Ainara Ballesteros

The Mediterranean Sea, although a biodiversity hotspot, is one of the most affected seas by non-indigenous species (NIS). This problem is worsened by rising sea temperatures due to climate change, which promotes the spread of thermophilic species. Among the NIS scyphozoan jellyfish species recorded in the Mediterranean, Cassiopea andromeda – commonly known as the “upside-down jellyfish”– is a notable example. Observadores del Mar ( OdM ) is the leading platform for marine citizen science in Spain and works towards ocean conservation and health. It is a well-established tool for generating knowledge in marine research and has successfully provided early warning of NIS reports in the Mediterranean, while also serving as an effective network for the monitoring of NIS and other indicators. Three reports of C. andromeda from Almeria, southern Spain, have been reported in OdM and thanks to the involvement of its community, 12 samples were collected for phylogenetic analysis and monitoring was done for 15 months in the study area. The results confirmed the first record of C. andromeda in Spanish Mediterranean waters representing the westernmost record in the basin. Monitoring also suggests the species establishment in the area. This study contributes to the knowledge of C. andromeda invasiveness and highlights the importance of marine citizen science in the detection and monitoring of NIS. It also underscores the collaboration and commitment already established between scientists and citizens, which will allow further progress in the fields of biological invasions, management, and policy.

Rhizostoma pulmo is a widely distributed scyphozoan in the Mediterranean Sea. Their stings result mainly in erythema, small vesicles, or/and pain, and cause a high number of bathers to seek assistance from first-aid services during the summer season. Despite the threat that jellyfish stings represent to public health, there is disagreement in the scientific community on first-aid protocols, with the dispute largely centered around the effectiveness of vinegar. In the present research, we investigated the effect of commonly used rinse solutions on nematocyst discharge in R. pulmo and the effect of vinegar on three more scyphozoans (Aurelia sp., Cassiopea sp., and Rhizostoma luteum). Scented ammonia, vinegar, and acetic acid triggered nematocyst discharge in R. pulmo. Vinegar also caused nematocyst discharge in Aurelia sp., Cassiopea sp., and R. luteum. In contrast, seawater, baking soda, freshwater, urine, and hydrogen peroxide were considered neutral solutions that did not induce nematocyst discharge. These results indicate that the use of vinegar, acetic acid, or commercial products based on these compounds is counterproductive. Their use can worsen pain and discomfort caused not only by R. pulmo stings but also by those of any scyphozoan. The use of seawater is recommended for cleaning the R. pulmo sting site until an inhibitor solution that irreversibly prevents nematocyst discharge is discovered.

Pelagia noctiluca stings are common in Mediterranean coastal areas and, although the venom is non-lethal, they are painful. Due to its high toxicity and abundance, P. noctiluca is considered a target species for the focus of research on active ingredients to reduce the symptoms of its sting. To determine the effect of 31 substances and formulations on nematocyst discharge, we performed three tests: (1) screening of per se discharge activator solutions, (2) inhibitory test with nematocyst chemical stimulation (5% acetic acid) and (3) inhibitory test quantifying the hemolytic area. Ammonia, barium chloride, bleach, scented ammonia, carbonated cola, lemon juice, sodium chloride and papain triggered nematocyst discharge. All of them were ruled out as potential inhibitors. Butylene glycol showed a reduction in nematocyst discharge, while the formulations of 10% lidocaine in ethanol, 1.5% hydroxyacetophenone in distilled water + butylene glycol, and 3% Symsitive^® in butylene glycol inhibited nematocyst discharge. These last results were subsequently correlated with a significant decrease in hemolytic area in the venom assays versus seawater, a neutral solution. The presented data represent a first step in research to develop preventive products for jellyfish stings while at the same time attempting to clarify some uncertainties about the role of various topical solutions in P. noctiluca first-aid protocols.

Interest in the rearing of jellyfish has grown exponentially over recent years due to their indisputable potential in a wide variety of uses and research. Here, we describe the design and operation of an effective modified kreisel aquarium that allows to grow of the early planktonic life stages of jellyfish in both flow-through or closed systems. Thanks to its versatile and innovative approach, the aquarium operates for species with a metagenetic life cycle, and allows the metamorphosis of ephyrae from fertilized eggs for species with a holoplanktonic life cycle, such as Pelagia noctiluca. In addition, its configuration allows the mesh screen to be changed in situ, adjusting the size of the mesh according to the growth of the jellyfish and the size of the prey offered. An accessory, named the polyp plate, suspends the polyps in a downward or sideways position, facilitating prey capture, strobilae formation and release of ephyrae. The improvements of this modified kreisel reduce the time-involvement for staff in maintenance routines, and it is an important contribution to jellyfish husbandry techniques and biomass production. These improvements especially target to the culture of the blooming and stinging P. noctiluca, which has been noted to present a host of challenges to the scientific and aquarist communities.

The scyphozoan Pelagia noctiluca has potential value for research and a wide variety of industrial applications. However, its holoplanktonic life cycle makes it difficult to maintain a permanent stock in aquaculture, unlike the continuous supply of newly-released ephyrae from jellyfish species with a benthic life stage. We investigated optimal feeding conditions for the captive breeding of P. noctiluca, monitoring total body diameter, morphological development, survival rate, and instantaneous growth rate. Diets of different items encapsulated in gelatin or non-motile prey were not suitable for successful growth of the early stages (ephyra and metaephyra), whereas rotifers were a sufficient food source for healthy growth. Fertilized eggs from sexually-mature jellyfish were obtained at 84.5 ± 2.1 and 87.0 ± 9.9 days post-fecundation in the presence and absence of gelatinous prey (Aurelia sp.), respectively. In contrast with the general consensus, we demonstrate that the incorporation of medusivorous prey was not essential for proper growth and closure of P. noctiluca life cycle. Instead, we recommend the inclusion of white shrimps (Acetes sp.) for the formation and maturation of gonads and production of eggs. Instantaneous growth rates (57.0% d^-1), the highest reported to date for this species in captive conditions, were higher during the early stages. A third generation of cultured P. noctiluca was obtained for the first time (169 – 179 experimental days), demonstrating the durability of the culture, providing permanent stock and halving the production time of mature individuals compared to previous studies. Finally, we validated the use of captive-bred specimens through quantification of the hemolytic area after the sting process using the tentacle skin blood agarose assay methodology.

Anemonia viridis is an abundant and widely distributed temperate sea anemone that can form dense congregations of individuals. Despite the potential severity of its sting, few detailed cases have been reported. We report a case of a severe toxic reaction following an A. viridis sting in a 35-year-old oceanographer. She developed severe pain, itching, redness, and burning sensation, which worsened one week after treatment with anti-inflammatories, antihistamines and corticosteroids. Prompted by this event, and due to the insufficient risk prevention, lack of training for marine-environment users, and lack of research into sting-specific first-aid protocols, we evaluated the cnidocyst response to five different compounds commonly recommended as rinse solutions in first-aid protocols (seawater, vinegar, ammonia, baking soda, and freshwater) by means of the Tentacle Solution Assay. Vinegar and ammonia triggered an immediate and massive cnidocyst discharge after their application and were classified as activator solutions. Baking soda and freshwater were also classified as activator solutions, although with a lower intensity of discharge. Only seawater was classified as a neutral solution and therefore recommended as a rinse solution after A. viridis sting, at least until an inhibitory solution is discovered.

Anemonia viridis is an abundant and widely-distributed temperate sea anemone that can form dense congregations of individuals. Despite the potential severity of its sting, few detailed cases have been reported. We report a case of a severe toxic reaction following an A. viridis sting in a 35-year-old oceanographer. She developed severe pain, itching, redness and burning sensation, which worsened one week after treatment with anti-inflammatories, antihistamines and corticosteroids. Prompted by this event, and due to the insufficient risk prevention, lack of training for marine-environment users and lack of research into sting-specific first-aid protocols, we evaluated the cnidocyst response to five different compounds commonly recommended as rinse solutions in first-aid protocols (seawater, vinegar, ammonia, baking soda and freshwater) by means of the Tentacle Solution Assay. Vinegar and ammonia triggered an immediate and massive cnidocyst discharge after their application and were classified as activator solutions. Baking soda and freshwater were also classified as activator solutions, although with a lower intensity of discharge. Only seawater was classified as a neutral solution and therefore recommended as a rinse solution after A. viridis sting, at least until an inhibitory solution is discovered.

Pelagia noctiluca is considered the most important jellyfish in the Mediterranean Sea, due to its abundance and the severity of its stings. Despite its importance in marine ecosystems and the health problems caused by its massive arrival in coastal areas, little is known about its early life stages and its cnidome has never been described. This study of the morphological and anatomical features throughout the life cycle identifies four early stages: two ephyra and two metaephyra stages. Ephyra stage 1, newly developed from a planula, has no velar canals, gastric filaments or nematocyst batteries. Ephyra stage 2, has velar canals, a cruciform-shaped manubrium and gastric filaments. Metaephyra stage 3 has eight tentacle buds and nematocyst clusters for the first time. Lastly, in metaephyra stage 4, the eight primary tentacles grow nearly simultaneously, with no secondary tentacles. Complete nematocyst battery patterns gradually develop throughout the later life stages. Four nematocyst types are identified: a-isorhiza, A-isorhiza, O-isorhiza and eurytele. Of these, a-isorhiza and eurytele are the most important throughout the entire life cycle, while A-isorhiza and O-isorhiza have a more important role in advanced stages. All nematocysts show a positive correlation between increasing capsule volumes and increasing body diameter of the ephyrae, metaephyrae, young medusae and adult medusae. In the early stages, the volumes of euryteles in the gastric filaments are larger than those in the exumbrella, indicating that the capsule volume is critical in the absence of marginal tentacles, specialized for feeding. This study provides updated information, the most extensive description to date, including high-resolution photographs and schematic drawings of all the developmental stages in the life cycle of P. noctiluca. Additionally, the first cnidome characterization is provided for each stage to facilitate accurate identification of this species when collected in the water column, and to raise awareness of the potential for human envenomation.

The jellyfish species that inhabit the Mediterranean coastal waters are not lethal, but their stings can cause severe pain and systemic effects that pose a health risk to humans. Despite the frequent occurrence of jellyfish stings, currently no consensus exists among the scientific community regarding the most appropriate first-aid protocol. Over the years, several different rinse solutions have been proposed. Vinegar, or acetic acid, is one of the most established of these solutions, with efficacy data published. We investigated the effect of vinegar and seawater on the nematocyst discharge process in two species representative of the Mediterranean region: Pelagia noctiluca (Scyphozoa) and Carybdea marsupialis (Cubozoa), by means of (1) direct observation of nematocyst discharge on light microscopy (tentacle solution assay) and (2) quantification of hemolytic area (tentacle skin blood agarose assay). In both species, nematocyst discharge was not stimulated by seawater, which was classified as a neutral solution. In P. noctiluca, vinegar produced nematocyst discharge per se, but inhibited nematocyst discharge from C. marsupialis. These results suggest that the use of vinegar cannot be universally recommended. Whereas in case of a cubozoan C. marsupialis sting, the inhibitory effect of vinegar makes it the ideal rinse solution, in case of a scyphozoan P. noctiluca sting, vinegar application may be counterproductive, worsening the pain and discomfort of the stung area.

Although the jellyfish species that inhabit the Mediterranean coastal waters are not lethal, their sting can cause severe pain and systemic effects that pose a health risk to humans. Despite the frequent occurrence of jellyfish stings, currently no consensus exists regarding the most appropriate first aid protocol among the scientific community. Over the years, several different rinse solutions have been proposed of which vinegar, or acetic acid, is one of the most noticeable as a rinse solution with efficacy data published. We investigated the effect of vinegar and seawater on the nematocyst discharge process in species representative of the Mediterranean region such as Pelagia noctiluca (Scyphozoa) and Carybdea marsupialis (Cubozoa) by means of: (1) direct observation on the nematocyst discharge by light microscope (Tentacle solution assay) and (2) the quantification of hemolytic area (Tentacle skin blood agarose assay). The nematocyst discharge of both species was not stimulated by seawater, and it was classified as a neutral solution. In P. noctiluca, vinegar produced nematocyst discharge per ser while inhibiting the nematocyst discharge of C. marsupilais. These results suggest that the use of vinegar cannot be universally recommended. While in case of a cubozoan C. marsupialis sting, the inhibitory effect of vinegar makes it the best rinse solution par excellence, in case of a scyphozoan P. noctiluca sting, its application can be counterproductive, worsening the pain and discomfort of the stung area.