Role of protein in larval swimming and metamorphosis of Bugula neritina (Bryozoa: Cheilostomatida)

Larvae of the marine cheilostomatid bryozoan Bugula neritina (L.) were prevented from settling for 1, 4 and 8 h by mechanical agitation, following which settlement and metamorphosis success were examined. Settlement rates were significantly affected by swimming time, which decreased from 100% after 2 h to 93.7 ± 4.3% after 8 h. Similarly, metamorphosis to the feeding ancestrula was significantly impaired following a swimming time of 8 h, declining from 93.7 ± 4.3% after 1 h to 65.9 ± 7.0% after 8 h. The resultant colonies grew well for the first 3 wk, following which time, growth patterns became erratic. Growth rate was in all cases highly variable, and did not correlate with enforced swimming times. Larval protein composition was examined after 1, 4 and 8 h swimming time, and post-larval composition 1, 2, 5, 24 and 48 h after settlement using sodium-dodecyl-sulphate polyacrylamide-gel electrophoresis (SDS-PAGE). Individual protein content was measured using a densitometer. Larvae did not consume protein during swimming, however a protein measuring 170 kdaltons was consumed during metamorphosis. These results are discussed in the context of larval settlement and energetics. Received: 19 July 1998 / Accepted: 3 December 1998     

Rates of energy consumption and acquisition by lecithotrophic larvae of Bugula neritina (Bryozoa: Cheilostomata)

Lecithotrophic larvae of the cheilostome bryozoan, Bugula neritina (L.), lose metamorphic competence 12 to 24 h after release from the maternal zooid. The high respiration rate of newly released larvae (mean=306.3 pmol O₂ larva^-1 h^-1, range=149.3 to 466.6, n=18 trials, 22.5°C) from adults collected at Link Port, Fort Pierce, Florida during the winter/spring of 1990–1991 reflects their active swimming behavior. The average energy content per larva was 15.24 mJ (range: 13.35 to 20.17 mJ ind^-1, n=5 groups). If all cells have an identical energy content and metabolic rate, then 2 and 20% of the total energy content would be consumed by the onset (2 h post-release) and the loss (24 h post-release) of metamorphic competence. Larvae of B. neritina are a composite of both larval and juvenile tissues and the loss of metamorphic competence may be due to regional depletion of labile energy stores in transitory larval cells, particularly the ciliated cells that comprise the locomotory organ, the corona. Although nonfeeding, B. neritina larvae can acquire nutrients from the environment in the form of dissolved organic materials (DOM) in seawater. Both the amino acid alanine and the fatty acid palmitic acid can be transported from seawater ([S]=1 M, 22.5°C). The rates of alanine influx (appearance of label in tissue) averaged 0.366 pmol larva^-1 h^-1 and, based on comparisons between rates of solute transport and metabolism, would contribute little (<1% of required energy) to offset the metabolic demand. The average rate of palmitic acid influx was 4.668 pmol larva^-1 h^-1 and assuming that the measured influx equals the net solute flux, could account for 21 to 72% of energy requirements. These data suggest that the duration of planktonic life of B. neritina larvae is principally regulated by the amount of endogenous energy stores, but may be modulated by available DOM in seawater.     

Localisation of biogenic amines in larvae of Bugula neritina (Bryozoa: Cheilostomatida) and their effects on settlement

Distributions of serotonin and catecholamines in larvae of the marine bryozoan Bugula neritina (Bryozoa: Cheilostomatida) were investigated using immunohistochemistry with anti-serotonin antiserum and glyoxylic acid–induced fluorescence histochemistry. Anti-serotonin immunoreactive substances and glyoxylic acid–induced fluorescent substances had similar distributions in the equatorial neuromuscular ring, the neural plexus, the paired axial neuromuscular cords, and tracts connecting the neural plexus to ciliated cells bordering the pyriform organ. The effects of dopamine, noradrenaline, adrenaline, tyramine, octopamine, synephrine and serotonin, at 10⁻⁴, 10⁻⁵ and 10⁻⁶  M, on settlement were analysed. In filtered seawater, 98% of larvae settled in 3 h, but only 11%, 3% and 6% total settlement was observed after 8 h in 10⁻⁴  M dopamine, 10⁻⁴  M serotonin and 10⁻⁵  M serotonin, respectively. Total settlement was 70% in 10⁻⁴  M noradrenaline, 80% in 10⁻⁴  M adrenaline and 60% in 10⁻⁴  M tyramine. Less than 60% settlement was observed in 10⁻⁴ and 10⁻⁵  M octopamine and synephrine. Serotonin's inhibitory effect on settlement was mimicked by a range of serotonin receptor agonists and antagonists, among which 5-carboxamidotryptamine was the most potent. Received: 19 March 1999 / Accepted: 11 October 1999     

The influence of seawater temperature on zooid size and growth rate in Pentapora fascialis (Bryozoa: Cheilostomata)

Zooid size variation and growth banding in the bryozoan Pentapora fascialis, collected from the South of Great Britain to the South of Italy, were investigated in order to test their relationship with the variation of seawater temperature. Zooid length appeared to be more sensitive than zooid area and width to temperature variation. The longest zooids were budded in localities that typically experience low seasonal variation in temperature (3.2–7.5°C) and low mean annual temperature (about 11°C). Zooid length demonstrated the strongest relationship with the range of seasonal variation in temperature rather than the mean annual temperature. Growth bands produced during colder months were less high than those produced during warmer months. Measurements of these alternating bands were used to calculate growth rate of the laminae, ranging from 20.7 at Plymouth to 29.3 mm year⁻¹ at Tino Island. Significant differences in zooid length were found between the two bands, with longer zooids within the band developed in colder months and shorter zooids in bands produced in the warmer period. We conclude that zooid size can be used as an environmental proxy of the seasonal temperature regimes experienced by the species.     

Availability of dissolved organic matter offsets metabolic costs of a protracted larval period for <Emphasis Type="Italic">Bugula neritina</Emphasis> (Bryozoa)

For nearly a century researchers have investigated the uptake and utilization of dissolved organic matter (DOM) by marine invertebrates, but its contribution to their growth, reproduction, and survival remains unclear. Here, the benefit of DOM uptake was assessed for the marine bryozoan Bugula neritina (Linnaeus 1758) through performance comparisons of individuals in the presence and absence of DOM. The experiments were performed using B. neritina collected from floating docks in Beaufort, NC, USA from July to September 2004. Seawater was subjected to ultraviolet irradiation to reduce naturally occurring DOM, and then enriched with either 1 μM of palmitic acid or a mixture containing 1 μM each of glucose, alanine, aspartic acid and glycine. Larvae in DOM-enriched and DOM-reduced treatments were sampled and induced to metamorphose following 1, 6, 12, and 24 h of continuous swimming at 25°C. Sampled larvae were assessed for initiation of metamorphosis, completion of metamorphosis, and ancestrular lophophore size to determine the extent to which energy acquired from DOM uptake could offset the metabolic costs of prolonged larval swimming. DOM treatment had no significant effect on initiation of metamorphosis, but did have a significant effect on completion of metamorphosis and lophophore size. Larvae swimming in DOM-enriched treatments for 24 h experienced a 20% increase in metamorphic completion rate, compared to larvae swimming for 24 h in the DOM-reduced treatment. In addition, larvae in the amino acid and sugar mixture for 24 h had a significantly larger lophophore surface area and volume (23 and 31%, respectively), compared to larvae in DOM-depleted seawater. To ensure that the increases in performance found in larvae with access to DOM were not due to a decrease in metabolic activity, the respiration rates for these larvae were compared to those of larvae in DOM-depleted seawater. There were no significant differences between these treatments, indicating that the increases in performance were due to the energy acquired from DOM. These results clearly show that for B. neritina, DOM uptake results in increased metamorphic success and in the size of the feeding apparatus following an extended larval swimming duration.     

Effect of microbial films on settlement of bryozoan larvae (Bugula simplex,B. stolonifera and B. turrita)

The influence of microbial films on larval settlement was studied for sympatric species of cellularioid cheilostome bryozoans (Bugula simplex, B. stolonifera, and B. turrita). Settlement experiments were conducted in the laboratory in 1981 with and without a choice of filmed or unfilmed substrates. When given a choice of substrates, the larvae of all three bryozoan species significantly preferred the filmed substrate. When not given a choice of substrates, both B. simplex and B. turrita larvae tended not to settle on the unfilmed substrate, while the filmed and unfilmed substrates were equally attractive to B. stolonifera. Microbial films enhanced the settlement of all three species.     

Effects of temperature and salinity on larval development ofNecora puber (Brachyura: Portunidae)

Temperature and salinity affected both length of larval development and mortality inNecora puber collected in the Ría de A Coruña during December 1984 and January 1985. Development time decreased considerably with increased temperature. This decrease was sharper when temperature increased from 15° to 20°C than when it increased from 20° to 25°C. At 35S, average development took 48, 32 and 28 d at 15°, 20° and 25°C, respectively. At the three salinities tested (25, 30 and 35), larval development was completed only at 15°C, at 20°C/30 and 35S, and at 25°C/35S. Development times at 15° and 20°C were highly significantly different at both 35 and 30S (P 0.01). However, there were no significant differences between development times at 20° and 25°C (P > 0.05). Within any one specific temperature series, no significant difference was observed between the salinity values tested (P > 0.05). The duration of each of the five zoeal stages was similar within each and the same temperature/salinity combination, whereas the duration of the megalop was twice as long as any of the zoeal stages. The combination of the lowest temperature (15°C) and the highest salinity (35) tested resulted in the greatest larval survival of 28%. Highest mortality occurred at 25°C, at which temperature development was completed only at 35S. A sharp drop in larval survival was observed in the transition period Zoea V — megalop in all combinations of temperature and salinity tested. Within the limits of tolerance to temperature and salinity, the former effected more pronounced differences in the duration of larval development, while salinity appeared to constitute a limiting factor for survival.     

Effects of high temperature on larval development and metamorphosis of Arachnoides placenta (Echinodermata: Echinoidea)

Adults of the sea urchin Arachnoides placenta (L.) were induced to spawn, and eggs were fertilized at 28°C in September 1989. After 5 min, eggs were transferred to 28, 31, 34, or 37°C and reared to metamorphosis. Embryos were observed at 20-min intervals during the first 2 h; larvae were observed daily. The cleavage was higher at higher temperatures. Embryos reared at 28°C were still at the 4th cleavage (16-cell stage) after 100 min, while those at 34°C had reached the 5th cleavage (32-cell stage). All embryos reared at 37°C died on the second day. Incidence of abnormality was 20 to 30% at 28 and 31°C, 48% at 34°C, and 77% at 37°C. The 8-arm stage was reached after 4 d at 28°C, 3 d at 31°C and 2 d at 34°C. Larvae displayed decreasing body length and arm length with increasing temperature. Larvae at 31°C have relatively long arms, as a result of a decrease in body length, not because of increased arm length. Incidence of metamorphosis was 43.9±1.7% (mean/plusmn;SD) at 28°C, 24.5±1.9% at 31°C, and 5.3% at 34°C. The size of metamorphosed juveniles was significantly larger at 28°C than at 31 and 34°C. Temperatures of 31°C negatively affect larvae and juveniles of the sand dollar.     

Effects of temperature and salinity acclimation of adults on larval survival,physiology, and early development of Lytechinus variegatus (Echinodermata: Echinoidea)

Larval survival and developmental rates of Lytechinus variegatus (Lamarck) were determined as a function of temperature and salinity in two experiments by: (1) directly transferring fertilized eggs to 35, 30, 27.5, 25, 20, 15, and 10S seawater at 18 and 23°C, and (2) acclimation of adult sea urchins to the conditions described above for 1 to 4 wk prior to spawning. Developmental rates and percent survival of larvae prior to metamorphosis decreased at salinities below 35 (Q₁₀ values for metamorphosis=0.380 to 0.384). Temperature and salinity significantly (P<0.05) affected metabolic rates of L. variegatus plutei. These results show that L. variegatus larvae are stenohaline when compared to larvae of other echinoderm species. LC₅₀ values (S), developmental rates, and survival to metamorphosis indicate that acclimation of adult sea urchins to lower salinity prior to spawing and fertilization does not enhance development or survival of embryos exposed to low salinity.     

Reproduction and larval development of Marenzelleria viridis (Polychaeta: Spionidae)

The pelagic larvae of the polychaete Marenzelleria viridis (Verrill, 1873) are newcomers among the meroplankton of the Baltic Sea, where they sometimes achieve an abundance of up to 21x10⁶ ind m^-3 near the coast, especially in late autumn and even in winter. Benthos samples were collected in the Darss-Zingst bodden chain from April 1992 to 1993 and used to ascertain the reproductive stages of individuals. Development from fertilized egg to benthic juvenile is described on the basis of field material. Details of the morphology of various pelagic stages and the young benthic worm are presented as drawings and scanning electron microscope photographs. In 1992, development of the gametes started in mid-May. The individuals reached maturity in late September after about 20 wk. The first planktonic stage was the fertilized egg. The larvae have initially one and later two pairs of black eyes and, in the 2nd segment, a ciliated pit. Gastrotrochs are present on the third and thereafter every alternate segment. Neuropodial setae develop once the 7-setiger stage is reached. Palps appear at the 10-setiger stage, and neuropodial bidentate hooks from the 10th to 11th setiger on. Metamorphosis into the juvenile benthic stage takes place at the earliest when the 15-setiger stage has been reached. The reproductive seasons of various populations of M. viridis and the developmental patterns of their larvae are discussed, and differences are compared with relevant findings for the spionids.     

Rate of growth and longevity of a large colony of Pentapora foliacea (Bryozoa) recorded in their oxygen isotope profiles

We studied the growth patterns and the stable oxygen isotopic composition of an individual of Pentapora foliacea (Ellis and Solander 1786) collected on October 6, 1978 from the Bristol Channel, Pembrokeshire, UK, Irish Sea. The stable oxygen isotopes are in equilibrium with the ambient seawater and show a marked seasonal variation reflecting seasonal water temperature changes. The stable oxygen isotopes further suggest that regular growth patterns of less calcified growth bands, which are secreted in winter, are perennial. These winter growth check lines can easily be used to determine longevity of these bryozoa colonies and of annual growth rates. The colony analysed is at least 3-years-old with an annual growth rate of approximately 2 cm per year.     

Effects of clotrimazole and amiodarone on early development of amphibian (Xenopus tropicalis)

Pharmaceuticals have been recognized as a continuing threat to environmental stability. Few experimental data are available for the effects of clotrimazole and amiodarone on the ecological environment. An acute test with embryos and a chronic test with larvae of amphibian (Xenopus tropicalis) were thus conducted to determine the influence of clotrimazole or amiodarone on early amphibian development. In acute test, % survival and the body length were numerically decreased by both pharmaceuticals treatments compared to control. In chronic test, the cumulative mortality was 22.2% with 0.1?µg?L^?1 clotrimazole treatment and 21.7% with 1?µg?L^?1 amiodarone. The whole body length and the biomass were significantly decreased and developmental stages significantly delayed by both pharmaceuticals. The results of our study suggest that clotrimazole exerted adverse effects on larvae of X. tropicalis at environmentally relevant concentrations.     

Reproductive behaviour and larval development of Tanais cavolinii (Crustacea: Tanaidacea)

Mature female stages of Tanais cavolinii Milne Edwards are described according to degree of oostegite development. Sexual dimorphism of the chelipeds of copulatory males is explained functionally. An aberrant marsupium consisting of two closed ovisacs with a small slit for sperm transfer protects the brood from sudden unpredictable changes in salinity. Behavioural traits, courtship, copulation, and parental care are described and compared to those of Heterotanais oerstedi Kröyer. The different habitats selected by the two species are reflected by several adaptations in their morphology and behaviour as tube dwellers. A phenomenon unique among invertebrates is that the female T. cavolinii supplies additional yolk to the larvae just before their release. The production and timing of this food supply is described and its adaptive significance is discussed.     

Effects of colony food shortage on behavioral development in honey bees

Three experiments were conducted to explore the effects of severe food shortage on the control of two important and interrelated aspects of temporal division of labor in colonies of the honey bee (Apis mellifera): the size and age distribution of a colony's foraging force. The experiments were conducted with single-cohort colonies, composed entirely of young bees, allowing us to quickly distinguish the development of new (precocious) foragers from increases in activity of bees already competent to forage. In experiment 1, colony food shortage caused an acceleration of behavioral development; a significantly greater proportion of bees from starved colonies than from fed colonies became precocious foragers, and at significantly younger ages. Temporal aspects of this starvation effect were further explored in experiment 2 by feeding colonies that we initially starved, and starving colonies that we initially fed. There was a significant decrease in the number of new foragers in starved colonies that were fed, detected 1 day after feeding. There also was a significant increase in the number of new foragers in fed colonies that were starved, but only after a 2-day lag. These results suggest that colony nutritional status does affect long-term behavioral development, rather than only modulate the activity of bees already competent to forage. In experiment 3, we uncoupled the nutritional status of a colony from that of the individual colony members. The behavior of fed individuals in starved colonies was indistinguishable from that of bees in fed colonies, but significantly different from that of bees in starved colonies, in terms of both the number and age distribution of foragers. These results demonstrate that effects of starvation on temporal polyethism are not mediated by the most obvious possible worker-nest interaction: a direct interaction with colony food stores. This is consistent with previous findings suggesting the importance of worker-worker interactions in the regulation of temporal polyethism in honey bees as well as other social insects. Received: 17 April 1997 / Accepted after revision: 26 December 1997     

Prolonged exposure to low dissolved oxygen affects early development and swimming behaviour in the gastropod Nassarius festivus (Nassariidae)

Effects of low dissolved oxygen on early development and swimming behaviour of veliger larvae of the scavenging gastropod Nassarius festivus were studied. Embryonic development was significantly delayed when dissolved oxygen level was reduced to 3.0 mg O₂ l⁻¹ and no embryo hatched successfully at 0.5 mg O₂ l⁻¹. Veliger larvae hatched at 4.5 mg O₂ l⁻¹ had significantly smaller velar lobe, shell length and shell width. Median 48-h LC₅₀ value of the veliger larvae was estimated at 1.25 mg O₂ l⁻¹ with lower swimming speed (swimming velocity and dispersal velocity) being recorded for the survivors exposed to reduced oxygen levels. The percentage of veliger larvae that developed into crawling juveniles was significantly reduced and metamorphosis was delayed at 4.5 mg O₂ l⁻¹ whereas all larvae at 3.5 mg O₂ l⁻¹ died before they underwent metamorphosis. Juveniles developed at 4.5 mg O₂ l⁻¹ were also smaller than those at 6.0 mg O₂ l⁻¹. Results indicated that dissolved oxygen levels well above hypoxia levels (2.8 mg O₂ l⁻¹) have already had significant impact on the hatching success and larval development in gastropods, which may lead to long-term decreases in population growth.     

Effects of the insect growth regulator Dimilin® (TH 6040) on larval development of two estuarine crabs

Effects of Dimilin^® (TH 6040), an insect growth regulator which interferes with the formation of the insect cuticle, were studied on the larval development of Rhithropanopeus harrisii (Gould) and Sesarma reticulatum (Say) (Crustacea: Brachyura). When larvae were exposed to 0.5 (R. harrisii only), 1, 3, 5, 7, and 10 ppb Dimilin from hatching to the first crab stage, survival in both species decreased in relation to increased concentrations of Dimilin. Survival of R. harrisii larvae wa significantly lower at 1 ppb and higher levels compared with control experiments, and in S. reticulatum a significant decrease in survival began at the 3 ppb level. At 10 ppb Dimilin, no larvae survived to the megalopa stage in either of the two species. The results indicate that early stage larvae of R. harrisii are more sensitive to Dimilin than those of S. reticulatum. When R. harrisii larvae were treated with 10 ppb Dimilin during the intermolt period of each of the 4 zoeal stages, nearly all larvae died during molting to the succeeding stage. First zoeal larvae of R. harrisii exposed to 10 ppb Dimilin at various days during the intermolt period were more sensitive to the compound late than early in the period. It is suggested that Dimilin also may interfere with the formation of the cuticle in crab larvae.     

Planktonic larval duration of one hundred species of Pacific and Atlantic damselfishes (Pomacentridae)

The plankton larval duration for 100 species of Pacific and Atlantic damselfishes was estimated from daily growth increments on the otolith of juvenile fish collected at various localities between July 1987 and September 1988. For newly-settled fishes, larval duration was determined by counting the entire number of increments present on the otolith, while for older juveniles estimates were made by counting the number of increments between the center of the otolith and a mark corresponding to settlement. We document the development of otolith formation during the period when eggs are incubated on the reef and show that daily increments are only accreted after larvae hatch and enter the planktonic phase. The planktonic larval duration for damselfish is shorter and less variable, both between and within species, compared to other groups of reef fishes such as wrasses and surgeonfishes. Larval duration ranged from 12 to 39 d. Average duration between species ranged from 13.1 to 35.2 d. The time spent in the plankton was not significantly correlated with geographic distribution when evaluated among species, however, genera with confined regional distribution have a shorter mean larval life than do widely distributed genera. Size at settlement was positively correlated with time spent in the plankton among species, but a significant correlation between these variables was only evident within one of ten species. The low variance in planktonic larval duration within species indicates that most damselfish are unable to delay metamorphosis following competency. This inability to postpone settlement limits the potential for dispersal, especially when dispersal time between suitable habitats is greater than about 30 d.     

Evolution of reproductive features and larval development in the genus Amphiglena (Polychaeta: Sabellidae)

Larval development in Amphiglena mediterranea is direct in the parental tube, and larvae crawl away as young juveniles with a radiolar crown capable of feeding. The non-feeding larvae of A. nathae are initially brooded in the parental tube, and then have a swimming phase before settling and developing a radiolar crown. Developmental data and reproductive features such as sperm structure are combined with other morphological data for an assessment of the cladistic relationships of the species in the genus Amphiglena. Two species of Laonome are included as part of the ingroup. Six genera, representing a sister group to the Laonome/Amphiglena clade, are used as an outgroup. Two most-parsimonious cladograms were found, and the evolution of reproductive features are discussed. The monophyly of Amphiglena is indicated by features previously identified, as well as two features based on sperm morphology. At present a plesiomorphic larval form cannot be identified for the genus Amphiglena. Received: 16 June 1997 / Accepted: 29 January 1998     

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

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