Geometric morphometrics as a tool for improving the comparative study of behavioural postures

Describing postures has always been a central concern when studying behaviour. However, attempts to compare postures objectively at phylogenetical, populational, inter- or intra-individual levels generally either rely upon a few key elements or remain highly subjective. Here, we propose a novel approach, based on well-established geometric morphometrics, to describe and to analyse postures globally (i.e. considering the animal’s body posture in its entirety rather than focusing only on a few salient elements, such as head or tail position). Geometric morphometrics is concerned with describing and comparing variation and changes in the form (size and shape) of organisms using the coordinates of a series of homologous landmarks (i.e. positioned in relation to skeletal or muscular cues that are the same for different species for every variety of form and function and that have derived from a common ancestor, i.e. they have a common evolutionary ancestry, e.g. neck, wings, flipper/hand). We applied this approach to horses, using global postures (1) to characterise behaviours that correspond to different arousal levels, (2) to test potential impact of environmental changes on postures. Our application of geometric morphometrics to horse postures showed that this method can be used to characterise behavioural categories, to evaluate the impact of environmental factors (here human actions) and to compare individuals and groups. Beyond its application to horses, this promising approach could be applied to all questions involving the analysis of postures (evolution of displays, expression of emotions, stress and welfare, behavioural repertoires…) and could lead to a whole new line of research.     

On the significance of adult play: what does social play tell us about adult horse welfare?

Play remains a mystery and adult play even more so. More typical of young stages in healthy individuals, it occurs rarely at adult stages but then more often in captive/domestic animals, which can imply spatial, social and/or feeding deprivations or restrictions that are challenging to welfare, than in animals living in natural conditions. Here, we tested the hypothesis that adult play may reflect altered welfare states and chronic stress in horses, in which, as in several species, play rarely occurs at adult stages in natural conditions. We observed the behaviour (in particular, social play) of riding school horses during occasional outings in a paddock and measured several stress indicators when these horses were in their individual home boxes. Our results revealed that (1) the number of horses and rates of adult play appeared very high compared to field report data and (2) most stress indicators measured differed between ‘players’ and ‘non-players’, revealing that most ‘playful’ animals were suffering from more chronic stress than ‘non-playful’ horses. Frequency of play behaviour correlated with a score of chronic stress. This first discovery of a relationship between adult play and altered welfare opens new lines of research that certainly deserves comparative studies in a variety of species.     

Genetic differentiation among matrilines in social groups of rhesus monkeys

Summary In rhesus monkey populations, animals related by descent to some female comprise a matriline or genealogy. Data on blood protein polymorphisms in the Cayo Santiago rhesus colony indicate that allele frequency variations among matrilines in social groups are large. These variations occur despite high levels of outbreeding. Computer simulation analyses indicate that pedigree or linear effect account for much of the observed genetic differentiation among genealogies. A sampling with correlation model in which genealogy sizes and average kinship levels are parameters predicts among matriline genetic differentiation. This study indicates that substantial genetic substructure is present within rhesus social groups. Our analyses also predict that large variances in allele frequencies should be common among social or trait groups based on kinship relationships.     

Toxic equivalency factors for PAH and their applicability in shellfish pollution monitoring studies

Fish and shellfish are exposed to a wide range of polycyclic aromatic hydrocarbons (PAH) following oil spills at sea, and can become contaminated as a result. Finfish have a more effective mixed-function oxidase enzyme system than shellfish, and are therefore able to metabolise and excrete PAH more effectively than the invertebrates. Thus, contamination by high-molecular weight PAH, including those with carcinogenic potential and so of concern with regard to human consumers, is therefore usually observed in shellfish, and particularly in bivalve molluscs. Oil spills are not the sole source of PAH, however, as parent compounds are also generated by a wide range of combustion processes. In this paper, consideration is given to monitoring data gathered following recent oil spills (both of crude oil and diesel fuel), alongside data from other studies. These include studies conducted around a former gasworks site and downstream of an aluminium smelter in the UK, and from mussel monitoring studies undertaken in the UK and the USA (including the Exxon Valdez oil spill and the National Status and Trends programme), and in other countries in Europe. For comparative purposes the PAH concentrations are summed and also expressed as benzo[a]pyrene equivalents, their relative concentrations being weighted in relation to the carcinogenic potential of individual PAH compounds using toxic equivalency factors (TEF). Our aim was to assess the utility of this approach in fishery resource monitoring and control following oil spills. Certainly this approach seems useful from the data assessed in this study. and the relative ranking of the various studies seems to reflect the relative degree of concern for human consumers due to the differing contamination sources. As a simple tool for control purposes it is equally applicable to PAH derived from oil spills, and from industrial and combustion sources.     

Rate of pesticide volatilization from soil: an experimental approach with a wind tunnel system applied to trifluralin

Pesticide volatilization to the atmosphere may be a major pathway of dissipation closely linked with environmental, physico-chemical and technical factors. Understanding the volatilization process requires systems that make it possible to control some of these factors. Wind tunnels meet to these criteria. The volatilization flux is determined from a mass balance, using the difference in atmospheric pesticide concentration between the entrance and the exit of the tunnel and the airflow rate. An experiment was carried out in June 2000 to study the repeatability of this technique. Volatilization of trifluralin was measured in three wind tunnels for 8 days with a sampling period varying between 3 h and 2 days. Pesticide concentration was determined by trapping by XAD-2 resin in a two-stage cartridge, solvent extraction and analysis by gas chromatography. Cumulated losses through volatilization reached 30% of the measured application dose after 8 days, with a variability of less than 20% between the three tunnels. Approximately 20% remained in the topsoil (0–2 cm), with a variability of 14% between the three tunnels. The decrease in the volatilization flux over time is coherent with the expected theoretical evolution for a volatile pesticide such as trifluralin and with previous experimental works.     

Socio-environmental implications of process-based restoration strategies in large rivers: should we remove novel ecosystems along the Rhône (France)?

River restoration efforts require interdisciplinary approaches involving fluvial geomorphology, hydraulic engineering, ecology, sedimentology, chemistry, social geography, and sociology. We investigated the functioning of artificial structures called “Casiers Girardon” (groyne fields) in the Rhône River. We assessed potential benefits and risks linked to removing the Rhône groyne fields in a restoration context, with particular focus on the potential for increased bank erosion. Hydraulic, morphological, chemical, ecological, and social issues resulting from dismantlement were studied for terrestrialized and aquatic Casiers Girardon. Only 10% of Casiers Girardon have maintained their aquatic features, whereas most of the Casiers are terrestrialized. Our results help to confirm the effectiveness of restoration actions; however, they also indicate uncertainties and additional knowledge needs, especially in regard to potential incompatibilities between Casier restoration and conservation. Then, an interdisciplinary conceptual model was developed to identify interventions to be considered in Casiers Girardon, according to their terrestrialization rate and physiochemical characteristics (connectivity, amount of gravel vs. fine sediment, contamination level). This model synthetizes scientific results and expert judgment and provides management recommendations based on ecological and sociological expectations about the restoration of Casiers Girardon. The model highlights high heterogeneity in functioning and ecological potential between terrestrialized and aquatic Casiers. Dismantling of terrestrialized Casiers has strong potential to provide multiple benefits, whereas aquatic Casiers could be maintained as valuable backwaters. The managing guidelines for the Casiers Girardon of the Rhône River should be adapted according to local conditions, as well as expected benefits and needs, and conducted in co-ordination with all actors involved in and affected by the restoration.     

Adaptation of the Daphnia sp. acute toxicity test: miniaturization and prolongation for the testing of nanomaterials

Manufacturing of nanomaterials (NMs) is often complex and expensive, and their environmental risks are poorly understood or even unknown. An economization of testing NMs is therefore desirable, which can be achieved by miniaturizing test systems. However, the downsizing of test vessels and volumes can enlarge the surface/volume ratio (SVR) which in turn can affect the bioavailable concentration of adsorbing substances like NMs. The present study focused on the miniaturization of the acute toxicity test with Daphnia magna. The adaptations were verified with three reference substances, the non-adsorbing potassium dichromate (K₂Cr₂O₇) and as potentially highly-adsorbing substances silver nanoparticles (AgNPs) and silver nitrate (AgNO₃). The miniaturized test was conducted in 24-well microtiter plates (MT) and simultaneously compared to the OECD standard test (ST). Furthermore, the test duration was prolonged from 48 to 96 h since NMs tend to show effects only after extended exposure. The toxicity of K₂Cr₂O₇ and AgNPs continued to increase within the prolonged test span. The test comparisons with K₂Cr₂O₇ did not reveal any significant differences between ST and MT. AgNO₃ toxicity was significantly decreased in MT compared to ST due to the enlarged SVR. The toxicity of AgNPs in MT after 24 h was equal to ST. Contrary to our expectations an exposure longer than 24 h resulted in an increase of AgNP toxicity in MT, possibly due to enhanced dissolution of silver. Microtiter plates are appropriate alternative test vessels for the Daphnia sp. acute toxicity test; thus, its miniaturization is feasible. The enlarged SVR has to be taken into account since it can affect the toxicity of potentially adsorbing substances. Furthermore, the standard test duration of 48 h might underestimate the toxicity of many substances, especially of NMs.     

The meta-hydroxylation step in the phenylpropanoid pathway: a new level of complexity in the pathway and its regulation

Together in silico and genetic mining approaches have recently designated the CYP98 family of plant cytochromes P450 as the family of enzymes that catalyzes the meta-hydroxylation step in the phenylpropanoid pathway. This meta-hydroxylation is not catalyzed on the free p-coumaric acid as anticipated, but on its conjugates with shikimic, quinic, or phenyllactic acids. While all CYP98s have in common phenol meta-hydroxylase activity, p-coumaroylshikimate remains their preferred substrate. High expression of CYP98s is detected in lignifying tissues in stems, roots, and siliques. The CYP98A3 gene disruption in Arabidopsis thaliana leads to a drastic inhibition of lignin synthesis, cell growth, and plant development. The meta-hydroxylation of phenolic precursors is thus essential for higher plant development. Isolation of coding sequences belonging to the CYP98 family from basil, wheat, and extensive functional analysis of the recombinant enzymes, together with CYP98s from other plant taxa, helps shedding some light on mechanisms of P450s evolution. Most importantly, the occurrence of the meta-hydroxylation on esters of shikimic or quinic acids introduces a new biochemical regulation mechanism in the phenylpropanoid pathway.