Treatment of Escherichia coli O157:H7 with lactic acid, neutralized electrolyzed oxidizing water and chlorine dioxide followed by growth under sub-optimal conditions of temperature, pH and modified atmosphere

The utilization of sub-lethal decontamination treatments gains more and more interest due to the increased consumers' demand for fresh, minimally processed and convenient food products. These products rely on cold chain and hurdle (combination) technology to provide microbiological safety and quality during their shelf life. To investigate the ability of surviving cells to resuscitate and grow in a food simulating environment, sub-lethal decontamination treatments were coupled with subsequent storage under sub-optimal growth conditions. For this purpose chlorine dioxide (ClO₂) and neutralized electrolyzed oxidizing water (NEW)-treated cultures of Escherichia coli O157:H7 were inoculated in TSB-YE of pH 5.8 and a_w 0.99, and stored at 10 °C, 12.5 °C and 15 °C, under four different atmospheres (0%, 30% and 60% CO₂ balanced with N₂, and air). Due to the severity of injury, lactic acid-treated cells were inoculated in TSB-YE pH 7.0. Data obtained reveal that the fraction of sub-lethally injured E. coli O157:H7 undergoes an additional inhibitory effect during the storage period under of sub-optimal conditions. Observed extension in the lag growth phase was a direct consequence prior sub-lethal injury. The effects of liquid ClO₂ and NEW were less pronounced in comparison to lactic acid. The current study signifies the potential utilization of appropriate combination of different extrinsic and intrinsic factors in the elimination or growth inhibition of food-borne pathogens.     

The mechanical properties of densified VTC wood relevant for structural composites

The mechanical properties of densified wood relevant for structural composites were studied. Low density hybrid poplar (Populus deltoides × Populus trichocarpa) was densified using the viscoelastic thermal compression (VTC) process to three different degrees of densification (63, 98, and 132%). The modulus of rupture (MOR) and the modulus of elasticity (MOE) of the control (undensified) wood and of the VTC wood were determined. The bonding performance of the control and VTC wood, using two phenol-formaldehyde (PF) adhesives, was studied. Four different 3-layer composites were also prepared from undensified and VTC wood, and tested in four-point bending. The results showed that the bending properties of the VTC wood (MOR and MOE) were significantly improved due to the increased density. The bonding performance of VTC wood with PF adhesives was comparable with or better than in the case of the control wood. Increased density of the face layers in the 3-layer VTC composites was advantageous for their mechanical performance.     

Growth of Escherichia coli O157:H7 and Listeria monocytogenes with prior resistance to intense pulsed light and lactic acid

Previous study showed that repetitive mild decontamination treatments with intense light pulses (ILP) and lactic acid (LA) can induce increased resistance in surviving pathogenic cells. Research has evaluated the potential of increased resistance to enhance the persistence of resistant variants of Listeria monocytogenes and Escherichia coli O157:H7 under suboptimal growth conditions. Growth of resistant variants and parental strains was determined by optical density (OD) measurements in nutrient broths with different pH values and NaCl concentration, at low temperature. The real lag phase was calculated, and results indicated that intense light pulses (ILP) resistant variants needed longer time to initiate growth compared to their parental strains, for both L. monocytogenes and E. coli O157:H7 when incubated at 7 °C and 10 °C, respectively. These selected variants were of the similar resistance towards heat and low pH (no cross-tolerance). Nevertheless, lactic acid (LA) resistant variant of L. monocytogenes was cross-protected when exposed to low pH, but not when treated with heat.     

Detection of <Emphasis Type="Italic">Clostridium botulinum</Emphasis> neurotoxins A and B in milk by ELISA and immuno-PCR at higher sensitivity than mouse bio-assay

Using commercially available antibodies and toxoids as templates, an ELISA, immuno-quantitative PCR (iqPCR), and multiplex immuno-PCR (iPCR) were developed for detection of Clostridium botulinum neurotoxins A and B. The obtained sensitivities for ELISA ranged from 1 ng/ml in PBS + 1% BSA to 15 and 10 ng/ml in skimmed milk for botulinum neurotoxins (BoNT)/A and BoNT/B, respectively. In semi-fat milk, the limit of detection (LOD) for both toxoids was 30 ng/ml. Quantitative immuno-PCR (iqPCR) had an LOD of 4.5–9 pg/reaction for BoNT/A in both PBS and semi-fat milk, while this was 18.5–37 pg/reaction for BoNT/B in PBS + 1% BSA and semi-fat milk, respectively. The sensitivities of ELISA and iqPCR were improved to 0.5 ng/ml and 3.75 pg/ml (0.2 pg/reaction) in semi-fat milk, respectively, when toxoid of BoNT/A was substituted with actual toxin. Multiplex iPCR with both toxoids run in the same reaction was able to distinguish presence/absence of tested BoNT/A and BoNT/B at 25 pg/reaction. The tested system offers a realistic alternative with much better sensitivity to the standard mouse assay.     

Performance of Drying Technologies to Ensure Microbial Safety of Dried Fruits and Vegetables

Dried fruits, vegetables, herbs, and spices are produced in and sourced from many countries worldwide, but they have been increasingly reported to be involved in outbreaks and alerts due to the presence of foodborne pathogens such as Salmonella. These dried products are mainly produced by solar drying and conventional air drying, but a wide range of drying technologies are available. From a technological point of view the general trend is to optimize and standardize the drying process to ensure high‐quality products to be offered. Drying technologies are mainly evaluated for their performance to reduce water activity at low energy cost while maintaining good sensorial quality of the dried product. However, as low water activity foods are increasingly recognized to support microbial survival and dried products are often consumed as they are, or are used as ingredients in many ready‐to‐eat foods, there is increasing attention to the microbiological quality and safety aspects of these products. This review presents traditional and emerging technologies to dry fruits, vegetables, herbs, and spices and discusses their potential to inactivate bacteria and viruses throughout the drying process. Overall, the microbial inactivation effect of the presented technologies has not yet been thoroughly assessed, even for traditional methods like solar drying, conventional air drying, or freeze‐drying. Emerging technologies such as dielectric (assisted) drying and low‐pressure superheated steam drying have been shown to reduce microbial populations; however, the number of studies is still low. Very few studies have focused on viral inactivation during drying processes.     

The Human Milk Microbiota Produces Potential Therapeutic Biomolecules and Shapes the Intestinal Microbiota of Infants

Human milk not only provides a perfect balance of nutrients to meet all the needs of the infant in the first months of life but also contains a variety of bacteria that play a key role in tailoring the neonatal faecal microbiome. Microbiome analysis of human milk and infant faeces from mother-breastfed infant pairs was performed by sequencing the V1–V3 region of the 16S rRNA gene using the Illumina MiSeq platform. According to the results, there is a connection in the composition of the microbiome in each mother-breastfed infant pair, supporting the hypothesis that the infant’s gut is colonised with bacteria from human milk. MiSeq sequencing also revealed high biodiversity of the human milk microbiome and the infant faecal microbiome, whose composition changes during lactation and infant development, respectively. A total of 28 genetically distinct strains were selected by hierarchical cluster analysis of RAPD-PCR (Random Amplified Polymorphic DNA-Polymerase Chain Reaction) electrophoresis profiles of 100 strains isolated from human milk and identified by 16S RNA sequencing. Since certain cellular molecules may support their use as probiotics, the next focus was to detect (S)-layer proteins, bacteriocins and exopolysaccharides (EPSs) that have potential as therapeutic biomolecules. SDS-PAGE (Sodium Dodecyl-Sulfate Polyacrylamide Gel Electrophoresis) coupled with LC-MS (liquid chromatography-mass spectrometry) analysis revealed that four Levilactobacillus brevis strains expressed S-layer proteins, which were identified for the first time in strains isolated from human milk. The potential biosynthesis of plantaricin was detected in six Lactiplantibacillus plantarum strains by PCR analysis and in vitro antibacterial studies. ¹H NMR (Proton Nuclear Magnetic Resonance) analysis confirmed EPS production in only one strain, Limosilactobacillus fermentum MC1. The overall microbiome analysis suggests that human milk contributes to the establishment of the intestinal microbiota of infants. In addition, it is a promising source of novel Lactobacillus strains expressing specific functional biomolecules.     

Optimization of industrial‐scale deodorization of high‐oleic sunflower oil via response surface methodology

Optimization of industrial‐scale deodorization of high‐oleic sunflower oil (HOSO) via response surface methodology is presented in this study. The results of an experimental program conducted on an industrial‐scale deodorizer were analyzed statistically. Predictive models were derived for each of the oil quality indicators (QI) in dependence on the studied variable deodorization process parameters. The deodorization behavior of some minor components was analyzed on a pilot‐scale deodorizer. For comparison, a similar experimental program was also performed on the laboratory‐scale. The results of this study demonstrate that optimization of the deodorization process requires a suitable compromise between often mutually opposing demands dictated by different oil QI. The production of HOSO with top‐quality organoleptic and nutritional values (high tocopherol and phytosterol contents and low free and trans fatty acid contents) and high oxidative stability demands deodorization temperatures in the range between 220 and 235 °C and a total sparge steam above 2.0% (wt/wt in oil). The response surface methodology provides the tools needed to identify the optimum deodorization process conditions. However, the laboratory‐scale experiments, while showing similar response characteristics of QI in dependence on the process parameters and thus helpful as a guide, are of limited value in the optimization of an industrial‐scale operation.     

Rheological examination of temperature dependence of conventional and polymer‐modified road bitumens

Rheological characterization of two types of road bitumens, conventional and polymer‐modified, has been examined in the temperature range between 20°C and 140°C. Tests were carried out before and after ageing following a thin‐film oven test. Polymer‐modified bitumens exhibit non‐Newtonian behaviour up to the 120°C due to a complex secondary structure formed by added polymers. For conventional bitumens, Newtonian behaviour was observed above 60°C. Special attention was paid to measurements and to analysis of the dynamic data of oscillatory shear. The mechanical spectra in a wide frequency range have been obtained using the WLF time‐temperature superposition principle. The analysis of viscoelastic data clearly showed the differences between the two types of bitumen. Conventional bitumens were more sensitive to temperature and to the ageing effects. For polymer modified bitumens, the elastic contribution to viscoelastic response was more pronounced, and independent of temperature and ageing.