Isolation and characterization of two actins of the Pacific white shrimp, <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis>

Two actin genes named actinT1 and actinT2 were isolated and sequenced from the Pacific white shrimp, Litopenaeus vannamei, by screening from a shrimp eyestalk cDNA library. L. vannamei actinT1 cDNA has a 1,128-bp open reading frame encoding for 376 amino acids while L. vannamei actinT2 cDNA has a 1,131-bp open reading frame coding for 377 amino acids. Alignment of the actinT1 and actinT2 cDNA sequences showed that these two actin genes share a sequence identity of 86% at amino acid residues. When compared with actins of several other invertebrate and vertebrate species, the nucleotide sequence of actinT1 is highly homologous (97–100%) with beta-actins, while actinT2 shares 86–95% identity with alpha-actins on the nucleotide level. Phylogenetic analysis and BLAST searches indicated that the ActinT1 protein is identical to crustacean beta-actins, while the ActinT2 protein is highly homologous to crustacean alpha-actins. Constitutive expression of the actinT1 and actinT2 genes were detected by RT-PCR in all adult shrimp organs, including brain, eye-stalk, gill, heart, hemolymph, hepatopancreas, muscle, swimming legs, and stomach, as well as in the shrimp zygote, nauplius, and mysis life stages. These data will facilitate attempts to clone and identify more shrimp genes and constitutive shrimp promoters.     

Enzyme-linked immunosorbent assay for bisphenol A: Assay optimization and its application for surface water analysis

A range of ELISAs in indirect and direct formats for the determination of bisphenol A (BPA) was developed. Bisphenol A carboxymethyl ether (BPA-CME), (BPA-CPE) and 4,4-bis(4-hydroxyphenyl)valeric acid were coupled with bovine serum albumin (BSA) (immunogens for the production of polyclonal antibodies), ovalbumin (OVA) and horseradish peroxidase. In general, the indirect assay was more sensitive and specific than the direct one. Using heterologous combinations of immunoreagents in the indirect assay allowed to increase assay sensitivity and specificity. The highest sensitivity was obtained for the antibodies produced against a conjugate of BPA-CPE with BSA and a conjugate of 4,4-bis(4-hydroxyphenyl)valeric acid with OVA. The detection limit of BPA in phosphate buffer was 0.03?ng?mL^?1. The assay developed was also the most specific towards BPA. Maximum cross-reactivity values did not exceed 11% for 4-cumylphenol, 5% for bisphenol E and 2% for bisphenol S. Finally, the developed assay was used to analyze surface water samples spiked with known amount of BPA. The assay showed good recovery values (85–109%) for surface water with mineralization level lower than 500?mg?L^?1.     

When good neighbors don't need fences: temporal landscape partitioning among baboon social groups

Intraspecific competition is a key factor shaping space-use strategies and movement decisions in many species, yet how and when neighbors utilize shared areas while exhibiting active avoidance of one another is largely unknown. Here, we investigated temporal landscape partitioning in a population of wild baboons (Papio cynocephalus). We used global positioning system (GPS) collars to synchronously record the hourly locations of five baboon social groups for ～900 days, and we used behavioral, demographic, and life history data to measure factors affecting use of overlap areas. Annual home ranges of neighboring groups overlapped substantially, as predicted (baboons are considered non-territorial), but home ranges overlapped less when space use was assessed over shorter time scales. Moreover, neighboring groups were in close spatial proximity to one another on fewer days than predicted by a null model, suggesting an avoidance-based spacing pattern. At all time scales examined (monthly, biweekly, and weekly), time spent in overlap areas was greater during time periods when groups fed on evenly dispersed, low-quality foods. The percent of fertile females in social groups was negatively correlated with time spent in overlap areas only during weekly time intervals. This suggests that broad temporal changes in ecological resources are a major predictor of how intensively overlap areas are used, and groups modify these ecologically driven spacing patterns at short time scales based on female reproductive status. Together, these findings offer insight into the economics of territoriality by highlighting the dynamics of spacing patterns at differing time scales.     

Selection for male choice based primarily on mate compatibility in the oldfield mouse, Peromyscus polionotus rhoadsi

Despite the consensus that mate choice acts as a mechanism for selection of secondary sexual traits, the evolutionary forces affecting mate preferences themselves remain controversial. In this study, we first demonstrated selection acting directly on the mate preferences of monogamous male oldfield mice, Peromyscus polionotus rhoadsi. One group of male oldfield mice were allowed to express a social preference between two potential mates, and were subsequently paired with either their preferred or rejected female. Among these pairs, those containing preferred females produced more offspring than did those containing rejected females. We next demonstrated that this fitness advantage depended primarily on compatibility between the members of a mated pair. A second group of male oldfield mice were not allowed the opportunity to express a social preference between potential mates. Rather, these males were paired with females that had been either preferred or rejected by males in the first group. Among these pairs, those containing preferred females did not produce more offspring than those containing rejected females. In other words, individual mate preferences had fitness consequences only for those males that expressed them, demonstrating that these preferences were based primarily on compatibility between mates.     

A framework to measure the wildness of managed large vertebrate populations

As landscapes continue to fall under human influence through habitat loss and fragmentation, fencing is increasingly being used to mitigate anthropogenic threats and enhance the commercial value of wildlife. Subsequent intensification of management potentially erodes wildness by disembodying populations from landscape-level processes, thereby disconnecting species from natural selection. Tools are needed to measure the degree to which populations of large vertebrate species in formally protected and privately owned wildlife areas are self-sustaining and free to adapt. We devised a framework to measure such wildness based on 6 attributes relating to the evolutionary and ecological dynamics of vertebrates (space, disease and parasite resistance, exposure to predation, exposure to limitations and fluctuations of food and water supply, and reproduction). For each attribute, we set empirical, species-specific thresholds between 5 wildness states based on quantifiable management interventions. We analysed data from 205 private wildlife properties with management objectives spanning ecotourism to consumptive utilization to test the framework on 6 herbivore species representing a range of conservation statuses and commercial values. Wildness scores among species differed significantly, and the proportion of populations identified as wild ranged from 12% to 84%, which indicates the tool detected site-scale differences both among populations of different species and populations of the same species under different management regimes. By quantifying wildness, this framework provides practitioners with standardized measurement units that link biodiversity with the sustainable use of wildlife. Applications include informing species management plans at local scales; standardizing the inclusion of managed populations in red-list assessments; and providing a platform for certification and regulation of wildlife-based economies. Applying this framework may help embed wildness as a normative value in policy and mitigate the shifting baseline of what it means to truly conserve a species.     

Cholinesterase activities as potential biomarkers: characterization in two freshwater snails, Potamopyrgus antipodarum (Mollusca, Hydrobiidae, Smith 1889) and Valvata piscinalis (Mollusca, Valvatidae, Müller 1774)

Anti-cholinesterase insecticides constitute a major portion of modern synthetic pesticides and the assessment of cholinesterase (ChE) inhibition is widely used as a specific biomarker for evaluating the exposure of non-target organisms to these pollutants. However, most studies on this biomarker were developed on vertebrates and among invertebrates, gastropod mollusks are rarely used. Gastropods are important members of aquatic habitats and therefore present a high ecological relevance for freshwater ecosystems. In this context, ChE activities were characterized in two freshwater gastropod mollusks, Potamopyrgus antipodarum and Valvata piscinalis, in order to ascertain their value as sentinel species. Firstly, characterization of ChE activities was performed using different substrates (acetylcholine iodide, butyrylcholine iodide and propionylcholine iodide) and specific inhibitors (eserine, iso-OMPA and BW284c51). Secondly, in vivo effect of a widely used organophosphate insecticide, chlorpyrifos, was tested on ChE activity in both species. Results suggested that P. antipodarum possesses two isoforms of cholinesterases, one isoform which properties are intermediate between an acetyl and a propionyl ChE, and one minor isoform which correspond to a butyryl ChE, while V. piscinalis seems to possess only one isoform which displays typical properties of an acetyl ChE. Chlorpyrifos induced no effect on V. piscinalis ChE. In contrast, P. antipodarum activity was significantly decreased by environmental realistic chlorpyrifos concentrations (2.86 and 14.2 nM) after seven days of contact. The present study suggests that P. antipodarum may be employed as a biological indicator for assessing pesticide contamination.     

Leaching of azoxystrobin and its degradation product R234886 from Danish agricultural field sites

The objective was to estimate leaching of the fungicide azoxystrobin (methyl (αE)-2-[[6-(2-cyanophenoxy)-4-pyrimidinyl]oxy]-α-(methoxymethylene)benzene-acetate) and one of its primary degradation products R234886 ([(E)-2-(2-[6-cyanophenoxy)-pyrimidin-4-yloxyl]-phenyl-3-methoxyacrylic acid], major fraction) at four agricultural research fields (one sandy and three loamy) in Denmark. Water was sampled from tile drains, suction cups and groundwater wells for a minimum period of two years after application of azoxystrobin. Neither azoxystrobin nor R234886 were detected at the sandy site, but did leach through loamy soils. While azoxystrobin was generally only detected during the first couple of months following application, R234886 leached for a longer period of time and at higher concentrations (up to 2.1 μg L⁻¹). Azoxystrobin is classified as very toxic to aquatic organisms and R234886 as very harmful. Our study shows that azoxystrobin and R234886 can leach through loamy soils for a long period of time following application of the pesticide and thereby pose a potential threat to vulnerable aquatic environments and drinking water resources. We thus recommend the inclusion of azoxystrobin and R234886 in pesticide monitoring programmes and further investigation of their long-term ecotoxicological effects.     

Towards a renewed research agenda in ecotoxicology

New concerns about biodiversity, ecosystem services and human health triggered several new regulations increasing the need for sound ecotoxicological risk assessment. The PEER network aims to share its view on the research issues that this challenges. PEER scientists call for an improved biologically relevant exposure assessment. They promote comprehensive effect assessment at several biological levels. Biological traits should be used for Environmental risk assessment (ERA) as promising tools to better understand relationships between structure and functioning of ecosystems. The use of modern high throughput methods could also enhance the amount of data for a better risk assessment. Improved models coping with multiple stressors or biological levels are necessary to answer for a more scientifically based risk assessment. Those methods must be embedded within life cycle analysis or economical models for efficient regulations. Joint research programmes involving humanities with ecological sciences should be developed for a sound risk management.