Carmen I. Moraru

Effect of Micro- and Nanoscale Topography on the Adhesion of Bacterial Cells to Solid Surfaces

ABSTRACT Attachment and biofilm formation by bacterial pathogens on surfaces in natural, industrial, and hospital settings lead to infections and illnesses and even death. Minimizing bacterial attachment to surfaces using controlled topography could reduce the spreading of pathogens and, thus, the incidence of illnesses and subsequent human and financial losses. In this context, the attachment of key microorganisms, including Escherichia coli, Listeria innocua, and Pseudomonas fluorescens, to silica and alumina surfaces with micron and nanoscale topography was investigated. The results suggest that orientation of the attached cells occurs preferentially such as to maximize their contact area with the surface. Moreover, the bacterial cells exhibited different morphologies, including different number and size of cellular appendages, depending on the topographical details of the surface to which they attached. This suggests that bacteria may utilize different mechanisms of attachment in response to surface topography. These results are important for the design of novel microbe-repellant materials.

Inactivation of Escherichia coli ATCC 25922 and Escherichia coli O157:H7 in apple juice and apple cider, using pulsed light treatment.

The main objective of this work was to evaluate the effectiveness of pulsed light (PL) treatment for the inactivation of Escherichia coli in liquids with different levels of clarity. Nonpathogenic E. coli ATCC 25922 and pathogenic E. coli O157: H7 were used as challenge organisms. Butterfields phosphate buffer (BPB), tryptic soy broth (TSB), apple juice, and apple cider were used as substrates. The inoculated liquids were placed in a thin layer (1.3 mm) into glass chambers (23 by 53 by 11 mm) and exposed to PL doses of up to 13.1 J/cm2. PL treatments were performed in a Xenon RS-3000C PL unit, both in static mode and under turbulence. Survivors were determined by standard plate counting or the most-probable-number technique. For static treatments, reduction levels exceeding 8.5 log were obtained in BPB for all strains and reduction levels of about 3.5 log were obtained in TSB. For apple juice, inactivation levels of 2.66 +/- 0.10 log were obtained for E. coli ATCC 25922 and 2.52 +/- 0.19 log for E. coli O157:H7. In cider, inactivation levels of 2.32 +/- 0.16 log and 3.22 +/- 0.29 log were obtained for the nonpathogenic and pathogenic strains, respectively. Inactivation kinetics was characterized using the Weibull model. Turbulent treatments resulted in 5.76 +/- 0.06 log reduction in cider and 7.15 +/- 0.22 log reduction in juice, which satisfies the U.S. Food and Drug Administration requirement of 5-log reduction of E. coli. These results show promise for the use of PL for the effective reduction of E. coli in apple juice and cider.

Reduction of Listeria on ready-to-eat sausages after exposure to a combination of pulsed light and nisin.

The risk of listeriosis associated with ready-to-eat foods is a major concern in the United States. Pulsed light (PL) treatment has been effective for killing Listeria. The possibility of enhancing the antilisterial capability of PL treatment by combining PL with an additional hurdle, the natural antimicrobial nisin, was explored in this study. First, the ability of Listeria innocua to mimic the response of Listeria monocytogenes to PL treatment was demonstrated. Subsequently, a series of inoculation studies was performed in which canned sausages were surface inoculated with L. innocua as a surrogate for L. monocytogenes and then treated with a commercial preparation of nisin (Nisaplin), PL, or a combination of the two treatments. The application of a Nisaplin dip alone resulted in an immediate reduction of L. innocua by 2.35 +/- 0.09 log CFU. PL reduced L. innocua by 1.37 +/- 0.30 log CFU after exposure to 9.4 J/cm2. A total reduction of 4.03 +/- 0.15 log CFU was recorded after the combined treatment of Nisaplin and PL for 48 h at 4 degrees C. The long-term survival of L. innocua was evaluated on sausages stored at 4 degrees C. Treatment with Nisaplin and PL resulted in a 4- to 5-log reduction for two replicate studies. The combination treatment resulted in no significant microbial growth during 28 and 48 days of refrigerated storage in the first and second replicates, respectively. These results suggest that this combination treatment can be used as an effective antilisterial step in the production of ready-to-eat foods.

Effect of Xanthan Gum and CMC on the Structure and Texture of Corn Flour Pellets Expanded by Microwave Heating

ABSTRACT Third-generation snacks obtained by microwave expansion of glassy cereal pellets are often hard and nonuniform and need to be improved to become acceptable to the consumer. Gums such as carboxymethyl cellulose (CMC) and xanthan gum (XG) have the ability to improve the volume, structure, and texture of expanded cereal products, due to their effects on moisture retention and rheological properties. This study investigates the effect of 1% addition of CMC and XG on the structural and mechanical properties of samples obtained by microwave expansion of glassy corn pellets. Unexpanded, glassy pellets were obtained by extrusion and subsequent equilibration at aw = 0.53 at room temperature. The equilibrated pellets were expanded by microwave heating. The addition of gums significantly improved the shape, structural and textural uniformity of the microwave-expanded samples. These effects were attributed to the contribution of gums to the rheology and moisture sorption capacity of the matrix. It is our hyp...

Inactivation kinetics and factors of variability in the pulsed light treatment of Listeria innocua cells.

Pulsed light (PL) treatment can effectively reduce microbial populations in clear substrates and on surfaces, but its effectiveness varies as a function of substrate or treatment-related factors. For PL to be successfully adopted by the food industry, all factors of influence, as well as the inactivation kinetics for the microorganisms of concern, must be elucidated. In this study, the inactivation kinetics of Listeria innocua and the effect of inoculum size on PL inactivation were investigated. Stainless steel coupons (50.8 by 101.6 mm) of defined surface properties and transparent glass chamber slides (25.4 by 50.8 by 10 mm) were each inoculated with 1 ml of aqueous suspensions of L. innocua containing inoculum populations of up to 10(9) CFU. The thickness of the liquid layer in the glass slides was 1.16 mm. The inoculated substrates were exposed to PL treatment of up to 17 J/cm2 in a static PL chamber equipped with a pulsed Xenon lamp. Survivors were recovered and enumerated by both standard plate counting and most-probable-number procedures. The data indicated that in clear liquids, PL resulted in more than a 6-log reduction of L. innocua after a 12-J/cm2 treatment, regardless of the initial inoculum size. For the stainless steel surfaces, less than a 4-log reduction after a 12-J/cm2 treatment and a noticeable effect of substrate characteristics and inoculum size on inactivation were observed. The survivor curves showed pronounced tailing for all substrates used in the study. The Weibull model accurately predicted the survivor ratios for the PL treatment of L. innocua in clear liquids, with a shape and scale parameter of 0.33 and 3.01, respectively. The Weibull model resulted in significant overestimation of PL effectiveness for the stainless steel substrates, where the influence of various substrate properties and inoculum level on inactivation was significant.

Factors That Influence the Microwave Expansion of Glassy Amylopectin Extrudates

ABSTRACT The microwave expansion of glassy, unexpanded amylopectin pellets was studied. Amylopectin was extruded at three levels of specific mechanical energy (483, 809, and 846 kJ/kg), and 35–40% moisture content, without expansion at the die. Glassy pellets were obtained by drying and equilibrating the extrudates at five water activities (aw 0, 0.11, 0.33, 0.67, and 0.75). The pellets were characterized by measuring volume, porosity, and moisture content. The pellets were then expanded in a constant power microwave oven to determine the degree of expansion. When subjected to microwave heating, regardless of extrusion condition and initial aw, the pellets expanded from the center where the highest temperature was recorded and then expansion advanced in the whole volume. Maximum expansion was reached after 30 sec of heating, after which samples started to burn from the center. Samples simultaneously expanded and lost moisture, both processes being faster and more intense for pellets of higher initial aw. ...

Effect of spectral range in surface inactivation of Listeria innocua using broad-spectrum pulsed light.

Pulsed light (PL) treatment is an alternative to traditional thermal treatment that has the potential to achieve several log-cycle reductions in the concentration of microorganisms. One issue that is still debated is related to what specifically causes cell death after PL treatments. The main objective of this work was to elucidate which portions of the PL range are responsible for bacterial inactivation. Stainless steel coupons with controlled surface properties were inoculated with a known concentration of Listeria innocua in the stationary growth phase and treated with 1 to 12 pulses of light at a pulse rate of 3 pulses per s and a pulse width of 360 micros. The effects of the full spectrum (lambda = 180 to 1,100 nm) were compared with the effects obtained when only certain regions of UV, visible, and near-infrared light were used. The effectiveness of the treatments was determined in parallel by the standard plate count and most-probable-number techniques. At a fluence of about 6 J/cm(2), the full-spectrum PL treatment resulted in a 4.08-log reduction of L. innocua on a Mill finish surface, the removal of lambda < 200 nm diminished the reduction to only 1.64 log, and total elimination of UV light resulted in no lethal effects on L. innocua. Overwhelmingly, the portions of the PL spectrum responsible for bacterial death are the UV-B and UV-C spectral ranges (X < 300 nm), with some death taking place during exposure to UV-A radiation (300 < lambda < 400 nm) and no observable death upon exposure to visible and near-infrared light (lambda > 400 nm). This work provides additional supporting evidence that cell death in PL treatment is due to exposure to UV light. Additionally, it was shown that even a minor modification of the light path or the UV light spectrum in PL treatments can have a significant negative impact on the treatment intensity and effectiveness.

Effect of solvent and temperature on the size distribution of casein micelles measured by dynamic light scattering

The objectives of this study were to investigate the effect of the solvent on the accuracy of casein micelle particle size determination by dynamic light scattering (DLS) at different temperatures and to establish a clear protocol for these measurements. Dynamic light scattering analyses were performed at 6, 20, and 50 degrees C using a 90Plus Nanoparticle Size Analyzer (Brookhaven Instruments, Holtsville, NY). Raw and pasteurized skim milk were used as sources of casein micelles. Simulated milk ultrafiltrate, ultrafiltered water, and permeate obtained by ultrafiltration of skim milk using a 10-kDa cutoff membrane were used as solvents. The pH, ionic concentration, refractive index, and viscosity of all solvents were determined. The solvents were evaluated by DLS to ensure that they did not have a significant influence on the results of the particle size measurements. Experimental protocols were developed for accurate measurement of particle sizes in all solvents and experimental conditions. All measurements had good reproducibility, with coefficients of variation below 5%. Both the solvent and the temperature had a significant effect on the measured effective diameter of the casein micelles. When ultrafiltered permeate was used as a solvent, the particle size and polydispersity of casein micelles decreased as temperature increased. The effective diameter of casein micelles from raw skim milk diluted with ultrafiltered permeate was 176.4 +/- 5.3 nm at 6 degrees C, 177.4 +/- 1.9 nm at 20 degrees C, and 137.3 +/- 2.7 nm at 50 degrees C. This trend was justified by the increased strength of hydrophobic bonds with increasing temperature. Overall, the results of this study suggest that the most suitable solvent for the DLS analyses of casein micelles was casein-depleted ultrafiltered permeate. Dilution with water led to micelle dissociation, which significantly affected the DLS measurements, especially at 6 and 20 degrees C. Simulated milk ultrafiltrate seemed to give accurate results only at 20 degrees C. Results obtained in simulated milk ultrafiltrate at 6 degrees C could not be explained based on the known effects of temperature on the casein micelle, whereas at 50 degrees C, precipitation of amorphous calcium phosphate affected the DLS measurement.

Effect of Specific Mechanical Energy on Properties of Extruded Protein-Starch Mixtures

ABSTRACT The effect of the specific mechanical energy (SME) during extrusion of a protein-starch mixture was studied by analyzing the glass transition temperature (Tg) and starch gelatinization. We found that the SME values of 344 to 2108 kJ/kg did not significantly change the Tg of the product. To explain the insensitivity of Tg to SME in spite of reported fragmentation taking place during extrusion, we studied the effect of the molecular weight (MW) on Tg in a model system consisting of dextrans of varying molecular weights. We found that the effect of the molecular weight on the Tg reached a plateau at 6.7 × 104. Because the reported size of the fragments created during the extrusion process is larger than this, we were able to explain the apparent insensitivity of Tg to SME in the protein-carbohydrate matrix studied. However, we found that starch gelatinization varied with SME, the degree of gelatinization being higher for systems exposed to higher SME.

Bacterial attachment and biofilm formation on surfaces are reduced by small-diameter nanoscale pores: how small is small enough?

Background/Objectives:Prevention of biofilm formation by bacteria is of critical importance to areas that directly affect human health and life including medicine, dentistry, food processing and water treatment. This work showcases an effective and affordable solution for reducing attachment and biofilm formation by several pathogenic bacteria commonly associated with foodborne illnesses and medical infections.Methods:Our approach exploits anodisation to create alumina surfaces with cylindrical nanopores with diameters ranging from 15 to 100 nm, perpendicular to the surface. The anodic surfaces were evaluated for attachment by Escherichia coli, Listeria monocytogenes, Staphylococcus aureus and Staphylococcus epidermidis. Cell–surface interaction forces were calculated and related to attachment.Results:We found that anodic alumina surfaces with pore diameters of 15 and 25 nm were able to effectively minimise bacterial attachment or biofilm formation by all the microorganisms tested. Using a predictive physicochemical approach on the basis of the extended Derjaguin and Landau, Verwey and Overbeek (XDLVO) theory, we attributed the observed effects largely to the repulsive forces, primarily electrostatic and acid–base forces, which were greatly enhanced by the large surface area originating from the high density, small-diameter pores. We also demonstrate how this predictive approach could be used to optimise different elements of surface topography, particularly pore diameter and density, for further enhancing the observed bacteria-repelling effects.Conclusions:We demonstrate that anodic nanoporous surfaces can effectively reduce bacterial attachment. These findings are expected to have immediate, far-reaching implications and commercial applications, primarily in health care and the food industry.