Successful analysis of gut contents in fungal-feeding oribatid mites by combining body-surface washing and PCR

The feeding habits of soil mesofauna have been a mystery for decades, and depending on the methods used, different degrees of feeding specialisation have been observed. A new way to study the almost unknown feeding habits of soil mesofauna e.g. oribatid mites is to use PCR-based techniques. When applying PCR on these small organisms, the low amount of ingested DNA can cause problems. Even more important is to certify that the amplified DNA does not originate from body-surface contamination. The aim of this study was to analyse if washing of the body surface combined with PCR can be a successful approach when identifying the food ingested by fungivorous mites. The method was developed in a laboratory system where we used the oribatid mite Archegozetes longisetosus as a model organism due to its relatively short life cycle and ease of laboratory culturing. The results demonstrated that surface contamination is a serious problem. Both washing and dissection was needed to remove surface contamination on such small organisms. To get a reliable result the samples also had to contain at least five pooled guts, but preferably ten. This is the first step towards a successful use of PCR-based methods to study natural feeding habits of species most likely contaminated on the body surface. When modified for field conditions, the results obtained by this method have a high potential to answer many questions about the animals feeding habits, and their functional role in the soil.     

Carotenoid, chlorophyll, and chlorophyll-derived compounds in grapes and port wines

Carotenoids and chlorophyll-derived compounds in grapes and Port wines were investigated by HPLC-DAD and HPLC-DAD-MS (ESP+) analysis. A total of 13 carotenoid and chlorophyll-derived compounds are formally reported in grapes, 3 are identified for the first time, pheophytins a and b and (13Z)-beta-carotene, and 3 others remain unknown. In Port wines 19 compounds with carotenoid or chlorophyll-like structures are present, 8 still unidentified. The young wines showed higher total carotenoid content and chlorophyll-like compounds compared to aged Ports, with lutein and beta-carotene as major carotenoids. Among samples analyzed of monovarietal Vitis vinifera L. cultivar wines produced with the five most important Douro varieties, Tinta Roriz contained the highest levels of carotenoids and Touriga Franca the lowest. The forced-aging study indicated that lutein was more sensitive to temperature than beta-carotene. Additionally, aged wines showed higher ratios of beta-carotene/lutein concentrations compared to new Ports. Rates of degradation of chlorophyll derivative compounds were higher than those for carotene and lutein.     

Purification and partial characterization of lipoxygenase from desert truffle (Terfezia claveryi Chatin) ascocarps

A lipoxygenase from Terfezia claveryi Chatin ascocarp, a mycorrhizal hypogeous fungus, is described for the first time. The higher proportion of PUFA in T. claveryi ascocarps makes lipid rancidity the main factor limiting its storage life. Thus, the studies on LOX from T. claveryi are important because this enzyme, among other roles, may be involved in an alteration of lipids leading to consumer rejection. The enzyme has been purified to apparent homogeneity by phase partitioning in the presence of Triton X-114, followed by two steps of cation-exchange chromatography. The purified T. claveryi LOX preparation consisted of a single major band with an apparent molecular mass of 66 kDa after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzymic activity exhibited a strong specificity toward linoleic and linolenic acids as substrates, while only 32% activity was observed using gamma-linolenic acid. The pH optimum of this enzyme was pH 7.0. When the enzyme reacted with linoleic acid, it produced a single peak, which comigrated with standard 13-hydroperoxy-octadecadienoic acid; 13-hydroperoxy-octadecatrienoic acid was produced during the reaction with linolenic acid.     

Interaction of contaminated sediment from a salt marsh with estuarine water: evaluation by leaching and ecotoxicity assays and salts from leachate evaporation

Purpose

Wastes from a former Portuguese steel plant were deposited between 1961 and 2001 on the riverbank of a tributary of the Tagus River creating a landfill connected to the river, posing a potential contamination risk to the Tagus estuary ecosystem. This study aims to assess the transfer of chemical elements from contaminated sediments to the estuarine water from cycles of sediment leaching so as to evaluate the ecotoxicity of the leachates, and to analyze the solid phases crystallized from those leachates.

Materials and methods

Landfill sediment and estuarine water samples were collected during low tide. Sediment samples were analyzed for pH, electric conductivity (EC), C_org, NPK, and iron oxides. Leaching assays (four replicates) were done using estuarine water (200 cm³/replicate) and 1.5 kg of sediment per reactor. Each reactor was submitted to four leaching processes (0, 28, 49, and 77 days). The sediment was kept moist between leaching processes. Sediment (total (acid digestion) and available fraction (diluted organic acid extraction-Rhizo)) elemental concentrations were determined by inductively coupled plasma–instrumental neutron activation analysis (ICP/INAA). Leachates, and estuarine and sediment pore waters were analyzed for metals/metalloids by ICP/mass spectrometry (MS) and carbonates/sulfate/chloride by standard methodologies. Ecotoxicity assays were performed in leachates and estuarine and pore waters using Artemia franciscana and Brachionus plicatillis. Aliquots of the leachates were evaporated to complete dryness (23–25 °C) and crystals analyzed by X-ray powder diffraction (XRD).

Results and discussion

Sediment with pH?=?8 and high EC and C_org was contaminated with As, Cd, Cr, Cu, Pb, and Zn. The element concentrations in the available fraction of the sediment were low compared to the sediment total concentrations (<1 % for Rhizo extraction). The concentrations of potentially hazardous elements in the estuarine water were relatively low, except for Cd. Concentrations of hazardous elements in the leachates were very low. Calcium, K, Mg, Na, and chloride concentrations were high but did not vary significantly among the four leaching experiments. Total concentrations of carbonate were much higher in leachates than in estuarine water. Both estuarine water and leachates showed negligible toxicity. Crystals identified in the solids obtained from the leachates by evaporation were halite, anhydrite, epsomite, dolomite, and polyhalite.

Conclusions

The sediment showed the capacity to retain the majority of the potentially hazardous chemical elements. Remobilization of chemical elements from sediment by leaching was essentially negligible. The variation of total concentrations of Ca, carbonate, and sulfate in leachates indicates that the sediment contained reactive sulfides. Due to its composition, the sediment seems to be a dynamic system of pollution control, which should not be disturbed.

     

Novel application of square-wave adsorptive-stripping voltammetry for the determination of xanthohumol in spent hops

This paper reports the development of a novel electrochemical assay for xanthohumol (XN) by square-wave adsorptive-stripping voltammetry (SWAdSV) with a hanging mercury drop electrode. The method showed good repeatability (CV < 2%) and linearity (between 10 and 250 μg L(-1)), as well as suitable limits of detection (2.6 μg L(-1)) and quantification (8.8 μg L(-1)). The method was applied for the quantification of this compound in spent hops, and the results obtained were compared with the HPLC-UV method. XN contents determined by the SWAdSV method were 16 ± 1 and 100 ± 4 μg L(-1) for aqueous and methanolic extracts, respectively. The developed new methodology considerably reduces the analysis time, approximately from 25 min (HPLC-UV method) to 7 min, enabling a high sample throughput. In addition, the detection and quantification limits were approximately 5-fold lower than those obtained with the chromatographic method.     

Long-term effect of toltrazuril on growth performances of dairy heifers and beef calves exposed to natural Eimeria zuernii and Eimeria bovis infections

The long-term effects of a toltrazuril treatment against natural Eimeria bovis and/or Eimeria zuernii infections were investigated in comparison with diclazuril and untreated controls on two dairy (Italian Friesian breed) and two beef (Chianina breed) farms. At each trial site, 30 calves were allocated into three groups of 10 calves each: T (treated with toltrazuril), D (treated with diclazuril) and C (left untreated). For 40 weeks post-treatment, the calves were weighed and examined clinically and parasitologically. The oocyst counts as well as the number of scour days were significantly lower in the T group than in the D and C groups. Final bodyweights and body condition scores of the T group exceeded those of groups C and D. The results confirmed that toltrazuril was highly efficacious, safe and provided productive benefits in dairy and beef calves.     

Monitoringergebnisse der Schnellk?fergattung Agriotes (Elateridae, Coleoptera) in den Jahren 2008 bis 2010 in Rheinland-Pfalz

During the years 2008 to 2010 beetles of the genus Agriotes were monitored by means of pheromone traps in Rhineland-Palatinate. Each trap contained the female sexual pheromone of either Agriotes obscurus, A. sordidus, A. lineatus, A. sputator, or A. ustulatus. Beetles of these species could be caught, additionally also specimens of the species A. gallicus, A. acuminatus, and A. pilosellus. Specimens of A. brevis and A. proximus could not be caught, not even in traps with pheromones according to their species. The high frequency of A. sputator in the Upper Rhine Valley and mild river valleys was surprising. This might be an explanation for the increasing damages especially in potatoes during the past seven years. Also the species A. ustulatus occurred mainly in the Rhine Valley. The species A. obscurus was found predominantly in higher altitudes, A. lineatus and A. sputator appeared in the whole area, with varying frequencies. In all, we could determine the composition of Agriotes-species in 27 fields. In two fields traps were exposed in two subsequent years, in one field in three years. The changes in the populations are shown. The beetles collected in some fields were compared with the according wireworms.     

A simple and sensitive liquid chromatography-mass spectrometry confirmatory method for analyzing sulfonamide antibacterials in milk and egg

A simple and specific method able to identify and quantify traces of 14 sulfonamide antibacterials (SAs) in milk and eggs is presented. This method uses a single solid-phase extraction (SPE) cartridge for simultaneous extraction and purification of SAs in the above matrices. Milk and egg samples are passed through a Carbograph 4 sorption cartridge. After analyte desorption, an aliquot of the final extract is injected into a liquid chromatography-mass spectrometry (LC-MS) instrument equipped with an electrospray ion source (ESI) and a single quadrupole. MS data acquisition is performed in the positive-ion mode and by a time-scheduled multiple-ion selected ion monitoring program. Compared to two published methods, the present protocol extracted larger amounts of SAs from both milk and egg and decreased the analysis time by a factor of 3 with milk samples and by a factor of 2 with egg samples. Recovery of SAs in milk at the 5 ppb level ranged between 76 and 112% with relative standard deviations (RSDs) of 相似文献