Influence of temperature and surfactant on Escherichia coli inactivation in aqueous suspensions treated by moderate pulsed electric fields

This research employed a conductometric technique to estimate the inactivation kinetics of Escherichia coli cells in aqueous suspensions (1 wt.%) during simultaneous pulsed electric fields (PEF) and thermal treatments. The electric field strength was E = 5 kV/cm, the effective PEF treatment time t_PEF was within 0–0.2 s, the pulse duration t_i was 10^− 3 s, the medium temperature was 30–50 °C, and the time of thermal treatment t_T was within 0–7000 s. The damage of E. coli was accompanied by cell size decrease and release of intracellular components. The synergy between PEF and thermal treatments on E. coli inactivation was clearly present. The non-ionic surfactant Triton X-100 additionally improved its inactivation. The characteristic damage time followed the Arrhenius law within the temperature range 30–50 °C with activation energies W = 94 ± 2 kJ mol^− 1 and W = 103 ± 5 kJ mol^− 1 with and without the presence of surfactant, respectively. Relations between cell aggregation, cell ζ-potentials and presence of surfactant were analysed.     

Hydrogarnet: a host phase for Cr(VI) in chromite ore processing residue (COPR) and other high pH wastes

For understanding both the environmental behavior and developing remediation treatments for chromium ore processing residue (COPR) it is important to identify all the potentially soluble sources of Cr(VI). Hydrogarnet has been identified as a major phase in COPR and it has been previously speculated that it has a capacity to host Cr(VI). Here we provide direct evidence of this capacity by demonstrating the incorporation of Cr(VI) into laboratory synthesized hydrogarnet. Electron microscopy and energy dispersive X-ray microanalysis show incorporation of approximately 17000-22000 mg Cr(VI) kg(-1) hydrogarnet. X-ray powder diffraction data show that peak intensities are altered by chromium substitution and that chromium substituted hydrogarnets have a smaller unit cell than the pure Ca-Al end member. This is consistent with substitution of hydroxyl tetrahedra by smaller chromate tetrahedra. Electron energy loss spectroscopy confirms the tetrahedral coordination and hexavalent oxidation state of chromium in the hydrogarnets. The maximum amount of hexavalent chromium that can be introduced synthetically corresponds to a replacement of about one out of every eight hydroxyl tetrahedral per unit cell by a CrO4(2-) tetrahedra and tallies closely with the amount of chromium measured in hydrogarnets from COPR. Chromium-bearing hydrogarnet is the most abundant crystalline phase in millions of tons of COPR contaminating land around Glasgow, Scotland, and was recently identified in COPR from sites in North America. Calculations based on its abundance and its Cr(VI) content indicate that hydrogarnet can host as much as 50% of the Cr(VI) found in some COPR samples.     

Changes in endocrine profile and reproductive organs during puberty in the male marmoset monkey (Callithrix jacchus)

Data on pubertal maturation in male marmoset, a model for human reproduction, are scant and conflicting. We collected data on novel parameters to characterize puberty. Twenty-five marmoset monkeys were assigned to five age groups by weeks (wk): 21 (pre-pubertal), 43 (onset of puberty), 52 (fully pubertal), 70 (mature), and 116 (fully adult). Serum and intratesticular testosterone and pituitary bioactive chorionic gonadotropin (bioCG) were measured. Testicular development was assessed by ultrasonography, histology, and flow cytometry. Three consecutive blood samples revealed extreme fluctuations in testosterone concentrations, suggesting an erratic secretion. Age-related changes in serum testosterone and pituitary bioCG concentrations were observed. Intratesticular androgens (ITAs) showed high fluctuations within groups at all ages and were high in some animals by 21 wk. Unexpectedly, no correlation between pituitary bioCG and serum testosterone or ITAs was found, but these parameters significantly correlated with testicular weight and volume. These observations were consistent a dependence on the testis growth on bioCG. Unfortunately, the low serum levels of bioCG were not measurable in this study. At 43 wk, the animals reached puberty. At 52 wk of age, animals attained maximum body and epididymal weights and qualitatively normal spermatogenesis, but testes continued growing, reaching a maximum of all parameters at 70 wk of age, without further major changes at the age of 116 wk. It is concluded that (1) gonadal activation is evident at wk 21, (2) the male marmoset reaches the pubertal threshold around 43 wk of age, attains qualitative parameters at 52 wk, matures further to sexual maturity at 70 wk, and (3) serum testosterone and ITAs are highly variable without any identifiable correlation with pituitary bioCG.     

Developmental ability of cloned embryos from neural stem cells

The success rate is generally higher when cloning mice from embryonic stem (ES) cell nuclei than from somatic cell nuclei, suggesting that the embryonic nature or the undifferentiated state of the donor cell increases cloning efficiency. We assessed the developmental ability of cloned embryos derived from cultured neural stem cell (NSC) nuclei and compared the success rate with that of embryos cloned from other donor cells such as differentiated NSCs, cumulus cells, Sertoli cells and ES cells in the mouse. The transfer of two-cell cloned embryos derived from cultured NSC nuclei into surrogate mothers produced five live cloned mice. However, the success rate (0.5%) was higher in embryos cloned from cultured NSC nuclei than from differentiated NSCs (0%), but lower than that obtained by cloning mice from other cell nuclei (2.2-3.5%). Although the in vitro developmental potential to the two-cell stage of the cloned embryos derived from NSC nuclei (73%) was similar to that of the cloned embryos derived from other somatic cell nuclei (e.g., 85% in Sertoli cells and 75% in cumulus cells), the developmental rate to the morula-blastocyst stage was only 7%. This rate is remarkably lower than that produced from other somatic cells (e.g., 50% in Sertoli cells and 54% in cumulus cells). These results indicate that the undifferentiated state of neural cells does not enhance the cloning efficiency in mice and that the arrest point for in vitro development of cloned embryos depends on the donor cell type.     

Germin-like protein Cit s 1 and profilin Cit s 2 are major allergens in orange (Citrus sinensis) fruits

Oranges are clinically relevant allergenic foods. To date, orange allergens have not been characterized in detail. The study is aimed at analyzing the sensitization profile in orange-sensitized subjects with and without clinical allergy, and to identify orange allergens. Fifty-six sensitized subjects with self-reported reactions to orange were grouped into reactors (anaphylaxis or multiple episodes of immediate reactions and/or positive challenge tests) and non-reactors (negative open food challenge tests). Allergens were characterized by IgE immunoblotting, N-terminal sequencing, IgE-inhibition assays, and mediator release assays were performed to determine the allergenic potency of orange profilin. Of 56 subjects, 23 were classified as orange allergic showing mainly an oral allergy syndrome. Of 23 subjects classified as orange allergic, 22 were sensitized to profilin, Cit s 2. In patients with mono-sensitization to profilin in vitro histamine releases up to 75% from basophils were induced using orange extract and purified plant profilins. Of the allergic patients 78% were sensitized to germin-like protein, Cit s 1. Both allergens showed retained IgE reactivity in heat-processed orange juice. Interestingly, subjects with and without clinical allergy showed a comparable sensitization profile. Profilin and germin-like proteins are major orange allergens. The potential clinical relevance of orange profilin was indicated by its strong capacity to release histamine from basophils. However, a predominant sensitization to both allergens in subjects without symptoms also indicates a high frequency of clinically insignificant sensitization.     

Changes in numbers and kinds of bacteria during a chickpea submerged fermentation used as a leavening agent for bread production

The microflora developed during a submerged fermentation of coarsely ground chickpea (Cicer arietinum L.) in water (primary starter) and during raising a dough from wheat flour (adapted starter) was studied. In the fermenting liquid, only populations of Bacillus and Clostridium developed. Bacilli increased their loads significantly (p<0.05) during fermentation for 8-12 h and then remained constant. Clostridia developed (p<0.05) subsequently to levels of 10(7) cfu/ml at 18 h, when the pH of the fermenting liquid had decreased (p<0.05) to approximately 5.4. It also seems that the rise of the adapted starter within 2 h was caused by enzymes present in the primary starter and those liberated after cell death by the declining populations of bacilli and clostridia. The principal groups of isolates in all fermentation experiments (with chickpea seeds from five different areas) seemed to have the phenotypic characteristics of Bacillus cereus group and Clostridium perfringens. SDS-PAGE of whole-cell proteins elucidated the taxonomic position of the B. cereus group of strains as B. cereus and Bacillus thuringiensis and confirmed the phenotypic allocation of C. perfringens isolates. Strains phenotypically characterized as Bacillus licheniformis and Clostridium beijerinckii were also found to belong to these same species by SDS-PAGE. In addition, results showed that the fermenting broth was not toxic to mice when inoculated intraperitoneally and the product can thus be considered as safe for consumption.     

Update on optimized purification and characterization of natural milk allergens

Highly purified allergens namely cow's milk alpha-lactalbumin (ALA). (Bos d 4), beta-lactoglobulin (BLG) (Bos d 5) and casein (Bos d 8) and goat's milk casein were prepared from the raw milk from a single animal with a known genetic background. Consequently the natural isoforms are limited, constant and characterized. Purification included selective precipitations and chromatographical steps. Characterization of structure and allergenic activity assessment of milk allergens were carried out using physicochemical and immunochemical methods. Taken together data demonstrated the absence of impurities and of contamination by other milk allergens in each preparation. NMR and circular dichroism analyses confirmed the native conformation and proper folding of ALA and BLG and the expected absence of folding of bovine and caprine casein. Enzyme immuno assays confirmed the native conformation of BLG and the purity and immunoreactivity of all the proteins. The allergenic activity, e. g. the IgE binding capacity, of purified proteins was identical as that of those proteins when present in milk. The purified proteins also demonstrated the ability to provoke the degranulation of humanized rat basophilic leukaemia cells. All the data thus confirm the purity, identity, structural conformation and functionality of the prepared milk allergens.     

Physicochemical properties of monosodium glutamate-compounded tapioca starch exceeds those of simple heat-moisture treated starch

Monosodium glutamate (GluNa)-compounded starch was prepared by heat-moisture treating a mixture of tapioca starch and GluNa. GluNa-compounded starch exhibited a higher gelatinization temperature and reduced swelling and solubility, essentially lower hardness of the granule center, and paste viscosity than those of the heat-moisture treated tapioca starch and the untreated starch. However, its appearance, unit chain length distribution, and α-amylase digestibility were similar to those of the heat-moisture treated tapioca. It is thus concluded that GluNa compounding is useful for providing a unique type of starch that possesses a less swollen and viscous texture than that produced with simple heat-moisture treatment.     

Comparison of antimicrobial resistance of Campylobacter jejuni and Campylobacter coli isolated from humans and chicken carcasses in Poland

Campylobacter-associated gastroenteritis remains an important cause of morbidity worldwide, and some evidence suggests that poultry is an important source of this foodborne infection in humans. This study was conducted to analyze the prevalence and genetic background of resistance of 149 Campylobacter jejuni and 54 Campylobacter coli strains isolated from broiler chicken carcasses and from stool samples of infected children in Poland from 2003 through 2005. Nearly all isolates were susceptible to macrolides and aminoglycosides. The highest resistance in both human and chicken strains was observed for ciprofloxacin (more than 40%), followed by ampicillin (13 to 21%), and tetracycline (8 to 29%). Resistance to ampicillin and tetracycline rose significantly between 2003 and 2005. Slight differences in resistance between human and chicken isolates indicate that although chicken meat is not the only source of Campylobacter infection in our population, it can be involved in the transmission of drug-resistant Campylobacter strains to humans.     

Characterization of the Arcobacter contamination on Belgian pork carcasses and raw retail pork

In the present study, the occurrence of Arcobacter was assessed at four sites on 169 porcine carcasses (foreleg, chest, pelvis and ham) at different stages of slaughter and 47 pork products at retail. Carcass swab samples were enriched in Arcobacter broth containing 5-fluorouracil, amphotericine B, cefoperazone, novobiocine and trimethoprim as selective supplement. After microaerobic incubation, arcobacters were isolated using Arcobacter selective agar plates, containing the selective supplement described above. Some carcass samples and all pork samples were also examined quantitatively. All 862 isolates were identified by a species-specific m-PCR-assay and 182 isolates were further characterized by ERIC-PCR. Arcobacters were isolated from one or more sampling places on 96.4% of the carcasses, with the foreleg and the chest area as the two most contaminated sites. Furthermore, A. cryaerophilus was the most common species. Chilling decreased the number of positive carcasses, but did not eliminate all arcobacters. Direct isolation revealed that only a few carcasses were contaminated with arcobacters on foreleg and/or chest at levels higher than 10(2 )cfu/100 cm(2). Characterization demonstrated a large heterogeneity among the isolates, with ten genotypes present on more then one site per carcass. Fourteen genotypes were simultaneously present on carcasses from different herds slaughtered on the same day, which may indicate cross-contamination. Arcobacters were present in 21% of the pork samples taken at retail, but contamination levels did not exceed 100 cfu per gram. Characterization of the A. butzleri and A. cryaerophilus isolates indicated an additional contamination during processing at retail.