Okadaic acid: a potential defense molecule for the sponge <Emphasis Type="Italic">Suberites domuncula</Emphasis>

Sponges are sessile filter feeders that are exposed to toxic compounds/xenobiotics and a huge number of diverse pro- and eukaryotic organisms occurring in the aqueous milieu. To cope with these threats, sponges have developed a variety of chemical and immunological defense systems. Among them are potent secondary metabolites or toxins. The demosponge Suberites domuncula contains the bioactive compound okadaic acid; concentrations of ᅢ ng/g (ᅬ nM) of wet tissue have been quantified by LC/MS. In the present study, we sought to determine whether okadaic acid is beneficial for this sponge. Antibodies were raised against okadaic acid and used to localize this compound in situ. It could be demonstrated that the polyclonal antibodies/antiserum stained the sponge cells (endopinacocytes) which form the epithelium of the lacunae and the water channels. They also reacted with bacteria present in sponge tissue. Okadaic acid was found to augment the immune response of S. domuncula against the bacterial endotoxin lipopolysaccharide (LPS). If primmorphs, a special form of 3-D aggregates, are incubated with LPS at a concentration of 3 µg/ml, a reduction in the incorporation rate of [³H] phenylalanine is measured. In the presence of okadaic acid, this effect is measured already at a concentration of 1 µg/ml, suggesting that okadaic acid sensitizes this (immune) response. Okadaic acid is a known inhibitor of protein phosphatases which, as shown here, cause an increased phosphorylation of p38, a central kinase of the MAP kinase pathway; this may explain the sensitive response of the sponge cells to LPS. At higher concentrations, okadaic acid induces apoptosis, as was demonstrated here by a differential expression of the Bcl-2 homolog and the caspase gene. It is concluded that okadaic acid has two functions in the sponge; first, at low concentrations (<100 nM) a stimulation of the defense system against bacteria, and second, at concentrations above 500 nM, a differential effect on the pro-/anti-apoptotic genes.     

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.     

Spatial distribution patterns of the gorgonians <Emphasis Type=Italic>Eunicella singularis</Emphasis>, <Emphasis Type=Italic>Paramuricea clavata</Emphasis>, and <Emphasis Type=Italic>Leptogorgia sarmentosa</Emphasis> (Cape of Creus,Northwestern Mediterranean Sea)

Most of the current knowledge on Mediterranean gorgonians is restricted to investigations of those populations found within shallow sublittoral waters, and only limited data are available for populations located below scuba depth. To overcome this lack of information, the occurrence and abundance of the gorgonians Eunicella singularis, Paramuricea clavata, and Leptogorgia sarmentosa were investigated in northwestern Mediterranean benthic communities over a wide geographical (60 km of coastline) and bathymetrical (0–70 m deep) extent using a remotely operated vehicle (ROV). The greatest occurrence and abundances of E. singularis and P. clavata were concentrated in areas that are directly exposed to strong near-bottom currents. E. singularis was the most common and abundant species and displayed great plasticity and amplitude in its environmental preferences. Conversely, P. clavata showed a very patchy distribution that was associated with vertical rocky walls. Only isolated colonies of L. sarmentosa were observed in the study area. Hot spots of abundance of E. singularis and P. clavata were identified below a depth of 40 m, which demonstrates the importance of studying the distribution of benthic species over a wide geographical and bathymetrical extent.     

Interactions between an invasive and a native bryozoan (<Emphasis Type=Italic>Membranipora membranacea</Emphasis> and <Emphasis Type=Italic>Electra pilosa</Emphasis>) species on kelp and <Emphasis Type=Italic>Fucus</Emphasis> substrates in Nova Scotia,Canada

Most research on biological invasions to date has focused on the population dynamics of very successful and disruptive introduced species; however, additional knowledge of the biology of the native species is essential for understanding interactions between the two and may reveal factors that limit invasion success. The invasive bryozoan Membranipora membranacea interacts with native Electra pilosa on two substrates in northwest Atlantic subtidal habitats: highly dynamic and fast-growing kelps; and smaller, more stable, and slow-growing fucoid algae. We quantified the relative abundance and evaluated encounter outcomes in different seasons of these two bryozoans on kelp and Fucus at four sites in Nova Scotia. We also examined the effects of substrate (kelp, Fucus), temperature (7, 10, 13°C), and food (limited, unlimited) on growth rates of E. pilosa in laboratory experiments and using field manipulations. We compared our findings on factors affecting the growth of E. pilosa to those on M. membranacea obtained in similar and thus directly comparable experiments from a previous study. The proportional abundance of M. membranacea was greater than that of E. pilosa on kelps, but the opposite was observed on Fucus. Competitive standoffs between the two bryozoans were more frequent than expected, with no differences recorded between substrates; most encounters were won by M. membranacea. Growth of E. pilosa was faster on Fucus than kelp, decreased with increasing temperature only on Fucus, and was not affected by food. Growth rate of E. pilosa in all treatments was slower than that previously measured for M. membranacea. Faster growth and strong overgrowth abilities likely interact on kelps to ensure success of the invasive bryozoan. Success can be limited by low space availability, which in turn restricts growth rate, and consequently, colony size, such as on fucoid substrates. The incorporation of alternative contexts into invasion research can reveal factors involved in the resilience of native communities.     

Sympatry in grapsoid crabs (genera <Emphasis Type=Italic>Planes</Emphasis> and <Emphasis Type=Italic>Plagusia</Emphasis>) from olive ridley sea turtles (<Emphasis Type=Italic>Lepidochelys olivacea</Emphasis>), with descriptions of crab diets and masticatory structures

Grapsoid crabs of the genera Planes and Plagusia are commonly referred to as “rafting crabs” due to their propensity to live on flotsam and pelagic marine animals. Planes minutus and Planes major (=Planes cyaneus) are epibionts of sea turtles. Occurrences of grapsoid crabs in the genera Planes and Plagusia were evaluated on a total of 27 olive ridley sea turtles, Lepidochelys olivacea, from the eastern tropic Pacific (1998–2001) and the Hawaiian Islands (2002) captured in July–December each year. This is the first report of Planes marinus and Plagusia squamosa on sea turtles, and of P. major, P. marinus, and P. squamosa in sympatry on a confined substrate. Stomach content analyses showed P. major and P. marinus consumed a variety of neuston and marine vegetation, with the former consuming considerably more animal material. Epibiotic P. squamosa consumed mostly plant material. The three Planes species had distinctive differences in gastric mill tooth morphology. The versatile mouthparts of P. marinus are described and resemble those of their congeners. Most female P. major and P. marinus collected were ovigerous and present in all survey months.     

Molecular characterization of the zoanthid genus <Emphasis Type=Italic>Isaurus</Emphasis> (Anthozoa: Hexacorallia) and associated zooxanthellae (<Emphasis Type=Italic>Symbiodinium</Emphasis> spp.) from Japan

The zoanthid genus Isaurus (Anthozoa: Hexacorallia) is known from both the Indo-Pacific and Atlantic Oceans, but phylogenetic studies examining Isaurus using molecular markers have not yet been conducted. Here, two genes of markers [mitochondrial cytochrome oxidase subunit I (COI) and mitochondrial 16S ribosomal DNA (mt 16S rDNA)] from Isaurus specimens collected from southern Japan (n = 19) and western Australia (n = 3) were sequenced in order to investigate the molecular phylogenetic position of Isaurus within the order Zoantharia and the family Zoanthidae. Additionally, obtained sequences and morphological data (polyp size, mesentery numbers, mesogleal thickness) were utilized to examine Isaurus species diversity and morphological variation. By comparing our obtained sequences with the few previously acquired sequences of genera Isaurus as well as with Zoanthus, Acrozoanthus (both family Zoanthidae), and Palythoa spp. (family Spenophidae) sequences, the phylogenetic position of Isaurus as sister to Zoanthus within the Family Zoanthidae was suggested. Based on genetic data, Isaurus is most closely related to the genus Zoanthus. Despite considerable morphological variation (in particular, polyp length, mesentery numbers, external coloration) between collected Isaurus specimens, all specimens examined are apparently conspecific or very closely related based on molecular data and observed morphological variation within colonies. Additionally, obtained internal transcribed spacer of ribosomal DNA (ITS-rDNA) sequences from symbiotic zooxanthellae (Symbiodinium spp.) from all Isaurus specimens were shown to be subclade C1-related Symbiodinium. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Size-dependent photosynthetic performance in the giant clam <Emphasis Type=Italic>Tridacna maxima</Emphasis>, a mixotrophic marine bivalve

Giant clams form a symbiosis with photosynthetic algae of the genus Symbiodinium that reside in clam mantle tissue. The allometry of symbiont photosynthetic performance was investigated as a mechanism for the increasing percentage of giant clam carbon respiratory requirements provided by symbionts as clam size increases. Chlorophyll fluorescence measurements of symbionts of the giant clam Tridacna maxima were measured during experiments conducted in September of 2009 using specimens 0.5–200 g tissue wet weight (3–25 cm long), collected from waters around southern Taiwan (N 21°36′, E 120°47′) from July to August of 2009. Light-dependent decreases in effective quantum yield (∆F/F _m′) calculated as the noontime maximum excitation pressure over PSII (Q _m), relative electron transport rates (rETR), and dark-adapted maximum quantum yield (F _v/F _m) all varied as a quadratic function of clam size. Both Q _m and rETR increased as clam size increased up to ~10–50 g then decreased as clam size increased. F _v/F _m decreased as clam size increased up to ~5–50 g then increased as clam size increased. Chlorophyll fluorescence measurements of rETR were positively correlated with gross primary production measured during chamber incubations. Overall, symbionts of mid-sized clams ~5–50 g exhibited the highest light-dependent decreases in effective photosynthetic efficiencies, the highest relative electron transport rates, and the lowest maximum photosynthetic efficiencies, and symbiont photosynthetic performance is allometric with respect to host clam size.     

The invasive venomous striped eel catfish <Emphasis Type=Italic>Plotosus</Emphasis><Emphasis Type=Italic>lineatus</Emphasis> in the Levant: possible mechanisms facilitating its rapid invasional success

The venomous striped eel catfish Plotosus lineatus was first recorded in the Mediterranean in 2002. Within 1–3 years, it has spread throughout the entire Israeli coast. We have studied its spatiotemporal distribution patterns via trawl surveys in order to determine the scale and extent of this invasion. Findings indicate that a population explosion has occurred, and the catfish now inhabits all sandy and muddy substrates up to ca 80 m. P. lineatus was found to recruit in autumn in the Mediterranean and displays similar or improved growth patterns and condition factor compared to those found in its native habitat. We discuss the possible ecological mechanisms responsible for its success: Benthic invaders are among its main prey items, suggesting an invasional meltdown process. We also point to the decline of indigenous species using its trophic and behavioral–ecological niche and hypothesize that they might be outcompeted and displaced by the catfish.     

Comparison of the statolith structures of <Emphasis Type=Italic>Chironex fleckeri</Emphasis> (Cnidaria,Cubozoa) and <Emphasis Type=Italic>Periphylla periphylla</Emphasis> (Cnidaria,Scyphozoa): a phylogenetic approach

The rhopalia and statocysts of Periphylla periphylla (Péron and Lesueur in Ann Mus Hist Nat Marseille 14:316–366,1809) and Chironex fleckeri Southcott (Aust J Mar Freshw Res 7(2):254–280 1956) were examined histologically and showed several homologous characteristics. Differences in sensory area distribution could be connected to a slightly different functionality of equilibrium sensing. In P. periphylla, the statoliths (crystals) grow independently of each other; whereas in C. fleckeri, one large crystal covers the smaller ones. The structures of both statoliths were examined in detail with single-crystal diffraction, microtomography and diffraction contrast tomography. The single compact statolith of C. fleckeri consisted of bassanite as was previously known only for other rhopaliophoran medusae. An origin area with several small oligocrystals was located in the centre of the cubozoan statolith. The origin areas and the accretion of statoliths are similar in both species. Our results lead to the assumption that the single bassanite statolith of C. fleckeri (Cnidaria, Cubozoa) is a progression of the scyphozoan multiplex statolith. It is therefore suggested that the Cubozoa are derived from a scyphozoan ancestor and are a highly developed taxa within the Rhopaliophora.     

Culturable actinobacteria isolated from marine sponge <Emphasis Type="Italic">Iotrochota</Emphasis> sp.

The study describes the diversity of actinobacteria isolated from the marine sponge Iotrochota sp. collected in the South China Sea. Species and natural product diversity of isolates were analyzed, including screening for genes encoding polyketide synthases (PKS) and nonribosomal peptide synthetase (NRPS), and 16S rRNA gene restriction fragment length polymorphism (RFLP). PKS and NRPS sequences were detected in more than half of the isolates and the different “PKS-I–PKS-II–NRPS” combinations in different isolates belonging to the same species indicated a potential natural product diversity and divergent genetic evolution. The phylogenetic analysis based on 16S rRNA gene sequencing showed that the isolates belonged to genera Streptomyces, Cellulosimicrobium, and Nocardiopsis. The majority of the strains tested belonged to the genus Streptomyces and one of them may be a new species. To our knowledge, this is the first report of a bacterium classified as Cellulosimicrobium sp. isolated from a marine sponge. Key Laboratory of Marine Bio-recourses Sustainable Utilization (LMB-CAS), Guangdong Key Laboratory of Marine Materia Medica (LMMM-GD), South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, People’s Republic of China.     

Superoxide dismutase in the marine sponge <Emphasis Type="Italic">Cliona celata</Emphasis>

The aim of this work is to investigate the activity of the antioxidant enzyme superoxide dismutase in the cosmopolitan sponge Cliona celata (Grant, 1826), since this enzyme has been described as a useful biomarker for marine pollution in other marine invertebrates. The quantification of the catalytic activity for superoxide dismutase is quite complex because its substrate is an unstable free radical. Several methods have been developed for this enzymatic activity determination; most of them are based on inhibition of a redox reaction involving the superoxide radical. In this work, two methods are compared, for crude sponge extracts, as far as repeatability, reproducibility and sensibility are concerned. The adrenaline oxidation method seems to be the most adequate for these determinations. Statistical treatment of the data indicates that the reference value for the specific superoxide dismutase activity in C. celata should be in the interval [0–535.5] U/mg of total protein in wet tissues, for normal populations.     

Gene expression profiles of the swimming crab <Emphasis Type=Italic>Portunus trituberculatus</Emphasis> exposed to salinity stress

The swimming crab, Portunus trituberculatus, is an important marine fishery and aquaculture species. Although P. trituberculatus is a euryhaline species, water salinity condition influenced its distribution, migration route, and artificial propagations. To investigate gene expression in the P. trituberculatus exposed to different salinity stresses, 2426 expressed sequence tags (ESTs) from gill cDNA library were selected to spot on a cDNA microarray chip. In total, 417 differentially expressed genes were identified and grouped into eight clusters by hierarchical clustering analysis. Approximately 71.5% of grouped genes belonged to three independent expression patterns, indicating that these three expression patterns may represent three important stress tolerance pathways or networks in P. trituberculatus. Moreover, our cDNA microarray data suggested that there were differences in gene expression patterns of P. trituberculatus for low salinity and high salinity acclimation, suggesting that two salinity challenges resulted in a wide variation of gene expression in P. trituberculatus. In addition, a series of genes such as CCAAT/enhancer-binding protein, Na/K ATPase β-subunit, and heat shock proteins (HSPs) genes were suggested to be key elements during salinity acclimation process. Overall, this work represented an important step toward understanding the molecular processes and mechanisms involved in salinity acclimation of the swimming crab.     

Fine-scale observations of the predatory behaviour of the carnivorous copepod <Emphasis Type=Italic>Paraeuchaeta norvegica</Emphasis> and the escape responses of their ichthyoplankton prey,Atlantic cod (<Emphasis Type=Italic>Gadus morhua</Emphasis>)

Paraeuchaeta norvegica (8.5 mm total length) and yolk-sac stage Atlantic cod larvae (4 mm total length) (Gadus morhua) larvae were observed in aquaria (3 l of water) using silhouette video photography. This allowed direct observations (and quantitative measurement) of predator–prey interactions between these two species in 3-dimensions. Tail beats, used by cod larvae to propel themselves through the viscous fluid environment, also generate signals detectable by mechanoreceptive copepod predators. When the prey is close enough for detection and successful capture (approximately half a body-length), the copepod launches an extremely rapid high Reynolds number attack, grabbing the larva around its midsection. While capture itself takes place in milliseconds, minutes are required to subdue and completely ingest a cod larva. The behavioural observations are used to estimate the hydrodynamic signal strength of the cod larva’s tail beats and the copepod’s perceptive field for larval fish prey. Cod larvae are more sensitive to fluid velocity than P. norvegica and also appear capable of distinguishing between the signal generated by a swimming and an attacking copepod. However, the copepod can lunge at much faster velocities than a yolk-sac cod larva can escape, leading to the larva’s capture. These observations can serve as input to the predator–prey component of ecosystem models intended to assess the impact of P. norvegica on cod larvae.     

Mechanism(s) of long chain n-3 essential fatty acid production in two species of heterotrophic protists: <Emphasis Type=Italic>Oxyrrhis marina</Emphasis> and <Emphasis Type=Italic>Gyrodinium dominans</Emphasis>

As intermediaries, some heterotrophic protists can enhance the content of the long chain n-3 essential fatty acids (LCn-3EFAs), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), of low food quality algae for subsequent use at higher trophic levels. However, the mechanisms that produce LCn-3EFAs are presently unknown, although LCn-3EFA production by heterotrophic protists at the phytoplankton–zooplankton interface may potentially affect the nutritional status of the pelagic system. We investigated whether the heterotrophic protists, Oxyrrhis marina and Gyrodinium dominans, produce LCn-3EFAs via elongation and desaturation of dietary LCn-3EFA precursors and/or synthesize LCn-3EFAs de novo by: (1) feeding the two heterotrophic protists with a prey deficient in n-3 fatty acids, (2) incubating them in medium containing ¹³C-labeled sodium acetate, and (3) feeding the two protists gelatin acacia microspheres (GAMs) containing a deuterium-labeled LCn-3EFA precursor, linolenic acid [18:3(n-3)-d4]. Both O. marina and G. dominans synthesized EPA and DHA when fed the n-3 fatty acid-deficient prey, Perkinsus marinus, a parasitic protozoan. O. marina, but not G. dominans utilized ¹³C-labeled acetate from the medium to produce uniformly labeled fatty acids, including DHA. Both heterotroph species consumed GAMs containing 18:3(n-3)-d4 and catabolized 18:3(n-3)-d4 to 16:3(n-3)-d4 and 14:3(n-3)-d4, while no 20 or 22 carbon metabolites of 18:3(n-3)-d4 were detected. These results suggest that O. marina and G. dominans do not elongate and desaturate dietary LCn-3EFA precursors to produce LCn-3EFAs, but rather they produce LCn-3EFAs de novo, possibly via a polyketide synthesis pathway.     

Aerial surveys of manatees (<Emphasis Type=Italic>Trichechus manatus)</Emphasis> in Lee County,Florida<Emphasis Type=Italic>,</Emphasis> provide insights regarding manatee abundance and real time information for managers and enforcement officers

Conservation and management of the endangered Florida manatee is often centered on reducing mortality caused by watercraft collisions. Lee County, Florida, has led the state in watercraft-related mortality for eight of the last 10 years. This county is of particular concern as it contains important habitat for manatees, including extensive feeding grounds and an artificial warm-water refuge where more than 900 manatees have been recorded on a single day. Distributional aerial surveys were conducted from April 2007 through April 2009 over Lee County waters. Surveys yielded higher numbers of manatees than previously observed in this area. Using GIS methodology, kernel density analysis illustrated seasonal changes in distribution patterns and highlighted areas where manatees were most densely clustered. For example, during summer months, manatees were widely distributed throughout the survey area, with high-density areas associated with seagrass beds. During winter months, manatees were densely clustered at warm-water sites and over feeding grounds within close distance of these sites. These seasonal distribution patterns coincide well with speed zone designations. Counts and distributions of manatees were made available, almost immediately if necessary, to local marine law enforcement in an attempt to focus resources toward reducing manatee-watercraft collisions. Future studies should implement similar communication strategies to improve conservation efforts.     

Wait before running for your life: defensive tactics of spiny mice (<Emphasis Type=Italic>Acomys cahirinus</Emphasis>) in evading barn owl (<Emphasis Type=Italic>Tyto alba</Emphasis>) attack

Raptor–prey encounters were studied to evaluate the strategies and success rate of both predator attack and prey defense. We compared the success of barn owls in catching stationary simulated prey (food item) with that of moving prey (food item that was pulled in various directions). We also tracked real encounters between barn owls and spiny mice in a captive environment. It was found that owls had higher success in attacking stationary prey and that they seemed to attack the prey as soon as it became motionless. When attacked, only a few spiny mice remained immobile (freeze response) whereas most fled and usually avoided capture by the owls. It was also found that spiny mice displayed a preference to escape in those directions in which owls had demonstrated a lower success in catching the simulated prey. Escape initiation dichotomized to a short or long (but rarely intermediate) distance between the spiny mouse and the owl with more successful avoidance at short-distance (last-moment) escapes. The best predictor of escape success was the velocity of the spiny mouse, and the second best predictor was its flight initiation distance (FID). We present an update for Ydenberg and Dill’s model for optimal FID in close encounters, suggesting that fleeing at the last moment is advantageous. However, a last-moment attempt to escape is also more risky with a split second differing between life and death, and is therefore appropriate mainly for agile prey under close-distance attack.     

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.     

Inbreeding and caste-specific variation in immune defence in the ant <Emphasis Type=Italic>Formica exsecta</Emphasis>

Social insects are a widespread and ecologically dominant group. Reproductive division of labour among the females in the colonies is a key trait for their success, but at the same time, it creates dense aggregations of relatives which may promote the spread of disease in the colonies. Hence, the appropriate regulation of immune defence is crucial for the well-being of a colony. Inbreeding may disturb this process through reduced resistance or by impairing the colony’s ability to regulate the responses. We tested the effect of inbreeding and the within-colony differences in the encapsulation response between the two female castes of the ant Formica exsecta. New reproductive females (gynes) born in more inbred colonies, and being more inbred themselves, showed an elevated immune response whereas inbreeding had no effect on worker response. Furthermore, the immune response exhibited by gynes was lower than that of workers and was not dependent on their body size whereas the worker response increased with body size. The elevated response is likely to reflect genetic stress caused by inbreeding, which in turn may compromise colony founding and longevity. Indeed, eliciting a high immune response in itself might not be adaptive. Our results show that the regulation of the expression of immunity differs between female castes despite their similar genetic make-up.