Genetic differentiation between morphotypes of the marine sponge Suberites ficus (Demospongiae: Hadromerida)

Sponges of three morphotypes of Suberites ficus (Johnston, 1842) were collected during February and March 1985 off the south-west of the Isle of Man, and were compared by using spicule size distributions and genetic allele frequencies of isozyme loci. The populations did not show any significant differences of spicule size or type, but could be easily differentiated into three separate species based on isozyme patterns. Samples of pale orange S. ficus growing on gastropod shells inhabited by hermit crabs (Pagurus spp.) were reproductively isolated from the redorange and the pale yellow colour morphs encrusting the bivalve Chlamys opercularis. These latter two colour morphs were genetically similar, but significant differences were observed at two of the 19 gene loci assayed. All the sponges studied were sympatric, and therefore the genetic differences, indicating reproductive isolation, are strong evidence for separate gene pools and, hence, that they are different species. The genetic identity between the two colour morphs of S. ficus on C. opercularis shells was 0.977, whilst between each of these and S. ficus on hermit crabs it was about 0.65. In all three species genetic variability was high, with mean expected and observed heterozygosity values per locus ranging from 0.17 to 0.36.     

The systematic position of some boring sponges (Demospongiae,Hadromerida) studied by molecular analysis

A phylogenetic analysis of some bioeroding sponges of the family Clionaidae (Order Hadromerida) was performed to resolve some taxonomic problems both at the specific and the supraspecific level using the D2 and D3 regions of 28S rDNA. Species belonging to the genera Cliona, Cliothosa, Spheciospongia (fam. Clionaidae) and Diplastrella (fam. Spirastrellidae) from the Mediterranean Sea and Celebes Sea (Indonesia) were analysed. In the phylogenetic tree, the species clustered on two main branches, one comprising Cliona celata, C. rhodensis, C. utricularis, and Cliothosa hancocki, and the other made up of C. viridis, C. nigricans, C. schmidti, C. jullieni, Spheciospongia solida and S. vagabunda. Above the species level, data do not support the separation of the genus Cliothosa from Cliona, while they do support the inclusion of some massive boring species, previously assigned to the genus Spirastrella, in the family Clionaidae. At the species level, data demonstrated the genetic identity of taxa C. viridis and C. nigricans, in spite of their considerable morphological differences. In contrast, the yellow species commonly attributed to C. celata are probably to be considered as a complex of sibling species with a number of distinct taxa present in the Mediterranean. Data also showed the identity of the Mediterranean and Pacific populations of C. schmidti, suggesting the status of a Tethyan relict for this species.     

Unravelling host and symbiont phylogenies of halichondrid sponges (Demospongiae, Porifera) using a mitochondrial marker

We present the first comparative phylogenetic analysis of a selected set of marine sponges and their bacterial associates. The Halichondrida form an important order in demosponge systematics and are of a particular interest due to the production of secondary metabolites. We sequenced a fragment of the cytochrome oxidase subunit 1 (CO1) gene of the sponges and their bacterial associates, compared the reconstructed phylogenies and found evidence for radiation in coevolution. The tree of six host-species associations showed four supported cospeciation events between the sponges and the bacteria. In addition, we present the first gene tree of sponges based on a mitochondrial marker. The tree shows major congruences with previous morphological studies and suggests the applicability of a mitochondrial marker in sponge molecular systematics.     

Gametogenesis,embryogenesis, and larval features of the oviparous sponge <Emphasis Type="Italic">Petrosia ficiformis</Emphasis> (Haplosclerida,Demospongiae)

This study describes the reproductive cycle of Petrosia ficiformis and documents for the first time embryogenesis in an oviparous haplosclerid demosponge. Gonochoric adults, occurring in 2:1 female/male ratios, spawned in late autumn, after a 7-month long oogenesis and a 2.5-week-long spermatogenesis. Following a remarkable migration, the oocytes were released as 250 μm eggs bearing attached polar bodies and a thin mucous cover. Round-headed spermatozoa with three large mitochondria, and many proacrosomal vesicles fertilized the eggs externally. A fertilization membrane appeared around the zygotes. Nearly equal and total cleavage led to a stereoblastula that subsequently became an entirely ciliated larva. The larva alternated spherical and multilobate body shape and consisted of undifferentiated cells. It had poor abilities to swim and glide, probably experiencing passive dispersal and unselective attachment. Settlers developed choanocyte chambers after 1.5 months. Symbiotic microbes were absent from gametes and larvae, being necessarily acquired from the ambient at each sponge generation. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Ultrastructural aspects of the symbiosis between two species of the genus Aphanocapsa (Cyanophyceae) and Ircinia variabilis (Demospongiae)

The association between two species of the genus Aphanocapsa (Cyanophyceae) and the sponge Ircinia variabilis has been studied by electron microscopy. A. feldmanni is localized in the mesohyl or inside the cells of the sponge, while the larger A. raspaigellae is located only in an extracellular position inside cavities of the mesohyl. Both algae differ from other symbiotic Cyanophyceae in having a normal cell wall. They are able to reproduce in symbiotic condition, but also undergo, in their various extracellular and intracellular positions, a massive process of disintegration. A large amount of algal material is dispersed in the sponge tissues, which is a confirmation, at the ultrastructural level, of trophic relationships in the symbiosis Aphanocapsa-Ircinia.     

Sexual and asexual reproduction in two species of Tethya (Porifera: Demospongiae) from a Mediterranean coastal lagoon

The sexual and asexual phases of reproductive cycles of two sponges, Tethya citrina and T. aurantium, living sympatrically in a Mediterranean coastal lagoon (Stagnone di Marsala, NW Sicily) were studied from samples collected over an 18-mo period. Both species are oviparous and gonochoric. They have a summer, partially overlapping, period of oocyte production, although T. citrina appear to mature earlier. No males were found, possibly due to the very short period of spermatogenesis. Both species produce asexual buds during the autumn/winter months. However, they seem to follow different reproductive strategies, with T. citrina showing a significantly lower production of buds than T. aurantium; by contrast, egg production is significantly lower in the latter species. The difference in the reproductive resource allocation is consistent with data reported in the literature on the anatomy features, genetic population structure and ecological distribution.     

Carotenoid patterns in twenty-nine species of sponges in the order Poecilosclerida (Porifera: Demospongiae): a possible tool for chemosystematics

We assessed the utility of separated carotenoid mixtures for sponge systematics. Carotenoids were extracted from 29 species of 22 genera of 6 families for the demosponge order Poecilosclerida. Mixtures were separated (by thinlayer chromatography on silica gel) without chemical modification and after alteration by saponification, reduction or acetylation or a combination thereof. This approach allowed analysis of individual colonies of less than 1 g wet weight. Relationships among taxa were determined through use of Lawson's similarity index and discriminative analysis. Analysis of multiple runs of fractions from one individual of Ophlitaspongia pennata showed the method to be consistent and reliable. Comparison of specimens of species collected at different localities and times showed high correspondence of carotenoid patterns except in those species suspected of representing species complexes. Analysis of members of the family Clathriidae and of relationships within the order Poecilosclerida suggest that fatty acids esterified to carotenoids may be family-specific and of more taxonomic value than carotenoids. The importance of testing for seasonal, geographic and habitat variability in biochemical patterns is stressed. We discuss the significance of these methods to sponge systematics and their use in comparative studies.     

Biochemical genetic divergence and systematics in sponges of the genera Corticium and Oscarella (Demospongiae: Homoscleromorpha) in the Mediterranean Sea

The sponge sub-class Homoscleromorpha is generally considered to include just two families, the Oscarellidae (without spicules) and the Plakinidae (with simple spicules). In May 1990, an unusual sponge was found deep inside a submarine cave in the western Mediterranean Sea. On the basis of externally visible characters this sponge appeared indistinguishable from the common plakinid species Corticium candelabrum Schmidt, 1862. However, on closer examination in the laboratory the new sponge proved to be devoid of spicules. Therefore, despite great morphological similarities to C. candelabrum, the new sponge should, by taxonomic convention, have been placed in the Oscarellidae. On the basis of other criteria, the similarities to C. candelabrum were great and the new sponge was at first considered to be conspecific. Thus, the taxonomic position of the new sponge and its relationship to C. candelabrum are highly confusing. It could be an aspiculate morph of C. candelabrum, or a new and undescribed related species or, lacking spicules, it could justifiably be placed in a different family (Oscarellidae). The relationship of the new sponge to C. candelabrum and also to two species of Oscarella (Oscarellidae) was assessed by the use of enzyme electrophoresis to estimate genetic divergence between species. It was found that the new sponge was reproductively isolated from sympatric C. candelabrum, with 6 of 16 loci proving diagnostic. Thus it is clear that the new sponge belongs to a different biological species. Surprisingly it was also found that, although this new species was fairly closely related to C. candelabrum (level of genetic identity, I0.47), the two Oscarella species were similarly closely related to C. candelabrum (I0.31 to 0.41) and rather less closely to the new species (I0.17 to 0.28). Indeed from genetic identity estimates, O. tuberculata is more closely related to C. candelabrum than it is to O. lobularis. It is concluded that all homoscleromorph sponges should be placed in the single family Plakinidae.     

Asexual reproduction in homoscleromorph sponges (Porifera; Homoscleromorpha)

Asexual reproduction by external budding in Homoscleromorpha is reported for the first time. Two Mediterranean sponge species were studied, Oscarella lobularis and O. tuberculata. Buds are formed in the marginal basal part of sponge. Budding takes from 1 to 4 days and is defined in three budding stages. First, small irregular protuberances, consisting of external parental tissue, are formed. Second, they elongate and acquire more regular, nipple-like shape. These protuberances are tube like, their internal cavity derived from parental exhalant canal. The wall consists of three layers: (a) external layer is flagellated exopinacoderm, (b) internal one is flagellated endopinacoderm and (c) intermediate one is a thin layer of mesohyl. Third, a spherical bud with a large central cavity is formed. During budding, we did not observe cell proliferation or transdifferentiation either in budding zone or in any special mitotically active region. The bud attached to the substrate is similar to the rhagon developing after larva metamorphosis, it has a syconoid organization. Morphogenetically, budding in Oscarella differes from that in other sponges. Occurring by epithelial morphogenesis, it is similar to morphallaxis during regeneration. The presence in Homoscleromorpha of an epithelial morphogenesis is unique among sponges. This feature is shared by Homoscleromorpha and Eumetazoa.     

Apoptosis in marine sponges: a biomarker for environmental stress (cadmium and bacteria)

The marine demosponge Suberites domuncula is abundantly present on muddy sand bottoms, both in the open sea and in harbors. In the present study it is shown that exposure of S. domuncula to cadmium (CdCl₂) in concentrations ranging from 0.01 to 5.0 g ml⁻¹ for up to 5 d results in apoptotic fragmentation of DNA. Kinetics experiments revealed that after 24 h a significant increase of DNA fragmentation already occurred. Besides cadmium a second stimulus was identified to also cause apoptosis in this species, namely exposure to heat-treated Escherichia coli. In order to support the finding that both cadmium and E. coli induce apoptosis in the sponge, expression of the apoptotic gene MA-3 was studied. The cDNA, SDMA3, was isolated and found to be 2247 nucleotides long. The deduced amino acid sequence (M_r 50 765) shares high similarity with the corresponding mouse molecule. Like the mouse gene, the sponge MA-3 gene undergoes increased expression in response to apoptotic stimuli. While the specimens remained alive after treatment with cadmium, the sponges treated with E. coli died after approximately 12 d. The E. coli-treated animals started to form gemmules 10 to 12 d after addition of the bacteria. Hence, the process of apoptosis in sponges is triggered by two different pathways, one which is initiated by exogenous factors, e.g. heavy metals, and a second one, caused by endogenous factors, which leads to gemmule formation and a shift of the presumably immortal cells to mortal cells. The latter assumption is supported by the finding that during the process of bacteria-induced apoptosis, which results in the death of the specimens, the activity of the telomerase drops. It is concluded that the cells which appear to be immortal and telomerase-positive undergo apoptosis during the process of gemmule formation. In consequence cells not involved in the production of gemmules become mortal. Based on these data, it is proposed that apoptosis is a suitable biomarker in the bioindicator organism S. domuncula to monitor unfavorable environmental conditions, at least in this animal phylum. Received: 10 November 1997 / Accepted: 6 March 1998     

Sessile and non-sessile morphs of Geodia cydonium (Jameson) (Porifera, Demospongiae) in two semi-enclosed Mediterranean bays

Morphological plasticity and ecological aspects of the demosponge Geodia cydonium (Jameson) were studied from seasonal samples collected over 1 year in two semi-enclosed Mediterranean bays of the Southern Italian coast (Marsala lagoon and Porto Cesareo basin). Sponge specimens present two morphs: sessile and non-sessile, both of which showed constant size distribution and density over the studied year. Sessile specimens were larger in size than non-sessile ones. This feature is particularly evident at Porto Cesareo, where these sponges have a more compact skeletal network than at Marsala (evident both in the cortical spicule size and sponge silica content). Sessile specimens adhere to hard rocky substrates (Porto Cesareo) or phanerogam rhizomes (Marsala); non-sessile ones occur on soft bottom areas. Several morphological and structural features of the non-sessile forms differ in the two environments, but the difference in body shape seems to play the most relevant role in enhancing the colonization of incoherent substrates. Indeed, at Marsala, where the large amount of silt and clay determines the occurrence of a markedly reduced anoxic layer just below the surface of the sediment, non-sessile specimens of G. cydonium are fairly spherical and thus able to roll, dragged by slow circular currents. In addition, the usual association with the red alga Rytyphlöea tinctoria, which almost constantly forms a thick and continuous layer around the sponge, allows them to avoid contact with the substrate. The non-sessile specimens from Porto Cesareo inhabit sandy soft bottoms and are flattened. In such an environment, affected by moderate wave turbulence, the flattened shape widens the contact surface between the body and the substrate, thereby reducing the risk of stranding. The evident signs of abrasion, provided by scanning electron microscopy investigations, on both cortical spicules and outermost sponge surface suggest that sponges rub on the bottom. Sediment, epibiontic organisms, and the phanerogam leaves protect this sciaphilous sponge from high solar radiation, allowing the specimens to live in these shallow environments.     

Multiphase redistribution differences of polycyclic aromatic hydrocarbons (PAHs) between two successive sediment suspensions

Successive sediment suspensions often happen in estuary, yet little research has probed into the difference in the release behaviors of organic compounds among different suspensions. This study took polycyclic aromatic hydrocarbons (PAHs) as typical organic contaminants and investigated the release behaviors between two successive suspensions with a particle entrainment simulator (PES). Results showed that successive sediment suspensions lowered the concentration of dissolved PAHs in the overlying water via facilitating the re-adsorption of dissolved PAHs onto the suspended particles. Fast-release and slow-release periods of PAHs were successively observed in the both suspensions. The concentration changes of dissolved PAHs in the second suspension were generally similar with but hysteretic to those in the first suspension. More vigorous desorption and re-absorption of PAHs were induced in the second suspension. Successive sediment suspensions obviously decreased the concentrations of mineral composition and organic matters in the overlying water, which significantly affects multiphase distribution of PAHs.     

The accumulation levels of heavy metals (Ni,Cr, Sr, & Ag) in marine algae from southwest coast of India

In the context of use of marine algae as biological indicators of heavy metal pollution in coastal waters, six species of marine algae collected from the southwest coast of India were analysed for the levels of heavy metals (Ni, Cr, Sr, and Ag). Interspecies and interclass variations were determined on a spatial and temporal scale. The metal contents varied in the ranges, Ni: 0.20–21.06?µg?g^?1 (mean?=?10.13?µg?g^?1), Cr: non-detectable level (ND)–37.18?µg?g^?1 (mean?=?13.86?µg?g^?1), Sr: 2.19–103.90?µg?g^?1 (mean =?29.40?µg?g^?1), and Ag: ND–6.39?µg?g^?1 (mean?=?1.80?µg?g^?1). Ni and Cr contents were of similar magnitude to those reported for algae from polluted areas.     

Trace metals in mussels (Mytilus edulis) from southwest Iceland

The temperate population of the tropical anemonefish Amphiprion clarkii has a seasonality of reproduction, larval settlement and growth. There was a considerable difference in size (15–47 mm) among 0-year olds in December, after the season of larval settlement and growth. This difference could be attributed mainly to a difference in the duration of the first growing season spent after settlement and a social inhibition of growth of the subordinate late settlers by frequent attacks by the dominant early settlers. The growth equation which was used to describe the growth of tagged individuals estimated that an anemonefish would get an opportunity to spawn within four of five years after settlement. Breeding adult pairs occupied almost all sea anemones essential to breeding within a 50×50 m study area, and all 17 new adults that matured from non-breeding juveniles to breeding adults between June 1983 and August 1985 were found in the territories from which either or both of the mated adults had disappeared. The new adults were large juveniles who had resided near those vacant territories before. These results suggest that the onset of breeding by an individual is not only determined by his age, but also by his ranking in the dominance hierarchy. Therefore, the difference in size among 0-year olds in December might give rise to the difference in age of the earliest breeding and might be a factor in making a female apply more reproductive effort at the beginning of the season.     

Resource partitioning by four species of elasmobranchs (Batoidea: Urolophidae) in coastal waters of temperate Australia

Four species of stingarees (Urolophus lobatus, U. paucimaculatus, Trygonoptera personata and T. mucosa) were abundant in samples collected from over sandy substrates in four regions along ∼200 km of the lower west coast of Australia in each season between summer 1990/1991 and winter 1992. The main aim of this study was to determine whether these four species were segregated by habitat, size, sex and/or diet, thereby reducing the potential for intra- and interspecific competition. The densities of U. lobatus and T. personata were almost invariably greater in deep, offshore waters (20 to 35 m) than in shallow, inshore waters (5 to 15 m), while those of U. paucimaculatus were greatest in the southernmost region, irrespective of depth, and those of T. mucosa were highest at the single inshore site at which the water was deep. None of these species was segregated by either size or sex. The volumetric dietary compositions of the two Urolophus species, which consisted predominantly of various crustaceans, and those of the two species of Trygonoptera, which consisted mainly of different polychaetes, were significantly different from each other. The two Urolophus species initially fed largely on amphipods, mysids and carid decapods but, with increasing size, U. lobatus started to ingest teleosts and U. paucimaculatus commenced feeding on errant polychaetes and penaeid decapods. The contribution of errant polychaetes to the diets of T. personata rose progressively from 0% in the smallest fish to 90% in the largest fish, thereby accounting for a concomitant progressive decline in dietary breadth and for intraspecific dietary overlap typically to occur only between successive size classes. In contrast, the volumetric contribution of sedentary polychaetes to the diets of T. mucosa remained high (65 to 80%) throughout life, which accounts for the relatively low and constant dietary breadth of most size classes and a significant overlap in the diets of all but one pair of size classes. The diets of each of the four stingaree species occasionally differed between sites and sometimes underwent seasonal changes, presumably reflecting spatial and temporal differences in the types of prey in the environment. Since the ratios of benthic to epibenthic prey in the diets of the two Trygonoptera species were greater than in those of the two Urolophus species, the first two species apparently forage deeper into the substrate. The combination of partial segregation of the four stingaree species among habitats and the differences in the dietary compositions and feeding strategies of these species, would presumably facilitate the co-existence of these batoid elasmobranchs when they are abundant in the same general region. Received: 29 October 1997 / Accepted: 25 March 1998     

A 3-year investigation of sexual reproduction in <Emphasis Type="Italic">Geodia cydonium</Emphasis> (Jameson 1811) (Porifera,Demospongiae) from a semi-enclosed Mediterranean bay

The reproductive cycle of Geodia cydonium in a semi-enclosed Mediterranean bay (Porto Cesareo, SW Apulia) was studied with monthly frequency over a 3-year period. The investigation was carried out by utilizing a technique consisting of tagging ten individuals with a PVC stick and cutting off, by means of a metal cork borer, small samples (cylinders about 5 cm³ in volume) from each of them for histological analysis. Sexual reproductive elements were detected in all individuals, but in the third year some specimens showed reduced reproductive activity or complete infertility. Spermatogenesis occurred in a short period (from June to August) whereas oogenesis lasted longer (from spring to late summer). Spermatic cysts occurred when the frequency of specimens with oocytes reached its maximum values. A relationship between water temperature and the onset of gamete differentiation was observed. G. cydonium is here confirmed oviparous and gonochoric with a sex ratio in favour of the females. However, in contrast with current literature on Porifera—which suggests that only a limited number of sponges, all belonging to the same species are sexually active—the sexual reproduction of this species involves all the examined individuals. This finding can be explained by the methodological approach used in this research, which differs from the traditional way of assessing the sponge reproductive cycle by analysing randomly collected specimens within a population. The study of a series of individuals over time represents a better “model technique” for investigating sponge sexual reproduction and the effect of environmental parameters on gamete differentiation.     

Density and distribution patterns of the temperate marine fish Cheilodactylus spectabilis (Cheilodactylidae) in a reef environment

Cheilodactylus spectabilis (Hutton) is common over shallow reefs in north-eastern New Zealand. Replicated transect between-area differences in density and in size frequency. C. spectabilis is not nocturnally active. The between-area differences in density and size frequency remained constant over daylight hours, demonstrating that C. spectabilis does not undergo any systematic feeding migrations. Topographic complexity was shown to have a significant positive influence on fish density. Mean size was related to water depth at 5 localities investigated, with small (<200 mm standard length, SL) individuals being restricted to shallow water. All sizes of C. spectabilis examined had fed on small invertebrates, with gammarid amphipods predominating. No evidence of size-related differentiation in feeding patterns was observed, although small individuals spent significantly more time feeding than large ones. Small (<200 mm SL) individuals occupied feeding areas and shelter sites from which they excluded other small C. spectabilis. Large individuals showed no site-associated aggression, covered a greater area during daily movements and overlapped with other large and small C. spectabilis while feeding. No evidence of dawn or dusk peaks of feeding activity or movement was collected.     

Relationships between sponges and crabs: patterns of epibiosis on Inachus aguiarii (Decapoda: Majidae)

The association between sponges and the crab Inachus aguiarii Brito Capello, 1876 was studied by analysing the relationships between sponge distributional patterns on the crab carapaces and several morphological and biological crab parameters. Juveniles, mature females and mature males were differentiated on the basis of sex dimorphism and terminal pubertary moult. All three groups were fouled to different degrees by sponges. Percent sponge cover was related to carapace size only in mature males whereas mature females, all but one of which were ovigerous, were extensively covered regard-less of their size. It is proposed that some behavioural patterns unique to females, such as long resting periods in sponge-rich microhabitats, are responsible for these high sponge covers in females. Sponges showed two trends in the colonization of the carapace, leading to either a monopolizing or a sharing of the available carapace surface. The sponge species found on the carapaces studied are not obligatory epibionts of crabs, but are believed to reflect the sponge population characterizing the crab home range.     

Genetic, morphological, and chemical divergence in the sponge genus Latrunculia (Porifera: Demospongiae) from New Zealand

We undertook a comprehensive study of Latrunculia in New Zealand to determine the relationship between taxonomic, environmental, and chemical variation within the genus. Sponges were collected from five locations around New Zealand: Three Kings Islands, Tutukaka, Wellington, Kaikoura, and Doubtful Sound. Allozyme electrophoresis at nine polymorphic loci indicated that sponges from each geographic location were genetically distinct, and that they displayed genetic differences of the magnitude usually associated with reproductively isolated species (Nei's D between locations =0.375-2.476). Additionally, the comparisons revealed that the green and brown colour morphs of Latrunculia that are sympatric at Three Kings Islands and Kaikoura are distinct from each other, and that there are two genetic groups within the green sponges in Doubtful Sound. On the basis of genetic data we conclude that there are at least eight species of Latrunculia in New Zealand waters, not one to four as had been previously thought. Morphological comparisons of the eight genetic species based on skeletal characters (i.e. skeletal organisation of the choanosome, spicule composition, size, and geometry) indicated that the eight Latrunculia species fell into only two morphological groups that could be easily diagnosed on the basis of discorhabd type. Within these two primary morphological groups, skeletal characteristics among the eight species largely overlap and are not diagnostic. These findings emphasise the limitations of traditional taxonomic methods based solely on skeletal characters for distinguishing species of Latrunculia. However, multivariate analysis (MANOVA and CDA) based on six measured skeletal variables did reveal significant morphological variation among the species (Pillai's Trace=3.28, F=6.90, P<0.0001), supporting the division of the genus into eight species. Comparisons of chemical extracts from Latrunculia also showed that the amounts of five different bioactive compounds (discorhabdins A, B, C, D, and J) varied predictably among the eight species. This finding suggests that discorhabdin variation within Latrunculia, previously thought to be associated with intra-specific environmental variability, is more likely to reflect differences among species rather than phenotypic plasticity. Our results also highlight the importance of thorough taxonomic studies associated with marine natural products research to understand fully the variation in bioactive properties among individuals. The potential processes underlying the unusually high speciation rates in New Zealand Latrunculia that are indicated in our study are discussed.