Obtention and characterization of phenolic extracts from different cocoa sources

The aim of this study was to evaluate several cocoa sources to obtain a rich phenol extract for use as an ingredient in the food industry. Two types of phenolic extracts, complete and purified, from different cocoa sources (beans, nibs, liquor, and cocoa powder) were investigated. UPLC-MS/MS was used to identify and quantify the phenolic composition of the extracts, and the Folin-Ciocalteu and vanillin assays were used to determine the total phenolic and flavan-3-ol contents, respectively. The DPPH and ORAC assays were used to measure their antioxidant activity. The results of the analysis of the composition of the extracts revealed that the major fraction was procyanidins, followed by flavones and phenolic acids. From the obtained results, the nib could be considered the most interesting source for obtaining a rich phenolic cocoa extract because of its rich phenolic profile content and high antioxidant activity in comparison with the other cocoa sources.     

A new method for selecting cytochimeras by the maximum number of nucleoli per cell in Allium wakegi Araki and A. fistulosum L.

Summary A new method for determining ploidy levels in cytochimeral plants was developed by examining the maximum number of nucleoli per cell. Colchicine-treated plants of Allium wakegi Araki and A. fistulosum L., which have different ploidy levels in the first (LI), the second (LII), and the third (LIII) germ layer, were used together with untreated 2-2-2 plants of the same species. Nucleoli in guard cells for LI and in mesophyll cells for LII were stained in a 50% AgNO₃ aqueous solution at 55° C for three hours under dark, humid conditions. In both species the ploidy levels of LI determined by the maximum number of nucleoli per guard cell accorded well with those determined by guard cell length. In A. fistulosum the ploidy levels of LII determined by the maximum number of nucleoli per mesophyll cell clearly agreed with those determined by pollen size. This method provided more precise and efficient identification for LI and LII ploidy than the conventional methods of using guard cell length for LI and pollen size for LII.     

Coral communities of Biscayne Bay,Florida and adjacent offshore areas: diversity,abundance, distribution,and environmental correlates

• 1. Hardbottom habitats of Biscayne Bay, a shallow lagoon adjacent to the city of Miami, Florida, USA, contain a limited number of coral species that represent a small subset of the species found at nearby offshore hardbottom and reef habitats of the Florida Reef Tract. Although the physical characteristics of this basin make it a marginal environment for coral growth, the presence of dense populations of Siderastrea radians and Porites furcata indicate that these, as well as other corals that are found at lower densities, are able to tolerate extreme and fluctuating conditions. Three factors, temperature, sedimentation, and salinity, appear to limit coral abundance, diversity, and distribution within Biscayne Bay.
• 2. Temperatures exhibit high frequencies of extreme high and low values known to cause coral stress and mortality elsewhere. Similarly, sedimentation rates are very high and sediment resuspension caused by currents, storms and boating activities commonly bury corals under sediment layers. Sediment burial was shown experimentally to influence growth and mortality of S. radians.
• 3. The salinity of Biscayne Bay is influenced by freshwater inputs from canal, sheetflow and groundwater sources that create a near‐shore environment with low mean salinity and high salinity fluctuation. Coral communities along this western margin have the lowest coral density and species richness. Chronic exposure to low salinity was shown experimentally to cause a decrease in the growth of S. radians.
• 4. The location of Biscayne Bay, downstream of a large restoration effort planned for the Everglades watershed, highlights the need to understand the relationship between the physical environment and the health of benthic communities. The data presented here provide the type of scientific information needed so that management decisions can take into account the potential impacts of human activities on the health of coral populations that are already near their tolerance limits for temperature, salinity, and sedimentation.

Copyright © 2003 John Wiley & Sons, Ltd.     

Detection and differentiation of several food-spoilage lactic acid bacteria by multiplex polymerase chain reaction, capillary gel electrophoresis, and laser-induced fluorescence

In this work, a complete analytical procedure is investigated to differentiate several food-spoilage lactic acid bacteria. To do that, a method involving multiplex Polymerase Chain Reaction (PCR), capillary gel electrophoresis (CGE), and laser-induced fluorescence (LIF) is developed. The PCR-CGE-LIF protocol allows the simultaneous detection and differentiation of the genera Leuconostoc and Carnobacterium, the nonmotile group of species within the genus Carnobacterium, and the three species of the group individually (C. divergens, C. gallinarum, and C. maltaromicum). The capability of this approach is clearly illustrated through the sensitive and efficient analysis of the two closest amplicons, with sizes equal to 397 and 412 bp, showing very different yields in all of the amplification reactions tested. These two fragments, which could not be resolved by agarose gel electrophoresis (AGE), are clearly distinguishable by CGE-LIF even when very different areas for both peaks are obtained. The PCR-CGE-LIF method also allows the sensitive detection of these bacteria, demonstrating both a significant resolution improvement compared with traditional AGE and the usefulness of this approach to solve real-life analytical challenges. Good reproducibility of the CGE-LIF procedure is shown for the analysis of multiplex PCR samples with percent relative standard deviation values for migration times and corrected peak areas as low as 0.80 and 6.50 for the same sample and three different days (n = 12), respectively.     

Interactions between bacteria and Cryptosporidium molnari in gilthead sea bream (Sparus aurata) under farm and laboratory conditions

The possible interaction of Cryptosporidium molnari and bacteria in gilthead sea bream (Sparus aurata) was studied. Epidemiological data from a pathological survey under farm conditions were analyzed. In addition, parasite and bacteria burdens were studied in experimental models in which naturally and experimentally parasitized fish were challenged with a particular strain of Vibrio harveyi (H57). All the bacteria species present were studied. Under farm conditions, the parasite was more prevalent when mortality or morbidity cases (study C) occurred than in randomly sampled fish (study B). In study C, parasite abundance was significantly higher in bacteria-negative fish, and total bacteria abundance was significantly higher within non-parasitized fish. V. harveyi and V. splendidus were the most prevalent among bacteria carriers in studies B and C, respectively. In study C, among bacteria carriers, most isolates were slightly more prevalent in parasitized than in non-parasitized fish. Two groups (G1, G2) of naturally parasitized fish were inoculated with H57 by intracoelomic injection (ICI) and by oral intubation (OI). H57 was recovered only in G1 inoculated fish, which had a significantly higher basal abundance of total bacteria, and where the only ones with mortalities. In G1, the mortality rate and the prevalence of other V. harveyi strains different from the H57 molecular type were higher in ICI than in OI fish, and the total bacteria abundance was also significantly higher in ICI fish. C. molnari abundance was significantly higher in G1 than in G2, and also in OI than in ICI fish within G1. When H57 was IC inoculated to fish (G3, from the same farm as G2) experimentally infected with C. molnari, H57 was not recovered from any fish. A low mortality was recorded, and only in those fish inoculated with both pathogens. Also in these fish, the prevalence of infection of C. molnari was higher and histopathological damage to the stomach was greater than in fish inoculated only with the parasite. Therefore, the impact of the parasite would be reduced notably when the bacterial burden or the intensity of parasite infection are low (G2, G3).     

Nitrogen turnover in bare soil planted subsequently with grass as investigated by electro‐ultrafiltration (EUF)

Changes of EUF‐extractable nitrogen (N) (nitrate, ammonium, organic N) in 20 arable bare soils, subsequently planted with ryegrass (Lolium multiflorum L.) and cutting three times were investigated in pot experiments. All 20 soils responded qualitatively in the same way. During the period of bare soil, there was a significant increase of EUF‐extractable nitrate (EUF NO ), while extractable ammonium (EUF NH ) remained on the same level and organic N (EUF N_org) decreased. This decrease, however, was not significant. From sowing until the first cutting of the grass, EUF‐NO concentration decreased to almost zero. This low EUF‐NO level was maintained throughout the subsequent experimental period (three cuttings of grass). During the growth of the first cutting, EUF N_org decreased while EUF NH remained constant, however, on a low level. EUF NH fell during the growth of the second and third cutting. In this period, however, the N supply of the grass was insufficient. EUF N_org decreased during the growth of the second cutting, but increased during the growth of the third cutting. This shows that the EUF‐N_org fraction represents a transient pool, which gains and loses N. EUF NO , EUF NH , and EUF N_org correlated with the N uptake of the grass. Strongest correlation for EUF NO was found for the first cutting (p < 0.001), and for EUF NH and EUF N_org for the second and third cutting (p < 0.001). Total soil N was not correlated with the N uptake of the grass. EUF N_org was only about 2% of the total N. This relatively small EUF‐N_org fraction, however, is relevant for the mineralization of organic soil N, and the N quantity indicated by EUF N_org is in the range of the N amount mineralized in arable soils within a growing season.     

Development of a phenol-enriched olive oil with both its own phenolic compounds and complementary phenols from thyme

Besides affecting the oil's sensorial characteristics, the presence of herbs and spices has an impact on the nutritional value of the flavored oils. The aim of the study was to develop a new product based on the phenol-enrichment of a virgin olive oil with both its own phenolic compounds (secoiridoid derivatives) plus additional complementary phenols from thyme (flavonoids). We studied the effect of the addition of phenolic extracts (olive cake and thyme) on phenolic composition, oxidative stability, antioxidant activity, and bitter sensory attribute of olive oils. Results showed that flavonoids from thyme appeared to have higher transference ratios (average 89.7%) from the phenolic extract to oil, whereas secoiridoids from olive presented lower transference ratios (average 35.3%). The bitter sensory attribute of the phenol-enriched oils diminished with an increase of the concentration of phenols from thyme, which might denote an improvement in the consumer acceptance.