An exploratory study on the use of enzyme activities as sediment tracers: biochemical fingerprints?

Little information exists on the potential of soil enzyme activities,which are sensitive to soil properties and management,for the characterization of sediment sources at the catchment scale.The objective of this study is to explore and evaluate enzyme activity as tracer for sediment fingerprinting in the Hiv catchment(55 km~2),Iran.Therefore,four enzymes were measured from 42 different sampling sites,covering three sediment source areas(rangeland/surface erosion,orchard/surface erosion,and streambank erosion),as well as from 12 sediment samples from reservoir check dams (sediment sinks).The results indicate that,based upon backward mode discriminant analysis,βglucosidase and dehydrogenase,allowed more than 95%of the samples to be correctly assigned to their source areas.These enzymes were selected as input data for a mixing-model to determine the relative contribution of the sampled sediment sources.The mean contributions from rangeland, orchard and streambank sources in the study area were estimated as 11.3%,15.1%and 73.7%, respectively.Using geochemical tracers,the mean contribution from rangeland,orchard and streambank sources was estimated as 14.1%,9.5%and 74.8%,respectively.Combined biochemical and geochemical tracers,similar values were obtained(18.7%,10.7%and 70.7%,respectively).Our results indicate that soil enzyme activity allows for a good characterization of sediment sources,and can provide a complementary tool to currently existing sediment fingerprinting approaches.However, the method should be also tested in other regions.     

Terbuthylazine and Deethylterbuthylazine in Rain and Surface Water – Determination by Enzyme Immunoassay and Gas Chromatography/Mass Spectrometry

Rain and surface water samples from Southern Germany were investigated from 1991 to 1995 for terbuthylazine and one of its major metabolites, deethylterbuthylazine. The concentrations observed were compared to the concentrations found for atrazine and deethylatrazine in the same water samples. Concentrations ranged from < 0.02 μg/L to 0.7 μg/L for terbuthylazine and from < 0.02 μg/L to 0.6 μg/L for deethylterbuthylazine, compared to concentrations of < 0.02 μg/L to 3 μg/L and < 0.02 μg/L to 0.5 μg/L for atrazine and deethylatrazine, respectively. The ratios of metabolite concentrations to parent compound concentrations were calculated for deethylterbuthylazine to terbuthylazine (DTR) and deethylatrazine to atrazine (DAR). In rain water, DTR of 0.8…3.0 and DAR of 0.3… 1.9 were determined with mean values of 0.9… 1.7 for DTR and 0.6…0.9 for DAR in the different years. The ratios increased during summer periods. The highest ratios were observed in samples from forest stands, showing that degradation of the herbicide has occurred during transport between the source and the sampling site. The DTR in rain water were about 50… 100% higher than the DAR. This indicates a higher degradation rate of terbuthylazine during atmospheric transport. In surface water, DTR of 0.3… 1.2 with mean values of 0.5…0.8 and DAR of 0.2…2.2 with mean values of 0.2… 1.3 were observed. The ratios increased from June to September.     

Gewässergütebewertung und Bewertung von Abwasserfahnen in Rhein und Main auf der Basis mikrobieller Enzymaktivitäten

Quality Mapping of Surface Water and Assessment of Treated and Untreated Waste-water Inputs into the Rhine and Main River Based on Microbial Enzyme Activities Microbial enzyme activities are used for an extension of the traditional quality mapping of surface water. In the following study, the enzymatic parameters were proved and validated on samples from various creeks and rivers in Baden-Württemberg and on samples gathered from the Rhine and Main Rivers (Germany). The test parameters should also be used for the control and the preservation of the capacity of the biological self-purification, which is the only natural and essential process in drinking-water conditioning of surface water. Main subject of the surveys was the development of criteria for an assessment of the measured enzymatical inhibition effects. Therefore, classes of inhibition and indices of inhibition are defined.     

Surface associations of enzymes and of organic matter: Consequences for hydrolytic activity and organic matter remineralization in marine systems

The extent to which marine organic matter is associated with surfaces and the consequences of such associations for organic matter remineralization are the focus of considerable attention. Since extracellular enzymes operating outside microbial cells are required to hydrolyze organic macromolecules to sizes sufficiently small for substrate uptake, the effects of surface interactions–on enzymes as well as on substrates–for hydrolytic activity also require investigation. We used a simplified laboratory system consisting of a free (dissolved) polysaccharide (pullulan) and the same polysaccharide tethered to agarose beads to restrict mobility, plus the corresponding free enzyme (pullulanase) and the same enzyme sorbed to montmorillonite (Mte), to investigate systematically the consequences of surface associations of enzymes and of substrates on hydrolytic activity. Changes in substrate molecular weight were monitored with time to measure the course of enzymatic hydrolysis. Although hydrolysis of free substrate was nearly complete after 2 min incubation with the free enzymes, the sorbed enzymes also effectively hydrolyzed free substrate, and the data suggest that they retained activity longer in solution compared to the free enzymes. Sorbed enzymes progressively hydrolyzed the free substrate from > 50 kD to lower molecular weights during a 24 h incubation, with a final product distribution on average showing only 1.4% and 10.3% of substrate still in the > 50 kD and 10 kD size classes, while 46.6%, 29.3%, and 12.5% of substrate was in the 4 kD, monomer, and free tag size classes, respectively. This product distribution was very similar to that of the free substrate/free enzyme experiment. Tethering the substrate to agarose beads led to lower substrate release (2–3% of total substrate after 98 h incubation) into solution compared to the free substrate case. For tethered substrates, the state of the enzyme (free or sorbed) measurably affected the molecular weight distribution of the hydrolysis products, with free enzymes producing a higher fraction of high molecular weight hydrolysis products (28.7 ± 5.4% of substrate > 50 kD at the end of the incubation) compared to sorbed enzymes (11.6 ± 2.8% of substrate > 50 kD at the end of the incubation.) Tethered substrates were also hydrolyzed in a sediment slurry from surface sediments from Cape Lookout Bight, North Carolina; 0.1% of total substrate was released by enzymes naturally present in 1 cm³ of sediment after 144 h incubation, demonstrating that the enzymes naturally present in marine sediments are also capable of accessing tethered substrates. These investigations suggest that surface associations of enzymes in marine systems may extend the active lifetime of such enzymes, providing an opportunity for hydrolysis over longer periods of time and producing a different size spectrum of hydrolysis products relative to free enzymes. Furthermore, in well-mixed systems, surface-associated enzymes can hydrolyze substrates whose mobility is restricted, highlighting the importance of processes such as resuspension and bioturbation on organic matter remineralization.     

A comparative analysis of enzyme leach and mobile metal ion selective extractions; case studies from glaciated terrain, northern Ontario

The enzyme leach and MMI (mobile metal ion) selective digestions are rapidly gaining popularity in the exploration community because of their successful track record in non-glaciated, arid and tropical climatic regions. Few successful case studies from high-latitude, temperate regions have been published to date. Also lacking is a comprehensive discussion of the constraints and limitations of these geochemical methods and how they affect the successful interpretation of selective-leach survey results in glaciated regions. This paper evaluates each method using specific examples from four case studies undertaken in the glaciated region of northern Ontario. The case studies clearly demonstrate that both the MMI and enzyme leach selective digestions provide the explorationist with useful information, which, when used in conjunction with other exploration tools, can assist with the identification of potential diamond drill targets. Notwithstanding this, the techniques do not appear to work well as ‘stand alone' methods. The importance of following strict sampling protocols and obtaining high-quality observational information on the sampling site and media collected cannot be overstressed. It is only by filtering out the many variables that arise in any sampling program that a sound interpretation of the data can be made. The case studies have drawn attention to some of the apparent shortfalls of the techniques. Of these, issues such as the ability to reproduce survey results from year to year and the recognition of anomalous element associations which specifically target blind mineralisation are probably those which require significant follow-up work. Clearly, additional case studies are required from the glaciated regions of the world to assist with the optimisation of geochemical response of MMI and enzyme leach selective extractions. The mechanisms responsible for the formation of surface geochemical anomalies over deeply buried mineralisation are not well understood. The observed patterns of geochemical response in surface soils and both shallow and deep ground waters over concealed mineralisation should assist with the development of new dispersion models.     

合浦珠母贝碱性磷酸酶的分离纯化与性质研究

以合浦珠母贝（Ｐｉｎｃｔａｄａ ｆｕｃａｔａ）为提取酶的原料,经Ｔｒｉｓ－ＨＣｌ缓冲液（ｐＨ７．５）抽提,正丁醇处理,硫酸铵分级沉淀分离,ＤＥＡＥ－纤维素离子交换树脂支析,ＤＥＡＥ－Ａ２５离子交换分子筛柱层析纯化,得到一定纯度的碱性磷酸酶,比活力达到１４４４Ｕ／ｍｇ。酶学性质和动力学性质研究表明,该酶催化对硝基苯磷酸（ｐＮＰＰ）的水解反应,最适ｐＨ值为９．７２,最适温度为４５℃,水解反应的活化能为     

Enzyme Immunoassay for Pesticide Analysis Enzymimmunoassays zur Pestizidanalytik

Enzyme immunoassays profit from the unique properties of antibodies concerning their selectivity and affinity towards ligands, e.g. pesticides. The most commonly applied types belong to the class of competitive immunoassays with phase separation such as the ELISA. The principle is explained and illustrated using an atrazine assay as a representative example. Common error sources, especially cross-reactivities and matrix effects, are discussed. Interlaboratory tests demonstrate the presently available quality of enzyme immunoassays. An outlook is given on future developments.     

Approaches to the Immunochemical Analysis of Non-extractable Triazine Residues in Refractory Organic Substances (ROS) and Characterization of ROS

The tendency of several pesticides to form non-extractable residues requires simple and fast screening methods. Immunochemical methods can meet this demand. They are based on selective and highly sensitive antibody binding to pesticide residues exposed on refractory organic substances (ROS). s-Triazines were used as an example to show the feasibility of this approach. It was proven that under laboratory conditions atrazine was bound to ROS. Antibody recognition pointed to the availability of free ethyl and isopropyl groups exposed at the non-extractable s-triazine residue. Therefore, it is suggested on the basis of antibody selectivities that binding of atrazine to ROS may take place by substitution of the chlorine residue. In the next step, aquatic ROS isolated from surface waters located in an agricultural area were examined for non-extractable s-triazines. ROS from a lake were found to contain bound residues, whereas none were detected in the creek samples. As an outlook the feasibility of serological characterization of ROS is discussed. The novel concept of serotyping is based on the fractionation of ROS by electrophoretic techniques followed by an immunoreaction with polyclonal antibodies directed against different ROS. When immunoelectrophoresis is used, precipitation lines can be compared and checked for identity and cross-reactivity. This approach can form the basis for classifying ROS and determine similarities with unknown ROS.     

Structure and function of soil microbial community in artificially planted Sonneratia apetala and S. caseolaris forests at different stand ages in Shenzhen Bay,China

The present study examined the relationships between soil characteristics, microbial community structure and function in the forests artificially planted with exotic Sonneratia apetala at stand ages of 1-, 2-, 7-, 10- and 14-years and Sonneratia caseolaris of 1-, 4-, 7-, 10- and 14-years in Futian National Nature Reserve, Shenzhen Bay, China. The 7-years old forests of both Sonneratia species reached peak growth and had the highest content of nitrogen and phosphorus, enzymatic activities, including dehydrogenase, cellulase, phosphatase, urease and ß-glucosidase, except arylsulphatase which increased continuously with stand ages. The microbial community structure reflected by phospholipid fatty acid (PLFA) profiles also reached the maximum value in the 7-years old forests and soil bacterial PLFAs in both forests were significantly higher than fungal PLFAs. The canonical correlation analysis revealed that differences in microbial structural variables were significantly correlated to the differences in their functional variables, and the highest correlation was found between the soil enzymatic activities and the content of carbon and nitrogen.