Preparation and comparative release characteristics of three anthocyanin encapsulation systems

Bilberries (Vaccinium myrtillus L.) and their major polyphenolic constituents, anthocyanins, have preventive activities inter alia against colon cancer and inflammatory bowel diseases. However, anthocyanins are sensitive to environmental conditions; thus their bioavailability in the gastrointestinal tract is an important determinant of their in vivo activity. In the study reported here, the potential benefits of encapsulating an anthocyanin rich bilberry extract (BE) on anthocyanin stability were investigated. Nonencapsulated BE and three different BE loaded microcapsule systems were incubated in simulated gastric fluid (SGF) and fed state simulated intestinal fluid (FeSSIF). After exposure to these media, released anthocyanins were identified and quantified by HPLC with UV/Vis detection. Although a rapid release of anthocyanins was observed within the first 20 min, encapsulation of anthocyanins doubled the amount of available anthocyanins after 150 min of incubation. These results illustrate the ability of encapsulation to inhibit early degradation of anthocyanins in the intestinal system.     

Soil‐ecological evaluation of willows in a floodplain

Willows (Salix spp.) were supposed to be suitable candidates for the phytoremediation of polluted floodplain soils, but it is unknown how willow growth alters concentrations and mobility of pollutants under the conditions of ongoing periodically flooding. Therefore, effects of willow cropping on total concentrations and mobility of As and heavy metals and soil microbial properties were determined after three and four growing seasons under willows in comparison to native grassland in a flood channel of a River Elbe floodplain (Central Germany). After 4 y of willow coppice, the heavy metal concentrations (mg kg^–1) were increased not only in the grass control plots (final Cu 274, Pb 276, Zn 935) but also under the willows (final Cu 248, Pb 251, Zn 779) compared to the initial concentrations (initial Cu 170, Pb 156, Zn 579). This increase might likely be caused by the ongoing sedimentation by flood events. The smaller increase under willows compared to grass might be related to an initial net effect of phytoextraction. The concentrations of the mobile fractions of Cd, Cu, Ni, and Zn were significantly lower under willows than under grass. Higher β‐glucosidase activities under willows than under grass might indicate a starting net decomposition of organic matter. Therefore, the study of long‐term and large‐scale effects are recommend before an appropriate evaluation of willow short‐rotation coppice for phytoremediation of polluted floodplains will be established.     

Examining the knowledge base and status of commercially exploited marine species with the RAM Legacy Stock Assessment Database

Meta‐analyses of stock assessments can provide novel insight into marine population dynamics and the status of fished species, but the world’s main stock assessment database (the Myers Stock‐Recruitment Database) is now outdated. To facilitate new analyses, we developed a new database, the RAM Legacy Stock Assessment Database, for commercially exploited marine fishes and invertebrates. Time series of total biomass, spawner biomass, recruits, fishing mortality and catch/landings form the core of the database. Assessments were assembled from 21 national and international management agencies for a total of 331 stocks (295 fish stocks representing 46 families and 36 invertebrate stocks representing 12 families), including nine of the world’s 10 largest fisheries. Stock assessments were available from 27 large marine ecosystems, the Caspian Sea and four High Seas regions, and include the Atlantic, Pacific, Indian, Arctic and Antarctic Oceans. Most assessments came from the USA, Europe, Canada, New Zealand and Australia. Assessed marine stocks represent a small proportion of harvested fish taxa (16%), and an even smaller proportion of marine fish biodiversity (1%), but provide high‐quality data for intensively studied stocks. The database provides new insight into the status of exploited populations: 58% of stocks with reference points (n = 214) were estimated to be below the biomass resulting in maximum sustainable yield (B_MSY) and 30% had exploitation levels above the exploitation rate resulting in maximum sustainable yield (U_MSY). We anticipate that the database will facilitate new research in population dynamics and fishery management, and we encourage further data contributions from stock assessment scientists.     

Chronic pneumonia in calves after experimental infection with Mycoplasma bovis strain 1067: Characterization of lung pathology,persistence of variable surface protein antigens and local immune response

Background

Mycoplasma bovis is associated with pneumonia in calves characterized by the development of chronic caseonecrotic lesions with the agent persisting within the lesion. The purposes of this study were to characterize the morphology of lung lesions, examine the presence of M. bovis variable surface protein (Vsp) antigens and study the local immune responses in calves after infection with M. bovis strain 1067.

Methods

Lung tissue samples from eight calves euthanased three weeks after experimental infection with M. bovis were examined by bacteriology and pathology. Lung lesions were evaluated by immunohistochemical (IHC) staining for wide spectrum cytokeratin and for M. bovis Vsp antigens and pMB67 antigen. IHC identification and quantitative evaluation of CD4⁺ and CD8⁺ T lymphocytes and immunoglobulin (IgG1, IgG2, IgM, IgA)-containing plasma cells was performed. Additionally, expression of major histocompatibility complex class II (MHC class II) was studied by IHC.

Results

Suppurative pneumonic lesions were found in all calves. In two calves with caseonecrotic pneumonia, necrotic foci were surrounded by epithelial cells resembling bronchial or bronchiolar epithelium. In all calves, M. bovis Vsp antigens were constantly present in the cytoplasm of macrophages and were also present extracellularly at the periphery of necrotic foci. There was a considerable increase in numbers of IgG1- and IgG2-positive plasma cells among which IgG1-containing plasma cells clearly predominated. Statistical evaluation of the numbers of CD4⁺ and CD8⁺ T cells, however, did not reveal statistically significant differences between inoculated and control calves. In M. bovis infected calves, hyperplasia of bronchus-associated lymphoid tissue (BALT) was characterized by strong MHC class II expression of lymphoid cells, but only few of the macrophages demarcating the caseonecrotic foci were positive for MHC class II.

Conclusions

The results from this study show that infection of calves with M. bovis results in various lung lesions including caseonecrotic pneumonia originating from bronchioli and bronchi. There is long-term persistence of M. bovis as demonstrated by bacteriology and immunohistochemistry for M. bovis antigens, i.e. Vsp antigens and pMB67. The persistence of the pathogen and its ability to evade the specific immune response may in part result from local downregulation of antigen presenting mechanisms and an ineffective humoral immune response with prevalence of IgG1 antibodies that, compared to IgG2 antibodies, are poor opsonins.     

Mapping adaptation of barley to droughted environments

Identifying barley genomic regions influencing the response of yield and its components to water deficits will aid in our understanding of the genetics of drought tolerance and the development of more drought tolerant cultivars. We assembled a population of 192 genotypes that represented landraces, old, and contemporary cultivars sampling key regions around the Mediterranean basin and the rest of Europe. The population was genotyped with a stratified set of 50 genomic and EST derived molecular markers, 49 of which were Simple Sequence Repeats (SSRs), which revealed an underlying population sub-structure that corresponded closely to the geographic regions in which the genotypes were grown. A more dense whole genome scan was generated by using Diversity Array Technology (DArT®) to generate 1130 biallelic markers for the population. The population was grown at two contrasting sites in each of seven Mediterranean countries for harvest 2004 and 2005 and grain yield data collected. Mean yield levels ranged from 0.3 to 6.2 t/ha, with highly significant genetic variation in low-yielding environments. Associations of yield with barley genomic regions were then detected by combining the DArT marker data with the yield data in mixed model analyses for the individual trials, followed by multiple regression of yield on markers to identify a multi-locus subset of significant markers/QTLs. QTLs exhibiting a pre-defined consistency across environments were detected in bins 4, 6, 6 and 7 on barley chromosomes 3H, 4H, 5H and 7H respectively.     

QTLs for chlorophyll and chlorophyll fluorescence parameters in barley under post-flowering drought

Drought is one of the major factors limiting barley yields in many developing countries worldwide. The identification of molecular markers linked to genes controlling drought tolerance in barley is one way to improve breeding efficiency. In this study, we analyzed the quantitative trait loci (QTL) controlling chlorophyll content and chlorophyll fluorescence in 194 recombinant inbred lines (RILs) developed from the cross between the cultivar ‘Arta’ and Hordeum spontaneum 41-1. Five traits, chlorophyll content, and four chlorophyll fluorescence parameters, namely initial fluorescence (F_o), maximum fluorescence (F_m), variable fluorescence (F_v), and maximum quantum efficiency of PSII (F_v/F_m) which are related to the activity of the photosynthetic apparatus, were measured under well-watered and drought stress conditions at post-flowering stage. QTL analysis identified a total of nine and five genomic regions, under well-watered and drought stress conditions, respectively, that were significantly associated with the expression of the five target traits at post-flowering stage. No common QTL was detected except one for chlorophyll content, which was identified in both growth conditions, demonstrating that the genetic control of the expression of the traits related to photosynthesis differed under different water conditions. A QTL for F_v/F_m, which is related to the drought tolerance of photosynthesis was identified on chromosome 2H at 116 cM in the linkage map under drought stress. This QTL alone explained more than 15% of phenotypic variance of maximum quantum yield of PSII, and was also associated with the expression of four other traits. In addition, another QTL for F_v/F_m was also located on the same chromosome (2H) but at 135.7 cM explaining around 9% of the phenotypic variance under drought conditions. The result presented here suggest that two major loci, located on chromosome 2H, are involved in the development of functional chloroplast at post-flowering stage for drought tolerance of photosynthesis in barley under drought stress. If validated in other populations, chlorophyll fluorescence parameters could be used as selection criteria for drought tolerance.     

DON content in oat grains in Norway related to weather conditions at different growth stages

High concentrations of the mycotoxin deoxynivalenol (DON), produced by Fusarium graminearum have occurred frequently in Norwegian oats recently. Early prediction of DON levels is important for farmers, authorities and the Cereal Industry. In this study, the main weather factors influencing mycotoxin accumulation were identified and two models to predict the risk of DON in oat grains in Norway were developed: (1) as a warning system for farmers to decide if and when to treat with fungicide, and (2) for authorities and industry to use at harvest to identify potential food safety problems. Oat grain samples from farmers’ fields were collected together with weather data (2004–2013). A mathematical model was developed and used to estimate phenology windows of growth stages in oats (tillering, flowering etc.). Weather summarisations were then calculated within these windows, and the Spearman rank correlation factor calculated between DON-contamination in oats at harvest and the weather summarisations for each phenological window. DON contamination was most clearly associated with the weather conditions around flowering and close to harvest. Warm, rainy and humid weather during and around flowering increased the risk of DON accumulation in oats, as did dry periods during germination/seedling growth and tillering. Prior to harvest, warm and humid weather conditions followed by cool and dry conditions were associated with a decreased risk of DON accumulation. A prediction model, including only pre-flowering weather conditions, adequately forecasted risk of DON contamination in oat, and can aid in decisions about fungicide treatments.     

Production of Chitooligosaccharides and Their Potential Applications in Medicine

Chitooligosaccharides (CHOS) are homo- or heterooligomers of N-acetylglucosamine and D-glucosamine. CHOS can be produced using chitin or chitosan as a starting material, using enzymatic conversions, chemical methods or combinations thereof. Production of well-defined CHOS-mixtures, or even pure CHOS, is of great interest since these oligosaccharides are thought to have several interesting bioactivities. Understanding the mechanisms underlying these bioactivities is of major importance. However, so far in-depth knowledge on the mode-of-action of CHOS is scarce, one major reason being that most published studies are done with badly characterized heterogeneous mixtures of CHOS. Production of CHOS that are well-defined in terms of length, degree of N-acetylation, and sequence is not straightforward. Here we provide an overview of techniques that may be used to produce and characterize reasonably well-defined CHOS fractions. We also present possible medical applications of CHOS, including tumor growth inhibition and inhibition of T_H2-induced inflammation in asthma, as well as use as a bone-strengthener in osteoporosis, a vector for gene delivery, an antibacterial agent, an antifungal agent, an anti-malaria agent, or a hemostatic agent in wound-dressings. By using well-defined CHOS-mixtures it will become possible to obtain a better understanding of the mechanisms underlying these bioactivities.     

Mycorrhizal community structure,microbial biomass P and phosphatase activities under Salix polaris as influenced by nutrient availability

Low supply of the nutrients nitrogen (N) and phosphorus (P) limit plant growth and spreading, and increase the plant-microbial nutrient competition in subarctic and arctic regions. We investigated the mycorrhizal community structure of a polar shrub willow (Salix polaris) and the microbial turnover in its rhizosphere to explore the adaptation of a mycorrhizal plant in the subarctic tundra. The ectomycorrhizal colonisation ranged from 35 to 64% of the fine root tips and decreased with an increasing soil C/N ratio. In total, 16 ectomycorrhizal morphotypes were found under S. polaris (eight to 13 morphotypes per site, five morphotypes at all four sites). Cenococcum sp. was the most common EM fungus (32% of the ectomycorrhizal fine roots). The abundance of Cenococcum sp. increased with an increasing organic matter content and N/P ratio in the soil. Arbuscular mycorrhizal colonisation of S. polaris was absent or less than 1% of the fine root length. Microbial biomass P accounted for 21–75% of the organic soil P and 6–49% of the total soil P. Microbial biomass P, alkaline and acid phosphatase activities in the rhizosphere increased with increasing soil N concentration. We conclude that a higher N supply decreases the diversity in the mycorrhizal community on polar willows and increases the role of P turnover from the soil microbial biomass for the nutrient supply.