Uncertainties in the determination of pyrrolizidine alkaloid levels in naturally contaminated honeys and comparison of results obtained by different analytical approaches

The contamination of honey with hepatotoxic pyrrolizidine alkaloids (PAs) is a well-known hazard for food safety. While management strategies and controls of the honey industry aim to reduce the PA levels, uncertainties remain with regard to the safety of regionally produced and marketed honey. In addition, a previous study showed large differences of results obtained after various periods of storage and apparent differences between the analytical results of different laboratories. Therefore, this study aimed at examining these uncertainties by monitoring the impact of storage on the PA and PA N-oxide (PANO) content of two freshly harvested honeys and on possible demixing effects caused by pollen settling. Additionally, three analytical approaches – target analysis with matrix-matched calibration or standard addition and a sum parameter method – were applied for a comparative analysis of 20 honeys harvested in summer 2016. All samples originated from Schleswig-Holstein in Northern Germany where the PA plant Jacobaea vulgaris is currently observed on a massive scale. The results of the time series analyses showed that PANO levels markedly decreased within a few weeks and practically reached the LOD 16 weeks after harvest. Tertiary PAs, by contrast, remained stable and did not increase as a consequence of PANO decrease. The experiments on a putative demixing, which may result in a heterogeneous distribution of PAs/PANOs, revealed that there was no such effect during storage of up to 12 weeks. A comparison of the PA/PANO levels obtained by different analytical approaches showed that in some cases the sum parameter method yielded much higher levels than the target approaches, whereas in other cases, the target analysis with standard addition found higher levels than the other two methods. In summary, the results of this study highlight uncertainties regarding the validity and comparability of analytical results and consequently regarding health risk assessment.     

Automated DNA preparation from maize tissues and food samples suitable for real-time PCR detection of native genes

There is an increasing need for high-throughput analyses of plants and food samples for the presence of specific DNA sequences, e.g. transgenic contaminations. We developed and optimized conditions for the automated isolation of DNA from several maize tissues and various edibles containing maize using the MagNA Pure LC system (Roche Applied Science). Our results show that the system provided is capable of isolating DNA from any tested source. Quantification of an endogenous gene by LightCycler real-time PCR revealed that the DNA is suitable in quality and quantity for multiple PCR analyses.     

Hypoxia in the Baltic Sea and basin-scale changes in phosphorus biogeochemistry

Deep-water oxygen concentrations in the Baltic Sea are influenced by eutrophication, but also by saltwater inflows from the North Sea. In the last two decades, only two major inflows have been recorded and the lack of major inflows is believed to have resulted in a long-term stagnation of the deepest bottom water. Analyzing data from 1970 to 2000 at the basin scale, we show that the estimated volume of water with oxygen, <2 mL L(-1), was actually at a minimum at the end of the longest so-called stagnation period on record. We also show that annual changes in dissolved inorganic phosphate water pools were positively correlated to the area of bottom covered by hypoxic water, but not to changes in total phosphorus load, thus addressing the legacy of eutrophication on a basinwide scale. The variations in phosphorus pools that have occurred during the past decades do not reflect any human action to reduce inputs. The long residence time and internally controlled variation of the large P pool in the Baltic Sea has important implications for management of both N and P inputs into this eutrophicated enclosed basin.     

Predicting adult fish acute lethality with the zebrafish embryo: relevance of test duration, endpoints, compound properties, and exposure concentration analysis

The zebrafish embryo toxicity test has been proposed as an alternative for the acute fish toxicity test, which is required by various regulations for environmental risk assessment of chemicals. We investigated the reliability of the embryo test by probing organic industrial chemicals with a wide range of physicochemical properties, toxicities, and modes of toxic action. Moreover, the relevance of using measured versus nominal (intended) exposure concentrations, inclusion of sublethal endpoints, and different exposure durations for the comparability with reported fish acute toxicity was explored. Our results confirm a very strong correlation of zebrafish embryo to fish acute toxicity. When toxicity values were calculated based on measured exposure concentrations, the slope of the type II regression line was 1 and nearly passed through the origin (1 to 1 correlation). Measured concentrations also explained several apparent outliers. Neither prolonged exposure (up to 120 h) nor consideration of sublethal effects led to a reduced number of outliers. Yet, two types of compounds were less lethal to embryos than to adult fish: a neurotoxic compound acting via sodium channels (permethrin) and a compound requiring metabolic activation (allyl alcohol).     

Characterizing biofuel combustion with patterns of real-time emission data (PaRTED)

Emission properties and quantities from combustion sources can vary significantly during operation, and this characteristic variability is hidden in the traditional presentation of emission test averages. As a complement to the emission test averages, we introduce the notion of statistical pattern analysis to characterize temporal fluctuations in emissions, using cluster analysis and frequency plots. We demonstrate this approach by comparing emissions from traditional and improved wood-burning cookstoves under in-field conditions, and also to contrast laboratory and in-field cookstove performance. Compared with traditional cookstoves, improved cookstoves eliminate emissions that occur at low combustion efficiency. For cookstoves where the only improvement is an insulated combustion chamber, this change results in emission of more light-absorbing (black) particles. When a chimney is added, the stoves produce more black particles but also have reduced emission factors. Laboratory tests give different results than in-field tests, because they fail to reproduce a significant fraction of low-efficiency events, spikes in particulate matter (PM) emissions, and less-absorbing particles. These conditions should be isolated and replicated in future laboratory testing protocols to ensure that stove designs are relevant to in-use operation.     

Telephone booster sessions for optimizing smoking cessation for patients in rehabilitation centers.

Smokers with smoking-related diseases who are hospitalized in rehabilitation centers should be offered smoking cessation. This is the first study evaluating whether telephone booster sessions after intensive inpatient treatment are an effective strategy. The present study was conducted in 13 rehabilitation centers for somatic disorders as a prospective multicenter study with a randomized treatment-control group design. We compared abstinence rates after hospital discharge from treatment that included a group smoking cessation program with (treatment group) and without telephone booster sessions (control group). Data from 290 smokers were analyzed. After 6 and 12 months the treatment group achieved abstinence rates twice as high as those of the control group. Men profited more from telephone booster sessions than did women. Results indicated that telephone booster sessions were highly effective (even) after an inherently intensive group program during a hospital stay. Further research should focus on the special needs of women receiving telephone counseling.     

Platinum and palladium emissions from on-road vehicles in the Kaisermühlen Tunnel (Vienna, Austria)

Total and size-segregated Pt and Pd emission factors from on-road vehicles were measured in the Kaisermühlen Tunnel in Vienna, Austria. Aerosol sampling was performed simultaneously inside and outside the tunnel during April and May 2005. Analysis of the acid-digested aerosol samples was performed using a preconcentration procedure with subsequent on-line detection by electro-thermal atomic absorption spectrometry (ETAAS). Inside the tunnel distinctly increased Pt and Pd concentrations were found with highest levels in total suspended particulate matter samples and reduced concentrations in the size-segregated PM10 and PM2.5 samples. Emission factors were calculated from concentration differences between tunnel inside and tunnel outside samples, the distance between tunnel entrance and sampling location, the ventilation rate, and the number of vehicles passing through the tunnel. Emission rates observed for Pt ranged from 38 +/- 5.9 to 146 +/- 13 ng veh(-1) km(-1), whereas the emission factors of Pd varied between 13 +/- 2.1 and 42 +/- 4.1 ng veh(-1) km(-1). Variations in the emission rates were assumed to originate from alterations in traffic conditions. Size-segregated investigations revealed that the major part of Pt and Pd emissions were released in the coarse aerosol mode (size fraction > PM10), nevertheless a considerable fraction (approximately 12% and approximately 22% respectively) was emitted in the inhalable PM2.5 fraction.     

Carbon and sulfur cycling by microbial communities in a gypsum-treated oil sands tailings pond

Oil sands tailings ponds receive and store the solid and liquid waste from bitumen extraction and are managed to promote solids densification and water recycling. The ponds are highly stratified due to increasing solids content as a function of depth but can be impacted by tailings addition and removal and by convection due to microbial gas production. We characterized the microbial communities in relation to microbial activities as a function of depth in an active tailings pond routinely treated with gypsum (CaSO(4)·2H(2)O) to accelerate densification. Pyrosequencing of 16S rDNA gene sequences indicated that the aerobic surface layer, where the highest level of sulfate (6 mM) but no sulfide was detected, had a very different community profile than the rest of the pond. Deeper anaerobic layers were dominated by syntrophs (Pelotomaculum, Syntrophus, and Smithella spp.), sulfate- and sulfur-reducing bacteria (SRB, Desulfocapsa and Desulfurivibrio spp.), acetate- and H(2)-using methanogens, and a variety of other anaerobes that have been implicated in hydrocarbon utilization or iron and sulfur cycling. The SRB were most abundant from 10 to 14 mbs, bracketing the zone where the sulfate reduction rate was highest. Similarly, the most abundant methanogens and syntrophs identified as a function of depth closely mirrored the fluctuating methanogenesis rates. Methanogenesis was inhibited in laboratory incubations by nearly 50% when sulfate was supplied at pond-level concentrations suggesting that in situ sulfate reduction can substantially minimize methane emissions. Based on our data, we hypothesize that the emission of sulfide due to SRB activity in the gypsum treated pond is also limited due to its high solubility and oxidation in surface waters.     

Adsorption of transgenic insecticidal Cry1Ab protein to silica particles. Effects on transport and bioactivity

Bt crops are genetically modified to be resistant against insect pests by expressing insecticidal Cry proteins. The processes governing the fate and bioavailability of the expressed transgenic Cry proteins in soils are poorly understood. We studied adsorption of Cry1Ab to negatively charged silica (SiO(2)) particles, a major soil constituent and a model for negatively charged mineral surfaces, at pH 5 to 10 and ionic strengths I = 10 mM to 250 mM, both in solution depletion and saturated column transport experiments. Cry1Ab-SiO(2) interactions were dominated by patch-controlled electrostatic attraction (PCEA), as evident from increasing Cry1Ab attraction to SiO(2) with decreasing I at pH at which both Cry1Ab and SiO(2) were net negatively charged. Experimental and modeling evidence is provided that the surface heterogeneity of SiO(2) particles modulated PCEA, leading to a fraction of adsorption sites with slow Cry1Ab desorption kinetics. Desorption rates from these sites increased upon increasing the solution pH. In toxicity bioassays, we demonstrated that Cry1Ab retained insecticidal activity when adsorbed to SiO(2), suggesting high protein conformational stability during adsorption-desorption cycles. Models predicting Cry1A protein adsorption in soils therefore need to account for combined effects of the nonuniform protein surface charge distribution and of sorbent surface heterogeneity.