A new transcriptome and transcriptome profiling of adult and larval tissue in the box jellyfish Alatina alata: an emerging model for studying venom, vision and sex.

BACKGROUND: Cubozoans (box jellyfish) are cnidarians that have evolved a number of distinguishing features. Many cubozoans have a particularly potent sting, effected by stinging structures called nematocysts; cubozoans have well-developed light sensation, possessing both image-forming lens eyes and light-sensitive eye spots; and some cubozoans have complex mating behaviors, including aggregations, copulation and internal fertilization. The cubozoan Alatina alata is emerging as a cnidarian model because it forms predictable monthly nearshore breeding aggregations in tropical to subtropical waters worldwide, making both adult and larval material reliably accessible. To develop resources for A. alata, this study generated a functionally annotated transcriptome of adult and larval tissue, applying preliminary differential expression analyses to identify candidate genes involved in nematogenesis and venom production, vision and extraocular sensory perception, and sexual reproduction, which for brevity we refer to as "venom", "vision" and "sex". RESULTS: We assembled a transcriptome de novo from RNA-Seq data pooled from multiple body parts (gastric cirri, ovaries, tentacle (with pedalium base) and rhopalium) of an adult female A. alata medusa and larval planulae. Our transcriptome comprises ~32 K transcripts, after filtering, and provides a basis for analyzing patterns of gene expression in adult and larval box jellyfish tissues. Furthermore, we annotated a large set of candidate genes putatively involved in venom, vision and sex, providing an initial molecular characterization of these complex features in cubozoans. Expression profiles and gene tree reconstruction provided a number of preliminary insights into the putative sites of nematogenesis and venom production, regions of phototransduction activity and fertilization dynamics in A. alata. CONCLUSIONS: Our Alatina alata transcriptome significantly adds to the genomic resources for this emerging cubozoan model. This study provides the first annotated transcriptome from multiple tissues of a cubozoan focusing on both the adult and larvae. Our approach of using multiple body parts and life stages to generate this transcriptome effectively identified a broad range of candidate genes for the further study of coordinated processes associated with venom, vision and sex. This new genomic resource and the candidate gene dataset are valuable for further investigating the evolution of distinctive features of cubozoans, and of cnidarians more broadly.

Use of an Inverse Method for Time Series to Estimate the Dynamics of and Management Strategies for the Box Jellyfish Carybdea marsupialis.

Frequently, population ecology of marine organisms uses a descriptive approach in which their sizes and densities are plotted over time. This approach has limited usefulness for design strategies in management or modelling different scenarios. Population projection matrix models are among the most widely used tools in ecology. Unfortunately, for the majority of pelagic marine organisms, it is difficult to mark individuals and follow them over time to determine their vital rates and built a population projection matrix model. Nevertheless, it is possible to get time-series data to calculate size structure and densities of each size, in order to determine the matrix parameters. This approach is known as a "demographic inverse problem" and it is based on quadratic programming methods, but it has rarely been used on aquatic organisms. We used unpublished field data of a population of cubomedusae Carybdea marsupialis to construct a population projection matrix model and compare two different management strategies to lower population to values before year 2008 when there was no significant interaction with bathers. Those strategies were by direct removal of medusae and by reducing prey. Our results showed that removal of jellyfish from all size classes was more effective than removing only juveniles or adults. When reducing prey, the highest efficiency to lower the C. marsupialis population occurred when prey depletion affected prey of all medusae sizes. Our model fit well with the field data and may serve to design an efficient management strategy or build hypothetical scenarios such as removal of individuals or reducing prey. TThis This sdfsdshis method is applicable to other marine or terrestrial species, for which density and population structure over time are available.

Monodisc strobilation in Japanese giant box jellyfish Morbakka virulenta (Kishinouye, 1910): a strong implication of phylogenetic similarity between Cubozoa and Scyphozoa.

Both sexes of the Japanese giant box jellyfish Morbakka virulenta were collected from the Seto Inland Sea, western Japan in December 2011, in order to observe the developmental processes from polyps to medusae. The medusa production in M. virulenta is up to now a unique process in cubozoans in that it exhibits a form of monodisc strobilation where the polyp is regenerated before the medusa detaches. This mode of medusa production was previously thought to be exclusive to scyphozoans. The general shape of young medusae resembles that of other cubozoans such as Alatina moseri and Copula sivickisi, but is differentiated from these by the short capitate tentacles and the lack of gastric filaments in the stomach. The unique medusa production of M. virulenta highly implies a phylogenetic similarity between cubozoans and scyphozoans.

Cell proliferation in cubozoan jellyfish Tripedalia cystophora and Alatina moseri.

Cubozoans (box jellyfish) undergo remarkable body reorganization throughout their life cycle when, first, they metamorphose from swimming larvae to sessile polyps, and second, through the metamorphosis from sessile polyps to free swimming medusae. In the latter they develop complex structures like the central nervous system (CNS) and visual organs. In the present study several aspects of cell proliferation at different stages of the life cycle of the box jellyfish Tripedalia cystophora and Alatina moseri have been examined through in vivo labeling of cells in the synthetic phase (S phase) of the cell cycle. Proliferation zones were found in metamorphosing polyps, as well as in juvenile medusae, where both the rhopalia and pedalia have enhanced rates of proliferation. The results also indicate a rather fast cell turnover in the rhopalia including the rhopalial nervous system (RNS). Moreover, T. cystophora showed diurnal pattern of cell proliferation in certain body parts of the medusa, with higher proliferation rates at nighttime. This is true for two areas in close connection with the CNS: the stalk base and the rhopalia.

Early life history of Alatina cf. moseri populations from Australia and Hawaii with implications for taxonomy (Cubozoa: Carybdeida, Alatinidae).

The early life stages of the cubomedusa Alatina cf. moseri from Osprey Reef (North Queensland, Australia) and Waikiki (Oahu, Hawaii) were studied using laboratory-based culturing conditions. Spawning populations from both regions were observed with reliable periodicity allowing polyp cultures from these locations to be collected and established under laboratory conditions. The polyps of this species were successfully reared from spawning adults. Polyps of Alatina cf. moseri were cultured at temperatures of 23-28 °C, developed up to 19 tentacles and reached up to 1.70 mm in height. The balloon-shaped hypostomes possessed 4 well-defined lips. The polyps increased their numbers by means of formation of either sedentary polyp buds or creeping-polyp buds, which attached after 2-3 days. Metamorphosis occurred at temperatures of 25-28 °C. Development of polyps and medusae were achieved for the first time within the genus Alatina and allowed comparisons of early life history between these and other species of the Carybdeida families. The metamorphosis and young medusa of this genus showed characters that differed distinctly from those noted for other Carybdeida species, but are very similar to the one described from Puerto Rico by Arneson and Cutress in 1976 for Alatina sp. (named by them Carybdea alata). Based on this evidence, the discrepancies in original specimen descriptions and the previous genetic comparisons, we support the suggestion that the two previously described species of Alatina from Australia and Hawaii (Alatina mordens and Alatina moseri) appear to represent artificial taxonomic units and may in fact be the same as the original Carybdea alata species named from Puerto Rico. Further taxonomic studies are desperately needed in order to clarify the various species and description discrepancies that exist within this newly proposed genus.

Seasonality in polyps of a tropical cubozoan: A latina nr mordens.

A latina nr mordens have been located in large predictable spawning aggregations near Osprey Reef in the Coral Sea eight to ten days after a full moon; however, polyps have never been located in-situ. The polyp stage contributes to the abundance of medusae through asexual reproduction and metamorphosis, and may influence the periodicity of medusae by metamorphosis of the polyp. To elucidate the relationship between medusae periodicity and polyp ecology, polyps were exposed to thermal and osmotic treatments in order to determine the theoretical environmental limits to their distribution. Maximum fecundity occurred in thermal treatments of 21 to 25ºC and the theoretical minimum thermal requirement for population stability was approximately 17ºC. Polyps were also exposed to five feeding regimes and fecundity was found to be positively correlated with feeding frequency. Thermal and osmotic variations did not induce metamorphosis in this species, however, reduced food did. The implications of asexual reproduction and cues for metamorphosis in relation to population dynamics of this species are discussed.

Growth, development and temporal variation in the onset of six Chironex fleckeri medusae seasons: a contribution to understanding jellyfish ecology.

Despite the worldwide distribution, toxicity and commercial, industrial and medical impacts jellyfish present, many aspects of their ecology remain poorly understood. Quantified here are important ecological parameters of Chironex fleckeri medusae, contributing not only to the understanding of an understudied taxon, the cubozoa, but also to the broader understanding of jellyfish ecology. C. fleckeri medusae were collected across seven seasons (1999, 2000, 2003, 2005-07 and 2010), with growth rates, temporal variation in the medusae season onset and differences in population structure between estuarine and coastal habitats quantified. With a mean of 2 September ± 2 d (mean ± 95% confidence limits), the earliest date of metamorphosis was temporally constrained between seasons, varying by only 7 d (30 August to 5 September). Juvenile medusae appeared to be added over an extended period, suggesting polyp metamorphosis was an ongoing process once it commenced. At a maximum of 3 ± 0.2 mm d(-1) IPD, medusae growth to an asymptotic size of ~190 mm IPD was rapid, yet, with the oldest medusae estimated to be ~78 d in age, medusae did not appear to accumulate along the coastline. Furthermore, a greater proportion of juveniles were observed along the coastline, with estuarine populations typified by larger medusae. With key aspects of C. fleckeri's ecology now quantified, medusae season management protocols can be further developed.

Evolution of box jellyfish (Cnidaria: Cubozoa), a group of highly toxic invertebrates.

Cubozoa (Cnidaria: Medusozoa) represents a small clade of approximately 50 described species, some of which cause serious human envenomations. Our understanding of the evolutionary history of Cubozoa has been limited by the lack of a sound phylogenetic hypothesis for the group. Here, we present a comprehensive cubozoan phylogeny based on ribosomal genes coding for near-complete nuclear 18S (small subunit) and 28S (large subunit) and partial mitochondrial 16S. We discuss the implications of this phylogeny for our understanding of cubozoan venom evolution, biogeography and life-history evolution. Our phylogenetic hypothesis suggests that: (i) the last common ancestor of Carybdeida probably possessed the mechanism(s) underlying Irukandji syndrome, (ii) deep divergences between Atlantic and Indo-Pacific clades may be explained by ancient vicariant events, and (iii) sexual dimorphism evolved a single time in concert with complex sexual behaviour. Furthermore, several cubozoan taxa are either para- or polyphyletic, and we address some of these taxonomic issues by designating a new family, Carukiidae, a new genus, Copula, and by redefining the families Tamoyidae and Tripedaliidae. Lastly, cubozoan species identities have long been misunderstood and the data presented here support many of the recent scientific descriptions of cubozoan species. However, the results of a phylogeographic analysis of Alatina moseri from Hawai'i and Alatina mordens from Australia indicate that these two nominal species represent a single species that has maintained metapopulation cohesion by natural or anthropogenic dispersal.

Assembly of the cnidarian camera-type eye from vertebrate-like components.

Animal eyes are morphologically diverse. Their assembly, however, always relies on the same basic principle, i.e., photoreceptors located in the vicinity of dark shielding pigment. Cnidaria as the likely sister group to the Bilateria are the earliest branching phylum with a well developed visual system. Here, we show that camera-type eyes of the cubozoan jellyfish, Tripedalia cystophora, use genetic building blocks typical of vertebrate eyes, namely, a ciliary phototransduction cascade and melanogenic pathway. Our findings indicative of parallelism provide an insight into eye evolution. Combined, the available data favor the possibility that vertebrate and cubozoan eyes arose by independent recruitment of orthologous genes during evolution.

Venom ontogeny, diet and morphology in Carukia barnesi, a species of Australian box jellyfish that causes Irukandji syndrome.

Venom profiles of two age groups of the medically important Australian box jellyfish Carukia barnesi [Southcott, R.V., 1967. Revision of some Carybdeidae (Scyphozoa, Cubomedusae), including description of jellyfish responsible for the 'Irukandji' syndrome. Aust. J. Zool. 15, 651-657] were compared. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis revealed differences in protein banding of tentacular venom between immature and mature animals. This correlates to a change in diet from invertebrate prey in immature C. barnesi medusae to vertebrate prey in mature medusae. Unlike other cubozoan studies, a change in venom did not equate to a change in nematocyst types or their relative frequencies. Additionally, comparison of tentacle structure and bell wart number showed developmental differences between the two age classes. Observations of prey capture in mature individuals and differences in bell warts between immature and mature medusae suggest different methods of prey capture are employed at different life stages of C. barnesi.

Australian venomous jellyfish, envenomation syndromes, toxins and therapy.

The seas and oceans around Australia harbour numerous venomous jellyfish. Chironex fleckeri, the box jellyfish, is the most lethal causing rapid cardiorespiratory depression and although its venom has been characterised, its toxins remain to be identified. A moderately effective antivenom exists which is also partially effective against another chirodropid, Chiropsalmus sp. Numerous carybdeids, some unidentified, cause less severe illness, including Carybdea rastoni whose toxins CrTX-A and CrTX-B are large proteins. Carukia barnesi, another small carybdeid is one cause of the 'Irukandji' syndrome which includes delayed pain from severe muscle cramping, vomiting, anxiety, restlessness, sweating and prostration, and occasionally severe hypertension and acute cardiac failure. The syndrome is in part caused by release of catecholamines but the cause of heart failure is undefined. The venom contains a sodium channel modulator. Two species of Physalia are present and although one is potentially lethal, has not caused death in Australian waters. Other significant genera of jellyfish include Tamoya, Pelagia, Cyanea, Aurelia and Chyrosaora.

A survey of the Scyphozoa and Cubozoa (Cnidaria, Medusozoa) from the Ceará coast (NE Brazil) / Levantamento de Scyphozoa e Cubozoa (Cnidaria, Medusozoa) do litoral do Ceará (NE do Brasil)

Most of the scyphozoans and cubozoans recorded for the Brazilian coast are presumed to be distributed over the entire coastline, but little information on precise records is available. This paper presents the first published records for the Ceará state (northeastern region) of the cubozoan Chiropsalmus quadrumanus, and the scyphozoans Chrysaora lactea, Linuche unguiculata (polyp), Lychnorhiza lucerna, Nausithoe sp. (polyp), Phyllorhiza punctata, and Stomolophus meleagris. The medusoid species have been known by local fishermen and non-specialists for a long time in the area.

A maioria das cifomedusas e cubomedusas registradas para o litoral brasileiro é presumivelmente distribuída por toda a costa; porém poucos registros precisos estão disponíveis. Este estudo apresenta os primeiros registros publicados para o estado do Ceará (região Nordeste) da cubomedusa Chiropsalmus quadrumanus, e dos cifozoários: Chrysaora lactea, Linuche unguiculata (pólipo), Lychnorhiza lucerna, Nausithoe sp. (pólipo), Phyllorhiza punctata and Stomolophus meleagris. As espécies de medusas são conhecidas de longa data por pescadores locais e não-especialistas no grupo.