Triterpene saponin hemi-biosynthesis of a leaf beetle’s (<Emphasis Type="Italic">Platyphora kollari</Emphasis>) defensive secretion

The adults of the leaf beetle Platyphora kollari (Chrysomelidae) are able to metabolise the oleanane triterpene β-amyrin (1) into the glycoside 3-O-β-d-glucopyranosyl-(1→4)-β-d-glucuronopyranosyl-hederagenin (2) that is stored in their defensive glands. The aim of this study was to test the hypothesis that oleanolic acid (3) is an intermediate in the conversion of 1 into 2 and to check whether the sequestration of pentacyclic triterpenes is selective in favour of β-amyrin (1). To this end, adults of P. kollari were fed with Ipomoea batatas leaf disks painted with a solution of [2,2,3-²H₃]oleanolic acid or [2,2,3-²H₃]α-amyrin and the secretion of their defensive glands analysed by HPLC–ESIMS. The data presented in this work indicated that the first step of the transformation of β-amyrin (1) into the sequestered glycoside 2 is its oxidation into oleanolic acid (3) and that this conversion is selective but not specific in favour of β-amyrin (1).     

Brain modulation of Dufour’s gland ester biosynthesis in vitro in the honeybee (<Emphasis Type="Italic">Apis mellifera</Emphasis>)

Caste-specific pheromone biosynthesis is a prerequisite for reproductive skew in the honeybee. Nonetheless, this process is not hardwired but plastic, in that egg-laying workers produce a queen-like pheromone. Studies with Dufour’s gland pheromone revealed that, in vivo, workers’ gland biosynthesis matches the social status of the worker, i.e., sterile workers showed a worker-like pattern whereas fertile workers showed a queen-like pattern (production of the queen-specific esters). However, when incubated in vitro, the gland spontaneously exhibits the queen-like pattern, irrespective of its original worker type, prompting the notion that ester production in workers is under inhibitory control that is queen-dependent. We tested this hypothesis by exposing queen or worker Dufour’s glands in vitro to brain extracts of queens, queenright (sterile) workers and males. Unexpectedly, worker brain extracts activated the queen-like esters biosynthesis in workers’ Dufour’s gland. This stimulation was gender-specific; queen or worker brains demonstrated a stimulatory activity, but male brains did not. Queen gland could not be further stimulated. Bioassays with heated and filtered extracts indicate that the stimulatory brain factor is below 3,000 Da. We suggest that pheromone production in Dufour’s gland is under dual, negative–positive control. Under queenright conditions, the inhibitor is released and blocks ester biosynthesis, whereas under queenless conditions, the activator is released, activating ester biosynthesis in the gland. This is consistent with the hypothesis that queenright workers are unequivocally recognized as non-fertile, whereas queenless workers try to become “false queens” as part of the reproductive competition.     

Is microhabitat segregation between two cicada species (<Emphasis Type="Italic">Tibicina haematodes</Emphasis> and<Emphasis Type="Italic"> Cicada orni</Emphasis>) due to calling song propagation constraints?

The cicada species Tibicina haematodes and Cicada orni are two sympatric species often inhabiting vineyards. We show that they occupy two distinct levels: males of T. haematodes produce their calling songs from a high position in vine foliage while males of C. orni call from a low position near the ground on vine trunks. Experiments consisting of broadcasting and re-recording experimental signals in natural habitats from low and high positions show that signals are more and more modified as sender–receiver distance increases. T. haematodes would have an advantage when calling on trunks rather than on branches whereas C. orni would be able to call indiscriminately from both low and high positions. Thus, the microhabitat segregation observed between T. haematodes and C orni in vineyards does not seem to be related to calling song propagation constraints, but may be due to other ethological or ecological factors.     

Does a parthenogenesis-inducing <Emphasis Type="Italic">Wolbachia</Emphasis> induce vestigial cytoplasmic incompatibility?

Wolbachia is a maternally inherited bacterium that manipulates the reproduction of its host. Recent studies have shown that male-killing strains can induce cytoplasmic incompatibility (CI) when introgressed into a resistant host. Phylogenetic studies suggest that transitions between CI and other Wolbachia phenotypes have also occurred frequently, raising the possibility that latent CI may be widespread among Wolbachia. Here, we investigate whether a parthenogenesis-inducing Wolbachia strain can also induce CI. Parthenogenetic females of the parasitoid wasp Asobara japonica regularly produce a small number of males that may be either infected or not. Uninfected males were further obtained through removal of the Wolbachia using antibiotics and from a naturally uninfected strain. Uninfected females that had mated with infected males produced a slightly, but significantly more male-biased sex ratio than uninfected females that had mated with uninfected males. This effect was strongest in females that mated with males that had a relatively high Wolbachia titer. Quantitative PCR indicated that infected males did not show higher ratios of nuclear versus mitochondrial DNA content. Wolbachia therefore does not cause diploidization of cells in infected males. While these results are consistent with CI, other alternatives such as production of abnormal sperm by infected males cannot be completely ruled out. Overall, the effect was very small (9%), suggesting that if CI is involved it may have degenerated through the accumulation of mutations.     

1-Tridecene—male-produced sex pheromone of the tenebrionid beetle <Emphasis Type="Italic">Parastizopus transgariepinus</Emphasis>

Males of the genus Parastizopus (Coleoptera: Tenebrionidae) exhibit a special pheromone-emitting behaviour. They do a headstand, expose the aedeagus and remain in this posture for a few seconds. The pheromone emitted by P. transgariepinus was collected by solid-phase micro-extraction (100 microm polydimethylsiloxane fibre) and identified as 1-tridecene by gas chromatography/mass spectrometry. Presumably, this compound originates from the aedeagal gland, a special feature in Parastizopus, as 1-tridecene is the main compound in the gland reservoirs (23.6+/-3.8%), accompanied by various less volatile fatty acid esters (25.2+/-2.0%) and hydrocarbons (51.2+/-5.7%). 1-Tridecene is also part of the pygidial defensive secretion of both sexes, together with other 1-alkenes, monoterpene hydrocarbons and 1,4-benzoquinones, but as none of these other compounds was detected during calling, the pygidial gland could be ruled out as pheromone source. Extracts of the aedeagal gland reservoirs and the pygidial defensive secretion contained comparable amounts of 1-tridecene, 1.24+/-0.41 and 1.88+/-0.54 microg/male, respectively. Chemo-orientation experiments using a servosphere showed that 1 microg of 1-tridecene was attractive to females but not to males.     

The functionality of female reciprocal calls in the Iberian midwife toad (<Emphasis Type="Italic">Alytes cisternasii</Emphasis>): female-female acoustic competition?

Female midwife toads (genus Alytes) emit highly variable reciprocal calls of unclear function prior to and during courtship. In some species, female-female competition, expressed as physical fighting, has been reported. Males of Majorcan midwife toads (Alytes muletensis) show phonotactic response to female calls, and females of Iberian midwife toads (Alytes cisternasii) respond differently according to the male call characteristics. In this study, I test the hypothesis of female-female acoustic competition as an additional function of female reciprocal calls. Playback tests indicate that female calls are not clearly involved in female acoustic competition in the Iberian midwife toad, therefore female calls could be directed at males rather than towards competitive females.     

Parent–offspring communication in the Nile crocodile <Emphasis Type="Italic">Crocodylus niloticus</Emphasis>: do newborns’ calls show an individual signature?

Young Nile crocodiles Crocodylus niloticus start to produce calls inside the egg and carry on emitting sounds after hatching. These vocalizations elicit maternal care and influence the behaviour of other juveniles. In order to investigate the acoustic structure of these calls, focusing on a possible individual signature, we have performed acoustic analyses on 400 calls from ten young crocodiles during the first 4 days after hatching. Calls have a complex acoustic structure and are strongly frequency modulated. We assessed the differences between the calls of the individuals. We found a weak individual signature. An individual call-based recognition of young by the mother is thus unlikely. In other respects, the call acoustic structure changes from the first to the fourth day after hatching: fundamental frequency progressively decreases. These modifications might provide important information to the mother about her offspring—age and size—allowing her to customize her protective care to best suit the needs of each individual. Electronic supplementary material  Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Firewalls in bee nests—survival value of propolis walls of wild Cape honeybee (<Emphasis Type="Italic">Apis mellifera capensis</Emphasis>)

The Cape bee is endemic to the winter rainfall region of South Africa where fires are an integral part of the ecology of the fynbos (heathland) vegetation. Of the 37 wild nests in pristine Peninsula Sandstone Fynbos in the Cape Point section of Table Mountain National Park that have been analyzed so far, only 22 could be accessed sufficiently to determine the existence of a propolis wall of which 68% had propolis walls which entirely enclosed their openings. The analysis of the 37 wild nests revealed that 78% occurred under boulders or in clefts within rocks, 11% in the ground, 8% in tree cavities, and 3% within shrubs. The analysis of 17 of these nests following a fire within the park revealed that the propolis walls materially protected the nests and retarded the fire with all the colonies surviving. The bees responded to the smoke by imbibing honey and retreating to the furthest recess of their nest cavity. The bees were required to utilize this honey for about 3 weeks after which fire-loving plants appeared and began flowering. Considerable resources were utilized in the construction of the propolis walls, which ranged in thickness from 1.5 to 40 mm (mean 5 mm). Its physical environment determines the nesting behavior of the Cape bee. The prolific use of propolis serves to insulate the nest from extremes of temperature and humidity, restricts entry, camouflages the nest, and acts as an effective fire barrier protecting nests established mostly under rocks in vegetation subjected to periodic fires.     

The end of the fat dodo? A new mass estimate for <Emphasis Type="Italic">Raphus cucullatus</Emphasis>

A new mass estimate for the dodo (Raphus cucullatus), based on the lengths of the femur, tibiotarsus and tarsometatarsus, is attempted. The obtained mean mass is 10.2 kg, which is less than previous estimates based on other methods, which ranged from 10.6 to 21.1 kg, and much lower than the 50 lbs reported by a seventeenth-century eyewitness. The new estimated mass, which is similar to that of a large wild turkey, seems more realistic than previous ones and supports the hypothesis that contemporary illustrations of extremely fat dodos were either exaggerations, or based on overfed specimens. Pictures of “fat” dodos may also have been based on individuals exhibiting a display behaviour with puffed out feathers.     

A salamander’s toxic arsenal: review of skin poison diversity and function in true salamanders,genus <Emphasis Type="Italic">Salamandra</Emphasis>

Terrestrial salamanders of the genus Salamandra represent one of the most prominent groups of amphibians. They are mainly distributed across Europe but also reach Northern Africa and the Near East. Members of the six currently accepted species have long been known to be poisonous; however, work on their toxins was mostly published in German language, and therefore, many nuances of these studies have remained hidden from the majority of herpetologists and toxinologists. Several Salamandra species are called fire salamanders due to their highly contrasted, black-yellow colouration which probably serves to deter predators, although thorough evidence for aposematism in Salamandra is still lacking. Salamandra skin toxins do not only represent a potent antipredator defence but may also have antimicrobial effects. A better understanding of this dual function of Salamandra skin secretions is of utmost importance in the face of the emergence of a fungal disease causing catastrophic declines of fire salamanders in Central Europe, caused by the fungus Batrachochytrium salamandrivorans. In this review, we summarize the knowledge on Salamandra toxins, providing a list of the compounds so far isolated from their secretion and focusing on the bioactivity of the major compounds in Salamandra secretions, the steroidal alkaloids. We identify priorities for future research, including a screening of co-occurrence of steroidal alkaloids and tetrodotoxins in salamandrids, chemical characterization of already identified novel steroidal compounds, elucidation of the presence and role of peptides and proteins in the secretion, and experimental in vitro and in vivo study of the interactions between bioactive compounds in Salamandra skin secretions and cutaneous fungal and bacterial pathogens.     

Within-population variation in ejaculate characteristics in a prolonged breeder,Peron’s tree frog, <Emphasis Type="Italic">Litoria peronii</Emphasis>

Sperm number is often a good predictor of success in sperm competition; however, it has become increasingly clear that, for some species, variation in probability of paternity cannot be explained by sperm number alone. Intraspecific variation in ejaculate characteristics, such as the number of viable sperm and sperm longevity, may play an equally important role in determining fertilization success. Here, we assess variation among ejaculates in three factors that may contribute to fertilization success (number of sperm per ejaculate, viability, and longevity), in a population of Peron’s tree frog (Litoria peronii). We detected large variation among males in the number of sperm per ejaculate and the proportion of viable sperm within ejaculates, which could not be explained by variation in either male size or body condition. However, the proportion of viable sperm released by males increased over the season. Finally, we assessed sperm longevity (proportion viable sperm determined using a dual-fluorochrome vital dye) at two different temperatures. At 23°C, on average, 75% of sperm remained viable after 2 h, but there were significant differences amongst males with the percentage of viable sperm ranging from 43% to 95%. For sperm incubated at 4°C, ejaculates varied fivefold in sperm longevity with some males having 50% viable sperm after 5 days. Our data suggest that ejaculate characteristics (sperm number, viability, and longevity) vary widely in Peron’s tree frog and may therefore play an important role in determining siring success both in the presence and absence of sperm competition. We discuss the results in relation to selection on ejaculate traits via natural and sexual selection in this and other amphibians.     

Females’ sampling strategy to comparatively evaluate prospective mates in the peacock blenny <Emphasis Type="Italic">Salaria pavo</Emphasis>

Emerging evidence suggests the occurrence of comparative decision-making processes in mate choice, questioning the traditional idea of female choice based on rules of absolute preference. In such a scenario, females are expected to use a typical best-of-n sampling strategy, being able to recall previous sampled males based on memory of their quality and location. Accordingly, the quality of preferred mate is expected to be unrelated to both the number and the sequence of female visits. We found support for these predictions in the peacock blenny, Salaria pavo, a fish where females have the opportunity to evaluate the attractiveness of many males in a short time period and in a restricted spatial range. Indeed, even considering the variability in preference among females, most of them returned to previous sampled males for further evaluations; thus, the preferred male did not represent the last one in the sequence of visited males. Moreover, there was no relationship between the attractiveness of the preferred male and the number of further visits assigned to the other males. Our results suggest the occurrence of a best-of-n mate sampling strategy in the peacock blenny.     

Is dauer pheromone of<Emphasis Type="Italic"> Caenorhabditis elegans</Emphasis> really a pheromone?

Animals respond to signals and cues in their environment. The difference between a signal (e.g. a pheromone) and a cue (e.g. a waste product) is that the information content of a signal is subject to natural selection, whereas that of a cue is not. The model free-living nematode Caenorhabditis elegans forms an alternative developmental morph (the dauer larva) in response to a so-called dauer pheromone, produced by all worms. We suggest that the production of dauer pheromone has no fitness advantage for an individual worm and therefore we propose that dauer pheromone is not a signal, but a cue. Thus, it should not be called a pheromone.     

Does exaggerated morphology preclude plasticity to cuckoldry in the midshipman fish (<Emphasis Type="Italic">Porichthys notatus</Emphasis>)?

In species with more than one male reproductive morph, there typically exists a larger morph with exaggerated secondary sexual characters, and a smaller morph with reduced secondary sexual characters. These exaggerated and reduced morphologies are commonly thought to represent specializations to alternative behavioral reproductive tactics—large body size and exaggerated secondary sexual characters should both facilitate territoriality, courtship, and pair-spawning; while small body size and reduced secondary sexual characters should facilitate sneaky cuckoldry. Given this postulated relationship between morphology and behavior, we examined the relationship between the morphology of exaggerated males and cuckoldry. In a field and aquarium study of the midshipman fish, a fish with both exaggerated and reduced morphs, we demonstrated cuckoldry in some males of the exaggerated morph. Since the reduced morphology is thought to be an adaptation towards sneaky cuckoldry, we predicted that, of males with the exaggerated morph, less-exaggerated (smaller) males would be better able to gain proximity to the spawning pair during cuckoldry. In contrast to that prediction, access to the spawning pair during cuckoldry increased with the body size of the cuckolding exaggerated-morph males. This may be related to our observation that exaggerated males often cuckolded aggressively. Thus the exaggerated morphology need not preclude adaptive plasticity to cuckoldry, and may even aid it.     

Olfaction in a viscous environment: the “color” of sexual smells in <Emphasis Type="Italic">Temora longicornis</Emphasis>

We investigate chemical aspects of mating in the marine copepod Temora longicornis (Copepoda, Calanoidea). Our emphasis is the female pheromone signaling in form of well-defined trails for males to follow, observed in Doall et al. (Phil Trans R Soc Lond B 353:681–689, 1998). The viscous environment and the properties of the odorants play important roles as the spread of the pheromone trail limits the time during which it is useful for tracing. A key observation from our earlier work is the ability of a searching male to detect the direction of the female and to correct its swimming direction if necessary. We propose a simple mathematical model for the spread of a pheromone from a moving source and carry out numerical simulations of two possible detection mechanisms. We find that a searching agent that is capable to detect a ratio outperforms a searcher that depends on the gradient of a single compound. This suggests that copepod sex pheromones consist of blends of chemical compounds, and that a ratio detection mechanism similar to that in airborne insects is at work.     

A male gift to its partner? Cyanogenic glycosides in the spermatophore of longwing butterflies (<Emphasis Type="Italic">Heliconius</Emphasis>)

Males of several insect species transfer nuptial gifts to females during mating, typically in the form of a protein-rich spermatophore. In chemically defended species, males could potentially enhance such a gift with chemicals that help protect the female, her eggs, or both. This was shown for lepidopteran species that accumulate pyrrolizidine alkaloids. Most Heliconius butterflies are presumably protected from predators by virtue of de novo synthesized and/or sequestered cyanogenic glycosides. Males of Heliconius species are known to transfer nutritional gifts to the females but whether defensive chemicals could also be transferred is not known. To ascertain whether transfer of cyanogens occurs, we dissected freshly mated females from nine different Heliconius species and analyzed spermatophores for cyanogenic glycosides. We found cyanogens in the spermatophores of all nine species. This is the first time cyanogenic glycosides are reported in the spermatophores of arthropods. We discuss the implications of these findings for Heliconius biology and for other cyanogenic insects as well. We suggest that chemically defended species commonly lace their nuptial gifts with defensive chemicals to improve gift quality.     

Did the Romans introduce the Egyptian mongoose (<Emphasis Type="Italic">Herpestes ichneumon</Emphasis>) into the Iberian Peninsula?

New finds of bones of the Egyptian Mongoose (Herpestes ichneumon), one from Portugal and one from Spain, were directly ¹⁴C dated to the first century AD. While the Portuguese specimen was found without connection to the Chalcolithic occupation of the Pedra Furada cave where it was recovered, the Spanish find, collected in the city of Mérida, comes from a ritual pit that also contained three human and 40 dog burials. The finds reported here show that the Egyptian mongoose, contrary to the traditional and predominant view, did not first arrive in the Iberian Peninsula during the Muslim occupation of Iberia. Instead, our findings are consistent with the hypothesis that the species was first introduced by the Romans, or at least sometime during the Roman occupation of Hispania. Therefore, radiocarbon dating of new archaeological finds of bones of the Egyptian Mongoose (Herpestes ichneumon) in the Iberian Peninsula push back the confirmed presence of the species in the region by approximately eight centuries, as the previously oldest dated record is from the ninth century. With these new dates, there are now a total of four ¹⁴C dated specimens of Egyptian mongooses from the Iberian Peninsula, and all of these dates fall within the last 2000 years. This offers support for the hypothesis that the presence of the species in Iberia is due to historical introductions and is at odds with a scenario of natural sweepstake dispersal across the Straits of Gibraltar in the Late Pleistocene (126,000–11,700 years ago), recently proposed based on genetic data.     

Prospective thinking in a mustelid? <Emphasis Type="Italic">Eira barbara</Emphasis> (Carnivora) cache unripe fruits to consume them once ripened

The ability of nonhuman animals to project individual actions into the future is a hotly debated topic. We describe the caching behaviour of tayras (Eira barbara) based on direct observations in the field, pictures from camera traps and radio telemetry, providing evidence that these mustelids pick and cache unripe fruit for future consumption. This is the first reported case of harvesting of unripe fruits by a nonhuman animal. Ripe fruits are readily taken by a variety of animals, and tayras might benefit by securing a food source before strong competition takes place. Unripe climacteric fruits need to be harvested when mature to ensure that they continue their ripening process, and tayras accurately choose mature stages of these fruits for caching. Tayras cache both native (sapote) and non-native (plantain) fruits that differ in morphology and developmental timeframes, showing sophisticated cognitive ability that might involve highly developed learning abilities and/or prospective thinking.     

Individual protein balance strongly influences δ<Superscript>15</Superscript>N and δ<Superscript>13</Superscript>C values in Nile tilapia,<Emphasis Type="Italic"> Oreochromis niloticus</Emphasis>

Although stable isotope ratios in animals have often been used as indicators of the trophic level and for the back-calculation of diets, few experiments have been done under standardized laboratory conditions to investigate factors influencing ¹⁵N and ¹³C values. An experiment using Nile tilapia [Oreochromis niloticus (L.)] was therefore carried out to test the effect of different dietary protein contents (35.4, 42.3, and 50.9%) on ¹⁵N and ¹³C values of the whole tilapia. The fish were fed the isoenergetic and isolipidic semi-synthetic diets at a relatively low level. ¹⁵N and ¹³C values of the lipid-free body did not differ between the fish fed the diets with different protein contents, but the trophic shift for N and C isotopes decreased with increasing protein accretion in the individual fish, for N from 6.5 to 4 and for C in the lipid-free body from 4 to 2.5. This is the first study showing the strong influence of the individual protein balance to the degree to which the isotopic signature of dietary protein was modified in tissue protein of fish. The extrapolation of the trophic level or the reconstruction of the diet of an animal from stable isotope ratios without knowledge of the individual physiological condition and the feeding rate may lead to erroneous results.