Fungi in corals: black bands and density-banding of Porites lutea and P. lobata skeleton

 Dark coloration of coral skeleton forming black bands is commonly observed in fractured, massive-coral colonies (Porites lutea and P. lobata) collected from May- otte Island in the Mozambique Channel and Moorea Island in French Polynesia. Black-banding was similar in corals from the two areas and was associated with an assemblage of microbial endoliths: Ostreobium queketti, a common siphonal chlorophyte, and a type of Aspergillus-like fungus. Fungi of coral skeletons are capable of euendolithic growth entirely within the skeleton, and of cryptoendolithic growth whereby they spread from the skeleton into the skeletal pores. The morphology and size of fungal hyphae differs significantly between euendolithic, cryptoendolithic and reproductive phases. Reproductive phases involve formation of conidiophores. Insoluble residues in black bands involve a dark pigment and a dark membranous veil. When attacked by fungi, the algae are usually destroyed. They darken and are threaded by dense, dark-brown, fibrous excrescences. The fungi excrete a dark pigment that stains the surrounding skeletal carbonate black. The pigment is organic, and its presence correlates with higher concentrations of polysaccharides. Black bands match high-density bands of the coral skeleton. Both black bands and high-density bands form at the end of the rainy season in Mayotte. Thus, black-banding in the corals studied is caused by a series of events, beginning with an increase in the abundance of endolithic algae followed by an increase in skeletal density. The algae are then attacked by fungi, which produce more cryptoendolithic hyphae and conidia that are associated with production of the dark pigment. Received: 29 January 1999 / Accepted: 29 September 1999     

Exploring the role of chitinolytic enzymes in the sea fan coral, <Emphasis Type="Italic">Gorgonia ventalina</Emphasis>

Chitinases are involved in defense against chitinaceous pathogens in both invertebrates and vertebrates. This study investigated whether sea fan corals, Gorgonia ventalina (Linnaeus) collected from the Florida Keys between the summer of 2002 and the summer of 2005 contain chitinases, and whether these enzymes could serve an analogous protective role against the fungal pathogen, Aspergillus sydowii (Thom et Church). Crude extracts of healthy sea fans contained detectible levels of exochitinase activity in an in vitro microplate assay using fluorogenic substrates. The exochitinase levels decreased upon injury, agitation, or manipulation of the tissue. A concurrent, transient increase of exochitinase in the surrounding water suggests that sea fans release chitinases as a response to these stresses. By contrast, endochitinase was detected in only 2 of 15 sea fans (13%), suggesting a high degree of variation for this enzyme. Sea fan chitinase-containing seawater and anion exchange chromatography fractions were both active against A. sydowii in an absorbance-based antifungal assay. The presence of chitinases in sea fan extracts, their release into the surrounding water upon stress, and their activity against A. sydowii suggests that further study of these enzymes in coral stress responses is warranted.     

Determination of canthaxanthin in the red coral (<Emphasis Type="Italic">Corallium rubrum</Emphasis>) from Marseille by HPLC combined with UV and MS detection

Corallium rubrum is the most famous and precious coral due to the intense characteristic red colour of its skeleton. We have determined the presence of carotenoids in natural samples of C. rubrum from Marseille, Riou, and investigated their chemical composition. Analysis was performed both on soft tissues and hard tissues including spicules and skeleton. Since hard tissues are made of a mineral fraction and an organic fraction obtained after demineralization, extraction was performed both with and without demineralization by EDTA. The extracts were analyzed by complementary methods of TLC, HPLC/DAD and HPLC/MS. The components were separated by RP-18 chromatography column using acetonitrile/methanol for HPLC/DAD and acetonitrile/water gradient for HPLC/MS analysis. Our results give the first evidence of the presence of canthaxanthin, 4,4′-diketo-β-carotene, as the major carotenoid in all samples. Spicules exhibited higher content in carotenoids than skeleton. Demineralization treatment improved the efficiency of carotenoid extraction by an average factor of 5 and showed that organic matrix contains canthaxanthin.     

Monthly skeletal extension rates of the hermatypic corals<Emphasis Type="Italic"> Montastraea annularis</Emphasis> and<Emphasis Type="Italic"> Montastraea faveolata</Emphasis>: biological and environmental controls

Monthly skeletal extension rates were measured in colonies of Montastraea annularis and M. faveolata growing at Mahahual and Chinchorro Bank, in the Mexican Caribbean. Temperature, light extinction coefficient (k_d), sedimentation rate, dissolved nutrients and wave energy were used as indicators of environmental conditions for coral growth. Zooxanthella density and mitotic index, nitrogen, phosphorous and protein in coral tissue, and living tissue thickness were measured during periods of high-density-band (HDB) and low-density-band (LDB) formation. To test their value as indirect measures of competition between zooxanthellae and host, as well as coral health and performance in both species, these biological parameters were also measured, during the HDB-formation period, in corals collected at La Blanquilla. This reef is located in the Gulf of Mexico, in an area of suboptimal environmental conditions for coral growth. M. faveolata had a significantly higher skeletal extension rate than M. annularis. Corals growing in Mahahual had significantly higher skeletal extension rate than those living in Chinchorro Bank. This is consistent with inshore–offshore gradients in growth rates observed by other authors in the same and other coral species. This is probably due to less favorable environmental conditions for coral growth in near shore Mahahual, where there is high hydraulic energy and high sedimentation rate. Contrary to observations of other authors, skeletal extension rate did not differ significantly between HDB- and LDB-formation periods for both species of Montastraea. Both species produced their HDB between July and September, when the seawater temperatures are seasonally higher in the Mexican Caribbean. Tissue thickness indicated that environmental conditions are more favorable for coral health and performance during the HDB-formation period. Mitotic index data support the idea that zooxanthellae have competitive advantages for carbon over the host during the LDB-formation period. So, corals, during the LDB-formation period, with less favorable environmental conditions for coral performance and at a disadvantage for carbon with zooxanthellae, add new skeleton with little or no opportunity for thickening the existing one. This results in an equally extended skeleton with lower density, and the stretching response of skeletal growth, proposed for M. annularis growing under harsher environmental conditions, also occurs during the LDB-formation period.Communicated by P.W. Sammarco, Chauvin     

Lipogenesis in the intact coral Pocillopora capitata and its isolated zooxanthellae: Evidence for a light-driven carbon cycle between symbiont and host

Surface tissue of the reef coral Pocillopora capitata contained approximately 34% lipid on a dry weight basis. Of this, 75% was storage lipid (wax ester and triglyceride) and 25% structural (phospholipid, galactolipid, etc.). Based on chlorophyll a: lipid ratios of intact coral and isolated zooxanthellae, it was determined that over 90% of the storage lipid resided in the host tissue. One half of the structural lipids was found in the host and the other in the symbiotic algae. Gentle fractionation of coral tissue indicated that zooxanthellae possessed less than 14% of the total coral protein. Coral tips and isolated zooxanthellae were incubated with sodium acetate-1-¹⁴C in light and dark to obtain lipogenic rates and proportions of fatty acids and lipid classes synthesized. The rate of lipid synthesis from acetate-1-¹⁴C by intact coral was stimulated three-fold in the light (1200 lux), which indicated that the majority of coral lipogenesis occurred in the zooxanthellae. Intact coral triglycerides contained ca. 68% of the ¹⁴C-activity and wax esters ca. 21%. Zooxanthellae isolated by the Water Pik technique synthesized negligible amounts of wax ester, which implied that wax ester synthesis was a property of the animal tissue. Isolated zooxanthellae and intact coral synthesized identical triglyceride fatty acids from acetate-1-¹⁴C. This study provides evidence for a carbon cycle between host and symbiont whereby the zooxanthellae take up host-derived carbon (probably in the form of acetate), synthesize fatty acids using their photosynthetically derived energy, and return the lipid to the host where it appears as wax ester and triglyceride.     

A novel Vibrio sp. pathogen of the coral Pocillopora damicornis

A coral pathogen was isolated from the diseased tissue of Pocillopora damicornis in Zanzibar. The pathogenic bacterium, referred to as Vibrio coralyticus YB, was classified as a member of the genus Vibrio. Based on its 16S rDNA sequence, V. coralyticus is probably a new species. In controlled aquaria experiments at 26-29°C, inoculation of pure cultures of V. coralyticus YB either into the seawater or by direct contact onto the coral caused tissue lysis of P. damicornis fragments. At 29°C, lysis began as small white spots after 3-5 days, rapidly spreading so that by 2 weeks the entire tissue was destroyed, leaving only the intact bare skeleton. When an infected diseased coral was placed in direct contact with a healthy one, the healthy coral lysed in 2-4 days, further indicating that the disease was contagious. Inoculation with as few as 30 bacteria ml^-1 was sufficient to infect and lyse corals. Seawater temperature was a critical variable for the infectious process: infection and lysis occurred rapidly at 27-29°C, slowly at 26°C and was not observed at 25°C. The data suggest that the presence of V. coralyticus YB, even in low numbers, in seawater surrounding a coral reef will lead to tissue destruction of P. damicornis, when seawater temperatures rise.     

Antimicrobial activity of Red Sea corals

Scleractinian corals and alcyonacean soft corals are the two most dominant groups of benthic marine organisms inhabiting the coral reefs of the Gulf of Eilat, northern Red Sea. Antimicrobial assays performed with extracts of six dominant Red Sea stony corals and six dominant soft corals against marine bacteria isolated from the seawater surrounding the corals revealed considerable variability in antimicrobial activity. The results demonstrated that, while the majority (83%) of Red Sea alcyonacean soft corals exhibited appreciable antimicrobial activity against marine bacteria isolated from the seawater surrounding the corals, the stony corals had little or no antimicrobial activity. From the active soft coral species examined, Xenia macrospiculata exhibited the highest and most potent antimicrobial activity. Bioassay-directed fractionation indicated that the antimicrobial activity was due to the presence of a range of compounds of different polarities. One of these antibiotic compounds was isolated and identified as desoxyhavannahine, with a minimum inhibitory concentration (MIC) of 48 μg ml⁻¹ against a marine bacterium. The results of the current study suggest that soft and hard corals have developed different means to combat potential microbial infections. Alcyonacean soft corals use chemical defense through the production of antibiotic compounds to combat microbial attack, whereas stony corals seem to rely on other means.     

Photobiology of endolithic microorganisms in living coral skeletons: 1. Pigmentation, spectral reflectance and variable chlorophyll fluorescence analysis of endoliths in the massive corals Cyphastrea serailia, Porites lutea and Goniastrea australensis

We used microscopy, reflectance spectroscopy, pigment analysis, and photosynthesis-irradiance curves measured with variable fluorescence techniques to characterise the endolithic communities of phototrophic microorganisms in the skeleton of three massive corals from a shallow reef flat. Microscopic observations and reflectance spectra showed the presence of up to four distinct bands of photosynthetic microorganisms at different depths within the coral skeleton. Endolithic communities closer to the coral surface exhibited higher photosynthetic electron transport rates and a green zone dominated by Ostreobium quekettii nearest the surface had the greatest chlorophyll pigment concentration. However, Ostreobium was also present and photosynthetically active in the colourless band between the coral tissue and the green band. The spectral properties and pigment density of the endolithic bands were also found to closely correlate to photosynthetic rates as assessed by fluorometry. All endolithic communities were extremely shade-adapted, and photosynthesis was saturated at irradiances <7 μmol photons m⁻²s⁻¹.     

Association of <Emphasis Type="Italic">Waminoa</Emphasis> sp. (Acoela) with corals in the Wakatobi Marine Park,South-East Sulawesi,Indonesia

This is the first quantitative study on the prevalence of epizoic Waminoa sp. acoel worms and their association with corals in the Wakatobi Marine National Park (WMNP), South-East Sulawesi, Indonesia. Three replicate transects were laid on the reef crest, flat and slope at six sites in 2006 and eight sites in 2007. Four of the sites were common in both years. In total 69 transects were surveyed in 2006, and 87 transects in 2007. A total of 4.8% of all observed hard corals were associated with acoel worms in 2006 and 2.6% of hard and soft corals in 2007. Acoels were present on 16 and 21 of the coral taxa studied in 2006 and 2007 respectively. The worms were strongly associated with the azooxanthellate coral Tubastrea spp. and were rare or absent on the most abundant coral genera Montipora and Porites. The mean number of corals having acoels was highest on reef slopes, whereas acoels were virtually absent on reef flats. Corals that had a high and a medium cover of worms were more common in 2007 than in 2006. No significant trend in the adaptation of the zooxanthellae of Waminoa sp. to different depths at different sites was revealed. The impact of the worm on the coral is unknown, but high numbers may have a shading effect and a negative impact on the coral’s photophysiology. This acoel merits more study of its life cycle, its photophysiology, and its impact on its host corals. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Frequency of sublethal injury in a deepwater ophiuroid, <Emphasis Type="Italic">Ophiacantha bidentata</Emphasis>, an important component of western Atlantic <Emphasis Type="Italic">Lophelia</Emphasis> reef communities

The occurrence and relative abundance of tissue (arm) regeneration in the ophiuroid, Ophiacantha bidentata (Retzius), was examined in individuals collected primarily among colonies of the deep-water coral Lophelia pertusa off the southeastern United States. Seven deep-water coral sites (384–756 m), located between Cape Lookout, NC, and Cape Canaveral, FL, were sampled in June 2004 using a manned submersible. The presence of regenerative tissue was evaluated by visual inspection of each individual ophiuroid, and the proportion of regenerating arms per individual was examined relative to size of individual, geographic location, and depth of collection. Ophiacantha bidentata, the dominant brittle star collected, commonly displayed signs of sublethal injury with over 60% of individuals displaying some evidence of regeneration. These levels of regeneration rival those reported for shallow-water ophiuroids. Larger individuals (>6.5 mm disc size) had a higher incidence of regeneration than smaller individuals. Size of individual and percent of regeneration were negatively correlated with depth. Although O. bidentata was significantly less abundant in southern versus northern sites, ophiuroid abundance did not appear to be influenced by amount or density of coral substratum. Presence of dense aggregations of O. bidentata indicates that they are an important component of the invertebrate assemblage associated with deep-water coral habitat especially in the northern part of the study area. Assuming that observed frequencies of injury and subsequent regeneration represent predation events then dense ophiuroid aggregations in deep-water coral habitats represent an important renewable trophic resource within these communities.     

Studies on reef corals. III. Fine structural changes of calicoblast cells in Pocillopora damicornis during settling and calcification

The site of reef-coral calcification has been studied in the branching coral Pocillopora damicornis Lamarck. Electron microscopy and X-ray microprobe analysis were performed on the calicoblast epidermis of newly settled larval stages and of adult coral. During settling, the heterogeneous columnar cell composition of the planktonic larva epidermis is replaced by a simple epithelium consisting of a single cell type, the calicoblast cell. Metamorphosis appears tightly linked to settling, with cell changes occurring within hours after attachment, and is marked by the appearance of a new secretory cell. The calicoblast cell of the adult coral is extremely flattened, and interdigitates extensively with adjacent calicoblast cells. This cell possesses a featureless plasma membrane lacking microvilli or flagella. It characteristically contains large membrane-bound vesicles with homogeneously fine granular contents. Preliminary microprobe analysis indicated a higher calcium content in these vesicles than in surrounding tissue; however, not in concentrations suggesting calcium-carbonate precipitation. They may represent sites of organic matrix synthesis. The calicoblast epidermis is separated from the underlying coral skeleton by a narrow gap. This gap appeared devoid of substructure, either organic or inorganic. The coral soft tissues are attached to the skeleton by mesogleal attachment processes, the desmoidal processes. These consist of a complex fibrous network originating in the mesoglea, and inserting onto the skeleton via specialized attachment regions consisting of electron-opaque membranous plaques. Skeletogenesis in reef-corals probably occurs extra-cellularly, external to the calicoblast epidermis, by simple overgrowth of the skeleton.     

Fungal disease resistance of Caribbean sea fan corals (Gorgonia spp.)

An on-going, Caribbean-wide epizootic affecting sea fan corals (Gorgonia spp.) is caused by the fungus Aspergillus sydowii (Thom et Church). We examined the role of crude extracts in resistance of two species of sea fans, Gorgonia ventalina (L.) and G. flabellum (L.), against A. sydowii and a bacterial pathogen of fish, Listonella anguillarum (MacDonell et Colwell). Sea fans were collected in January 1997 from San Salvador, Bahamas, and in June 1997 and January 1998 from Alligator Reef, Florida Keys, USA. Crude extracts from both species were tested to determine concentrations inhibiting germination of A. sydowii spores. Crude extracts from both species inhibited spore germination at concentrations as low as 1.5 mg ml⁻¹; most samples were active at 5 to 10 mg ml⁻¹. These concentrations are within the range estimated in living tissue and were higher in healthy colonies suggesting their role in mediating disease susceptibility. We also detected within-colony gradients in antifungal activity, which varied with the disease state of the colony. In healthy sea fans, resistance was highest at colony edges and lowest in medial and central regions of the colony. Among sea fans with lesions in the colony center, resistance in tissue from proximal and medial regions was as high as tissue from the colony edge (i.e. distal region). The increase in antifungal activity suggests an inducible response by the coral host to the fungal pathogen. This response is most evident among sea fans with lesions in the colony center and not among colonies with lesions at the edge. Antibacterial activity of crude extracts against L. anguillarum was highest at the colony edge but did not vary with disease state or tissue location. Received: 10 March 1999 / Accepted: 10 November 1999     

Short-term growth measurements of corals using an accurate buoyant weighing technique

An accurate method for determining the growth rates of the skeleton of isolated branch tips (nubbins) of corals over intervals of less than 24 h is described. The skeletal weight of the coral was estimated from its buoyant weight in seawater whose density had been accurately determined. The coral tissues accounted for between 1 and 5% of the total buoyant weight in Pocillopora verrucosa and Acropora humilis with differing relative tissue biomass. After correcting for tissue buoyant weight, predictions of skeletal weight were accurate to within 1%. The method was used to estimate the growth of sample nubbins of Porites porites of similar diameter, in 2 m of water at Discovery Bay, Jamaica. Since growth of these branch tips is apical, growth rate could be expressed without correction for the size. The mean 24 h skeletal growth rate ranged between 40 and 47 mg. Differences could be measured between day-time and night-time growth, the day: night ratio being 3.7. The method also showed that P. porites virtually ceases calcification during the 4 to 5 d periods that it becomes enclosed in a mucus tunic. Nubbins of P. porites attached to the reef at different locations showed clear differences in growth rate with depth, and between clear and turbid water sites. The growth rate of nubbins was compared with that of branch tips of whole corals by measuring the linear extension after staining with Alizarin Red S. After 3 1/2 mo, the mean linear extension was 4.1 mm in each case, indicating that the growth rate of nubbins is the same as that of branch tips of the whole colony. It is suggested that this buoyant weighing technique will find applications in laboratory experiments with calcification mechanisms and as a bioassay on reefs exposed to environmental stress.Contribution No. 464 of the Discovery Bay Marine Laboratory     

Effects of diterpenes derived from the soft coral Sinularia flexibilis on the eggs,sperm and embryos of the scleractinian corals Montipora digitata and Acropora tenuis

Sinularia flexibilis Quoy and Gaimard, 1833 is a dominant soft coral on many Indo-Pacific coral reefs, and has been found to release toxic compounds (diterpenes), which cause tissue necrosis and death in nearby scleractinian corals. This study investigates how S. flexibilis-derived diterpenes inhibit the development of Acropora tenuis Dana, 1846 and Montipora digitata Dana, 1846 eggs and larvae in vitro. Collection and experimental sites at Magnetic (146°49'E; 19°8'S) and Orpheus (146°28'E; 18°32'S) Islands, Queensland, Australia, were utilized during the spawning seasons of 1989–1992. Freshly spawned coral eggs were placed in solutions of three different terpenes, flexibilide, dihydroflexibilide and sinulariolide, at 5 and 10 ppm, before, during, and after fertilisation. The majority of eggs which were fertilised in the presence of the diterpenes lost their cellular integrity and burst just a few hours after treatment. Terpenes were not toxic to unfertilised eggs, nor to 24 h-old embryos, although sperm ceased swimming activity after 1 h of treatment. The terpenes were not fatal to the sperm because fertilisation still occurred in their presence. The ability of the soft coral-derived diterpenes to inhibit cell division suggests that they may have potential applications in cancer chemotherapy.     

Population structure and feeding deterrence in three shallow-water antarctic soft corals

Alcyonium paessleri and Clavularia frankliniana are numerically abundant soft corals in the nearshore (12 to 33 m depth) benthic communities of eastern McMurdo Sound. They are much less abundant in western McMurdo Sound where a third species, Gersemia antarctica, co-occurs in low numbers. The body tissues of these three species are comprised mainly of organic material (53 to 70% dry wt), which is primarily dervied from NaOH-soluble protein and refractory material. The energetic contents of the whole-body tissues of A. paessleri, C. frankliniana and G. antarctica are 15.9, 17.3, and 14.5 kJ g^-1 dry wt, respectively. The mean biomass per individual is 1.81, 0.008, and 45 g dry wt for each respective species. Based on population densities of 7.3, 1337.3, and 0.04 soft corals m^-2 for A. paessleri, C. frankliniana and G. antarctica, respectively, the population energetic densities are estimated to be 210.1, 185.1, and 26.1 kJ m^-2. Despite the relatively rich energetic content of the tissue and apparent vulnerability to predators, very little predation occurs on these soft corals. Two potential predators, the antarctic sea stars Perknaster fuscus and Odontaster validus, exhibited significant chemotactic defensive tube-foot retractions to hexane, chloroform, methanol, and aqueous methanol extracts of each soft coral. In addition, wholebody tissue of each soft coral was rejected by the demersal fish Pseudotrematomus bernacchii and the cryopelagic fish Pagothenia borchgrevinki. In contrast, whole soft-coral tissues sequentially extracted in four increasingly polar solvents were readily ingested by these antarctic fishes, indicating that sclerites do not play a significant role in deterring predators. Our results indicate that these antarctic soft corals contain bioactive compounds which deter common predatory seastars and fishes.     

Porites and the Phoenix effect: unprecedented recovery after a mass coral bleaching event at Rangiroa Atoll,French Polynesia

The 1997/1998 El Niño Southern Oscillation (ENSO) was the most severe coral bleaching event in recent history, resulting in the loss of 16 % of the world’s coral reefs. Mortality was particularly severe in French Polynesia, where unprecedented mortality of massive Porites was observed in lagoonal sites of Rangiroa Atoll. To assess the recovery of massive Porites 15 years later, we resurveyed the size structure and extent of partial mortality of massive Porites at Tivaru (Rangiroa). Surveys revealed an abundance of massive Porites colonies rising from the shallow lagoonal floor. Colony width averaged 2.65 m, reaching a maximum of 7.1 m (estimated age of ~391 ± 107 years old). The relative cover of recently dead skeleton within quadrats declined from 42.8 % in 1998 to zero in 2013, yet the relative cover of old dead skeleton increased only marginally from 22.1 % in 1998 to 26.1 % in 2013. At a colony level, the proportion of Porites dominated by living tissue increased from 34.9 % in 1998 to 73.9 % in 2013, indicating rapid recovery of recent dead skeleton to living tissue rather than transitioning to old dead skeleton. Such rapid post-bleaching recovery is unprecedented in massive Porites and resulted from remarkable self-regeneration termed the ‘Phoenix effect’, whereby remnant cryptic patches of tissue that survived the 1997/1998 ENSO event regenerated and rapidly overgrew adjacent dead skeleton. Contrary to our earlier predictions, not only are large massive Porites relatively resistant to stress, they appear to have a remarkable capacity for recovery even after severe partial mortality.     

Dinoflagellate <Emphasis Type="Italic">Alexandrium leei</Emphasis> (Dinophyceae) from Singapore coastal waters produces a water-soluble ichthyotoxin

A strain of Alexandrium leei Balech that was isolated in October 2002 from Singapore coastal waters and identified by light and scanning electron microscopy and phylogenetic analysis using LSU rDNA sequences, is toxic to Asian sea bass fingerlings (Lates calcarifer Bloch). The ichthyotoxicity of the algal cells obtained by filtration (15 μm mesh net) and rinsed with sterile culture medium indicated that the toxicity of A. leei was probably not due to bacterial contamination, which was further supported by a negative correlation between the time to death of the fish and the dosage of algal cells applied. Paralytic shellfish toxins (PST) could not be detected indicating that PST was not the cause of fish mortality. Fish bioassays using frozen culture, heat-treated cultures, cell-free culture medium, and hexane, ethyl acetate, and water extracts of algal cells indicated that A. leei produces a heat-stable, polar ichthyotoxin(s) which can be released from the algal cells into the culture medium. Phylogenetic analysis based on the LSU rDNA sequences of A. leei confirmed its identification and indicated that the Singapore strain is more similar to isolates from Malaysia than to a geographically distant strain from Korea. This is the first evidence of icthyotoxin production by A. leei.     

Reproduction, larval development and functional relationships of the burrowing, spionid polychaete Dipolydora armata with the calcareous hydrozoan Millepora complanata

Dipolydora armata (Langerhans, 1880) is a small (4 to 5 mm) spionid polychaete found burrowing in the calcareous hydrozoan Millepora complanata Lamarck, 1816, on coral reefs at Barbados, West Indies. It excavates complex networks of interconnecting burrows and forms aggregations of worms in cavities within branches of the coral. Adult worms have a mixed feeding mode (suspension feeding and deposit feeding). Size–frequency distributions of worms in branch samples suggest that they mature in a single year and that reproduction occurs throughout the year. Burrow openings on the surface of the coral develop distinctive, erect spines caused by combined growth of worm tubes and host tissue. Millepore zooids were absent in the vicinity of tube openings and on spines, and thus the potential feeding surface of the coral will be reduced in heavily colonized branches. Burrows and openings were densest at the bases of millepore branches where weakening of the skeleton would be expected to occur. The absence of openings near the branch tips suggests difficulty in larval settlement there, amongst stinging zooids. Reproduction␣and larval development of the worms were examined, and a sequence of larval stages from one to 20 segments and a juvenile stage of 22 segments are described. Eggs are deposited in brood sacs attached to the burrow wall, and the larvae feed upon nurse eggs (adelphophagy). The presence of larvae and juveniles occurring free in the burrows suggests that larval development may be completed within the host coral as an alternative or in addition to a planktonic larval phase. Lack of provisional larval setae, early development of adult capillary setae, production of special spermatophores and a protracted breeding cycle in D. armata are all traits which would favour complete development within the host skeleton. Received: 6 March 1997 / Accepted: 25 October 1997     

Pit structure and trophic relationship of the coral pit crab Cryptochirus coralliodytes

Individuals of the pit crabs Cryptochirus coralliodytes Heller inhabit massive corals of the family Faviidae. Their pit walls were observed to be covered by blue-green algae and fungi. We suggest that the crabs enhance the growth of these algae and fungi with their metabolic excretions, which contain ammonium. The endolithic algae and the fungi may facilitate the abrasion of the coral skeleton by the crabs, by perforating it and thus weakening the skeletal structure. Computerized tomography analysis revealed dense skeletal material around the pits. Transverse sections showed that the calcification around the pit was similar to other parts of the colony, whereas the macro-architecture was different. Such a difference is the result of the crabs' influence on the corals' living tissue, possibly on the calicoblast which deposits the coenosteum. Crabs, which were exposed to carbon-labeled corals for 1, 7 and 18 d, accumulated labeled carbon, indicating transfer of carbon from the coral tissue to the crabs. Histochemical examination of the stomach and gut of crabs revealed the presence of mucopolysaccharids in the gut, supporting the hypothesis that the crabs eat coral products. The findings of this study provide additional evidence that C. coralliodytes are parasites and support the general hypothesis that a nutritional relationship may have served as a basis for selection. Received: 20 October 1998 / Accepted: 29 April 1999