Impact of enriched environment on motor performance and learning in mice.

Neuroscience heavily relies on animal welfare in laboratory rodents as it can significantly affect brain development, cognitive function and memory formation. Unfortunately, laboratory animals are often raised in artificial environments devoid of physical and social stimuli, potentially leading to biased outcomes in behavioural assays. To assess this effect, we examined the impact of social and physical cage enrichment on various forms of motor coordination. Our findings indicate that while enriched-housed animals did not exhibit faster learning in eyeblink conditioning, the peak timing of their conditioned responses was slightly, but significantly, improved. Additionally, enriched-housed animals outperformed animals that were housed in standard conditions in the accelerating rotarod and ErasmusLadder test. In contrast, we found no significant effect of enrichment on the balance beam and grip strength test. Overall, our data suggest that an enriched environment can improve motor performance and motor learning under challenging and/or novel circumstances, possibly reflecting an altered state of anxiety.

Membrane permeabilization and cellular death of Escherichia coli, Listeria monocytogenes and Saccharomyces cerevisiae as induced by high pressure carbon dioxide treatment.

In this study, the relationship between (irreversible) membrane permeabilization and loss of viability in Escherichia coli, Listeria monocytogenes and Saccharomyces cerevisiae cells subjected to high pressure carbon dioxide (HPCD) treatment at different process conditions including temperature (35-45 degrees C), pressure (10.5-21.0 MPa) and treatment time (0-60 min) was examined. Loss of membrane integrity was measured as increased uptake of the fluorescent dye propidium iodide (PI) with spectrofluorometry, while cell inactivation was determined by viable cell count. Uptake of PI by all three strains indicated that membrane damage is involved in the mechanism of HPCD inactivation of vegetative cells. The extent of membrane permeabilization and cellular death increased with the severity of the HPCD treatment. The resistance of the three tested organisms to HPCD treatment changed as a function of treatment time, leading to significant tailing in the survival curves, and was dependent on pressure and temperature. The results in this study also indicated a HPCD-induced damage on nucleic acids during cell inactivation. Transmission electron microscopy showed that HPCD treatment had a profound effect on the intracellular organization of the micro-organisms and influenced the permeability of the bacterial cells by introducing pores in the cell wall.

The development of Escherichia coli and Listeria monocytogenes variants resistant to high-pressure carbon dioxide inactivation.

AIMS: The objective of this study was to investigate whether bacterial cells could develop resistance (as a part of their adaptation strategy) to high-pressure CO(2) (HPCD) inactivation. METHODS AND RESULTS: Alternating cycles of exposure to pressurized CO(2) (10.5 MPa, 35 degrees C, 400 min(-1), 70% working volume ratio during 10 min) and re-growth of the surviving subpopulation were used to investigate possible increases in the resistance of Escherichia coli and Listeria monocytogenes to HPCD. The results show an increased resistance of both pathogens tested after seven cycles of inactivation. Increase in the resistance after 15 cycles resulted in a difference of 2.4 log CFU ml(-1) in log N(0)/N(i) when parental (N(0)) and treated cultures (N(i)) of E. coli and L. monocytogenes were compared. CONCLUSIONS: Current findings indicate the ability of micro-organisms to adapt to HPCD preservation technology. SIGNIFICANCE AND IMPACT OF THE STUDY: The occurrence of HPCD-resistant micro-organisms could pose a new hazard to the safety and stability of HPCD-processed foods.

Influence of type of microorganism, food ingredients and food properties on high-pressure carbon dioxide inactivation of microorganisms.

High pressure carbon dioxide (HPCD) treatment is currently considered as an attractive non-thermal process for preserving food. Industrial application of this technique requires, among others, systematic (quantitative) data on the inactivation of food relevant pathogenic and spoilage microorganisms, and in-depth information on the effect that the composition and the properties of a food matrix have on the inactivation efficacy. The first objective of this study, therefore, is to evaluate and compare the HPCD susceptibility of several food pathogens and spoilage microorganisms under the same treatment conditions. In the second part, the influence of different food components (NaCl, oil, starch, whey protein and emulsifier) and food properties (pH, fluid viscosity and water activity) on the inactivation efficacy of HPCD was determined. For the first aim, a range of Gram-negative and Gram-positive bacteria, yeasts and spores were treated with pressurized CO(2) at 10.5 MPa and 35 degrees C during 20 min. Bacterial susceptibility towards HPCD treatments followed the sequence Gram-negative approximately Gram-positive>yeasts>spores and appeared to be related to the acid resistance of the organisms. To study the effect of different food compounds on HPCD inactivation, the reduction degree of Pseudomonas fluorescens was determined in media with and without these components at 10.5 MPa and 35 degrees C after 5 or 20 min, depending on the tested component. NaCl and the emulsifiers Tween 80 and sucrose stearate enhanced bacterial reduction, while oil reduced the bactericidal efficacy of HPCD. Starch and whey proteins did not influence inactivation. Finally, the influence of pH, fluid viscosity and water activity was investigated by determining the reduction of P. fluorescens at 10.5 MPa and 35 degrees C in suspensions from which the pH, viscosity and water activity were adjusted with respectively NaOH or HCl, gelatin or polyethylene glycol, and sucrose, NaCl or glycerol. Treatment time depended on the studied food property with 5 min for the pH experiments, while other experiments lasted 20 min. The results indicated that P. fluorescens cells became more sensitive to HPCD treatments at low pH and viscosity. Not water activity but the kind of soluble solute used to lower water activity influenced inactivation. High NaCl-concentrations lead to total inactivation, while sucrose and glycerol strongly protected the cells against inactivation.

High pressure carbon dioxide inactivation of microorganisms in foods: the past, the present and the future.

Thermal pasteurization is a well known and old technique for reducing the microbial count of foods. Traditional thermal processing, however, can destroy heat-sensitive nutrients and food product qualities such as flavor, color and texture. For more than 2 decades now, the use of high-pressure carbon dioxide (HPCD) has been proposed as an alternative cold pasteurization technique for foods. This method presents some fundamental advantages related to the mild conditions employed, particularly because it allows processing at much lower temperature than the ones used in thermal pasteurization. In spite of intensified research efforts the last couple of years, the HPCD preservation technique has not yet been implemented on a large scale by the food industry until now. This review presents a survey of published knowledge concerning the HPCD technique for microbial inactivation, and addresses issues of the technology such as the mechanism of carbon dioxide bactericidal action, the potential for inactivating vegetative cells and bacterial spores, and the regulatory hurdles which need to be overcome. In addition, the review also reflects on the opportunities and especially the current drawbacks of the HPCD technique for the food industry.

Bioavailability of Cd to the common carp, Cyprinus carpio, in the presence of humic acid.

The uptake of cadmium by the common carp, Cyprinus carpio, was studied in chemically defined freshwater in the absence and presence of commercial humic acid. This was done to evaluate whether the cadmium uptake by carp in the presence of humic acid was related to the ambient Cd2+ -ion activity or whether the complexed metal also contributed to the uptake. Uptake of Cd during a 3-h period of exposure was used as a measure of the biological availability of the metal. The uptake rate data for Cd in total fish and gills obtained in the absence (control) and presence (treatment) of humic acid were analyzed using a Michaelis-Menten model for mediated transport. The Michaelis-Menten parameters KM and Vmax obtained in the control and the treatment experiment were compared for each of the two investigated carp compartments (total fish and gills). The model parameter estimates for Cd uptake by total carp in the treatment experiment (KM = 0.41 +/- 0.11 micromol l(-1); Vmax = 0.66 +/- 0.13 micromol kg(-1) h(-1)) were not significantly different from the model parameters in the control experiment (KM = 0.34+/-0.06 micromol l(-1); Vmax = 0.58+/-0.07 micromol kg(-1) h(-1)) on the basis of Welch's approximate t-test. Similarly, the Michaelis-Menten model parameter estimates for Cd uptake by carp gills in the treatment experiment (KM = 0.15 +/- 0.06 micromol l(-1); Vmax = 5.14 +/- 1.07 micromol kg(-1) h(-1)) were not significantly different from the model parameters derived from the control experiment (KM =0.27 +/- 0.09 micromol l(-1); Vmax = 7.63+/-1.38 micromol kg(-1) h(-1)). This indicated that the Cd uptake rate by total carp and in carp gills in the presence of commercial humic acid followed the measured variations in Cd2+ -ion activity as predicted by the control experiment.

Sequential determination of a combined gamma/beta and pure beta-emitter by gamma and liquid scintillation counting: application to the transport of metals across fish gills.

A combined gamma scintillation/liquid scintillation technique for the sequential determination of two radioactive tracers is described. The method was developed using a combined gamma/beta-emitter (57Co) and a pure beta-emitter (45Ca). First, the 57Co radioactivity was determined by counting the samples in a gamma scintillation analyzer in the energy region 80-165 keV. Next, the samples were counted in a liquid scintillation analyzer. Only one energy region was used to count both isotopes to maximize the counting efficiencies. From the difference between quenched and unquenched beta-spectra, the counting region was set from 0 to 256 keV. The counting efficiency was related to a quench-indicating parameter (tSIE) for both nuclides by fitting a rectangular hyperbola to the quench data. By subtracting the 57Co counts from the observed counts in the total window, 45Ca dpm values were obtained. It is shown that the method presented gives reliable and consistent results. The recoveries of both isotopes are independent of the quench level in a large tSIE range, although five times more radioactivity is required for 45Ca than for 57Co to obtain accurate and reproducible results. The method has been used to study mechanisms of metal transport across biological interfaces.