André Bégin

Antibacterial activity of selected fatty acids and essential oils against six meat spoilage organisms

The antibacterial activity of selected fatty acids and essential oils was examined against two gram-negative (Pseudomonas fluorescens and Serratia liquefaciens) and four gram-positive (Brochothrix thermosphacta, Carnobacterium piscicola, Lactobacillus curvatus, and Lactobacillus sake) bacteria involved in meat spoilage. Various amounts of each preservative were added to brain heart infusion or MRS (deMan, Rogosa and Sharpe) agars, and the minimum inhibitory concentration was determined for each organism. Essential oils were analysed by gas-liquid chromatography to determine the concentration of selected components commonly found in spices. B. thermosphacta, P. fluorescens and S. liquefaciens were not affected by fatty acids, and generally overcame the inhibitory effect of essential oils after 24 h of exposure. Among the fatty acids, lauric and palmitoleic acids exhibited the greatest inhibitory effect with minimum inhibitory concentrations of 250 to 500 micrograms/ml, while myristic, palmitic, stearic and oleic acids were completely ineffective. For essential oils, clove, cinnamon, pimento, and rosemary were found to be the most active. The 1/100 dilution of those oils inhibited at least five of the six tested organisms. A relationship was found between the inhibitory effect of essential oils and the presence of eugenol and cinnamaldehyde.

Inhibition of surface spoilage bacteria in processed meats by application of antimicrobial films prepared with chitosan

A study was undertaken to evaluate the feasibility of using antimicrobial films, designed to slowly release bacterial inhibitors, to improve the preservation of vacuum-packaged processed meats during refrigerated storage. The antimicrobial films were prepared by incorporating acetic or propionic acid into a chitosan matrix, with or without addition of lauric acid or cinnamaldehyde, and were applied onto bologna, regular cooked ham, or pastrami. At various times during storage, packages were opened and the amounts of antimicrobial agents remaining in the chitosan matrix were measured. Regardless of film composition or meat product type, propionic acid was nearly completely released from the chitosan matrix within 48 h of application, whereas release of acetic acid was more limited, with 2-22% of the acid remaining in chitosan after 168 h of storage. Addition of lauric acid, but not cinnamaldehyde, to the chitosan matrix generally reduced the release of acetic acid significantly (P < or = 0.05) and the release was more limited onto bologna than onto ham or pastrami. In addition, the efficacies of the various films for inhibiting bacterial growth were tested against indigenous lactic acid bacteria and Enterobacteriaceae, and against Lactobacillus sakei or Serratia liqueficiens, surface-inoculated onto the meat products. Whereas lactic acid bacteria were not affected by the antimicrobial films under study, the growth of Enterobacteriaceae and S. liquefaciens was delayed or completely inhibited as a result of film application. Strongest inhibition was observed on drier surfaces (bologna), onto which acid release was slower, and with films containing cinnamaldehyde, as a result of its greater antimicrobial activity under these conditions.

Antimicrobial films produced from chitosan.

Antimicrobial films were prepared by dissolving chitosan into hydrochloric, formic, acetic, lactic and citric acid solutions. Below 40 degrees C, the counter ions could be classified into two groups based on their effect on zero-shear-rate viscosity in 2% solutions of organic acids. Chloride and citrate produced solutions with much lower viscosities than formate, acetate and lactate. At higher temperatures, these differences vanished, and the activation energies of viscous flow were all similar between 40 and 60 degrees C. Films prepared from these solutions were evaluated in tension for Youngs modulus, stress and elongation at yield and break points. Films made from hydrochloric, formic and acetic acids were hard and brittle, whereas those from lactic and citric acids were soft and could be stretched. Good correlation was found between Youngs modulus and volume of the counter ion. Film properties are essentially governed by the volume of the counter ion and not by the interactions between this counter ion and the macromolecule. Results suggest that acetate has the maximum molecular volume above which the film strength decreases very rapidly.

Structure determination of the exopolysaccharide produced by Lactobacillus rhamnosus strains RW-9595M and R.

Exopolysaccharides (EPSs) were isolated and purified from Lactobacillus rhamnosus strains RW-9595M, which has been shown to possess cytokine-stimulating activity, and R grown under various fermentation conditions (carbon source, incubation temperature and duration). Identical (1)H NMR spectra were obtained in all cases. Molecular masses were determined by gel permeation chromatography. The primary structure was elucidated using chemical and spectroscopic techniques. Organic acid, monosaccharide and absolute configuration analyses gave the following composition: pyruvate, 1; D-glucose, 2; D-galactose, 1; and l-rhamnose, 4. Methylation analysis indicated the presence of three residues of 3-linked rhamnose, and one residue each of 2,3-linked rhamnose, 2-linked glucose, 3-linked glucose and 4,6-linked galactose. The EPS was submitted to periodate oxidation followed by borohydride reduction. Monosaccharide analysis of the resulting polysaccharide gave the new composition: rhamnose, 4; and glucose, 1. Methylation analysis confirmed the loss of the 2-linked glucose and 4,6-linked galactose residues. On the basis of one- and two-dimensional (1)H and (13)C NMR data, the structure of the native EPS was consistent with the following heptasaccharide repeating unit: [3Rha alpha-3Glc beta-3[Gal4,6(R)Py alpha-2]Rha alpha-3Rha alpha-3Rha alpha-2Glc alpha-](n) where Rha corresponds to rhamnose (6-deoxymannose) and Py corresponds to pyruvate acetal. Complete (1)H and (13)C assignments are reported for the native and the corresponding pyruvate-hydrolysed polysaccharide. Electrospray MS and MS/MS data are given for the oligosaccharide produced by Smith degradation.

Inhibitory Effect of Organic Acids upon Meat Spoilage Bacteria

The relative ability of acetic, benzoic, citric, lactic, propionic, and sorbic acids to inhibit the growth of six common meat spoilage bacteria ( Brochothrix thermosphacta , Carnobacterium piscicola , Lactobacillus curvatus , Lactobacillus sake , Pseudomonas fiuorescens , and Serratia liquefaciens ) was compared under otherwise optimum conditions (BHI or MRS broths; 20°C). Because of their low solubility in the growth media, benzoic and sorbic acids could only be used in low concentrations (below 0.15% [wt/vol]) and did not efficiently inhibit bacterial growth. All other acids totally inhibited growth at concentrations ranging from 0.1 % to 1% (wt/vol). On a weight basis, acetic acid was found to be the most inhibitory, followed by propionic. lactic, and citric acid, while the order of efficiency was reversed (citric> lactic> propionic> acetic) when the acid concentrations were expressed on a molar basis or when the acid effectiveness was evaluated relative to the concentration of undissociated molecules. Overall, the lactobacilli were the bacteria most resistant to the action of organic acids, followed by P. fiuorescens and S. liquefaciens , while B. thermosphacta and C. piscicola were considerably more sensitive.

Tail-end amphiphilic dimethylaminopyridinium-containing polymethacrylates for gene delivery

Amphiphilic dimethylaminopyridinium alkyl polymethacrylates (aPPs) were tested for gene complexation, cell cytotoxicity and in vitro gene expression for use as gene delivery agents. The aminopyridinium groups neutralized by bromide or octylsulfonate counterions were terminal moieties of side-chain spacers containing 8, 12 or 16 methylene units. This investigation measured the impact of the spacer length and the chemical nature of the counterion on the physicochemical properties and biological activity of the polyplexes formed by the complexation with DNA. The aPPs self-assembled with DNA by neutralizing the DNA phosphate charges through the pyridinium moieties. The degree of DNA condensation was higher for shorter spacer (n = 8, 12) and bromide-neutralized aPPs. Several aPP–DNA complexes formed well-defined nanoparticles, which were usually, but not always, positively charged. Their sizes ranged from 30 to 150 nm and in some cases had an internal lamellar structure visible by TEM. All of the aPPs were found to be much less cytotoxic than branched poly(ethyleneimine) [(PEI), 25 kDa]. The degree of cytotoxicity of the aPPs depended mildly on their spacer length and counterion: a longer spacer (n = 16) decreased the cell viability more than shorter spacers and, at the highest aPP concentrations tested, bromide counterions more than octylsulfonate counterions. The transfection efficiency also depended on the spacer length and counterion type. Polyplexes obtained from the bromide-neutralized aPPs with the n = 12 spacer at an aPP/DNA weight ratio of 2.5, for which negatively charged nanoparticles were formed, were found to be as efficient as PEI-based polyplexes. Interestingly, this demonstrates that endosomolytic fragments and positively charged polyplex surfaces are not required for efficient gene expression.