Núria Rius

Flow cytometry applications in the food industry.

Flow cytometry has become a valuable tool in food microbiology. By analysing large numbers of cells individually using light-scattering and fluorescence measurements, this technique reveals both cellular characteristics and the levels of cellular components. Flow cytometry has been developed to rapidly enumerate microorganisms; to distinguish between viable, metabolically active and dead cells, which is of great importance in food development and food spoilage; and to detect specific pathogenic microorganisms by conjugating antibodies with fluorochromes, which is of great use in the food industry. In addition, high-speed multiparametric data acquisition, analysis and cell sorting, which allow other characteristics of individual cells to be studied, have increased the interest of food microbiologists in this technique. This mini-review gives an overview of the principles of flow cytometry and examples of the application of this technique in the food industry.

The role of pH in the ‘glucose effect’ on prodigiosin production by non‐proliferating cells of Serratia marcescens

The production of prodigiosin by non‐proliferating cells of Serratia marcescens is inhibited by addition of glucose or different carbon sources to the induction medium. The induction in acidic external pH, mimicking the effects produced by the carbon sources, reduced prodigiosin synthesis, and the prodigiosin production seems to be related to the length of the low pH period. Buffering at pH 7·5 increased pigment production in media with repressing carbon sources. This study reveals that the inhibitory effect of carbon sources on prodigiosin production may be due to a lowering of the pH of the medium.

The effect of pH on prodigiosin production by non‐proliferating cells of Serratia marcescens

The synthesis of prodigiosin by non‐proliferating cells of Serratia marcescens was examined at various pH values between 5.5 and 9.5. During incubation in unbuffered medium, pH changed and prodigiosin production was similar regardless of the initial pH. Variations in pigment production were noted when buffers were employed in cultures of non‐proliferating cells. The optimum pH for prodigiosin production was 8.0–8.5. Proline oxidase was also measured. The results suggest that the effect of pH may be related to the amount of proline which can be incorporated into prodigiosin.

Inhibitory activity of monofloral and multifloral honeys against bacterial pathogens

Summary Thirty one honey samples were evaluated for their ability to inhibit the growth of ten bacterial pathogens: Escherichia coli O157:H7; Salmonella enterica ser. Typhimurium; Shigella sonnei; Pseudomonas aeruginosa; Bacillus cereus; Listeria monocytogenes; Staphylococcus aureus 9144 and MRSA; Streptococcus mutans; and Enterococcus faecalis, using a well diffusion assay. All Gram-negative bacteria except P. aeruginosa were sensitive to all of the honey samples and also to artificial honey used as control at 25% concentration. S.sonnei was the pathogen whose growth was most inhibited by honey. P. aeruginosa exhibited overgrowth of surrounding wells inoculated with five honey samples. Each Gram-positive pathogen and the different strains of S. aureus exhibited different sensitivities to honeys. The three honeys from Galicia (Spain) were the most active. E. coli O157:H7, S. sonnei, MRSA, S. mutans and E. faecalis were studied for specific inhibitory activity. E. coli O157:H7, S. sonnei and E faecalis were inhibited by high sugar concentration. The inhibition of MRSA and S. mutans was caused by hydrogen peroxide generation. MRSA was also inhibited by two unprocessed honeys due to the presence of proteinaceous compounds. Such activity is potentially of importance in honey used as wound dressing and also as a preservative in minimally processed foods.

Effect of Tocopherol Extract, Staphylococcus carnosus Culture, and Celery Concentrate Addition on Quality Parameters of Organic and Conventional Dry-Cured Sausages

The effects of the addition to sausage mix of tocopherols (200 mg/kg), a conventional starter culture with or without Staphylococcus carnosus, celery concentrate (CP) (0.23% and 0.46%), and two doses of nitrate (70 and 140 mg/kg expressed as NaNO(3)) on residual nitrate and nitrite amounts, instrumental CIE Lab color, tocol content, oxidative stability, and overall acceptability were studied in fermented dry-cured sausages after ripening and after storage. Nitrate doses were provided by nitrate-rich CP or a chemical grade source. The lower dose complies with the EU requirements governing the maximum for ingoing amounts in organic meat products. Tocopherol addition protected against oxidation, whereas the nitrate dose, nitrate source, or starter culture had little influence on secondary oxidation values. The residual nitrate and nitrite amounts found in the sausages with the lower nitrate dose were within EU-permitted limits for organic meat products and residual nitrate can be further reduced by the presence of the S. carnosus culture. Color measurements were not affected by the CP dose. Product consumer acceptability was not affected negatively by any of the factors studied. As the two nitrate sources behaved similarly for the parameters studied, CP is a useful alternative to chemical ingredients for organic dry-cured sausage production.

Effect of Fermentation Time and Vegetable Concentrate Addition on Quality Parameters of Organic Botifarra Catalana, a Cured–Cooked Sausage

The effects of the addition of two different sources of nitrite (pure NaNO2 or a nitrate-rich vegetable concentrate) and three different fermentation times with nitrate-reducing cultures (6, 12, or 24 h at 16 °C) on microbial counts, pH, residual nitrate and nitrite amounts, and susceptibility to oxidation of botifarra catalana sausage were studied. Moreover, curing efficiency, color, tocopherol and tocotrienol contents, oxidative status, and consumer acceptability of these sausages were assessed after vacuum packaging and storage at 4 °C for up to 180 days. Residual nitrate and nitrite amounts were lower than the limits established by the European Union for organic meat products. Longer periods of fermentation produced higher meat curing efficiency ratios, whereas consumer acceptability scores were highest for sausages with added vegetable concentrate. Storage of the sausages caused small quality changes. Therefore, these results indicate that vegetable concentrate is a useful alternative for organic cured-cooked meat products.

Buffering capacity and H+ membrane conductance of Gram-negative bacteria

Buffering capacity and membrane H+ conductance were examined in seven Gram-negative species: Aquaspirillum serpens, Pseudomonas aeruginosa, Alcaligenes faecalis, Escherichia coli, Salmonella typhimurium, Proteus mirabilis and Aeromonas hydrophila. All strains of Enterobacteriaceae studied here showed a decrease in both parameters as the external pH increased, over the pH range studied. The other four species presented an increase in buffering capacity and membrane conductance to protons as the external pH increased from 5.5 to 7.0.

Acid-base response of bacterial suspensions

The response of suspensions of non-proliferating bacteria to external pH ranging from 3.6 to 9.9 was examined. The acid-base response is affected by the number of viable cells in the suspensions and culture conditions (aerobiosis or anaerobiosis) but not by culture time, composition of the culture medium or the age of suspensions. The results with carbonylcyanide m-chlorophenylhydrazone-and N, N′- dicyclohexylcarbodiimide-treated cells indicated that passive proton conductance and proton translocation by ATP could be involved in this response. pH profiles of suspensions of 18 chemoheterotrophic Gram-negative and Gram-positive bacteria support the hypothesis that the proton gradient generated by an alkali or an acid shift could be dissipated by bacteria with respiratory metabolism.

Buffering capacity and membrane H+ conductance ofZymomonas mobilis

Buffering power and membrane conductance to H+ were measured inZymomonas mobilis subspmobilis ATCC 29191 by a pulse technique. Over the pH range studied, from 4.02 to 7.44,Z. mobilis presented very high values of cytoplasmic buffering capacity; it was a significant proportion of the total buffering capacity. These results support the idea that the cytoplasmic buffering power might be part of the pH homeostatic mechanism.

Buffering capacity and membrane H+ conductance of acetic acid bacteria

SummaryBuffering capacities and membrane conductance to H+ were measure inAcetobacter aceti ATCC 15973 andGluconobacter oxydans ATCC 621 by a pulse technique. In both strains the buffering capacity of intact cells was a significant proportion of the total buffering capacity, but the magnitude of the buffering capacity varied between one species and another. Over the pH range studied, 4.02 to 8.15,Gluconobacter oxydans, which oxidizes sugars and alcohols to acids and accumulates them, showed lower values of buffering capacities and membrane conductance to protons thanAcetobacter aceti, which oxidizes these substrates completely to CO2 and H2O.