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Lectins are a protein family, present in almost all living organisms and involved in different biological pathways, such as immune responses. The Fucose Binding Lectin (FBL), constitute the latest lectin family identified and characterized in fishes. The FBL family is constituted by a large number of proteins exhibiting multiples of the F-type motif, either tandemly arrayed or in mosaic combinations with other domains. In an early step a FBL has been isolated and characterized from serum of the Antarctic fish Trematomus bernacchii by affinity chromatography on fucose-agarose column. A clear Bacterial agglutinating activity (BA) towards different bacteria strains (Escherichia coli, Kokuria rhizophyla and Bacillus subtilis) and Hemagglutinating activity (HA) toward rabbit erythrocytes was induced from the serum as well from the purified protein and thus confirm its involvement in host pathogen interactions. In SDS-PAGE analysis, the FBL exhibited an apparent molecular weight of 30 kDa. This data is confirmed from the sequence of the F lectin recognised on the T. bernacchii transcriptome. The sequence shows a similar and coherent structure with a supposed Mw 32.16 kDa and an isoelectropoint of 5.21. Furthermore, sequencing the N-terminus confirmed the identity of the sequence runned on SDS PAGE and blotted on PVDF membrane. The HA activity was analyzed at different temperatures and it was maintained also at the physiological living low temperatures of this fish habitat (close to 0 °C). Therefore, in order to identify trends linked to cold adaptation in Antarctic fish, we present our hypothesis on the conformational change determined by the aminoacidic substitutions respect the others fish fucolectins living in warmer water on the light of the general phylogenetic scenario including the new preliminary data on sharks FBL

Fish are equipped with a complex lectin repertoire that, like mammals, are involved almost all the immune reactions. Carbohydrate recognition and interactions mediated by lectins have been recognized involved in vertebrate innate immunity, not only for recognition of potential pathogens, but also acting in the agglutination, immobilization and other functional steps. Fish are equipped with a complex lectin repertoire that, like mammals, are involved almost all the immune reactions. Carbohydrate recognition and interactions mediated by lectins have been recognized involved in vertebrate innate immunity, not only for recognition of potential pathogens, but also acting in the agglutination, immobilization and other functional steps. In fish, C and F types lectins, galectins, rhamnose-binding lectin (RBL) and pentraxin have been identified in both in both cartilaginous and bony fish. In addition, selectins and other genes have been found in the currently available fish genomes. On the basis of our results on F-type (FBL) and RBL lectins we showed that: -lectin repertoires in fish are highly diversified and include not only representatives of the lectin families; described in mammals, but also members of lectin families described for the first time in fish species like the FBL and RBL; -these lectins have been identified in the eggs and embryos but also are present in the serum; -tissue-specific expression and localization of the diverse lectin repertoires and their molecular partners is consistent with their distinct biological roles in innate and adaptive immunity; -although some lectins may bind endogenous ligands, others bind sugars on the surface of potential pathogens; in addition to pathogen recognition and opsonization, some lectins display additional effector roles, such as regulation of immune functions.

Trace metal concentrations were measured in different tissues of Sabella spallanzanii, Styela plicata and Mytilus galloprovincialis collected in Termini Imerese harbor, located in Sicily (Italy), in order to evaluate the potential use of these species as bioindicators. The higher bioaccumulation factors (BAFs) were calculated in tube of Sabella spallanzanii, except for As which shows the higher BAF in branchial crown of the same specie. Regarding the other species analyzed, higher BAFs were found in the digestive gland of Mytilus galloprovincialis. An exception is represented by Pb that is significantly more concentrated in branchial basket and tunic of Styela plicata. The BAFs here calculated, show that all the species analyzed are able to accumulate a certain amount of metals as a consequence of filter feeding mechanisms, and thus assess the suitability of the species Styela plicata, Sabella spallanzanii and Mytilus galloprovincialis as indicators of water quality. Particularly, the tube of Sabella spallanzanii is an important compartment in metal retention and the more suitable for the evaluation of the contamination caused by trace elements. This article is protected by copyright. All rights reserved.

The study of cnidarians immunity, as model systems of metazoans, lead additional informations on the first steps of the immunity evolution. The functions of the genes and cellular pathways in higher vertebrates are conserved during the evolution of metazoans, as shown by the discovery of homologues in cnidarians. These basal metazoans in fact, are far from "simples" in the range of methods at their disposal to deal with potential prey but also invading microbes and pathogens. They can give informations about the invertebrates innate immune repertoire. We investigated the immunobiology starting from the inflammatory response in Anemonia sulcata (Cnidaria: Anthozoa) following injection of substances different in type and dimension, to understand the effector mechanisms involved in this process. We observed clear, strong and specific reactions especially after injection of bacteria and the alteration of the expression of enzymes (protease, phosphatase and esterase), showing a correlation between the appearance of the inflammatory reaction and the modification of enzymatic activities. From cnidarian phylum a large number of toxic compounds have been isolated. Tissues and mucus produced by cnidarians may have a role in immune defense and contain a variety of toxins as neurotoxins, cytolysins and antimicrobial peptides, which can have multifunctional role. The bioactive molecules were purified by acid extraction and HPLC purifications and characterized through biological assays, mass spectroscopy and peptide synthesis. Here, we show the cnidarian bioactive molecules as antimicrobial peptides and enzymes in order to draw applications in fields ranging from pharmacology to cultural heritage. Particularly, in the control of the microbic growth and especially in the tuning of biocleaning protocols, bioactive molecules with proteasic and esterasic activity have been used. These novel enzymes are active at temperature lower than 30°C, they need a reduced time of application and are safety for both operators and environment. Thus they could provide an important contribution to the development of sustainable innovative protocols.

In the last decades biotechnology research provides sustainable alternatives to traditional procedures for preventive preservation of cultural assets. Recently, bioactive molecules (BMs) isolated from marine invertebrate organisms have been isolated and tested for bioremoval of protein layers (BMP) or to controlling microbial colonization (BMA), acting at temperature lower than 30°C. The Protease or Antimicrobial activity was tested on ad hoc assembled specimens and on different historic-artistic manufacts. In bio-removing protocol BMP molecules were applied as gelled solutions, in order to guarantees a selective action, respectful of constitutive materials and manufact integrity. Peculiarity of Protease bioactive molecules is the temperature of action, lower than 30°C. Instead, BMAs molecules have been tested to control bacteria and fungi colonization in laboratory specimens. In our hypothesis these novel molecules provide an important contribution to the development of innovative protocols safe both for the environment and conservator health, representing a valid alternative to traditional methods according to the preventive conservation and "Minimal Intervention" concept in restoration procedures.

The evolution of multicellular organisms was marked by adaptation to protect against pathogens. The mechanisms for discriminating the ''self'' from ''non-self” have evolved into a long history of cellular and molecular strategies from damage repair to the co-evolution of host-pathogen interaction. The phylum of Cnidaria is one of the first branches in the tree of animal life to provide crucial insights on the evolution of immunity. Sea anemones (Anthozoa, Cnidaria) are benthic sessile species able to maintain the integrity of the tissues and allorecognition in colonial forms and to differentiate between symbionts and pathogenic intruders. We investigated the inflammatory response in sea anemone Anemonia sulcata following injection of various substances different in type and dimension, and observed clear, strong and specific reactions especially after injection of Escherichia coli and Vibrio alginolyticus. In particular we studied the effect of injectioninoculation of different sized suspensions (LPS, Phenyl Sepharose, cellulose, red blood cells, bacteria) into the pedal disk, of specimens in order to observe differential physical, biochemical and enzymatic reactions. We have shown for the first time in cnidarians a specific cellular and molecular response against pathogenic bacteria. The first phenomenon we observed was a body and foot swelling, followed by the formation of a yellowish rejection zone . A further study was also carried out by a histological analysis for the characterization of the area of rejection.This specific response suggested an extrusion of the material damaged by the animal, probably from the pores cinclides from which protrude the acontia in the pedal disk, a defense mechanism that has not yet been found in other cnidarians. Moreover, we analyzed enzymatic activity of protease, phosphatase and esterase, showing how the injection of different bacterial strains alters the expression of these enzymes, that catabolize prey tissues, suggesting a correlation between the appearance of the inflammatory reaction and the modification of enzymatic activities. Our study shows for the first time, a specific inflammatory-like response rejection and enzymatic responses following injection of bacteria in Cnidarian.

In questo studio è stato valutato l’utilizzo di molecole antimicrobiche estratte da organismi marini invertebrati (Anthozoa), al fine di limitare o inibire la crescita microbica su materiali impiegati per il restauro di manufatti storico-artistici. I ceppi batterici e fungini utilizzati nei saggi antimicrobici, sono stati isolati da porzioni di tele, stratificate con colla, impiegate nella foderatura di dipinti. Mediante un approccio integrato che comprende tecniche di microscopia e tecniche molecolari, sono state identificate colonie batteriche appartenenti ai generi Enterobacter e Micrococcus e fungine appartenenti ai generi Aspergillus e Penicillium. L’attività antimicrobica delle molecole (BMA1, BMA2, BMA3) è stata testata sui ceppi microbici identificati, definendo le corrispondenti Concentrazioni Minime Inibenti (MIC) e le Concentrazioni Minime Battericide/Fungicide (MBC/MFC). Inoltre, l’attività antimicrobica è stata testata su provini assemblati ad hoc in laboratorio, realizzati con colla pasta stratificata su due tipi di tela (lino, sintetica), simulando un intervento di foderatura. La presenza delle molecole BMA nella colla pasta ha contrastato la crescita microbica, inibendola completamente per il campione BMA1 e parzialmente in presenza di BMA2 e BMA3. L’utilizzo di queste molecole nel campo della conservazione dei beni culturali, costituisce un importante contributo allo sviluppo di tecnologie innovative, nel rispetto delle procedure del restauro conservativo.

The immune system that plays a major role in determining host fitness in the wild, i.e. under the constraints imposed by ecology and life history. Permanent conflict interactions with the environment are the natural situation for a living creature. To partially resolve this, the immune system evolved and is characterized by an enormous variety of mechanisms and effectors, including the AMPs, although their specificity is poor. In fact, AMPs are universal and extremely successful in dealing with a huge range of pathogens, including bacteria, fungi, protozoa and viruses Amps are oligopeptides composed of varying number of amino acids with a broad spectrum of targeted organisms ranging from viruses to parasites. Some of these compounds have been investigated with a view to possible therapeutic use as an alarming increase of resistance of microorganisms to classical antibiotics has been reported. In the present study, using primers designed from coding sequences from M. galloprovincialis and RACE, the cDNA sequence of the defensin was cloned and characterizated from the Red Sea mussel Brachidontes pharaonis, a Lessepsian invasive species, was first recorded in the Mediterranean seven years after the opening of the Suez Canal in 1869. In the past 30 years it has become abundant in midlittoral and infralittoral rocky habitats, especially along the rocky shores of the Eastern Mediterranean Brachidontes pharaonis has spread as far west as Sicily, probably by ship fouling and generated intensive population. Analysis of the sequence of 262 nucleotides revealed in the B. pharaonis defensine (DefBp) the presence of an ORF coding for 81 amino acids consisting of a signal peptide of 23 aa in the amino terminal portion followed by the mature peptide of 35 a.a. and a carboxy-terminal extension position. The full-length amino acid sequence and the results of the alignment with the sequences of M. galloprovincialis indicate an identity of 93% and a similarity of 96% with the antimicrobial peptide MGD1 and an identity of 78% and a similarity of 84% with the antimicrobial peptide of the same MGD2 Mediterranean mussel. In this species truncated sequences of defensin, including amino acid replacement to maintain 3D structure and increased positive net charge, possess antibacterial, antiprotozoan and antiviral capabilities. HIS analysis revealed BpDef mRNA expressed in circulating hemocytes with small intra-cytoplasmic granules (B, D) and with large granules Sequence identity and the common conserved sequence characteristics show that BpDef belongs to the  defensin family of AMPs with a typical domain structurally characterized by a helix and two sheets  . Two predicted bactericidal stretches were found in correspondence of the mature peptide and the Boman index value indicated the strong antibacterial feature of defensin of B. pharaonis.

In the last decades molecular biology has provided innovative approaches in order to set up specific protocols for the conservation and restoration of cultural assets. In this study, which falls within the so called field of Blue-biotechnology, new bio-reactive peptides isolated from marine invertebrate organisrns (Cnidaria and Molluscs) were tested aim to bio-cleaning (proteolytic- peptides) the surfaces or to control (antimicrobial-peptides) the colonization of historic-artistic manufacts by fungi or bacteria. Particularly, the proteolytic-peptides showed hydrolytic activity, specific for animal-glue, in a range of temperatures of 4-37°C; than acting without heating the surface, by a contro Iled procedure and avoiding damages to the constitutive materials, ali criteria required for conservation /restoration procedures. These hydrolases allow to carry out the removal of the protein layers in a controlled way, acting at the same temperature of the environment where the objects are restored (19-25.5 0C). lnstead, the antirnicrobial-peptides have been tested on a plethora of fungi and bacteria colonies and a specific activity, against Micrococcus spp., have been showed. We are testing the antimicrobial-peptides as a "biologica! biocide" in order to protect both organic and inorganic substrates. We hypothesize that the use of these molecules will give an important contribution to the development of innovative and efficient technologies concerning bio-cleaning and antimicrobial growth protocols, according to conservative restoration procedures.