The taxonomy of Enterobacter sakazakii: proposal of a new genus Cronobacter gen. nov. and descriptions of Cronobacter sakazakii comb. nov. Cronobacter sakazakii subsp. sakazakii, comb. nov., Cronobacter sakazakii subsp. malonaticus subsp. nov., Cronobacter turicensis sp. nov., Cronobacter muytjensii sp. nov., Cronobacter dublinensis sp. nov. and Cronobacter genomospecies 1

BACKGROUND: Enterobacter sakazakii is an opportunistic pathogen that can cause infections such as necrotizing enterocolitis, bacteraemia, meningitis and brain abscess/lesions. When the species was defined in 1980, 15 biogroups were described and it was suggested that these could represent multiple species. In this study the taxonomic relationship of strains described as E. sakazakii was further investigated. RESULTS: Strains identified as E. sakazakii were divided into separate groups on the basis of f-AFLP fingerprints, ribopatterns and full-length 16S rRNA gene sequences. DNA-DNA hybridizations revealed five genomospecies. The phenotypic profiles of the genomospecies were determined and biochemical markers identified. CONCLUSION: This study clarifies the taxonomy of E. sakazakii and proposes a reclassification of these organisms

Development of a novel smoke-flavoured salmon product by sodium replacement using water vapour permeable bags

This is the peer reviewed version of the following article: Rizo Parraga, Arancha Maria, Fuentes López, Ana, Barat Baviera, José Manuel, Fernández Segovia, Isabel. (2018). Development of a novel smoke-flavoured salmon product by sodium replacement using water vapour permeable bags.Journal of the Science of Food and Agriculture, 98, 7, 2721-2728. DOI: 10.1002/jsfa.8767, which has been published in final form at http://doi.org/ 10.1002/jsfa.8767. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.[EN] BACKGROUND: Food manufacturers need to reduce sodium contents to meet consumer and public health demands. In this study the use of sodium-free (SF) salt and KCl to develop a novel smoke-flavoured salmon product with reduced sodium content was evaluated. Fifty percent of NaCl was replaced with 50% of SF salt or 50% KCl in the salmon smoke-flavouring process carried out using water vapour permeable bags. RESULTS: Triangle tests showed that samples with either SF salt or KCl were statistically similar to the control samples (100% NaCl). Since no sensorial advantage in using SF salt was found compared with KCl and given the lower price of KCl, the KCl-NaCl samples were selected for the next phase. The changes of physicochemical and microbial parameters in smoke-flavoured salmon during 42 days showed that partial replacement of NaCl with KCl did not significantly affect the quality and shelf-life of smoke-flavoured salmon, which was over 42 days. CONCLUSION: Smoke-flavoured salmon with 37% sodium reduction was developed without affecting sensory features and shelf-life. This is an interesting option for reducing sodium content in such products to help meet the needs set by both health authorities and consumers.We gratefully acknowledge the support of Tub-Ex Aps (Taars, Denmark) for suppling the water vapour permeable bags and for providing all of the necessary technical information. Arantxa Rizo thanks the Universitat Politecnica de Valencia for the FPI grant.Rizo Parraga, AM.; Fuentes López, A.; Barat Baviera, JM.; Fernández Segovia, I. (2018). Development of a novel smoke-flavoured salmon product by sodium replacement using water vapour permeable bags. Journal of the Science of Food and Agriculture. 98(7):2721-2728. https://doi.org/10.1002/jsfa.8767S2721272898

Comparing the antimicrobial effectiveness of NaCl and KCl with a view to salt/sodium replacement.

A study using a small range of pathogenic bacterial species (Aeromonas hydrophila, Enterobacter sakazakii, Shigella flexneri, Yersinia enterocolitica and 3 strains of Staphylococcus aureus) has shown that potassium chloride has an equivalent antimicrobial effect on these organisms when calculated on a molar basis. Combined NaCl and KCl experiments were carried out and data was analysed using a modification to the Lambert and Lambert [Lambert, R.J.W., and Lambert, R., 2003. A model for the efficacy of combined inhibitors. Journal of Applied Microbiology 95, 734–743.] model for combined inhibitors and showed that in combination KCl is a direct 1:1 molar replacement for the antimicrobial effect of common salt. If this is a general finding then, where salt is used to help preserve a product, partial or complete replacement by KCl is possible

An explanation for the effect of inoculum size on MIC and the growth/no growth interface.

The inoculum effect (IE) is the phenomenon observed where changes in the inoculum size used in an experiment alters the outcome with respect to, for example, the minimum inhibitory concentration of an antimicrobial or the growth/no growth boundary for a given set of environmental conditions. Various hypotheses exist as to the cause of the IE such as population heterogeneity and quorum sensing, as well as the null hypothesis — that it is artefactual. Time to detection experiments (TTD) were carried out on different initial inoculum sizes of several bacterial species (Aeromonas hydrophila, Enterobacter sakazakii, Salmonella Poona, Escherichia coli and Listeria innocua) when challenged with different pH and with combined pH and sodium acetate. Data were modelled using a modification to a Gamma model (Lambert and Bidlas 2007, Int. J. Food Microbiology 115, 204–213), taking into account the inoculum size dependency on the TTD obtained under ideal conditions. The model suggests that changes in minimum inhibitory concentration (MIC) or in the growth–no growth boundary with respect to inoculum size are due to using a smaller or larger inoculum (i.e. is directly related to microbial number) and is not due to other, suggested, phenomena. The model used further suggests that the effect of a changing inoculum size can be modelled independently of any other factor, which implies that a simple 1 to 2-day experiment measuring the TTD of various initial inocula can be used as an adjunct to currently available models

Gamma Study of pH, Nitrite, and Salt Inhibition of Aeromonas hydrophila

The gamma hypothesis states that there are no interactions between antimicrobial environmental factors. The time to growth of Aeromonas hydrophila challenged with pH, NaNO(2), and salt combinations at 30°C was investigated. Data were examined using a model based on the gamma hypothesis (the gamma model), which takes into account variance-stabilizing transformations and which gives biologically relevant parameters. At high concentrations of NaNO(2) and at pHs of >6.0, the antimicrobial action of the nitrite ion has a strong influence (MIC = 2,033 mg liter(−1)), whereas at pHs of <6, nitrous acid is dominant (MIC = 1.5 mg liter(−1)). This change is not due to a “synergy” between pH and the nitrite ion but is due to the shift in the equilibrium concentrations of nitrous acid and nitrite in solution caused by pH. In combination with salt, the parameters found for the action of Na nitrite were identical to those found when it was examined in isolation. Therefore, pH, NaNO(2), and salt act independently on the growth of A. hydrophila. By expanding the gamma model with a cardinal temperature model, the results of fitting the model of Palumbo et al. (J. Food Prot. 54:429-435, 1994) to randomly produced environmental conditions could be reproduced, suggesting that temperature also has an independent effect