Localization and Bioreactivity of Cysteine-Rich Secretions in the Marine Gastropod Nucella lapillus.

Marine biodiversity has been yielding promising novel bioproducts from venomous animals. Despite the auspices of conotoxins, which originated the paradigmatic painkiller Prialt, the biotechnological potential of gastropod venoms remains to be explored. Marine bioprospecting is expanding towards temperate species like the dogwhelk Nucella lapillus, which is suspected to secrete immobilizing agents through its salivary glands with a relaxing effect on the musculature of its preferential prey, Mytilus sp. This work focused on detecting, localizing, and testing the bioreactivity of cysteine-rich proteins and peptides, whose presence is a signature of animal venoms and poisons. The highest content of thiols was found in crude protein extracts from the digestive gland, which is associated with digestion, followed by the peribuccal mass, where the salivary glands are located. Conversely, the foot and siphon (which the gastropod uses for feeding) are not the main organs involved in toxin secretion. Ex vivo bioassays with Mytilus gill tissue disclosed the differential bioreactivity of crude protein extracts. Secretions from the digestive gland and peribuccal mass caused the most significant molecular damage, with evidence for the induction of apoptosis. These early findings indicate that salivary glands are a promising target for the extraction and characterization of bioactive cysteine-rich proteinaceous toxins from the species.

A Transcriptomic Approach to the Metabolism of Tetrapyrrolic Photosensitizers in a Marine Annelid.

The past decade has seen growing interest in marine natural pigments for biotechnological applications. One of the most abundant classes of biological pigments is the tetrapyrroles, which are prized targets due their photodynamic properties; porphyrins are the best known examples of this group. Many animal porphyrinoids and other tetrapyrroles are produced through heme metabolic pathways, the best known of which are the bile pigments biliverdin and bilirubin. Eulalia is a marine Polychaeta characterized by its bright green coloration resulting from a remarkably wide range of greenish and yellowish tetrapyrroles, some of which have promising photodynamic properties. The present study combined metabolomics based on HPLC-DAD with RNA-seq transcriptomics to investigate the molecular pathways of porphyrinoid metabolism by comparing the worm's proboscis and epidermis, which display distinct pigmentation patterns. The results showed that pigments are endogenous and seemingly heme-derived. The worm possesses homologs in both organs for genes encoding enzymes involved in heme metabolism such as ALAD, FECH, UROS, and PPOX. However, the findings also indicate that variants of the canonical enzymes of the heme biosynthesis pathway can be species- and organ-specific. These differences between molecular networks contribute to explain not only the differential pigmentation patterns between organs, but also the worm's variety of novel endogenous tetrapyrrolic compounds.

Light-Mediated Toxicity of Porphyrin-Like Pigments from a Marine Polychaeta.

Porphyrins and derivatives form one of the most abundant classes of biochromes. They result from the breakdown of heme and have crucial physiological functions. Bilins are well-known representatives of this group that, besides significant antioxidant and anti-mutagenic properties, are also photosensitizers for photodynamic therapies. Recently, we demonstrated that the Polychaeta Eulalia viridis, common in the Portuguese rocky intertidal, holds a high variety of novel greenish and yellowish porphyrinoid pigments, stored as granules in the chromocytes of several organs. On the follow-up of this study, we chemically characterized pigment extracts from the worm's skin and proboscis using HPLC and evaluated their light and dark toxicity in vivo and ex vivo using Daphnia and mussel gill tissue as models, respectively. The findings showed that the skin and proboscis have distinct patterns of hydrophilic or even amphiphilic porphyrinoids, with some substances in common. The combination of the two bioassays demonstrated that the extracts from the skin exert higher dark toxicity, whereas those from the proboscis rapidly exert light toxicity, then becoming exhausted. One particular yellow pigment that is highly abundant in the proboscis shows highly promising properties as a natural photosensitizer, revealing that porphyrinoids from marine invertebrates are important sources of these high-prized bioproducts.

A Comparative Analysis of the Venom System between Two Morphotypes of the Sea Anemone Actinia equina.

The current study investigates the venom-delivery system of green and red morphotypes of the sea anemone Actinia equina to disclose its potential as a source of bioactive compounds. We compared the two morphotypes using electron and optical microscopy, proteomics, and toxicity assessment on zebrafish embryos. Specialized venom-injecting cells (nematocysts) are equally distributed and found in the tentacles of both varieties. Proteomics revealed proteins of interest in both red and green Actinia, yielding the three most abundant Gene Ontology (GO) terms related to the biological processes "proteolysis", "hemolysis in another organism" and "lipid catabolic process". Neurotoxins and cytolytic toxins similar to known cnidarian toxins like PsTX-60A and AvTX-60A, for instance, were identified in both types. Extracts from green and red anemones were toxic to zebrafish embryos, with green anemone venom appearing to be more potent. The findings highlight the presence of proteinaceous toxins in A. equina and the potential for different varieties to possess distinct bioactive compounds. Notably, pore-forming toxins are suggested for molecular probes and immunotoxins, making them valuable assets for potential biotechnological and biomedical purposes.

An Exploration of Novel Bioactives from the Venomous Marine Annelid Glycera alba.

The immense biodiversity of marine invertebrates makes them high-value targets for the prospecting of novel bioactives. The present study investigated proteinaceous toxins secreted by the skin and proboscis of Glycera alba (Annelida: Polychaeta), whose congenerics G. tridactyla and G. dibranchiata are known to be venomous. Proteomics and bioinformatics enabled the detection of bioactive proteins that hold potential for biotechnological applications, including toxins like glycerotoxins (GLTx), which can interfere with neuromuscular calcium channels and therefore have value for the development of painkillers, for instance. We also identified proteins involved in the biosynthesis of toxins. Other proteins of interest include venom and toxin-related bioactives like cysteine-rich venom proteins, many of which are known to interfere with the nervous system. Ex vivo toxicity assays with mussel gills exposed to fractionated protein extracts from the skin and proboscis revealed that fractions potentially containing higher-molecular-mass venom proteins can exert negative effects on invertebrate prey. Histopathology, DNA damage and caspase-3 activity suggest significant cytotoxic effects that can be coadjuvated by permeabilizing enzymes such as venom metalloproteinases M12B. Altogether, these encouraging findings show that venomous annelids are important sources of novel bioactives, albeit illustrating the challenges of surveying organisms whose genomes and metabolisms are poorly understood.

An investigation into the toxicity of tissue extracts from two distinct marine Polychaeta.

The present study investigated the potential toxicity of venomous secretions of two polychaetes, Hediste diversicolor and Glycera alba (Annelida: Phyllodocida). Toxic activity of putative toxins, measured on mussel gills through the Comet assay, revealed higher effects caused by extracts from H. diversicolor skin and G. alba specialised, jawed proboscis, when compared to control. The results suggest that H. diversicolor secretes toxins via skin for protection against predators, contrarily to G. alba, who secretes toxins for predation.

Interannual abundance changes of gelatinous carnivore zooplankton unveil climate-driven hydrographic variations in the Iberian Peninsula, Portugal.

The persistent massive blooms of gelatinous zooplankton recorded during recent decades may be indicative of marine ecosystem changes. In this study, we investigated the potential influence of the North Atlantic climate (NAO) variability on decadal abundance changes of gelatinous carnivore zooplankton in the Mondego estuary, Portugal, over the period 2003-2013. During the 11-year study, the community of gelatinous carnivores encompassed a larger diversity of hydromedusae than siphonophores; the former dominated by Obelia spp., Lizzia blondina, Clythia hemisphaerica, Liriope tetraphylla and Solmaris corona, while the latter dominated by Muggiaea atlantica. Gelatinous carnivore zooplankton displayed marked interannual variability and mounting species richness over the period examined. Their pattern of abundance shifted towards larger abundances ca. 2007 and significant phenological changes. The latter included a shift in the mean annual pattern (from unimodal to bimodal peak, prior and after 2007 respectively) and an earlier timing of the first annual peak concurrent with enhanced temperatures. These changes were concurrent with the climate-driven environmental variability mainly controlled by the NAO, which displayed larger variance after 2007 along with an enhanced upwelling activity. Structural equation modelling allowed depicting cascading effects derived from the NAO influence on regional climate and upwelling variability further shaping water temperature. Such cascading effect percolated the structure and dynamics of the community of gelatinous carnivore zooplankton in the Mondego estuary.

Changes in zooplankton communities along a mercury contamination gradient in a coastal lagoon (Ria de Aveiro, Portugal).

The main objective of this paper was to evaluate the impact of mercury on the zooplankton communities' structure and functioning and their bioaccumulation patterns along a contamination gradient in a temperate coastal lagoon. Our results demonstrated that total abundance was not negatively affected by Hg contamination, since the most contaminated areas presented the highest values, being the copepod Acartia tonsa the dominant species, which means that it is a very well adapted and tolerant species to mercury. Nevertheless, negative effects were observed in terms of species diversity, since the most contaminated areas presented the lowest values of species richness, evenness and heterogeneity. Moreover, the spatial mercury gradient was reflected on the bioaccumulation patterns of the zooplankton communities. This reinforces the idea that zooplankton can be considered as an important vehicle of mercury transfer through the food pelagic web since it constitutes a primordial food resource for several commercial fish species.