Increase in membrane thickness during development compensates for eggshell thinning due to calcium uptake by the embryo in falcons

We compared membrane thickness of fully developed eggs with those of non-developed eggs in different endangered falcon taxa. To our knowledge, membrane thickness variation during development has never been examined before in falcons or any other wild bird. Yet, the egg membrane constitutes an important protective barrier for the developing embryo. Because eggshell thinning is a general process that occurs during bird development, caused by calcium uptake by the embryo, eggs are expected to be less protected and vulnerable to breakage near the end of development. Thus, egg membranes could play an important protective role in the later stages of development by getting relatively thicker. We used linear mixed models to explore the variation in membrane thickness (n = 378 eggs) in relation to developmental stage, taxon, female age, mass and identity (73 females), egg-laying sequence (105 clutches) and the study zone. Our results are consistent with the prediction that egg membranes are thicker in fully developed eggs than in non-developed eggs, suggesting that the increase in membrane thickness during development may compensate for eggshell thinning. In addition, our data shown that thicker membranes are associated with larger, heavier and relatively wider eggs, as well as with eggs that had thinner eggshells. Egg-laying sequence, female age and the study zone did not explain the observed variation of membrane thickness in the falcon taxa studied. As we provide quantitative data on membrane thickness variation during development in falcons not subjected to contamination or food limitation (i.e. bred under captive conditions), our data may be used as a reference for studies on eggs from natural populations. Considering the large variation in membrane thickness and the multiple factors affecting on it and its importance in the protection of the embryo, we encourage other researchers to include measurements on membranes in studies exploring eggshell thickness variation.     

Male behaviors reveal multiple pherotypes within vine mealybug Planococcus ficus (Signoret) (Hemiptera; Pseudococcidae) populations

The vine mealybug (VM) females collected in Israel produce two sex pheromone compounds: lavandulyl senecioate (LS) and (S)-lavandulyl isovalerate (LI). The males display ambiguous behavior to LI: repulsion in the vineyard and attraction of laboratory-reared males. We addressed the question of individual male behavior, i.e., do males respond to both LS and LI, or might they display a distinct response to each of the two pheromone compounds. We compared male pherotype frequencies between wild-caught and laboratory-reared populations. Then, we examined the relationship between pherotype composition and male capture rates in pheromone traps. Finally, we addressed the heredity of the pherotypes. The Israeli VM populations contain nine different male pherotypes, as defined according to the male behavior to pheromone compounds. The studied Portuguese populations included five of the nine pherotypes; none of the Portuguese males were attracted to LI. It seems that the high frequency of males that were attracted to LI is related to dense VM populations. It is hypothesized that selection for the male pherotypes, I males, those that respond to LI, occur under high-density rearing conditions. This may result from shorter development times of males and females that produce more I male pherotypes. The lower relative frequency of trapping of males in LI-baited traps than expected from the percentage determined in a Petri dish arena suggests that males that respond solely to LS (S males) are better fliers. The results also suggest that the pherotype trait is inherited by both sexes of the VM.     

A temporary social parasite of tropical plant-ants improves the fitness of a myrmecophyte

Myrmecophytes offer plant-ants a nesting place in exchange for protection from their enemies, particularly defoliators. These obligate ant–plant mutualisms are common model systems for studying factors that allow horizontally transmitted mutualisms to persist since parasites of ant–myrmecophyte mutualisms exploit the rewards provided by host plants whilst providing no protection in return. In pioneer formations in French Guiana, Azteca alfari and Azteca ovaticeps are known to be mutualists of myrmecophytic Cecropia (Cecropia ants). Here, we show that Azteca andreae, whose colonies build carton nests on myrmecophytic Cecropia, is not a parasite of Azteca–Cecropia mutualisms nor is it a temporary social parasite of A. alfari; it is, however, a temporary social parasite of A. ovaticeps. Contrarily to the two mutualistic Azteca species that are only occasional predators feeding mostly on hemipteran honeydew and food bodies provided by the host trees, A. andreae workers, which also attend hemipterans, do not exploit the food bodies. Rather, they employ an effective hunting technique where the leaf margins are fringed with ambushing workers, waiting for insects to alight. As a result, the host trees’ fitness is not affected as A. andreae colonies protect their foliage better than do mutualistic Azteca species resulting in greater fruit production. Yet, contrarily to mutualistic Azteca, when host tree development does not keep pace with colony growth, A. andreae workers forage on surrounding plants; the colonies can even move to a non-Cecropia tree.     

High-field 1H T1 and T2 NMR relaxation time measurements of H2O in homeopathic preparations of quartz, sulfur, and copper sulfate

Quantitative meta-analyses of randomized clinical trials investigating the specific therapeutic efficacy of homeopathic remedies yielded statistically significant differences compared to placebo. Since the remedies used contained mostly only very low concentrations of pharmacologically active compounds, these effects cannot be accounted for within the framework of current pharmacology. Theories to explain clinical effects of homeopathic remedies are partially based upon changes in diluent structure. To investigate the latter, we measured for the first time high-field (600/500 MHz) ¹H T₁ and T₂ nuclear magnetic resonance relaxation times of H₂O in homeopathic preparations with concurrent contamination control by inductively coupled plasma mass spectrometry (ICP-MS). Homeopathic preparations of quartz (10c–30c, n = 21, corresponding to iterative dilutions of 100⁻¹⁰–100⁻³⁰), sulfur (13x–30x, n = 18, 10⁻¹³–10⁻³⁰), and copper sulfate (11c–30c, n = 20, 100⁻¹¹–100⁻³⁰) were compared to n = 10 independent controls each (analogously agitated dilution medium) in randomized and blinded experiments. In none of the samples, the concentration of any element analyzed by ICP-MS exceeded 10 ppb. In the first measurement series (600 MHz), there was a significant increase in T₁ for all samples as a function of time, and there were no significant differences between homeopathic potencies and controls. In the second measurement series (500 MHz) 1 year after preparation, we observed statistically significant increased T₁ relaxation times for homeopathic sulfur preparations compared to controls. Fifteen out of 18 correlations between sample triplicates were higher for controls than for homeopathic preparations. No conclusive explanation for these phenomena can be given at present. Possible hypotheses involve differential leaching from the measurement vessel walls or a change in water molecule dynamics, i.e., in rotational correlation time and/or diffusion. Homeopathic preparations thus may exhibit specific physicochemical properties that need to be determined in detail in future investigations. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Cladistic biogeographic analysis suggests an early Caribbean diversification in Mexico

The Great American Biotic Interchange has been the predominant paradigm for explaining biotic diversification in the Nearctic/Neotropical overlap or Mexican Transition Zone, which is commonly explained by the collision of the North and South American continental plates, which began in the Oligocene and fused both landmasses. In the most far-reaching cladistic biogeographical analysis of the area to date, evidence has been found supporting the existence of a remnant Caribbean region extending from eastern Mexico to southeastern USA, a hypothesis that challenges current views of the Great American Biotic Interchange and the Mexican Transition Zone. We show herein that an older terrane, which has drifted to the present day positions of Yucatan and Cuba, may be biogeographically linked to an early ‘Gondwanan’ biota of the Paleocene (ca. 60 Ma). The evidence indicates an east–west biotic divide in Mexico, existing before the collision and formation of Central America. The south–north division of the country, previously recognized by several authors as associated with the Great American Biotic Interchange and the Mexican Transition Zone, is of a younger age. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Insecticide resistance may enhance the response to a host-plant volatile kairomone for the codling moth, <Emphasis Type="Italic">Cydia pomonella</Emphasis> (L.)

The behavioral and electroantennographic responses of Cydia pomonella (L.) to the ripe pear volatile ethyl (2E,4Z)-2,4-decadienoate (Et-E,Z-DD), were compared in insecticide-susceptible and -resistant populations originating from southern France. A dose–response relationship to this kairomonal attractant was established for antennal activity and did not reveal differences between susceptible and resistant strains. Conversely, males of the laboratory strains expressing metabolic [cytochrome P450-dependent mixed-function oxidases (mfo)] or physiological (kdr-type mutation of the sodium-channel gene) resistance mechanisms exhibited a significantly higher response to Et-E,Z-DD than those of the susceptible strain in a wind tunnel experiment. No response of the females to this kairomone could be obtained in our wind-tunnel conditions. In apple orchards, mfo-resistant male moths were captured at significantly higher rates in kairomone-baited traps than in traps baited with the sex pheromone of C. pomonella. Such a differential phenomenon was not verified for the kdr-resistant insects, which exhibited a similar response to both the sex pheromone and the kairomonal attractant in apple orchards. Considering the widespread distribution of metabolic resistance in European populations of C. pomonella and the enhanced behavioral response to Et-E,Z-DD in resistant moths, the development of control measures based on this kairomonal compound would be of great interest for the management of insecticide resistance in this species.     

Environmental assessment of propionic acid produced in an agricultural biomass-based biorefinery system

This paper presents an environmental system study of the production of propionic acid in a biorefinery system based on agricultural by-products. Here, propionic acid is produced by fermentation of glycerol, a by-product from biodiesel production, as carbon source and potato juice, a by-product from starch production, as nitrogen source. Biomass-based propionic acid leads to a greenhouse gas reduction of 60% compared to propionic acid from fossil sources. However, the primary energy input is about twice as high for the biomass-based propionic acid. The choice of input energy and the efficiency of the process, as well as the technology applied, have high impacts on the environmental performance of the biorefinery concept. There is a potential for increased integration both on the input of substrates and the output of end products of the biorefinery system, which will further improve its environmental performance.     

Isolation and characterization of facultative mixotrophic ammonia-oxidizing bacteria from constructed wetlands

Autotrophic ammonia-oxidizing bacteria (AOB) have been widely studied in constructed wetlands systems, while mixotrophic AOB have been less thoroughly examined. Heterotrophic bacteria were isolated from wastewater and rhizospheres of macrophytes of constructed wetlands, and then cultivated in a mixotrophic medium containing ammonium and acetic acid. A molecular characterization was accomplished using ITS-PCR amplification, and phylogenetic analysis based on 16S rRNA gene sequences. Results showed the presence of 35 bacteria, among 400 initially heterotrophic isolates, that were able to remove ammonia. These 35 isolates were classified into 10 genetically different groups based on ITS pattern. Then, a collection of 10 isolates were selected because of their relatively high ammonia removal efficiencies (ARE ≥ 80%) and their phylogenetic diversity. In conditions of mixotrophy, these strains were shown to be able to grow (increase of optical density OD₆₆₀ during incubation with assimilation of nitrogen into cellular biomass) and to oxidize ammonia (important ammonia oxidation efficiencies, AOE between 79% and 87%). Among these facultative mixotrophic AOB, four isolates were genetically related to Firmicutes (Bacillus and Exiguobacterium), three isolates were affiliated to Actinobacteria (Arthrobacter) and three other isolates were associated with Proteobacteria (Pseudomonas, Ochrobactrum and Bordetella).     

Alkaloid concentration of the invasive plant species Ulex europaeus in relation to geographic origin and herbivory

In the study of plant defense evolution, invasive plant species can be very insightful because they are often introduced without their enemies, and traits linked to defense can be released from selective pressures and evolve. Further, studying plant defense evolution in invasive species is important for biological control and use of these species. In this study, we investigated the evolution of the defensive chemicals quinolizidine alkaloids (QAs) in the invasive species gorse, Ulex europaeus. Using a common garden experiment, our goals were to characterize the role of QAs relative to specialist enemies of gorse and to investigate if QA concentration evolved in invaded regions, where gorse was introduced without these enemies. Our results showed that pod infestation rate by the seed predator Exapion ulicis and infestation by the rust pathogen Uromyces genistae-tinctoriae were negatively correlated to concentration of the QA lupanine. Quinolizidine alkaloid concentration was very variable between individuals, both within and among populations, but it was not different between native and invaded regions, suggesting that no evolution of decreased resistance occurred after gorse lost its enemies. Our study also suggests that QA concentrations are traits integrated into seed predation avoidance strategies of gorse, with plants that mass-fruit in spring but do not escape pod infestation in time being richer in QAs.     

Uncommon formation of two antiparallel sperm bundles per cyst in tenebrionid beetles (Coleoptera)

Several species of Tenebrionidae are stored-grain pests. Since they belong to a specious family, the systematics of these beetles is still in doubt. In insects, spermatogenesis and the spermatozoa exhibit great diversity, and are therefore commonly used in phylogenetic and taxonomic analyses. During the spermatogenetic process in these organisms, the cells originating from a single spermatogonium develop synchronically in groups referred to as cysts. At the end of this process, there is usually only one sperm bundle per cyst, with all the cells in the same orientation. This paper details the spermiogenesis of the tenebrionid beetles Tenebrio molitor, Zophobas confusa, Tribolium castaneum and Palembus dermestoides using whole mount and histological sections of the cysts. In these species, spermatogenesis is similar to that which occurs in most insects. However, during spermiogenesis, the nuclei of the spermatids migrate to two opposite regions at the periphery of the cyst, leading to the uncommon formation of two bundles of spermatozoa per cyst. This feature is possibly an apomorphy for Tenebrionidae.