Role of yeasts in table olive production

Table olives are a traditional fermented vegetable of the Mediterranean countries, but their production and consumption are now spread all around the world. Yeasts can play a double role in this food. They are present throughout the fermentative process and it is generally accepted that they can produce compounds with important organoleptic attributes determining the quality and flavour of the final product. However, yeasts can also be spoilage microorganisms in olive fermentation/storage and packing causing gas pockets, swollen containers, cloudy brines and off-flavours and off-odours. Candida boidinii, Debaryomyces hansenii, Pichia anomala, P. membranifaciens, Rhodotorula glutinis and Saccharomyces cerevisiae are species isolated with a high frequency from olive processes. Scarce information is still available about the ecology, biochemistry and molecular biology of these important microorganisms in table olives. A wider knowledge about these aspects could facilitate the possible application of yeasts as a starter culture, due to their ability to produce aromatic compounds, antioxidants, enzymes, and improve the growth of lactic acid bacteria.     

Inactivation of Oxidative Enzymes by High-Intensity Pulsed Electric Field for Retention of Color in Carrot Juice

The effects of high-intensity pulsed electric fields (HIPEF) on oxidative enzymes and color of fresh carrot juice were studied. A response surface methodology (RSM) was used to evaluate the effect of pulse polarity (mono or bipolar mode), pulse width (from 1 to 7 μs), and pulse frequency (from 50 to 250 Hz) on color and peroxidase (POD) inactivation of carrot juice treated by HIPEF. The total treatment time and the electric field strength were set at 1,000 μs and 35 kV/cm, respectively, at a temperature below 35°C. The physicochemical characteristics of carrot juice were measured. There was a linear relationship between electrical conductivity and temperature of the carrot juice. The results showed that HIPEF-treated carrot juice at 35 kV/cm for 1,000 μs applying 6 μs pulse width at 200 Hz in bipolar mode led to 73.0% inactivation of POD. The color coordinates did not change significantly. Therefore, HIPEF was effective in POD inactivation and carrot juice color preservation.     

Influence of grape variety and their phenolic composition on vasorelaxing activity of young red wines

Red wine has been reported to exert beneficial effects in preventing cardiovascular diseases probably due to their polyphenols constituents. The vasorelaxant capacity and phenolic composition of four monovarietal young red wines Merlot, Tempranillo, Garnacha and Cabernet-Sauvignon obtained from grapes from 2004 vintage, cultivated under the same climatic and agricultural conditions, wine-making technology and storage conditions, were determined using vascular reactivity assay in rat aortic and high-performance liquid chromatography, respectively. Primary cultures of vascular smooth muscle cells from Wistar rats were also used to evaluate their survival prevention activity as well. Every studied wine present vasorelaxation effect, but the higher value corresponds to the analyzed Merlot wine, especially rich in phenolic compounds, mainly catechins and oligomeric proanthocyanidins and anthocyanin glycosides. This study further demonstrated previous investigations about the phenolic composition of wines relation with their vasorelaxation activity.     

Toxicity increases in ice containing monochlorophenols upon photolysis: environmental consequences

The toxic effects of photoproducts formed upon the photolysis of 2- and 4-chlorophenol (CP) frozen solutions in polycrystalline ice phase were determined with a bacterial luminescence test (Vibrio fisheri), and in vitro biomarker assay for dioxin-like effects (inductions of AhR-dependent luciferase in H4IIE-luc cells) and compared to the toxic effects of products of the same photoreaction in aquatic phase. Coupling photoproducts formed in ice samples (3'-chlorobiphenyl-2,4'-diol and 3-chlorobiphenyl-2,2'-diol from 2-CP photolysis and 5-chlorobiphenyl-2,4'-diol from 4-CP photolysis) were found to be more toxic to V. fisheri than parent CPs and elicited significant inductions of dioxin-like effects (the effective concentrations EC50 approximately 3 x 10(-5) mol L(-1) corresponded to known weaker ligands of AhR, such as nonplanar polychlorinated biphenyls or polycyclic aromatic hydrocarbons). To complete the picture, a photoproduct formed from 4-CP (5-chlorobiphenyl-2,4'-diol) was synthesized, and a detailed toxicity assessment with purified compound confirmed the results obtained with irradiated samples. Our findings support a recently proposed model according to which solar radiation can trigger the formation of new types of organic pollutants in polar ice or tropospheric ice cloud particles, presenting possibly greater risk to the environment than the parent compounds.     

Angiotensin I-converting enzyme inhibitory activity of peptides derived from egg white proteins by enzymatic hydrolysis

The hydrolysis of crude egg white with pepsin, trypsin, and chymotrypsin produced peptides with angiotensin-converting enzyme (ACE) inhibitory properties. These peptides were mainly derived from the proteolysis of ovalbumin. The most active hydrolysates were obtained after treatment with pepsin (50% inhibitory concentration [IC50], 55.3 microg/ml), with the fraction having a molecular mass lower than 3,000 Da giving the highest ACE inhibitory activity (IC50, 34.5 microg/ml). Nine subfractions were collected from the fraction with a molecular mass lower than 3,000 Da using semipreparative reversed-phase high-performance liquid chromatography. Considerable ACE inhibitory activity (IC50 < 40 microg/ml) was found in three of them. These subfractions were analyzed by reversed-phase high-performance liquid chromatography-tandem mass spectrometry, and 14 peptides were identified. These sequences were synthesized, and their ACE inhibitory activities were measured. Among the identified peptides, two novel sequences with potent ACE inhibitory activity were found. The amino acid sequences of these inhibitors were identified as Arg-Ala-Asp-His-Pro-Phe-Leu and Tyr-Ala-Glu-Glu-Arg-Tyr-Pro-Ile-Leu and showed IC50 values of 6.2 and 4.7 microM, respectively.     

Prevalence of Escherichia coli O157 in cattle and swine in central Mexico

Escherichia coli O157:H7 is a foodborne pathogenic bacterium that can reside undetected in the gastrointestinal tract of cattle because colonization by this bacterium is asymptomatic. Recent research has indicated that swine can carry and transmit this pathogen as well. The development of more advanced and sensitive detection techniques has improved the limit of detection and increased sensitivity for this important pathogen. This study was undertaken to determine the prevalence of E. coli O157 in cattle and swine in Mexico with the more sensitive detection technique of immunomagnetic bead separation. Samples (n = 60 per farm) were taken from four cattle and four swine farms (n = 240 cattle samples, n = 240 swine samples) located throughout central Mexico in October 2001. The prevalence of E. coli O157 was found to be only 1.25% on cattle farms and 2.1% on swine farms. The prevalence in cattle in this study is lower than that reported in the United States and could be related to the lower reported prevalence of E. coli O157 in humans in Mexico. However, further research is needed to verify prevalence throughout other regions of Mexico, as well as prevalence during other seasons of the year.     

Cellular localization and changes in expression of prolactin receptor isoforms in sheep ovary throughout the estrous cycle

The actions of prolactin (PRL) on target cells depend on the type of prolactin receptor (PRLr) predominantly expressed, particularly whether the long PRLr isoform is expressed. The aims of this study were to determine the cellular localization and the changes in expression of long and short PRLr isoforms in sheep ovary throughout the estrous cycle. Long and short PRLrs were localized mostly in the same ovarian cells. Maximum signal intensity, particularly for long PRLrs, was found in stromal cells surrounding primordial and primary follicles, and, for both PRLrs, in granulosa cells of preantral follicles and in luteal cells. Moderate signal intensity for PRLrs was found in theca cells of preantral to ovulatory follicles, and in granulosa cells of antral follicles up to the gonadotropin-dependent stage. Decreasing immunoreactivity to PRLrs was found in granulosa cells of gonadotropin-dependent to ovulatory follicles. For long PRLrs in particular, no signal was found in mural granulosa cells of gonadotropin-dependent follicles; for both isoforms, no signal was found in most granulosa cells of ovulatory follicles. In primordial to gonadotropin-dependent follicles, cellular localization of PRLr was similar on days 0, 10 and 15 of the cycle. Oocytes consistently showed positive immunostaining for PRLrs. Comparative RT-PCR analysis of long and short PRLr expression showed that the short isoform is evenly expressed throughout the estrous cycle, whereas the expression of the long form increases at the time of estrus and decreases at mid-luteal phase and at the onset of the follicular phase. Expression of long PRLrs was greater than that of short PRLrs on day 0 of cycle; expression of both isoforms was similar on day 10 and on day 15, long PRLrs expression was lower than that of short PRLrs. Our results indicate that in sheep ovary, the maximum responsiveness to PRL might occur during the preovulatory phase of the estrous cycle.     

Antimicrobial and antioxidant activities of natural extracts in vitro and in ground beef

Inhibition of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes by grape seed extract (ActiVin) and pine bark extract (Pycnogenol) and the effect of these natural extracts on the oxidative stability of raw ground beef were studied. In an agar dilution test, the MICs of ActiVin and Pycnogenol were determined to be 4.0 mg/ml for 4.43 log CFU per plate of E. coli O157:H7 and 4.0 mg/ml for 4.38 log CFU per plate of L. monocytogenes. In an inhibition curve test, populations of E. coli O157:H7, Salmonella Typhimurium, and L. monocytogenes fell to below the detection limit (10 CFU/ml) after 16 h of incubation. The numbers of E. coli O157:H7, L. monocytogenes, and Salmonella Typhimurium declined by 1.08, 1.24, and 1.33 log CFU/g, respectively, in raw ground beef treated with 1% Pycnogenol after 9 days of refrigerated storage. ActiVin (1%) and oleoresin rosemary (1%) resulted in an approximately 1-log CFU/g reduction in the populations of all three pathogens after 9 days. The addition of 1% ActiVin and Pycnogenol contributed to the maintenance of an acidic pH of 5.80 and 5.58, respectively, in raw ground beef. Compared to the control, all treatments increased in L* (lightness), with the exception of ActiVin. ActiVin and oleoresin rosemary had the highest a* (redness) and b* (yellowness) values, respectively. ActiVin most effectively retarded lipid oxidation, followed by Pycnogenol. The results suggest that these natural extracts have potential to be used with other preservative methods to reduce pathogenic numbers, lipid oxidation, and color degradation in ground beef.     

Differences in heat tolerance between preimplantation embryos from Brahman, Romosinuano, and Angus breeds

Exposure to 41 degrees C reduces development of embryos of heat-sensitive breeds (Holstein and Angus) more than for embryos of the heat-tolerant Brahman breed. Here it was tested whether embryonic resistance to heat shock occurs for a thermotolerant breed of different genetic origin than the Brahman. In particular, the thermal sensitivity of in vitro produced embryos of the Romosinuano, a Bos taurus, Criollo-derived breed, was compared to that for in vitro produced Brahman and Angus embryos. At d 4 after insemination, embryos > or = 8 cells were randomly assigned to control (38.5 degrees C) or heat shock (41 degrees C for 6 h) treatments. Heat shock reduced the proportion of embryos that developed to the blastocyst stage on d 8 after insemination. At 38.5 degrees C, there were no significant differences in development between breeds. Among embryos exposed to 41 degrees C, however, development was lower for Angus embryos than for Brahman and Romosinuano embryos. Furthermore, an Angus vs. (Brahman + Romosinuano) x temperature interaction occurred because heat shock reduced development more in Angus (30.3 +/- 4.6% at 38.5 degrees C vs. 4.9 +/- 4.6% at 41 degrees C) than in Brahman (25.1 +/- 4.6% vs. 13.6 +/- 4.6%) and Romosinuano (28.3 +/- 4.1% vs. 17.5 +/- 4.1%). Results demonstrate that embryos from Brahman and Romosinuano breeds are more resistant to elevated temperature than embryos from Angus. Thus, the process of adaptation of Brahman and Romosinuano breeds to hot environments resulted in both cases in selection of genes controlling thermotolerance at the cellular level.     

Ady2p is essential for the acetate permease activity in the yeast Saccharomyces cerevisiae

To identify new genes involved in acetate uptake in Saccharomyces cerevisiae, an analysis of the gene expression profiles of cells shifted from glucose to acetic acid was performed. The gene expression reprogramming of yeast adapting to a poor non-fermentable carbon source was observed, including dramatic metabolic changes, global activation of translation machinery, mitochondria biogenesis and the induction of known or putative transporters. Among them, the gene ADY2/YCR010c was identified as a new key element for acetate transport, being homologous to the Yarrowia lipolytica GPR1 gene, which has a role in acetic acid sensitivity. Disruption of ADY2 in S. cerevisiae abolished the active transport of acetate. Microarray analyses of ady2Delta strains showed that this gene is not a critical regulator of acetate response and that its role is directly connected to acetate transport. Ady2p is predicted to be a membrane protein and is a valuable acetate transporter candidate.