Salinity favorable for early development and gamete compatibility in two sympatric estuarine species of the genus Hediste (Polychaeta: Nereididae) in the Ariake Sea, Japan

Early development was examined, under various salinities, for two sympatric nereidid polychaetes, Hediste japonica and H. diadroma, which participated in a simultaneous reproductive swarming in an estuary of the Omuta-gawa River in Ariake Sea, Japan. The eggs of both species were isotonic to the medium of 27.5–30 psu salinity. The egg diameter in the isotonic salinity was 180–205 μm in H. japonica, and 130–160 μm in H. diadroma. Successful development of most embryos was observed in a salinity range of 22.5–30 psu in both species, while successful fertilization occurred in wider ranges of salinity, i.e., 10–34 psu in H. japonica and 10 to 30 psu in H. diadroma. In both species, free-swimming larval life started from the indistinct hatching of trochophores out of a jelly layer capsule. The lecithotrophic development appeared to run to the 4-setiger nectochaetes in H. japonica, while to 3-setiger nectochaetes in H. diadroma, resulting in a shorter pelagic larval life in H. japonica. In a comparison of larval morphology among Hediste species, we found a definite negative correlation between the prostomium width, which represents the larval size and depends on egg size, and relative length of chaetae to the prostomium width: the relative length of chaetae was the longest in H. diadroma (with the smallest egg size and long pelagic life), intermediate in H. japonica (intermediate egg size, short pelagic life), and the shortest in H. atoka (largest egg size, no true pelagic life). We also examined the possibility of hybridization between H. japonica and H. diadroma through cross-insemination experiments. The gametes of the two species were reciprocally compatible, and viable hybrid offspring were produced by the laboratory crosses. The hybrid larvae expressed intermediate phenotypes, but with a greater maternal influence in characteristics such as the relative length of chaetae and the lecithotrophic larval duration.     

Rhythmicity of larval release in three species of intertidal brachyuran crabs (Crustacea: Brachyura) from Inhaca Island (Mozambique)

 A study of rhythmicity of larval release in three species of intertidal brachyuran crabs, based on laboratory and field experiments, was undertaken at Inhaca Island, southern Mozambique, using Leptodius exaratus and Macrophthalmus grandidieri from December 1994 to January 1995, and Arcotheres palaensis from April to July 1995. L. exaratus and M. grandidieri showed a semi-lunar cycle in larval release. The release of larvae for L. exaratus, a species having conspicuous larvae, occurred in the first half of the night, after the post-crepuscular high tide, which suggests maximisation of protection of larvae from visual predation. The larval release activity matched the late spring and early neap tides. Results from the field were similar to those from the laboratory. M. grandidieri, having inconspicuous larvae, did not show a pattern related to the light–dark cycle and hatched during spring tides (around full and new moons) to maximise larval dispersion. A. palaensis, living inside the host mussel which inhabits the lowest section of the intertidal zone, did not show a relation with moon phase, tidal or light–dark cycles. Received: 16 February 1999 / Accepted: 8 December 1999     

Pheromones and colonization: reassessment of the milkweed bug migration model (Heteroptera: Lygaeidae: Lygaeinae)

Summary. Research on insect migration has justifiably emphasized females – the so-called “oogenesis-flight syndrome”– since it is the females that place the eggs into new habitats. The large and small milkweed bugs, Oncopeltus fasciatus and Lygaeus kalmii, respectively, have featured prominently in studies of insect migration and sequestration of host plant toxins for chemical defense. Here we report that males of these species, and males of another well-studied lygaeine (Neacoryphus bicrucis), produce pheromones in glands usually considered to serve only a defensive role in Heteroptera (the metathoracic scent glands), and that these pheromones are exploited by a tachinid parasitoid as a host-finding kairomone. The pheromones are mixtures of C₆ and C₈ saturated and unsaturated esters reminiscent of lepidopteran pheromones, and the key compound of the O. fasciatus pheromone has now been correctly identified as (E)-2,7-octadienyl acetate. It is proposed that the concept of the oogenesis-flight syndrome for these kinds of insects should accommodate the role of males in the migration process. The hypothesis is presented that male-produced pheromones play a significant role in guiding colonization of new habitats in many heteropteran species. In addition, data are presented suggesting that there is a trade-off between the amount of pheromone produced by colonizing males and the host breadth of the species. Received 21 December 1998; accepted 15 February 1999.     

Swimming ability and its rapid decrease at settlement in wrasse larvae (Teleostei: Labridae)

Wrasses are abundant reef fishes and the second most speciose marine fish family, yet little is known of their larval swimming abilities. In August 2010 at Moorea, Society Islands, we measured swimming ability (critical speed, Ucrit) of 80 settlement-stage larvae (11–17 mm) of 5 labrid species (Thalassoma quinquevittatum [n = 67], Novaculichthys taeniourus [n = 6], Coris aygula [n = 5], Halichoeres trimaculatus [n = 1] and H. hortulanus [n = 1]) and 33 new recruits of T. quinquevittatum. Median (mdn) larval Ucrit was 7.6–12.5 cm s⁻¹. In T. quinquevittatum (n = 67), larvae of 12.5–14.5 mm swam faster (mdn 16.9 cm s⁻¹) than smaller or larger larvae (mdn 3.9 and 3.2 cm s⁻¹, respectively). Labrid larvae Ucrit is similar to that of other similar-sized tropical larvae, so labrids and species with comparable settlement sizes should have similar abilities to influence dispersal. Ucrit of T. quinquevittatum recruits decreased to 47–56% of larval Ucrit in 2 days, implying rapid physiological changes at settlement.     

Spawning and development in <Emphasis Type="Italic">Osedax</Emphasis> boneworms (Siboglinidae,Annelida)

We report observations on spawning and early development in bone-eating worms of the genus Osedax. Individual females of Osedax rubiplumus were observed at 1820 m depth freely spawning hundreds of oocytes, and females of an undescribed species, Osedax “orange collar”, were observed spawning in laboratory aquaria. Cytological and molecular analysis of the spawned oocytes of two Osedax species revealed no evidence for the bacterial endosymbionts that the female worms require for their nutrition, suggesting that the bacteria must be acquired later from the environment, as they are in other siboglinids. Individual O. “orange collar” females released an average of 335 (±130) eggs per day, but the number of oocytes spawned per day varied greatly, suggesting that not all the females spawned daily. Fertilization rates of the spawned oocytes varied from 0 to 100%, though most females showed nearly 100% fertilization rates. Oocytes spawned in the laboratory at 4–6°C were negatively buoyant. If fertilized, these oocytes extruded polar bodies and then after at least four hours cleaved unequally. Subsequent cleavages occurred in a spiral pattern at roughly 2-h intervals, resulting in free-swimming trochophore larvae after 24 h. These lecithotrophic trochophores swam for 9–16 days before settling with several hooked chaetae, similar to those of dwarf Osedax males. The larval life span of the Osedax species studied in the laboratory appears to be shorter than in closely related Vestimentifera. Osedax rubiplumus, on the other hand, has much larger oocytes and so may have greater dispersal potential than these other Osedax species. The high fecundity and apparently continuous reproduction of Osedax boneworms permits the opportunistic exploitation of sunken vertebrate bones. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Phylogeography of two Atlantic squirrelfishes (Family Holocentridae): exploring links between pelagic larval duration and population connectivity

Genetic surveys of reef fishes have revealed high population connectivity within ocean basins, consistent with the assumption that pelagic larvae disperse long distances by oceanic currents. However, several recent studies have demonstrated that larval retention and self-recruitment may be higher than previously expected. To assess connectivity in tropical reef fishes, we contribute range-wide mtDNA surveys of two Atlantic squirrelfishes (family Holocentridae). The blackbar soldierfish, Myripristis jacobus, has a pelagic juvenile phase of about 58 days, compared to about 71 days (~22% longer) in the longjaw squirrelfish, Holocentrus ascensionis. If the pelagic duration is guiding dispersal ability, M. jacobus should have greater population genetic structure than H. ascensionis. In comparisons of mtDNA cytochrome b sequences from 69 M. jacobus (744 bp) and 101 H. ascensionis (769 bp), both species exhibited a large number of closely related haplotypes (h=0.781 and 0.974, π=0.003 and 0.006, respectively), indicating late Pleistocene coalescence of mtDNA lineages. Contrary to the prediction based on pelagic duration, M. jacobus has much less population structure (φ_ST=0.008, P=0.228) than H. ascensionis (φ_ST=0.091, P<0.001). Significant population partitions in H. ascensionis were observed between eastern, central and western Atlantic, and between Brazil and the Caribbean in the western Atlantic. These results, in combination with the findings from 13 codistributed species, indicate that pelagic larval duration is a poor predictor of population genetic structure in Atlantic reef fishes. A key to understanding this disparity may be the evolutionary depth among corresponding taxonomic groups of “reef fishes”, which extends back to the mid-Cretaceous and encompasses enormous diversity in ecology and life history. We should not expect a simple relationship between pelagic larval duration and genetic connectivity, among lineages that diverged 50–100 million years ago.     

Storage and display of odour by male Saccopteryx bilineata (Chiroptera, Emballonuridae)

Males of the sac-winged bat, Saccopteryx bilineata, actively fill their propatagial sacs with secretions from the genital region, the gular gland, urine and saliva. From our observations and those of Starck we deduce that propatagial sacs in S. bilineata do not have a glandular function, but are instead organs for the storage and display of odours. In addition to the already known “salting” and hovering behaviour of male S. bilineata, we describe in detail how odour is fanned to roosting individuals during the complex, stereotypic hovering displays. S. bilineata males also coat the fur of their backs with saliva using the wing tip and might scent-mark territory boundaries. “Yawning” may represent a visual as well as an olfactory cue. Odour seems to play an important role in the social communication of S. bilineata and in other emballonurids, as revealed by the broad distribution of wing sacs in this family. S. bilineata males display odour during energetically costly hovering flights in front of females. We demonstrate that the number of hovering displays increases with harem size. The mating effort of S. bilineata males comprises a multimodal signalling behaviour. Although males defend harem territories in which females gather, females seem to be able to choose the father of their progeny freely among the males of a colony. This may have led to the evolution of the complex mating displays by male S. bilineata. Received: 9 December 1998 / Received in revised form: 6 May 1999 / Accepted: 13 June 1999     

Sex determination inScalpellum scalpellum (Cirripedia: Thoracica: Lepadomorpha), a hermaphroditic goose barnacle with dwarf males

The cyprid larvae ofScalpellum scalpellum (L.), a hermaphroditic goose barnacle with dwarf males, are morphologically alike and no characters distinguish the sexes. To elucidate the controversy over the relative significance of genetic and environmental factors in the sex determination of this species, laboratory experiments were performed involving the introduction of cyprid larvae of different parental origin to different densities of adult hermaphrodites without dwarf males. Cyprids introduced to adult hermaphrodites metamorphosed to either dwarf males or hermaphrodites significantly different from the controls without adult hermaphrodites. The proportion which metamorphosed into dwarf males increased with increasing adult density. Not more than 51% metamorphosis to dwarf males could be obtained, indicating a genetic component in the sex differentiation. It was concluded that all cyprids are potential hermaphrodites, but that about 50% possess the capability of metamorphosing into dwarf males as well, if adult hermaphrodites with free receptacles are available. Consequently, both genetically and environmentally controlled components influence sex determination in this species.     

Preservation and loss of the honey bee (<Emphasis Type="Italic">Apis</Emphasis>) egg-marking signal across evolutionary time

Honey bee workers are able to distinguish queen-laid eggs from worker-laid eggs, and remove (‘police’) worker-laid eggs. The cue that police workers use is as yet unidentified but is likely to be a chemical signal. This signal benefits queens for it ensures their reproductive monopoly. It also benefits collective workers because it allows them to raise more closely related queen-laid males than the less-related sons of half sisters. Because both parties benefit from the egg-marking signal, it should be stable over evolutionary time. We show that Apis mellifera workers can distinguish queen-laid from worker-laid eggs of the dwarf honey bee A. florea, a phylogenetically distant species that diverged from the A. mellifera lineage 6–10 mya. However, A. mellifera workers are unable to distinguish worker-laid eggs of A. cerana, a much more recent divergence (2–3 mya). The apparent change in the egg-marking signal used by A. cerana may be associated with the high rates of ovary activation in this species.     

Feeding by larvae of two different developmental modes in Streblospio benedicti (Polychaeta: Spionidae)

Females of the spionid polychaete Streblospio benedicti (Webster) produce either small eggs (60–70 μm diameter) and planktotrophic larvae, or large eggs (100–200 μm) and lecithotrophic larvae that reportedly do not feed. This intraspecific polymorphism, a form of poecilogony, is potentially useful in studies of larval ecology and evolution, but necessary data on larval form and function are lacking. This study describes the morphology and nutritional biology of larvae obtained from Atlantic (South Carolina) and Pacific (California and Washington) populations from 2003 to 2005. The two types of larvae produced by Atlantic S. benedicti differed greatly in length (229±22 μm SD for planktotrophs vs. 638±40 μm for lecithotrophs) and chaetiger number (2–5 vs. 10–11) at release from the female’s brood pouch. Planktotrophic larvae bore long provisional chaetae on their first chaetiger; provisional chaetae were absent in lecithotrophic larvae. Larvae from Pacific populations were all of the lecithotrophic form, and were similar to their Atlantic counterparts in all respects. High-speed video microscopy revealed that both types of larvae used opposed bands of cilia to capture suspended particles and transport them to the mouth, where they were often ingested. Lecithotrophic larvae reared with suspended phytoplankton (Rhodomonas sp., 10⁴ cells ml⁻¹) for 2 days grew significantly faster than sibling larvae reared without added food, indicating that these larvae can digest and assimilate ingested food. Larvae of S. benedicti that develop from large eggs are thus facultative planktotrophs instead of obligately non-feeding lecithotrophs, a result that affects the interpretation of comparative studies of the ecology and evolution of larvae in S. benedicti and certain other marine invertebrates.     

Invasion patterns inferred from cytochrome oxidase I sequences in three bryozoans, Bugula neritina, Watersipora subtorquata, and Watersipora arcuata

Nucleotide variation in cytochrome c oxidase subunit I (COI) was used to examine population structure in three invasive bryozoans: Bugula neritina (Linnaeus, 1758), Watersipora subtorquata (d’Orbigny, 1852), and W. arcuata (Banta, 1969). These species are found on ship hulls and have a short (≤2 days) larval phase. Samples were collected from 1998–2001 at multiple sites in Australia, and in Hong Kong, New Zealand, Hawaii, California, Curaçao, and England. B. neritina is known to include three cryptic species, including species Type S (Davidson and Haygood in Biol Bull 196:273–280, 1999) which occurs on the east and west coasts of the USA. One haplotype recorded previously in the USA, S1, was found to be widespread, occurring throughout Australia and in Hong Kong, Curaçao, Hawaii, and England. W. subtorquata, a Caribbean–Atlantic species which has invaded southern Australia, New Zealand, and California, had low nucleotide diversity in these areas (π=0.0016±0.0014), consisting of three haplotypes connected by one or two nucleotide mutations. W. arcuata, an Eastern-Pacific native, had comparatively high diversity (π=0.0221±0.0115) in introduced populations from Australia and Hawaii. In each species, identical haplotypes were identified on separate coastlines providing evidence of widespread, rather than genetically independent, introductions. The major contrast in nucleotide diversity suggests that different propagule-source models explain introductions.     

Temporal variation of Tubularia indivisa (Cnidaria, Tubulariidae) and associated epizoites on artificial habitat communities in the North Sea

We have analyzed the composition, diversity, density and biomass of a temporal series of samples taken in a Tubularia indivisa community, which dominates a shipwreck in the North Sea waters (N 51°23′,730–E 02°29′,790, 17 nautical miles from the coast, 30 m depth). This shipwreck has structures emerging up to 8 m above the seabed. Water temperature ranged from 4.2°C in March to 20.3°C in August. Salinity showed few variations around 33.9 psu. Bottom tidal currents followed a semi-diurnal cycle and were preferentially NE oriented with 84% of them in the range 0.25–0.75 m s⁻¹. The mean value for total suspended matter was 6.2 mg l⁻¹ with large variations on a monthly scale. The species richness of samples varied from 15 in October to 42 in August with a mean value of 33 species. Diversity indices were higher during autumn and winter because of the strong dominance of a few crustacean species during the warmer months. The total density of individuals ranged from 6,500 ind m⁻² in October to 445,800 ind m⁻² in July, most of these individuals belonging to the amphipod species Jassa herdmani. The biomass of the T. indivisa community varied from 9 g AFDW m⁻² in October to 1,106 g AFDW m⁻² in July, with T. indivisa itself constituting between 59 and 82% of the total biomass. The biomass of T. indivisa was positively correlated with species richness and with the density of 23% of the species identified on this community, suggesting that T. indivisa plays an important structural role in this habitat. This was further confirmed by the number of species associated with T. indivisa which was generally superior to 55% of the sorted species. Multivariate analysis indicated strong differences between spring/summer−autumn/winter assemblages mostly but not solely due to the abundance patterns of species. These findings support the conclusion that shipwrecks in Belgian waters allow the development of assemblages dominated by a high biomass of T. indivisa which in turn provides shelter for high densities and biomass of epizoites. These assemblages will further show large monthly variations in densities and composition due to large variation in T. indivisa biomass under an apparent repetitive annual cycle. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Three party symbiosis: acoelomorph worms,corals and unicellular algal symbionts in Eilat (Red Sea)

Epizoic worms were found to occur on certain coral colonies from reefs off the coast of Eilat (Red Sea). We identified 14 coral species infested by acoelomorph worms at a depth range of 2–50 m. The host corals were all zooxanthellate and included both massive and branching stony corals and a soft coral. Worms from all hosts were identified as belonging to the genus Waminoa and contained two distinct algal symbionts differing in size. The smaller one was identified as Symbiodinium sp. and the larger one is presumed to belong to the genus Amphidinium. Worm-infested colonies of the soft coral, Stereonephthya cundabiluensis, lacked a mucus layer and exhibited distinct cell microvilli, a phenotype not present in colonies lacking Waminoa sp. In most cases, both cnidarian and Acoelomorph hosts displayed high specificity for genetically distinctive Symbiodinium spp. These observations show that the epizoic worms do not acquire their symbionts from the “host” coral.     

Alternative reproductive tactics and male-dimorphism in the horned beetle Onthophagus acuminatus (Coleoptera: Scarabaeidae)

Adult dung beetles (Onthophagus acuminatus) exhibit continuous variation in body size resulting from differential nutritional conditions experienced during larval development. Males of this species have a pair of horns that protrude from the base of the head, and the lengths of these horns are bimodally distributed in natural populations. Males growing larger than a threshold body size develop long horns, and males that do not achieve this size grow only rudimentary horns or no horns at all. Previous studies of other horned beetle species have shown that horned and hornless males often have different types of reproductive behavior. Here I describe the mating behaviors of the two male morphs of O. acuminatus during encounters with females. Females excavate tunnels beneath dung, where they feed, mate and provision eggs. Large, horned males were found to guard entrances to tunnels containing females. These males fought with all other males that attempted to enter these tunnels. In contrast, small, hornless males encountered females by sneaking into tunnels guarded by other males. In many instances, this was accomplished by digging new tunnels that intercepted the guarded tunnels below ground. Side-tunneling behavior allowed sneaking males to enter tunnels beneath the guarding male, and mate with females undetected. Both overall body size and relative horn length significantly affected the outcome of fights over tunnel ownership. These results suggest that alternative reproductive tactics may favor divergence in male horn morphology, with long horns favored in males large enough to guard tunnels, and hornlessness favored in smaller males that adopt the “sneaking” behavioral alternative. Received: 12 October 1996 / Accepted after revision: 8 August 1997     

Age determination in the Antarctic shrimp Notocrangon antarcticus (Crustacea: Decapoda), using the autofluorescent pigment lipofuscin

Determination of basic population parameters in long-lived Crustacea is hindered by a lack of appropriate methods for age determination. This study uses the pigment lipofuscin as an age marker in the common Antarctic decapod Notocrangon antarcticus (Pfeffer, 1887) from the eastern Weddell Sea. Resin brain sections of the lateral somacluster of the olfactory lobe of 189 specimens were digitally recorded by confocal microscopy and images were subsequently analyzed. A modal progression analysis of the lipofuscin concentration–frequency distribution revealed a total of eight regularly spaced modes presumed to reflect consecutive annual age classes. All eight modes contained females, and the first four contained males. No regular modes were obvious in the comparable length-frequency distribution. The average yearly pigment accumulation was nearly linear and estimated as 0.02% area fraction year⁻¹, which is considerably lower than rates published for species from lower latitudes. This is probably explained by the effect of low water temperature on metabolism and lipofuscin accumulation rate. The growth parameters CL_∞ and k from the von Bertalanffy growth function were 22.3 mm and 0.79 year⁻¹ for females, respectively, and 16.9 mm and 0.64 year⁻¹ for males, respectively. Mortality, estimated from catch curves, amounted to 0.44 year⁻¹ in females and 0.92 year⁻¹ in males, whereas P/B, calculated from the mass specific growth rate method, was slightly lower. The results indicate that the lipofuscin-inferred population parameters are an improvement over what can be learned about N. antarcticus with traditional methods. Received: 1 August 2000 / Accepted: 21 September 2000     

RAPD-PCR investigations of systematic relationships among four species of eels (Teleostei: Anguillidae), particularly Anguilla anguilla and A. rostrata

 We examined the genetic identities of 49 individuals of four species of eels in the genus Anguilla Shaw, A. anguilla, A. japonica, A. reinhardti and A. rostrata, using the random(ly) amplified polymorphic DNA (RAPD)-PCR technique. We used 15 random decamer primers. Out of the 22 148 amplification products detected, 454 markers were evaluated using Nei's distance coefficient, two numerical methods (neighbor-joining, UPGMA), and phylogenetic analysis using parsimony (PAUP). Both numerical and parsimony methods were congruent and confirmed the specific status of all four examined Anguilla species. In all dendrograms, all individuals of each of the species clustered together within four closed groups with highly significant bootstrap values (between 97 and 100%). Additionally 1 to 18 species-specific monomorphic diagnostic fragments were detected in three of the species; none were detected in A. anguilla. Mean interspecific genetic distances ranged from 0.384 to 0.559, with the minimum between A. anguilla and A. rostrata and the maximum between A. anguilla and A. reinhardti. Individuals collected at one locality and belonging to a single species did not cluster together, and neither were any diagnostic monomorphic fragments found for individuals of single localities. This probably reflects a random dispersal of larvae in ocean currents before they reach coastal waters. Among all possible species pairs, only the two Atlantic species were consistently clustered in all dendrograms, with highly significant bootstrap values (100%). Additionally, we detected ten diagnostic markers for this pair of eel species. From RAPD data, we suggest a phylogram which was routed by the descending analysis method: A. reinhardti appears to have been the first species to diverge from a putative common ancestor of the four eel species. Later, A. japonica separated from the hypothetical ancestor of the Atlantic species, which then split recently into A. anguilla and A. rostrata. The close relationship of A. anguilla and A. rostrata is indicated by their low genetic distance (GD = 0.384) and a high degree of shared RAPD fragments (SF = 71.2%); this compares with the following means for all other species pairs: ( = 0.531,  = 44.2%). The prerequisites for using RAPD data for the reconstruction of phylogenies are discussed. Received: 2 September 1999 / Accepted: 8 May 2000     

The genetic mating system of a sex-role-reversed pipefish (Syngnathus typhle): a molecular inquiry

In the pipefish Syngnathus typhle as in other species of Syngnathidae, developing embryos are reared on the male's ventral surface. Although much laboratory research has been directed toward understanding sexual selection in this sex-role-reversed species, few studies have addressed the mating behavior of S. typhle in the wild, and none has capitalized upon the power of molecular genetic assays. Here we present the first direct assessment of the genetic mating system of S. typhle in nature. Novel microsatellite loci were cloned and characterized from this species, and employed to assay entire broods from 30 pregnant, field-captured males. Genetic analysis of 1340 embryos revealed that 1–6 females (mean = 3.1) contributed to each brooded clutch, the highest rate of multiple maternity yet documented in any pipefish. Evidence of multiple mating by females was also detected. Thus, this population of S. typhle displays a polygynandrous mating system, a finding consistent with previous field and laboratory observations. Our results, considered together with similar studies of other syngnathid species, provide preliminary support for the hypothesis that the genetic mating system is related to the evolution of sexual dimorphism in the fish family Syngnathidae. Received: 19 January 1999 / Received in revised form: 15 April 1999 / Accepted: 9 May 1999     

Effect of host and larval frass volatiles on behavioural response of the old house borer,Hylotrupes bajulus (L.) (Coleoptera: Cerambycidae), in a wind tunnel bioassay

Summary. In a wind tunnel bioassay the effect of three concentrations of natural extracts of (1) Scots pine wood, Pinus sylvestris, and (2) larval frass on the behavioural response of unmated females and males of the old house borer, Hylotrupes bajulus, was tested and compared to the behavioural effects of the male-produced sex pheromone (3R)-3-hydroxy-2-hexanone. The influence on the behaviour of both sexes was found to be equally significant for the two higher concentrated hexane extracts of wood and larval frass. Therefore several synthetic monoterpenes present in the extracts and ethanol were tested at the two higher concentrations (1:100, 1:1000 vol/vol). Among the higher concentrated monoterpenoid hydrocarbons [(+)-α-pinene, (+)-β-pinene, (+)-limonene], only α-pinene increased the activity, orientation towards scent source and interest towards conspecifics. The tests with higher concentrated ethanol and the oxygenated monoterpenes [(-)-verbenone, (-)-trans-pinocarveol, (+)-terpinen-4-ol, (+)-α-terpineol, (-)-myrtenol] revealed that verbenone is the most effective stimulant for the females, followed by trans-pinocarveol, terpinen-4-ol and α-terpineol. For males, terpinen-4-ol was the only mediator significantly inducing attraction and orientation towards the scent source combined with an interest in conspecifics apparent by fighting or courtship behaviour. Males did not respond to verbenone which is a main compound of larval frass. Myrtenol and ethanol were ineffective in both sexes. In fact behavioural observations suggest that the beetles were repelled by the high dose of myrtenol. Using the ten-fold lower dose of the synthetic monoterpenes (1:1000 vol/vol), all semiochemicals except myrtenol lost activity. Myrtenol, however, induced behavioural responses, like increased activity and orientation towards scent source, only at the low concentration. Based on the results, primary attraction of unmated old house borer is probably mediated by monoterpenes of coniferous wood, while secondary attraction to infested wood would occur in response to volatiles of larval frass. Received 5 May 1999; accepted 30 September 1999     

Reproductive cycles and larval morphology of three Recent species of Argyrotheca (Terebratellacea: Brachiopoda) from Mediterranean submarine caves

Although the three examined species of Argyrotheca are quite common in the Mediterranean, little is known about their reproductive cycles. This study shows that Argyrotheca cordata and Argyrotheca cistellula have continuous breeding activity, indicated by the occurrence of ripe eggs and larvae in their brood pouches throughout the year. The absolute number of larvae found in the brood pouches of both species corresponds to the adult's size. The third species examined, Argyrotheca cuneata, tends to breed in autumn, with larvae only present in September and November, but the sample-size was small. Larval development is very similar in all three Argyrotheca species, each embryo going through a gastrula stage, a two-lobed and a three-lobed stage. Before leaving the brood pouch, the larvae have an apical lobe with a girdle of elongated cilia, a mantle lobe with a midventral band of cilia and a pedicle lobe without ciliation. The larvae of A. cordata and A. cuneata possess four bundles of larval setae, whereas A. cistellula shows no setae at all. Received: 6 November 1998 / Accepted: 15 March 1999     

Testing the ability of males and females to respond to altered songs in the dueting green lacewing, Chrysoperla plorabunda (Neuroptera: Chrysopidae)

Green lacewings in the carnea group of Chrysoperla engage in species-specific heterosexual duets using low-frequency substrate-borne signals. Within each species, both sexes sing nearly identical songs. Songs are the principal barriers to hybridization between sympatric species in the complex. Here, we investigated the responsiveness of males and females of Chrysoperla plorabunda to synthesized, prerecorded songs that differed from the species mean in the period between repeated volleys of abdominal vibration. We tested 15–16 males and 15–16 females using playbacks of two signals that gradually increased or decreased in volley period, starting at the species mean. We found that (1) duets during courtship are accurate, interactive, and adjustable by each participant; (2) in staged duets, both sexes respond best to song tempos near the mean volley period of their population, but can nonetheless maintain duets with signals of nearly twice, or half, the normal volley period; (3) individuals fine-tune their adjustments to signals of different volley periods by changing their own volley duration and latent period, or less often by inserting extra volleys or skipping every other volley; (4) males are significantly better at matching signals of changing tempo than females; and (5) the range of song responsiveness of C. plorabunda does not overlap the natural range of volley periods found in Chrysoperla adamsi, an acoustically similar sibling species, thus reaffirming strong behavioral isolation. In sum, the precise, almost unbreakable heterosexual duets characteristic of song species of the carnea group result from tight mutual feedback between partners. Effective reproductive isolation between species can be based on song differences alone.