Anthozoan Species Research Articles

Occurrence of phthalic acid esters (PAEs) and active pharmaceutical ingredients (APIs) in key species of anthozoans in Mediterranean Sea

The Mediterranean Sea's biodiversity is declining due to climate change and human activities, with plastics and emerging contaminants (ECs) posing significant threats. This study assessed phthalic acid esters (PAEs) and active pharmaceutical ingredients (APIs) occurrence in four anthozoan species (Cladocora caespitosa, Eunicella cavolini, Madracis pharensis, Parazoanthus axinellae) using solid phase microextraction (SPME) and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). All specimens were contaminated with at least one contaminant, reaching maximum values of 57.3 ng/g for the ∑PAEs and 64.2 ng/g (wet weight) for ∑APIs, with dibutyl phthalate and Ketoprofen being the most abundant. P. axinellae was the most contaminated species, indicating higher susceptibility to bioaccumulation, while the other three species showed two-fold lower concentrations. Moreover, the potential adverse effects of these contaminants on anthozoans have been discussed. Investigating the impact of PAEs and APIs on these species is crucial, given their key role in the Mediterranean benthic communities.

Basin-scale occurrence and distribution of mesophotic and upper bathyal red coral forests along the Italian coasts

The analysis of 879 ROV dives carried out along the Italian coasts on hard substrata at mesophotic and upper bathyal depths (40-775 m) allowed us to evaluate the current basin-scale presence, relative abundance, bathymetric limits, and habitat preferences of one of the most charismatic Mediterranean habitat-former anthozoan species, Corallium rubrum (Linnaeus, 1758). The species is widespread, and its occurrence ranged from 13% of the explored sites in Ionian Calabria to a hotspot of approximately 80% in Sardinia. Population relative densities were generally low (< 10 colonies m-2), except along the Sardinian coasts and in some areas along the Apulian coast. Almost no red coral colonies were observed between 60 m and 590 m in the nine explored offshore seamounts in the Ligurian and Tyrrhenian Seas. A distinctive coastal distribution discontinuity was found in the Ionian Sea. The optimum bathymetric distribution was between 75 m and 125 m, and no colonies were found below 247 m. Red coral colonies showed a preference for biogenic habitats dominated by crustose coralline algae (CCA) and vertical substrata. The species was absent on iron wrecks. Corallium rubrum disappeared from 14% of the historical fishing banks, while it was confirmed in 86% of them, some of which have been deeply harvested in the past. In particular, the still flourishing Sardinian populations could be supported by the high reproductive potential and favourable hydrodynamic conditions in the area.

The history, biological relevance, and potential applications for polyp bailout in corals.

Corals have evolved a variety of stress responses to changing conditions, many of which have been the subject of scientific research. However, polyp bailout has not received widespread scientific attention, despite being described more than 80 years ago. Polyp bailout is a drastic response to acute stress in which coral colonies break down, with individual and patches of polyps detaching from the colony and the calcareous skeleton Polyps retain their symbiotic partners, have dispersal ability, and may undergo secondary settlement and calcification. Polyp bailout has been described worldwide in a variety of anthozoan species, especially in Scleractinia. It can be induced by multiple natural stressors, but also artificially. Little is known about the evolutionary and ecological potential and consequences of breaking down modularity, the dispersal ability, and reattachment of polyps resulting from polyp bailout. It has been shown that polyp bailout can be used as a model system, with promise for implementation in various research topics. To date, there has been no compilation of knowledge on polyp bailout, which prompted us to review this interesting stress response and provide a basis to discuss research topics and priorities for the future.

Assessment of Romanian circalittoral soft bottom benthic habitats under Danube River influence

Although numerous studies have focused on ecological quality status (EQS) assessments in the Black Sea, knowledge regarding the status of the soft bottom benthic habitats under Danube River influence is still lacking. Therefore, we investigated the diversity, distribution and EQS of benthic habitats located in the northern part of the Romanian continental shelf. In 2018, 34 samples were collected to analyse the macrozoobenthic fauna. A total of 71 species and 6 taxa belonging to 16 phyla were recorded. Two broad-scale circalittoral habitats (1 — terrigenous mud with Melinna palmata; 2 — mixed sediments with Dipolydora quadrilobata and biogenic reefs with Mytilus galloprovincialis) were encountered in the area based on Bray–Curtis similarity index. EQS analysis of the investigated habitats using a new approach based on the reconsideration of the ecological status of invasive species, resulting in the classification of D. quadrilobata as disturbance-tolerant (III) rather than second order opportunistic, allowed us to perform a realistic assessment of the ecological status. The good environmental assessment (GES) - non-GES threshold for AMBI, S, H and M-AMBI*(n) was calculated using a reference site dataset from the period 2014-2018. Overall, the EQS of the analysed habitats showed that 70% of the Dipolydora habitat is in GES according to M-AMBI Dipolydora III and M-AMBI*(n). Fifty percent of the Melinna habitat is in GES according to the same indices. In our study, for the first time on the Romanian coast, we documented the presence of the anthozoan species Edwardsia cf. claparedii.

Cnidarian Immunity and the Repertoire of Defense Mechanisms in Anthozoans.

Anthozoa is the most specious class of the phylum Cnidaria that is phylogenetically basal within the Metazoa. It is an interesting group for studying the evolution of mutualisms and immunity, for despite their morphological simplicity, Anthozoans are unexpectedly immunologically complex, with large genomes and gene families similar to those of the Bilateria. Evidence indicates that the Anthozoan innate immune system is not only involved in the disruption of harmful microorganisms, but is also crucial in structuring tissue-associated microbial communities that are essential components of the cnidarian holobiont and useful to the animal’s health for several functions including metabolism, immune defense, development, and behavior. Here, we report on the current state of the art of Anthozoan immunity. Like other invertebrates, Anthozoans possess immune mechanisms based on self/non-self-recognition. Although lacking adaptive immunity, they use a diverse repertoire of immune receptor signaling pathways (PRRs) to recognize a broad array of conserved microorganism-associated molecular patterns (MAMP). The intracellular signaling cascades lead to gene transcription up to endpoints of release of molecules that kill the pathogens, defend the self by maintaining homeostasis, and modulate the wound repair process. The cells play a fundamental role in immunity, as they display phagocytic activities and secrete mucus, which acts as a physicochemical barrier preventing or slowing down the proliferation of potential invaders. Finally, we describe the current state of knowledge of some immune effectors in Anthozoan species, including the potential role of toxins and the inflammatory response in the Mediterranean Anthozoan Anemonia viridis following injection of various foreign particles differing in type and dimensions, including pathogenetic bacteria.

Ubiquitous macropinocytosis in anthozoans.

Transport of fluids, molecules, nutrients or nanoparticles through coral tissues are poorly documented. Here, we followed the flow of various tracers from the external seawater to within the cells of all tissues in living animals. After entering the general coelenteric cavity, we show that nanoparticles disperse throughout the tissues via the paracellular pathway. Then, the ubiquitous entry gate to within the cells' cytoplasm is macropinocytosis. Most cells form large vesicles of 350-600 nm in diameter at their apical side, continuously internalizing their surrounding medium. Macropinocytosis was confirmed using specific inhibitors of PI3K and actin polymerization. Nanoparticle internalization dynamics is size dependent and differs between tissues. Furthermore, we reveal that macropinocytosis is likely a major endocytic pathway in other anthozoan species. The fact that nearly all cells of an animal are continuously soaking in the environment challenges many aspects of the classical physiology viewpoints acquired from the study of bilaterians.

Coralligenous formations dominated by Eunicella cavolini (Koch, 1887) in the NE Mediterranean: biodiversity and structure

Coralligenous formations are biogenic structures typical of the underwater Mediterranean seascape. Their intricate, multi-layered species assemblages are composed of perennial, long-lived organisms, particularly vulnerable to natural or human-induced disturbances. Despite their high ecological role and conservation value, few studies have addressed the assemblages outside the NW Mediterranean. This is the first quantitative assessment of coralligenous in the N Aegean Sea (NE Mediterranean), specifically focusing at the upper bathymetric limit of assemblages that are dominated by the yellow gorgonian Eunicella cavolini. The number and percent cover of macrobenthic species were studied at depths of 18 to 35 m, using a photoquadrat method. A total of 99 benthic taxa were identified, out of which 89 perennial ones were used to investigate spatial patterns in assemblage structure, composition, and biodiversity. A mean number of 47 perennial taxa were recorded per site, with encrusting coralline algae and sponges being the dominant groups in percent cover and species number, respectively. Across the studied localities, structural complexity and community composition were overall similar, but assemblages presented distinctive differences at the level of sites highlighting the role of local abiotic and anthropogenic factors in the shaping of the coralligenous. Compared to the rest of the Mediterranean, assemblages hosted a similar number of taxa. However, the number and percent cover of erect bryozoans were generally low, while, apart from E. cavolini, other erect anthozoan species were absent. This work provides an important baseline for comparisons and monitoring at a local or Mediterranean scale level.

Evidence for a Large Expansion and Subfunctionalization of Globin Genes in Sea Anemones.

The globin gene superfamily has been well-characterized in vertebrates, however, there has been limited research in early-diverging lineages, such as phylum Cnidaria. This study aimed to identify globin genes in multiple cnidarian lineages, and use bioinformatic approaches to characterize the evolution, structure, and expression of these genes. Phylogenetic analyses and in silico protein predictions showed that all cnidarians have undergone an expansion of globin genes, which likely have a hexacoordinate protein structure. Our protein modeling has also revealed the possibility of a single pentacoordinate globin lineage in anthozoan species. Some cnidarian globin genes displayed tissue and development specific expression with very few orthologous genes similarly expressed across species. Our phylogenetic analyses also revealed that eumetazoan globin genes form a polyphyletic relationship with vertebrate globin genes. Overall, our analyses suggest that a Ngb-like and GbX-like gene were most likely present in the globin gene repertoire for the last common ancestor of eumetazoans. The identification of a large-scale expansion and subfunctionalization of globin genes in actiniarians provides an excellent starting point to further our understanding of the evolution and function of the globin gene superfamily in early-diverging lineages.

Genetically encoded molecular tools for H 2 O 2 imaging and manipulation

Synthetic biology approaches in modern biosciences allow obtaining living systems with new properties by trans-species and even trans-kingdom transfer of molecular machines. Examples are optogenetics and thermogenetics, genome editing by GRISPR-Cas systems, fluorescent proteins and biosensors. After 200 years from discovery of hydrogen peroxide by Thenard, we now use synthetic biology principles to study functions of H2O2 in biological systems. Tools that we have include genetically encoded fluorescent biosensors for H2O2 based on fluorescent proteins from hydrozoan and anthozoan species and bacterial sensing domains, and metabolic engineering tools that are yeast and prokaryotic enzymes with unique capabilities placed in a context of an animal cell. This allow us to precisely monitor and modulate H2O2 levels inside compartments and sub-compartments of the living cells, in a cell- and tissue-specific way. In my talk I will show how antioxidant systems of the cell maintain engineered intracellular H2O2 gradients and define intercompartment H2O2 traffic. I will also give an overview of several new metabolic engineering tools that allow control over key redox parameters in the living cells.

The distribution of reef-dwelling Waminoa flatworms in bays and on capes of Okinawa Island

Waminoa spp. are acoel worms living on the surface of anthozoans. Despite being potential parasites, there is still not much information available regarding the genus’ distribution or host selectivity. The aim of this study was to evaluate the distribution and occurrence of Waminoa on coral reefs along the west coast of Okinawa Main Island (26.5° N, 127.9° E) in southern Japan, comparing sites located on capes with sites located in bays and identifying associated hosts. Scuba surveys were conducted along eight coral reef sites on the west coast of the island, between June and October 2016. Seven out of eight sites showed the presence of Waminoa. Occurrence and associated hosts varied considerably from site to site, and between sites located within bays and those at capes facing the open ocean. Waminoa individuals were significantly more abundant close to the sandy bottom at the lower end of the reef slopes and were always absent in the shallowest 5 m of every site. Waminoa coverage varied depending on the host anthozoan species. No significant differences in coverage were observed within the same host species at different sites. Thus, in Okinawa, the distribution of Waminoa on coral reefs is not random, but has patterns specific to certain anthozoan host species, with the highest occurrences within 10 m in depth from the sandy bottom areas of reefs.

Identification of long non-coding RNAs in two anthozoan species and their possible implications for coral bleaching

Long non-coding RNAs (lncRNAs) have been shown to play regulatory roles in a diverse range of biological processes and are associated with the outcomes of various diseases. The majority of studies about lncRNAs focus on model organisms, with lessened investigation in non-model organisms to date. Herein, we have undertaken an investigation on lncRNA in two zoanthids (cnidarian): Protolpalythoa varibilis and Palythoa caribaeorum. A total of 11,206 and 13,240 lncRNAs were detected in P. variabilis and P. caribaeorum transcriptome, respectively. Comparison using NONCODE database indicated that the majority of these lncRNAs is taxonomically species-restricted with no identifiable orthologs. Even so, we found cases in which short regions of P. caribaeorum’s lncRNAs were similar to vertebrate species’ lncRNAs, and could be associated with lncRNA conserved regulatory functions. Consequently, some high-confidence lncRNA-mRNA interactions were predicted based on such conserved regions, therefore revealing possible involvement of lncRNAs in posttranscriptional processing and regulation in anthozoans. Moreover, investigation of differentially expressed lncRNAs, in healthy colonies and colonial individuals undergoing natural bleaching, indicated that some up-regulated lncRNAs in P. caribaeorum could posttranscriptionally regulate the mRNAs encoding proteins of Ras-mediated signal transduction pathway and components of innate immune-system, which could contribute to the molecular response of coral bleaching.

Expression patterns of sterol transporters NPC1 and NPC2 in the cnidarian-dinoflagellate symbiosis.

The symbiotic interaction between cnidarians (e.g., corals and sea anemones) and photosynthetic dinoflagellates of the genus Symbiodinium is triggered by both host-symbiont recognition processes and metabolic exchange between the 2 partners. The molecular communication is crucial for homeostatic regulation of the symbiosis, both under normal conditions and during stresses that further lead to symbiosis collapse. It is therefore important to identify and fully characterise the key players of this intimate interaction at the symbiotic interface. In this study, we determined the cellular and subcellular localization and expression of the sterol-trafficking Niemann-Pick type C proteins (NPC1 and NPC2) in the symbiotic sea anemones Anemonia viridis and Aiptasia sp. We first established that NPC1 is localised within vesicles in host tissues and to the symbiosome membranes in several anthozoan species. We demonstrated that the canonical NPC2-a protein is mainly expressed in the epidermis, whereas the NPC2-d protein is closely associated with symbiosome membranes. Furthermore, we showed that the expression of the NPC2-d protein is correlated with symbiont presence in healthy symbiotic specimens. As npc2-d is a cnidarian-specific duplicated gene, we hypothesised that it probably arose from a subfunctionalisation process that might result in a gain of function and symbiosis adaptation in anthozoans. Niemann-Pick type C proteins may be key players in a functional symbiosis and be useful tools to study host-symbiont interactions in the anthozoan-dinoflagellate association.

A diverse host thrombospondin-type-1 repeat protein repertoire promotes symbiont colonization during establishment of cnidarian-dinoflagellate symbiosis.

The mutualistic endosymbiosis between cnidarians and dinoflagellates is mediated by complex inter-partner signaling events, where the host cnidarian innate immune system plays a crucial role in recognition and regulation of symbionts. To date, little is known about the diversity of thrombospondin-type-1 repeat (TSR) domain proteins in basal metazoans or their potential role in regulation of cnidarian-dinoflagellate mutualisms. We reveal a large and diverse repertoire of TSR proteins in seven anthozoan species, and show that in the model sea anemone Aiptasia pallida the TSR domain promotes colonization of the host by the symbiotic dinoflagellate Symbiodinium minutum. Blocking TSR domains led to decreased colonization success, while adding exogenous TSRs resulted in a 'super colonization'. Furthermore, gene expression of TSR proteins was highest at early time-points during symbiosis establishment. Our work characterizes the diversity of cnidarian TSR proteins and provides evidence that these proteins play an important role in the establishment of cnidarian-dinoflagellate symbiosis.

Acropora digitiferaEncodes the Largest Known Family of Fluorescent Proteins that Has Persisted during the Evolution ofAcroporaSpecies

Fluorescent proteins (FPs) are well known and broadly used as bio-imaging markers in molecular biology research. Many FP genes were cloned from anthozoan species and it was suggested that multi-copies of these genes are present in their genomes. However, the full complement of FP genes in any single coral species remained unidentified. In this study, we analyzed the FP genes in two stony coral species. FP cDNA sequences from Acropora digitifera and Acropora tenuis revealed the presence of a multi-gene family with an unexpectedly large number of genes, separated into short-/middle-wavelength emission (S/MWE), middle-/long-wavelength emission (M/LWE), and chromoprotein (CP) clades. FP gene copy numbers in the genomes of four A. digitifera colonies were estimated as 16–22 in the S/MWE, 3–6 in the M/LWE, and 8–12 in the CP clades, and, in total, 35, 31, 33, and 33 FP gene copies per individual shown by quantitative PCR. To the best of our knowledge, these are the largest sets of FP genes per genome. The fluorescent light produced by recombinant protein products encoded by the newly isolated genes explained the fluorescent range of live A. digitifera, suggesting that the high copy multi-FP gene family generates coral fluorescence. The functionally diverse multi-FP gene family must have existed in the ancestor of Acropora species, as suggested by molecular phylogenetic and evolutionary analyses. The persistence of a diverse function and high copy number multi-FP gene family may indicate the biological importance of diverse fluorescence emission and light absorption in Acropora species.

Benthic megafauna on steep slopes at the Northern Mid‐Atlantic Ridge

AbstractThe role of small‐scale (<10 km) habitat availability in structuring deep‐sea hard substratum assemblages is poorly understood. Epibenthic megafauna and substratum availability were studied on steep slopes at the Mid‐Atlantic Ridge from May to July 2010 northwest, northeast, southwest and southeast of the Charlie‐Gibbs Fracture Zone (CGFZ; 48–54°N) at between 2095 and 2601 m depth. Megafauna were six times denser north of the CGFZ compared with the south and differences in density were almost entirely driven by sessile fauna. There was no significant difference in habitat availability amongst sites. Rocky substratum made up 48% of the total area surveyed, with individual transects having between 0% and 82% rock. Assemblage structures were different amongst all superstations. The north was dominated by demospongids and hexactinellids, whereas the southern superstations were dominated by anthozoans and hexactinellids. Differences in megafaunal assemblages north and south of the CGFZ primarily reflected variations in demospongid and anthozoan species composition. With 213–1825 individuals·ha−1, and 7–24 species per superstation, hexactinellids were the most species‐rich (36 species) and cosmopolitan taxa at the study site, supporting observations elsewhere along the ridge and in the CGFZ. The absence of significant differences in substrata availability suggested alternative drivers for density or percentage cover. The amount of hard substratum available only limited sessile megafauna density at one transect that was entirely covered with sediments. Species richness was highest for areas with intermediate values of substratum coverage (35–43% rock).

De Novo Assembly and Characterization of Four Anthozoan (Phylum Cnidaria) Transcriptomes.

Many nonmodel species exemplify important biological questions but lack the sequence resources required to study the genes and genomic regions underlying traits of interest. Reef-building corals are famously sensitive to rising seawater temperatures, motivating ongoing research into their stress responses and long-term prospects in a changing climate. A comprehensive understanding of these processes will require extending beyond the sequenced coral genome (Acropora digitifera) to encompass diverse coral species and related anthozoans. Toward that end, we have assembled and annotated reference transcriptomes to develop catalogs of gene sequences for three scleractinian corals (Fungia scutaria, Montastraea cavernosa, Seriatopora hystrix) and a temperate anemone (Anthopleura elegantissima). High-throughput sequencing of cDNA libraries produced ∼20–30 million reads per sample, and de novo assembly of these reads produced ∼75,000–110,000 transcripts from each sample with size distributions (mean ∼1.4 kb, N50 ∼2 kb), comparable to the distribution of gene models from the coral genome (mean ∼1.7 kb, N50 ∼2.2 kb). Each assembly includes matches for more than half the gene models from A. digitifera (54–67%) and many reasonably complete transcripts (∼5300–6700) spanning nearly the entire gene (ortholog hit ratios ≥0.75). The catalogs of gene sequences developed in this study made it possible to identify hundreds to thousands of orthologs across diverse scleractinian species and related taxa. We used these sequences for phylogenetic inference, recovering known relationships and demonstrating superior performance over phylogenetic trees constructed using single mitochondrial loci. The resources developed in this study provide gene sequences and genetic markers for several anthozoan species. To enhance the utility of these resources for the research community, we developed searchable databases enabling researchers to rapidly recover sequences for genes of interest. Our analysis of de novo assembly quality highlights metrics that we expect will be useful for evaluating the relative quality of other de novo transcriptome assemblies. The identification of orthologous sequences and phylogenetic reconstruction demonstrates the feasibility of these methods for clarifying the substantial uncertainties in the existing scleractinian phylogeny.

Demographic responses to warming: reproductive maturity and sex influence vulnerability in an octocoral

Ocean warming, caused by climate change, is critically impacting marine coastal ecosystems. Benthic organisms, such as anthozoans, are increasingly submitted to high temperatures that cause massive mortalities in tropical and temperate seas. To broaden our understanding of their response to thermal stress, we tested the putative role of reproductive maturity and sex in the susceptibility of the Mediterranean red gorgonian, Paramuricea clavata, to high temperatures. We experimentally compared the response to thermal stress of sexually immature (i.e., juveniles) versus mature individuals (i.e., adults), and of males versus females. Colonies' response was firstly assessed by measuring the percentage of tissue area exhibiting necrosis. Then, the reproductive output (i.e., fertility, size, and number of gonads) of both sexes was characterized. When compared to juveniles, adults showed a significantly higher percentage of necrosis, suggesting that during the reproductive period they are more vulner- able to high temperatures. Males and females showed a similar percentage of tissue damage and a significant decrease in their reproductive output. However, females' reproduction was more impacted, suggesting that females are more susceptible to thermal stress than males. A dif- ferential energy investment in reproduction may be the underlying cause of the observed responses. Adults invest a large proportion of their energy budget in reproduction; hence, they have fewer resources available to cope with stress, compared to juveniles. A similar situation seems to apply to females, with respect to males. Considering the current ocean-warming trend, our results imply that the long-term viability of shallow populations of long-lived anthozoans may be jeopardized in the future. This study reveals potential demographic consequences of warming that go beyond its associated increment of mortality rates. Given the important ecological role of many anthozoan species, these results can help better predict the future effects of climate change on coastal ecosystems.

New records of anthozoan species (Cnidaria, Octocorallia, Hexacorallia) for the Turkish Straits System

Abstract This study includes new records of soft corals found in the Turkish Straits System (Dardanelles) between 2013-2014. In the above-mentioned area, 50 stations were searched by scientific divers from the intertidal zone to a depth of over 45 m and the ecological characteristics with substrate differences were also recorded. A total of 47 cnidarians were examined during the surveys. Eleven anthozoan species (Aiptasia mutabilis, Alcyonium acaule, Andresia parthenopea, Calliactis parasitica, Cereus pedunculatus, Condylactis aurantiaca, Epizoanthus couchii, Maasella edwardsi, Pteroeides spinosum, Sarcodictyon roseum, Veretillum cynomorium) belonging to 4 orders are new to the Dardanelles strait, one of which is reported from the Marmara Sea. The current data contributed to the advancement of the knowledge about anthozoan species in the Turkish Seas.

TIR-domain-containing protein repertoire of nine anthozoan species reveals coral–specific expansions and uncharacterized proteins

The intracellular toll/interleukin-1 receptor (TIR) domain plays an important role in vertebrate immunity, but the evolution and function of invertebrate TIR-domain-containing proteins is not fully understood. This study characterized and compared the TIR-domain-containing protein repertoire of nine cnidarians in class Anthozoa. A diverse set of proteins, including MyD88 (myeloid differentiation primary response protein 88), toll-like receptor (TLR)-like, interleukin-1 receptor (IL-1R)-like, and TIR-only proteins are present in the species surveyed. Increased numbers of TIR-only proteins were observed in corals compared to anemones, especially in the Acroporid and Pocilloporid coral families. This expansion could be linked to diversity of the microbial community on or in hosts and managing both positive and negative associations. Phylogenetic analysis indicates there are two groups of proteins with IL-1R-like domain architecture in anthozoans that potentially evolved independently of the vertebrate family. Bacterial-like TIR_2 domain proteins are also present, including one sequence with novel domain architecture. Overall this work promotes a better understanding of the anthozoan immune repertoire, which is important in the context learning about ancestral immune pathways and host–microbe interactions.

Mixed medusivory by the sea anemone Entacmaea medusivora (Anthozoa: Actiniaria) in Kakaban Lake, Indonesia

Anchialine lakes are landlocked marine water bodies, which may contain swarms of scyphozoan jellyfish (Becking et al. 2011). Because most jellyfish are pelagic, published examples of their predation by sessile benthic organisms are rare and, in the case of anthozoan predators, only single prey species have been clearly documented (Fautin and Fitt 1991; Jarms and Tiemann 2004; Alamaru et al. 2009). Two anchialine lakes are known to contain the medusivorous sea anemone Entacmaea medusivora Fautin and Fitt 1991: Jellyfish Lake in Palau (Fautin and Fitt 1991) andKakaban Lake in East Kalimantan, Indonesia (Tomascik and Mah 1994). In Palau, this species preys on Mastigias papua (Lesson, 1830) (Fautin and Fitt 1991). During surveys of Kakaban Lake (02°08′27′′N, 118°30′40′′ E) in October 2003 and August 2008, medusivory by E. medusivora was observed at <3 m depth. Sea anemones occurred on substrates consisting of green alga, Halimeda sp. (Fig. 1a, b), logs, and mangrove roots. Three species of jellyfish acted as prey: Mastigias papua (Fig. 1c, d), Aurelia aurita (Linnaeus, 1758) (Fig. 1e, f), and Cassiopeia ornata Haeckel 1880 (Fig. 1g, h). This study is the first to present in situ photographic evidence of a benthic anthozoan species predating on more than one species of Scyphozoa. Experiments have