The use of metabolic ratios for determining the catabolic substrates of a solitary ascidian

In a previous study, Hatcher (1989: Mar. Biol. 102: 445–452) found that variations in CO₂ and O₂ respiration rates in individual marine invertebrates led to RQ (respiratory quotient) values which were variable and often outside theoretical limits. The present study was designed to examine the variability in several excretionbased metabolic ratios which are often used as alternatives for the RQ in qualitative predictions of catabolic substrates. The experimental organism was a solitary ascidian,Herdmania momus (Savigny), collected near Perth, Western Australia, between January and July 1984. Respiration and excretion rates ofH. momus were examined as a function of a progressive nutritional stress, and covariation was examined. Nutritional stress accounted for more of the variation over time in respiration rates (40 to 50%) than in NH ₄ ⁺ excretion rates (20%). Significant net exchanges of dissolved organic compounds were measured. Qualitative predictions of catabolic substrates were based on a comparison of metabolic ratios with theoretical limits. The O:PO ₄ ^3- ratios were lower than the theoretical limits. The values of the O:NH ₄ ⁺ and NH ₄ ⁺ :PO ₄ ^3- ratios were not influenced by nutritional conditions but changed as a function of reproductive condition of the ascidians. Based on the results of this study, it was concluded that metabolic ratios measured on individual ascidians cannot be reliable predictors of catabolic substrates.     

Reproduction in five species of crinoids at Lizard Island,Great Barrier Reef

Reproduction in five species of crinoids [Himerometra bartschi (A. H. Clark), H. robustipinna (P. H. Carpenter), Cenometra bella (Hartlaub), Colobometra perspinosa (P. H. Carpenter), and Oligometra serripinna (P. H. Carpenter)] was studied at Lizard Island, Queensland, over 24 mo, from April 1981 to March 1983 (except for O. serripinna —June 1981 to March 1983). All five species of crinoids were dioecious, although a few instances of synchronous hermaphroditism were noted in H. bartschi, C. perspinosa, and O. serripinna. Gonad morphology and the sequence of developmental stages were similar in each species. Within individuals, gametogenesis was synchronous in gonads both along and between arms, except in some proximal and distal genital pinnules in which gonads never matured. Within a species, there was a degree of synchrony of gametogenesis in a sample, but the level of gametogenic activity varied considerably between years. A high level of continuous reproductive activity was observed in H. bartschi, H. robustipinna, Cenometra bella, and Colobometra perspinosa from about mid-summer to early/mid-winter, while O. serripinna probably had a bimodal reproductive cycle with peaks around February and June. There was a significant 1:1 male:female sex ratio in the populations sampled, and unsexable individuals were significantly smaller (as determined by maximum arm length) than either sexable males or females.     

Response of the predatorRhizophagus grandis to host plant derived chemicals inDendroctonus micans larval frass in wind tunnel experiments (Coleoptera: Rhizophagidae,Scolytidae)

Summary Adults of the predatory beetleRhizophagus grandis are strongly attracted to both adult and larval frass of its specific prey,Dendroctonus micans, in walking bioassays. Spruce bark and resin are relatively unattractive. Solvent extracts of larval frass that were attractive toR. grandis adults in a flight wind tunnel contained a mixture of monoterpenes of host plant origin. A synthetic mixture of these monoterpenes, (+)—-pinene, (–)—-pinene, -phellandrene,dl limonene and 3-carene, was responsible for 70–80% of the activity of the most attractive extract. We suggest that a blend of monoterpenes in frass acts as a kairomone forR. grandis but that attraction to monoterpenes is only the first of a sequence of behavioural responses that ensures successful host location and identification.     

Protein binding of iron in blood plasma of the ascidian Herdmania momus

Blood of the ascidian Herdmania momus var. grandii (Kott, 1952) has typically a low cell haematocrit volume, <1% with a cell density of 12 000 to 15 000 cells mm^-3. Non-dialysable solids in the plasma are 105±10 mg%. Iron-binding in the plasma has been investigated by fractionation of ⁵⁹Fe-labelled plasma, using gel chromatography and electrophoretic procedures. Three fractions, identified as I, II and III in the order of their elution from a G-150 column, bind ⁵⁹Fe in the ratio 60:15:15. The dominant fraction, I, was shown by electrophoresis on polyacrylamide gels to consist of two components in the ratio of ca. 80:20 and with similar electrophoretic mobilities. Dissociation of Fraction I by mercaptoethanol and sodium dodecyl sulfate gave on electrophoresis three components of molecular weights 17 000; 32 000 and 66 000. Fraction 1 contained 10% carbohydrate by weight as determined by the -cysteine/sulphuric acid assay. These data are compared with other iron-binding systems such as transferrin and the siderophores. Phylogenetic aspects of the study are also discussed.     

RQ of benthic marine invertebrates

This study investigated an incubation method which employed simultaneous measurement of CO₂ production and O₂ consumption rates to calculate the RQ (respiratory quotient; CO₂ production rate: O₂ consumption rate) of individual benthic marine invertebrates. Carbon dioxide production rates were calculated from changes in CO₂ concentration determined using seawater pH. O₂ consumption rates were calculated from changes in O₂ concentration with a correction applied for O₂ flux across the air/water interface due to gaseous exchange. Species examined were Triphyllozoon sp. cf. moniliferum (MacGillivray 1860), a bryozoan; Herdmania momus (Savigny), a solitary ascidian; Poneroplax albida (Blainville 1825), a chiton; and Haliotis roei (Gray 1826), an abalone. Six individuals of each were collected on 14 November 1985 from the limestone walls of a cave in a nearshore reef off Marmion, Western Australia. After acclimation for 6 h in experimental conditions, rates of CO₂ production and O₂ consumption were measured. A minimum period of 4 h was required to obtain consistent RQ values for each species. The standard error (SE) of the (calculated) RQ ratio was 14 to 33% of the mean in incubations of 4 h, and less than 14% in incubations of 4 to 12 h. The RQ is commonly used as an indicator of unknown catabolic substrates by comparing it with biochemically determined limits for known substrates. This study provides a strong argument against using the RQ of individual animals to draw any conclusions about catabolic substrates. Unexplained variation in the components of the RQ of an individual, measured over short time periods, and the potential involvement of stored reserves in catabolism, over longer time periods, obscure the relationship between the RQ of individual animals and the ratio's biochemically determined limits.     

Temporal and vertical distribution of two ecologically different calanoid copepods (<Emphasis Type="Italic">Calanoides carinatu</Emphasis>s Krøyer 1849 and <Emphasis Type="Italic">Lucicutia grandis</Emphasis> Giesbrecht 1895) in the deep waters of the central Arabian Sea

The full-depth vertical distributions of males, females and juveniles of Lucicutia grandis and Calanoides carinatus are presented from the surface down to depths of 3,900 m in the central Arabian Sea at the end of the NE monsoon and the February and October inter-monsoonal periods. L. grandis is a steady component and C. carinatus is a temporal resident of the central Arabian Sea oxygen minimum zone. Maximum abundances of C. carinatus and L. grandis juveniles were found at depths around 600 m, whereas the center of the distribution of L. grandis adults was somewhat deeper at 950 m. Both copepods are indicator species which can help to understand the functioning of the mesopelagic Arabian Sea ecosystem.     

Orientation ofRhizophagus grandis (Coleoptera: Rhizophagidae) to oxygenated monoterpenes in a species-specific predator-prey relationship

Summary The predator,Rhizophagus grandis, is linked to its specific prey,Dendroctonus micans, by semiochemical signals regulating oviposition and long range orientation. A mixture of simple oxygenated monoterpenes [(–)—fenchone, (–)—pinocamphone, rac. camphor, terpinene-4-ol, borneol, fenchol and verbenone], identified from the frass ofD. micans has been found to be extremely active in a flight windtunnel as a long—range attractant for the predator. The mixture elicited 84 % of the response to larval frass of the prey. Excluding pinocamphone from the mixture did not influence its activity. Also, changing the absolute configurations of some of the components (fenchol, terpinene-4-ol and borneol) did not influence the predators' response. However, the addition of (–)—-terpineol increased the attractivity of the synthetic blend to almost that of larval frass of the prey. Ecological implications of the identified semiochemicals and their use in pest management are discussed.     

The encrusting spongeHalisarca laxus: population genetics and association with the ascidianPyura spinifera

The encrusting spongeHalisarca laxus forms a seemingly obligate association with the stalked solitary ascidianPyura spinifera. In 1991 we examined spatial variation and short-term temporal variation in this association at three neighbouring sites in southeastern Australia. This sponge dominated the surface of almost all the 500 individual ascidians examined, with mean cover usually exceeding 90%. This pattern was consistent among sites and throughout the year of the study. The domination of a small isolated patch of habitable substratum by a sponge is most unusual, given that they are regarded as relatively poor recruiters. To understand how this association might be maintained, we determined the underlying genotypic diversity of the sponge population using starch-gel electrophoresis.P. spinifera is a clump-forming ascidian and usually occurs in clumps of up to 22 individuals. Electrophoretic surveys, based on six variable allozyme loci, revealed that at a total of five plots within three neighbouring New South Wales populations, single sponge genotypes may cover entire ascidian clumps; although a clump sometimes played host to more than one sponge clone. Allele frequencies (averaged across four loci that appear to conform to Mendelian inheritance) showed little variation among populations (standardised genetic variance,F _ST=0.013). Nevertheless, sponge populations were genotypically diverse, with samples from 63 of 172 individual clumps displaying unique clonal genotypes. Moreover, multi-locus genotypic diversity within all sites approached the level expected for sexual reproduction with random mating. Taken together, these data imply thatH. laxus produces sexually-derived larvae that are at least moderately widelly dispersed. Given the relatively small size of the patches that this sponge inhabits, we also conclude that these larvae are good colonists and good spatial competitors on their ascidian hosts.     

Grazing of phytoplankton by copepods in eastern Antarctic coastal waters

Chlorophyll a, primary productivity and grazing by copepods on phytoplankton were measured in the upper water column during the summer of 1994/1995 at a coastal site near Davis Station, East Antarctica. Chlorophyll a was at a maximum in mid-December, then dropped markedly as the coastal fast ice melted and broke‐out. Phytoplankton biomass increased again from mid‐ to late‐February. Copepods accounted for at least 65% of zooplankton biomass in the water column before sea ice break‐out, whereas larval polychaetes and ctenophores dominated after ice break‐out. Oncaeacurvata was the numerically dominant species throughout the study. The highest grazing rate (8.7 mg C␣m⁻³␣d⁻¹) was recorded on 21 December when O.␣curvata accounted for 64% of the total. Grazing had decreased markedly by 28 December (0.9 mg C m⁻³ d⁻¹); again O. curvata accounted for over 50% of the total ingested. Copepod grazing increased after ice break-out until the last experiment on 20 February (⋍5 mg C␣m⁻³␣d⁻¹). The main species responsible for grazing during this period were O. curvata, Oithonasimilis, Calanoidesacutus and unidentified copepod nauplii. It was estimated that copepods removed between 1 and 5% of primary productivity. Received: 11 October 1996 / Accepted: 22 October 1996     

Storage of exogenous sperm by the compound ascidianDiplosoma listerianum

Experimental evidence is presented for the storage of exogenous sperm by zooids of the simultaneously hermaphroditic didemnid ascidianDiplosoma listerianum. Cross-fertilized zygotes can be produced up to 1 mo after a ramet is placed in reproductive isolation. Previously published observations, considered in the light of the present data, suggest that the lumen of the ovary is the site of sperm storage. This is believed to be the first report of post-transmission storage of sperm by an ascidian. Existing records of storage of exogenous sperm in other taxa rarely involve aquatic animals in which spermatozoa are transmitted between mates singly through the surrounding water, as inD. listerianum. The potential significance of sperm storage in the mating systems of sessile aquatic invertebrates is discussed.     

Population and reproductive biology of two sibling amphipod species from ascidians and sponges

 The population dynamics and reproductive biology of an ascidian- and a sponge-dwelling amphipod were examined. The two undescribed amphipod species, Leucothoe“ascidicola” and L.“spongicola”, are closely related to each other, and occur in ascidians and sponges, respectively, along the Florida Atlantic coast. L. “ascidicola” was abundant in solitary ascidians during fall 1997, disappeared during spring/early summer, and became abundant again in September 1998. During the time when L. “ascidicola” were absent from their hosts, a copepod became a frequent inhabitant of the ascidians but disappeared again when L.“ascidicola” returned to the ascidians in September 1998. The numbers of L.“spongicola” in sponges increased substantially during spring, when high reproductive activity was observed. Following this reproductive peak, both adult and juvenile amphipods apparently left the sponges, and during the summer amphipod numbers in the sponges were very low. Another small amphipod species, which often co-occurred with L.“spongicola”, showed less seasonal variation and was found in sponges throughout the whole study period. The percentage of ovigerous females per host unit was usually lower in the ascidian-dwelling than in the sponge-dwelling amphipods. In solitary ascidians, L.“ascidicola” amphipods usually occurred in groups of several adults, yet there never was more than one ovigerous female per ascidian. In contrast, several ovigerous L.“spongicola” females were found to cohabit in the same spongocoel. This suggests that intrasexual aggression may be stronger among reproductive amphipod females in the ascidians than in sponges. The size distributions of juvenile cohorts indicate that juvenile L.“ascidicola” remain for relatively long time periods in the parental ascidian, where they may reach sexual maturity. In contrast, in L.“spongicola”, only cohorts of very small juveniles could be identified, indicating that juveniles disperse shortly after emerging from the female's brood pouch. It is concluded that extended parental care is of very short duration or does not occur in the sponge-dwelling amphipod L. “spongicola”, possibly because fast-growing sponges with a highly branched spongocoel system do not allow long-lasting coexistence of parent-offspring groups. In contrast, the discrete character of the solitary ascidians may enhance the potential for exclusion of other species, resource monopolization by reproductive females, and furthermore for long-lasting extended parental care in the ascidian-dwelling amphipod. Groups of single parents together with cohorts of large juveniles are reported in the literature for amphipods and isopods from brachiopods, bivalves and ascidians, suggesting that these discrete biotic microhabitats may favor the evolution of extended parental care in peracarid crustaceans. Received: 30 July 1999 / Accepted: 8 May 2000     

Light-enhanced growth of the ascidian Didemnum molle/Prochloron sp. symbiosis

To obtain a direct measurement of the importance of Prochloron sp. to the ascidian Didemnum molle, ascidian colonies from Lizard Island Lagoon, Great Barrier Reef, were grown for 9 d at 0, 10, 40 and 100% sunlight in situ using unidirectional flow chambers. Growth (wet weight) was enhanced up to 40% of full sunlight, at which point growth appears to have been light-saturated. Colonies in 10 and 40% sunlight responded by (1) climbing up the sides of the growth chambers, and (2) flattening out to a more encrusting morphology; also (3) the chlorophyll content of three colonies in zero sunlight decreased by >80%, yet the ascidians remained healthy and did not lose weight. These data show that although the symbiosis may not be obligatory for D. molle, the ascidian's growth is enhanced by Prochloron sp., and the morphology of the ascidian colony is affected by its photobiology.Contribution No. 344 of the Australian Institute of Marine ScienceCommunicated by G. F. Humphrey, Sydney     

Heat tolerance,growth and regeneration in three North Sea bryozoans exposed to different constant temperatures

Three species of bryozoans—Membranipora membranacea (L.), Electra pilosa (L.) and Conopeum reticulum (L.) — are capable of acclimating to elevated temperatures, above the normal range experienced in nature, when exposed to a gradual increase in ambient temperature. Conspicuous differences in LD 50 values, as a consequence of acclimation, occur between representatives of the same species acclimated and grown at constant temperatures in the laboratory. The tolerance range of these species is influenced by their thermal history in the laboratory. While increased ambient temperatures accelerate growth rate, final colony size attained after prolonged exposure declines at higher temperatures. The size of zooecia attained is inversely proportional to the test temperature. Colonies of E. pilosa maintained at 22°C develop erect branches. Hence, it is probable that E. pilosa forma erecta is only a growth form of normally encrusting colonies of E. pilosa. Temperature affects rate of regeneration.     

Reproductive biology of the holothurians from the major communities of the New Caledonian Lagoon

The reproductive biology of nine species of aspidochirote holothurians from the major communities of the New Caledonian lagoon has been analysed. The data were obtained from monthly sampling from 1978 to 1982 of their populations on reef slopes and passes for Holothuria nobilis, H. fuscogilva, H. fuscopunctata and Thelenota ananas, in the inner lagoon for Stichopus variegatus, H. scabra and H. scabra versicolor and on reef flats for Actinopyga mauritiana, A. echinites and H. atra. The morphology and anatomy of the gonads and their annual cycle are described. The reproductive cycles, composed of the five phases of gonad growth, maturing, spawning, post-spawning and resting, were determined from the changes in the sexual stages, the gonad indices and the percentage of indeterminate sex. Three main reproductive patterns are recognized, annual cycles with spawning occurring during the warm season or during the cool season and a semiannual cycle. Other population parameters, such as the size of the mature oocytes, the mean size of the individuals at first sexual maturity, and absolute and relative fecundities were calculated and related to the mean size of the species, the taxonomy and the ecology. The reproductive strategies of the species show a gradient that is probably related to the stability of the biotopes, with smaller species living in the more intrable outer reef flats and larger species in deeper lagoon floors.     

Larval depletion by ascidians has little effect on settlement of epifauna

Ascidian densities were manipulated while controlling for negative effects of biodeposition and space preemption to examine the effects of ascidian filter-feeding on larval recruitment in St. Joseph Bay and near Turkey Point, Florida (Northern Gulf of Mexico, Florida, USA). Using three different experimental designs during 1984 and 1985, recruitment near living Styela plicata or Molgula occidentalis was as high as recruitment near ascidian models. Disruption of flow by ascidian bodies had little effect on settlement. Predation rates by ascidians on larvae in six phyla were high in laboratory experiments. The field effects of larval depletion by solitary ascidians are apparently obscured by other factors influencing the abundance of recruiting larvae. Consumption of larvae in the laboratory cannot be used to assume significant inhibitory effects in the field.     

Spermatozoan ultrastructure and spermatogenesis in aplousobranch ascidians,with some phylogenetic considerations

Spermatozoa and some stages of spermatogenesis were studied for four species of aplousobranch ascidians. Spermatozoa of Clavelina lepadiformis (Müller) (family Clavelinidae) are plesiomorphous in that they have apical acrosomal vesicles and a moderately elongated, cylindrical nucleus. The elongated mitochondrion is twisted ea. 11/2 times around the nucleus. In their ultrastructural morphology, C. lepadiformis sperm conform in some respects to the least-derived ascidian sperm, those of the phlebobranch ascidian Ciona intestinalis L.; however, the sperm of Clavelina lepadiformis have two apomorphies not shared with those of Ciona sp.: (1) the mitochondrion of Clavelina lepadiformis is long and spiralled along the entire nucleus rather than being comparatively compact and not at all helical; (2) the mitochondrial cristae are elongated parallel to the long axis of the nucleus, whereas in Ciona sp. sperm the cristae are unmodified. In Distaplia sp., Aplidium sp. and Synoicum pulmonaria (Ellis and Solander) the spermatozoa are more derived and consist of a proximal cylindrical and a distal corkscrew-like part. The mitochondrion in Distaplia sp. and Aplidium sp. contains electron-dense material and extends in a long thread around the nucleus. In S. pulmonaria the mitochondrion surrounds the anterior part of the nucleus in mature spermatozoa, and an elongated, dense structure displaying fine striation is enclosed in the mitochondrion in late spermatids. The sperm ultrastructural morphology observed in this study is consistent with the majority view that clavelinids are closest to the ancestral ascidian but is also consistent with other conclusions, particularly that the Cionidae are closest to the stem ascidian.     

Localization of granulatimide alkaloids in the tissues of the ascidian <Emphasis Type="Italic">Didemnum</Emphasis><Emphasis Type="Italic">granulatum</Emphasis>

Ascidians are a notable source of nitrogen-bearing secondary metabolites with a wide range of biological activities. Although many biologically active compounds have been isolated from ascidians, it is often unclear whether the animal or associated microbial symbionts such as bacteria or fungi are the true biosynthetic source of the metabolites. We have addressed the question of the biosynthetic source of the alkaloids granulatimide and isogranulatimides by localizing these compounds within the ascidian. In this work, we demonstrate that granulatimide is stored in Didemnum granulatum tunic bladder cells. Analysis by confocal fluorescence microscopy at the granulatimide emission range indicated the presence of fluorescent cells as highly vacuolated cells found dispersed in the ascidian tunic. Since this is the most exposed ascidian tissue, it is possible that these alkaloids may have a protective role, either as sunscreens and/or as feeding deterrents.     

The reproductive ecology of the invasive ascidian, <Emphasis Type="Italic">Styela clava,</Emphasis> in Auckland Harbour,New Zealand

The ascidian Styela clava, native to the north-west Pacific, is an invasive species affecting New Zealand’s marine ecosystems, biodiversity and aquaculture operations. To provide detailed information on the reproductive biology of S. clava in New Zealand for post-border biosecurity management, long-term seasonal patterns of gametogenesis were determined from May 2006 to May 2008 in Auckland’s Waitemata Harbour (36°49′20″S, 174°45′85″E). Of particular interest was whether the critical 15°C threshold spawning temperature for reproduction observed in the Northern Hemisphere applied here to the first Southern Hemisphere study. S. clava gametogenesis followed a regular seasonal cycle with ripe gametes appearing as early as September and persisting to June; this time frame corresponds to the period when sea surface temperatures in the region first reach 15°C and with spawning occurring mainly during late summer to early autumn. From photoperiod manipulation, it was determined that spawning occurred at approximately 18:20. The extended reproductive period and a short generation time in the Waitemata Harbour provides a lengthy opportunity for S. clava to spread. Findings are discussed in relation to S. clava’s post-border management.     

Symbiotic zooxanthellae enhance boring and growth rates of the tropical sponge Anthosigmella varians forma varians

Several species of boring sponges harbor symbiotic zooxanthellae, and it is believed that the symbiont enhances boring activity of host sponges. This hypothesis was tested using manipulative field experiments to assess the effect of intracellular zooxanthella populations on boring rates of the tropical sponge Anthosigmella varians forma varians. Portions of sponge were attached to 60 calcium carbonate blocks of known weight. Three sets of 10 blocks were grown at high light levels and three sets of 10 blocks were grown at low light levels for 105 d in the Florida Keys, Florida, USA. Boring rates, growth rates (lateral growth and within-substratum tissue penetration), and zooxanthella populations were measured at the end of the experiment. Absolute rates of boring and growth of A. varians forma varians were significantly greater when zooxanthella densities were higher. Boring rate and tissue penetration related to final surface area of sponge attachment was also enhanced when zooxanthella densities were higher, suggesting that the symbiont plays a physiological role in the decalcification process. This is in contrast to the role that zooxanthellae play in coral hosts. Based on the results of this study, it appears that the presence of zooxanthellar symbionts has important ecological and life-history consequences for host sponges. Ability to laterally overgrow competitors will be correlated with the size and activity of zooxanthella populations. In addition, the fitness of host sponges will be enhanced by algal symbionts, since greater penetration within substrata will result in an increase in production of tissue that can be converted into storage, feeding and reproductive functions.