FACIT collagen (1alpha-chain) is expressed by hemocytes and epidermis during the inflammatory response of the ascidian Ciona intestinalis

Based on previous cloning and sequencing study, real-time PCR and in situ hybridization assays of the inflamed body wall of LPS-injected Ciona intestinalis showed the enhanced gene expression of a collagen with FACIT structural features (Ci-type IX-Col 1a-chain). By using specific antibodies raised against an opportunely chosen Ci-type IX-Col synthetic peptide, the fibroblast property of hemocytes challenged in vitro with LPS (at 4 h) was displayed by flow cytometry, while immunocytochemistry identified hemocytes with large granules (morula cells) as collagen-producing cells. Hemocyte lysate supernatant analyzed in immunoblotting contained a 60 kDa band identifiable as 1a-chain-Ci-type IX-C…

Localizzazione di un recettore glucocorticoideo DLGR1 nei tessuti del teleosteo Dicentrarchus labrax

Bacillus thuringiensis treatment modulate the HSP70 expression in larva and adult brain of Rhynchophorus ferrugineus

To study the pathogen-host relationship, we used the model of the entomopathogenic bacterium Bacillus thuringiensis (Bt) and Rhynchophorus ferrugineus, a quarantine pest that attacks palm trees. In particular, we focused on the Bt stress-induced infections. We studied the effect of Bt on larval and adult growth, and on the expression of the heat shock proteins (HSPs), rapidly synthesized in the cell after exposure to stress including pathogens. Bt has negative effects on larval and adult growth, on total hemocytes counts and on the hemocyte type. HSP70 was evaluated in the supernatant of the brain lysate obtained from larvae and adults fed with sublethal doses of Bt. HSP70 expression was mo…

AUTOPHAGY AND APOPTOSIS MODULATION BY AQUEOUS EXTRACTS FROM LEAVES AND RHIZOMES OF Posidonia oceanica ON HEPG2 HEPATOCARCINOMA CELLS

Preliminary evidence for the cytotoxic molecule in hemocyte supernatant lysate from Ciona intestinalis unilocular hemocytes

In vitro effect of cadmium and copper on separated blood leukocytes of Dicentrarchus labrax.

The immunotoxic effects of heavy metals on blood leukocytes of sea bass (Dicentrarchus labrax) were examined. The cells, separated by a discontinuous Percoll-gradients, were exposed in vitro to various sublethal concentrations of cadmium and copper (10(-7) M, 10(-5) M, and 10(-3) M) and their immunotoxic effect was then evaluated by measuring neutral red uptake, MU assay, DNA fragmentation and Hsp70 gene expression. First of all, we demonstrated that the cells treated in vitro could incorporate Cd and Cu. A relationship between heavy metal exposure and dose-time-dependent alterations in responses of leukocytes from blood was found for both metals, but copper was more immunotoxic than cadmiu…

Anthropic impact, bioactive molecules, sustainable development, my keyword? Biodiversity!

Biodiversity is my main research focus and that of my research group. We study a range of different aspects concerning biodiversity, such as the impact anthropic activities have on biodiversity and how to improve its preservation, its use as a valuable resource for the extraction of bioactive molecules, and the study of its distribution. The research group has over twenty years of experience in the study of zoology, and, more in particular, the immune and behavioural responses of aquatic invertebrates and fish. For many years, the group has observed the effects of various anthropogenic activities, such as the acoustic and chemical impacts on the physiological and behavioral responses of inv…

Cytotoxic activity of Holothuria tubulosa (Echinodermata) coelomocytes.

Abstract The immune system of marine invertebrates, in particular that of holothurians, still requires further study. Our research showed that coelomocyte cells contained in the coelomic fluid of the sea cucumber, Holothuria tubulosa, are able to lyse, in vitro, red blood cells in rabbits and sheep. A plaque-forming assay showed spherule cells to be the effector cells, able to release cytotoxic molecules after xenogenic cell contact. The coelomocyte lysate supernatant, analysed by polyacrylamide gel electrophoresis overlay technique, using rabbit and sheep erythrocytes, showed two different haemolytic protein patterns: one calcium dependent and the other calcium independent. The fractions o…

Spontaneous cytotoxic activity of eosinophilic granule cells separated from the normal peritoneal cavity ofDicentrarchus labrax

Abstract In this study the spontaneous in vitro cytotoxic activity to tumour cell lines, (K562), by unstimulated sea bass ( Dicentrarchus labrax ) leukocytes was examined by trypan blue exclusion test and lactate dehydrogenase release assay. A high anti-tumour cell line activity of resident peritoneal leukocytes was found at an effector to target ratio (E:T) of 25:1 after incubation for 2 h at 18° C. Rabbit and sheep erythrocytes were not lysed. A low activity was displayed by head kidney and spleen cell populations whereas blood leukocytes revealed no significant activity. The effect of E:T ratio on cytotoxicity as well as microscopy observations suggested that the cytotoxic reaction requi…

Galectine nella risposta infiammatoria di Ciona intestinalis cDNA ed espressione genica

Expression and distribution of DIGR1 in the ontogeny of Dicentrarchus labrax

Fatty Acids and Triacylglycerols Profiles from Sicilian (Cold Pressed vs. Soxhlet) Grape Seed Oils

Among the Sicilian economic productive sectors, that of wine production has today a considerable economic value. However, with the growth of this sector, notable was the increase in the production of waste, which to date is not only an economic damage for companies, but also a threat to the environment. It is known that waste from wine production has properties (e.g., antioxidants) which have potential reuse at cosmetic, pharmaceutical and nutritional levels to obtain economically sustainable applications. A new goal is given by the recovery of added value compounds from agri-food wastes and by-products. Grape seed oil is a promising vegetable fat and cold pressing does not involve the use …

Prophenoloxidase activating system in tunicate hemolymph

The activation sequence and related factors of the prophenoloxidase activating system in crustaceans was compared with the equivalent system in tunicates. Both solitary and colonial ascidians present in their hemolymph a copper-dependent phenoloxidase activity that may be inhibited by tropolone and phenylthiourea. Carbohydrates are able, to various extents, to trigger proPO system which requires serine protease cleavage for activation to phenoloxidase (PO). In some ascidians, hemocytes called ≪morula cells≫ show PO activity, while in Ciona intestinalis the ≪univacuolar refractile granulocytes≫ are positive after cytochemical staining with L-dopa. The relationships between proPO system and d…

EFFECT OF EXTRACTS FROM LEAVES AND RHIZOMES OF THE SEAGRASS POSIDONIA OCEANICA ON HEPG2 HEPATOCARCINOMA (HCC) CELLS

Modulazione In vivo dell’Hsp70 negli emociti di Rhynchophorus ferrugineus dopo trattamento con Bacillus thuringiensis

Diversi tipi di stress ambientali possono indurre l’espressione delle Hsps vengono rapidamente sintetizzate all'interno delle cellule dopo esposizione ad uno stressor. Le HSPs sono raggruppate in diverse famiglie in base alle dimensioni delle proteine. Nella maggior parte degli organismi ci sono diversi geni che codificano i membri di questa famiglia HSP. In studi precedenti è stata valutata l'interazione tra Bacillus thuringiensis e gli emociti circolanti di Rhynchophorus ferrugineus il punteruolo rosso delle palme. Questo coleottero è grande, da due a cinque centimetri di lunghezza, ed è di colore rosso ruggine, le sue larve scavano buche fino a un metro lungo il tronco delle palme, e può…

Cytotoxic Potential of the Coelomic Fluid Extracted from the Sea Cucumber Holothuria tubulosa against Triple-Negative MDA-MB231 Breast Cancer Cells

Growing evidence has demonstrated that the extracts of different holothurian species exert beneficial effects on human health. Triple negative breast cancers (TNBC) are highly malignant tumors that present a poor prognosis due to the lack of effective targeted therapies. In the attempt to identify novel compounds that might counteract TNBC cell growth, we studied the effect of the exposure of the TNBC cell line MDA-MB231 to total and filtered aqueous extracts of the coelomic fluid obtained from the sea cucumber Holoturia tubulosa, a widespread species in the Mediterranean Sea. In particular, we examined cell viability and proliferative behaviour, cell cycle distribution, apoptosis, autophag…

Survey on the presence of A. simplex s.s. and A. pegreffii hybrid forms in Central-Western Mediterranean Sea

Abstract In this study, 1029 fish and cephalopod samples came from Central-Western Mediterranean (FAO 37.1.1 and FAO 37.1.3) were analysed for Anisakidae larvae research with the aim to identify possible hybridisations between Anisakis pegreffii and Anisakis simplex s.s. species. A total of 1765 larvae were detected, with prevalence values between 8.1% and 100%. The morphologic analysis revealed characters attributable to morphotype I of Anisakis in 98.5% of the examined larvae, while 1.5% belonged to the morphotype II. PCR-based Restriction Fragment Length Polymorphism (PCR–RFLP) analysis of the entire ITS region (ITS1, 5.8S and ITS2) of nuclear ribosomal DNA (rDNA) was performed with Hinf…

Seabream (Sparus aurata) hierarchy among alfa and beta subordinates and dominant interplay affects stress responses and phagocytic activity by peritoneal cavity cells

Toxic mineral elements in Mytilus galloprovincialis from Sicilian coasts (Southern Italy)

We assessed the relationship between V, Cr, Mn, Hg, As, Cd, Sn, Sb and Pb concentrations in Mytilus galloprovincialis samples from the coasts of Sicily and the expression of metallothioneins. Toxic mineral elements assessment was carried out by A.A. Spectrometry and ICP-MS. The metallothioneins expression was performed by q-PCR method. Low metals' levels were found in the mussel samples examined, in comparison with what was reported in literature. The highest mean values of toxic mineral elements were found in Gela (Cr 0.178 +/- 0.03 mg/Kg, Mn 4.325 +/- 0.012 mg/Kg, As 3.706 +/- 0.009 mg/Kg, Sn 0.148 +/- 0.014 mg/Kg, Sb 0.009 +/- 0.004 mg/Kg e Pb 0.364 +/- 0.01 mg/Kg). Significant levels of…

Putative rhamnose-binding lectin in the solitary ascidian Ciona intestinalis.

Ciona robusta macrophage migration inhibitory factor (Mif1 and Mif2) genes are differentially regulated in the lipopolysaccharide-challenged pharynx

The effects of lipopolysaccharide (LPS) on Mif (macrophage migration inhibitory factor) gene expression in the pharynx (haemapoetic tissue) of Ciona robusta were investigated using quantitative reverse-transcription PCR (qRT-PCR) and in situ hybridisation (ISH). To verify the induction of an inflammatory response in the pharynx, a qRT-PCR analysis was performed to evaluate the change in the expression of proinflammatory marker genes such as Mbl, Ptx-like, Tnf-α and Nf-kb, which were shown to be upregulated 1 h post LPS challenge. The change in the expression of the two Mif paralogs in the pharynx was assessed before and after stimulation, and qRTPCR and ISH results showed that, although Mif…

Attività antibatterica di peptidi estratti da celomociti di echinodermi

Negli echinodermi la difesa dell’organismo dalle invasioni microbiche avviene tramite meccanismi cellulari e umorali dell’immunità innata. Una componente fondamentale dei fattori umorali è costituita dai peptidi antimicrobici (AMPs) molto conservati con un ampio spettro di attività antimicrobica contro batteri, virus e funghi. Solitamente gli AMPs sono costituiti da meno di 100 amminoacidi, sono cationici e formano strutture anfipatiche. In questo lavoro dimostriamo che i celomociti dell’Echinoideo Paracentrotus lividus e dell’Oloturoideo Holothuria tubulosa contengono sostanze con attività antimicrobica verso diversi ceppi batterici patogeni umani. In entrambe le specie, nel precipitato ac…

Effects of Social Hierarchy Establishment on Stress Response and Cell Phagocytosis in Gilt-Head Sea Bream (Sparus aurata)

Social stress can affect the ability of fish to respond to various stressors, such as pathogens or environmental variations. In this paper, the effects of social stress on gilt-head bream (Sparus aurata) were investigated. To study the effects of physiological stress, we evaluated biochemical and cellular parameters, such as cortisol, glucose, lactate, osmolarity, and phagocytosis, 24 h after the establishment of social hierarchy in a group of three fish. Social hierarchy was determined and characterized by behavioral observation (aggressive acts and feeding order) of the specimens (dominant: “α”; subordinate: “β” and “γ”). After the esta…

Components of hemocyte extracts from marine invertebrates exert antimicrobial activity

Cellule che producono ciTNFalfa sono attive nell'infiammazione e nello sviluppo larvale di Ciona intestinalis

A D-galactose specific lectin is an inducible inflammatory IL-1-like opsonin in the hemolymph of the ascidian Ciona intestinalis challegend with LPS.

Attivazione del sistema della profenolossidasi nella risposta infiammatoria della tunica di Ciona intestinalis (L.)

Citocentrifugación como un método adicional para estudiar celomitos de equinodermos: un enfoque comparativo que combina células vivas, preparaciones teñidas y espectroscopía de rayos-x de dispersión de energía

Abstract Introduction: Echinoderm coelomocytes have traditionally been investigated through a morphological approach using light microscopy, which relies on the idea of constant cell shape as a stable character. However, this can be affected by biotic or abiotic conditions. Objective: To analyze if the consistency in cell morphology offered by the cytocentrifugation method, might be used as a convenient tool to study echinoderm coelomocytes. Methods: Cells of Echinaster (Othilia) brasiliensis (Asteroidea), Holothuria (Holothuria) tubulosa (Holothuroidea), Eucidaris tribuloides, Arbacia lixula, Lytechinus variegatus, and Echinometra lucunter (Echinoidea) were spread on microscope slides by c…

Cell-Free Coelomic Fluid Extracts of the Sea Urchin Arbacia lixula Impair Mitochondrial Potential and Cell Cycle Distribution and Stimulate Reactive Oxygen Species Production and Autophagic Activity in Triple-Negative MDA-MB231 Breast Cancer Cells

Triple-negative breast cancer (TNBC) is a highly malignant tumor histotype which lacks effective targeted therapies, thereby being considered as the most aggressive form of breast carcinoma. To identify novel compounds which could counteract TNBC cell growth, we explored the in vitro effects of crude extracts and &lt

CHERAX DESTRUCTOR (CLARK, 1836) AND CHERAX QUADRICARINATUS (VON MARTENS, 1868): SAFETY AND NUTRITIONAL QUALITY- Premio "YOUNG INVESTIGATOR AWARD"

Two species of freshwater Parastacidae (Cherax quadricarinatus and Cherax destructor) are used for breeding in several country and to date also in Italy in one aquaculture plant located near Catania (Sicily). Although they are Australian, they seem to have some of the peculiar properties that make it important, strategic and highly appreciated species for aquaculture facilities in Italy. In fact, they mature early, have multiple reproductive cycles in a year, females can lay over a thousand eggs in a single brood and seem to tolerate wide ranges of temperature (Haubrock et al., 2021). Several authors have studied the Cherax genus from different points of view: diseases, moulting phases and …

Bright spots in the darkness of cancer: A review of starfishes-derived compounds and their anti-tumor action

The fight against cancer represents a great challenge for researchers and, for this reason, the search for new promising drugs to improve cancer treatments has become inevitable. Oceans, due to their wide diversity of marine species and environmental conditions have proven to be precious sources of potential natural drugs with active properties. As an example, in this context several studies performed on sponges, tunicates, mollusks, and soft corals have brought evidence of the interesting biological activities of the molecules derived from these species. Also, echinoderms constitute an important phylum, whose members produce a huge number of compounds with diverse biological activities. In…

Novel antimicrobial and anti-biofilm agents from a marine invertebrate

seabream(Sparus aurata)hierarchies of social behaviour affects stress responses and immunity

Low frequencies noise effects on behaviour of Sparus aurata juveniles

Human activities in the oceans, such as marine traffic and Deep Sea Mining (DSM), are increasing in the last years (Ross 2005; Calvo et al 2016). Underwater noise in the oceans, especially at lower frequencies, is mainly produced by marine traffic and DSM could increase this noise in the next future. Marine Strategy Framework Directive promotes the achievement of a good quality environmental status and aims to monitor “continuous low frequency sound” trend in the ambient noise level within the 1/3 octave bands centred at 63 and 125 Hz. The aim of this study is to evaluate the behavioural changes of Sparus aurata juveniles exposed to a four different acoustic signals in a tank. The emitted s…

AQUATIC ACOUSTIC NOISE: BEHAVIORAL AND MOLECULAR RESPONSES IN ECHINODERMS, THE CASE OF A. LIXULA (LINNAEUS, 1758) SEA URCHINS

Anthropic noise is considered a real pollutant, in particular the submarine noise. The impact on biodiversity is not yet sufficiently understood. Further research is needed to evaluate any negative effects. The noises associated with anthropogenic activities are increasing: shipments, seismic surveys, sonar, recreational rowing and future mineral extraction activities from ocean depths (DSM). These noises are having an impact on the welfare of many marine species. The understanding of the effects on biodiversity could be important for the creation of guidelines, laws or rules for the most environmentally sustainable exploitation of natural resources. Our study aims to investigate the motili…

Lectine sieriche di "tipo F" nell'immunità innata dei pesci: aspetti molecolari e funzionali.

Purification and characterization of D-galactose binding lectin involved in the inflammatory response in Ciona intestinalis

Effects of exogenous cortisol on the expression of glucocorticoid receptor (DIGR1) and hsp70 in head-kidney and peritonel cavity cells from Dicentrarchus labrax.

Further insight on Ciona intestinalis prophenoloxidase system activated during the LPS induced inflammatory response

Effetti del cadmio sull'espressione delle HSP70 nelle cellule del sangue di Spigola (Dicentrachus labrax L. Osteichthyes, Moronidae)

A lytic mechanism based on soluble phospholypases A2 (sPLA2) and b-galactoside specific lectins is exerted by Ciona intestinalis (ascidian) unilocular refractile hemocytes against K562 cell line and mammalian erythrocytes

Abstract Hemocytes from the ascidian Ciona intestinalis exert in vitro Ca 2+ -dependent cytotoxic activity toward mammalian erythrocytes and K562 cells. To examine the lytic mechanism, hemocyte populations were separated (B1–B6 bands) through a Percoll discontinuous density gradient, the hemocyte cytotoxic activity (HCA) and the lytic activity of the hemocyte lysate supernatant (HLS) were assayed. In addition the separated hemocytes were cultured and the cell-free culture medium (CFM) assayed after 3 h culture. Results support that unilocular refractile hemocytes (URGs), enriched in B5, are cytotoxic. The B5-HLS contains lysins and the activity of B5-CFM shows that lysins can be released in…

A glucocorticoid receptor in sea bass leukocytes

Seabream (Sparus aurata) long-term dominant-subordinate interplay affects phagocytosis by peritoneal cavity cells.

Fish are sensitive to stressful conditions that affect their innate immune systems and increase their susceptibility to diseases. We examined the social stress of paired gilthead seabream (Sparus aurata). Social hierarchies (dominant/subordinate) were characterised by behavioural changes, such as “aggressiveness” and “feeding order”; hierarchical positions were established within an hour of exposure to social stress and remained unchanged for approximately 1 year. To characterise physiological stress, we measured blood plasma levels of cortisol, glucose, and lactate as well as osmolarity and observed that the levels of these stress markers were higher in subordinate individuals than in domi…

Aroclor 1254 inhibits the chemiluminescence response of peritoneal cavity cells from sharpsnout sea bream (Diplodus puntazzo)

Chronic exposure to polychlorinated biphenyls (PCBs) affect the immune system of fish and could lead to a decreased disease resistance. The effects of Aroclor 1254, PCB mixtures, on the Diplodus puntazzo innate immunity were examined by assaying the zymosan stimulated chemiluminescence response (CL) of peritoneal cavity cells (PCCs) at various times (1, 24, 48 h and 1-4 weeks) from intraperitoneal injection of the xenobiotic (1 mg kg(-1) body weight). Controls were performed by assaying cells from medium-treated fish. Since the kinetic of the chemiluminescence response showed the highest peak at 25 min after the zymosan stimulation of the cells, the values found at that time were considered…

Chitosan Film Functionalized with Grape Seed Oil—Preliminary Evaluation of Antimicrobial Activity

Although the fishing and wine industries undoubtedly contribute significantly to the economy, they also generate large waste streams with considerable repercussions on both economic and environmental levels. Scientific literature has shown products can be extracted from these streams which have properties of interest to the cosmetics, pharmaceutical and food industries. Antimicrobial activity is undoubtedly among the most interesting of these properties, and particularly useful in the production of food packaging to increase the shelf life of food products. In this study, film for food packaging was produced for the first time using chitosan extracted from the exoskeletons of red shrimp (Ar…

Serum lectins in fish innate immunity: molecular and functional aspects

Fucose-binding lectins (FBL) are present in tissues and fluids from invertebrates and vertebrates. The lectin repertoires in teleost fish are highly diversified and recently has been described the structure of the fucose-binding agglutinin that revealed a novel lectin fold (the “F-type” eel (Anguilla anguilla) fold), which shared a unique fucose-binding sequence motif contained both in carbohydrate-binding proteins and unrelated proteins. In this report, we describe serum FBL from sea bass Dicentrarchus labrax and sea bream Sparus aurata. These lectins were purified, characterized, cloned and sequenced. Studies on structural aspects, biological activity, tissue distribution as well as ontog…

Noise elicits hematological stress parameters in Mediterranean damselfish (Chromis chromis, perciformes): A mesocosm study

In the last few decades, technological developments and the widespread rise of anthropic activities have increased the exposure of organisms to noise pollution, thus evoking great interest in its biological effects, particularly on the immune system. The aim of the present work was to investigate some of the biochemical parameters in the blood of Chromis chromis (Linnaeus, 1758) following in vivo exposure to noise levels of 200 and 300 Hz. Our results revealed that, compared to the control specimens, the fish exposed to noise had significantly increased levels of stress biomarkers such as glucose, lactate and total proteins in plasma, as well as a rise in the expression of heat shock protei…

Immunological Activity of Ascidian Hemocytes

In ascidians, various hemocyte types and their differentiation stages may be responsible for several immune functions. A central role in the immune effector mechanisms can be assigned to PO-containing hemocytes and prophenoloxidase system. Morula cell in Styela plicata and univacuolar refringent granulocyte in Ciona intestinalis, a probable intermediate stages in the differentiation pathway of morula cell, may be cytotoxic cells. They are involved in immune reactions of solitary and colonial ascidians. We have shown that they are provided with a PO-linked spontaneous cytotoxic activity and recognize mammalian erythrocytes or tumor cells markers. The toxic molecules can be radical oxygen int…

Behavioural and biochemical stress responses of Palinurus elephas after exposure to boat noise pollution in tank

This study examined the effects of boat noise on the behavioural and biochemical parameters of the Mediterranean spiny lobster (Palinurus elephas).The experiment was conducted in a tank equipped with a video and audio recording system. 18 experimental trials, assigned to boat noise and control conditions, were performed using lobsters in single and group of 4 specimens. After a 1. h habituation period, we audio- and video-recorded the lobsters for 1. h. During the experimental phase, the animals assigned to the boat groups were exposed to boat noise pollution (a random sequence of boat noises). Exposure to the noise produced significant variations in locomotor behaviours and haemolymphatic …

Effetti dell’esposizione a cadmio e rame sull’attività di cellule del sangue di spigola (Dicentrarchus labrax L. Osteichthyes, Moronidae).

Cadmium, Copper and Tributyltin effects on fertilization of Paracentrotus lividus (Echinodermata)

Marine environments are continuously being threatened by a large number of xenobi- otics from anthropogenic sources. The effect of chemical pollution on living organisms are numerous and may impair reproductive success of adults species of marine invertebrate and vertebrate through effects on gamete quality. Echinoderms are characterized by external fertilizzation and gametes, free of any type of protection, may be in contact with toxic substances so the reproductive success depends largely on the environment conditions. The purpose of this work is to assess the effects on the in vitro fertiliza- tion of exposure of sea urchin Paracentrotus lividus gametes to xenobiotic substances as CuSO4,…

In vivo modulation of Hsp70 in Rhynchophorus ferrugineus hemocytes after Bacillus thuringiensis treatment

Heat shock proteins (Hsps) are rapidly synthesized within stressed cells after exposure to an environmental stressor. A variety of environmental stresses, including heat, cold, trace-metal exposure, xenobiotics have been reported to modulate Hsps expression in various organisms. Hsps are grouped into several families based on their protein size. Most organisms have several genes encoding members of this Hsp family. In particularly Hsp70 can be induced quickly under stressful conditions, but return to a normal expression level under non-stressful conditions. Few studies have been done to detect the Hsp70 expression in phytophagous insects towards pathogens. Since a preliminary research discl…

Hemocyte types and some plasmatic properties of two edible crabs Cancer borealis and Cancer pagurus

Cancer pagurus and Cancer borealis and are edible crabs produced by economically relevant aquaculture. In this study the hemocytes and some plasmatic parameters of Cancer borealis and Cancer pagurus were examined. The cell features of the hemocytes were observed using light and scanning electron microscopy (SEM). Granulocytes, semigranulocytes and hyalinocytes were mainly identified on the basis of size, presence/absence and quantity of the cytoplasmic granules and the nucleus-to-citoplasma (N/C) ratio. SEM observations were useful for disclosing the surface features of these cells, and the same characteristics were found in both crab species. A smooth surface distinguishes elongated hyalin…

Biological responses to high-frequency sound in a freshwater crayfish, Cherax quadricarinatus

Anthropogenic noise is recognised as an important environmental stressor that can have longterm negative consequences on species. In recent years, there has been increasing attention to the potential negative impact of noise pollution on species, with great concern for the importance of impacts on aquatic animal life. This study examined the effects of acoustic stress on the biochemical parameters of the freshwater crayfish Cherax quadricarinatus. The experiment was conducted in a tank equipped with an audio and video recording system using ten groups (five control and five test) of three adult crayfish (30 animals in total). The animals in the test group were exposed to acoustic signals [a…

The prophenoloxidase system is activated during the tunic inflammatory reaction of Ciona intestinalis

Phenoloxidase (PO) activity was examined in the tunic tissue of Ciona intestinalis following lipopolysaccharide (LPS) intratunic injection. Tunic homogenate supernatant (THS), assayed with the Dopa-MBTH reaction, displayed Ca(2+)-independent PO activity that was raised by LPS and further enhanced by proteases. Specific inhibitors (tropolone, phenylthiourea, diethylthiocarbamate) supported the specificity of the reaction. Assay with soybean trypsin inhibitor showed that, in the tunic, PO activation with trypsin was not significantly inhibited suggesting that proteases diverse from serine proteases were involved. In vivo experiments were carried out by injecting isosmotic medium or LPS, and T…

EFFECTS OF ACUSTIC STRESS ON BIOCHEMICAL AND MOBILITY PARAMETERS AND BEHAVIOUR IN THE CRAYFISH, CHERAX DESTRUCTOR

Invertebrate lectins present cytokine properties.

Inflamed adult pharynx tissues and swimming larva of Ciona intestinalis share CiTNFalfa-producing cells

Invertebrate lectins present cytockine properties.

Expression and distribution of the glucocorticoid receptor DlGR1 in the teleost Dicentrarchus labrax brain

Cortisol is the main corticosteroid secreted by the interrenal cells of the head kidney and it exerts a role in mantaining the omeostatic status in fish. In teleosts its effects are mediated through intracellular receptors expressed in several tissues, that are ligand-dependent transcription factors by binding to specific tissue DNA sequences. In Dicentrarchus labrax we previously cloned and sequenced a glucocorticoid receptor, DlGR1, isolated from leukocytes of peritoneal cavity. In this work we showed mRNA expression and tissue immunohistochemical localization of brain DlGR1 by in situ hybridization assays, with a riboprobe with DlGR1 cDNA trascriptional activation domain, and by immunohi…

Biological activity, tissue distribution and preliminary moleular characterization of a serum fucolectin from the sea bass (Dicentrarchus labrax)

Peritoneal cavity phagocytes from the teleost sea bass express a glucocorticoid receptor (cloned and sequenced) involved in genomic modulation of the in vitro chemiluminescence response to zymosan

Abstract To gain further insight into the role of cortisol in Wsh innate immune responses, we cloned and sequenced a 2592 bp cDNA from sea bass (Dicentrarchus labrax) peritoneal leukocytes (PCLs) encoding a glucocorticoid receptor (DlGR1). The deduced aminoacid sequence displayed that DlGR1 belong to a multigenic family of steroid hormone receptors, and exhibited high homology (80%) to the Burton’s mouth breeder (Haplochromis burtoni) HbGR1. The DlGR1 functional domains presented homologies with those of several vertebrate species. In situ hybridization assay revealed that DlGR1 was expressed in macrophages and neutrophils from the peritoneal cavity. Since in a previous paper, sea bass PCL …

Effects of waterg gun on echinoderms: biochemical changes on sea urchin (Arbacia lixula) and sea cucumber (Holothuria tubulosa)

Sources Airguns and Waterguns are used in seismic exploration the marine seafloor for different purposes such as oil and gas search, or geological-geodynamic reconstruction. In this technique a bubble of compressed air is produced with a consequently production of loud sound wave penetrating the ocean floor. The impact of seismic Airguns was assessed in some cetaceans (Kyhn et al. 2019), fish species and invertebrate (Carroll et al. 2017), but we need to know more. In this study we evaluate the biochemical changes in sea urchin (Arbacia lixula) and sea cucumber (Holothuria tubulosa) exposed to a 20 minutes of watergun noise (1 pulse each 10 s) at a distance of 50 m. For each species 40 indi…

Attività citotossica dei celomociti di Holothuria tubulosa (Echinodermata)

Numerosi studi hanno accertato che nel fluido celomatico degli echinodermi sono presenti cellule con attività diretta verso agenti estranei. In particolare i celomociti svolgono un repertorio di funzioni immunitarie tra cui la fagocitosi, la citotossicità, l’attività antibatterica, la formazione di capsule e il rigetto dei trapianti. Nel presente lavoro abbiamo studiato l’attività citotossica dei celomociti di Holothuria tubulosa (Oloturoideo) non separati ed abbiamo utilizzato cellule di mammifero come target. I celomociti prelevati dal fluido celomatico lisano gli eritrociti di coniglio o di montone e le cellule tumorali della linea eritromieloide umana K562. Inoltre anche il supernatante…

Vessel noise pollution as a human threat to fish: assessment of the stress response in gilthead sea bream (Sparus aurata, Linnaeus 1758)

This study examined the effects of boat noise pollution on the stress indices of gilthead sea bream (Sparus aurata, Linnaeus 1758). To assess the stress response in these fish, biometric values and plasma parameters such as ACTH, cortisol, glucose, lactate, haematocrit, Hsp70, total protein, cholesterol, triglycerides and osmolarity were analysed. After acclimatization of the animals, the experiment was carried out in a tank fitted with underwater speakers where the fish were exposed to sound treatments (in duplicate) consisting of: 10 days of no sound (control treatment; the animals were only exposed to the experimental tank’s background noise) and 10 days of noise derived from original re…

Enhanced expression of a cloned and sequenced Ciona intestinalis TNFa-like (CiTNFa) gene during the LPS-induced inflammatory response.

A tumor necrosis factor-alpha (TNFalpha)-like gene from Ciona intestinalis (CiTNF alpha-like) body wall challenged with bacterial lipopolysaccharide (LPS) was cloned and sequenced 4 h after LPS inoculation. An open reading frame of 936 bp encoding a propeptide of 312 amino acids (35.4 kDa) displaying a transmembrane domain from positions 7 to 29, a TACE cleavage site, and a mature peptide domain of 185 amino acids (20.9 kDa), was determined with a predicted isoelectric point of 9.4. The phylogenetic tree based on deduced amino acid sequences of invertebrate TNF-like protein and vertebrate TNFs supported the divergence between the ascidian and vertebrate TNF families, whereas D. melanogaster…

FISH WASTE, NOT A PROBLEM BUT A POSSIBLE SOLUTION: CHITOSAN EXTRACTS AND NEW PACKAGING POSSIBILITIES.

The fishing industry is one of the oldest industries in Europe and also one of the most important economic resources in the Mediterranean area. However, to date, in addition to creating considerable economic wealth, the various sectors (markets, restaurants, shops, etc.) also create significant levels of fish waste. If not processed or recycled adequately, these waste streams can lead to environmental degradation and severe pollution (Gao et al., 2018). Although today fish waste is considered a potential resource to generate high-added value, current practices in the fishing industry do not fully exploit its biological potential. Amongst the various possibilities for reuse, the production o…

Nuovi dati sulla risposta infiammatoria dell’ascidia Ciona intestinalis

Human Drug Pollution in the Aquatic System: The Biochemical Responses of Danio rerio Adults

Simple Summary The release of medicinal products for human use in the aquatic environment is now a serious problem, and can be fatal for the organisms that live there. Danio rerio is a freshwater fish that provides the possibility to study the effects of these pollutants on the health of aquatic organisms. The results of the various existing scientific studies are scarce and conflicting. Here, we review the scientific studies that have analyzed these effects, highlighting that the impacts of drugs are evident in the biochemical responses of these animals. Abstract To date, drug pollution in aquatic systems is an urgent issue, and Danio rerio is a model organism to study the toxicological ef…

New Bioactive Peptides from the Mediterranean Seagrass Posidonia oceanica (L.) Delile and Their Impact on Antimicrobial Activity and Apoptosis of Human Cancer Cells

The demand for new molecules to counter bacterial resistance to antibiotics and tumor cell resistance is increasingly pressing. The Mediterranean seagrass Posidonia oceanica is considered a promising source of new bioactive molecules. Polypeptide-enriched fractions of rhizomes and green leaves of the seagrass were tested against Gram-positive (e.g., Staphylococcus aureus, Enterococcus faecalis) and Gram-negative bacteria (e.g., Pseudomonas aeruginosa, Escherichia coli), as well as towards the yeast Candida albicans. The aforementioned extracts showed indicative MIC values, ranging from 1.61 μg/mL to 7.5 μg/mL, against the selected pathogens. Peptide fractions were further analyzed thr…

Glucocorticoid receptor (DlGR1) is expressed in pre-larval and larval stages of the teleost fish Dicentrarchus labrax.

Glucocorticoid hormone receptors (GR), members of the nuclear hormone receptor superfamily, are ligand-dependent transcription factors expressed in various tissues by binding to specific DNA sequences. Since glucocorticoids have a role in maintaining the homeostatic status in fish, we previously cloned and sequenced a GR (DlGR1) of adult Dicentrarchus labrax; we also showed mRNA expression (in situ hybridization) and tissue immunohistochemical localization of DlGR1 in several organs. This work has now been extended to the examination of the expression, tissue distribution, and cytolocalization of DlGR1 in larval developmental stages by similar methods to those used for the adult organs. The…

Tunicate immunocytes can be cytotoxic toward foreign cells

Effects of Sulfamethoxazole on Fertilization and Embryo Development in the Arbacia lixula Sea Urchin

To date, drugs released into the aquatic environment are a real problem, and among antibiotics, sulfamethoxazole is the one most widely found in wastewater; thus, the evaluation of its toxicity on marine organisms is very important. This study, for the first time, investigates the in vitro effects of 4 concentrations of sulfamethoxazole (0.05 mg/L, 0.5 mg/L, 5 mg/L, 50 mg/L) on the fertilization and development of the sea urchin Arbacia lixula. The gametes were exposed to drugs in three different stages: simultaneously with, prior to, and post-fertilization. The results show a significant reduction in the percentage of fertilized oocytes at the highest drug concentrations. Moreover, an incr…

Sphingomyelin as well as carbohydrates are involved in the mechanism of cytotoxic molecules contained and released in vitro by Ciona intestinalis granulocytes.

Espressione e distribuzione del recettore DLGR1 nel cervello del Teleosteo Dicentrarchus labrax.

Cellular responses and HSP70 expression during wound healing in Holothuria tubulosa (Gmelin, 1788).

Wound repair is a key event in the regeneration mechanisms of echinoderms. We studied, at the behavioural, cellular and molecular levels, the wound healing processes in Holothuria tubulosa after injuries to the body wall. The experiments were performed for periods of up to 72 h, and various coelomocyte counts, as well as the expression of heat shock proteins (HS27, HSP70 and HSP90), were recorded. Dermal wound healing was nearly complete within 72 h. In the early stages, we observed the injured animals twisting their bodies to keep their injuries on the surface of the water for the extrusion of the buccal pedicles. At the cellular level, we found time-dependent variations in the circulating…

Mercury in fish products: what’s the best for consumers between bluefin tuna and yellowfin tuna?

A total of 205 bluefin and yellowfin tuna samples were examined for mercury detection in order to verify possible differences and have a detailed risk assessment of the two tuna species. The results showed significant higher mercury concentration in muscle tissue of bluefin tuna respect yellowfin tuna (p < 0.001) with mean concentration of 0.84 mg/kg and maximum value of 1.94 mg/kg. These differences can be due the different biological and ecological aspects of the two tuna species and to different oceanographic aspects between Atlantic Ocean and Mediterranean sea. The results obtained in this study suggest an advisable containment of the sources of pollution and further studies on the c…

Espressione di recettori glucocorticoidei in cellule dell’immunità innata di Dicentrarchus labrax

HPLC/HRMS and GC/MS for Triacylglycerols Characterization of Tuna Fish Oils Obtained from Green Extraction

Background: Fish oil is one of the most common lipidic substances that is consumed as a dietary supplement. The high omega-3 fatty acid content in fish oil is responsible for its numerous health benefits. Fish species such as mackerel, herring, tuna, and salmon are particularly rich in these lipids, which contain two essential omega-3 fatty acids, known as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Objectives: Due to the scarcity of information in the literature, this study aimed to conduct a qualitative and quantitative characterization of triglycerides (TAGs) in crude tuna fish oil using HPLC/HRMS. Fatty acid (FA) determination was also performed using GC/MS. The tuna fis…

Are mussels able to distinguish underwater sounds? Assessment of the reactions of Mytilus galloprovincialis after exposure to lab-generated acoustic signals

This study examined the effects of lab-generated acoustic signals on the behaviour and biochemistry of Mediterranean mussels (Mytilus galloprovincialis). The experiment was carried out in a tank equipped with a video-recording system using six groups of five mussels exposed to five acoustic treatments (each treatment was replicated three times) for 30 min. The acoustic signals, with a maximum sound pressure level of 150 dB rms re 1 μPa, differed in frequency range as follows: low (0.1–5 kHz), mid-low (5–10 kHz), mid (10–20 kHz), mid-high (20–40 kHz) and high (40–60 kHz). The exposure to sweeps did not produce any significant changes in the mussels' behaviour. Conversely, the specimens expos…

Espressione e distribuzione del recettore DLGR1 nell’ontogenesi di Dicentrarchus labrax.

Polyphenol Characterization and Antioxidant Activity of Grape Seeds and Skins from Sicily: A Preliminary Study

The interest in the consumption of health-promoting foods has led to identifying derivatives of the wine industry as products to increase the functional properties of different foods or to design new functional foods. The main goal of this study is to characterize and valorize byproducts and wastes of Sicilian grapes as new sources of bioactive components, from the perspective of a circular economy and a biorefinery approach. In particular, this research investigated: 1. the total phenolic content and antioxidant activities and 2. the phenolic profiles of free and bound fractions of defatted grape seeds and red grape skins from Sicily. Defatted grape seeds (DGS) and red grape skins (RGSK) a…

Profiling the physiological and molecular response to sulfonamidic drag in Procambarus clarkii

Sulfamethoxazole (SMZ) is one of the most widely employed sulfonamides. Because of the widespread use of SMZ, a considerable amount is indeed expected to be introduced into the environment. The cytotoxicity of SMZ relies mainly on arylhydroxylamine metabolites (S-NOH) of SMZ and it is associated with the production of reactive oxygen species (ROS). There is limited information about the toxic potential of SMZ at the cellular and molecular levels, especially in aquatic and/or non-target organisms. In the present study, the red swamp crayfish (Procambarus clarkii), being tolerant to extreme environmental conditions and resistant to disease, was used as a model organism to profile the molecula…

Parametri sierologici e cellulari della specie alloctona Procambarus clarckii

Procambarus clarkii è un animale d’acqua dolce originario del Centro-Sud degli Stati Uniti e del Nord-Est del Messico. Questa specie rientra nella lista delle 100 specie invasive più pericolose al mondo (Delivering Alien Invasive Species Inventories for Europe) in quanto presenta una serie di caratteristiche biologiche, che la rendono una minaccia per la biodiversità. In Sicilia la sua presenza è stata registrata per la prima volta nel 2003 nella riserva naturale “Lago Preola e Gorghi Tondi”, Mazara del Vallo (TP). Allo scopo di conoscere meglio la biologia di questa specie, su esemplari di P. clarkii provenienti dalla riserva di Trapani, abbiamo esaminato i tipi emocitari ed alcune attivit…

Modulation of prophenoloxidase activity in Ciona intestinalis inflammatory response

Anaphylatoxin-like molecules generated during complement activation induce a dramatic enhancement of particle uptake in rainbow trout phagocytes.

Here we have identified a serum fraction containing approximately 8-kDa molecules with an unexpected capacity to greatly enhance particle uptake in trout head kidney leukocytes (HKLs). This approximately 8-kDa particle-uptake enhancing fraction (PUEF-8) was purified from complement-activated serum by gel filtration chromatography. Mass spectrometric analysis and reactivity of anti-trout C3-1 and C4 antibodies, indicated the presence of C3a, C4a and C5a molecules in PUEF-8. Using a newly developed flow cytometric assay that measures the capacity of cells to ingest fluorescent beads, we showed that PUEF-8 induced a striking enhancement (344+/-50% higher than the PBS control value) in the numb…

The immune mediators in echinoderms as souce of novel AMPs against microbial biofilms

A Novel Peptide with Antifungal Activity from Red Swamp Crayfish Procambarus clarkii

The defense system of freshwater crayfish Procambarus clarkii as a diversified source of bioactive molecules with antimicrobial properties was studied. Antimicrobial activity of two polypeptide-enriched extracts obtained from hemocytes and hemolymph of P. clarkii were assessed against Gram positive (Staphylococcus aureus, Enterococcus faecalis) and Gram negative (Pseudomonas aeruginosa, Escherichia coli) bacteria and toward the yeast Candida albicans. The two peptide fractions showed interesting MIC values (ranging from 11 to 700 μg/mL) against all tested pathogens. Polypeptide-enriched extracts were further investigated using a high-resolution mass spectrometry and database search and 14 n…

Differential expression of two glucocorticoid receptors in seabass (teleost fish) head kidney after exogeneous cortisol inoculation

Stressful conditions include a prompt release of corticosteroid hormones which can mediate gene expression through glucocorticoid receptors (GR). Since two seabass (Dicentrarchus labrax) GRs have been cloned and sequenced from peritoneal cavity cells (DlGR1) and liver (DlGR2), a comparative amino acid sequence analysis that included Haplochromis burtoni HbGRs, was carried out and homologies disclosed. The DlGR1 and DlGR2 deduced aminoacid sequences showed 61% identity (I) and 70% similarity (S). Moreover, DlGR2 was similar to HbGR2b (69% I, 73% S), and the DlGR1 to HbGR1 (72% I, 78% S). In addition, we examined the expression of the DlGRs after exogeneous cortisol inoculation into the perit…

Antitumoral compounds from vertebrate sister group: A review of Mediterranean ascidians

Among the diseases that afflict the human population, cancer is one for which many drug treatments are not yet known or effective. Moreover, the pharmacological treatments used often create serious side effects in sick patients and for this reason, it is essential to find effective and less harmful treatments. To date, marine biodiversity is a real source of metabolites with antitumoral activity and among invertebrates' ascidians have been the main source to obtain them. Mediterranean area is the richest in biodiversity and contains several ascidian species used in drugs development during the years. However, many more Mediterranean ascidian species have not been studied and could be a sour…

Vitamin D3 affects innate immune status of European sea bass (Dicentrarchus labrax L.)

The effects of vitamin D3 dietary administration on certain innate immune parameters on the expression of immune-related genes in head-kidney (HK) and gut were investigated in European sea bass Dicentrarchus labrax. Vitamin D3 (vD3) was orally administered to fish in a commercial pellet food supplemented with 0 (control); 3750; 18,750; or 37,500 U kg−1. Furthermore, gut histology was considered. This study showed a modulation in the activities examined in fish fed with the addition of vD3. After just 2 weeks of administration, diet supplementation with the vitamin resulted in increased phagocytic ability, while serum peroxidase content was increased in fish fed with all experimental diets a…

Survey on the presence of non-dioxine-like PCBs (NDL-PCBs) in loggerhead turtles (Caretta caretta) stranded in south Mediterranean coasts (Sicily, Southern Italy).

A total of 71 loggerhead turtles (Caretta caretta) stranded along the coasts of Sicily (Southern Italy) were examined for non–dioxine like polychlorinated biphenyl (NDL-PCB) levels in muscle and adipose tissue by a gas chromatography-mass spectrometry/mass spectrometry (GC-MS/MS) method. The results revealed 6 high-indicator congener (∑6PCBIND) levels in 45% of the loggerhead turtles examined, with mean values of 980.39 ± 2508.39 ng/g wet weight in adipose tissue and 102.53 ± 238.58 ng/g wet weight in muscle tissue. The hexachloro and heptachloro PCB congeners were the most abundant in both the sample types. The highest NDL-PCB levels were reached in an adipose tissue sample of a logg…

The cytotoxic activity of Ciona intestinalis (ascidian) unilocular refractile hemocytes versus K562 tumor cells and mammalian erythrocytes involves phospholypase A2 and lectins

F-type lectin from the sea bass (Dicentrarchus labrax): purification, cDNA cloning, tissue expression and localization, and opsonic activity.

Recently described biochemical and structural aspects of fucose-binding lectins from the European eel (Anguilla anguilla) and striped bass (Morone saxatilis) led to the identification of a novel lectin family ("F-type" lectins) characterized by a unique sequence motif and a characteristic structural fold. The F-type fold is shared not only with other members of this lectin family, but also with apparently unrelated proteins ranging from prokaryotes to vertebrates. Here we describe the purification, biochemical and molecular properties, and the opsonic activity of an F-type lectin (DlFBL) isolated from sea bass (Dicentrarchus labrax) serum. DlFBL exhibits two tandemly arranged carbohydrate-r…

Effect of in vitro exposure to cadmiumand copper on sea bass blood cells

Blood cells freshly collected from sea bass (Dicentrarchus labrax) were exposed in vitro to different concentrations of cadmium (Cd) and copper (Cu) at 10-7 M, 10-5 M, 10-3 M, and exam- ined for neutral red retention capacity and for cell vitality with MTT assay. A relationship between heavy metal exposure and alteration in responses of blood cells in a dose-time-dependent was found. Our results showed that fish blood cells may constitute an interesting biological model for experimen- tal and applied toxicology, especially in the case of environmental pollution.

An alternative set of test to bioassay for bioinsecticides

The development of an assay to determine insecticidal properties for either biological and conventional plant protection products plays an important role on the early screening of potential pathogens or derived toxins candidates. The standard methods for the evaluation it has been by bioassay, especially determination of LD50 or LC 50 requiring the use of relatively large numbers of insects and toxin tests. There are several problems connected with these bioassays: availability of insects and in the right life stage, mass producing the candidate species, preparation, reproducibly and costs relative to intensive manpower. These aspects are really important especially when bio-insectides shou…

Inflamed adult pharynx tissues and swimming larva of Ciona intestinalis share CiTNFalpha-producing cells.

In situ hybridisation and immunohistochemistry analyses have shown that the Ciona intestinalis tumour necrosis factor alpha gene (CiTNFalpha), which has been previously cloned and sequenced, is expressed either during the inflammatory pharynx response to lipopolysaccharide (LPS) or during the swimming larval phase of development. Granulocytes with large granules and compartment/morula cells are CiTNFalpha-producing cells in both inflamed pharynx and larvae. Pharynx vessel endothelium also takes part in the inflammatory response. Haemocyte nodules in the vessel lumen or associated with the endothelium suggest the involvement of CiTNFalpha in recruiting lymphocyte-like cells and promoting the…

ANTIMICROBIAL ACTIVITY FROM POLYPEPTIDE-RICH EXTRACTS OF THE SEAGRASS POSIDONIA OCEANICA

The present study was carried out to assess the antibacterial, antifungal and antibiofilm properties of polypeptide-rich extracts isolated from green leaves and rhizomes of Mediterranean seagrass Posidonia oceanica (L. Delile) (Posidoniaceae). The seagrass was collected, washed with freshwater, grinded with liquid nitrogen in order to obtain fine powders that were exposed to extraction by acetic acid and antiproteases. The crude extracts isolated from leaves and rhizomes of P. oceanica were subjected to microbiological assays to evaluate the antibacterial, antifungal and antibiofilm activity of polypeptide fraction against two reference bacterial strains Staphylococcus aureus ATCC 25923 and…

Localizzazione di una lectina di “tipo F” nell’adulto e nell’ontogenesi di Dicentrarchus labrax

The purification, cloning, sequencing, molecular properties and expression of a fucose-binding lectin from the serum of Dicentrarchus labrax (DlFBL) have been previously reported. We now describe the distribution and expression of DlFBL during fish ontogeny. Immunohistochemistry and in situ hybridization assays were carried out at various developmental stages (from 10 days posthatching larvae to juveniles). Another fucose-binding lectin, similar to DlFBL in biochemical, immunochemical and agglutinating properties, was extracted and purified from eggs and appeared to be localized in the embryo yolk sack residual. DlFBL was found in columnar and goblet cells of the intestinal epithelium of la…

The effect of low frequency noise on the behaviour of juvenile Sparus aurata.

[EN] Anthropogenic activities are causing increased noise levels in the marine environment. To date, few studies have been undertaken to investigate the effects of different noise frequencies on the behaviour of juvenile fish. In this study, the behavioural changes of juvenile gilthead seabream (Sparus aurata) are evaluated when exposed to white noise filtered in third-octave bands centred at 63, 125, 500, and 1000 Hz (sound pressure level, 140-150 dB re 1 mu& x3a1;a) for 7 h. The group dispersion, motility, and swimming height of the fish were analysed before and during the acoustic emission. Dispersion of the fish was found to reduce immediately upon application of low frequency sound (63…

Underwater high frequency noise: Biological responses in sea urchin Arbacia lixula (Linnaeus, 1758)

Marine life is extremely sensitive to the effects of environmental noise due to its reliance on underwater sounds for basic life functions, such as searching for food and mating. However, the effects on invertebrate species are not yet fully understood. The aim of this study was to determine the biochemical responses of Arbacia lixula exposed to high-frequency noise. Protein concentration, enzyme activity (esterase, phosphatase and peroxidase) and cytotoxicity in coelomic fluid were compared in individuals exposed for three hours to consecutive linear sweeps of 100 to 200 kHz lasting 1 s, and control specimens. Sound pressure levels ranged between 145 and 160 dB re 1μPa. Coelomic fluid was …

Humoral responses during wound healing in Holothuria tubulosa (Gmelin, 1788)

Abstract Wounds in living organisms trigger tissue-repair mechanisms. The sea cucumber (Holoturia tubulosa) is an excellent model species for achieving a better understanding of the humoral and cellular aspects involved in such healing processes. Consequently, this study assesses data on its morphometric, physiological and humoral responses 1, 2, 6, 24 and 48h after wound induction. In particular, morphometric data on the weight, width, length and coelomic-fluid volume of the species were estimated at different times during our experiments. In addition, the humoral aspects related to the enzymatic activity of esterase, alkaline phosphatase and peroxidase, as well as the cytotoxic activity o…

Stress e immunità nei gamberi allevati

Integration of pomace and grape seeds in feed of broiler chickens: effect on the chemical characteristics of the meat

Recently, consumer demand for healthier, safer, good quality food products has increased. The use of grape pomace and grape seeds, natural antioxidants rich in polyphenols and known for their ability to prevent lipid oxidation, has attracted significant interest and could become an important alternative as a partial substitute for vitamin E which is the most commonly used antioxidant in animal diets. In this context, the valorization and reuse of wine industry wastes could be a way to reduce costs for companies and damage to the environment [1]. The objective of this study was to evaluate the effect of adding different percentages of pomace and grape seeds to the diet of broiler chickens. T…

CHEMICAL CHARACTERIZATION AND BIOLOGICAL PROPERTIES OF MACROALGAE EXTRACTS

Marine species are a rich source of bioactive molecules and among them, it is known that marine algae produce different secondary metabolites for which different biological activities such as: immunomodulatory (Raposo et al., 2016), antioxidant (Fisch et al., 2003) and antimicrobial (Pinteus et al., 2015) were demostrated. The aim of this study was chemical charac- terize the extracts of three macroalgae species: Carpodesmia crinite (Duby, Orellana & Sansón, 2019), Carpodesmia brachy- carpa (J. Agardh, Orellana & Sansón 2019, WoRMS, 2023), Ericaria brachycarpa (J. Agardh, Molinari & Guiry, 2020), Asparagopsis taxiformis (Delile) Trevisan 1845) to evaluate also their biological a…

Chemiluminescence response of ß-glucan stimulated leukocytes isolated from different tissues and peritoneal cavity of Dicentrarchus labrax

Abstract The respiratory burst of leukocytes isolated from sea bass ( Dicentrarchus labrax ) pronephros, peritoneal cavity (P.C.), spleen and blood, was measured by a chemiluminescence (CL) assay after stimulation with β-glucan. The CL response by P.C. and pronephros leukocytes was significantly higher than that expressed by a similar number of cells separated from spleen and blood. This probably reflects the observation that the proportion of macrophages and neutrophils was highest in the populations of leukocytes from peritoneal cavity and pronephros. Comparative observations showed a higher degree of yeast phagocytosis by leukocytes taken from peritoneal cavity than the pronephros. Moreo…

Expression of a glucocorticoid receptor (D1GR1) in several tissues of the teleost fish Dicentrarchus labrax

Since glucocorticoids have a role in maintaining the homeostatic status in fish, in the present paper mRNA expression (in situ hybridization) and tissue immunohistochemical localization of a glucocorticoid receptor (DlGR1) in several Dicentrarchus labrax organs are reported. Riboprobe and specific antibodies were prepared by using the DlGR1 that has been previously cloned and sequenced from peritoneal cavity leukocytes. Both mRNA and receptor were identified in head kidney, spleen, gills, intestine, heart and liver tissues. The functional roles of DlGR1 localization are discussed.

Biological and Proteomic Characterization of the Anti-Cancer Potency of Aqueous Extracts from Cell-Free Coelomic Fluid of Arbacia lixula Sea Urchin in an In Vitro Model of Human Hepatocellular Carcinoma

Echinoderms are an acknowledged source of bioactive compounds exerting various beneficial effects on human health. Here, we examined the potential in vitro anti-hepatocarcinoma effects of aqueous extracts of the cell-free coelomic fluid obtained from the sea urchin Arbacia lixula using the HepG2 cell line as a model system. This was accomplished by employing a combination of colorimetric, microscopic and flow cytometric assays to determine cell viability, cell cycle distribution, the possible onset of apoptosis, the accumulation rate of acidic vesicular organelles, mitochondrial polarization, cell redox state and cell locomotory ability. The obtained data show that exposed HepG2 cells under…

Histone Deacetylase Inhibitors from Marine Invertebrates

Simple Summary Histone deacetylases (HDACs) are enzymes that control gene expression and are involved in the onset of serious human pathologies, including cancer; hence, their inhibitors (HDACis) have received increased attention in recent years. It is known that marine invertebrates produce significant amounts of molecules showing active pharmacological properties and an extensive spectrum of biomedical applications. This review is focused on the description of the molecular, biochemical, and, where available, physiological aspects of marine invertebrate-derived compounds that possess HDACi properties, taking into consideration their possible utilization as treatment agents against differe…

Use of a comprehensive diagnostic algorithm for Anisakis allergy in a high seroprevalence Mediterranean setting

Summary Background.Diagnosis of anisakis allergy (AA) is based on the skin prick test (SPT) and specific IgE (sIgE) determination. Anyway, false positivity cases are due to cross reactivity with numerous allergens. The aim of the study was to evaluate the reliability of a comprehensive diagnostic algorithm for the AA. Methods.An observational study was conducted on a sample of consecutive subjects accessing the allergology outpatient ambulatories of two hospitals located in Western Sicily. All the recruited outpatients were tested by Skin Prick Test performed using anisakis extracts by ALK-Abello (Madrid, Spain). Specific IgE dosage for anisakis extracts was then performed by using ImmunoCA…

Survey on the presence of non-dioxine-like PCBs (NDL-PCBs) in loggerhead turtles (Caretta caretta) stranded in south Mediterranean coasts (Sicily, Southern Italy)

A total of 71 loggerhead turtles (Caretta caretta) stranded along the coasts of Sicily (Southern Italy) were examined for non-dioxine like polychlorinated biphenyl (NDL-PCB) levels in muscle and adipose tissue by a gas chromatography-mass spectrometry/mass spectrometry (GC-MS/MS) method. The results revealed 6 high-indicator congener (∑6 PCBIND ) levels in 45% of the loggerhead turtles examined, with mean values of 980.39 ± 2508.39 ng/g wet weight in adipose tissue and 102.53 ± 238.58 ng/g wet weight in muscle tissue. The hexachloro and heptachloro PCB congeners were the most abundant in both the sample types. The highest NDL-PCB levels were reached in an adipose tissue sample of a loggerhe…

The potential of antimicrobial peptides isolated from freshwater crayfish species in new drug development: A review

Abstract The much-publicised increased resistance of pathogenic bacteria to conventional antibiotics has focused research effort on the characterization of new antimicrobial drugs. In this context, antimicrobial peptides (AMPs) extracted from animals are considered a promising alternative to conventional antibiotics. In recent years, freshwater crayfish species have emerged as an important source of bioactive compounds. In fact, these invertebrates rely on an innate immune system based on cellular responses and on the production of important effectors in the haemolymph, such as AMPs, which are produced and stored in granules in haemocytes and released after stimulation. These effectors are …

EFFECT OF SUBMARINE ACOUSTIC NOISE IN JUVENILE SEA BREAM (SPARUS AURATA) AND MUSSELS (MYTILUS GALLOPROVINCIALIS)

The earth’ resources are running out, the population will increase and further sources will be needed. These, were found in the deep ocean. To date the mining activities (Deep Sea Mining, DSM) they have not started. One impact of DSM could be underwater noise. Considering the noise frequencies of anthropic activities in the ocean, and the European directive on underwater noise control, we stressed the animals with four acoustic 1/3 band noises around: 63 Hz, 125 Hz, 500 Hz, 1kHz. We study the effects on Sparus aurata juveniles, and on Mytilus galloprovincialis. We will analyse behavioural., molecular and biochemical responses Los recursos de la tierra se están agotando, la población aumenta…

Expression of a type IX-like collagen in tissue injury of the ascidian Ciona intestinalis durino inflammatory process

Espressione e distribuzione del recettore DLGR1 nel cervello del teleosteo Dicentrarchus labrax (L. 1758)

Risposta allo stress ambientale di Dentex dentex, Pagrus major ed il loro ibrido “Pantice”.

Effetti modulanti del cadmio e del rame sulle attività cellulari del teleosteo Dicentrarchus labrax

Elevated cortisol modulates hsp70 and hsp90 gene expression and protein in sea bass head kidney and isolated leukocytes

In fish, interactions between Hsps and cortisol are involved in stress modulated physiological processes including innate immune responses. Cortisol exerts a role in the regulation of Hsps synthesis. Fish head kidney is a lymphomieloid and endocrine organ releasing cortisol, and it is the central organ for immune-endocrine interactions. In sea bass, cortisol intraperitoneal injection and in vitro treatment of head kidney cells show that inducible Hsp70 and Hsp90 are modulated by this hormone. However, an inverse relationship between mRNA expression (real-time PCR) and Hsp70 and Hsp90 protein levels (densitometric band analysis) was found. Time-course assays indicate a cortisol-mediated regu…

Does the immune answer to Bacillus thuringiensis infection is the same in larvae, females and males of Rhynchophorus ferrugineus?

Bacillus thuringiensis (Bt) è considerato un potenziale batterio entomopatogeno per la lotta al Rhynchophorus ferrugineus, coleottero da quarantena infestante le palme. In questo lavoro si considerano gli effetti dell’infezione di Bt in larve, maschi e femmine del punteruolo rosso. La patogenicità è stata valutata stimando la LD50 (Lethal Median Dose) e la LT50 (Lethal Median Time), il numero totale di emociti ed il tipo di emociti (Differential Haemocytes counts) ed inoltre l’espressione delle proteine Heat Shock Protein 70 (HSP70) negli emociti e nel cerebro. La mortalità di entrambi i sessi e delle larve aumentava all’aumentare della dose e del tempo di esposizione. Tuttavia le larve han…

Studio preliminare sulla caratterizzazione degli emociti di Cancer pagurus

Il sistema della profenolossidasi nella risposta infiammatoria della tunica di Ciona intestinalis (L.)

Does Bacillus thuringiensis Affect the Stress and Immune Responses of Rhynchophorus ferrugineus Larvae, Females, and Males in the Same Way?

Simple Summary Rhynchophorus ferrugineus is a destructive quarantine pest of palm trees, now widely distributed. Although broad-spectrum insecticides are often used to protect palm against R. ferrugineus, there is increasing concern about their effects on the environment and human health, especially where palm trees are located in urban areas. As an environmentally friendly entomopathogen, Bacillus thuringiensis (Bt) has been widely used to prevent other pest infestations. Although Bt products are the most sold bio-insecticides, there are still many interesting features to be investigated in the relationship of Bt and its hosts. We investigated the effect of Bt on larvae, females, and males…

Bacillus thuringiensis treatment alters larval growth, hemocytes and modulation of Hsp70 in Rhynchophorus ferrugineus

To study the pathogen-host relationship, we used the model of the entomopathogenic bacterium Bacillus thuringiensis (Bt) and Rhynchophorus ferrugineus, a quarantine pest that attacks the palm trees. In particular, we focused on the Bt stress-induced infections. We studied the effect of Bt on larval growth, on hemocytes and on the expression of the heat shock proteins (Hsp70). HSPs are rapidly synthesized in the cells after a stress exposition including pathogens. The Hsp70 was evaluated in the supernatant of the hemocyte lysate (HLS) obtained from larvae fed with Bt. This is the first time that the presence of Hsp70 has been recorded in R. ferrugineus. Bt has negative effects on larval grow…

Salmo salar fish waste oil: Fatty acids composition and antibacterial activity

Background and aims Fish by-products are generally used to produce fishmeal or fertilizers, with fish oil as a by-product. Despite their importance, fish wastes are still poorly explored and characterized and more studies are needed to reveal their potentiality. The goal of the present study was to qualitatively characterize and investigate the antimicrobial effects of the fish oil extracted from Salmo salar waste samples and to evaluate the potential use of these compounds for treating pathogen infections. Methods Salmo salar waste samples were divided in two groups: heads and soft tissues. Fatty acids composition, and in particular the content in saturated (SAFAs), mono-unsaturated (MUFA…

In vitro effects of methylmercury on ascidian (Styela plicata) immunocyte responses

This study shows that high methylmercury concentrations are cytotoxic for Styela plicata hemocytes, whereas sublethal concentrations affect immunocyte responses. Moreover, hemocytes exposed to the xenobiotic present a significantly enhanced phenoloxidase activity as revealed in the hemocyte lysate supernatant compared with the control. Although the cytotoxic activity of S. plicata hemocytes toward rabbit erythrocytes is a PO-dependent cell-target reaction due to quinone products, it was significantly decreased by suitable methylmercury concentrations in the medium. The same xenobiotic concentrations decreased the hemocyte phagocytic activity toward yeast. In both the responses cell-target c…

ANTIMICROBIAL AND ANTIBIOFILM ACTIVITY OF A PEPTIDE IDENTIFIED IN THE SEAGRASS POSIDONIA OCEANICA

Attività biologica, distribuzione tissutale e caratterizzazione molecolare della fucolectina sierica di spigola (Dicentrarchus labrax).

Shipping noise affecting immune responses of European spiny lobster (Palinuruselephas)

Marine crustaceans are influenced by numerous environmental factors. Environmental stress from pollutants seems to affect their metabolism, growth, moulting, survival, and immune defence. Recently, it has become clear that there is an impact of the global increase in sea noise levels due to shipping traffic on crustacean welfare. Considering the ecological and commercial importance of the European spiny lobster (Palinurus elephas (Fabricius, 1787)) in most parts of the Mediterranean coastal area, in the present study we investigated whether shipping noise pollution contribute to changing the immune parameters of stress in the European spiny lobster. The animals were exposed to a mix of nois…

In vivo modulation of Bacillus thuringiensis (commercial form) on Holothuria tubulosa immune defense mechanisms

In invertebrates, cellular and humoral components are evolved to maintain their homeostais and integrity. Both these components respond to different non-self such as microorganisms, vertebrate erythrocytes and foreign proteins. The aim of this research was to study the possible modulating effects of Bacillus thuringiensis (Bt) in a commercial and vegetative form on the immune response of the echinoderm, sea cucumber Holothuria tubulosa. Bt is one of the most used biopesticides worldwide, covering the 90% of the organic market. Sicily is a region vocated to biological control and agricultural area are often closed to the coastal area. Investigate the effect of biopesticides on other non targ…

A serum fucolectin isolated and characterized from sea bass Dicentrarchus labrax

A lectin specific for fucose and galactose was isolated by affinity chromatography on Sepharose CL-6B from the serum of Dicentrarchus labrax. The hemagglutinating activity against rabbit erythrocytes was calcium-independent, and reached its maximum at 37 degrees C. Two protein components were found in the hemagglutinating fractions eluted from the Sepharose column. Only the 34 kDa component (DLL2) eluted from the polyacrylamide gels (SDS-PAGE) showed agglutinating activity against rabbit erythrocytes. SDS-PAGE, in non-reducing conditions, revealed a single 66 kDa protein that reacted with antibodies to the 34 kDa component. Therefore, a dimeric structure stabilized by disulfide bonds can be…

In Vitro Cytotoxic Effect of Aqueous Extracts from Leaves and Rhizomes of the Seagrass Posidonia oceanica (L.) Delile on HepG2 Liver Cancer Cells: Focus on Autophagy and Apoptosis

Aqueous extracts from Posidonia oceanica’s green and brown (beached) leaves and rhizomes were prepared, submitted to phenolic compound and proteomic analysis, and examined for their potential cytotoxic effect on HepG2 liver cancer cells in culture. The chosen endpoints related to survival and death were cell viability and locomotory behavior, cell-cycle analysis, apoptosis and autophagy, mitochondrial membrane polarization, and cell redox state. Here, we show that 24 h exposure to both green-leaf- and rhizome-derived extracts decreased tumor cell number in a dose–response manner, with a mean half maximal inhibitory concentration (IC50) estimated at 83 and 11.5 μg of dry extract/mL, respecti…

Stress and immune response to bacterial LPS in the sea urchin Paracentrotus lividus (Lamarck, 1816).

The immune system of the sea urchin species Paracentrotus lividus is highly complex and, as yet, poorly understood. P. lividus coelomocytes mediate immune response through phagocytosis and encapsulation of non-self particles, in addition to the production of antimicrobial molecules. Despite this understanding, details of exactly how these processes occur and the mechanisms which drive them are still in need of clarification. In this study, we show how the bacterial lipopolysaccharides (LPS) is able to induce a stress response which increases the levels of the heat shock proteins HSP70 and HSP90 only a few hours after treatment. This study also shows that LPS treatment increases the expressi…

An Attempt to re-examine the immune role of Ciona intestinalis hemocytes

Aspetti filogenetici della nuova famiglia di lectine "di tipo F" nei Pesci.

Evoluzione dell’immunità innata. Componenti della reazione infiammatoria indotta da LPS nella parete corporea dell’ascidia Ciona intestinalis.

The Role of Spatial Exploration and Territoriality in Establishing Gilthead Seabream (Sparus aurata) Hierarchies, and Their Effects upon Underlying Stress Physiology

Territoriality, spatial exploration and social hierarchy are strictly related behaviors in gregarious fishes, and are often under-appreciated in farms where the individuals are confined within crowded spaces. In this study, we investigated the role of spatial exploration, elucidating the importance of time upon forming the social organization, and the role of the territoriality in gilthead seabream (Sparus aurata), using two experimental approaches. In the first approach, three fish were placed sequentially in the aquarium with an interval of two days (sequential model), while in the second (simultaneous model), two fish were simultaneously placed in an aquarium divided by a barrier which w…

Gender differences in the immune system activities of sea urchin Paracentrotus lividus

In the immune system of vertebrates, gender-specific differences in individual immune competence are well known. In general, females possess more powerful immune response than males. In invertebrates, the situation is much less clear. For this purpose we have chosen to study the immune response of the two sexes of the echinoderm Paracentrotus lividus in pre- and post-spawning phases. The coelomic fluid from the echinoderms contains several coelomocyte types and molecules involved in innate immune defenses. In this article we report that the degree of immune responses in the P. lividus differs according to sex in both pre- and post-spawning phases. We found in all tests that females were mor…

Anthropogenic noise: the behavioural responses of Sparus aurata juveniles as the basis for a numerical model.

Underwater noise is mainly produced by anthropogenic activities and has increased in recent years. The Marine Strategy Framework Directive aims to monitor “continuous low frequency sound” trends in ambient noise in particular at 1/3rd octave bands centered at 63 and 125 Hz. The aim of this work is to evaluate the possibility of using the results of Sparus aurata juvenile’s behavior for parameterizing a numerical model of the fish behavior. The fishes were stressed with white noise filtered with a band pass filter at 1/3 octave band centered respectively at 63 Hz, 125 Hz, 500 Hz, and 1 kHz (SPL: 140-150 dB re 1µPa). We performed three test replicas for the control (without any sound emission…

I pesci "taggati"

Cytotoxic capability and the associated proteomic profile of cell-free coelomic fluid extracts from the edible sea cucumber Holothuria tubulosa on HepG2 liver cancer cells

Hepatocellular carcinoma (HCC) is an aggressive cancer histotype and one of the most common types of cancer worldwide. The identification of compounds that might intervene to restrain neoplastic cell growth appears imperative due to its elevated overall mortality. The marine environment represents a reservoir rich in bioactive compounds in terms of primary and secondary metabolites produced by aquatic animals, mainly invertebrates. In the present study, we determined whether the water-soluble cell-free extract of the coelomic fluid (CFE) of the edible sea cucumber Holothuria tubulosa could play an anti-HCC role in vitro by analyzing the viability and locomotory behavior, cell cycle distribu…

Immune mediators of sea-cucumber Holothuria tubulosa (Echinodermata) as source of novel antimicrobial and anti-staphylococcal biofilm agents

The present study aims to investigate coelomocytes, immune mediators cells in the echinoderm Holothuria tubulosa, as an unusual source of antimicrobial and antibiofilm agents. The activity of the 5kDa peptide fraction of the cytosol from H. tubulosa coelomocytes (5-HCC) was tested against a reference group of Gram-negative and Gram-positive human pathogens. Minimal inhibitory concentrations (MICs) ranging from 125 to 500 mg/ml were determined against tested strains. The observed biological activity of 5-HCC could be due to two novel peptides, identified by capillary RP-HPLC/nESI-MS/MS, which present the common chemical-physical characteristics of antimicrobial peptides. Such peptides were c…

Espressione di galactine nell'infiammazione e nella larva natante di Ciona intestinalis

Enhanced CiTNFα expression is an inflammatory response of the ascidian Ciona intestinalis

Il sistema della profenolossidasi in Ciona intestinalis durante il processo infiammatorio indotto da LPS

The Ciona intestinalis prophenoloxidase activating system during LPS inflammatory reaction

Physiological and agonistic behavioural response of Procambarus clarkii to an acoustic stimulus

Summary This study examined the effects of an acoustic stimulus on the haemolymph and agonistic behaviour of the red swamp crayfish Procambarus clarkii. The experiment was conducted in a tank equipped with a video recording system using 6 groups (3 control and 3 test groups) of five adult crayfish (30 specimens in total). After one hour of habituation, the behaviour of the crayfish was monitored for two hours. During the second hour, the animals in the test groups were exposed to a linear sweep (frequency range 0.1-25 kHz; peak amplitude 148 dBrms re 1 µPa at 12 kHz) acoustic stimulus for 30 minutes. Exposure to the noise produced significant variations in haemato-immunological parameters a…

FACIT collagen (1α-chain) is expressed by hemocytes and epidermis during the inflammatory response of the ascidian Ciona intestinalis

Based on previous cloning and sequencing study, real-time PCR and in situ hybridization assays of the inflamed body wall of LPS-injected Ciona intestinalis showed the enhanced gene expression of a collagen with FACIT structural features (Ci-type IX-Col 1alpha-chain). By using specific antibodies raised against an opportunely chosen Ci-type IX-Col synthetic peptide, the fibroblast property of hemocytes challenged in vitro with LPS (at 4h) was displayed by flow cytometry, while immunocytochemistry identified hemocytes with large granules (morula cells) as collagen-producing cells. Hemocyte lysate supernatant analyzed in immunoblotting contained a 60 kDa band identifiable as 1alpha-chain-Ci-ty…

Collective Locomotion of Human Cells, Wound Healing and Their Control by Extracts and Isolated Compounds from Marine Invertebrates

The collective migration of cells is a complex integrated process that represents a common theme joining morphogenesis, tissue regeneration, and tumor biology. It is known that a remarkable amount of secondary metabolites produced by aquatic invertebrates displays active pharmacological properties against a variety of diseases. The aim of this review is to pick up selected studies that report the extraction and identification of crude extracts or isolated compounds that exert a modulatory effect on collective cell locomotion and/or skin tissue reconstitution and recapitulate the molecular, biochemical, and/or physiological aspects, where available, which are associated to the substances und…

Stress and immunomodulation indicators in gilthead seabream (Sparus aurata): preliminary data concerning the alterations due to the social behaviour.

Transcriptomic Analyses Reveal 2 and 4 Family Members of Cytochromes P450 (CYP) Involved in LPS Inflammatory Response in Pharynx of Ciona robusta

Cytochromes P450 (CYP) are enzymes responsible for the biotransformation of most endogenous and exogenous agents. The expression of each CYP is influenced by a unique combination of mechanisms and factors including genetic polymorphisms, induction by xenobiotics, and regulation by cytokines and hormones. In recent years, Ciona robusta, one of the closest living relatives of vertebrates, has become a model in various fields of biology, in particular for studying inflammatory response. Using an in vivo LPS exposure strategy, next-generation sequencing (NGS) and qRT-PCR combined with bioinformatics and in silico analyses, compared whole pharynx transcripts from naïve and LPS-exposed C. robusta…

Il cirripede Pollicipes pollicipes una prelibatezza galiziana

Prophenoloxidase system is activated in the tunic inflammatory response of ciona intestinalis

Haemolymphatic Parameters in Two Aquaculture Crustacean Species Cherax destructor (Clark, 1836) and Cherax quadricarinatus (Von Martens, 1868)

In the last few years, there has been a notable development in the breeding of freshwater shrimp (astaciculture), which involved various species and in particular, the two Australian Parastacidae species, Cherax destructor and Cherax quadricarinatus. Information about the haemolymphatic parameters of these two species is fragmentary, and filling these gaps becomes important given their importance in aquaculture. Cellular and biochemical parameters were analyzed in both species to create a reference baseline for these parameters to identify the state of welfare or suffering of these animals. The results showed that the total haemocyte count, haemocyte subpopulations, enzymatic activities and…

Inflammation in ascidians

Seabream(Sparus aurata)hierarchy among alfa and beta subordinates and dominant interplay affects stress responses and phagocytic activity by peritoneal cavity cells

seabream, subordinates, dominant

Indicatori di stress ed immunomodulazione in Sparus aurata: dati preliminari sulle modificazioni indotte dal comportamento sociale di sottomissione.

Haemolymphatic parameters in two aquaculture crustacean species Cherax destructor (Clark, 1836) and Cherax quadricarinatus (von Martens, 1868)

The growing demand for animal proteins of aquatic origin such as fish, molluscs and crustaceans have prompted some European and Italian companies to focus their attention on some species of freshwater crustaceans such as Cherax destructor and Cherax quadricarinatus. They are among the largest freshwater decapods, matures early, females can lay over a thousand eggs in a single brood and has a broad environmental tolerance. All these characteristics make it highly appreciated species for aquaculture and few years ago their breeding has spread to Europe and recently also in Italy (Sicily). Although some commercial yabby farms have been studied these species from different points of view (dieta…

Basic knowledge of social hierarchies and physiological profile of reared sea bass Dicentrarchus labrax (L.).

The effects of social hierarchies (dominant/subordinate individuals), such as aggressiveness, feeding order, and territoriality, are some of the characteristics used for describing fish behaviour. Social hierarchy patterns are still poorly understood in European-reared sea bass (Dicentrarchus labrax). In this work, we examine the social interactions among captive fish integrating behavioural and physiological profiles. Groups of three fish with EMG (electromyogram) radio transmitters were monitored for two weeks via video recording. Plasma levels of cortisol, glucose, lactate and lysozyme as well as haematological parameters such as haemoglobin, haematocrit and RBCC (red blood cell count) w…

Allograft Inflammatory Factor AIF-1: early immune response in the Mediterranean sea urchin Paracentrotus lividus

Echinoderms are a phylum of deuterostomic invertebrates that play a key role in maintaining the biodiversity of marine ecosystems. They represent a good study model for immunity because their coelomic fluid contains different types of cells involved in the inflammatory response: the coelomocytes. In the case of the sea urchin Paracentrotus lividus, the coelomocyte population is mainly represented by amoebocytes and uncoloured spherulocytes that implement a defence program through phagocytosis, encapsulation, cytotoxicity, and production of antimicrobial agents. The purpose of this study was to find evidence of a possible modulating effect of lipopolysaccharide LPS on the expression of the A…

Molecules and cells in inflammatory responses of the ascidian Ciona intestinalis

Expression of a glucocorticoid receptor (DlGR1) in several tissues of the teleost fish Dicentrarchus labrax

Since glucocorticoids have a role in maintaining the homeostatic status in fish, in the present paper mRNA expression (in situ hybridization) and tissue immunohistochemical localization of a glucocorticoid receptor (DlGR1) in several Dicentrarchus labrax organs are reported. Riboprobe and specific antibodies were prepared by using the DlGR1 that has been previously cloned and sequenced from peritoneal cavity leukocytes. Both mRNA and receptor were identified in head kidney, spleen, gills, intestine, heart and liver tissues. The functional roles of DlGR1 localization are discussed.

The use of an integrative approach to identify coelomocytes in three species of the genus Holothuria (Echinodermata)

Coelomocytes in the Holothuroidea are traditionally identified according to their morphology through light, fluorescence, or electron microscopy. Former studies have typically used only one method, with few works combining two or more approaches. Studies using cytocentrifugation to study these cells are scarcer. Thus, for the first time, an integrative approach was used to compare coelomocytes in Holothuroidea. This approach consisted of living and stained cells, scanning electron microscopy (for spherule cells), and accurate morphometric analyses. Specifically, we used specimens of Holothuria grisea, Holothuria arenicola, and Holothuria tubulosa to test whether cytocentrifugation could be …

Confinement stress in sea bass (Dicentrarchus labrax) depresses peritoneal leukocyte cytotoxicity

Fish respond to stressful conditions via neuroendocrine responses (primary response) which result in increased levels of plasma cortisol which is considered immunosuppressive. Sea bass were confined at low (10 kg/m3) and high (60 kg/m3) density for 3-48 h. Plasma cortisol and glucose were evaluated and two principal cellular immune responses were assayed. A significant increase in plasma cortisol and glucose levels, as well as osmolarity, was found following stress. In addition, phagocytic activity, as shown by reactive oxygen species (ROS) production by challenged head kidney phagocytes and cytotoxic activity of eosinophilic granule cells from peritoneal cavity against K562 tumour cell lin…

Effects of acoustic stimulation on biochemical parameters in the digestive gland of Mediterranean mussel Mytilus galloprovincialis (Lamarck, 1819)

Underwater sounds generated by anthropogenic activity can cause behavior changes, temporary loss of hearing, damage to parts of the body, or death in a number of marine organisms and can also affect healing and survival. In this study, the authors examined the effects of high-frequency acoustic stimulations on a number of biochemical parameters in the Mediterranean mussel, Mytilus galloprovincialis. During the experiment, animals were placed in a test tank and exposed to acoustic signals [a linear sweep ranging from 100 to 200 kHz and lasting 1 s, with a sound pressure level range of between 145 and 160 dBrms (re 1μParms)] for 3 h. Total haemocyte count was assessed and glucose levels, cyto…

Calibrating accelerometer data, as a promising tool for health and welfare monitoring in aquaculture: Case study in European sea bass (Dicentrarchus labrax) in conventional or organic aquaculture

Abstract Both the conventional and organic aquaculture sectors have grown rapidly over the past few years. Moreover, welfare has attracted increased attention on the part of both consumers and governments. However, fish welfare assessment is complex and thus needs to adapt measurements that are easily applicable to aquaculture conditions. In this study, in addition to classical welfare indicator measurements (physiological stress indicators and growth performance), we recorded the swimming activity data using acoustic transmitters to evaluate the welfare of European sea bass (Dicentrarchus labrax) fed a conventional vs. an organic diet. Prior feeding trial, the swimming activity recorded by…

Modulazione cortisolo-dipendente delle Hsp 70 e 90 nel pronefro di Dicentrarchus labrax in vivo ed in vitro

La risposta infiammatoria in Ciona intestinalis è caratterizzata dall’aumento di lectine specifiche per D-galattosidi e dall’espressione di collagene FACIT di tipo IX

AQUEOUS EXTRACTS FROM LEAVES AND RHIZOMES OF THE MARINE SEAGRASS POSIDONIA OCEANICA EXHIBIT ANTI-LIVER CANCER ABILITY IN VITRO

Comparative analysis of fucose binding lectins isolated and characterized from different teleost species, and distribution of a F-Lectin during Dicentrarchus labrax ontogenesis

Underwater noise from boats: Measurement of its influence on the behaviour and biochemistry of the common prawn (Palaemon serratus, Pennant 1777)

Abstract This study examined the effects of recorded boat noises on the behaviour and biochemistry of the common prawn ( Palaemon serratus ) in laboratory settings. The experiment was carried out in a tank equipped with a video-recording system using six groups (three control and three tested) of eight common prawns (48 animals in total). After habituation for 1 h, the behaviour of the prawns was monitored for 1 h. During the last 30 min, the animals in the test groups were exposed to noise resembling a marine area with high anthropogenic acoustic pollution. The exposure to noise produced significant changes in locomotor patterns, presence inside or outside a shelter, total protein concentr…

BROILER CHICKENS AND THEIR HEALTH STATUS IN FARM: WINE WASTE, AN ADDED VALUE ON FEEDING

Various key economic sectors of the National and Sicilian economy, in addition to contributing to the economic growth also cause a considerable production of waste which today are a problem both for companies then for the environment. For this reason, there is considerable interest in this waste and in the possibility of their reuse to extract bioactive molecules with added value, all in accordance with the objectives of the Blue Economy. In this regard, it has already been demonstrated that from the production waste of various economic sectors it is possible to obtain bioactive molecules with anticancer, antimicrobial, cosmetic, antioxidant and nutraceutical properties (Mauro et al., 2022)…

Effetti del cadmio sulla fagocitosi e sull'attività di formazione di placche di lisi dei celomociti di Parcentrotus lividus

Molecular and physiological characterization of in vivo Sulfamethoxazole response in Procambarus clark

Preliminare caratterizzazione di un’attività citotossica nel supernatante del lisato degli emociti con granulo rifrangente di Ciona intestinalis

Evolution of Innate Immunity. Components of inflammatory reaction in Ciona intestinalis

Localization of glucocorticoid receptor DIGR1 in tissues pf teleost Dicentrarchus labrax.

Transcriptomic and Bioinformatic Analyses Identifying a Central Mif-Cop9-Nf-kB Signaling Network in Innate Immunity Response of Ciona robusta

The Ascidian C. robusta is a powerful model for studying innate immunity. LPS induction activates inflammatory-like reactions in the pharynx and the expression of several innate immune genes in granulocyte hemocytes such as cytokines, for instance, macrophage migration inhibitory factors (CrMifs). This leads to intracellular signaling involving the Nf-kB signaling cascade that triggers downstream pro-inflammatory gene expression. In mammals, the COP9 (Constitutive photomorphogenesis 9) signalosome (CSN) complex also results in the activation of the NF-kB pathway. It is a highly conserved complex in vertebrates, mainly engaged in proteasome degradation which is essential for maintaining proc…

Seasonal trend of Anisakidae infestation in South Mediterranean bluefish

A total of 1104 fish samples from markets of Sicily were analysed for the detection and species identification of Anisakidae nematodes. The preliminary analysis of the fish samples showed the presence of 2459 larvae. All the fish species revealed different prevalence of infestation, with a maximum of 100% for Lepidopus caudatus and a minimum of 4.5% in Sardina pilchardus. The 80% of the larvae examined by PCR-RFLP analysis belonged to Anisakis pegreffii species. The seasonal infestation trend of Anisakis was evaluated in all the fish sample examined. The results of the seasonal infestation trend showed a marked connection with the ecological aspects of the fish species examined. As far as w…

Espressione di DlGR1 nell'intestino di D.labrax dopo trattamento in vivo con cortisolo

Espression of a type IX-like collagen in tissue injury of the ascidian Ciona intestinalis during inflammatory process

Enhanced expression of a CinTNF gene in the LPS challenged inflammatiry responses of the ascidian Ciona intestinalis

Effects of cadmium on expression of the hsp70 in sea bass (Dicentrarchus labrax L., Osteichthyes. Moronidae) blood cells

Abstract To gain further insight into the role of cortisol in Wsh innate immune responses, we cloned and sequenced a 2592 bp cDNA from sea bass (Dicentrarchus labrax) peritoneal leukocytes (PCLs) encoding a glucocorticoid receptor (DlGR1). The deduced aminoacid sequence displayed that DlGR1 belong to a multigenic family of steroid hormone receptors, and exhibited high homology (80%) to the Burton’s mouth breeder (Haplochromis burtoni) HbGR1. The DlGR1 functional domains presented homologies with those of several vertebrate species. In situ hybridization assay revealed that DlGR1 was expressed in macrophages and neutrophils from the peritoneal cavity. Since in a previous paper, sea bass PCL …

Inflammatory response of Ciona intestinalis involve enhanced D-galactoside-specific lectins and FACIT collagen expression.

Inducible lectins with galectin properties and human IL1alpha epitopes opsonize yeast during the inflammatory response of the ascidian Ciona intestinalis.

Studies on inducible ascidian lectins may shed light on the evolutionary emergence of cytokine functions. Here, we show that the levels of opsonins, with IL1alpha-epitopes, increase in Ciona intestinalis hemolymph as a response to an inflammatory stimulus and, in particular, to intratunic injection of lipopolysaccharide (LPS). The inflammatory agent promptly (within 4 h) enhances Ca(2+)-independent serum hemagglutinating and opsonizing activities, which are both inhibited by D-galactose and D-galactosides (alpha-lactose, N-acetyl-D-lactosamine, thio-digalactoside), suggesting that anti-rabbit erythrocyte lectins with galectin properties are involved as opsonins. Inducible galectin molecules…

Effect of cadmium exposure on phagocytosis and plaque lysis activity of Paracentrotus lividus coelomocyte

Anche i pesci si stressano