Rapid Biotic and Abiotic Transformation of Toxins produced by Ostreopsis. cf. ovata

The dinoflagellate Ostreopsis cf. ovata produces several families of toxic polyketides. Despite only a few field measurements of these phycotoxins in seawater and aerosols, they are believed to be responsible for dermatitis and the toxic inhalations reported during blooms of this species. Therefore, the stability of these compounds in seawater is essential to understanding the causes of these symptoms, however, this has never been assessed. In the current study, the optimization of a solid phase extraction (SPE) procedure was first performed to ensure the most efficient extraction of all phycotoxins known to be produced by this strain, including the recently described liguriatoxins. The SPE cartridge SDBL^® under non acidified conditions offered the best option. The stability of the ovatoxins and the liguriatoxins under biotic and abiotic stress was assessed by exposing the spent medium of a culture of Ostreopsis cf. ovata to its bacterial consortium and natural sunlight. A rapid biotic transformation was detected for both families of compounds. When exposed to bacteria, the half-lives of the ovatoxins were reached before 10 h and at 36 h, 97% of these toxins had been transformed. The half-lives of the liguriatoxins were 10 h under these conditions. Photolysis (abiotic degradation) of the ovatoxins (T_1/2 < 36 h) was faster than for the liguriatoxins (T_1/2 > 62 h). Although none of the catabolites of these phycotoxins were thoroughly identified, an untargeted metabolomics approach combined with molecular networking highlighted the presence of several compounds exhibiting structural similarities with the ovatoxins. Additional work should confirm the preliminary findings on these potential ovatoxins’ catabolites and their biological properties. The rapid transformation of O. cf. ovata’s phycotoxins introduces questions concerning their presence in seawater and their dispersion in the sea spray aerosols. The compounds involved in the toxic inhalations and dermatitis often experienced by beachgoers may stem from the catabolites of these toxins or even unrelated and as yet unidentified compounds.     

Multiscale comparison of spatial patterns using two-dimensional cross-spectral analysis: application to a semi-arid (gapped) landscape

Spectral analysis allows the characterization of temporal (1D) or spatial (2D) patterns in terms of their scale (frequency) distribution. Cross-spectral analysis can also be used to conduct independent correlation analyses at different scales between two variables, even in the presence of a complex superposition of structures, such as structures that are shifted, have different scales or have different levels of anisotropy. These well-grounded approaches have rarely been applied to two-dimensional ecological datasets. In this contribution, we illustrate the potential of the method. We start by providing a basic methodological introduction, and we clarify some technical points concerning the computation of two-dimensional coherency and phase spectra and associated confidence intervals. First, we illustrate the method using a simple theoretical model. Next, we present a real world application: the case of patterned (gapped) vegetation in SW Niger. In this example, we investigate the functional relationships between topography and the spatial distribution of two shrub species, Combretum micranthum G. Don. and Guiera senegalensis J.F. Gmel. We show that both the global vegetation pattern and the distribution of C. micranthum are independent at all analyzable scales (i.e., from 10 to 50 m) from possible relief-induced determinisms. Additionally, the two dominant shrub species form distinct patches, thus suggesting separate niches.     

Wnt gradient formation requires retromer function in Wnt-producing cells

Wnt proteins function as morphogens that can form long-range concentration gradients to pattern developing tissues. Here, we show that the retromer, a multiprotein complex involved in intracellular protein trafficking, is required for long-range signaling of the Caenorhabditis elegans Wnt ortholog EGL-20. The retromer functions in EGL-20-producing cells to allow the formation of an EGL-20 gradient along the anteroposterior axis. This function is evolutionarily conserved, because Wnt target gene expression is also impaired in the absence of the retromer complex in vertebrates. These results demonstrate that the ability of Wnt to regulate long-range patterning events is dependent on a critical and conserved function of the retromer complex within Wnt-producing cells.     

Exploration of intra- and inter-population genetic diversity in <Emphasis Type="Italic">Hedysarum coronarium</Emphasis> L. by AFLP markers

Amplified fragment length polymorphism (AFLP) markers were used to characterize the genetic diversity within and among natural populations and cultivars of Hedysarum coronarium. Twelve populations within Tunisia were evaluated with three AFLP primer combinations. A total of 207 reproducible bands was detected of which 178 (86%) were polymorphic. The great discriminative power of AFLP markers and their ability to represent genetic relationships among Hedysarum plants was demonstrated. Genetic diversity within and among populations was assessed through Principal Component Analysis (PCA) and cluster analysis by using the Neighbor-joining clustering algorithm. AFLP technology has provided evidence of a high degree of intra- and inter-population genetic diversity in H. coronarium. AFLP banding patterns provided molecular markers correlated with the plants’ geotropism. In addition, AFLP markers can differentiate wild accessions from cultivars. Moreover, geographical origins did not correspond to population clustering.     

Factors affecting population dynamics of Eurasian woodcocks wintering in France: assessing the efficiency of a hunting-free reserve

The Eurasian woodcock Scolopax rusticola is a migratory bird of major importance for hunting, which is susceptible to habitat loss and the stochastic effects of severe winter weather. Conservation issues mostly concerned regulation of hunting, but the efficiency of hunting-free reserves has never been investigated. We studied causes of mortality and survival probabilities of 98 radio-tagged woodcocks in a reserve with no hunting and in an adjoining hunting area in Brittany (France). Predation, mostly by mammalian predators on fields at night, was similar among adults and yearlings, while hunting mortality was more important in yearlings. Overall winter survival probabilities were 0.86 ± SE 0.07 in adults and 0.63 ± SE 0.07 in yearlings. Survival probabilities of both age classes increased as birds spent more time in the reserve. Equality of predation in both reserve and hunting areas suggests an additive mortality due to hunting over the winter. Population matrix models predicted that such low survival probabilities cannot sustain long term viable populations. These results call for caution in the harvesting of woodcock populations wintering in western France and could be a forewarning of a decline. The regulation of hunting by setting bag limits or reducing the length of shooting seasons, or the creation of reserves might be appropriate tools for the sustainable management of woodcock populations.     

An optimised protocol for molecular identification of Eimeria from chickens

Molecular approaches supporting identification of Eimeria parasites infecting chickens have been available for more than 20 years, although they have largely failed to replace traditional measures such as microscopy and pathology. Limitations of microscopy-led diagnostics, including a requirement for specialist parasitological expertise and low sample throughput, are yet to be outweighed by the difficulties associated with accessing genomic DNA from environmental Eimeria samples. A key step towards the use of Eimeria species-specific PCR as a sensitive and reproducible discriminatory tool for use in the field is the production of a standardised protocol that includes sample collection and DNA template preparation, as well as primer selection from the numerous PCR assays now published. Such a protocol will facilitate development of valuable epidemiological datasets which may be easily compared between studies and laboratories. The outcome of an optimisation process undertaken in laboratories in India and the UK is described here, identifying four steps. First, samples were collected into a 2% (w/v) potassium dichromate solution. Second, oocysts were enriched by flotation in saturated saline. Third, genomic DNA was extracted using a QIAamp DNA Stool mini kit protocol including a mechanical homogenisation step. Finally, nested PCR was carried out using previously published primers targeting the internal transcribed spacer region 1 (ITS-1). Alternative methods tested included sample processing in the presence of faecal material, DNA extraction using a traditional phenol/chloroform protocol, the use of SCAR multiplex PCR (one tube and two tube versions) and speciation using the morphometric tool COCCIMORPH for the first time with field samples.     

The impact of introduced predators, light-induced mortality of fledglings and poaching on the dynamics of the Cory’s shearwater (Calonectris diomedea) population from the Azores, northeastern subtropical Atlantic

Cory’s shearwater (Calonectris diomedea) is a procellariiform seabird which breeds in the Mediterranean and the north-eastern subtropical Atlantic, and which is considered “Vulnerable” in Europe due to recent declines at some localities. In the Azores archipelago (Atlantic), the introduction of mammalian predators by man has led to petrels being extirpated from the main islands, except for Cory’s shearwater. Currently, the Azorean population of Cory’s shearwaters represents 65% of the species’ world population. However, its dynamics remains unknown, although: (1) numbers might have declined by 43% between 1996 and 2001, (2) on the main islands, the young suffer mortality from introduced mammals, poachers, and urban lights upon fledging, and (3) at sea, the level of fishery mortality remains unknown.To fill this gap, we conducted a 7-year demographic survey on a mammal-free islet in the Azores to determine adult survival rate using capture-mark-recapture of the breeders and to estimate fecundity. We also assessed urban mortality using the data from the rescue campaigns annually conducted in the archipelago. Urban mortality concerned about 6% of fledglings, but its importance greatly varied among islands. When rescue campaigns occur, the rate might drop below 0.5%. Overall, our simulations concerning the next 100 years do not allow excluding a decrease in Cory’s shearwater numbers in the Azores without rescue campaigns (by 87% under the least favourable scenario). Rescue campaigns should only slow down the decline unless, simultaneously, lower competition for nests compared to Vila islet allows adults to breed almost every year on the main islands and juvenile survival exceeds a threshold value. Since adult survival rate was high (>0.93), an eventual decline of the Azorean population of Cory’s shearwaters would probably not result from fishery mortality of adults, but rather from poor fledgling productivity and perhaps also from low survival during the first year at sea.     

Development of a real-time PCR method to quantify the PGPR strain Azospirillum lipoferum CRT1 on maize seedlings

Azospirillum lipoferum CRT1 is a promising phytostimulatory PGPR for maize, whose effect on the plant is cell density-dependent. A nested PCR method is available for detection of the strain but does not allow quantification. The objective was to develop a real-time PCR method for quantification of A. lipoferum CRT1 in the rhizosphere of maize seedlings. Primers were designed based on a strain-specific RFLP marker, and their specificity was verified under qualitative and quantitative PCR conditions based on successful CRT1 amplification and absence of cross-reaction with genomic DNA from various rhizosphere strains. Real-time PCR conditions were then optimized using DNA from inoculated or non-inoculated maize rhizosphere samples. The detection limit was 60 fg DNA (corresponding to 19 cells) with pure cultures and 4 × 10⁴ CFU equivalents g⁻¹ lyophilized sample consisting of mixture of rhizosphere soil and roots. Inoculant quantification was effective down to 10⁴ CFU equivalents g⁻¹. Assessment of CRT1 rhizosphere levels in a field trial was in accordance with estimates from semi-quantitative PCR targeting another locus. This real-time PCR method, which is now available for direct rhizosphere monitoring of A. lipoferum CRT1 in greenhouse and field experiments, could be used as a reference for developing quantification tools for other Azospirillum inoculants.     

Low impact of present and historical landscape configuration on the genetics of fragmented Anthyllis vulneraria populations

Decreasing habitat fragment area and increasing isolation may cause loss of plant population genetic diversity and increased genetic differentiation between populations. We studied the relation between the historical and the present landscape configuration (i.e., patch area and patch connectivity), and the present management of calcareous grassland fragments on the one hand, and the within and between population genetic structure of 18 Anthyllis vulneraria populations on the other hand. Despite the long-time fragmentation history and the mainly selfing breeding system of the species, we detected very low genetic differentiation (Φ_st = 0.056) among habitat fragments and no significant isolation-by-distance relation. Average within fragment genetic diversity measured as molecular variance and expected heterozygosity, were relatively high (16.46 and 0.28, respectively), and weakly positively correlated with the current fragment area, most likely because larger fragments contained larger populations. We found no effects of the historical landscape configuration on the genetic diversity of the populations. Our data suggest that the consequences of habitat fragmentation for genetic differentiation and genetic diversity of A. vulneraria are relatively minor which is very likely due to the historical high levels of seed exchange among fragments through grazing and roaming livestock. This study provides indirect evidence that nature management by grazing not only positively affects habitat quality but that it might also mitigate the genetic consequences of habitat fragmentation. From the conservation point of view, this study illustrates the importance of grazing and of the regular transport of livestock between fragments to prevent the long-term effects of fragmentation on the genetic diversity of the populations studied.