Molecular characterization of symbiotic algae (Symbiodinium spp.) in soritid foraminifera (Sorites orbiculus) and a scleractinian coral (Orbicella annularis) from St John,US Virgin Islands

The exchange of Symbiodinium symbionts among scleractinian and soritid hosts could facilitate acclimatization to changing conditions by establishing novel symbiotic unions better tuned to prevailing conditions. In this study, we compare the communities of Symbiodinium spp. in neighboring populations of Orbicella annularis and Sorites orbiculus from St. John, US Virgin Islands, using operational taxonomic unit (OTU) clustering of cloned internal transcribed spacer 2 (ITS-2) rDNA sequences. We tested for partitioning of Symbiodinium OTUs by host and depth within and between two sites to explore the potential for symbiont exchange between hosts and light-dependent microhabitat specialization. An apparent lack of overlap in Symbiodinium communities (13 OTUs representing 7 clades) hosted by O. annularis and S. orbiculus suggests that exchange among these hosts does not occur. A low number of novel clade G ITS-2 sequences were found in one O. annularis and one S. orbiculus. A phylogenetic analysis of these sequences revealed them to be sub-clade G2 Symbiodinium, which are most commonly hosted by excavating clionid sponges. A permutational MANOVA revealed within host differences in the partitioning of Symbiodinium OTUs by site but not depth. This finding highlights the potential roles of either dissimilar environmental conditions between sites, or at least partial separation between populations, in determining the types of Symbiodinium contained in different hosts on a spatial scale of a few kilometers.     

Localization and biochemical characterization of pharyngeal protease in the polychaetous annelid Glycera convoluta

In Glycera convoluta Keferstein, pharyngeal protease is secreted by the glandular epithelial cells, as clearly demonstrated by microscopic observation of the digestion of a prestained gelatin film by fresh tissue sections of the organ. The enzyme, extracted by affinity chromatography on Sepharose-?-aminocaproïc-D-tryptophan, acts on substrates hydrolyzable by α-chymotrypsin such as N-benzoyl-L-tyrosine ethyl ester and carboxyl-propionyl-phenylalanine-p-nitroanilide, and can be considered as a chymotrypsin-like enzyme.     

Exceptionally high concentrations of the insect repellent N,N-diethyl-m-toluamide (DEET) in surface waters from Jakarta,Indonesia

Jakarta is a booming coastal megacity in Indonesia with over 10 million inhabitants. The rivers flowing through the city district receive enormous amounts of untreated wastewaters from households and industries and discharge high pollutant loads into Jakarta Bay. Applying a screening approach for the identification of characteristic site-specific and harmful organic contaminants, we frequently found the insect repellent N,N-diethyl-m-toluamide (DEET) in river water and seawater samples from Jakarta. The compound was previously reported as persistent aquatic contaminant in industrial countries, and we present here the first data set from a tropical megacity. Concentrations in river water and seawater from Jakarta were exceptionally high, up to 24,000 ng L^?1, and exceeded by far all published concentrations in surface waters worldwide. We explained this with massive usage of the compound, lack of wastewater treatment and low average river flow as compared to rivers in other tropical megacities. The usage and properties of DEET indicate its suitability as molecular marker of municipal wastewaters. Such markers are useful to trace emissions from specific pollution sources in aquatic systems as a basis for the investigation of related impacts. We show here that DEET is in particular useful to trace the long-range distribution of municipal wastewaters in tropical freshwater and coastal systems. This application is of great value for tracing such inputs in tropical coastal habitats which are sensitive to changing water quality like coral reefs. This assists to uncover whether specific conditions in these systems could be related to pollutant inputs from land.     

Hydrolytic enzymes in the two North Sea ctenophores Pleurobrachia pileus and Beroe gracilis

Hydrolytic enzyme activities were determined in tissue extracts of the two North Sea ctenophores Pleurobrachia pileus and Beroe gracilis. The following activities were measured in field samples and characterized according to their mean specific activities and their pH optima: protease, aminopeptidase, amylase, N-acetylglucosaminidase, -glucosidase, chitinase, carboxylesterase, phosphatase. Many of the enzyme activities are correlated, indicating their control by the same factors (i.e. nutritional conditions). Differences found between the phosphatase activities and the apparent lack of the chitinolytic system in B. gracilis reflect the differences in body composition and food specialization in both ctenophores, respectively. The low amylase and -glucosidase activities in both ctenophores agree with the low carbohydrate content of their prey. In spite of high carboxylesterase activities, no lipolytic activity was detected in either of the ctenophores. In a laboratory feeding experiment with P. pileus, the enzyme activities increased but did not reach the activities found in the field samples. In B. gracilis, seasonal variations of some enzyme activities are discussed in relation to the seasonal abundance of its prey, P. pileus.     

Dimethyl disulfide and dimethyl trisulfide: so similar yet so different in evoking biological responses in saprophilous flies

Dimethyl disulfide (DMDS) and dimethyl trisulfide (DMTS) are used by saprophilous insects to locate breeding sites (decaying organic matter), and by brood-site deceptive flowers to attract such insects. However, little is known about the relative importance of these two compounds in eliciting electrophysiological and behavioral responses in the insects. Here, we compared the relative attractiveness of DMDS and DMTS to saprophilous flies in field choice experiments and tested whether potential differences in field responses can be explained by differences in electrophysiological antennal responses to these compounds. Field experiments revealed that the attractiveness of a mixture of these compounds is due to DMTS alone. This result was confirmed by electroantennographic recordings in which flies of four tested species of Calliphoridae (Lucilia sericata, L. caesar, Calliphora vicina, Protocalliphora azurea) and one Muscidae (Musca domestica) respond clearly to DMTS, but not to DMDS. In house flies, however, DMTS elicited electrophysiological responses only, not reflected in behavioral assays. Despite the fact that DMTS and DMDS exhibit similar chemical structures, both the electroantennographic and field responses from saprophilous flies to these two compounds strongly differed. Our study suggests that oligosulfide-responsive saprophilous flies rely on DMTS and not DMDS for finding appropriate breeding sites and that DMTS and not DMDS could act as a key mediator for pollinator attraction in brood-site deceptive plants.     

Morphological correlates of intraspecific grafting reactions in the marine demosponge Callyspongia diffusa

There are profound differences in the ways that the sponge Callyspongia diffusa reacts to self versus non-self tissue contact. Syngeneic or isogeneic contacts result in rapid tissue fusion and persistent maintenance of normal sponge structure. However, allogeneic contacts result in immune-type rejection reactions leading to tissue death at the interface. Using grafted sponge tissues, both types of response were subject to fine-structural analysis by scanning electron microscopy. Isografts showed no overt cellular reactions other than compatible fusion, whereas sponge tissues at allograft interfaces showed incompatibility reactions. The major steps in the histopathologic sequence of events in allograft reactions may be summarized as follows: (1) immediate allorecognition occurs accompanied by microscopic cellular rearrangements; (2) macroscopic tissue bridges form across the graft interface; (3) massive cellular infiltration towards graft sites, with cells often passing into a graft partner through the tissue bridges; (4) cellular hyperplasia and then cell death occurs at the interface; and (5) sloughing of dead tissue leaves areas of vacant skeleton between the two grafted individuals. Allograft rejection reactions in sponges have many of the hallmarks of histoincompatibility reactions in higher animals. Although there could be different alloimmune mechanisms operating in different animals, the salient events of allorecognition and antagonistic rejection appear strikingly similar among diverse multicellular animals.     

Investigations of carotenoids in some fauna of the Adriatic Sea. II. Anemonia sulcata and Actinia equina (Coelenterata: Anthozoa)

Investigations have been carried out on the carotenoids in Anemonia sulcata (Pennant) and Actinia equina (L.) (Coelenterata: Anthozoa) from the Adriatic Sea. The presence of the carotenoids was determined by means of columnar and thin-layer chromatography. The following carotenoids were found: Anemonia sulcata: β-carotene, lutein-like (epoxy and free) and astaxanthin; Actinia equina: α-carotene, β-carotene, actinioerythrin, lutein-like (?) and astaxanthin (ester and free).     

Ecology of the northern Red Sea crinoids and their Epi- and Endozoic fauna

Fourteen crinoid species are found along the coral reefs of the northern Red Sea that form aggregations of different species at various depths. The shallowest aggregation consists partly of Lamprometra klunzingeri and Capillaster multiradiatus, occurring subtidal to a depth of 2 to 5 m. Of this group, Heterometra savignii inhabits depths down to 12–15 m. Further down, another group of 10-armed crinoids occurs, dominated by Decametra chadwicki and Oligometra serripinna. The deepest aggregation of crinoids occurs at 45 m; its most common members are Colobometra arabica and Comaster distinctus. Feeding on micro- and nanoplankton, shallow-water crinoids show a circadian activity rhythm; in deeper aggregations (10 to 12 m), this behaviour changes with decreasing illumination, to a diurnal activity rhythm. The symbiotic animals living on crinoids comprise 27 taxons, among them Copepoda (6 species), Mollusca (2 species), Polychaeta (11 species, especially Myzostomida); Ophiurida (1 species), Crustacea, Decapoda (6 species); and the clingfish Lepadichthys lineatus. These symbiotic animals form food webs at various depths, according to the distribution of their crinoid host. The occurrence of Indo-West Pacific and Mediterranean commensals on the same crinoid in the Gulf of Aqaba is of special interest.     

Effects of gamma-aminobutyric acid on the settlement of larvae of the black chiton Katharina tunicata

Laboratory-cultured larvae of the black chiton Katharina tunicata (Wood), collected from central California in May and June of 1979 and 1980, settled preferentially on the encrusting coralline alga Lithothamnium sp. Metamorphosis, which involves the loss of the prototrochal ciliary band, occurred within a period of 2.5 h after larvae began crawling upon the alga. In response to gamma-aminobutyric acid (GABA) at a concentration of 10^-6 M, the lecithotrophic trochophore larvae ceased swimming and settled rapidly. Developmental metamorphosis did not follow settlement in the absence of encrusting coralline alga. However, when both GABA and Lithothamnium sp. were present, larval metamorphosis occurred within 2 h of settlement. Significantly different rates of settlement exist for sibling larvae of different ages; 12 d-old larvae responded more rapily and in greater numbers to treatment with GABA than 10 d-old larvae. Differences in settling rates are attributed to the onset of metamorphic competence.     

Metabolic adaptation of the intertidal worm Sipunculus nudus to functional and environmental hypoxia

The scope of anaerobic metabolism of Sipunculus nudus L. was assessed from the maximal activities of some enzymes of the intermediary metabolism and from the concentration of some metabolites accumulated during enhanced muscular activity and during prolonged experimental hypoxia.

1. Maximal enzyme activities demonstrate that the scope of anaerobic glycolysis, as indicated by maximal activities of glycogen phosphorylase (0.84 U g^-1 fresh wt), far exceeds the aerobic capacity, which is assumed not to surpass the activity of succinate dehydrogenase (0.09 U g^-1 fresh wt). Three pyruvate reductase activities (alanopine-, strombine- and octopine dehydrogenase) can possibly terminate anaerobic glycolysis.
2. During muscular activity, energy is provided by the degradation of phospho-L-arginine and by anaerobic glycolysis. Octopine is the major endproduct during functional anaerobiosis while the formation of strombine is less pronounced.
3. During exposure to a nitrogen atmosphere, several anaerobic endproducts are found to accumulate. Anaerobic glycolysis is terminated by strombine synthesis. This opine accumulates in concentrations much higher than octopine. In addition the concentrations of succinate, propionate and acetate are found to increase in tissues, and/or in the coelomic fluid and the incubation water.
4. The relative contribution of energy by the different anaerobic metabolic pathways are estimated during functional and environmental hypoxia.

     

New epizooic symbioses between sponges of the genera Plakortis and Xestospongia in cryptic habitats of the Caribbean

Three new cases of sponge symbiosis between species of Plakortis and Xestospongia were found in reef caves and mesophotic reef habitats of the Caribbean. Plakortis sp. 1 from the Bahamas associates exclusively with Xestospongia deweerdtae which was originally described living freely on the deep fore-reef and caves of Jamaica. In addition, we found Plakortis sp. 2 from Puerto Rico which associates with both X. deweerdtae and a different Xestospongia sp. Sponge specimens were identified using cytochrome oxidase subunit 1, 28S rRNA and 18S rRNA gene sequence fragments, spicule analysis, and histological sections with SEM. Unlike previous sponge pairs, Xestospongia spp. not only grew as a thin veneer of tissue over the Plakortis host sponge but through the mesohyl, forming inner channels (0.1–1 cm) that may provide a benefit by facilitating more efficient water transport through the dense Plakortis tissue. Symbioses with both Plakortis spp. were documented from an early recruit stage through adulthood. Spicule measurements conducted on symbiotic versus free-living X. deweerdtae revealed significantly smaller spicule sizes for symbiotic individuals, suggesting a cost in terms of silicon availability, or a benefit in terms of a lower investment in skeleton synthesis for support. This study reveals new specialized symbiotic associations between distantly related sponge genera that likely represent an alternative strategy of adaptation for life in reef caves and mesophotic reefs.     

Estimation of coral growth-rates from laboratory 45Ca-incorporation rates

Laboratory ⁴⁵Ca-incorporation rates in hermatypic coral skeletons have previously been used successfully as an index of physiological function. This laboratory method would become more meanigful if it also provided an absolute measure of coral growth rates. In two coral species, Porites compressa and Pocillopora damicornis, ⁴⁵Ca incorporation rates were obtained from short (0.5 h) laboratory incubations using apical (determined as fast growing) portions of freshly collected coral branches. ⁴⁵Ca exchange across the coenosarc was not significant and not corrected for, whereas diurnal fluctuation in ⁴⁵Ca in Pocillopora damicornis was significant and a necessary correction. A calculated surface area is used to express calcification rate. Typical growth rates calculated from the ⁴⁵Ca-incorporation rates were 20 and 6 mm/year for Porites compressa and Pocillopora damicornis, respectively. These rates are considerably higher than those previously obtained in the laboratory, and compare favorably with field growth rates — 24 and 14 mm/year, respectively.     

Decarboxylation of environmental L-leucine by marine red algae

The uptake and metabolism of ¹⁴C-L-leucine by Cystoclonium purpureum (Huds) Batt. (Rhodophyllidaceae) at concentrations of 0.5–50 μM were studied. Leucine was taken up from seawater at rates of 10–160 nmol·g^-1 fresh weight·h^-1. At low leucine levels ( 5 μM) the amino acid was preferably incorporated into cellular protein; amine formation was less than 5%. However at leucine concentrations of 50 μM, more than 30% of the total leucine taken up was decarboxylated to 3-methylbutylamine. This effect is expressed even more in Dumontia increassata (O. F. Müll) Lamour. (Dumontiaceae). In both species some of the amine occurred as a non-volatile derivative (bound amine) from which free amine could quantitatively be released by acid hydrolysis. Leucine and amine uptake were inhibited by CCCP. However, from cells preloaded with both leucine and amine, only free amine is released in the presence of CCCP. The results indicate that decarboxylase activity will be switched on at increased levels of amino acids in seawater which may be expected frequently in habitats of littoral algae (tidal zone). A possible function of amine formation and release within the scope of chemical defence of benthic algae is discussed.     

Reproduction and development of Sabella media,a sabellid polychaete with extratubular brooding

Known reproductive patterns in the polychaete family Sabellidae include: (1) broadcasting of gametes, (2) depositing of benthic egg masses, (3) brooding outside the lip of the tube, and (4) brooding within the tube. There is little information for sabellids in the third category; there-fore we have studied the reproductive biology of Sabella (=Demonax) media (Bush), one of the few species known to brood its early developmental stages outside the tube. A total of 32 adults of S. media with egg cocoons were collected from the surfaces of compound ascidians (Aplidium sp. and Didemnum sp.) and from holdfasts of a brown alga (Hedophyllum sessile) at Eagle Cove, San Juan Island, Washington, USA, from June 1981 through June 1982, between tidal levels of 0.0 and 0.6 m. The donutshaped cocoons, each containing over 1 000 eggs, are deposited just outside the tube aperture from April through September. Larval development takes place within the cocoon through the 3-setiger stage, at which time the larvae emerge (about 8 d after egg deposition). The newly hatched larvae are demersal and swim near the bottom of the culture dish for 1 d before settlement. Metamorphosis takes nearly a week, and initial formation of the tube is also gradual. The reproductive patterns within the family Sabellidae are discussed, and the adaptive significance of extratubular brooding considered.     

Assimilation,inter-organ transfer and excretion of americium in two teleost fish

Radiotracer experiments were performed (February–April, 1982) to study the assimilation and metabolism of the transuranium nuclide americium-241 in the marine teleosts Serranus scriba (Linnaeus, 1758) and Scorpaena notata Rafinesque, 1810, caught off the Monaco coast. Fish fed with ²⁴¹Am-labelled food showed that assimilation of this radionuclide takes place through the gastrointestinal walls and that the small fraction accumulated is incorporated mainly in the skin, muscle and skeleton. Gut-transfer coefficients were similar in both species and averaged 0.7% (range 0.1 to 1.7%) of the ingested activity. The calculated biological half-lives for loss of the absorbed fraction ranged between 49 and 61 d for Serranus scriba and 12 and 117 d for Scorpaena notata. Results from an intramuscular injection experiment indicated that ²⁴¹Am was retained mainly in the liver, skin and skeleton; the fraction accumulated by muscle was very low. Liver displayed a relatively short biological half-time for ²⁴¹Am loss of roughly 24 d. Routes of ²⁴¹Am excretion from the teleosts appear to be through the kidneys, gills and feces with bile serving as a possible excretion route from the liver. From the limited amount of published information available for comparison, experimental evidence is presented which suggests that ²⁴¹Am taken up via the food chain is more biologically available to marine fish than is plutonium.     

Habitat selection in the tropical polychaeteSpirobranchus giganteus

Total body weight, tube length, abdomen weight and branchial crown weight of individualSpirobranchus giganteus (Pallas) living on four different coral species on the bank reef on the west coast of Barbados were investigated in 1986. Worms onDiploria strigosa were larger in all size parameters than those onMontastrea annularis, which were larger than those onMontastrea cavernosa, which were larger than those onPorites porites. The differences between worms onM. annularis andM. cavernosa were not significant. Variation in worm size on the different corals could result from variation in mortality and/or variation in growth. Whichever the case, larger worms may have higher lifetime reproductive success. Abdomen weight is an index of gamete production inS. giganteus, and increases with increasing body weight for worms on all corals. Moreover, larger worms do not have lower abdomen weight at a given body size than smaller worms. This may imply that faster growth does not reduce gamete production at body size. The coral species on which worms are largest are those most preferred by worm larvae in the laboratory and most heavily colonised by adult worms in the field. The results suggest adaptive habitat selection by planktonic larvae ofS. giganteus.     

A light and electron microscope investigation of gametogenesis in Nicolea zostericola (Polychaeta: Terebellidae)

A light and electron microscope study was conducted on Nicolea zostericola (Oersted: Grube, 1860) to describe the processes of gametogenesis. N. zostericola is a monotelic species which spawns from 2 to 5 times over a period of 2 weeks at the end of its 1-year life span. The gonads consist of plates of cells bulging from beneath the peritoneum in the thoracic region of the body. Oogenesis has been divided into 3 phases: an ovarian phase in which gametogonia and primary oocytes are retained within the ovary, a follicular phase, in which oocytes are ovulated into the coelom surrounded by several layers of follicle cells and the vitellogenic phase, where individual oocytes rupture through the follicle cells and enter the coelomic fluid where cell growth and yolk synthesis occur. The combined ovarian and follicular phases have a duration of about 3 to 5 weeks and the vitellogenic phase about 5 to 6 months. Vitellogenesis is described on the ultrastructural level. Yolk synthesis involves the participation of the Golgi apparatus and endoplasmic reticulum of the oocyte in possible combination with nutrient absorption from the coelomic fluid. Spermatogenesis involves the rupturing of masses of spermatogonia into the coelom from the testis, surrounded by peritoneal cells. Smaller clusters of spermatocytes are later released into the coelomic fluid where mitotic and meiotic divisions eventually produce flattened plates of sperm. The mature spermatozoan possesses an elongate head, atypical for most polychaetes.     

Shell parameters in relation to zonation in mediterranean limpets

The relation of shell form to habitat in the Mediterranean limpets Patella caerulea (L.) and P. lusitanica (Gmelin) was investigated. P. caerulea shells were significantly longer, broader, but lower in height than P. lusitanica shells. The regression of the ratio (length plus breadth)/2 on the height of the shell was found to characterize the two limpet populations, with a small degree of overlap reflecting the distribution of the two limpets on the shore.     

Assimilation of symbiont-derived photosynthates in some solitary and colonial radiolaria

Evidence for host assimilation of ¹⁴C-labeled symbiont photosynthates is presented from laboratory studies of the solitary radiolarian Thalassicolla nucleata and the colonial species Collosphaera huxleyi. The amount of ¹⁴C-labeled product assimilated in the central capsule of T. nucleata is directly related to the amount of ¹⁴C incorporated by the symbionts. In C. huxleyi central capsules, the percentage of ¹⁴C-label occurring in the water-soluble fraction is 38% and in the lipid-soluble fraction is 20%, the remainder being in insoluble products. Within the lipid-soluble fraction, a substantial percentage of the ¹⁴C activity is associated with the triglyceride and wax ester fractions. The significance of these findings is discussed in relation to the possible physiological role of symbionts in sustaining the host and stabilizing the host-symbiont association.