A new and efficient method to obtain benzalkonium chloride adapted cells of Listeria monocytogenes.

A new method to obtain benzalkonium chloride (BAC) adapted L. monocytogenes cells was developed. A factorial design was used to assess the effects of the inoculum size and BAC concentration on the adaptation (measured in terms of lethal dose 50 -LD50-) of 6 strains of Listeria monocytogenes after only one exposure. The proposed method could be applied successfully in the L. monocytogenes strains with higher adaptive capacity to BAC. In those cases, a significant empirical equation was obtained showing a positive effect of the inoculum size and a positive interaction between the effects of BAC and inoculum size on the level of adaptation achieved. However, a slight negative effect of BAC, due to the biocide, was also significant. The proposed method improves the classical method based on successive stationary phase cultures in sublethal BAC concentrations because it is less time-consuming and more effective. For the laboratory strain L. monocytogenes 5873, by applying the new procedure it was possible to increase BAC-adaptation 3.69-fold in only 33 h, whereas using the classical procedure 2.61-fold of increase was reached after 5 days. Moreover, with the new method, the maximum level of adaptation was determined for all the strains reaching surprisingly almost the same concentration of BAC (mg/l) for 5 out 6 strains. Thus, a good reference for establishing the effective concentrations of biocides to ensure the maximum level of adaptation was also determined.

The use of water and ice with bactericide to prevent onboard and onshore spoilage of refrigerated megrim (Lepidorhombus whiffiagonis).

This study investigates the effectiveness of ozonated water and flake ice (combined Petfrost system) to increase the quality and stability of fresh megrim on fishing boats. The captured fish were washed, placed in plastic boxes, covered with flake ice and refrigerated at 2°C for up to 2-weeks onboard and, thereafter, for 11 days onshore. The experiments employed sterile, filtered and ozonated water at a concentration of 2ppm for washing the fish and making the flake ice. The results are compared with samples from a traditional treatment consisting of water and flake ice of marine origin. Fish were caught in four different hauls, which took 14, 12, 8 and 3 days in being landed. Subsequently, fish were stored for 1, 5, 7 and 11 days at 3°C. The different treatments were evaluated using sensory, microbiological and chemical techniques. Fish treated with ozone always showed the best quality. Megrim treated with ozone was still suitable for consumption after 14 days on board, and megrim stored for 12, 8 and 3 days on board could be stored for a further five days in the ice state once landed with an acceptable quality. In contrast, control fish were not suitable for consumption if stored for longer than three days on board.The results indicate that treatment with water and ice flakes made from sterile and ozonated water maintains the quality of fresh megrim onboard fishing boats and upon arrival onshore.

Modified method for the analysis of free fatty acids in fish.

The objective of this study was the development of a method for the quantification of free fatty acids (FFA) using less aggressive reactants against the handler and the environment than those used in the classic method of Lowry and Tinsley. The modified procedure is a variation of the Lowry and Tinsley method employing cyclohexane in place of benzene. The use of benzene is prohibited in certain work processes and laboratories, and the competent authority in each country is actively promoting research into harmless or less harmful products that could replace benzene. A comparison with the traditional AOCS titration method for oil analysis was performed. FFA content in mackerel frozen at -10 degrees C was measured according to the three methods over a 12 month period. The results showed similar values, and good correlations were obtained.