Die Böden von Wallhecken in Schleswig-Holstein. II. Aufbau und Umsetzung der organischen Bodensubstanz

Soils of Mound Hedges in Schleswig-Holstein. II. Composition and Transformation of Soil Organic Matter Two Cumulic Anthrosols of mound hedges and the fossil A-horizons below the deposits, were investigated by means of wet chemistry and ¹³C-NMR spectroscopy in order to characterize soil organic matter (SOM) and to compare their SOM with natural and recent soils. The bulk humus of the Cumulic Anthrosols contains a high amount of easily soluble polysaccharides. Despite their high bioavailability sugars, starch and mobile fulvic acids are not completely consumed by microorganisms and translocated into the subsoil. The initial phase of podzolization is indicated due to this process and the very low pH-level of the soils. After deposition about 30 % of the soil carbon in the A-horizons is mineralized. Litter compounds are predominantly decomposed, whereas humic compounds are selectively preserved or newly synthesized. The significance of the humic compounds in the soil organic matter increases after deposition. This is why the potential cation exchange capacity of the fossil Ahorizons is similar to this of the recent ones.     

Die Standortsbewertung der biologischen Aktivität von Böden über Ermittlung der Bodenatmung und der zellulolytischen Aktivitatät im Feld

Intersite estimation of biological activity of soils by determination respiration and cellulolytical activity in the field A temperature differentiated scale of estimation is presented, which is derived from long-term investigations of respiration and cellulolytic activity. This is suitable for intersite comparison of different soil-ecosystems. Investigations in spring are representative and could be done during mapping the soil-ecosystems. It exists a strong correlation to the quality of the humus body and to the available nutrients.     

Comparative genotoxicity testing of rhine river sediment extracts using the comet assay with permanent fish cell lines (rtg-2 and rtl-w1) and the ames test*

Goals, Scope and Background

Improved quality of surface waters and sediments requires advanced strategies for ecotoxicological assessment. Whilst at least in Germany assessment strategies on the basis of chemical analysis and acute toxicity data dominated the last decades, the development of more specific biological endpoints and biomarkers in ecotoxicology is required in order to arrive at a good ecological potential and good chemical status of surface waters in the European river basins until the year 2015, as required by the European Water Framework Directive. Since sediments have for long been known to function both as a sink and as a source of pollutants in aquatic systems, and since part of the particle-associated substances have frequently been demonstrated to cause mutagenic and carcinogenic effects in aquatic organisms, particularly in fish, there is, among other requirements, an urgent need to develop, standardize and implement integrated vertebrate-based test systems addressing genotoxicity into recent sediment investigation strategies. Thus, the present study was designed to compare the suitability of two commonly used test systems, the comet assay and the Ames test, for the evaluation of the ecotoxicological burden of surface and core sediment samples from the river Rhine.

Methods (or Main Features)

In order to determine the importance of inherent enzymatic activities, two permanent fish cell lines with different biotransformation capacities, RTL-W1 and RTG-2, were compared with respect to their capability of detecting genotoxic effects in 18 surface and core sediment samples from 9 locations along the River Rhine in the comet assay with and without exogenous bioactivation. For further comparison, as a prokaryotic mutagenicity assay, theSalmonella plate incorporation assay (Ames test) with the test strains TA98 and TA 100 with and without exogenous metabolic activation was used.

Results and Discussion

Whereas all sediment extracts induced genotoxic effects in the comet assay with RTL-W1 cells, only 12 out of 18 sediment extracts revealed significant genotoxicity in the tests with the less biotransformation-competent RTG-2 cells. Exogenous bioactivation by addition of ß-naphthoflavone /phenobarbital-induced S9 from rat liver resulted in both reduction or increase of genotoxicity in samples from different sites, however, without consistent reaction patterns. In general, the responses of RTL-W1 cells indicated higher biotransformation capacity than in RTG-2 cells without S9 complementation. In Ames tests using TA98 with S9, 16 out of 18 extracts induced significant mutagenicity with induction factors up to 4. Compared to TA98, the strain TA100 proved less sensitive, with maximum induction factors of 1.3, indicating the potential presence of substances inducing frarneshift mutations, which can only be detected in the strain TA98. Chemical analyses revealed particularly high levels of hexachlorbenzene (up to 860 µg/kg) and priority PAHs (up to 4.8 mg/kg); so far, however, no correlation could be found between compounds analyzed and the corresponding biotests.

Conclusions

Results document that both comet assay and Ames test are capable of detecting xenobiotic interaction with DNA in consequence of exposure to complex environmental samples. Whereas the alkaline version of the comet assay detects a broad range of interactions with the DNA, however without information about their eventual importance, the Ames test only reveals established mutations, but fails to detect transient (reparable) DNA alterations. However, even transient primary changes in the DNA structure might result in carcinogenic processes and, eventually, in implications at the population level. As a consequence, for hazard assessment purposes, a combination of both assays is required to avoid false negatives in genotoxicity evaluation. Poor correlation between data obtained by chemical analysis and results in bioassays is indicative of our limited understanding of the sources of genotoxicity. In fact, numerous studies combining chemical and biological approaches for hazard assessment of complex environmental mixtures indicate that priority pollutant concentrations are a poor indicator of toxicity.If compared to the cell line RTG-2, RTL-W1 proved more effective in detecting genotoxicity in surface sediment samples and, thus, indicated the importance of bioactivation of at least part of the compounds in superficial layers of sediments. Results further document that the common assumption may be wrong that, in comparison to deeper strata, surface layers carry a lower toxic burden in consequence of the current decrease in water pollution. This might at least in part be due to remobilization of more heavily polluted sediments from deeper layers during severe flood events followed by re-sedimentation in flood plains or upstream weirs, where they might cover less polluted younger sediment layers.

Recommendations and Perspectives

For a comprehensive assessment of genotoxicity in surface and core sediments, a combination of eukaryotic (comet assay) and prokaryotic assays (Ames test) with and without exogenous bioactivation is recommended. Since studies with organic sediments extracts simulate a worst-case scenario and fail to take into account bioavailability, there is broad consensus that whole-sediment exposure protocols represent the most realistic scenarios. Whereas more realistic solid phase exposure has frequently been applied in both microbial and invertebrate acute toxicity testing, there is an urgent need to develop corresponding whole sediment fish-based genotoxicity tests.

     

Structure-antioxidant efficiency relationships of phenolic compounds and their contribution to the antioxidant activity of sea buckthorn juice

The phenolic composition of juice derived from fruits of sea buckthorn (Hippophae rhamnoides) was investigated by high-performance liquid chromatography (HPLC) with diode array and electrochemical detection. Flavonols were found to be the predominating polyphenols while phenolic acids and catechins represent minor components. Of the seven flavonols identified, isorhamnetin 3-O-glycosides were the most important representatives quantitatively. However, because of their structural properties, they were poor radical scavengers as shown by electron spin resonance spectroscopy. Phenolic compounds such as quercetin 3-O-glycosides, catechins, and hydroxybenzoic acids with a catechol structure exhibited good antioxidant capacities, but their concentration in sea buckthorn juice was small. These phenolic compounds, determined by HPLC, accounted for less than 5% of the total antioxidant activity of the filtered juice. Ascorbic acid was shown to be the major antioxidant in sea buckthorn juice. Because of its high concentration of 1.22 g/L, it contributes approximately 75% to total antioxidant activity. The remaining difference can be attributed to higher molecular weight flavan-3-ols (proanthocyanidins), which were determined photometrically after acid depolymerization to colored anthocyanidins.     

Comparing the results of the serological detection of Salmonella antibodies in blood serum and meat juice from different muscles from slaughter pigs

The German Salmonella Monitoring Programme started by the QS-System in 2002 (Blaha, 2004) is mandatory due to the so-called "Salmonella Regulation for Pigs" since 2007 (Anonym, 2007). The Regulation does not clearly prescribe the specific muscle which is to be taken as source of the meat juice. Thus, at different slaughter plants meat samples are also taken from different muscles and several scientific papers describe various muscles as source of the meat juice, too. The objective of this study was to compare the serological results of meat juices from three different locations (diaphragm pillar, neck, belly muscle) to each other and to those of the blood serum from exactly the same animals. All samples were simultaneously tested for Salmonella antibodies by two serological tests (Salmo-type Pig Screen, LDL, Germany; HerdChek Swine Salmonella, IDEXX, Germany). Comparisons were carried out between the various sample kinds per animal and between the two test systems. The analysis of all results of the meat juices revealed in both test systems a clear decline of the OD% values from the diaphragm pillar to the neck to the belly muscle. The average OD% values of all samples were higher when measured by the HerdChek ELISA (IDEXX, Germany) than by the Salmotype ELISA (LDL, Germany), especially in blood serum. Since the results of the meat juice samples gained from the diaphragm pillar were in both test systems by far the closest to the results of the corresponding serum blood samples, it is recommended to amend the "Salmonella Regulation for Pigs" by prescribing meat from the diaphragm pillar as the only muscle for gaining meat juice.     

Bayesian space-time analysis of Echinococcus multilocularis-infections in foxes

A total of 26,220 foxes that were hunted or found dead in Thuringia, Germany, between 1990 and 2009 were examined for infection with Echinococcus multilocularis, the causative agent of human alveolar echinococcosis, and 6853 animals were found infected. The available data on the foxes including the location (local community; district) and the date of hunting/death were analyzed using a hierarchical Bayesian space-time model. The distribution of the model parameters and their variability was estimated on the basis of the sample size, the number of cases per spatial unit and time interval, and an adjacency matrix of the municipalities using a Markov Chain Monte Carlo simulation technique to assess the spatial and temporal changes in the distribution of the parasite. The model used to evaluate the data is widely applicable and can be applied to analyse data sets with gaps and variable sample sizes per spatial and temporal unit. In the study area, the prevalence of E. multilocularis increased from 11.9% (95% confidence interval 9.9-14.0%) in 1990 to 42.0% (39.1-44.1%) in 2005. While the infection was present in foxes only in the north-western parts of Thuringia in 1990, it had spread over the entire state by 2004. These results demand increased vigilance for human alveolar echinococcosis in Thuringia.     

Clinical evaluation of the CA530-VET hematology analyzer for use in veterinary practice

BACKGROUND: The CA530-VET is a completely automated impedance cell hematology analyzer, which yields a 16-parameter blood count including a 3-part leukocyte differential. OBJECTIVES: The aim of this study was to examine the operational potential of the CA530-VET and its value for use in veterinary practice. METHODS: The analyzer was tested for blood carry-over, precision, and accuracy. Comparison methods included the CELL-DYN 3500, microhematocrit centrifugation, manual platelet (PLT) counting for feline and equine species, and a 100-cell manual WBC differential. Blood samples for comparison of the methods were obtained from 242 dogs, 166 cats, and 144 horses. RESULTS: The carry-over ratio (K) was 0.28% for RBC, 0.59% for PLT, 0.32% for WBC, and 0.18% for hemoglobin (HGB) concentration. Coefficients of variation (CVs) for within-batch precision and duplicate measurement of blood samples were clearly within the required limits, except for duplicate platelet counts in cats (8.7%) and horses (9.5%). The WBC count was in excellent agreement for dogs and horses and RBC count was in excellent agreement for horses. The accuracy of feline WBC counts was not acceptable, with the exception of values at the high end of the range. RBC counts in dogs and cats, and HGB concentration and MCV in all 3 species were sufficiently accurate. The CA530-VET HCT results were in excellent agreement with microhematocrit results in horses but exceeded the maximum allowed inaccuracy for cats and dogs. In all species, PLT counts established mechanically and manually were not in adequate agreement. Large differences were found between the CA530-VET and the manual differential percentage for lymphocytes and "mid-sized cells" (monocytes and basophilic granulocytes). CONCLUSIONS: The CA530-VET can be considered useful for routine canine, feline, and equine blood cell analyses. It should not be considered accurate, however, for PLT counts, feline total WBC counts in the subnormal and normal range, and leukocyte differentials, except for granulocytes.     

Antimicrobial susceptibility of Klebsiella spp. and Proteus spp. from various organ systems of horses, dogs and cats as determined in the BfT-GermVet monitoring program 2004-2006

A total of 120 isolates of Klebsiella spp. and Proteus spp. collected from horses and small animals (dogs and cats) were screened for their susceptibility to 24 different antimicrobial agents. Klebsiella spp. were included from infections of the genital tract (GT) of horses (36 isolates) and the urinary/genital tract (UGT) from dogs and cats (17 isolates), while Proteus spp. were from small animal (dogs and cats) infections of the UGT (37 strains) and the skin (incl. ear/mouth) (30 isolates). In Klebsiella spp. resistance appeared most frequently to ampicillin (53-67%), sulfamethoxazole (19-29%) and potentiated sulfonamides (trimethoprim/sulfamethoxazole 1/19 combination) (19-24%). A further 29% of enrofloxacin resistant Klebsiella isolates were observed for the UGT of small animals. From the GT of horses for this antimicrobial agent there was no isolate detected with a comparably high minimum inhibitory concentration (MIC) value. In Proteus spp. highest percentages of resistance occurred against tetracycline (90-92%). Due to drug efflux proteins, high MIC values against this antimicrobial agent have been frequently reported in literature. In Proteus spp. relevant resistance percentages also occurred for potentiated sulfonamides (27-37%), sulfamethoxazole (24-37%) and chloramphenicol (24-37%).     

Cortical constriction during abscission involves helices of ESCRT-III-dependent filaments

After partitioning of cytoplasmic contents by cleavage furrow ingression, animal cells remain connected by an intercellular bridge, which subsequently splits by abscission. Here, we examined intermediate stages of abscission in human cells by using live imaging, three-dimensional structured illumination microscopy, and electron tomography. We identified helices of 17-nanometer-diameter filaments, which narrowed the cortex of the intercellular bridge to a single stalk. The endosomal sorting complex required for transport (ESCRT)-III co-localized with constriction zones and was required for assembly of 17-nanometer-diameter filaments. Simultaneous spastin-mediated removal of underlying microtubules enabled full constriction at the abscission site. The identification of contractile filament helices at the intercellular bridge has broad implications for the understanding of cell division and of ESCRT-III-mediated fission of large membrane structures.