Quantification of bacterial lipopolysaccharides (endotoxin) by GC-MS determination of 3-hydroxy fatty acids

A GC-MS method for the quantification of bacterial lipopolysaccharides (LPS, endotoxin) is presented. After hydrolytic cleavage of 3-hydroxy fatty acids (3-OH FAs) from the lipid A region of LPS, derivatisation of both the hydroxyl and the carboxyl group was performed in one step with a mixture of methyl-bis(trifluoracetamide)(MBTFA) and N-methyl-N-(tert-butyldimethylsilyl)trifluoracetamide (MTBSTFA). Using GC-MS in the EI mode with selected ion monitoring (SIM) for analysis, baseline separation of 3-OH FAs (and of possibly interfering 2-OH FAs) was achieved. The sensitivity of the method (LOD 7-50 pg/injection for the different 3-OH FAs investigated) allows for the efficient quantification of LPS in occupational and environmental samples. Degradation of 3-OH FAs as well as of their derivatives during sample preparation and GC-MS separation as a possible source of errors in analytical methods based on 3-OH FA determination is reported for the first time. Thermal elimination of water from the underivatised 3-OH FAs and of trifluoroacetic acid from the derivatives was identified as the cause of degradation. The resulting alpha,beta-unsaturated compounds showing the same mass spectra as the 3-OH FA derivatives were detected as more or less prominent satellite peaks. By using alkaline instead of acidic hydrolysis and cool on-column instead of split/splitless injection, elimination was reduced to an acceptable level.     

Environmental Monotoring and Assessment Program: Current Status of Virginian Province (U.S.) Estuaries

Monitoring of indicators of the ecological condition of bays, tidal rivers, and estuaries within the Virginian Biogeographic Province (Cape Cod, Massachusetts to Cape Henry, Virginia) was conducted annually by the U.S. EPA's Environmental Monitoring and Assessment Program (EMAP) during the summer months of 1990 to 1993. Data were collected at 425 probability-based stations within the Province. Indicators monitored included water quality (temperature, salinity, water clarity, and dissolved oxygen concentration), sediment contamination, sediment toxicity, benthic community structure, fish community structure, and fish gross external pathology. Data were used to estimate the current status of the ecological condition of Virginian Province estuarine resources, and provide a baseline for identifying possible future changes. Estimates, with 95% confidence limits, of the areal extent of impacted resources within the Province are provided. Data are also presented by estuarine class (large estuaries, small estuarine systems, and large tidal rivers). Results show that, overall, approximately half of the estuarine waters of the Virginian Province can be classified as impacted based on multiple indicators, with hypoxia being the major stressor.     

Kosovo-Konflikt: ?kologische Auswirkungen

Zusammenfassung  Der WWF (World Wide Fund for Nature), Initiator und Tr?ger des Donau-Karpaten-Programms, besch?ftigt sich seit Beginn des Kosovo-Konfliktes mit den Auswirkungen der kriegerischen Auseinandersetzungen auf Natur und Mensch und versucht, die Welt?ffentiichkeit entsprechend zu informieren. Ende Juli 1999 hat der WWF International eine Mission, bestehend aus unabh?ngigen Toxikologinnen und ?lexperten, an die Donau entsandt. Unter der Leitung von Philip Weller, dem Direktor des WWF Donau-Karpaten-Programms, wurden drei Tag lang Industriekomplexe besichtigt und Gespr?che mit Betroffenen geführt. Die Reise führte das Team nach Panćevo und Novi Sad, und damit zu den Schwerpunkten der zerst?rten Chemie- und Petrochemischen Komplexe. In den betroffenen Werken, HIP AZOTARA, Panćevo und NIS ?lraffinerie, Novi Sad wurde das Team von den jeweiligen Werkleitern geführt. Gespr?che fanden statt mit dem Bürgermeister von Panćevo und Vertretern des Gesundheitsamtes in Belgrad. Es war dem Team m?glich, Boden- und Wasserproben zu nehmen und zu analysieren. Damit liegen den Autoren unabh?ngige Analysenergebnisse vor, die es gestatten, eine erste, orientierende Einsch?tzung der Kontaminationen vorzunehmen.      

Enzymatic production of biosilica glass using enzymes from sponges: basic aspects and application in nanobiotechnology (material sciences and medicine)

Biomineralization, biosilicification in particular (i.e. the formation of biogenic silica, SiO₂), has become an exciting source of inspiration for the development of novel bionic approaches following “nature as model”. Siliceous sponges are unique among silica forming organisms in their ability to catalyze silica formation using a specific enzyme termed silicatein. In this study, we review the present state of knowledge on silicatein-mediated “biosilica” formation in marine sponges, the involvement of further molecules in silica metabolism and their potential application in nanobiotechnology and medicine. Werner E. G. Müller dedicated this study to Prof. Vera Gamulin (Rudjer Boskovic Institute, Zagreb, Croatia) in honour of her unique contributions in molecular evolution.     

Bewertungskriterien des Umweltbundesamtes: Auswirkungen von Pflanzenschutzmitteln auf terrestrische Arthropoden

The fundamental data requirements for the authorization of plant protection products and the inclusion of active ingredients in Annex I of Council Directive 91/414/EEC are described in the Annexes II and III of the Directive. Definite guidelines concerning investigations with arthropods are partially deficient. The uniform principles for the registration of plant protection products in the Member States described in Annex VI of the directive were primarily developed to assess the effects on the beneficial capacity of arthropods in agriculture. However, the risk for arthopods as an essential part of the environment need to be assessed, too. Hence, the German Federal Environmental Agency has developed a tiered approach to assess the effects of plant protection products on arthopods based on the data requirements of Annex II and III of the Directive 91/414/EEC. The risk is assessed using the effect-concentration evaluated in ecotoxicological tests and the environmental concentration predicted by validated exposure models. To protect non-target species in terrestrial ecosystems assessment factors need to be considered.     

Rapid tissue reduction and recovery in the sponge Aplysinella sp.

We observed a pronounced, yet reversible tissue reduction in the tropical sponge Aplysinella sp. under non-experimental conditions in its natural habitat, after transfer into seawater tanks, as well as after transplantation from deep to shallow water in the field. Tissue reduction resulted in the formation of small “reduction bodies” tightly attached to the sponge skeleton. Although volume loss and gain were substantial, both tissue reduction and regeneration were often remarkably rapid, occurring within few hours. Microscopic analysis of the reduction bodies revealed morphological similarities to previously described sponge primmorphs, with densely packed archaeocytes and spherulous cells enclosed by a thin layer of epithelial-like cells. Denaturing gradient gel electrophoresis (DGGE) revealed pronounced changes in the sponge-associated microbial community upon tissue reduction during laboratory and field experiments and following changes in ambient conditions after transplantation in the field. Generally, the microbial community associated with this sponge proved less stable, less abundant, and less diverse than those of other, previously investigated Verongid sponges. However, one single phylotype was consistently present in DGGE profiles of Aplysinella sp. This phylotype clustered with γ-proteobacterial sequences found previously in other sponge species of different taxonomic affiliations and geographic provenances, as well as in sponge larvae. No apparent changes in the total secondary metabolite content (per dry weight) occurred in Aplysinella sp. upon tissue reduction; however, comparative analysis of intact and reduced tissue suggested changes in the concentrations of two minor compounds. Besides being ecologically interesting, the tissue reduction phenomenon in Aplysinella sp. provides an experimentally manipulable system for studies on sponge/microbe symbioses. Moreover, it may prove useful as a model system to investigate molecular mechanisms of basic Metazoan traits in vivo, complementing the in vitro sponge primmorph system currently used in this context.     

SNOTEL,the Soil Climate Analysis Network,and water supply forecasting at the Natural Resources Conservation Service: Past,present, and future

The Snow Survey and Water Supply Forecasting (SSWSF) Program and the Soil Climate Analysis Network (SCAN) of the United States Department of Agriculture's Natural Resources Conservation Service (NRCS) generate key observational and predictive information for water managers. Examples include mountain climate and snow monitoring through manual snow surveys and the SNOw TELemetry (SNOTEL) and SNOtel LITE networks, in situ soil moisture data acquisition through the SCAN and SNOTEL networks, and water supply forecasting using river runoff prediction models. The SSWSF Program has advanced continuously over the decades and is a major source of valuable water management information across the western United States, and the SCAN network supports agricultural and other water users nationwide. Product users and their management goals are diverse, and use-cases range from guiding crop selection to seasonal flood risk assessment, drought monitoring and prediction, avalanche and fire prediction, hydropower optimization, tracking climate variability and change, environmental management, satisfying international treaty and domestic legal requirements, and more. Priorities going forward are to continue innovating to enhance the accuracy and completeness of the observational and model-generated data products these programs deliver, including expanded synergies with the remote sensing community and uptake of artificial intelligence while maintaining long-term operational reliability and consistency at scale.     

Comparing growth development of <Emphasis Type="Italic">Myriophyllum</Emphasis> spp. in laboratory and field experiments for ecotoxicological testing

BACKGROUND, GOALS AND SCOPE: Risk assessment of herbicides and the evaluation of contaminated sediments based on algae and the macrophyte Lemna sp. alone may underestimate the potential hazard of certain compounds. Therefore, various test systems with Myriophyllum spp. have been developed recently to assess the phytotoxicity in surface waters and natural sediments. In the present study, experiments investigating the growth development of Myriophyllum spp. were performed in the laboratory under defined conditions and in mesocosms under environmentally realistic exposure conditions to evaluate the suitability of these species as potential standard test organisms in ecotoxicological testing. This study provides data on the endpoints biomass, plant length and root development. MATERIALS AND METHODS: Six independent experiments were performed to investigate the plant development of Myriophyllum spp. under control conditions. The main difference in the experiments was the complexity of the test systems ranging from simple laboratory experiments to complex outdoor mesocosm studies. At the start of each experiment, uniform cuttings of Myriophyllum spp. were placed in vessels with or without sediments to reduce variability between replicates. The endpoints considered in this investigation were biomass (fresh weight of the whole plant), length of the main shoot, length of the side shoots, total length of the plant (calculated from the length of the main and side shoots) and root formation. Root to shoot ratios were calculated as a further measure for plant development. Relative growth rates (RGR) based on plant length (RG(L)R) and on biomass (RG(B)R) were calculated. RESULTS: Despite the various experimental conditions, comparable growth was obtained in all test systems and the variability of endpoints, such as total length and biomass of plants, was low. It was observed that the RGR of M. spicatum in the simple laboratory test system with sediment were comparable to growth data obtained for M. verticillatum and M. spicatum grown in indoor and outdoor mesocosms, thus indicating that Myriophyllum growth tends to increase by the addition of sediment. High variability was determined for the endpoints length of the side shoots, total root length and biomass of roots. DISCUSSION: One challenge for a test design to investigate phytotoxicity on aquatic plants is to obtain good growth of the plants. From the results, it can be concluded that the experimental conditions in the various test systems were suitable to study the plant development of Myriophyllum spp. because obtained growth rates were comparable between laboratory and field investigations. Another challenge for developing a plant biotest system is the definition of sensitive endpoints. Low variability is preferred to detect minor effects of chemicals or polluted sediments on plant development. In our studies, the variability of the endpoints biomass and total length of plant was low and, therefore, they have much potential as endpoints for assessing toxicity. CONCLUSIONS: The methodologies presented in this study have applications within the risk assessment for aquatic plants and have the advantage of assessing effects taking into account the relevant exposure pathways via water and/or sediment for compounds under investigation. RECOMMENDATIONS AND PERSPECTIVES: Setting safe quality criteria for surface water and sediments is one of the challenges authorities are facing today. Myriophyllum spp. is recommended as suitable test species to investigate phytotoxicity in surface water and sediments. These results, thus, might serve as a basis for the compilation of a new harmonised guideline for ecotoxicological testing with aquatic macrophytes.     

Gaseous Emissions from Vehicles in a Traffic Tunnel in Vancouver,British Columbia

ABSTRACT

In August 1995, measurements of CO, NO_x, speciated nonmethane hydrocarbons (NMHC), and CO₂ were made in Vancouver's Cassiar Connector, a 730-m-long level-grade highway traffic tunnel. Two characteristics of the Vancouver setting are the presence of many propane vehicles and a mandatory inspection and maintenance (I/M) program. Although the driving conditions and vehicle fleets are otherwise outwardly similar to those of recent Tuscarora-tunnel studies, CO/NO ratios at the Cassiar Connector are significantly lower than those measured at Tuscarora. The Cassiar measurements are consistent with the MOBILE5A mobile emissions model predictions. The Canadian version of MOBILE5A—known as MOBILE5C—gives nearly identical results, indicating that differences in Canadian and U.S. emission standards cannot explain differences between Cassiar and U.S. tunnels. Considering the modeling results as well as measured ethene/acetylene ratios indicative of noncatalyst vehicles, it appears that vehicle deterioration remains the major issue in in-use vehicle emissions—even in Vancouver, where there is a mandatory loaded-mode I/M program.     

Untersuchung von Abfällen mit biologischen Testverfahren zur Bewertung der Wassergefährdung

Wastes from three different types of waste treatment facilities (slag from a municipal waste incineration plant, slag granules from a pilot plant combining carbonization and incineration, mechanical and biological treated wastes) were examined to determine their hazard potential to different waters sites. The process temperature is seen to be the main difference between the three treatment processes. The wastes were extracted with water according to the German standard DIN 38414 S 4 and additionally at a constant pH value of 4. The leachates were investigated in a battery of aquatic bioassays and characterised physically and chemically. Every leachate revealed in a toxic effect at least in one test. The toxicity of the leachates prepared at a pH of 4 was significantly higher than the toxicity of the leachates prepared by extraction with water without pH adjustment. The leachates of the slag granules showed the lowest toxicity. On the basis of these experimental results, a scheme to derive Water Hazard Classes of wastes, which is presented in part II of this publication, was developed.