Loading...
7 results
Search Results
Now showing 1 - 7 of 7
- Evaluation of a bacteriocin-producing strain of pediococcus acidilactici as a biopreservative for "alheira", a fermented meat sausagePublication . Albano, Helena; Pinho, Catarina; Leite, Daniela; Barbosa, Joana; Silva, Joana; Carneiro, Luísa; Magalhães, Rui; Hogg, Tim; Teixeira, PaulaThis study was conducted to evaluate the ability of Pediococcus acidilactici HA-6111-2, a PA-1 bacteriocinproducing lactic acid bacterium (LAB), isolated from ‘‘Alheira” to inhibit a cocktail of Listeria innocua strains during production and shelf-life of these products. The bacteriocinogenic culture reduced the Listeria population below the detection limit (1.5log CFU/g) and had no effect on the growth of the natural LAB flora or on the pH. Pathogenic organisms were not detected in any sample. The presence of some virulence factors and antibiotic resistances of the strain to be used as a bioprotective culture were investigated. P. acidilactici HA-6111-2 did not produce any of the biogenic amines tested; no formation of biofilms was observed; more L(+)lactic acid was produced than its isomer D(-); no gelatinase, DNase or lipase activity was recorded; no structural genes for the haemolysin, enterococcal surface protein, hydrolytic compounds, aggregation protein and cell-wall adhesins were detected, no significant antibiotic resistances were found. P. acidilactici HA-6111-2 appears to have potential as a bioprotective culture during ‘‘Alheira” fermentation. Moreover, a trained panel considered the protected product to be sensorially acceptable.
- Listeria: is it a problem?Publication . Barbosa, J.; Ferreira, V.; Albano, H.; Magalhães, R.; Santos, I.; Castro, A.; Silva, J.; Almeida, G.; Teixeira, P.
- Phenotypic and genetic heterogeneity of lactic acid bacteria isolated from “Alheira”, a traditional fermented sausage produced in PortugalPublication . Albano, Helena; Reenen, Carol A. van; Todorov, Svetoslav D.; Cruz, Diana; Fraga, Luisa; Hogg, Tim; Dicks, Leon M.T.; Teixeira, PaulaThe aim of this study was to evaluate the phenotypic and genetic heterogeneity of lactic acid bacteria (LAB) isolated from ‘‘alheira”, a fermented sausage produced in Portugal. LAB were identified to genus and species level by phenotypic characteristics, using genus or speciesspecific primers and sequencing of the gene encoding 16S rRNA. Two-hundred and eighty-three isolates were grouped into 14 species. Lactobacillus plantarum was isolated from all sausages and Enterococcus faecalis from most of the samples. Low numbers of Lactobacillus paraplantarum, Lactobacillus brevis, Lactobacillus rhamnosus, Lactobacillus sakei, Lactobacillus zeae, Lactobacillus paracasei, Leuconostoc mesenteroides, Pediococcus pentosaceus, Pediococcus acidilactici, Weissella cibaria, Weissella viridescens and Enterococcus faecium were recorded. The genetic heterogeneity of L. plantarum and E. faecalis strains were determined by numerical analysis of DNA banding patterns obtained by RAPD–PCR. Strains of L. plantarum and E. faecalis were different from different producers. This study forms the basis from which starter cultures could be selected for production of ‘‘Alheira”.
- Characterization of anti-listeria bacteriocins isolated from shellfish: Potential antimicrobials to control non-fermented seafoodPublication . Pinto, Ana Luísa; Fernandes, Melissa; Pinto, Cristina; Albano, Helena; Castilho, Fernanda; Teixeira, Paula; Gibbs, Paul A.This work had as main objectives to characterize two bacteriocins produced by lactic acid bacteria (LAB) previously isolated from non-fermented seafood, in order to evaluate their potential as new food protective agents. The two bacteriocinogenic isolates were identified by Polymerase Chain Reaction (PCR) using genusand species-specific primers, and confirmed by 16S rDNA sequencing, as Enterococcus faecium and Pediococcus pentosaceus. The antimicrobial spectrum of each strain included several indicator microorganisms, some of them also isolated from seafood. Growth of Listeria innocua, L. monocytogenes, Staphylococcus aureus, Bacillus cereus and other LAB species were inhibited, although no inhibition of Gram-negative microorganisms was observed. Proteolytic, but not lipolytic or glycolytic enzymes, completely inactivated the antimicrobial effect of both cell-free supernatants confirming the proteinaceous nature of the inhibitors. The antimicrobial activity was maintained after treatment with NaCl, SDS, Triton X-100, Tween 20, Tween 80 and EDTA after 2 h or 5 h of exposure and both bacteriocins were stable over a wide range of pH and temperatures. Production of bacteriocin by E. faecium (bacALP7)was detected initially at exponential phase and reached a maximum activity of 25,600 AU/ml in the early stationary phase, whereas bacteriocin production by P. pentosaceus ALP57 (bacALP57) reached the maximum at exponential phase with 12,800 AU/ml. The bacteriocins did not kill L. monocytogenes ESB54 nor L. innocua 2030c however, cellular growth was reduced. The partially purified bacteriocins, bacALP7 and bacALP57, were below 6.5 kDa in size as determined by Tricine-SDS gel electrophoresis. E. faecium and P. pentosaceus contained DNA fragments corresponding in size to those recorded for enterocin B and pediocin PA-1, respectively. Sequencing of the fragments from both bacteriocins confirmed the homology. To our knowledge, for the first time two LAB producing bacteriocins similar to pediocin PA-1 and enterocin B, were isolated from non-fermented shellfish. The adaptation of the cultures to seafood matrices may be advantageous in terms of application as a biopreservation strategy for reduction of L. monocytogenes levels in seafood products.
- Antilisterial activity of lactic acid bacteria isolated from "Alheiras" (traditional Portuguese fermented sausages)Publication . Albano, Helena; Oliveira, Márcia; Aroso, Ricardo; Cubero, Núria; Hogg, Tim; Teixeira, PaulaA total of 226 lactic acid bacteria (LAB) isolated from ‘‘Alheira’’, a traditional Portuguese fermented sausage, were screened for antagonistic activity against some pathogenic microorganisms, including Listeria monocytogenes. The objective was to isolate LAB with antibacterial activity from ‘‘Alheiras’’ and to select strains that could be used in ‘‘Alheira’’ production. Isolates displaying antibacterial activity against Listeria innocua and L. monocytogenes were investigated for the nature of the antibacterial compounds active against these microorganisms. Results showed that two LAB cultures retained activity in the supernatants after neutralization and catalase treatment. These two strains were both identified as Pediococcus pentosaceus. The final aim of this work was to test the antilisterial activity of these two strains during storage of ‘‘Alheira mass’’ (sterilized), at 4 C. The growth of L. innocua population was significantly suppressed in the paste of ‘‘Alheira’’ when the samples were co-inoculated with the LAB strains, in comparison with the paste only inoculated with L. innocua or co-inoculated with a bacteriocin negative strain of Ped. pentosaceus (ca. 1 · 107 CFU/g after 28 days of incubation).
- Characterization of two bacteriocins produced by Pediococcus acidilactici isolated from "Alheira", a fermented sausage traditionally produced in PortugalPublication . Albano, Helena; Todorov, Svetoslav D.; Reenen, Carol A. van; Hogg, Tim; Dicks, Leon M.T.; Teixeira, PaulaLactic acid bacteria were isolated from “Alheira” sausages that have been sampled from different regions in Portugal. The sausages were produced according to different recipes and with traditional starter cultures. Two isolates (HA-6111-2 and HA-5692-3) from different sausages were identified as strains of Pediococcus acidilactici. Each strain produces a bacteriocin, designated as bacHA-6111-2 and bacHA-5692-3. Both bacteriocins are produced at low levels after 18 h of growth in MRS broth (3200 AU/ml against Enterococcus faecium HKLHS and 1600 AU/ml against Listeria innocua N27). BacHA-6111-2 and bacHA-5692-3 are between 3.5 kDa and 6.5 kDa in size, as determined by tricine-SDS-PAGE. Complete inactivation or significant reduction in antimicrobial activity was observed after treatment of cell-free supernatants with proteinase K, pronase and trypsin. No change in activity was recorded when treated with catalase. Both bacteriocins are sensitive to treatment with Triton X-114 and Triton X-100, but resistant to Tween 20, Tween 80, SDS, Oxbile, NaCl, urea and EDTA. The bacteriocins remained stable after 2 h at pH 6.0. A decrease in antibacterial activity was recorded after 60 min at 100 °C. After 60 min at 80 °C, 60 °C and 25 °C the antibacterial activity against L. innocua N27 decreased by 25%. Addition of bacHA-6111-2 and bacHA-5692-3 (1600 AU/ml) to a mid-log (5-h-old) culture of L. innocua N27 inhibited growth for 7 h. Addition of the bacteriocins (3200 AU/ml) to a mid-log (5-h-old) culture of E. faecium HKLHS repressed cell growth. The bacteriocins did not adhere to the surface of the producer cells. Both strains contain a 1044 bp DNA fragment corresponding in size to that recorded for pediocin PA-1. Sequencing of the fragments from both bacteriocins revealed homology to large sections of pedA (188 bp), pedB (338 bp) and pedC (524 bp) of pediocin PA-1 and the bacteriocins are considered similar to pediocin PA-1.
- Evaluation of survival patterns and cellular injury of pseudomonas aeruginosa in different bottled waters stored under various conditionsPublication . Teixeira, Paula; Cunha, Joaquim; Albano, Helena; Ramalho, Rita; Gibbs, PaulPseudomonas aeruginosa cells were inoculated into different waters and sampled after different periods of starvation in order to evaluate the influences of storage under daylight or dark conditions, the presence or absence of the autochthonous flora, the chemical composition of the water and the storage temperature, on survival Survival was investigated by plate counts on selective and nonselective agar media. Light, low temperature (4C) and presence of the autochthonous flora negatively influenced the survival of P. aeruginosa during starvation in water. Higher survival rates were observed in waters with high mineral content. During starvation, cells developed sensitivity to the selective medium demonstrating that research is needed in the development of new media, or improvement in the existing ones, for the enumeration of P. aeruginosa in water. Current selective media/methodologies for detecting P. aeruginosa in mineral waters may seriously underestimate the levels of or presence of this organism which might represent, in some cases, a hazard to the public health.