A reliable method for sexing unincubated bird eggs for studying primary sex ratio

In birds, offspring sex ratio manipulation by mothers is now well established with potentially important consequences for evolution and animal breeding. In most studies on primary sex ratio of birds, eggs are sexed after incubation by the use of PCR methods targeted to the sex-linked CHD1 genes. Sexing of unincubated eggs would be preferred, but as fertile and infertile blastodiscs cannot be distinguished macroscopically, errors could arise from PCR amplifications of parental DNA associated with the vitelline membrane of infertile eggs. In this study, we stained blastodiscs without the vitelline membrane with Hoechst 33342. This allowed unequivocal distinction between fertile and infertile blastodiscs. Fertile blastodiscs contained thousands of fluorescent nuclei, whereas no nuclei were seen in infertile eggs. In addition, after nucleic acid analysis, fertile blastodiscs yielded much stronger chromosomal DNA and CHD1-targeted PCR bands on agarose gels compared with infertile blastodiscs. These findings indicate that fertile blastodiscs contain much more embryonic DNA than parental DNA, allowing reliable sexing of the fertile eggs. The differences between fertile and infertile blastodiscs in chromosomal DNA and CHD1 PCR banding intensities alone could also be used to distinguish fertile from infertile eggs without using Hoechst staining. We conclude that identifying fertile blastodiscs either by Hoechst staining or by analyzing the yield of chromosomal DNA and CHD1-PCR products, combined with CHD1-targeted PCR amplification, presents an easy and reliable method to sex unincubated eggs.     

Inter-population variation in the behaviour of adult and juvenile Red-footed Boobies Sula sula

Early life is a critical phase of the life cycle of animals and is attracting increased attention because little information is available on the behaviour of young individuals during this period. Behaviour during early life is probably influenced by the environmental conditions encountered by young animals, but data on intraspecific variation between breeding sites during this crucial period of life are limited. Here we study variability in the foraging behaviour of juveniles and adults in three colonies of a pantropical seabird, the Red-footed Booby Sula sula. Both adults and juveniles were measured and fitted with GPS loggers in three remote islands: Genovesa (Galapagos, Eastern Pacific Ocean), Europa (Western Indian Ocean) and Surprise (New Caledonia, Western Pacific Ocean). Foraging behaviour was compared between age-classes, sex and colonies by examining trip characteristics, different behaviours at sea, potential associations between individuals and morphological characteristics. Compared with adults, juveniles conducted shorter trips that were restricted to around the colony, especially on Genovesa (max. range: 203.4 ± 125.1 km and 3.6 ± 3.1 km, respectively). Juveniles appeared more constrained by poor flight skills and experience rather than by their morphology. Adults travelled 45% of the time during at-sea trips, whereas juveniles spent a a lower proportion of time travelling but foraged more often using an ‘area-restricted search’ behaviour, potentially training to catch prey. Associations between juveniles were commonly detected in the three colonies and occurred mostly during foraging, suggesting that social learning is an important strategy. Variability of morphometric measurements in both adults and juveniles was high between sites, with larger birds found on Genovesa. These results suggest that adaptations to local environmental conditions are already visible in their early life. Future studies should continue to investigate the behavioural flexibility of juvenile birds to better understand the effect of local environmental conditions during this critical stage of life.     

Food dependent color patterns inThamnocephalus platyurus Packard (Branchiopoda: Anostraca); a laboratory study

Coloration of phyllopods varies from place to place and from one life stage to another. It ranges from translucent or whitish through gray, blue, green, orange, and reddish. Here, we present experimental evidence for a food- dependent color pattern inThamnocephalus platyurus Packard. The presence or absence of the synthetic pigment trans — — carotene in a baker's yeast diet was the controlling factor. All the 24 old larvae used in the experiment were whitish in color. From day 6 until the end the experiment (day 11), 100% of the shrimps under a diet with synthetic trans — — carotene (treatment 1) exhibited a characteristic color pattern which consisted of an orange color in the cercopods, and in all theracopods; the rest of the body exhibited no particular color. In comparison, 100% of the shrimps under a diet without synthetic trans — — carotene (treatment 2) were whitish throughout the body. In females from treatment 1, the ovaries and oocytes were green-bluish, while in females from treatment 2 the ovaries and oocytes were whitish. No significant differences in survival and growth were found, except that at day 9, there was a significant difference in growth, the females with the synthetic trans — — carotene group growing faster.     

Seasonal incidence of Ixodes ricinus ticks (Acari: ixodidae) on rodents in western France

Data collected from a longitudinal survey carried out over 2 years on four farms in western France were used to assess the incidence and infestation of Ixodes ricinus on rodents. Once a month, on each farm, 25 Sherman live traps were set in hedges bordering selected pastures. A total of 799 micromammals were examined, including Apodemus sylvaticus, Clethrionomys glareolus, Microtus agrestis, Microtus arvalis, and Crocidura spp. Larvae and nymphs of I. ricinus were found. Small numbers of Ixodes (Exopalpiger) trianguliceps were also recovered from each farm. The mean infestation rate of the I. ricinus larvae (1.6–5.9) among all animals examined varied between farms Most animals were infested by only a single tick, but one M. agrestis harboured 43 I. ricinus larvae. Larvae or nymphs were found throughout the year, with peaks from March to October.     

Structural basis of oligomannose recognition by the Pterocarpus angolensis seed lectin

The crystal structure of a Man/Glc-specific lectin from the seeds of the bloodwood tree (Pterocarpus angolensis), a leguminous plant from central Africa, has been determined in complex with mannose and five manno-oligosaccharides. The lectin contains a classical mannose-specificity loop, but its metal-binding loop resembles that of lectins of unrelated specificity from Ulex europaeus and Maackia amurensis. As a consequence, the interactions with mannose in the primary binding site are conserved, but details of carbohydrate-binding outside the primary binding site differ from those seen in the equivalent carbohydrate complexes of concanavalin A. These observations explain the differences in their respective fine specificity profiles for oligomannoses. While Man(alpha1-3)Man and Man(alpha1-3)[Man(alpha1-6)]Man bind to PAL in low-energy conformations identical with that of ConA, Man(alpha1-6)Man is required to adopt a different conformation. Man(alpha1-2)Man can bind only in a single binding mode, in sharp contrast to ConA, which creates a higher affinity for this disaccharide by allowing two binding modes.     

Partial modelling of the perennial crop cycle misleads LCA results in two contrasted case studies

Purpose

As highlighted in recent reviews, there is a need to harmonise the way life cycle assessment (LCA) of perennial crops is conducted. In most published LCA on perennial crops, the modelling of the agricultural production is based on data sets for just one productive year. This may be misleading since performance and impacts of the system may greatly vary year by year. The purposes of this study are to analyse how partial modelling of the perennial cycle through non-holistic data collection may affect LCA results and to make recommendations.

Methods

Three modelling choices for the perennial crop cycle were tested in parallel in two contrasted LCA case studies: oil palm fruits from Indonesia, and small citrus from Morocco. Modelling choices tested were as follows: (i) a chronological modelling over the complete crop cycle of orchards, (ii) a 3-year average from the productive phase, and (iii) various single years from the productive phase. In both case studies, the system boundary was a cradle-to-farm gate with a functional unit of 1 kg fresh fruits. LCA midpoint impacts were calculated with ReCiPe 2008 in Simapro©V.7. We first analysed how inputs, yields and potential impacts varied over time. We then analysed process contributions in the baseline model, i.e. the chronological modelling, and finally compared LCA results for the various perennial modelling choices.

Results and discussion

Agricultural practices, yields and impacts varied over the years especially during the first 3–9 years depending on the case study. In both case studies, the modelling choices to account or not for the whole perennial cycle drastically influenced LCA results. The differences could be explained by the inclusion or not of the yearly variability and the accounting or not of the immature phase, which contributed to 7–40 or 6.5–29 % of all impact categories for oil palm fruit and citrus, respectively.

Conclusions

The chosen approach to model the perennial cycle influenced the final LCA results for two contrasted case studies and deserved specific attention. Although data availability may remain the limiting factor in most cases, assumptions can be made to interpolate or extrapolate some data sets or to consolidate data sets from chronosequences (i.e. modular modelling). In all cases, we suggest that the approach chosen to model the perennial cycle and the representativeness of associated collected data should be made transparent and discussed. Further research work is needed to improve the understanding and modelling of perennial crop functioning and LCA assessment.

     

Demographic Responses to Oxidative Stress and Inflammation in the Wandering Albatross (Diomedea exulans)

One of the major challenges in ecological research is the elucidation of physiological mechanisms that underlie the demographic traits of wild animals. We have assessed whether a marker of plasma oxidative stress (TBARS) and plasma haptoglobin (protein of the acute inflammatory phase response) measured at time t predict five demographic parameters (survival rate, return rate to the breeding colony, breeding probability, hatching and fledging success) in sexually mature wandering albatrosses over the next four years (Diomedea exulans) using a five-year individual-based dataset. Non-breeder males, but not females, having higher TBARS at time t had reduced future breeding probabilities; haptoglobin was not related to breeding probability. Neither TBARS nor haptoglobin predicted future hatching or fledging success. Haptoglobin had a marginally positive effect on female survival rate, while TBARS had a marginally negative effect on return rate. Our findings do not support the role for oxidative stress as a constraint of future reproductive success in the albatross. However, our data point to a potential mechanism underlying some aspects of reproductive senescence and survival. Our results also highlight that the study of the consequences of oxidative stress should consider the life-cycle stage of an individual and its reproductive history.     

Purification and molecular cloning of a new galactose-specific lectin from <Emphasis Type="Italic">Bauhinia variegata</Emphasis> seeds

A new galactose-specific lectin was purified from seeds of a Caesalpinoideae plant, Bauhinia variegata, by affinity chromatography on lactose-agarose. Protein extracts haemagglutinated rabbit and human erythrocytes (native and treated with proteolytic enzymes), showing preference for rabbit blood treated with papain and trypsin. Among various carbohydrates tested, the lectin was best inhibited by D-galactose and its derivatives, especially lactose. SDS-PAGE showed that the lectin, named BVL, has a pattern similar to other lectins isolated from the same genus, Bauhinia purpurea agglutinin (BPA). The molecular mass of BVL subunit is 32 871 Da, determined by MALDI-TOF spectrometry. DNA extracted from B. variegata young leaves and primers designed according to the B. purpurea lectin were used to generate specific fragments which were cloned and sequenced, revealing two distinct isoforms. The bvl gene sequence comprised an open reading frame of 876 base pairs which encodes a protein of 291 amino acids. The protein carried a putative signal peptide. The mature protein was predicted to have 263 amino acid residues and 28 963 Da in size.     

Topological classification of RNA structures

We present a novel topological classification of RNA secondary structures with pseudoknots. It is based on the topological genus of the circular diagram associated to the RNA base-pair structure. The genus is a positive integer number whose value quantifies the topological complexity of the folded RNA structure. In such a representation, planar diagrams correspond to pure RNA secondary structures and have zero genus, whereas non-planar diagrams correspond to pseudoknotted structures and have higher genus. The topological genus allows for the definition of topological folding motifs, similar in spirit to those introduced and commonly used in protein folding. We analyze real RNA structures from the databases Worldwide Protein Data Bank and Pseudobase and classify them according to their topological genus. For simplicity, we limit our analysis by considering only Watson-Crick complementary base pairs and G-U wobble base pairs. We compare the results of our statistical survey with existing theoretical and numerical models. We also discuss possible applications of this classification and show how it can be used for identifying new RNA structural motifs.