Effects of Package Atmosphere and Storage Conditions on Minimizing Risk of Escherichia coli O157:H7 in Packaged Fresh Baby Spinach.

Packaged fresh spinach has been associated with outbreaks of illness caused by Escherichia coli O157:H7. The purpose of this study was to assess the behavior of E. coli O157:H7 in packaged baby spinach in response to storage conditions of temperature and package atmosphere and including effects of inoculation level, spinach leaf damage (cut leaves), internalized or leaf surface contamination, exposure to hypochlorite sanitizer, and package size. Behavior of E. coli O157:H7 inoculated at 2 and 4 log CFU/g on spinach packaged in polymer bags composed of a two-layer laminate (polypropylene and polyethylene) and stored under atmospheres of 20% O2-3% CO2 and 0% O2-15% CO2 (aerobic and anaerobic, respectively) was assessed at 5, 7, 12, and 15°C for up to 14 days. Growth kinetics were calculated using DMFit software. Temperature decreases progressively diminished growth or survival of the pathogen, and an aerobic package atmosphere resulted in longer lag times (4 to 6 days) and lower population levels (0.2 to 1.4 log CFU/g) compared with the anaerobic atmosphere at 15°C. Internalized contamination, leaf cuts, or exposure to 100 ppm of hypochlorite did not result in changes in pathogen behavior compared with controls; however, a growth minimization trend consisting of longer lag times and lower population levels was repeatedly observed in the aerobic compared with the anaerobic package atmospheres. In contrast, growth of indigenous mesophiles and Enterobacteriaceae was unaffected by package atmosphere. Spinach stored at 5 to 7°C in two sizes (5 and 16 oz) of polyethylene terephthalate clamshell packages with ambient air atmospheres was more likely to progress to lower-oxygen conditions in 16-oz compared with 5-oz packages after 7 days of storage (P < 0.05). Practices to maintain aerobic conditions within the package, as well as storage of the package at low temperature, are ways to limit growth of E. coli O157:H7 in packaged spinach.

Optimization of lactulose synthesis from whey lactose by immobilized ß-galactosidase and glucose isomerase.

In the present study, commercially available whey was used as a lactose source, and immobilized ß-galactosidase and glucose isomerase were used to synthesize lactulose from whey lactose in the absence of fructose. Optimal reaction conditions, such as lactose concentration, temperature, ionic strength of the buffer, and ratio of immobilized enzymes, were determined to improve lactulose synthesis using immobilized enzymes. Lactulose synthesis using immobilized enzymes improved markedly after optimizing the reaction conditions. When the lactulose synthesis was carried out at 53.5°C using 20% (w/v) whey lactose, 12U/ml of immobilized ß-galactosidase and 60U/ml of immobilized glucose isomerase in 100mM sodium phosphate buffer at pH 7.5, the lactulose concentration and specific productivity were 7.68g/l and 0.32mg/Uh, respectively. Additionally, when the immobilized enzymes were reused for lactulose synthesis, their catalytic activity was 57.1% after 7 repeated uses.

Batch and continuous synthesis of lactulose from whey lactose by immobilized ß-galactosidase.

In this study, lactulose synthesis from whey lactose was investigated in batch and continuous systems using immobilized ß-galactosidase. In the batch system, the optimal concentration of fructose for lactulose synthesis was 20%, and the effect of galactose, glucose and fructose on ß-galactosidase activity was determined for hydrolysis of whey lactose and the transgalactosylation reaction for lactulose synthesis. Galactose and fructose were competitive inhibitors, and glucose acted as a noncompetitive inhibitor. The inhibitory effects of galactose and glucose were stronger in the transgalactosylation reaction than they were in the hydrolysis reaction. In addition, when immobilized ß-galactosidase was reused for lactulose synthesis, its catalytic activity was retained to the extent of 52.9% after 10 reuses. Lactulose was synthesized continuously in a packed-bed reactor. We synthesized 19.1g/l lactulose during the continuous flow reaction at a flow rate of 0.5 ml/min.

Biodiesel production by a mixture of Candida rugosa and Rhizopus oryzae lipases using a supercritical carbon dioxide process.

In this study, various factors, such as temperature, pressure, agitation speed, water content, and the concentration and ratio of immobilized ROL and CRL were investigated for the efficient enzymatic production of biodiesel using a supercritical carbon dioxide process. Furthermore, a stepwise reaction method for the maintenance of immobilized lipase activity was optimized. Optimal conditions for biodiesel production were determined to be as follows: 130 bar pressure, 45 °C temperature, 250 rpm agitation speed, 10% water content, and 20% immobilized ROL and CRL (1:1). When batch process was performed under optimal conditions, the biodiesel conversion yield was 99.13% at 3 h. Biodiesel conversion yield was 99.99% at 2 h when 90 mmol methanol was used in a stepwise reaction. Moreover, the conversion yield of biodiesel produced by the repeated recycling of immobilized lipase in the stepwise reactions was 85% after 20 reuses.

Quantitative detection of glyphosate by simultaneous analysis of UV spectroscopy and fluorescence using DNA-labeled gold nanoparticles.

A sandwich-type immunosensor composed of antigen-double target/probe DNA-coated gold nanoparticles (NPs) was developed for the measurement of fluorescence intensity and quantitative analysis of single-stranded DNA based on the concentration of free glyphosate. The reaction between the antigen-double DNA-gold NPs and immobilized antibody on the substrate was carried out for 2 h. The results of the antigen-antibody reaction were measured on the basis of the fluorescence intensity obtained from comparison with the free antigens at concentrations of 0.01-100 µg mL(-1) for the detection of immobilized antigen-double DNA-gold NPs. For the quantitative analysis based on the concentration of glyphosate(0.01-100 µg mL(-1)), the immunosensor response also revealed the same detection range of glyphosate using DNA detection.

The within-host population dynamics of normal flora in the presence of an invading pathogen and antibiotic treatments.

A mathematical competition model between normal flora and an invading pathogen was devised to allow analysis of bacterial infections in a host. The normal flora includes the various microorganisms that live on or within the host and act as a primary human immune system. Despite the important role of the normal flora, no mathematical study has been undertaken on models of the interaction between it and invading pathogens against a background of antibiotic treatment. To quantify key elements of bacterial behavior in a host, pairs of nonlinear differential equations were used to describe three categories of human health conditions, namely, healthy, latent infection, and active infection. In addition, a cutoff value was proposed to represent the minimum population level required for survival. The recovery of normal flora after antibiotic treatment was also included in the simulation because of its relation to human health recovery. The significance of each simulation parameter for the bacterial growth model was investigated. The devised simulation showed that bacterial proliferation rate, carrying capacity, initial population levels, and competition intensity have a significant effect on bacterial behavior. Consequently, a model was established to describe competition between normal flora and an infiltrating pathogen. Unlike other population models, the recovery process described by the devised model can describe the human health recovery mechanism.

Fatty acids reduce the tensile strength of fungal hyphae during cephalosporin C production in Acremonium chrysogenum.

Fragmentation rate constants, which can be used to estimate the tensile strength of fungal hyphae, were used to elucidate relationships between morphological changes and addition of fatty acids during cephalosporin C production in Acremonium chrysogenum M35. The number of arthrospores increased gradually during fermentation, and, in particular, was higher in the presence of rice oil, oleic acid or linoleic acid than in their absence. Because supplementation of rice oil or fatty acids increased cephalosporin C, we concluded that differentiation to arthrospores is related to cephalosporin C production. To estimate the relative tensile strengths of fungal hyphae, fragmentation rate constants (k (frag)) were measured. When rice oil, oleic acid, or linoleic acid were added into medium, fragmentation rate constants were higher than for the control, and hyphal tensile strengths reduced. The relative tensile strength of fungal hyphae, however was not constant presumably due to differences in physiological state.

Optimization of culture medium for lactosucrose ( G-beta-D-galactosylsucrose) Production by Sterigmatomyces elviae mutant using statistical analysis.

In this study, the optimization of culture medium using a Sterigmatomyces elviae mutant was investigated using statistical analysis to increase the cell mass and lactosucrose ((4)G-beta-D-galactosylsucrose) production. In basal medium, the cell mass and lactosucrose production were 4.12 g/l and 140.91 g/l, respectively. However, because of the low cell mass and lactosucrose production, optimization of culture medium was carried out to increase the cell mass and lactosucrose production. Culture media were optimized by the S. elviae mutant using analysis of variance (ANOVA) and response surface methodology (RSM). Central composite designs using RSM were utilized in this investigation. Quadratic models were obtained for cell mass and lactosucrose production. In the case of cell mass, optimal components of the medium were as follows: sucrose 1.13%, yeast extract 0.99%, bactopeptone 2.96%, and ammonium sulfate 0.40%. The predicted maximum value of cell mass was about 5.20 g/l and its experimental value was 5.08 g/l. In the case of lactosucrose production, optimal components of the medium were as follows: sucrose 0.96%, yeast extract 1.2%, bactopeptone 3.0%, and ammonium sulfate 0.48%. Then, the predicted maximum value of lactosucrose production was about 194.12 g/l and the corresponding experimental value was about 183.78 g/l. Therefore, by culturing using predicted conditions, the real cell mass and lactosucrose production increased to 23.3% and 30.42%, respectively.