Morphological process of vertical transmission of photosymbionts in the colonial ascidian <Emphasis Type="Italic">Trididemnum miniatum</Emphasis> Kott, 1977

In obligate symbioses between didemnid ascidians and prokaryotic algae (cyanobacteria), the larvae of the host ascidian usually possess Prochloron cells that are acquired from their mother colony. The process of vertical transmission of the photosymbionts has been histologically described in several species harboring Prochloron in the common cloacal cavity, where Prochloron cells are attached to the hairy surface of the tunic of the larvae during or just before larval spawning. Since the process has never been described in species harboring the photosymbionts in the tunic of the colony, here we describe the histological and ultrastructural process of vertical transmission in a photosymbiotic didemnid, Trididemnum miniatum. No photosymbionts were associated with gametes of the ascidian. Prochloron cells appeared in the epithelial pouch enclosing the embryo and were then incorporated in the tunic of embryos in late embryonic stages. Although Prochloron cells in the tunic of the colony were rarely associated with the host cells, some amoeboid-shaped tunic cells containing Prochloron cells were occasionally found around the epithelial pouch. Therefore, the host cell is thought to be a vehicle transporting the photosymbionts in the tunic of the colony to the tunic of the embryos. The photosymbiont cells were directly embedded in the tunic of the larval trunk. They were not contained in the host cells, as are those in the tunic of the colony. These observations revealed that the mechanism of vertical transmission in T. miniatum is very different from that in didemnids harboring Prochloron in the common cloacal cavity of the colonies, such as Trididemnum cyclops. The mechanisms of symbiont transmission are diverse even within the genus Trididemnum.     

Cellular localisation of secondary metabolites isolated from the Caribbean sponge <Emphasis Type="Italic">Plakortis simplex</Emphasis>

The Caribbean sponge, Plakortis simplex, is known to contain a large array of secondary metabolites, including the antimalarial polyketide plakortin, several unusual glycolipids, and some hopanoids, which closely resemble typical bacterial metabolites. The hypothesis that they could be products of bacterial metabolism was tested by localizing specific metabolites in cells using physical separation of sponge cells, bacterial symbionts and supernatant by differential centrifugation. The obtained fractions were analysed separately for the typical P. simplex metabolites by NMR and mass spectrometry, and most of them were shown to be present in the bacterial cells but not in the sponge cells. In addition, PCR screening showed that the biosynthetic pathway for glycosphingolipids was present in the bacterial cells. Isolation of a Sphingomonas strain PS193 from P. simplex and subsequent glycosphingolipid analysis resulted in the detection of a known glycosphingolipid, GSL-1, that did, however, not match the glycosphingolipid profile of P. simplex. Therefore, it is unlikely that Sphingomonas strain PS193 is an abundant member of the microbial community associated with P. simplex. Other glycosphingolipid producing bacteria in P. simplex remain to be identified. In conclusion, this study provides experimental evidence that the glycolipids and hopanoids and possibly also the polyketide plakortin are produced by microbial symbionts rather than the sponge from which the metabolites were originally isolated.     

Antennal responses to floral scents in the butterflies <Emphasis Type="Italic">Inachis io</Emphasis>, <Emphasis Type="Italic">Aglais urticae</Emphasis> (Nymphalidae), and <Emphasis Type="Italic">Gonepteryx rhamni</Emphasis> (Pieridae)

Summary. To better understand the biological role of floral scents for butterflies, electrophysiological responses to floral scents were investigated using combined gas chromatography and electroantennographic detection (GC-EAD). The antennal responses of three butterfly species, Aglais urticae L. (Nymphalidae), Inachis io L. (Nymphalidae), and Gonepteryx rhamni L. (Pieridae) to floral scent compounds from both natural and synthetic mixtures were examined. Floral scents were collected from the butterfly nectar plants Cirsium arvense (L.) (Asteraceae), and Buddleja davidii Franchet cv. (Loganicaeae) with dynamic head-space methods on Tenax-GR and eluted with pentane. These eluates, composed of natural floral scent blends, represent an array of compounds in their natural state. In the GC-EAD analyses eleven compounds were identified from C. arvense with the benzenoid compound phenylacetaldehyde in highest abundance. Seventeen compounds were identified from B. davidii with the irregular terpene oxoisophorone in highest abundance. Thirty-nine synthetic floral scent compounds were mixed in pentane, in equal amounts; about 35 ng were allowed to reach the antennae. The butterflies showed antennal responses to most of the floral scent compounds from both natural and synthetic blends except to the highly volatile monoterpene alkenes. Certain benzenoid compounds such as phenylacetaldehyde, monoterpenes such as linalool, and irregular terpenes such as oxoisophorone, were emitted in relatively large amounts from C. arvense and B. davidii, and elicited the strongest antennal responses. These compounds also elicited strong antennal responses when present in the synthetic scent blends. Thus, the butterflies seem to have many and /or sensitive antennal receptors for these compounds, which points to their biological importance. Moreover, these compounds are exclusively of floral scent origin. For B. davidii, which depends highly on butterflies for pollination, the exclusive floral scent compounds emitted in high abundance could be the result of an adaptive pressure to attract butterflies. Received 2 Septemter 2001; accepted 9 September 2002.     

Lack of sequestration of host plant glucosinolates in <Emphasis Type="Italic">Pieris rapae</Emphasis> and <Emphasis Type="Italic">P. grarricae</Emphasis>

Summary. Sequestration of plant toxins in herbivores is often correlated with aposematic coloration and gregarious behaviour. Larvae of Pieris brassicae show these conspicuous morphological and behavioural characteristics and were thus suggested to sequester glucosinolates that are characteristic secondary metabolites of their host plants. P. rapaeare camouflaged and solitary, and are thus not expected to sequester. To test this hypothesis and to check the repeatabi-lity of a study that did report the presence of the glucosinolate sinigrin in P. brassicae, larvae were reared on three species of Brassicaceae (Sinapis alba, Brassica nigra and Barbarea stricta), and different leaf and insect samples were taken for glucosinolate analysis. The major host plant glucosinolates could only be found in traces or not at all in larval haemolymph, bled or starved larvae, faeces or pupae of both species or P. brassicae regurgitant. Haemolymph of both Pieris spp. was not rejected by the ant Myrmica rubra in dual-choice assays; the regurgitant of P. brassicae was rejected. This suggests the presence of compounds other than glucosinolates that might be sequestered in or produced by P. brassicae only. In faeces of both Pieris spp. a compound which yielded 4-hydroxybenzylcyanide (HBC) upon incubation with sulfatase was detected in high concentrations when larvae had been reared on S. alba. This compound may be derived from hydrolysis of sinalbin, the main glucosinolate of that plant. The unidentified HBC progenitor was apparently not sequestered in the two Pieris spp., and was not detected in faeces of larvae reared on B. nigra or B. stricta. Received 18 July 2002; accepted 11 September 2002.     

Viable algae released by the seastar <Emphasis Type="Italic">Dermasterias</Emphasis><Emphasis Type="Italic">imbricata</Emphasis> feeding on the symbiotic sea anemone <Emphasis Type="Italic">Anthopleura elegantissima</Emphasis>

Echinoderms are major predators of anemones in temperate ecosystems. The fate of two algae, zooxanthellae and zoochlorellae, after their host anemone (Anthopleura elegantissima Brandt) was consumed by the leather star Dermasterias imbricata Grube was determined in experiments conducted in July and August 2004. Productivity, photosynthetic pigments, and mitotic index (percent of cells dividing) were used as indicators of algal health; algae released after leather stars consumed their host were compared with algae freshly isolated from anemones. Two types of waste products contained algae: pellets resulting from extraoral digestion, and feces. Zooxanthellae and zoochlorellae isolated from these waste products were photosynthetic, although to different extents. For algae from feces and pellets, light-saturated photosynthetic rates (P _max) were 85 and 13%, respectively, of P _max of freshly isolated zooxanthellae; and were 20 and 46%, respectively, for zoochlorellae. The photosynthetic pigments and mitotic index (percent of dividing cells) were not altered by the feeding activities of the leather star. These results show that algae released by seastar predation on their hosts remain viable, and are hence available for establishing symbioses in A. elegantissima and other potential hosts.     

Mate preferences in <Emphasis Type="Italic">Drosophila</Emphasis> infected with <Emphasis Type="Italic">Wolbachia</Emphasis>?

The maternally inherited bacterium Wolbachia pipientis generates strong reproductive incompatibilities between uninfected females and infected males (cytoplasmic incompatibility), significantly reducing both female and male reproductive success. Such fitness costs are thought to place selective pressure on hosts to evolve pre-copulatory preferences for mating with compatible mates, thereby enabling them to avoid the reproductive incompatibilities associated with Wolbachia. Therefore, uninfected females are predicted to prefer mating with uninfected males, whereas infected males are predicted to prefer mating with infected females. Despite these predictions, previous investigations of pre-copulatory mate preferences in Wolbachia-manipulated Drosophila have not found evidence of female preference for uninfected or compatible males. However, none of these studies utilised a design where focal individuals are provided with a simple choice in a relatively non-competitive situation. We examined both female and male pre-copulatory mate preference based on mate infection status in Drosophila simulans and D. melanogaster using simple choice assays involving between 30–50 replicates per treatment. Although we found no evidence of female pre-copulatory mate preferences in either species, male D. simulans exhibited some preference for mating with females of the same infection status. However, this preference was not evident when we repeated the experiment to confirm this finding. Consequently, we conclude that neither male nor female D. melanogaster and D. simulans exhibit significant Wolbachia-associated pre-copulatory mate preferences.     

Foraging responses in the butterflies <Emphasis Type="Italic">Inachis io</Emphasis>, <Emphasis Type="Italic">Aglais urticae</Emphasis> (Nymphalidae), and <Emphasis Type="Italic">Gonepteryx rhamni</Emphasis> (Pieridae) to floral scents

Summary. For butterflies to be efficient foragers, they need to be able to recognize rewarding flowers. Flower signals such as colours and scents assist this recognition process. For plant species to attract and keep butterflies as pollinators, species-specific floral signals are crucial. The aim of this study is to investigate foraging responses to floral scents in three temperate butterfly species, Inachis io L. (Nymphalidae), Aglais urticae L. (Nymphalidae), and Gonepteryx rhamni L. (Pieridae), in behavioural choice bioassays. The butterflies were allowed to choose bet-ween flower models varying in scent and colour (mauve or green). Flowers or vegetative parts from the plants Centaurea scabiosa L. (Asteraceae), Cirsium arvense (L.) (Asteraceae), Knautia arvensis (L.) (Dipsacaceae), Buddleja davidii Franchet (Loganicaeae), Origanum vulgareL. (Lamiaceae), Achillea millefolium L. (Asteraceae), and Philadelphus coronarius L. (Hydrangiaceae) were used as scent sources. All visits to the models — those that included probing and those that did not — were counted, as was the duration of these behaviours. Both flower-naive and flower-experienced (conditioned to sugar-water rewards, the colour mauve, and specific floral scents) butterflies were tested for their preference for floral versus vegetative scents, and to floral scent versus colour. The butterflies were also tested for their ability to switch floral scent preferences in response to rewards. Flower-naive butterflies demonstrated a preference for the floral scent of the butterfly-favourable plants C. arvense and K. arvensis over the floral scent of the non-favourable plants Achillea millefolium (Asteraceae), and Philadelphus coronarius cv. (Hydrangiaceae). Most of the butterflies that were conditioned to floral scents of either C. arvense, K. arvensis, or B. davidii readily switched theirfloral scent preferences to the one most recently associated with reward, thus demonstrating that floral scent constancy is a result from learning. These findings suggest that these butterflies use floral scent as an important cue signal to initially identify and subsequently recognize and distinguish among rewarding plants. Received 2 September 2001; accepted 9 September 2002.     

Dynamics of pigment degradation by the copepodite stage of <Emphasis Type="Italic">Pseudodiaptomus</Emphasis><Emphasis Type="Italic">euryhalinus</Emphasis> feeding on <Emphasis Type="Italic">Tetraselmis</Emphasis><Emphasis Type="Italic">suecica</Emphasis>

Short-term (3 h) changes in concentration of chlorophylls and their derivatives in stage V Pseudodiaptomus euryhalinus and their fecal material were followed by HPLC during a 24 h experiment. Copepodites were fed with the prasinophyte Tetraselmis suecica. Intact chlorophyll a and b were found in animals and fecal material and had similar dynamics of accumulation over time. The extent of transformation of chlorophyll a and b to colorless compounds was different with chlorophyll a being more extensively degraded. Additionally, several chlorophyll derivatives (pheophytin and pyropheophytin-like pigments) were found. Pyropheophytin a was the most abundant followed by pheophytin b, pheophytin a, and pheophorbide a. Relative amounts of pheopigments were different in copepodites and fecal material, and pheophytin a, pheophorbide a, and pheophytin b concentrations were low and variable. The amount of ingested chlorophyll recovered as chlorophyll a and its derivatives in fecal and copepodite pools was generally low (<5%), with one exception occurring after 9 h, when it accounted for >70%. These data suggest individual pheopigments are produced at different rates and that chemical or enzymatic mechanisms in the gut of copepodites act on the two chlorophylls in different ways.     

<Emphasis Type="Italic">Ceroplastes albolineatus</Emphasis>, the first scale insect shown to sequester pyrrolizidine alkaloids from its host-plant <Emphasis Type="Italic">Pittocaulon praecox</Emphasis>

Summary. The occurrence of pyrrolizidine alkaloids (PAs) in Pittocaulon (ex Senecio) praecox (Asteraceae) a species endemic to Mexico was established. The aboveground plant organs contain the 1,2-saturated monoester 7-angeloyl-dihydroxyheliotridane together with a small proportion of its 9-angeloyl isomer as major alkaloid. The monoesters are accompanied by the macrocyclic otonecine derivative senkirkine. Roots contain only related macrocyclic PAs with senecionine, senkirkine and platyphylline as major components; monoesters are absent. The broom-like succulent stems of P. praecox are infested by the scale insect Ceroplastes albolineatus conspicuously visible by its huge wax cover. All life-history stages, i.e. females, eggs, first instar nymphs (crawlers) and the wax cover were found associated with PAs. The measured PA concentrations clearly indicate sequestration. The highest PA concentrations (mg / g dry weight) reached are: mature females, 0.44; eggs, 0.58; crawlers, 0.37; wax cover, 0.08. The host plant as well as in the infesting scale insect contain the PAs exclusively as free bases. As a phloem-feeder C. albolineatus must acquire the PAs with the ingested phloem sap. This appears plausible since in Senecio species PA are transmitted and circulated through the phloem path. It is suggested that PAs may protect particularly the crawlers as the most endangered stage in the life-cycle of the scale insect.     

Effects of ocean temperature on the southern range limits of two understory kelps, <Emphasis Type="Italic">Pterygophora californica</Emphasis> and <Emphasis Type="Italic">Eisenia arborea</Emphasis>, at multiple life-stages

Environmental factors have long been shown to influence species distributions, with range limits often resulting from environmental stressors exceeding organism tolerances. However, these abiotic factors may differentially affect species with multiple life-history stages. Between September 2004 and January 2006, the roles of temperature and nutrient availability in explaining the southern distributions of two understory kelps, Pterygophora californica and Eisenia arborea (Phaeophyceae, Laminariales), were investigated along the coast of California, USA and the Baja California Peninsula, Mexico, by limiting either: (a) tissue nitrogen uptake and storage by adult sporophytes during periods of elevated temperature, and/or (b) production of embryonic sporophytes by microscopic gametophytes. Results suggest that while adult sporophytes of both species are tolerant of high temperatures and low nutrients, reproduction by their microscopic stages is not. Specifically, while E. arborea produced embryonic sporophytes at both 12 and 18°C, temperatures commonly observed throughout the southern portion of its range, P. californica produced sporophytes at 12 but not at 18°C. As a result, it appears that the southern distribution of P. californica, which ends in northern Baja California, Mexico, may be limited by temperature acting on its microscopic stages. In contrast, the ability of E. arborea’s microscopic and adult stages to tolerate elevated temperatures allows it to persist in the warmer southern waters of Baja California, as well as to the north along the California coast where both species co-occur.     

Constitutive or induced defences - how does <Emphasis Type="Italic">Eucalyptus globulus</Emphasis> defend itself from larval feeding?

Summary. Following herbivory, induced responses involving plant secondary metabolites have been reported in a number of tree species. Although a wide range of plant secondary metabolites appear to operate as constitutive plant defences in trees belonging to the Eucalyptus genus, no induced responses have as yet been reported following foliar-chewing insect damage. We empirically tested whether branch defoliation (artificial and larval) of 2-year-old Eucalyptus globulus Labill. trees altered the abundance of specific plant secondary metabolites immediately (3 months after initial larval feeding) and 8 months after the cessation of larval feeding. Metabolites assayed, included essential oils, polyphenolic groups and foliar wax compounds and in all cases their abundance was not significantly altered by defoliation. However, the level of foliar tannins after 3 months of larval feeding did display a trend that suggested elevated levels as the result of defoliation, though this trend was not evident 8 months later, indicating that, if real, the response was a rapid and not a delayed induced response. The level of foliar tannins was also negatively correlated to both average larval survival and average percentage branch defoliation, suggesting that foliar tannins may operate as toxins and/or anti-feedants to M. privata larval feeding.     

Inter-polyp genetic and physiological characterisation of <Emphasis Type="Italic">Symbiodinium</Emphasis> in an <Emphasis Type="Italic">Acropora valida</Emphasis> colony

Corals harbouring genetically mixed communities of endosymbiotic algae (Symbiodinium) often show distribution patterns in accordance with differences in light climate across an individual colony. However, the physiology of these genetically characterised communities is not well understood. Single stranded conformation polymorphism (SSCP) and real time quantitative polymerase chain reaction (qPCR) analyses were used to examine the genetic diversity of the Symbiodinium community in hospite across an individual colony of Acropora valida at the spatial scale of single polyps. The physiological characteristics of the polyps were examined prior to sampling with a combined O₂ microelectrode with a fibre-optic microprobe (combined sensor diameter 50–100 μm) enabling simultaneous measurements of O₂ concentration, gross photosynthesis rate and photosystem II (PSII) quantum yield at the coral surface as a function of increasing irradiances. Both sun- and shade-adapted polyps were found to harbour either Symbiodinium clade C types alone or clades A and C simultaneously. Polyps were grouped in two categories according to (1) their orientation towardps light, or (2) their symbiont community composition. Physiological differences were not detected between sun- and shade-adapted polyps, but O₂ concentration at 1,100 μmol photons m⁻² s⁻¹ was higher in polyps that harboured both clades A and C symbionts than in polyps that harboured clade C only. These results suggest that the acclimatisation of zooxanthellae of individual polyps of an A. valida colony to ambient light levels may not be the only determinant of the photosynthetic capacity of zooxanthellae. Here, we found that photosynthetic capacity is also likely to have a strong genetic basis and differs between genetically distinct Symbiodinium types.     

Cloning and expression of a tauropine dehydrogenase from the marine sponge <Emphasis Type="Italic">Suberites </Emphasis><Emphasis Type="Italic">domuncula</Emphasis>

The cDNA sequence coding for tauropine dehydrogenase (TaDH) [belonging to the family of opine dehydrogenases] has been determined. Using the demosponge Suberites domuncula, we describe for the first time the tauropine dehydrogenase gene (of length 2,992 kb) from a eukaryote, consisting of two introns flanked by three exons. Moreover, two allelic variants have been identified, which are present in the different specimens either in a homozygotic or in a heterozygotic way; the data suggest an intermediary type of heritance. Phylogenetic analyses indicate that S. domuncula TaDH is only distantly related to the opine dehydrogenases from marine invertebrates; rather it comprises high sequence similarity to bacterial ornithine cyclodeaminases (OCD). In addition, expression studies revealed that the steady-state level of TaDH dropped drastically in animals, which had been exposed to elevated aeration. Antibodies raised against the recombinant sponge TaDH were used to demonstrate that S. domuncula expresses high levels of this enzyme in almost all cells. If tissue samples were kept under additional aeration no immuno-signals could be identified. A strong accumulation of the enzyme was seen around the bacteria, existing in bacteriocytes, indicating that under aerobic conditions the bacteria might produce taurine. These data suggest involvement of the sponge TaDH in the final step of the glycolytic pathway, more specifically, in regeneration of NAD(+) under anaerobic conditions. Furthermore, potential mutual influences between bacteria and sponge are discussed, claiming a horizontal gene transfer of the gene from a bacterium to the sponge. The following sequences from Suberites domuncula have been deposited (EMBL/GenBank): the cDNA encoding the tauropine [(carboxyethyl)-taurine/derived from sulfhydryl-amino acids] dehydrogenase (SD_TaDH) under AM712888; the corresponding gene under AM712889; and the fragments of the sponge-associated bacteria comprising the opine dehydrogenase domain: SUBDO_BAC1 (AM712890), SUBDO_BAC2 (AM712891), SUBDO_BAC3 (AM712892), SUBDO_BAC5 (AM712894), SUBDO_BAC6 (AM712895), SUBDO_BAC7 (AM712896), SUBDO_BAC8 (AM712897) and SUBDO_BAC9 (AM712898).     

On the consortium of the tintinnid <Emphasis Type="Italic">Eutintinnus</Emphasis> and the diatom <Emphasis Type="Italic">Chaetoceros</Emphasis> in the Pacific Ocean

The morphology and distribution of the diatoms Chaetoceros tetrastichon and Ch. dadayi as epiphytes on the loricae of the tintinnids Eutintinnus apertus and E. pinguis investigated in the open waters of the Pacific Ocean. The Eutintinnus–Chaetoceros consortia was encountered in 38 of the 52 sampling stations from 34°N to 33°S, and together were among represented the most wide-spread species. The abundance was low with a maximum of 32 consortia l⁻¹ and E. apertus was often the most abundant species of the genus. The free-living Eutintinnus congeneric species showed a wider vertical distribution, whereas E. apertus–Chaetoceros tended to be near the surface. The success of E. apertus in consortium with Chaetoceros may be due to increase of the clearance rate and/or the lower susceptibility to predation. Chaetoceros modifies its morphology to adapt the epiphytic life, especially Ch. dadayi. The shorter curved setae may facilitate the transfer to the lorica of the daughter tintinnid after the cell division. The free-living Ch. tetrastichon and Ch. dadayi are very rare and Chaetoceros remained attached to empty loricae or encysted tintinnid cells. This suggests that the Eutintinnus–Chaetoceros consortium is obligate for the success of the diatom and renders the tintinnid more competitive versus congeneric species.     

Intraguild predatory interactions between the jellyfish <Emphasis Type="Italic">Cyanea capillata</Emphasis> and <Emphasis Type="Italic">Aurelia aurita</Emphasis>

While qualitative observations of jellyfish intraguild predation abound in the literature, there are only few rate measurements of these interactions. We quantified predation rates among two common jellyfish in northern boreal waters, Cyanea capillata and its prey Aurelia aurita, both of which also feed on crustacean zooplankton and fish larvae. A series of incubation experiments using a wide range of prey concentrations (0.38–3.8 m⁻³) in large containers (2.6 m³) was carried out. By replenishing the prey continuously as they were captured we maintained a nearly constant prey concentrations. Ingestion rates increased linearly up to prey concentrations of 1.92 m⁻³, yielding maximum clearance rates of ∼2.37 ± 0.39 m³ predator⁻¹ h⁻¹ for C. capillata predators 16 ± 2.3 cm in diameter. Mean ingestion rate at saturated prey concentrations (1.92–3.85 m⁻³) was 4.01 ± 0.78 prey predator⁻¹ h⁻¹. Behavioral observations suggested that predators did not alter their swimming behavior during meals, and thus that feeding rates were generally handling limited rather than encounter limited. Predators captured more prey than needed, and semi-digested prey was often discarded when fresh prey was encountered.     

How <Emphasis Type="Italic">Doratomyces stemonitis</Emphasis> copes with Benzoxazolin-2(3<Emphasis Type="Italic">H</Emphasis>)-one (BOA), its derivatives and detoxification products

Summary. Doratomyces stemonitis (Hyphomycetales, Dematiaceae) is a saprotrophic fungus belonging to the mycobiota of the cereal rhizosphere. The fungus is able to metabolize benzoxazolin-2-(3H)-one and a variety of its derivatives including higher plant detoxification products, microbial degradation products and the chemically rather stable 2-amino-(3H)-phenoxazin-3-one. D. stemonitis can use all of these compounds as sole C-sources but their utilization, especially that of microbial degradation products and 2-amino-(3H)-phenoxazin-3-one, seems to be highly energy consuming, resulting in slow mycelium growth and a change of colony morphology. Benzoxazolin-2-(3H)-one derived compounds induce the synthesis of different isoforms of a glycosylated protein with sequence homologies to the endo-1,3-β-glucanase Asp f2, an allergen from Aspergillus fumigatus and other Asp f2-like proteins e.g., from Verticillium dahliae or PRA1 antigen from Candida albicans. The induction of the protein is regarded as a stress response.     

Monitoring acoustically tagged king prawns <Emphasis Type="Italic">Penaeus</Emphasis> (<Emphasis Type="Italic">Melicertus</Emphasis>) <Emphasis Type="Italic">plebejus</Emphasis> in an estuarine lagoon

Fine-scale movement patterns in penaeid prawns are rarely observed in situ, but are essential in understanding habitat use, foraging, and anti-predator behaviour. Acoustic telemetry was applied to examine the activity, space utilization, and habitat use of the eastern king prawn Penaeus (Melicertus) plebejus, at small temporal and spatial scales. Tracking of sub-adult P. plebejus (n = 9) in Wallagoot Lake (36.789°S, 149.959°E; 23 April–12 May 2009) and calculation of a minimum activity index (MAI) revealed high variation in activity rates across diel periods and in different habitats. Elevated activity rates and movement indicated foraging in unvegetated habitats during the night. Areas within the 95 and 50% space utilization contours averaged 2,654.1 ± 502.0 and 379.9 ± 103.9 m², respectively, and there was a significant negative relationship between these areas and prawn activity rates in unvegetated habitats. This study provides the first estimates of prawn activity rates and space utilization in the field. Application of acoustic telemetry can increase knowledge of prawn movements and their interactions with other marine species in different habitats.     

Degree-day accumulation controlling allopatric and sympatric variations in the sociality of sweat bees, <Emphasis Type="Italic">Lasioglossum</Emphasis> (<Emphasis Type="Italic">Evylaeus</Emphasis>) <Emphasis Type="Italic">baleicum</Emphasis> (Hymenoptera: Halictidae)

Halictine bees exhibit a wide range of social behaviour that varies both inter- and intraspecifically. Although previous studies suggested that the intraspecific variation might be attributed to temperature differences, there was no direct evidence to detect the relationships between temperature and socialities. Lasioglossum (Evylaeus) baleicum exhibited solitary behaviour in a cooler locality (Kawakita) because of the shorter breeding season; in a warmer locality (Nishioka Park), however, this bee species exhibited eusociality at sunny site and solitary behaviour at shady site, whereas a molecular phylogeny confirms that all of these colonies are evidently conspecific. Therefore, we examined the effect of degree-day accumulation on the sympatric social variation of L. baleicum by rearing the bees to calculate the threshold temperature. Whereas they showed high mortality, the threshold temperature was estimated to be 10.33°C and the expected degree-day accumulation was 340 degree days. When we use this value of a degree-day accumulation to estimate the expected eclosion date, the estimated dates were always consistent with observed eclosion dates. In any sites where the bees were solitary, the degree-day accumulation was not enough for the second eclosion by the end of the bee-active season. In Nishioka Park, sex ratio of the first brood was female biased, and daughters were smaller than mothers; in Kawakita, however, there was no sex bias, and daughters were as large as their mothers indicating that the foundresses seem to produce gyne-sized females in Kawakita but worker-sized females in Nishioka though these females do not become workers at shady site.     

Sugar compound products of the periphytic diatom <Emphasis Type="Italic">Navicula ramosissima</Emphasis> induce larval settlement in the barnacle, <Emphasis Type="Italic">Amphibalanus amphitrite</Emphasis>

This study investigated the response of cyprids of the barnacle Amphibalanus amphitrite to 23 strains of laboratory cultured periphytic diatoms isolated from microbial biofilms that formed on glass slides immersed in Tachibana bay, Nagasaki, and those from mass-production tanks in the Fisheries Center of Nagasaki City, Japan. In addition, periphytic diatoms were subjected to various treatments, in order to investigate the nature of the chemical cue in periphytic diatoms. Cyprids of A. amphitrite responded differently to the 23 different periphytic diatom strains and settled in high percentages on Cocconeis sp. and Navicula ramosissima strain A. On the other hand, nine strains of diatoms significantly inhibited settlement. The settlement inducing activity of N. ramosissima strain A increased linearly with diatom density, and its activity was enhanced by culturing the diatom under a bacteria-free condition, suggesting that specific diatom species, i.e., N. ramosissima strain A, may play an important role on larval settlement of the barnacle. Subjecting N. ramosissima strain A biofilm to hydrochloric acid (HCl) and ethanol (EtOH) treatments or heating it at 100°C did not inactivate the film, indicating that the settlement cue was a stable surface bound compound that did not decompose from the above treatments. Moreover, of the various lectins, enzymes, and drugs [H₅IO₆ and sodium dodecyl sulfate or (SDS)] used, only Lentil Agglutinin (LCA) treatment of N. ramosissima strain A biofilm resulted in the reduction its settlement inducing activity. A positive correlation was observed between the settlement inducing activity and the amount of LCA conjugated fluorescein isothiocyanate (FITC-LCA) of N. ramosissima strain A. On the other hand, subjecting biofilms of N. ramosissima strain B, an inactive strain, to various types of treatments resulted in the induction of A. amphitrite larval settlement but LCA treatment also reduced the activity of these treated N. ramosissima strain B biofilms. These findings suggest that a cue containing an LCA-binding sugar chain is present in both A and B strains of N. ramosissima but the large amount of mucous substance covering N. ramosissima strain B biofilm probably makes the sugar chain containing active subunit in strain B unavailable to A. amphitrite cyprids. In conclusion, periphytic diatoms such as N. ramosissima play an important role in larval settlement of the barnacle A. amphitrite. The cue in the diatom was an LCA-binding sugar chain(s) compound that may have similarities to the settlement inducing protein complex (SIPC) from adult shell of the barnacle.     

Evidence of a spawning area of <Emphasis Type="Italic">Anguilla</Emphasis><Emphasis Type="Italic">marmorata</Emphasis> in the western North Pacific

Leptocephali of the tropical eel Anguilla marmorata have been consistently collected in the same area of the North Equatorial Current (NEC) in the western North Pacific during three consecutive cruises in June and July of 1991 (N=28) and 1994 (N=20), and July and September of 1995 (N=27), indicating that this is a spawning area of this species. These leptocephali were collected from 130°E to 142°E and 12°N to 20°N, to the west of the Mariana Islands, in 20 tows in 1991, in 13 tows in 1994 and in 17 tows in 1995, indicating a widespread presence, but a relatively low abundance. Six of these specimens (16.3-36.0 mm total length) from the 1995 cruise, which were of the typical size range of these leptocephali, were genetically confirmed to be A. marmorata in a previous study. The consistent presence of recently spawned A. marmorata leptocephali (9-20 mm) in all 3 years, suggests that the western region of the NEC is the spawning area of the northern population of A. marmorata that was identified in a recent population genetics study. These leptocephali would thus be transported westward by the NEC and then transported north into the Kuroshio Current and toward Taiwan and Japan, or south toward the southern Philippines and into the Celebes Sea by the Mindanao Current. Available evidence indicates that A. marmorata may have potentially year-round spawning, and the presence of a spawning area of this species in the same region as that of Anguilla japonica suggests that the northern population of A. marmorata has evolved a spawning migration from East Asia, the Philippines and the Celebes Sea region to the NEC area, but differs from A. japonica by having some individuals that recruit to more southern areas.