In-depth mechanistic study on the formation of acrylamide and other vinylogous compounds by the maillard reaction

The formation of acrylamide was studied in low-moisture Maillard model systems (180 degrees C, 5 min) based on asparagine, reducing sugars, Maillard intermediates, and sugar degradation products. We show evidence that certain glycoconjugates play a major role in acrylamide formation. The N-glycosyl of asparagine generated about 2.4 mmol/mol acrylamide, compared to 0.1-0.2 mmol/mol obtained with alpha-dicarbonyls and the Amadori compound of asparagine. 3-Hydroxypropanamide, the Strecker alcohol of asparagine, generated only low amounts of acrylamide ( approximately 0.23 mmol/mol), while hydroxyacetone increased the acrylamide yields to more than 4 mmol/mol, indicating that alpha-hydroxy carbonyls are much more efficient than alpha-dicarbonyls in converting asparagine into acrylamide. The experimental results are consistent with the reaction mechanism based on (i) a Strecker type degradation of the Schiff base leading to azomethine ylides, followed by (ii) a beta-elimination reaction of the decarboxylated Amadori compound to afford acrylamide. The beta-position on both sides of the nitrogen atom is crucial. Rearrangement of the azomethine ylide to the decarboxylated Amadori compound is the key step, which is favored if the carbonyl moiety contains a hydroxyl group in beta-position to the nitrogen atom. The beta-elimination step in the amino acid moiety was demonstrated by reacting under low moisture conditions decarboxylated model Amadori compounds obtained by synthesis. The corresponding vinylogous compounds were only generated if a beta-proton was available, for example, styrene from the decarboxylated Amadori compound of phenylalanine. Therefore, it is suggested that this thermal pathway may be common to other amino acids, resulting under certain conditions in their respective vinylogous reaction products.     

Reduction processes in forest wetlands: Tracking down heterogeneity of source/sink functions with a combination of methods

Wetlands are considered to be the biggest unknowns regarding the influence of global climate change on element dynamics, so knowledge of processes and conditions controlling sink and source functions of redox processes is crucial. The aim of this study was to investigate the sink/source function of nitrate, Fe, sulfate reduction and methanogenesis of an upland and a lowland fen within a boreal spruce catchment, southern Germany. We used suction cups and anaerobic dialysis chambers for soil solution sampling, FeS probes for the determination of S oxidation potential and stability of anoxic conditions and analysis of the soil solid phase (contents of C, S and Fe species). Both fens had high rates of nitrate reduction and potentially high rates of CH₄ production. The upper few cm of all profiles were oxic with low CH₄ concentrations, suggesting low CH₄ emission rates from the soil, though emission by vascular plants cannot be excluded. Sulfate and Fe reduction processes differed significantly in the fens. The upland fen was characterized by relatively stable anoxic conditions, low Fe contents but high contents of organic S and low C/S ratios. We concluded that the upland fen is an effective sink for sulfate with long-term S storage. In contrast, the lowland fen was characterized by alternating oxidation-reduction cycles with high Fe contents, lower contents of organic S and higher C/S ratios. Thus, even though low sulfate and high Fe concentrations in soil solutions indicated high reduction rates in the lowland fen, long-term storage of S is not likely in this fen. Differences in biogeochemical processes between sites are most likely not associated with hydrology but rather with the role of vascular plants.     

Quantitation of ochratoxin A in South African wines

The natural occurrence of the carcinogenic mycotoxin ochratoxin A (OTA) in wines sold in local retail outlets in South Africa and Italy was investigated by HPLC analysis with fluorescence detection following cleanup by immunoaffinity column. All 24 local South African wines tested (15 white and 9 red) were found to contain detectable levels (>0.01 microg/L) of OTA, with a mean of 0.16 microg/L in the white wines and a mean of 0.24 microg/L in the red wines. Results were subsequently confirmed by LC-MS analysis using positive ion electrospray ionization with collision-induced dissociation of the protonated molecular ion [M + H](+) at m/z 404 and selected reaction monitoring of the resultant product ions [M + H - H(2)O - CO](+) at m/z 358 and [M + H - H(2)O](+) at m/z 386. Comparison with the fluorescence method gave a significant correlation (r = 0.87; p < 0.01). Although OTA contamination was present in all of the South African samples analyzed, levels were well below the suggested European Union limit of 0.5 microg/kg. The highest level found in a locally purchased wine was 0.39 microg/L in a blend of local and imported Spanish red wine. Of the eight Italian wines analyzed, only two red wines were contaminated above the suggested maximum level.     

Associations between immune competence phenotype and feedlot health and productivity in Angus cattle

Genetic strategies aimed at improving general immune competence (IC) have the potential to reduce the incidence and severity of disease in beef production systems, with resulting benefits of improved animal health and welfare and reduced reliance on antibiotics to prevent and treat disease. Implementation of such strategies first requires that methodologies be developed to phenotype animals for IC and demonstration that these phenotypes are associated with health outcomes. We have developed a methodology to identify IC phenotypes in beef steers during the yard weaning period, which is both practical to apply on-farm and does not restrict the future sale of tested animals. In the current study, a total of 838 Angus steers, previously IC phenotyped at weaning, were categorized as low (n = 98), average (n = 653), or high (n = 88) for the IC phenotype. Detailed health and productivity data were collected on all steers during feedlot finishing, and associations between IC phenotype, health outcomes, and productivity were investigated. A favorable association between IC phenotype and number of mortalities during feedlot finishing was observed with higher mortalities recorded in low IC steers (6.1%) as compared with average (1.2%, P < 0.001) or high (0%, P = 0.018) IC steers. Disease incidence was numerically highest in low IC steers (15.3 cases/100 animals) and similar in average IC steers (10.1 cases/100 animals) and high IC steers (10.2 cases/100 animals); however, differences between groups were not significant. No significant influence of IC phenotype on average daily gain was observed, suggesting that selection for improved IC is unlikely to incur a significant penalty to production. The potential economic benefits of selecting for IC in the feedlot production environment were calculated. Health-associated costs were calculated as the sum of lost production costs, lost capital investment costs, and disease treatment costs. Based on these calculations, health-associated costs were estimated at AUS$103/head in low IC steers, AUS$25/head in average IC steers, and AUS$4/head in high IC steers, respectively. These findings suggest that selection for IC has the potential to reduce mortalities during feedlot finishing and, as a consequence, improve the health and welfare of cattle in the feedlot production environment and reduce health-associated costs incurred by feedlot operators.     

Lipid composition of Collembola and their food resources in deciduous forest stands—Implications for feeding strategies

The lipid composition of Collembola and their potential food resources was assessed in three deciduous forest stands, in order to gain insight into food web linkages under field conditions. Fatty acids (FAs) previously assigned as trophic markers in laboratory experiments were used to investigate feeding strategies in situ. As potential food sources soil microbiota and plant debris were characterised by their phospholipid fatty acid (PLFA) composition. Both the amount and the pattern of PLFAs differed between sites and soil depth, in particular the bacterial and plant marker FAs in the upper soil layer. Thus, the availability of resources for micro-detritivores varied due to forest and soil layer. The lipid composition of vital and senescent beech leaves was predominantly influenced by metabolic status and represented a quite homogenous FA resource across forest stands. Comparing Folsomia quadrioculata, Lepidocyrtus lignorum, Neanura muscorum and Pogonognathellus longicornis between the different forests revealed FA profiles to be predominantly affected by site, suggesting a diet shift depending on resources at hand. However, species-specific differences in individual FAs occurred, likely related to feeding strategy and physiological activity. Lipids of Collembola comprised low amounts of bacterial marker FAs, and bacterial consumption may occur to some extent, particularly on Gram-positives. The marker FA for predatory feeding, 20:1ω9, was found in several species, although in low amounts. This contradicts known feeding habits and caution is advisable in using 20:1ω9 as trophic marker. Overall, as indicated by high proportions of oleic (18:1ω9) or linoleic (18:2ω6,9) acid, most species were either plant litter or fungal feeders, with some transitions. The ratio 18:1ω9/18:2ω6,9 is proposed as a tool to distinguish between these two major feeding strategies in Collembola.     

BPV-1 infection is not confined to the dermis but also involves the epidermis of equine sarcoids

In equids, bovine papillomaviruses of type 1 (BPV-1) and less frequently type 2 induce common, locally aggressive skin tumours termed sarcoids. Whereas BPV infection in cattle usually involves the epidermis and is productive in this skin layer, infection in equids is currently thought to be abortive, with virus solely residing as multiple episomes in dermal fibroblasts. Based on recent observations that do not agree with this assumption, we hypothesised that BPV also infects equid epidermis and is active in this skin layer. To test this hypothesis, we conducted a proof-of-principle study on eight distinct sarcoids. Presence of viral DNA was addressed by qualitative and quantitative BPV-1 PCR from microdissected sarcoid epidermis, and by subsequent amplicon sequencing. Viral activity was assessed by screening sarcoid epidermis for BPV-1 protein expression using immunohistochemistry (IHC) or immunofluorescence (IF). Virus-free equine skin served as negative control throughout the assays. BPV-1 DNA was demonstrated in all sarcoid epidermis samples, with viral DNA loads ranging between 2 and 195 copies/cell. Identical BPV-1 E5 genes were identified in epidermis and dermis of each of two sarcoids, yet different E5 variants were found in individual lesions. IHC/IF revealed the presence of E5 and E7 protein in sarcoid epidermis, and L1 capsomers in the squamous layer of one lesion. These findings indicate that BPV infection also involves the epidermis, where it may occasionally be productive.