Detection of verocytotoxin-producing Escherichia coli serogroups 0157 and 026 in the cecal content and lymphatic tissue of cattle at slaughter in Italy

Verocytotoxin-producing Escherichia coli (VTEC) has emerged as a foodborne pathogen that can cause severe and potentially fatal illnesses, such as hemorrhagic colitis or the hemolytic uremic syndrome. In this study, 182 cattle at slaughter (119 dairy cows and 63 feedlot cattle) were randomly selected and tested for the presence of VTEC serogroups O26, O103, O111, O145, and O157 in their cecal content and lymphatic tissue (tonsils or mesenteric lymph nodes). A total of 364 samples were evaluated with an immunomagnetic separation technique followed by slide agglutination. Presumptive VTEC 026, O103, O111, O145, and O157 isolates were tested by Vero cell assay for verocytotoxin production and by multiplex PCR assay for the detection of vtxl, vtx2, eae, and E-hlyA genes. VTEC O157 was detected in 6 (3.3%) of 182 animals, and VTEC 026 was detected in 1 (0.5%) of 182 animals. No VTEC O103, VTEC O111, or VTEC O145 isolates were found in cattle feces, but one VTEC O91:H- vtx2+, eae-, E-hlyA+ strain nonspecifically cross-reacted with the VTEC O103 type. The prevalence of VTEC O157 in the lymphatic tissue of cattle was 1.1% in both tonsils (1 of 93 samples) and mesenteric lymph nodes (1 of 89 samples). Lymphatic tissue contamination was observed only in VTEC O157 intestinal carriers; two (33.3%) of six fecal carriers were simultaneously VTEC O157 lymphatic carriers. This finding suggests that VTEC O157 contamination of meat does not necessarily come from feces or the environment. No other VTEC serogroups were detected in the lymphatic tissue of slaughtered cattle.     

Antimicrobial performance of potassium sorbate supported in tapioca starch edible films

The release and antimicrobial activity of potassium sorbate (KS) supported in tapioca starch–glycerol edible films prepared by different gelatinization/drying techniques against Zygosaccharomyces bailii was studied. Antimicrobial release in liquid media of different pHs (3.0–6.0) could be approximated to a pseudo first-order kinetic model and was almost accomplished after 30 min. Filmmaking method involving slow gelatinization and drying rate resulted in the highest fraction of KS released at equilibrium. Rate constant was higher when pH of the receiving media was 4.5 and fast gelatinization/fast drying had been used. The effectiveness of the preservative released for controlling the microbial growth depended on the pH of the receiving media, being higher at pH 3.0. No effect of filmmaking method was observed. In relation to film effectiveness as a barrier to contamination, it was observed that the preservative was available to prevent an external Z. bailii contamination and also to control yeast growth in an acidified (pH 4.5) high water activity (a _w = 0.980) semisolid product.     

A systematic method for predetermined scanning electron microscope analysis in dental science

Scanning electron microscope evaluation could be criticized if the method adopted to correct for bias is not specified in the study design. Observers can draw conclusions from images unconsciously chosen to best support their research hypotheses, impairing the basic research principle of operator's impartiality. In this study, a systematic observation method has been described and verified for repeatability. The number and the observation points on a certain specimen have been predetermined using a scheme along with observation rules previously established in the research protocol. When our instrument is used at an operating magnification between 500× and 1,000× (corresponding to a frame of 250×190 µ and 120×90 µ, respectively), the method allowed 100% repeatable observation frames, with linear frame errors in finding an observation point of 12.5% in length and 16.8% in height. With modifications to accommodate research objective and statistical requirements, the method could be applied to many SEM observation study. SCANNING 32: 97–103, 2010. © 2010 Wiley Periodicals, Inc.     

Immunochemical or fluorescent labeling of vesicular subcellular fractions for microscopy imaging

We describe a procedure for the labeling of membranous vesicular purified subcellular fractions, to image them, typically by confocal laser scanning microscopy. Being intracellular organelles, these fractions, once purified cannot be attached to glass slides as for cells. Fractions are labeled “in batch” without prior embedding or freezing. Each labeling step performed by passages of resuspension/centrifugation is followed by washings. Then samples are dispersed on the glass slides. Mammalian retinal rod outer segment disks, intact brain stem myelin vesicles, and brain synaptosomes were chosen, as these subcellular fractions can be purified by well established procedures. These fractions were immunolabeled with specific antibodies. Moreover, by the earlier procedure, we show that the mitochondrial vital membrane potential probe MitoTracker Deep Red 633 stains myelin vesicles and rod disks before fixation, consistently with our previous reports of a respiring capacity of these membranes. Therefore, the technique seems adequate to become an instrument to study the structure and the function of these and other subcellular fractions. Microsc. Res. Tech. 73:1086–1090, 2010. © 2010 Wiley‐Liss, Inc.     

Influence of processing on the levels of amines and proline and on the physico-chemical characteristics of concentrated orange juice

The influence of processing on the levels of bioactive amines and proline as well as on some physico-chemical characteristics of concentrated orange juice was investigated. Samples were collected at seven points of a production line on three non-consecutive days, and analysed for the levels of 12 bioactive amines, proline, and reducing and non-reducing sugars, and for some physico-chemical characteristics. Among the amines analysed, only putrescine, spermidine and synephrine were found in the samples at all processing stages. Significant differences were observed for spermidine and total amine levels throughout processing. Synephrine and putrescine were resistant to processing. Proline levels changed significantly throughout processing, as did levels of reducing and non-reducing sugars. The pH varied from 3.59 to 3.72, acidity from 0.873 to 0.918 g citric acid/100 mL, and density from 1.0378 to 1.0970 g/mL. Among these parameters, pH, acidity and density were significantly affected by processing.     

Effect of phosphate binders on oxidative stress and inflammation markers in hemodialysis patients

It has been suggested that phosphate binders may reduce the inflammatory state of hemodialysis (HD) patients. However, it is not clear whether it has any effect on oxidative stress. The objective of this study was to evaluate the effect of sevelamer hydrochloride (SH) and calcium acetate (CA) on oxidative stress and inflammation markers in HD patients. Hemodialysis patients were randomly assigned to therapy with SH (n=17) or CA (n=14) for 1 year. Before the initiation of therapy (baseline) and at 12 months, we measured in vitro reactive oxygen species (ROS) production by stimulated and unstimulated polymorphonuclear neutrophils and serum levels of tumor necrosis factor α, interleukin-10, C-reactive protein, and albumin. There was a significant reduction of spontaneous ROS production in both groups after 12 months of therapy. There was a significant decrease of Staphylococcus aureus stimulated ROS production in the SH group. There was a significant increase in albumin serum levels only in the SH group. In the SH group, there was also a decrease in the serum levels of tumor necrosis factor α and C-reactive protein. Our results suggest that compared with CA treatment, SH may lead to a reduction in oxidative stress and inflammation. Therefore, it is possible that phosphate binders exert pleiotropic effects on oxidative stress and inflammation, which could contribute toward decreasing endothelial injury in patients in HD.     

A Novel Function of the Triterpene Squalene in a Tritrophic System

Changes in apple leaf chemistry after infestation by leafminers and their effect on both host location and host habitat location of the generalist parasitoid Pholetesor bicolor were investigated. Chemical analysis of leaf solvent extracts from healthy and leafminer-damaged leaves revealed that herbivory increased the amount of the triterpene squalene (C₃₀H₅₀), whereas quantities of all other identified compounds were similar in both plant treatments. To assess the response of parasitoids to host location cues, contact bioassays were conducted with naïve females. Results showed that parasitoids performed a characteristic ovipositional probing more often on the mine-damaged than on the healthy leaf. This behavior was triggered by a hexane extract of the mine-damaged leaf, but not by a healthy leaf extract. A synthetic mixture of the compounds identified in the extract triggered a similar response. A mixture devoid of squalene was not active, whereas squalene alone elicited the probing behavior. To assess the use of the identified compounds in habitat location, Y-tube olfactometer experiments were conducted with naïve and experienced females. Results showed that squalene is not involved in habitat location and has no priming effect on P. bicolor. While other triterpenes are known to mediate habitat location of parasitoids, this is the first report in which a plant triterpene is shown to mediate host location of a parasitoid. The biological and ecological functions of squalene on all three trophic levels are discussed.     

Comparison of electromyographic pattern of sensory experts and untrained subjects during chewing of Mahon cheese

Use of electromyography (EMG) to monitor mastication is a relatively new concept in assessing food physical and sensory properties. Although expert assessment of cheese characteristics is widely used, the effect of training in sensory analysis on mastication patterns, as assessed using EMG is not well known. Nine samples of the same Mahon cheese (60 days ripening) were given to 24 subjects (8 experts, 16 untrained) and EMG recorded for each chewing sequence. Three samples were tested in a single session by each subject, and three sessions carried out on different days. EMG was recorded from four masticatory muscles for each subject. From EMG records the following was extracted: number of chews, chewing time, mean and maximum voltage of EMG bursts (i.e. chews) across chewing sequence, chewing work and chewing rate. No gender bias was found for the EMG parameters considered, therefore, as regards gender, each group was considered to be homogeneous. Variability within-subjects across samples was greater for experts than untrained subjects. Significant differences in chewing time, chewing work and chewing rate were found between the expert and untrained groups. Data analysis of the three sessions showed an influence of cognitive constructs, mediating states, on the chewing process. The experts were found to be goal driven as to their mastication process. Experts showed no significant differences between sessions, untrained subjects were found to vary their EMG output in successive sessions for number of chews, chewing time, mean voltage, and chewing work.