Association of bacteria with larvae of the gutless protobranch bivalve Solemya reidi (Cryptodonta: Solemyidae)

Rod-shaped bacteria were consistently observed by transmission electron microscopy in the locomotory test of larvae and in the perivisceral cavity of post-larvae of Solemya reidi, a gutless protobranch bivalve known to possess intracellular chemoautotrophic bacterial symbionts in the adult gill. Bacteria develop within granular vesicles in the larval test, where they either remain to be ingested at metamorphosis, or are released into the space separating the test and embryo, to be subsequently ingested through the larval mouth. In either case, bacteria lie within the perivisceral cavity following metamorphosis. Bacteria were not seen either in or on gametes or in gills of juveniles. It is hypothesized that these bacteria represent a transmission stage of the gill symbionts present in adult S. reidi and are not evident in gametes or gills of juveniles due to cryptic packaging within granular vesicles. Perpetuation of this symbiosis would therefore be assured through vertical transmission, as is typical of other marine invertebrate-bacteria endosymbioses.Harbor Branch Oceanographic Institution Contribution No. 602     

Development of the pericalymma larva of Solemya reidi (Bivalvia: Cryptodonta: Solemyidae) as revealed by light and electron microscopy

Solemya reidi Bernard 1980 is a gutless protobranch bivalve known to possess intracellular chemoautotrophic bacterial symbionts in its gill. A light and electron microscope study on the embryology and larval development of S. reidi provides data for the bivalve Subclass Cryptodonta. S. reidi spontaneously spawned large eggs (271 m in diameter), which developed within individual gelatious egg capsules. The first several cleavages were equal and a distinct molluscan cross was formed at the animal pole of the embryo, features previously unreported in bivalve development. Lecithotrophic pericalymma larvae (similar to the larvae of paleotaxodont protobranch bivalves and aplacophoran molluscs) hatched at 18 to 24 h and remained in the water column for a further 5 d at 10°C. At hatching, larvae measured from 360 to 440 m in length and from 225 to 265 m in cross-sectional diameter. Definitive adult structures developed within an epithelial locomotory test entirely covered with compound cilia. The test histolysed at metamorphosis and was ingested throught the mouth into the perivisceral cavity. Length and height of the shell following metamorphosis was 433 m (±42 m, n=16) and 282 m (± 29 m, n=13), respectively. Primary data and data from the literature show that the type of larval development in both paleotaxodont and cryptodont bivalves cannot be reliably estimated from egg or prodissoconch sizes.     

Translocation of fixed carbon from symbiotic bacteria to host tissues in the gutless bivalve Solemya reidi

Naturally-occurring lipophilic inducers of larval settlement and metamorphosis wer isolated and identified for Phragmatopoma californica, a gregarious tube worm from southern California. Organic solvent extraction of the sand/organic cement matrix of tubes diminished the inducing capacity of the tube matrix. The inducing capacity was restricted to a single, highly active, HPLC-purified fraction of the organic solvent extract. Chemical analysis of this fraction revealed a mixture of free fatty acids (FFAs), dominated by eicosapentaenoic acid (20:5, 20%), palmitic acid (16:0, 14%) and palmitoleic acid (16:1, 12%). In assays of the nine FFAs that each contributed 3% or more to the active fraction, only 16:1, 18:2, 20:4 and 20:5 induced larval settlement and metamorphosis, while the others were ineffective. The larval response was contact-dependent, highly specific, and concentration-dependent, with a significant response to 16:1 and 20:4 at as low as 10 g FFA spread onto 1 g of sand (surface area 36 cm²). Active FFAs were extracted at approximately 14 g g^-1 sand from the tube matrix, although the levels encountered by larvae in nature are believed to be higher.     

Oxygen transport by the hemocyanin of the protobranch mollusc Solemya reidi

Oxygen transport by the hemocyanin of the protobranch mollusc Solemya reidi Bernard was studied in native hemolymph samples. Clams were collected from two different reducing environments, beneath log booms in Alberni Inlet, British Columbia, Canada, and from the sewage effulent in Santa Monica Bay, California, USA. The hemocyanin concentration in a pooled hemolymph sample (n = 10 individuals) was 33.5 mg ml⁻¹. The mean hemolymph pH of five Alberni clams maintained for 3 wk in mud was 7.96 ± 0.06. No significant variation in hemocyanin oxygen-affinity or cooperativity was found for hemocyanin in whole hemolymph samples from these five individually studied clams. There was a significant difference only at 15 °C in the oxygen affinity of hemocyanin in pooled whole-hemolymph samples from S. reidi collected from Alberni Inlet compared with clams collected from Santa Monica Bay. Little effect of temperature on hemocyanin oxygen-affinity was found for temperatures below 20 °C; above 20 °C, the oxygen affinity was reduced for clams from both sites. Temperature and pH had no apparent affect on hemocyanin cooperativity. Moderate Bohr shifts were found at all temperatures examined. The presence of physiologically relevant concentrations of thiosulphate in hemocyanin samples resulted in a decrease in hemocyanin oxygen-affinity, opposite to the effect on hemocyanin found for the hydrothermal vent crab Byth-ograea thermydron, but thiosulphate had no effect on hemocyanin cooperativity. Received: 2 September 1997 / Accepted: 6 February 1998     

Larval period and its influence on post-larval life history: comparison of lecithotrophy and facultative planktotrophy in the aeolid nudibranch Phestilla sibogae

The variable duration of the pelagic phase of metamorphically competent larvae of benthic marine invertebrates is set by an interaction between environmental factors and larval traits that together influence the chance that a larva will encounter and respond to a suitable settlement site. In the Hawaiian aeolid nudibranch Phestilla sibogae Bergh, an extended competent larval phase resulted in a cascade of negative effects on larval and post-larval life-history traits. When raised as fed (i.e., facultatively planktotrophic) larvae, an extended larval period resulted in lower larval survival, slightly lower metamorphic success, and delayed reproduction. When raised as unfed (i.e., lecithotrophic) larvae, an extended larval period resulted in lower larval and post-larval weights, survival, metamorphic success, and reproductive output, and also resulted in a longer juvenile period and delayed reproduction. The chance nature of locating a settlement site generally spreads these negative effects over all larvae of a cohort, and so balances the relative fitness of the genetic lineages within a population.     

Localization of RubisCO and sulfur in endosymbiotic bacteria of the gutless marine oligochaete Inanidrilus leukodermatus (Annelida)

 The chemoautotrophic potential of the two co-occurring larger and smaller bacterial endosymbionts of the gutless marine oligochaete Inanidrilus leukodermatus was determined using immunocytochemistry. An antibody directed against the Form I of ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO), the key CO₂-fixing enzyme of the Calvin–Benson cycle, consistently labeled the larger symbionts. Electron microscopic spectroscopy showed that the larger symbionts contained sulfur in intracellular globules and to a lesser degree in the cytoplasm. The presence of RubisCO and sulfur indicates that the larger endosymbionts of I. leukodermatus are chemoautotrophic sulfur-oxidizing bacteria. In contrast, no RubisCO or sulfur was detected in the smaller endosymbionts of this host. Received: 28 September 1999 / Accepted: 29 May 2000     

Larval development in the Antarctic nemertean Parborlasia corrugatus (Heteronemertea: Lineidae)

Embryonic and larval development were followed from fertilisation to settlement in the Antarctic heteronemertean Parborlasia corrugatus (McIntosh, 1876). The first cleavage occurred 10 to 15 h after fertilisation, and the second at 17 h. Larvae hatched at the gastrula stage, between 170 and 200 h post-fertilisation, and were 150 m in diameter. Early larval stages aggregated in dense groups near the surface of incubation vessels and were positively phototactic. Early pilidium larvae were recognisable 435 h post-fertilisation. They were 155×152 m in size, and possessed a complete apical tuft of cilia and a full marginal band of locomotory cilia. At this stage, the gust was visible through the body wall, and the mouth was open and was 40 m in diameter. Late pilidia, 222×193 m in size, were helmet-shaped. They had an apical tuft over 100 m long, and possessed a lobed marginal band of locomotory cilia. Pilidia were observed aggregating close to the bottom of incubation vessels 1200 to 1350 h (50 to 56 d) after fertilisation, and this was interpreted as settlement behaviour. At this stage, the apical tuft had been lost and they were highly contractile, being capable of compressing their bodies. However, neither developing juveniles within the larval envelope nor hatched juveniles were observed. Pilidia consumed the microalgae Tetraselmis suecica, Thalassiosira pseudonana and Isochrysis galbana. They also fed on particulate organic material < 1 m in size, as shown by the presence of material in the guts of larvae offered filtered extracts of algal cultures. There was some indication that larvae could use dissolved organic material, since pilidia held in seawater with organic material removed did not survive as long as those in filtered seawater or in filtered water with added amino acids. However, the only larvae to exhibit settlement behaviour in the feeding experiments were those offered Tetraselmis succica and Thalassiosira pseudonana, and these required a longer development time to reach this stage than pilidia in the standard cultures, where a mixed algal diet was offered.     

Larval development of Euphausia nana (Crustacea: Euphausiacea)

The larvae of Euphausia nana Brinton in Sagami Bay and Suruga Bay, Central Japan, are described and illustrated. The furcilia stage is separated into six stages on the basis of the form of the pleopods and the number of terminal telson spines. Furcilia I, no pleopod or non-setose pleopods and seven terminal telson spines. Furcilia II, nonsetose and setose pleopods and seven terminal telson spines. Furcilia III, five pairs of setose pleopods and seven terminal telson spines. Furcilia IV, five pairs of setose pleopods and five terminal telson spines. Furcilia V, five pairs of setose pleopods and three terminal telson spines. Furcilia VI, five pairs of setose pleopods and one terminal telson spine. Segmentation of the antennal endopod and mandible palp occurred at the furcilia V stage. The dominant forms of early furcilia larvae of E. nana differed during different seasons of the year. The total lengths of calyptopis III and furcilia I to VI larvae also showed seasonal changes; they were largest in May and smallest from September to December.     

Larval settlement and metamorphosis in the sand crab Emerita talpoida (Crustacea: Decapoda: Anomura)

The effects of various potential cues on the survival and time to metamorphosis of larvae of the sand crab Emerita talpoida (Say) were examined. Zoeal duration, which ranged from 25 to 43 d, was not correlated with subsequent megalopal survival but had a weak, negative correlation with the duration of the megalopal stage. Neither food, sand nor water from containers with adults directly affected megalopal survival or duration. In a second experiment, the presence of sand both increased the probability of survival and shortened the duration of late zoeal stages. This is the first experimental evidence for delayed zoeal metamorphosis in a decapod. Settlement of E. talpoida appears to occur primarily during the zoeal phase, not at the megalopal stage.     

Reproduction,larval development and post-larval growth ofNaineris laevigata (Polychaeta,Orbiniidae) in the Mediterranean Sea

To investigate the life-history adaptations of a brackish-water population ofNaineris laevigata Grube, a population from Acquatina lagoon near Lecce (Italy) was studied. Benthos and plankton samples were collected monthly over a period of ca. 2 yr (1989–1990). Observations of gametogenesis indicate that oogenesis is intraovaric, probably with autosynthetic yolk formation. Oogenesis takes about 7 mo, from November to April–May, for both proliferative and vitellogenetic phases. Reproduction is synchronized within the population. Spawning has also been obtained under laboratory conditions where larval development and post-larval growth were studied. Eggs measured 240 µm in diameter and were spawned within a gelatinous mass. The lecithotrophic pelagic stage was very short. Reproductive effort was high, probably because of high food availability and low level of competition. One of the most interesting findings was the probable presence of a resting stage in the life-cycle. The adaptive significance of enclosing the eggs in gelatiuous masses and of reducing the duration of the pelagic stage is discussed.     

Larval reproduction in the ctenophore Mnemiopsis mccradyi (order Lobata)

Dissogony was observed in normal and experimentally manipulated individuals of Mnemiopsis mccradyi in February–April, 1985. The normal oral-aboral length at which sexual reproduction begins in M. mccradyi is 30±5 mm; however, some larvae, of 1.5 to 2.8 mm (oralaboral length), produce viable gametes. During the present study, not all individuals from a particular spawning which were kept in the same conditions became reproductive at an early age. The age at which larval reproduction began varied between individuals. The length of time over which individuals spawned varied considerably — some spawned only once, while others spawned repeatedly. Only the gonads located under the adesophageal ctene rows (and not those associated with the adtentacular rows) produced gametes during the early reproductive period. After larvae stopped spawning, the gonads regressed and the larvae grew to large lobate stages. Individuals which became sexually reproductive as larvae also became sexually reproductive during lobate stages.     

Larval distribution and reproductive ecology of Thysanoessa macrura (Crustacea: Euphausiacea) in the Scotia Sea

In the Scotia Sea, the larvae of Thysanoessa macrura G.O. Sars, 1885 were scarce in spring (September 27 to October 6, 1971) and mainly present at the Calyptopis I stage, while in summer (December 17, 1974 to January 3, 1975) they were much more abundant and much further developed (up to Furcilia VI). The spawning season commences as early as September and, as indicated by the presence of numerous nauplii and metanauplii in summer, is rather long. T. macrura begins to spawn much earlier in West-Wind-Drift waters than in waters of the Weddell Sea. It starts to spawn simultaneously with Euphausia frigida and E. triacantha, but significantly earlier than E. superba. These timings of the breeding process are related to differential biogeographical characteristics of these species. Plankton collections made at depths of 500 or 1000 m have, for T. macrura, revealed a developmental ascent analogous to that of E. superba. Ontogenetical deep-water migrations are, consequently, characteristic not only for E. superba, but also for other Antarctic euphausiids (in particular E. frigida, but possibly not for inshore E. crystallorophias) and, as correctly postulated by several other investigators, for all oceanic euphausiid species.     

Larval feeding by adult bumble bee workers (Hymenoptera: Apidae)

Summary Laboratory studies on colonies of Bombus terrestris (L.) and B. terricola Kby. showed that while the overall rate of larval feeding is highly correlated with total larval biomass, feeding of individual larvae is only weakly regulated. Nevertheless, the temporal distributions of inspections and feedings to larvae by adult nurse bees did provide evidence for a feedback mechanism.The behavior of individual workers engaged in larval feeding is highly contagious through time. A laboratory colony of B. terrestris compensated for the effect of experimental removal of half the worker population by increases in the feeding rate of individual workers. The results appear to be consistent with recent suggestions that overall colony performance is a mass effect resulting from the partially stochastic operations of its individual components.     

Larval development of Pilumnoides perlatus (Brachyura: Xanthidae) under laboratory conditions

Larvae of the xanthid crab Pilumnoides perlatus (Poeppig, 1836) have been reared in the laboratory at 3 different temperatures (10.2°, 15° and 20°C) from hatching to megalopa stage. The 5 zoea stages and the megalopa, as well as the setation of the functional appendages are described and illustrated. The main characteristics useful to differentiate the larval stages of P. perlatus from those of Homalaspis plana, the other Chilean species of the same family so far reared, are discussed. Data on duration of zoea development, length of moulting intervals, and mortality at the 3 test temperatures are also given.This study was partially supported by the Chilean National Commission for Scientific and Technological Research (CONICYT).     

Larval development of Ocypode quadrata (Brachyura: Crustacea) under laboratory conditions

The larvae of Ocypode quadrata (Fabricius) have been reared in the laboratory, from hatching to megalopa stage, at 35 S, 25°C. The five zoeal stages and the megalopa are described, including functional appendages of each stage. On the basis of morphological characteristics, the first zoeal and megalopa stages of O. quadrata can be distinguished from similar stages of closely related Ocypodinae. At 25°C, the megalopa appeared in a minimum of 34 days following hatching.     

Strobilation,budding and initiation of scyphistoma morphogenesis in the rhizostome Cassiopea andromeda (Cnidaria: Scyphozoa)

Scyphistomae of Cassiopea andromeda Forskål, 1775 containing symbiontic zooxanthellae did not develop medusae at a constant temperature of 20°C, but monodisc strobilation was initiated after transfer of the polyps to 24°C. After release of the ephyrae and regeneration of the hypostome and tentacular region, the recovered polyps either produced vegetative buds or entered a new strobilation phase. Formation of motile, planula-like buds was not found to be indicative of unfavourable environmental conditions. Intensity of budding was positively correlated with available food and with increase of temperature. Budding was negatively correlated with the number of polyps maintained per dish and with the conditioning of the sea water. Under optimal feeding and temperature conditions, polyps could simultaneously produce chains of buds at 2 to 4 budding regions. Settlement and development of buds into scyphistomae was suppressed in pasteurized sea water and in pasteurized sea water containing antibiotics, but polyps developed from buds in the presence of algal material taken from the aquarium, debris or egg shells of Artemia salina, or on glass slides which had been incubated in used A. salina culture medium. Several species of marine bacteria were detected after staining these slides. One, a Gramnegative coccoid rod, which was identified as a nonpathogenic Vibrio species, was isolated, cultivated as a pure strain, and was proved to induce the development of C. andromeda buds into polyps. Millipore filter-plates coated with Vibrio sp. cells grown in suspension culture were ineffectual, but diluted filtrate initiated polyp morphogenesis. The inducing factor is obviously not a constituent of the bacterial cell surface, but is a product of growing Vibrio sp. cells released into the medium. This product was found to be relatively heat-stable and dialyzable. As to the basic mechanism involved in the induction of polyp formation, it is suggested that the inducing factor (s) acts bimodally by inducing pedal disc development and by eliminating a head inhibitor originating from the basal end of the bud. The life history, and various aspects of medusa-formation and of vegetative reproduction in scyphozoans are reviewed and discussed with particular reference to rhizostome species. Special attention has been paid to some reports of larval metamorphosis controlled by marine bacteria.     

Larval biology of six species of the genus Conus (Gastropoda: Toxoglossa) in Hawaii,USA

Among the Hawaiian species in the genus Conus, larval life histories range from long-term planktotrophy to nonpelagic lecithotrophy. Prehatching developmental time, hatching size, pelagic period and total prejuvenile developmental time are all significantly correlated with egg size, while settling size is correlated with none of these larval characteristics. Therefore, larval life history patterns in the genus Conus appear to be determined by 2 sets of selective forces: the first influencing egg size, and the second stipulating settling size. As egg sizes become larger, hatching sizes and prehatching developmental time increase while pelagic periods and total prejuvenile developmental times decrease. Settling size appears to be related to water depth, but food requirements of newly metamorphosed juveniles may also be important.