytogenes. Potassium efflux from cells promoted by eugenol was also determined after 24 h incubation in phosphate buffered saline. Eugenol promoted a delay on the growth of L. monocytogenes at concentrations of 100, 300 and 500 µg mL-1and above 800 µg mL-1 the effect was bactericidal. Production of LLO by L. monocytogenes in the presence of eugenol was reduced 80-100%. An accumulation of external K+ was observed above 300 µg mL-1 of eugenol which indicated that the cell membrane was affected. The results showed the effectiveness of eugenol in controlling growth and LLO production of L. monocytogenes cells.

Bovicin HC5 reduces thermal resistance of Alicyclobacillus acidoterrestris in acidic mango pulp

Aims:  To test the effect of bovicin HC5 against vegetative cells and endospores of Alicyclobacillus acidoterrestris DSMZ 2498 in synthetic media and in acidic mango pulp.

In vivo bioavailability of selenium in enriched Pleurotus ostreatus mushrooms

The in vivo bioavailability of Se was investigated in enriched Pleurotus ostreatus mushrooms. A bioavailability study was performed using 64 Wistar male rats separated in 8 groups and fed with different diets: without Se, with mushrooms without Se, with enriched mushrooms containing 0.15, 0.30 or 0.45 mg kg(-1) Se and a normal diet containing 0.15 mg kg(-1) of Se using sodium selenate. The experiment was performed in two periods: depletion (14 days) and repletion (21 days), according to the Association of Official Analytical Chemists. After five weeks, the rats were sacrificed under carbon dioxide, and blood was drawn by heart puncture. Blood plasma was separated by centrifugation. The total Se concentration in the plasma of rats fed with enriched mushrooms was higher than in rats fed with a normal diet containing sodium selenate. The plasma protein profiles were obtained using size exclusion chromatography (SEC) and UV detectors. Aliquots of effluents (0.5 mL per minute) were collected throughout in the end of the chromatographic column. However, Se was determined by graphite furnace atomic absorption spectrometry (GF AAS) only in the aliquots where proteins were detected by SEC-UV. The plasma protein profile of rats fed with different diets was similar. The highest Se concentration was observed in a peptide presenting 8 kDa. Furthermore, the higher Se concentration in this peptide was obtained for rats fed with a diet using enriched mushrooms (7 μg L(-1) Se) compared to other diets (2-5 μg L(-1) Se). These results showed that Se-enriched mushrooms can be considered as an alternative Se food source for humans, due to their high bioavailability.

Bactericidal effect of bovicin HC5 and nisin against Clostridium tyrobutyricum isolated from spoiled mango pulp.

Aims:  To test the effect of bovicin HC5 – a bacteriocin from Streptococcus bovis HC5 – against the strains of Clostridium tyrobutyricum isolated from canned spoiled mango pulp.

Effect of bovicin HC5 on growth and spore germination of Bacillus cereus and Bacillus thuringiensis isolated from spoiled mango pulp

Aims:  To use bovicin HC5 to inhibit predominant bacteria isolated from spoiled mango pulp.

Isolamento de Salmonella sp e Staphylococcus aureus no processo do abate suíno como subsídio ao sistema de Análise de Perigos e Pontos Críticos de Controle - APPCC

O presente estudo foi realizado para avaliar a presenca de Salmonella sp e o numero de Staphylococcus aureus na superficie de carcacas suinas e caracterizar os perigos microbiologicos em diferentes etapas do abate e pontos criticos de controle (PCCs), atraves da quantificacao de riscos (odds ratio). Um total de 120 esfregacos superficiais de carcaca suina foi coletado em um matadouro-frigorifico, apos o escaldamento/depilacao (ponto A), antes da evisceracao (B), apos evisceracao e serragem da carcaca (C) e apos 24 horas de refrigeracao (D). Salmonella sp foi encontrada com uma frequencia media de 11,7% (14) nas carcacas, enquanto o numero de S. aureus variou entre 1,2 e 1,5 log UFC/cm2 em 11,7% das carcacas amostradas, sem evidenciar diferenca estatistica entre os pontos A, B, C e D. Pode-se concluir que os riscos de contaminacao por Salmonella sp e S. aureus foram os mesmos nas etapas do abate de suinos consideradas neste estudo.