Bioassays for the ecotoxicological and genotoxicological assessment of contaminated soils (Results of a round-robin test)

For the assessment of contaminated or remediated soils, aside from chemical analyses, ecotoxicological tests are performed which focus on the retention function of soils (determined by tests with aqueous soil extracts) and on the habitat function (determined by tests with soil). Whereas numerous tests exist as standardized guidelines for identifying the effect of chemicals, this is not the case for the assessment of soil quality. A round-robin test was performed to monitor the comparability of the results from ecotoxicological test methods on soils and to facilitate the standardization of corresponding test methods. Four contaminated soils were tested using a total number of fifteen test systems, including ecotoxicological and genotoxicological tests with soil extracts and soil. In the second part of this publication series, the results obtained from the tests with soil microorganisms and soil fauna are presented.     

Does H+ pumping by plasmalemma ATPase limit leaf growth of maize (Zea mays) during the first phase of salt stress?

In the first phase of salt stress, growth of plants is impaired mainly by osmotic stress. To elucidate the effect of NaCl salinity on elongation growth of maize leaves in the first phase of salt stress, we investigated the effect of NaCl on gene expression and activity of the plasmalemma H⁺ ATPase of elongating leaves of maize (Zea mays L.). Treatment of maize plants with 125 mM NaCl for 3 d decreased leaf growth relative to control plants (1 mM NaCl). Whereas H⁺ ATPase hydrolytic activity was unaffected, the ability of the H⁺ ATPase to establish a pH gradient was strongly reduced. Total mRNA of plasmalemma H⁺ ATPase was slightly increased. However, mRNA of the ATPase isoform MHA1 was significantly reduced and ATPase isoform MHA4 was strongly increased at the mRNA level. Synthesis of total H⁺ ATPase protein was unchanged as revealed by western blot. The results indicate that reduced pumping of H⁺ ATPase in leaf plasmalemma under salt stress may be caused by a switch to gene expression of the specific isoform MHA4, which shows inferior H⁺‐pumping efficiency in comparison to isoforms expressed under control conditions. We propose that reduced H⁺ pumping of plasmalemma H⁺ ATPase is involved in the reduction of leaf growth of maize during the first phase of salt stress.     

Effect of different soil cultivation systems,including no-tillage,on electro-ultrafiltration extractable organic nitrogen

The effect of different long-term soil-cultivation systems (ploughing, two types of cultivator, notillage) on organic N, extracted with the electroultrafiltration (EUF) technique, was studied in two arable soils, a Luvic Phaeozem derived from loess and a Eutric Cambisol. A modified EUF extraction procedure (the 80°C fraction extended from 5 to 90 min) was used to investigate the release of organic N from the ploughing and no-tillage treatment, and the content of hydrolysable N was measured in the combined filtrates. No-tillage and the two non-turning cultivation systems led to an accumulation of EUF organic N in the 0- to 10-cm depth compared to the ploughing treatment. In the lower horizon (15–25 cm) the reverse pattern was found in the loamy soil, with higher concentrations after ploughing than after reduced tillage. However, in the sandy soil all four cultivation treatments showed similar values in the 15- to 25-cm depth. During the time of investigation (May 1987 to February 1989) an EUF organic N accumulation occurred, which was about twice as high in the loamy as in the sandy soil. Therefore we conclude that in the sandy soil the mineralization of organic N was faster, and that reduced tillage retarded its degradation. In the total 0- to 25-cm depth, this delay was not observed in the loamy soil. The N release rates were much lower in the sandy than in the loamy soil and they were higher for the notillage than for the ploughing treatment. Only 30–40% of the total organic N desorbed was hydrolysable and the amino acid composition indicates that part of it originated from microbial Cells. The overall evaluation showed clearly that EUF-extractable organic N is a sitespecific factor.     

Determination of 12 type A and B trichothecenes in cereals by liquid chromatography-electrospray ionization tandem mass spectrometry

A new sensitive method for the simultaneous determination of 12 trichothecenes (deoxynivalenol, nivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, fusarenon X, T-2 toxin, HT-2 toxin, neosolaniol, monoacetoxyscirpenol, diacetoxyscirpenol, T-2 triol, and T-2 tetraol) by liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) is presented. The development of the method and investigations on the matrix influence on the MS signal are described in particular. The matrix effect was thereby minimized by using an internal standard, a special mobile phase, and specific fragmentation parameters. The sample was extracted with acetonitrile/water (84:16, v/v), and the extract was cleaned up with a MycoSep 227 column. Quantification was based on the internal standard de-epoxy-deoxynivalenol. Calibration curves were linear between 16 and 1600 ng/g, and the limits of detection ranged from 0.18 to 5.0 ng/g. The developed method was applied for the determination of trichothecenes in 120 naturally contaminated wheat and oat samples.     

Simultaneous quantification of vitamin E and vitamin E metabolites in equine plasma and serum using LC-MS/MS

Vitamin E deficiencies can impact normal growth and development in humans and animals, and assessment of circulating levels of vitamin E and its metabolites may be an important endpoint for evaluation. Development of a sensitive method to detect and quantify low concentrations of vitamin E and metabolites in biological specimens allows for a proper diagnosis for patients and animals that are deficient. We developed a method to simultaneously extract, detect, and quantify the vitamin E compounds alpha-tocopherol (α-TP), gamma-tocopherol (γ-TP), alpha-tocotrienol (α-TT), and gamma-tocotrienol (γ-TT), and the corresponding metabolites formed after β-oxidation of α-TP and γ-TP, alpha-carboxymethylbutyl hydroxychroman (α-CMBHC) and alpha- or gamma-carboxyethyl hydroxychroman (α- or γ-CEHC), respectively, from equine plasma and serum. Quantification was achieved through liquid chromatography–tandem mass spectrometry. We applied a 96-well high-throughput format using a Phenomenex Phree plate to analyze plasma and serum. Compounds were separated by using a Waters ACQUITY UPLC BEH C18 column with a reverse-phase gradient. The limits of detection for the metabolites and vitamin E compounds were 8–330 pg/mL. To validate the method, intra-day and inter-day accuracy and precision were evaluated along with limits of detection and quantification. The method was then applied to determine concentrations of these analytes in plasma and serum of horses. Alpha-TP levels were 3–6 µg/mL of matrix; the metabolites were found at much lower levels, 0.2–1.0 ng/mL of matrix.     

Efficient cell reprogramming as a target for functional‐marker strategies? Towards new perspectives in applied plant‐nutrition research

The review aims at visualizing and strengthening approximation of current strategies in plant breeding, plant nutrition, and molecular biology. Innovations in new breeding strategies on quantitative traits are based on the development of functional DNA markers. This requires knowledge on robust physiological key reactions or parameters in view of the desired agronomic trait. To understand the significance of adaptive molecular‐physiological factors for the expression of agronomic traits in quantitative terms, systems analyses have to demonstrate the phenotypic effect of differential gene activities. The logistic to advance in applied systems biology is currently being strongly discussed. In the present contribution, identification of target cells, which are important for agronomic traits, is stressed as a key for future modeling and virtual experimentation. Integration of target cells in systems analysis should allow to link top‐down approaches, that start at the whole‐plant level, with bottom‐up approaches, that come from the molecular level. To illustrate the importance of adaptive cell reprogramming for agronomic traits, reprogramming of rhizodermic cells to trichoblasts is pointed out in its role for nutrient efficiency (NE). The nature of molecular factors, which may serve as functional markers in breeding, is discussed in view of future marker developments.     

Towards understanding the ecology and mechanisms of biocontrol of Clonostachys rosea IK726

Clonostachys rosea (syn. Gliocladium roseum) IK726 was originally selected as an effective biocontrol agent (BCA) against cereal seed borne diseases caused by Fusarium culmorum and Bipolaris sorokiniana. We have studied the efficacy of the antagonist against different pathogens in several crops and found that the antagonist also is able to control Alternaria radicina and A. dauci on carrot seeds and different cold-storage fungi in acorns. IK726 is also able to reduce…     

Levantilides A and B, 20-membered macrolides from a Micromonospora strain isolated from the Mediterranean deep sea sediment

Two new 20-membered macrolides, levantilide A and B, were isolated from the Micromonospora strain M71-A77. Strain M71-A77 was recovered from an Eastern Mediterranean deep-sea sediment sample and revealed to produce the levantilides under in situ salinity of 38.6 ‰. The chemical structures of the levantilides were elucidated on the basis of different one- and two- dimensional NMR experiments. Levantilide A exhibits a moderate antiproliferative activity against several tumor cell lines.     

Chondrosarcoma in a simmental cow: Clinical, ultrasonographic, radiographic and pathological findings

Clinical, ultrasonographic, radiographic and pathological findings of a chondrosarcoma diagnosed in an 8-year-old Simmental cow are reported. The primary site was located in the cartilage of the right scapula at the angulus cranialis. The tumour had already spread to the lungs at the time of diagnosis. This is the first report of a bovine scapula chondrosarcoma.     

Lack of glucuronidation products of trans‐resveratrol in plasma and urine of cats

Resveratrol has generated interest in cats due to reported health benefits. Cats have low activity of β‐glucuronidase, and we hypothesized they could not form two common resveratrol metabolites, resveratrol‐3‐O‐glucuronide and resveratrol‐4′‐O‐glucuronide. Resveratrol, 3 mg/cat/day, was given orally to intact male (n = 5) and female cats (n = 5) for 4 weeks. A control group (8 intact males) was used for comparison. Plasma and urine were collected weekly and analysed using high‐pressure liquid chromatography coupled with tandem mass spectrometry. Resveratrol and resveratrol‐3‐O‐sulphate, but no glucuronide metabolites, were detected in plasma and urine. Median (range 10–90th percentile) plasma resveratrol for control and treatment groups was 0.46 ng/ml (0.02–1.74 ng/ml) and 0.96 ng/ml (0.65–3.21 ng/ml). Median (range) plasma resveratrol‐3‐O‐sulphate for control and treatment groups was 6.32 ng/ml (2.55–10.29 ng/ml) and 11.45 ng/ml (1.47–53.29 ng/ml). Plasma resveratrol differed from control in week 4, while plasma resveratrol‐3‐O‐sulphate was different in all weeks (p < 0.05). Median (range) urine resveratrol for control and treatment groups was 0.28 ng/ml (0.05–1.59 ng/ml) and 19.98 ng/ml (8.44–87.54 ng/ml). Median (range) urine resveratrol‐3‐O‐sulphate for control and treatment groups was 26.71 ng/ml (10.50–75.58 ng/ml) and 108.69 ng/ml (11.83–231.05 ng/ml). All time points for urine resveratrol and resveratrol‐3‐O‐sulphate were significantly different from control (p < 0.05), except for weeks 1, 3 and 4 for resveratrol. The results support our hypothesis that cats are unlikely able to glucuronidate resveratrol, most likely due to a reduction in the activity of β‐glucuronidase.