Ultrastructure of spermatozoa of <Emphasis Type="Italic">Onthophagus taurus</Emphasis> (Coleoptera,Scarabaeidae) exhibits heritable variation

Sperm competition is thought to be an important selective pressure shaping sperm form and function. However, few studies have moved beyond gross examinations of sperm morphology. Sperm length is subject to sexual selection via sperm competition in the scarab beetle Onthophagus taurus. Here, the structure and ultrastructure of spermatozoa in this species were investigated using light and electron microscopy. Spermatozoa were found to be filiform, measuring about 1,200 mm in length. The sperm head consists of a three-layered acrosome and a nuclear region bearing the anterior extension of the centriole adjunct. Acrosome and nuclear regions are bilaterally symmetric, with their axes of symmetry being orthogonal to each other. Head and flagellar structures are connected by a well-developed centriole adjunct. The sperm heads are asymmetrically surrounded by accessory material and embedded into the cytoplasm of the spermatocyst cell. The accessory material is produced inside the spermatids and then transferred to the outside due to a new membrane formed around the sperm’s organelles. The old spermatid membrane separates the accessory material from the cyst cell. The flagellum contains a 9+9+2 axoneme, two accessory bodies, and two mitochondrial derivatives of unequal size. The major mitochondrial derivative is significantly larger than the minor one. The axoneme is arranged in a sinusoidal manner parallel along the major mitochondrial derivative. The spermatozoa show no progressive motility when released in buffer solution which is likely to be the result of the flagellar arrangement and the structure of the major mitochondrial derivative. The cross-sectional area of the minor and the major mitochondrial derivatives show different patterns of genetic variation. The data provide the first estimates of genetic variation in sperm ultrastructure for any species, and give evidence for the persistence of genetic variation in ultrastructure required for the rapid and divergent evolution that characterizes spermatozoa generally.     

Impact on sward composition and stock performance of grazing Molinia-dominant grassland

A comparative experiment was conducted to test the extent to which grazing of rank Molinia caerulea-dominated grassland increases botanical diversity and improves animal performance. The study was carried out on an area of under-grazed Molinia-dominant pasture that had not been stocked for 20 years. Three treatments were compared from 2001 until 2008: (1) no grazing, (2) grazing by Welsh Mountain sheep and (3) grazing by Welsh Black heifers. Patterns of change in sward surface composition across the grazing season were consistent with greater consumption of Molinia by the cattle, but there was little evidence of management regime influencing changes in sward composition year-on-year. There was a significant effect of year on animal performance (P < 0.001). Although the growth rates of the heifers improved over the course of the first three years of the experiment, this performance was not sustained. In contrast, any weight changes for the sheep were positive. The results highlight the slow pace of change within upland swards, and the need to develop site-specific grazing guidelines.     

Application of winery and distillery waste composts to a Jumilla (SE Spain) vineyard: Effects on the characteristics of a calcareous sandy-loam soil

The effects on different physico-chemical, chemical and biological soil properties of the incorporation of different organic materials into a calcareous vineyard soil were studied in an experiment during three growing seasons. The organic materials used were sheep manure and four composts derived from the treatment of winery and distillery wastes. A multivariate technique, factor analysis (FA), was also used for interpreting the data obtained from the samplings carried out throughout the three vine growing seasons. The application of the organic materials induced an increase in the activity of the soil microorganisms and in the soil macro and micronutrient contents, as well as a slow release of inorganic N. Through FA of the soil properties studied, three factors that explained 74.7% of the variability were established and the effects of the treatments after the whole experimental period were assessed.     

Anatomical localization and stereoisomeric composition of <Emphasis Type="Italic">Tribolium castaneum</Emphasis> aggregation pheromones

We report that the abdominal epidermis and associated tissues are the predominant sources of male-produced pheromones in the red flour beetle, Tribolium castaneum and, for the first time, describe the stereoisomeric composition of the natural blend of isomers of the aggregation pheromone 4,8-dimethyldecanal (DMD) in this important pest species. Quantitative analyses via gas chromatography–mass spectrometry showed that the average amount of DMD released daily by single feeding males of T. castaneum was 878 ± 72 ng (SE). Analysis of different body parts identified the abdominal epidermis as the major source of aggregation pheromone; the thorax was a minor source, while no DMD was detectable in the head. No internal organs or obvious male-specific glands were associated with pheromone deposition. Complete separation of all four stereoisomers of DMD was achieved following oxidation to the corresponding acid, derivatization with (1R, 2R)- and (1S, 2S)-2-(anthracene-2,3-dicarboximido)cyclohexanol to diastereomeric esters, and their separation on reversed-phase high-performance liquid chromatography at −54°C. Analysis of the hexane eluate from Porapak-Q-collected volatiles from feeding males revealed the presence of all four isomers (4R,8R)/(4R,8S)/(4S,8R)/(4S,8S) at a ratio of approximately 4:4:1:1. A walking orientation bioassay in a wind tunnel with various blends of the four synthetic isomers further indicated that the attractive potency of the reconstituted natural blend of 4:4:1:1 was equivalent to that of the natural pheromone and greater than that of the 1:1 blend of (4R,8R)/(4R,8S) used in commercial lures.     

Intra-cultivar variability of the soil-to-grain transfer of fallout 137Cs and 90Sr for winter wheat

Differences between the root uptake of fallout radionuclides by different cultivars ('inter-cultivar' variability) growing on the same field may be influenced not only by genetic differences of the cultivars, but also by the spatial variability of the soil-to-grain transfer within the cultivation area of each cultivar. This 'intra-cultivar' variability was investigated in 2001 and 2002 for 137Cs and 90Sr using three winter wheat cultivars with four replicates for each cultivar at three different sites in Bavaria, Germany. The intra-cultivar variability proved to be in the same range as the inter-cultivar variability which was determined earlier at the same sites for both radionuclides. An ANOVA of the 137Cs data set revealed that the variability of the 137Cs soil-to-grain transfer was caused by the soil and climate (year) at the field sites and the interaction of cultivar and field. A significant contribution of the factor 'cultivar' alone to the variability could not be detected. This may be due to the complex environmental conditions to which plants are exposed in field experiments. To find wheat cultivars with minimal uptake of fallout radionuclides it may be better to examine the molecular mechanisms of their root uptake in order to identify targets for breeding "safer" plants.     

Trace element levels in foetus-mother pairs of short-beaked common dolphins (Delphinus delphis) stranded along the French coasts

Tissues of foetus-mother pairs of common dolphins (Delphinus delphis) stranded along the French coasts (Bay of Biscay and English Channel) were analysed for their Cd, Cu, Hg, Se and Zn contents. In the kidneys, foetal Cd levels were extremely low, and strong relationships between Cu and Zn suggested the involvement of metallothioneins since early foetal life. The results also indicated a limited maternal transfer of Hg during pregnancy since levels in the tissues of foetuses were below 1 microg g(-1) w.wt. However, hepatic Hg levels in foetuses increased with body length, and were also proportionate to maternal hepatic, renal and muscular Hg levels. Lastly, affinities between Hg and Se in tissues would participate in Hg neutralisation in both mothers--through tiemannite granules--and fetuses--through reduced glutathione--counteracting the toxic effects linked to the particularly high quantities of methyl-Hg to which marine mammals are naturally exposed.     

Application of ICP-QMS for the determination of plutonium in environmental samples for safeguards purposes

Aiming to determine the plutonium amount as well as its isotopic composition, in particular, in swipe samples for safeguards purposes, an analytical method was developed with a plutonium separation step based on extraction chromatography using 2 cm TEVA columns and detection with quadrupole ICP-MS applying an ultra-sonic nebulizer coupled with membrane desolvation system. The method was successfully applied to New Brunswick plutonium certified reference materials as well as to Lawrence Livermore National Laboratory round robin samples, based on the round robin samples provided by the Institute for Reference Materials and Measurements (Belgium), as part of the Regular European Interlaboratory Measurement Evaluation Programme (REIMEP), campaign 16 (isotopic abundances of plutonium in plutonium nitrate samples), with a total plutonium amount between 1 and 0.25 ng per sample. After the introduction of an additional separation step, it was also possible to carry out precise and accurate total plutonium, (240)Pu/(239)Pu, (241)Pu/(239)Pu and (242)Pu/(239)Pu atom ratios determination in sediment sample showing its applicability to environmental samples in general, reaching a detection limit equivalent to 5 mBq(239)Pu kg(-1).     

The transfer of 239/240Pu to cow milk

In summer 1993 we measured the transfer of (239/240)Pu to milk from herbage from a pasture located 5 km from the Chernobyl nuclear power plant. In one treatment cows were allowed to graze freely on the pasture. In a second treatment, cows were fed herbage collected from the pasture in stalls. The milk transfer coefficient; F(m) did not vary significantly between treatments and the mean value of 7.5x10(-6)d l(-1) was higher than previously reported values. Despite many values of F(m) for Pu in the literature we identified few relevant original data sets. Transfer coefficient values for Pu are only appropriate when used in conjunction with a specified time period or an appropriate model which allows for the biological half-life. We recommend for screening purposes an F(m) value of 1x10(-5)d l(-1) for Pu, with an order of magnitude lower value being appropriate for cows which are only exposed for one grazing season.     

Generic approaches to transfer

A review of methods which have been used to describe and predict transfer of radionuclides in biota was undertaken. The intent was to identify approaches that might prove useful in extending predictive estimates to other organisms and environments. Empirical approaches, such as found in the use of transfer factors, were examined. Kinetic methodologies were also presented. Allometric functions, with their ability to make broad generalizations, were also discussed. Data from several earlier radioecological assessments were tested for their potential utility in developing allometric relationships, with the result implying that such an approach might prove useful.     

Presence, biotransformation and effects of sulfophenylcarboxylic acids in the benthic fish Solea senegalensis

The presence of linear alkylbenzene sulfonates (LAS) and their degradation intermediates, sulfophenylcarboxylic acids (SPCs), with concentrations up to 100 ppb has been found in surface waters taken from the estuary of the river Guadalete (Cádiz, SW of Spain). Higher concentrations were found at the sampling site located adjacent to the discharge outlet of a wastewater treatment plant (WWTP). The concentrations decreased downstream to a few ppb as a result of dilution, sorption, and degradation processes, which were taking place along the estuary. Once the presence of both xenobiotics was confirmed in the environment, an in vivo assay was conducted to study their biotransformation and effects in the benthic fish Solea senegalensis. A flow-through system was employed, consisting of an exposure phase (120 h) with environmental levels of the surfactant (200, 500 and 800 microg/L of 2?C(10)LAS), followed by a depuration stage (72 h). The generation of SPCs has been quantified during these phases in both water and fish, with LAS biotransformation shown in all cases. The antioxidant enzymes catalase (CAT), glutathione peroxidase (GPX), glutathione reductase (GR), the phase II enzyme glutathione S-transferase (GST), and the phase III acid and alkaline phosphatases (AcP, ALP) were also estimated and utilized as biomarkers.