Outgrowth inhibition of Clostridium beijerinckii spores by a bacteriocin-producing lactic culture in ovine milk cheese

In the manufacture of model cheeses, ovine milk was deliberately contaminated with spores of Clostridium beijerinckii INIA 63, a wild isolate from Manchego cheese with late blowing defect, and inoculated with nisin- and lacticin 481-producing Lactococcus lactis subsp. lactis INIA 415 as starter, to test its potential to prevent the late blowing defect, or with L. lactis subsp. lactis INIA 415-2, a spontaneous mutant not producing bacteriocins. Cheeses made individually with the lactococcal strains, without clostridial spores, served as controls. Cheese made with clostridial spores and L. lactis subsp. lactis INIA 415-2 showed late blowing defect after 120 days of ripening. Spoilt cheese also showed lower concentrations of lactic acid, and higher levels of acetic, propionic and butyric acids, and of other volatile compounds such as 2-propanol and 1-butanol, than control cheese. In addition, cheese made with the bacteriocin producer did not show any late blowing symptoms, despite its spore counts similar to those of blown cheese, pointing to outgrowth inhibition of C. beijerinckii spores by bacteriocins. Besides, cheese made with the bacteriocin producer showed similar concentrations of lactic acid and volatile compounds than control cheese. Inclusion of L. lactis subsp. lactis INIA 415 in starter cultures seems a feasible method to prevent late blowing defect in cheese without altering its sensory characteristics.     

Optimization, Modeling, and Online Monitoring of the Enzymatic Extraction of Banana Juice

This article focuses on the optimization, modeling, and online monitoring of banana juice production through an enzymatic method. In order to perform this task, a batch reactor was designed with automatic control over the temperature and the agitation speed as well as online monitoring of torque. The experiments were carried out with the Musa AAA Cavendish banana variety (Enano gigante), the main variety planted in Mexico. Three different ripening stages were evaluated. Optimization of juice extraction was evaluated as a function of the pulp/water relationship and the concentration of the enzyme complex. The results showed that the adding of water had no influence on the extraction of banana juice, and the optimal enzyme concentration per kilogram of banana pulp was found. Based on a fuzzy logic approach, it was possible to relate the initial torque with the ripeness stage. Furthermore, an observable dynamical model based on ordinary differential equations and fuzzy logic is presented. With this model, the relationship between the torque dynamic and the instant juice yield was found to depend on the amount of enzyme, the temperature, and the maturity stage of the banana used. In addition, a principal components analysis was used to classify and to relate the final juice characteristics (e.g., L, a, and b colorimetric components) to the processing conditions and the final appreciation of a group of sensorial panelists. Additionally, a robust observer was designed and implemented to filter the noise present in the torque signal and to predict the instant juice yield.     

Using High-Pressure Processing for Reduction of Proteolysis and Prevention of Over-ripening of Raw Milk Cheese

High-pressure-processing (HPP) at 400 or 600 MPa was applied to cheeses made from ewe raw milk, on days 21 or 35 after manufacturing, to reduce proteolysis and prevent over-ripening. The characteristics of HPP and non-pressurized (control) cheeses were compared during ripening at 8 °C until day 60 and further storage at 4 °C until day 240. HPP and control cheeses showed similar pH values throughout ripening, but on day 240 pH values remained 0.4–0.6 units lower for HPP cheeses than for the control cheeses. Casein degradation was significantly retarded in the 600 MPa cheeses. Their α-casein concentration was 48–52 % higher on day 60 and 30–33 % higher on day 240 than in the control cheeses while β-casein concentration was 25–26 % higher on day 60 and 100–103 % higher on day 240. No significant differences in para-κ-casein concentration between cheeses were found on day 60, but on day 240, it was 22–35 % higher in the 600 MPa cheeses than in the control cheese. Hydrophilic peptides, hydrophobic peptides and total free amino acids evolved similarly in HPP and control cheeses during the 60-day ripening period. However, on day 240 hydrophilic peptides were at 34–39 % lower levels in the 600 MPa cheeses than in the control cheeses, hydrophobic peptides at 7–16 % lower levels and total free amino acids at 25–29 % lower levels. Flavour intensity scores increased at a slower rate in HPP cheeses than in the control cheese. Flavour quality declined markedly in the control cheeses during refrigerated storage while it did not vary significantly in 600 MPa cheeses.     

Reuterin and High Hydrostatic Pressure Treatments on the Inactivation of Listeria monocytogenes and Effect on the Characteristics of Cold-Smoked Salmon

The effect of reuterin and high hydrostatic pressure (HHP) processing at 450 MPa for 5 min on the inactivation of Listeria monocytogenes and the characteristics of cold-smoked salmon during 35 days at 4 and 10 °C were investigated. The growth rate of the pathogen was reduced by reuterin addition and a synergistic antimicrobial effect against L. monocytogenes was recorded when the biopreservative was applied in combination with HHP at 450 MPa for 5 min. This combined treatment prevented the pathogen recovery observed with individual treatments and delayed the spoilage of smoked salmon maintaining total viable counts under 3.5 log units during 35 days of storage at 4 °C. All treatments assayed induced changes in lightness (L*) and redness (a*), resulting in a brighter appearance of smoked salmon, whereas no modifications were recorded in shear strength values immediately after treatments. Moreover, reuterin and HHP treatments, individually or in combination, avoided the formation of biogenic amines during the 35 days of storage at 4 and 10 °C. The addition of reuterin in combination with HHP at 450 MPa for 5 min might be applied as a hurdle technology to improve the safety and extend the shelf life of lightly preserved seafood products, such as cold-smoked salmon.     

Water activity,water glass dynamics,and the control of microbiological growth in foods

Water is probably the single most important factor governing microbial spoilage in foods, and the concept of water activity (a_w) has been very valuable because measured values generally correlate well with the potential for growth and metabolic activity. Despite some drawbacks (e.g., solute effect), the concept of a_w has assisted food scientists in their effort to predict the onset of food spoilage as well as to control food‐borne disease hazards in food products. In the last decade the concept of a_w has been challenged. It has been suggested that reduced‐moisture food products (e.g., low and intermediate) may be nonequilibrium systems and that most of them are in the amorphous metastable state, which is very sensitive to changes in moisture content and temperature. It has been proposed that the glass transition temperature T_g (temperature at which the glass‐rubber transition occurs), is a parameter that can determine many product properties, the safety of foods among them. The concept of water dynamics, originating in a food polymer science approach, has been suggested instead of a_w to better predict the microbial stability of intermediate‐moisture foods. The usage of a_w to predict microbial safety of foods has been discouraged on the basis that (1) in intermediate‐moisture foods the measured water vapor pressure is not an equilibrium one, and because a_w is a thermodynamic concept, it refers only to equilibrium; and (2) the microbial response may differ at a particular a_w when the latter is obtained with different solutes.

This review analyzes these suggestions on the basis of abundant experimental evidence found in the literature. It is concluded that nonequilibrium effects (e.g., inability of water to diffuse in a semimoist food) appear to be in many cases slow within the time frame (food's shelf life) of the experiments and/or so small that they do not affect seriously the application of the a_w concept as a predictor of microbial stability in foods.

The claims that a food science polymer approach to understanding the behavior of aqueous sugar glasses and concentrated solutions may be used to predict the microbial stability of food systems is not substantiated by experimental evidence. This approach does not offer, at the present time, a better alternative to the concept of a_w as a predictor of microbial growth in foods.

It is also recognized that a_w has several limitations and should be always used carefully, and this must include precautions regarding the possible influences of nonequilibrium situations. This aspect may be summarized by simply saying that anyone who is going to employ the term water activity must be aware of the implications of its definition.     

Competition mechanisms of lactic acid bacteria and bifidobacteria: Fermentative metabolism and colonization

Enzyme activities (α- and β-glucosidases, α- and β-galactosidases and β-fructofuranosidase) and organic acid production of four strains of lactic acid bacteria (LAB; Streptococcus thermophilus STY-31, Lactobacillus delbrueckii subsp. bulgaricus LBY-27, Lactobacillus casei LC-01 and Lactobacillus acidophilus LA-5) and Bifidobacterium lactis BB-12 were tested on milk and MRS fermentation broth with glucose, lactose or fructooligosaccharides (FOS) as carbon source. The highest β-galactosidase activity was found in L. acidophilus growing on milk. As compared to milk, α-glucosidase activity was increased with FOS by B. lactis, L. acidophilus and L. casei. The analysis of organic acids and short-chain fatty acids in the medium growth showed that lactate and acetate were the major fermentation metabolites produced by LAB and bifidobacteria, respectively. However, a metabolic shift towards more acetate and formate production, at the expense of lactate production, was observed during growth of L. casei on FOS. When grown on FOS as sole carbon source, L. acidophilus showed the highest production of lactate among the species tested. In addition, L. acidophilus demonstrated resistance to colonization against the intestinal pathogens Escherichia coli and Salmonella enterica in competition assays.     

Cultivation approach for comparing the nutritional quality of two pepper cultivars grown under different agricultural regimes

The aim of this study was to assess the influence of the cropping system (organic or conventional) and the growth medium (soil or soilless) on the antioxidant content of peppers from Almuden and Quito cultivars. This cultivation study complements a previous farm study. For both cultivars, organic cultivation led to higher fruit lipophilic antioxidant content than conventional management but differences in the hydrophilic fraction were not observed. The effect of the growth medium on pepper composition depended on the studied cultivar. In Almuden, neither the hydrophilic nor the lipophilic fractions were affected by the growth medium, while in Quito most compounds showed higher values in soil-grown plants. Taking into account the results from the present and the previous farm study we conclude that the effect of organic vs. conventional cultivation on pepper quality can be attributed to the different nutritional regimes and other factors associated with the agricultural system. However, both aspects are not necessarily different in organic and conventional commercial crops. The effect of the growth medium (soil or soilless) on pepper quality depended on the cultivar: Almuden was not affected by the growth medium and Quito was seen to be less suitable for soilless cultivation.     

Changes in the contents of antioxidant compounds in pepper fruits at different ripening stages,as affected by salinity

The interest in the consumption of pepper fruits (Capsicum annuum L.) is, to a large extent, due to its content of bioactive nutrients and their importance as dietary antioxidants. A greenhouse experiment was carried out to determine the effects of salinity and different ripening states of pepper fruits on several compounds with antioxidant properties. Fruits from plants grown under three saline treatments (0, 15, and 30 mM NaCl) were collected at three maturity states (green, turning, and red). Antioxidant activity in the hydrophilic (HAA) and lipophilic (LAA) fractions, lycopene, β-carotene, ascorbic acid, total phenolic compounds and reducing sugars were determined. From the nutritional point of view, the red state was the most appropriate state of maturation, since red peppers had the highest levels of lycopene, β-carotene, and sugars and the highest antioxidant activity for both hydrophilic and lipophilic fractions. The effect of salinity depended on the maturity state of the peppers: it had no effect on HAA, β-carotene or sugars, but decreased ascorbic acid and total phenolic compounds, and increased LAA and lycopene. The use of a moderately-saline water was beneficial when peppers were harvested in the red state, by increasing HAA and LAA in fruits, with no significant effects on other parameters.     

Polymer-coated nanoparticles: a universal tool for biolabelling experiments

Water solubilization of nanoparticles is a fundamental prerequisite for many biological applications. To date, no single method has emerged as ideal, and several different approaches have been successfully utilized. These 'phase-transfer' strategies are reviewed, indicating key advantages and disadvantages, and a discussion of conjugation strategies is presented. Coating of hydrophobic nanoparticles with amphiphilic polymers provides a generic pathway for the phase transfer of semiconductor, magnetic, metallic, and upconverting nanoparticles from nonpolar to polar environments. Amphiphilic polymers that include maleimide groups can be readily functionalized with chemical groups for specific applications. In the second, experimental part, some of the new chemical features of such polymer-capped nanoparticles are demonstrated. In particular, nanoparticles to which a pH sensitive fluorophore has been attached are described, and their use for intracellular pH-sensing demonstrated. It is shown that the properties of analyte-sensitive fluorophores can be tuned by using interactions with the underlying nanoparticles.     

Nutritional and health-related compounds in sprouts and seeds of soybean (<Emphasis Type="Italic">Glycine max</Emphasis>), wheat (<Emphasis Type="Italic">Triticum aestivum.L</Emphasis>) and alfalfa (<Emphasis Type="Italic">Medicago sativa</Emphasis>) treated by a new drying method

Nutritional and health-related compounds of alfalfa, wheat and soybean seeds dried by new drying process called the DIC process (controlled instantaneous pressure release) were evaluated, before and after sprouting. Vitamins (A, B₁, B₂, B₆, C and E), minerals (Fe, Mg, Ca, K, Mn, Na, Cu and Zn) and phytoestrogens (genistein and daidzein ) content were determined. Alfalfa, soybean and wheat seeds dried by DIC showed similar content of biological compounds to seeds dried by traditional processes. Sprouting DIC-seeds significantly increased the levels of vitamins, minerals and phytoestrogens, improving their nutrititional value and health quality compared to fresh products. The increase of 1250-fold and 10-fold of the initial vitamin A and vitamin C content, respectively, of alfalfa seeds due to sprouting is remarkable. Sprouts from DIC-seeds showed a significantly higher vitamin A content than sprouts obtained from seeds dried by other methods. Soybean sprouts obtained from DIC-seeds showed a significant increase in free phytoestrogens, quantified as genistein (70.34LJ.8 mg kg^-1 d.w.) and daidzein (109.17.3ᆥ.3 mg kg^-1 d.w.).