Improvements to a passive trap for quantifying barnacle larval supply to semi-exposed rocky shores

Planktonic cyprid larvae of the intertidal barnacle, Semibalanus balanoides (L.) and the infralittoral/sublittoral Balanus crenatus Brug., are readily captured in small, chambered, passive traps deployed on rocky shores. Several structural developments of a previous trap design improved urea (killing solution) retention and capture of cyprids of both species. The design adopted for assessment of the larval supply/settlement relationship of the focal species, S. balanoides, included additional internal baffles and the replacement of the cylindrical trap opening with a coned aperture. Three sizes of cone aperture area were compared, with the intermediate size (1 cm²) being optimal for measuring larval supply over the full range of wave action on local shores. Most daily catches included larvae that were recognizable as having been dead before they were trapped, and had recently passed through the guts of grazing limpets (Patella vulgata L.). One site showed that ∼32% of trapped S. balanoides were so-called ‘faecal’ cyprids. An inability to distinguish these in trap samples would lead to a significant over-estimation of larval supply. Preliminary data indicated that S. balanoides larval supply to the intertidal is enhanced by onshore winds (or perhaps increased wave turbulence), whereas B. crenatus supply was positively correlated with offshore winds (or decreased turbulence). A clear ‘supply/settlement’ relationship for S. balanoides at six sites along ∼40 km of coastline was derived from 1 cm² coned trap catches and daily counts of settlement on ceramic tiles. There were no instances of disproportionately high settlement in relation to larval supply, but at five sites there were days when settlement apparently failed. The identification and explanation of settlement success and failure is fundamental to understanding the importance of larval supply in structuring marine benthic populations and communities.     

Effect of cyprid age on the settlement of balanus amphitrite darwin in response to natural biofilms

Larval settlement of the barnacle Balanus amphitrite Darwin (Cirripedia, Balanidae) is influenced by natural biofilms. In previous work by others, discriminatory settlement of aged cyprids has been observed in response to biofilms of different age. This study extends prior work by considering the effect of the age of cyprids on the outcome of settlement assays. Settlement was investigated with 0‐day‐old (newly metamorphosed) and 5‐day‐old cyprids. Biofilms under investigation were developed in the field for periods of 5 d and 1 month, and were subsequently included in laboratory settlement assays with a choice between a filmed and an unfilmed substratum. The bioassay was modified from the conventional horizontal dish design in order to generate a low water surface‐to‐volume ratio, which served to suppress larval entrapment in an organic layer on the water surface. Irrespective of cyprid age, a clear discrimination between a filmed and an unfilmed substrata was observed, and the preference for filmed or unfilmed substratum was dependent on the age of the cyprids. Settlement of 0‐day‐old cyprids was inhibited by a biofilmed substratum whereas induction occurred with aged cyprids. This pattern of settlement was independent of biofilm age. Bacterial abundance on unfilmed substrata in treatments and controls was significantly lower than that on biofilmed surfaces, confirming that bacterial contamination did not change the qualitative option during the assay.     

Consistent spatial patterns of arrival of larvae of the honeycomb barnacle Chamaesipho tasmanica Foster and Anderson in New South Wales

The small honeycomb barnacle Chamaesipho tasmanica occurs in patches at high levels on exposed rocky shores, but often carpets the substratum at mid-shore levels of sheltered shores in south-eastern Australia. Studies of larval supply from 1990 to 1993 and concurrent monitoring of settlement from 1991 to 1993 revealed that larval arrival and settlement were typified by trickles of larvae from late July to December (although some were observed in January and February). Major pulses of arriving cyprids were also recorded once or twice each year. While local patterns of water-flow had no impact on numbers of larvae arriving, major peaks of larval arrival were always associated with strong southerly winds during new and full moons. There was a consistent spatial pattern of larval supply; more larvae were always caught in one area low on the shore. Numbers of larvae caught were, however, very sporadic within a given year and very variable from one year to the next. While the different numbers of cyprids in different places cannot be explained by cyprids arriving first on lower parts of the shore, longer periods of submersion nor aggregations of larvae in the plankton, recurrent patterns of arrival of larvae suggest that local site-specific characteristics have an influence on the demography of populations of this species. Variations in numbers of larvae arriving were responsible for the variations in distributions of juveniles on the substratum.     

The role of settlement in structuring a hard-substratum community in the deep sea

Larval settlement can play an important role in the structure of benthic communities in shallow-water habitats. Experimental studies of settlement and colonization in the deep sea, however, are few, and have concentrated exclusively on sedimentary habitats. Results are presented here from an in situ study designed to determine whether larval settlement patterns on a common, deep-sea hard substratum (manganese nodules) determine the subsequent structure of the epifaunal community. In a 7-wk deployment at a depth of 1240 m, settlement onto manganese nodules resting on the sediment in a relatively “low-flow” environment was significantly greater than onto nodules suspended in a “higher-flow” regime. After 2 yr, this difference in total number of individuals was no longer significant, but a new pattern in community composition emerged, which correlated to the presumed supply of paniculate food. Suspension-feeders dominated the elevated nodules (correlating to higher horizontal participate fluxes) and deposit-feeders dominated the nodules sitting on the sediment (correlating to higher particulate deposition). The faunal settlement patterns observed after 7 wk, therefore, appear to be altered by adult feeding requirements of the component species.     

Barnacle larval supply to sheltered rocky shores: a limiting factor?

In northwest Europe, sheltered rocky shores are dominated by fucoid canopy algae and barnacles are rare, although the latter are extremely abundant on exposed shores. The supply of the intertidal barnacle Semibalanus balanoides (L.) to sheltered, fucoid dominated rocky shores was investigated to determine the importance of larval supply in limiting the abundance of adults in shelter. Larval supply was measured at two spatial scales, at the scale of shore (100s of metres), by comparing larval concentrations at exposed and sheltered sites, and at a smaller spatial scale (m), by examining the role of fucoid canopies in limiting supply to the substratum. Replicate plankton trawls were carried out above the intertidal zone at high water at two sheltered sites and nearby exposed headlands. The concentration of S. balanoides cyprid larvae was significantly higher at the sheltered sites on two out of three sampling occasions with up to 14 times greater larvae on one occasion than the nearby exposed site. The effect of the macroalgal canopy on supply to the substratum was assessed in two ways: directly, by pumping water from the substratum in areas with and without Ascophyllum nodosum (L.) Le Jolis, and indirectly by measuring cyprid settlement in a canopy-manipulation experiment. Pumped plankton samples from mid tide level showed that the A. nodosum canopy did not form a barrier to larval supply and may have had a positive effect on larval concentrations at the substratum. Cyprid settlement was assessed in the mid shore A. nodosum and low shore Fucus serratus L. zones to areas with canopy algae (but protected from the sweeping effects of macroalgal fronds) and without canopy. Settlement over three consecutive 24-h periods showed a consistent pattern; settlement was consistently lower beneath the F. serratus canopy than in cleared areas, suggesting that this algal species forms a barrier, limiting supply of cyprid larvae to the substratum.     

Cochliomycin A inhibits the larval settlement of Amphibalanus amphitrite by activating the NO/cGMP pathway

Cochliomycin A is a compound with anti-barnacle settlement activity and low toxicity, but the molecular mechanism of the compound is unknown. Here, isobaric tags for the relative or absolute quantitation (iTRAQ) labeling proteomic method were applied to analyze changes in the proteome of Amphibalanus（=Balanus） amphitrite cyprids in response to cochliomycin A treatment. Cochliomycin A affected the cytochrome P450, glutathione S-transferase (GST) and NO/cGMP pathways, among which the NO/cGMP pathway was considered to play a key role in barnacle larval settlement, while the cytochrome P450 and the GST pathways are mainly for detoxification. The results of real-time PCR further suggested the NO/cGMP pathway was activated in response to cochliomycin A. Larval settlement assays revealed that S-methylisothiourea sulfate (SMIS) and 1H-(1,2,4)oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) rescued cyprids from cochliomycin A-induced inhibition of larval settlement. The findings supported the hypothesis that cochliomycin A inhibited barnacle larval settlement by stimulating the NO/cGMP pathway.     

The interplay of substrate nature and biofilm formation in regulating Balanus amphitrite Darwin, 1854 larval settlement

The settlement of marine larvae is influenced by a wide range of physical and biological factors. It is still poorly known how the nature of substrate and the biofilm can interact in regulating settlement patterns of invertebrate larvae. Here we use laboratory experiments focused on settlement behaviour of the barnacle Balanus amphitrite. The aim of this work is to understand whether: (i) the nature of substratum can affect biofilm formation and its structure, (ii) the nature of substratum can affect B. amphitrite larval settlement, (iii) the age of the biofilms and the nature of substrate can interact in influencing larval settlement.Four kinds of substrata (marble, quartz, glass, and cembonit) were biofilmed under laboratory conditions for 5, 10 and 20 days at the temperature of 28 °C. Settlement response was investigated with 5-day-old cyprids. Biofilms were quantitatively and qualitatively analysed by scanning electron microscopy. The settlement of B. amphitrite larvae significantly differed among substrata; also, the patterns of development of biofilm assemblages changed with substrate. In addition, the larval attractiveness of different substrates tends to disappear with biofilm age.     

Larval development and settlement of a whale barnacle

Larval development and settlement of whale barnacles have not previously been described, unlike intertidal barnacles. Indeed, the mechanisms of the association between barnacles and whales have not been studied. Here we describe the larval development and settlement of the whale barnacle, Coronula diadema, and possible involvement of a cue from the host in inducing larval settlement. Eight-cell stage embryos were collected from C. diadema on a stranded humpback whale, incubated in filtered seawater for 7 days, and nauplius larvae hatched out. When fed with Chaetoceros gracilis, the nauplii developed to stage VI, and finally metamorphosed to the cypris stage. The larval development looked similar to that of intertidal barnacles with planktotrophic larval stages. The cyprids did not settle in normal seawater, but did settle in polystyrene Petri dishes when incubated in seawater with a small piece of skin tissue from the host whale. This strongly suggests the involvement of a chemical cue from the host whale tissue to induce larval settlement.     

藤壶金星幼虫附着变态机制

藤壶属节肢动物门(Arthropoda)甲壳亚门(Crustacea)蔓足下纲(Cirripedia)围胸总目(Thoracica), 具备特殊的形态结构、生活史和种群生态特征,是最主要的海洋污损生物。其幼虫阶段通常经历6期无节幼体和1期不摄食的金星幼虫,从浮游的金星幼虫附着变态成固着的稚体是藤壶生活史中的一个关键环节。外界化学和生物因子中成体提取物、水溶性信息素、足迹、神经递质、激素、生物膜等均影响藤壶金星幼虫的附着变态;内在因子即金星幼虫的生理状态(能量储量和年龄)决定了其对外界因子的反应程度。概括了近年来藤壶附着变态生理机制和分子机制研究的进展,可为深入了解藤壶金星幼虫附着变态机制提供参考,也为开发新型、高效、环保的防污剂提供理论指导。     

Larval densities and a life-table for the gypsy moth, Lymantria dispar, estimated using the head-capsule collection method

ABSTRACT. 1. In order to estimate the absolute larval density in each stage of a larval population of the gypsy moth, Lymantria dispar L., in a deciduous forest in northern Japan, the head-capsule collection method was used. An estimate by this method was compared with estimates based on two kinds of frass collection methods.
2. Twenty-one traps made of cloth were put in a study plot. Each trap was of 24.5 cm diameter. Larval head-capsules falling into the traps were collected and sorted by hand. On the first sampling occasion, the population was also estimated using the frass-collection method.
3. Larval numbers estimated by the head-capsule collection method were almost identical to estimates by the two frass methods. Larval numbers entering the four larval instar were successfully estimated by the head-capsule collection method, and an age-specific life table was established using the resultant estimates.     

Larval supply is a good predictor of recruitment in endemic but not non-endemic fish populations at a high latitude coral reef

Despite extensive research, factors influencing the importance of pre- and post-settlement processes to recruitment variability remain ambiguous. Using a novel perspective, we investigated the potential influence of endemism on the relationship between larval supply and recruitment in reef fish populations at Lord Howe Island, Australia. Larval supply and recruitment were measured for three regional endemic and four widespread non-endemic species using light traps, artificial collectors, and underwater visual censuses. Recruitment was correlated with larval supply in endemics but not in non-endemics, likely due to a combination of low larval supply and post-settlement survival of non-endemics. Surveys also indicated that endemics were far more abundant and occurred in more locations than closely related non-endemics. These preliminary findings suggest that either local adaptation enhances recruitment in endemics through higher larval replenishment rates or reduced post-settlement mortality, populations of widespread species at the periphery of their range are poorly adapted to local environmental conditions and therefore experience lower and more variable settlement and post-settlement survival rates, or both.     

Larval recruitment of the blue mussel Mytilus edulis: The effect of flow and algae

The mussel Mytilus edulis settlement and distribution was studied on plastic panels with manipulated flow regime (faired, bluff, split and angled) with or without water soluble metabolites of the green alga Cladophora rupestris. The panels were exposed vertically on a device (hydrovane) that ensures their constant orientation in the current during the peak of larval settlement at 1 m depth. In order to investigate larval distribution on the panels, half of them were coated with a silicone vacuum grease that prevents larvae from de-attachment. This grease was not toxic and did not attract or repel larvae. Low densities of larvae on the un-greased plates compared to the greased ones suggested that some of larvae left the substratum. The blue mussel larvae initially settled in regions of reduced shear velocity and then redistribute to the regions of high shear velocity. The presence of the alga increased the density of blue mussel larvae and changed their distribution on the panels. Overall, our results demonstrated that larval recruitment of M. edulis is an active process affected both by boundary-layer hydrodynamics and algal waterborne compounds.     

Probing the settlement signals of Amphibalanus amphitrite

To achieve their reproductive potential, barnacles combine tactile exploration of surface structural properties and integration of cellular signals originating from their antennular sensory setae within a developmentally defined, temporally narrow window of settlement opportunity. Behavioural assays with cyprids coupled with biometric analysis of scanning electron microscopy-acquired images in the presence of specific chemical compounds were used to investigate how settlement on a substratum is altered in response to the presence of these compounds. Impeding tactile exploration was shown which altered cellular signalling and/or induced malformation of anatomical features of the antennular sensory setae, which disrupted the settlement behaviour of the model barnacle species Amphibalanus amphitrite. It is concluded that surface exploration by the cyprids relies on mechanical and nociception-related and calcium-mediated signals while a protein kinase C signalling cascade controls the timely metamorphosis of the cyprids to sessile juveniles.     

Expression of calmodulin and myosin light chain kinase during larval settlement of the Barnacle Balanus amphitrite

Barnacles are one of the most common organisms in intertidal areas. Their life cycle includes seven free-swimming larval stages and sessile juvenile and adult stages. The transition from the swimming to the sessile stages, referred to as larval settlement, is crucial for their survivor success and subsequent population distribution. In this study, we focused on the involvement of calmodulin (CaM) and its binding proteins in the larval settlement of the barnacle, Balanus ( = Amphibalanus) amphitrite. The full length of CaM gene was cloned from stage II nauplii of B. amphitrite (referred to as Ba-CaM), encoding 149 amino acid residues that share a high similarity with published CaMs in other organisms. Quantitative real-time PCR showed that Ba-CaM was highly expressed in cyprids, the stage at which swimming larvae are competent to attach and undergo metamorphosis. In situ hybridization revealed that the expressed Ba-CaM gene was localized in compound eyes, posterior ganglion and cement glands, all of which may have essential functions during larval settlement. Larval settlement assays showed that both the CaM inhibitor compound 48/80 and the CaM-dependent myosin light chain kinase (MLCK) inhibitor ML-7 effectively blocked barnacle larval settlement, whereas Ca(2+)/CaM-dependent kinase II (CaMKII) inhibitors did not show any clear effects. The subsequent real-time PCR assay showed a higher expression level of Ba-MLCK gene in larval stages than in adults, suggesting an important role of Ba-MLCK gene in larval development and competency. Overall, the results suggest that CaM and CaM-dependent MLCK function during larval settlement of B. amphitrite.     

Dynamics and extent of larval lake sturgeon Acipenser fulvescens drift in the Upper Black River, Michigan

Lake sturgeon larval drift is not uniform in time or space and subsequent efforts to determine the relative abundance have suffered because of the lack of information during this early life history period. The purpose of this study was to obtain information about the early life history of lake sturgeon, determine the extent and duration of lake sturgeon larval drift, and examine this relationship to water flow and temperature in the Upper Black River, Michigan. This study also compares the results of other studies to further evaluate the dispersion of larvae. Larval production was quantified using drift nets anchored to the stream bottom from May to June in 2000–2002. Larval drift nets captured 780 larvae in 2000; 2975 larvae in 2001; and 2041 larvae in 2002. For the 2000, 2001, and 2002 spawning season, we estimated that 7107 (95% CL: ± 1470), 17 409 (95% CL: ± 5163), and 15 820 (95% CL: ± 3168) larval lake sturgeon were produced in the Upper Black River (UBR), respectively. Catch per unit effort values of drifting larvae were greatest after peak water flows, with most larvae captured in the middle of the river channel. A mean daily water temperature above 16°C was an important environmental stimulus that influenced peak larval dispersion away from spawning sites. The results of this study suggested that natural reproduction was still occurring in the Black Lake system.     

Influence of trap type, trap colour, and trapping location on the capture of the pine moth, Thaumetopoea pityocampa

Studies were conducted to evaluate the effect of type, location, and colour of traps baited with 1 mg of the sex pheromone (Z)‐13‐hexadecen‐11‐ynyl acetate in polyethylene vials on the capture of the pine processionary moth, Thaumetopoea pityocampa (Denis & Schiffermüller) (Lepidoptera: Thaumetopoeidae), males. The experiments were carried out during two flight seasons in 2002 and 2003 at a low elevation Mediterranean pine forest on the hill of Goritsa (Magnesia, Thessaly, Central Greece). The hill is covered by approximately 120 ha of pines, among which Pinus brutia Ten. (Pinaceae) is predominant with Pinus halepensis Mill. (Pinaceae) as second most common species. Among the commercially available trap types used, the Delta and Pherocon II captured significantly more males than the Funnel trap. Furthermore, pine density had a significant effect on trap type and trap colour catches. Delta traps caught significantly more adults than the other two trap types in the low density pine stand, while in the medium and high‐density pine stands no significant differences were noted between Delta and Pherocon II traps. In addition, trap colour performance varied according to pine density; white‐ and yellow‐coloured traps caught significantly more males than the other two trap colours in the high‐density stand. The first adults were captured in traps during May, and traps continued to capture adults in low numbers (< two individuals per trap) until mid‐June. A period of 3–4 weeks of no trap captures followed until mid‐July, when captures restarted, at low numbers. The peak of T. pityocampa flight was observed during late August–September.     

The use of computer-assisted motion analysis for quantitative studies of the behaviour of barnacle (Balanus amphitrite) larvae

The effects of larval density and age on pre-settlement swimming behaviour of Balanus amphitrite cyprid larvae were studied with the aid of computer-assisted motion analysis. Swimming behaviour was monitored in individual, in groups of 10-15 and in groups of 50-100 cyprids. There was a small, but significant effect of larval density on swimming speed and no effect on two other quantitative measures: rate of change of direction and net-to-gross displacement ratio. There was also small but significant variation in swimming speed between different batches of cyprids over the course of 2 years. Swimming behaviour of individual cyprid larvae was also monitored daily for 7 days, with the larvae maintained in the cold and dark between measurements to prevent settlement and metamorphosis. There were no significant behavioural differences observed over time indicating that larvae may be held in this manner experimentally without affecting these parameters.     

An ordinal day model of spruce beetle trap capture phenology in northern Colorado

Spruce beetle populations (Dendroctonus rufipennis) (Coleoptera: Scolytinae) in Engelmann spruce (Picea engelmannii) stands were monitored at twelve sites for 6 years in northern Colorado using pheromone‐baited Lindgren funnel traps. During the trapping period, over 30,000 beetles were captured, and beetle abundance data were used to construct empirical models of trap capture phenology based on day of year, accumulation of thermal units and thermal day thresholds, with the goal of informing future regional trapping efforts and producing a simple model for use by forest managers. Ordinal day models outperformed thermal accumulation and thermal threshold models in both predictive power and parsimony. Mean date of earliest capture was Day 153 (June 2) and ranged from Day 126 to Day 161 (May 6–June 10), and mean date of final capture was Day 243 (August 31) and ranged from Day 220 to Day 286 (August 8–October 13). A two‐parameter logistic function was the most parsimonious of several ordinal day models, accounting for 81% of the variance in cumulative trap capture across all sites and dates. The model predicts 50% of trap captures to occur by Day 181 (June 30). This model has application as a decision support tool for forest ecosystem managers concerned with the timing of trap deployment or D. rufipennis mitigation treatments.     

Gregarious settlement in cypris larvae:the effects of cyprid age and assay duration

Gregarious behaviour of marine larvae is perhaps most clearly associated with finding a suitable habitat in a changeable or restricted environment, or with finding other conspecifics with which to mate. Prior work has shown that in settlement assays using cypris larvae of the barnacle Balanus amphitrite, gregarious interactions significantly affected the interpretation of experiments testing the activity of organic settlement promoters and inhibitors. Other studies have also shown effects of cyprid age and pheromone concentration on settlement behaviour. However, the effects of interactions between gregariousness and these two factors are not known. The aim of this study was to test the hypotheses that i) as cyprids age the effects of gregariousness become less apparent, and ii) as the duration of the experiment increases gregarious effects become more apparent, using cypris larvae of B. amphitrite and Balanus improvisus. Three age classes of cyprids were used at six densities in a fully factorial design. For B. improvisus cyprids significant gregarious effects occurred between 3 or more larvae, and although larval age and experiment duration had significant main effects, there were no interactions between these important factors and gregariousness. For B. amphitrite cyprids significant gregarious effects also occurred with 3 larvae per well, though this effect was strongly dependent upon experiment duration. B. amphitrite cyprid sensitivity to conspecific cues does not change with age, although increasing experiment duration and age interact to increase settlement. Differences between species may be due to different thresholds to conspecific larval cues, or B. improvisus cyprids release much more larval temporary adhesive during exploration.