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1.
Mar Drugs ; 22(4)2024 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-38667789

RESUMO

Sea anemones are valuable for therapeutic research as a diversified source of bioactive molecules, due to their diverse bioactive molecules linked to predation and defence mechanisms involving toxins and antimicrobial peptides. Acid extracts from Actinia equina tentacles and body were examined for antibacterial activity against Gram-positive, Gram-negative bacteria, and fungi. The peptide fractions showed interesting minimum inhibitory concentration (MIC) values (up to 0.125 µg/mL) against the tested pathogens. Further investigation and characterization of tentacle acid extracts with significant antimicrobial activity led to the purification of peptides through reverse phase chromatography on solid phase and HPLC. Broad-spectrum antimicrobial peptide activity was found in 40% acetonitrile fractions. The resulting peptides had a molecular mass of 2612.91 and 3934.827 Da and MIC ranging from 0.06 to 0.20 mg/mL. Sequencing revealed similarities to AMPs found in amphibians, fish, and Cnidaria, with anti-Gram+, Gram-, antifungal, candidacidal, anti-methicillin-resistant Staphylococcus aureus, carbapenemase-producing, vancomycin-resistant bacteria, and multi-drug resistant activity. Peptides 6.2 and 7.3, named Equinin A and B, respectively, were synthesized and evaluated in vitro towards the above-mentioned bacterial pathogens. Equinin B exerted interesting antibacterial activity (MIC and bactericidal concentrations of 1 mg/mL and 0.25 mg/mL, respectively) and gene organization supporting its potential in applied research.


Assuntos
Testes de Sensibilidade Microbiana , Animais , Antibacterianos/farmacologia , Antibacterianos/isolamento & purificação , Antibacterianos/química , Peptídeos Antimicrobianos/farmacologia , Peptídeos Antimicrobianos/isolamento & purificação , Peptídeos Antimicrobianos/química , Anêmonas-do-Mar/química , Bactérias Gram-Positivas/efeitos dos fármacos , Bactérias Gram-Negativas/efeitos dos fármacos , Anti-Infecciosos/farmacologia , Anti-Infecciosos/isolamento & purificação , Anti-Infecciosos/química , Fungos/efeitos dos fármacos
2.
Int J Mol Sci ; 24(10)2023 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-37240205

RESUMO

Benthic marine invertebrates, such as corals, are often subjected to injury caused by several sources. Here, the differences and characteristics in injured and health tissues in terms of cellular components are shown through a histological investigation of the soft coral Anemonia viridis at 0 h, 6 h, 24 h, and 7 days after injury caused by tentacle amputation. In addition, a new tool was used for the first time in invertebrates, positron emission tomography, in order to investigate the events that occur during regeneration within a longer time period (0 h, 24 h, and 14 days after the tentacles were cut). Higher integrated density values were measured through a densitometric analysis in sections stained with Fontana-Masson at 24 h after the tentacles were cut. This suggests an increase in melanin-like containing cells and a subsequent increase in fibroblast-like cells differentiated by amoebocytes that converge to the lesion site in the early stages of inflammation and regeneration. This work provides, for the first time, an elucidation of the events that occur during wound-healing and regeneration in basal metazoan, focusing on the characterisation of immune cells and their role. Our results indicate that Mediterranean anthozoan proves to be a valuable model for studying regeneration. Many events highlighted in this research occur in different phyla, suggesting that they are highly conserved.


Assuntos
Antozoários , Anêmonas-do-Mar , Animais , Invertebrados , Organismos Aquáticos , Cicatrização
3.
Int J Mol Sci ; 23(12)2022 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-35743284

RESUMO

The aim of this Special Issue is to highlight the close functional and highly conserved link between innate immunity, homeostasis maintenance, inflammation, tissue remodeling and regeneration [...].


Assuntos
Imunidade Inata , Inflamação , Animais , Homeostase , Invertebrados
4.
Fish Shellfish Immunol ; 119: 384-396, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34687879

RESUMO

Ascidians are marine invertebrate chordates belonging to the earliest branch (Tunicata) in the chordate phylum, therefore, they are of interest for studying the evolution of immune systems. Due to the known genome, the non-colonial Ciona robusta, previously considered to be C. intestinalis type A, is a model species for the study of inflammatory response. The internal defense of ascidians mainly relies on hemocytes circulating in the hemolymph and pharynx. Hemocytes can be in vivo challenged by LPS injection and various granulocyte and vacuolated cell populations differentiated to produce and release inflammatory factors. Molecular biology and gene expression studies revealed complex defense mechanisms involving different inflammatory hemocytes. Furthermore, cloning procedures allowed sequence analyses and molecular studies disclose immune-related gene families including TOLL-like receptors, galectins, C-type lectins, collectins, interlectins, pentraxine-like, peroxinectins, complement factors-like, TNFα-like, IL-17-like, TGF-like, MIF-like. These genes are promptly upregulated by the inflammatory stimulus and show a time course of transcription similar to each other. Domains sequence similarity and phylogenetic relationships with the vertebrate counterparts are shedding some light on immune-related gene evolution. Selective bioassays as well as bioinformatic approaches have allowed the characterization of antimicrobial peptides and the identification of post transcriptional molecular mechanisms able of influencing dynamics of gene regulation are described. In synthesis, the purpose of this article is to further explore the topic of hemocyte and molecules related to internal defence of ascidians involved in the inflammatory reaction, as well as to discuss current and future study options through a detailed literature review.


Assuntos
Ciona intestinalis , Animais , Peptídeos Antimicrobianos , Ciona intestinalis/imunologia , Hemócitos , Lipopolissacarídeos , Filogenia
5.
Fish Shellfish Immunol ; 114: 330-339, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34015481

RESUMO

The external mucus layer that covers fish skin contains numerous immune substances scarcely studied that act as the first line of defence against a broad spectrum of pathogens. This study aimed to characterize and describe for the first time several humoral immune defence parameters in the skin mucus of the European eel (Anguilla anguilla) after intraperitoneal injection with Vibrio anguillarum or Tenacibaculum soleae. This study evaluated several immune-related enzymes and bactericidal activity against fish pathogenic bacteria in the skin mucus of European eels at 24, 48, and 72 h post-challenge. The results demonstrated that European eel skin mucus showed significant increments in peroxidase and lysozyme activity at 48 and 72 h after V. anguillarum challenge, compared to other experimental groups. In the case of antiprotease activity, an increase was observed at 24 h in the skin mucus of fish challenged with V. anguillarum compared to unchallenged fish, while this activity was undetected at 48 and 72 h. In contrast, protease activity had decreased at 48 and 72 h in the skin mucus of fish challenged with V. anguillarum compared to the unchallenged group. Regarding bactericidal activity, a high growth capacity of T. soleae was observed in the skin mucus of all experimental groups. Interestingly, the skin mucus from fish challenged with V. anguillarum exhibited increased bactericidal activity against this bacterium at 48 h, compared to unchallenged fish. Finally, severe histopathological alterations were observed in the gills and liver at the end of the trial (72 h), whereas the skin showed only an overspread presence of goblet cells in the challenged fish compared to unchallenged fish. The present results may give new insights into the mucosal immune system of this primitive species with potential applications in aquaculture.


Assuntos
Anguilla , Doenças dos Peixes/microbiologia , Infecções por Flavobacteriaceae/veterinária , Imunidade nas Mucosas/fisiologia , Tenacibaculum , Vibrio , Animais , Infecções por Flavobacteriaceae/tratamento farmacológico , Infecções por Flavobacteriaceae/microbiologia , Pele/imunologia , Vibrioses/imunologia , Vibrioses/microbiologia , Vibrioses/veterinária
6.
Int J Mol Sci ; 22(11)2021 May 31.
Artigo em Inglês | MEDLINE | ID: mdl-34073146

RESUMO

Given the anatomical simplicity and the extraordinary ability to regenerate missing parts of the body, Cnidaria represent an excellent model for the study of the mechanisms regulating regenerative processes. They possess the mesoglea, an amorphous and practically acellular extracellular matrix (ECM) located between the epidermis and the gastrodermis of the body and tentacles and consists of the same molecules present in the ECM of vertebrates, such as collagen, laminin, fibronectin and proteoglycans. This feature makes cnidarians anthozoans valid models for understanding the ECM role during regenerative processes. Indeed, it is now clear that its role in animal tissues is not just tissue support, but instead plays a key role during wound healing and tissue regeneration. This study aims to explore regenerative events after tentacle amputation in the Mediterranean anemone Anemonia viridis, focusing in detail on the reorganization of the ECM mesoglea. In this context, both enzymatic, biometric and histological experiments reveal how this gelatinous connective layer plays a fundamental role in the correct restoration of the original structures by modifying its consistency and stiffness. Indeed, through the deposition of collagen I, it might act as a scaffold and as a guide for the reconstruction of missing tissues and parts, such as amputated tentacles.


Assuntos
Matriz Extracelular/metabolismo , Regeneração , Anêmonas-do-Mar/crescimento & desenvolvimento , Cicatrização , Animais , Colágeno Tipo I/metabolismo
7.
Fish Shellfish Immunol ; 72: 452-458, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29146447

RESUMO

Riboprobes (in situ hybridization) and antibodies (immunohistochemistry), previously used to show the upregulation of Ciona intestinalis (Type A) galectins (CiLgals-a, CiLgals-b) and phenoloxidase (CinPO2) immune-related genes, were tested on histological sections of the ovary. The ovarian follicles are composed of oocytes encased by follicular cells (FCs) and test cells (TCs). Results show the transcription upregulation of both CiLgals and CinPO2 genes in the vitellogenic FCs, conversely distinct cytolocalization of the proteins are shown. At vitellogenic stage, the CiLgals are localized in the FCs, in the oocyte cytoplasm, and close to the germinal vesicle (GV), whereas the CinPO2 was never identified in the FCs. In a presumptive advanced phase and at the post-vitellogenic stage the TCs appear to be labelled by the CinPO2 riboprobe, and the protein identified by the antibody suggesting an mRNA transcytosis process from FCs. At post-vitellogenic stage the CiLgals mainly enrich the GV nucleoplasm, whereas the CinPO2 is contained in TCs and in the ooplasm but never found in the GV. This finding sheds new light on a former paper in which TCs were reported to be the only CinPO2-producing cells in the ovarian follicle. Finally, CiLgals and CinPO2 genes transcription and proteins production seem to be associated with accessory cells during their differentiation from vitellogenic to post-vitellogenic stage. The present findings promote further research on the early upregulation of immune-related genes, and the potential multifunctional role of the produced proteins. In addition further insight on the accessory cells involvement in ascidian oogenesis are reported.


Assuntos
Ciona intestinalis/genética , Galectinas/genética , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Monofenol Mono-Oxigenase/genética , Animais , Ciona intestinalis/imunologia , Feminino , Galectinas/imunologia , Perfilação da Expressão Gênica , Genes , Monofenol Mono-Oxigenase/imunologia , Ovário/metabolismo
8.
Mar Drugs ; 16(9)2018 Aug 23.
Artigo em Inglês | MEDLINE | ID: mdl-30142922

RESUMO

The phylum Cnidaria is an ancient branch in the tree of metazoans. Several species exert a remarkable longevity, suggesting the existence of a developed and consistent defense mechanism of the innate immunity capable to overcome the potential repeated exposure to microbial pathogenic agents. Increasing evidence indicates that the innate immune system in Cnidarians is not only involved in the disruption of harmful microorganisms, but also is crucial in structuring tissue-associated microbial communities that are essential components of the Cnidarian holobiont and useful to the animal's health for several functions, including metabolism, immune defense, development, and behavior. Sometimes, the shifts in the normal microbiota may be used as "early" bio-indicators of both environmental changes and/or animal disease. Here the Cnidarians relationships with microbial communities and the potential biotechnological applications are summarized and discussed.


Assuntos
Cnidários/fisiologia , Imunidade Inata , Microbiota/imunologia , Simbiose/imunologia , Animais , Biotecnologia/métodos , Cnidários/microbiologia , Homeostase/imunologia , Mucosa/imunologia , Mucosa/microbiologia , Filogenia
9.
Fish Shellfish Immunol ; 45(1): 112-23, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25700783

RESUMO

Aquatic animals are continuously exposed to chemical pollutants but the effects evoked in skin surfaces, which receive the most direct contact with them, are poorly investigated. Terminal carbohydrate composition and immunological components present in skin mucus of gilthead seabream (Sparus aurata L.) specimens exposed to waterborne sublethal dosages of heavy metals [arsenic (As2O3), cadmium (CdCl2) and mercury (CH3HgCl) at 5, 5 and 0.04 µM, respectively for 2, 10 and 30 days were analysed. Moreover, the presence of a fucose binding lectin (FBL) was evaluated by western blot and the protein profiles were by SDS-PAGE and HPLC. Results showed little effects of heavy metals in the presence of several terminal carbohydrates with few increments or decrements. Most of the enzyme activities related to immune responses were increased upon heavy metal exposure in the skin mucus including bactericidal activity. Methylmercury produced the most dramatic changes increasing all the activities. Moreover, the FBL was undetected in any of the control fish skin mucus but was evident in all the heavy metal exposed fish. In addition, As and Cd produced a clear change in the protein profile as evidenced by the lack of a protein band of around 12 kDa which is absent. These protein changes were more evident with the HPLC study showing the presence of different peaks and differences in intensity. The present results could be useful for better understanding the role and their behaviour of the mucosal immunity in skin as a key component of the innate immune system against pollutants.


Assuntos
Imunidade Inata/efeitos dos fármacos , Imunidade nas Mucosas/efeitos dos fármacos , Metais Pesados/toxicidade , Perciformes/imunologia , Poluentes Químicos da Água/toxicidade , Animais , Distribuição Aleatória , Pele/imunologia
10.
Mar Drugs ; 13(8): 5276-96, 2015 Aug 19.
Artigo em Inglês | MEDLINE | ID: mdl-26295400

RESUMO

The mucus produced by many marine organisms is a complex mixture of proteins and polysaccharides forming a weak watery gel. It is essential for vital processes including locomotion, navigation, structural support, heterotrophic feeding and defence against a multitude of environmental stresses, predators, parasites, and pathogens. In the present study we focused on mucus produced by a benthic cnidarian, the sea anemone Actinia equina (Linnaeus, 1758) for preventing burial by excess sedimentation and for protection. We investigated some of the physico-chemical properties of this matrix such as viscosity, osmolarity, electrical conductivity, protein, carbohydrate, and total lipid contents. Some biological activities such as hemolytic, cytotoxic, and antibacterial lysozyme-like activities were also studied. The A. equina mucus is mainly composed by water (96.2% ± 0.3%), whereas its dry weight is made of 24.2% ± 1.3% proteins and 7.8% ± 0.2% carbohydrates, with the smallest and largest components referable to lipids (0.9%) and inorganic matter (67.1%). The A. equina mucus matrix exhibited hemolytic activity on rabbit erythrocytes, cytotoxic activity against the tumor cell line K562 (human erythromyeloblastoid leukemia) and antibacterial lysozyme-like activity. The findings from this study improve the available information on the mucus composition in invertebrates and have implications for future investigations related to exploitation of A. equina and other sea anemones' mucus as a source of bioactive compounds of high pharmaceutical and biotechnological interest.


Assuntos
Antozoários/metabolismo , Fatores Biológicos/farmacologia , Venenos de Cnidários/farmacologia , Muco/metabolismo , Anêmonas-do-Mar/metabolismo , Animais , Antozoários/química , Antibacterianos/química , Antibacterianos/farmacologia , Fatores Biológicos/química , Fatores Biológicos/metabolismo , Carboidratos/química , Linhagem Celular Tumoral , Venenos de Cnidários/química , Citotoxinas/química , Citotoxinas/farmacologia , Eritrócitos/efeitos dos fármacos , Hemolíticos/química , Hemolíticos/farmacologia , Humanos , Invertebrados/química , Invertebrados/metabolismo , Células K562 , Muco/química , Coelhos , Anêmonas-do-Mar/química
11.
Sci Total Environ ; 927: 172177, 2024 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-38575005

RESUMO

The rise of plastic production has triggered a surge in plastic waste, overwhelming marine ecosystems with microplastics. The effects of climate change, notably changing salinity, have shaped the dynamics of coastal lagoons. Thus, understanding the combined impact of these phenomena on marine organisms becomes increasingly crucial. To address these knowledge gaps, we investigated for the first time the interactive effects of environmental microplastics (EMPs) and increased salinity on the early development of Mytilus galloprovincialis larvae. Morphological assessments using the larval embryotoxicity test revealed larval anomalies and developmental arrests induced by EMPs and increased salinity. Transcriptomic analyses targeting 12 genes involved in oxidative stress, apoptosis, DNA repair, shell formation, and stress proteins were conducted on D-larvae uncovered the potential effects of EMPs on shell biomineralization, highlighting the role of Histidine Rich Glycoproteine (HRG) and tubulin as crucial adaptive mechanisms in Mytilus sp. in response to environmental shifts. Furthermore, we explored oxidative stress and neurotoxicity using biochemical assays. Our findings revealed a potential interaction between EMPs and increased salinity, impacting multiple physiological processes in mussel larvae. Our data contribute to understanding the cumulative effects of emerging anthropogenic pollutants and environmental stressors, emphasizing the need for a holistic approach to assessing their impact on marine ecosystems.


Assuntos
Larva , Microplásticos , Mytilus , Poluentes Químicos da Água , Animais , Mytilus/efeitos dos fármacos , Mytilus/fisiologia , Poluentes Químicos da Água/toxicidade , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Microplásticos/toxicidade , Estresse Salino/efeitos dos fármacos , Estresse Oxidativo , Salinidade
12.
Animals (Basel) ; 14(15)2024 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-39123695

RESUMO

The close phylogenetic relationship between ascidians (Tunicata) and vertebrates makes them a powerful model for studying the innate immune system. To better understand the nature and dynamics of immune responses and the mechanisms through which bacterial infections are detected and translated into inflammation in Ciona robusta, we applied an approach combining in vivo lipopolysaccharide (LPS) stimulation, immune-labelling techniques and functional enzymatic analyses. The immunohistochemistry showed that Toll-like receptor 4 (TLR4) and nuclear factor kappa B (NFκB) were expressed during the inflammatory pharynx response 4 h post-LPS, with the formation of nodules in pharynx vessel lumen. Also, the endothelium vessels were involved in the inflammatory response. Observations of histological sections from naive and buffer-inoculated ascidians confirmed an immuno-positive response. Enzyme immune parameters-which included the activity of phenoloxidase, glutathione peroxidase, lysozyme, alkaline phosphatase and esterase-showed up-modulation 4 h after LPS injection, confirming their participation during ascidian inflammatory response. These findings provide new insights into the mechanisms underlying the LPS-induced C. robusta response and suggest that a broad innate immune mechanism, as in vertebrates, is involved in the regulation of inflammatory responses. Further findings in this direction are needed to cover knowledge gaps regarding the organized set of molecular and cellular networks involved in universal immune interactions with pathogens.

13.
Antibiotics (Basel) ; 13(3)2024 Mar 02.
Artigo em Inglês | MEDLINE | ID: mdl-38534669

RESUMO

The Eurasian woodcock (Scolopax rusticola) belongs to those bird species that make systematic migratory flights in spring and autumn in search of favorable breeding and wintering areas. These specimens arrive in the Mediterranean Area from northeastern European countries during the autumn season. The purpose of this study was to assess whether woodcocks can carry antibiotic resistance genes (ARGs) along their migratory routes. Although the role of migratory birds in the spread of some zoonotic diseases (of viral and bacterial etiology) has been elucidated, the role of these animals in the spread of antibiotic resistance has not yet been clarified. In this study, we analyzed the presence of beta-lactam antibiotic resistance genes. The study was conducted on 69 strains from 60 cloacal swabs belonging to an equal number of animals shot during the 2022-2023 hunting season in Sicily, Italy. An antibiogram was performed on all strains using the microdilution method (MIC) and beta-lactam resistance genes were investigated. The strains tested showed no phenotypic resistance to any of the 13 antibiotics tested; however, four isolates of Enterobacter cloacae and three of Klebsiella oxytoca were found to carry the blaIMP-70, blaVIM-35, blaNDM-5 and blaOXA-1 genes. Our results confirm the importance of monitoring antimicrobial resistance among migratory animals capable of long-distance bacteria spread.

14.
Artigo em Inglês | MEDLINE | ID: mdl-36182080

RESUMO

The aim of this work is to study the immune responses of the polychaete Sabella spallanzanii after exposure to copper sulphate, an immunomodulating agent in marine organisms, and the multiple stresses caused by Escherichia coli infection, to validate the species as a model organism in marine-coastal biomonitoring programmes. Polychaetes were housed in laboratory and divided into five experimental groups: 1. Control (no microinjected), 2. filtered seawater + TBS injection (control of point 3), 3. filtered seawater + E. coli injection (control of point 4), 4. CuSO4 + TBS injection (control of point 5), and 5. CuSO4 + E. coli injection. The immune variables, esterase and alkaline phosphatase activity, cytotoxicity and detoxifying/antioxidant enzymes such as glutathione peroxidase were evaluated in total body extracts of the animals. Moreover, toll-like receptor, allograft inflammatory factor-1, lysozyme and haemagglutinating activity were investigated to highlight possible interactions. Indeed, the results of this work demonstrate the immunomodulating effect of copper sulphate on S. spallanzanii total body extracts related to oxidative stress and inflammatory markers.


Assuntos
Sulfato de Cobre , Poliquetos , Animais , Sulfato de Cobre/toxicidade , Escherichia coli , Água do Mar , Estresse Oxidativo
15.
Fish Shellfish Immunol ; 33(3): 641-4, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22750680

RESUMO

The antifungal peptide mytimycin (MytM) is synthesized by hemocytes of the Mediterranean mussel, Mytilus galloprovincialis. In addition to sequence and gene structure diversities previously reported from pooled hemocytes, the present report focused on the expression of mytm gene in individual M. galloprovincialis, before and after challenge. Within untreated mussel, MytM mRNA was observed by ISH in about 42% of circulating hemocytes, characterized by large, diffuse nucleus. Injection with Fusarium oxysporum increased such percentage, but in only some of the mussels. Similarly, MytM gene expression increased after injection in only some of the mussels, as measured by qPCR. Responders and not responders are common evidence in any given population of organisms. Nevertheless, even if the use of proper pool size selection has been practised to find out and evaluate the most common response trends, individual analyses must be regarded as optimal.


Assuntos
Peptídeos Catiônicos Antimicrobianos/metabolismo , Regulação da Expressão Gênica , Mytilus/metabolismo , Mytilus/microbiologia , Animais , Fusarium/fisiologia , Perfilação da Expressão Gênica , Hemócitos/metabolismo , Hibridização In Situ , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real
16.
Fish Shellfish Immunol ; 32(1): 45-50, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22037382

RESUMO

We previously reported the nucleotide sequences and diversity of mytimycin (MytM) from the Mediterranean mussel, Mytilus galloprovincialis. Using real-time PCR (q-PCR), we observed that the MytM gene was mainly expressed in circulating hemocytes and to a less extent in the mantle. In vivo challenge with bacteria or with the yeast, Candida albicans, did not increase the expression as measured by q-PCR in hemocytes. By contrast, injection of the filamentous fungus, Fusarium oxysporum, induced a sudden and strong increase of expression at 9h p.i. (stimulation index of 25.7 ± 2.1). Optimum stimulating dose was 10(4) spores of F. oxysporum per mussel. In the same samples, AMP mytilin and myticin showed no stimulation. Consequently, we hypothesized the existence of 2 different signal transduction pathways, one activated by bacteria and yeast, the other triggered by filamentous fungi. A second challenge performed with F. oxysporum 24 h after the first challenge induced an increase of MytM gene expression (stimulation index of 3.5 ± 1.7). However, this second increase was significantly lower than the first, suggesting less efficient response rather than significant protection.


Assuntos
Peptídeos Catiônicos Antimicrobianos/metabolismo , Regulação da Expressão Gênica , Mytilus/genética , Mytilus/microbiologia , Adjuvantes Imunológicos/farmacologia , Animais , Peptídeos Catiônicos Antimicrobianos/genética , Fusarium/fisiologia , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/efeitos dos fármacos , Hemócitos/metabolismo , Micrococcus/fisiologia , Mytilus/efeitos dos fármacos , Mytilus/metabolismo , Vibrio/fisiologia , beta-Glucanas/farmacologia
17.
J Comp Physiol B ; 191(1): 143-158, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-32979067

RESUMO

Filter-feeding organisms accumulate xenobiotics and other substances in their tissues. They can be useful as sentinel organisms in biomonitoring of the marine compartment. Bivalve cellular immunity is ensured by phagocytosis and cytotoxic reactions carried out by hemocytes in a network with humoral responses. These can be affected by chemical contaminants in water that can be immunosuppressors also at a low concentration increasing the sensibility to pathogens. This work is an attempt to individuate cellular markers for pollution detection, investigating the effect of methylmercury (CH3HgCl) at different concentrations on the activity and hemocyte morphology of the Mediterranean mussel, Mytilus galloprovincialis. We assessed the effect of three sub-lethal concentrations of the organometal on the cellular morphology, the efficacy of phagocytosis toward yeast cells, the alteration of the lysosomal membrane and the ability to release cytotoxic molecules. The results provide information on the alteration of hemocyte viability, modification of the morphological and cytoskeletal features and besides the cellular spreading, intrinsic ability of motile cells was used as a complementary investigation method. Exposure to the contaminant affected the percentage of phagocytosis and the phagocytosis index. Moreover, morphological and cytoskeleton alteration, caused by the pollutant, leads to reduced ability to incorporate the target and adhere to the substrate and the low ability of cells to retain neutral red could depend on the effects of methylmercury on membrane permeability. These results reinforce the use of the Mediterranean mussel as model for the evaluation of environmental quality in aquatic ecosystems integrating the novel information about hemocyte functions and morphology sensibility to organic mercury.


Assuntos
Mercúrio , Mytilus , Animais , Ecossistema , Hemócitos , Fagocitose
18.
Artigo em Inglês | MEDLINE | ID: mdl-34126205

RESUMO

The increasing availability of sequenced genomes has enabled a deeper understanding of the complexity of fish lectin repertoires involved in early development and immune recognition. The teleost fucose-type lectin (FTL) family includes proteins that preferentially bind fucose and display tandemly arrayed carbohydrate-recognition domains (CRDs) or are found in mosaic combinations with other domains. They function as opsonins, promoting phagocytosis and the clearance of microbial pathogens. The Antarctic fish Trematomus bernacchii is a Perciforme living at extremely low temperatures (-1.68 °C) which is considered a model for studying adaptability to the variability of environmental waters. Here, we isolated a Ca++-independent fucose-binding protein from the serum of T. bernacchii by affinity chromatography with apparent molecular weights of 32 and 30 kDa under reducing and non-reducing conditions, respectively. We have characterized its carbohydrate binding properties, thermal stability and potential ability to recognize bacterial pathogens. In western blot analysis, the protein showed intense cross-reactivity with antibodies specific for a sea bass (Dicentrarchus labrax) fucose-binding lectin. In addition, its molecular and structural aspects, showing that it contains two CRD-FTLs confirmed that T. bernacchii FTL (TbFTL) is a bona fide member of the FTL family, with binding activity at low temperatures and the ability to agglutinate bacteria, thereby suggesting it participates in host-pathogen interactions in low temperature environments.


Assuntos
Bactérias/metabolismo , Fucose/metabolismo , Lectinas/sangue , Lectinas/fisiologia , Perciformes/fisiologia , Sequência de Aminoácidos , Animais , Regiões Antárticas , Sequência de Bases , Lectinas/isolamento & purificação , Lectinas/metabolismo , Filogenia
19.
Mol Immunol ; 46(3): 384-92, 2009 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19054563

RESUMO

Diversity of mRNAs from mytilin B, one of the five mytilins identified in the Mediterranean mussel, Mytilus galloprovincialis, has been investigated from circulating hemocytes. One mussel expressed simultaneously two to ten different mytilin B mRNAs as observed in denaturing gradient gel electrophoresis (DGGE), defining 10 individual DGGE patterns (named A to J) within the mussels from Messina, Sicily (Italy). Three patterns accounted for 79% of the individuals whereas other patterns were found in only 2-7% of the 57 analyzed mussels. Base mutations were observed at specific locations, mainly within COOH-terminus and 3'UTR, leading to 36 nucleotide sequence variants and 21 different coding sequences (cds) segregating in two different clusters. Most of the base mutations were silent, and the number of pro-peptide variants was restricted to four. Finally, as the two amino acid replacements occurred within COOH-terminus, mature peptide from mytilin B appeared unique. Multiple sequencing of partial mytilin B gene from one mussel revealed that one to four randomly distributed mutation points occurred within intron-3. Only one sequence out of the 91 analyzed contained 16 mutation points. In addition, this sequence was the only one containing four out of the six mutation points occurring within exon-4, that code for most of the COOH-terminus domain, including the unique amino acid replacement. Statistical tests for neutrality indicated negative selection pressure on signal and mature peptide domains, but possible positive selection pressure for COOH-terminus domain.


Assuntos
Peptídeos Catiônicos Antimicrobianos/genética , Peptídeos/genética , Polimorfismo Genético , Biossíntese de Proteínas , Sequência de Aminoácidos , Animais , Peptídeos Catiônicos Antimicrobianos/química , Peptídeos Catiônicos Antimicrobianos/metabolismo , Sequência de Bases , Eletroforese em Gel de Ágar , Evolução Molecular , Éxons/genética , Regulação da Expressão Gênica , Genoma/genética , Dados de Sequência Molecular , Mytilus/genética , Fases de Leitura Aberta/genética , Peptídeos/química , Peptídeos/metabolismo , Filogenia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Seleção Genética
20.
Biology (Basel) ; 9(9)2020 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-32932829

RESUMO

Anthozoa is the most specious class of the phylum Cnidaria that is phylogenetically basal within the Metazoa. It is an interesting group for studying the evolution of mutualisms and immunity, for despite their morphological simplicity, Anthozoans are unexpectedly immunologically complex, with large genomes and gene families similar to those of the Bilateria. Evidence indicates that the Anthozoan innate immune system is not only involved in the disruption of harmful microorganisms, but is also crucial in structuring tissue-associated microbial communities that are essential components of the cnidarian holobiont and useful to the animal's health for several functions including metabolism, immune defense, development, and behavior. Here, we report on the current state of the art of Anthozoan immunity. Like other invertebrates, Anthozoans possess immune mechanisms based on self/non-self-recognition. Although lacking adaptive immunity, they use a diverse repertoire of immune receptor signaling pathways (PRRs) to recognize a broad array of conserved microorganism-associated molecular patterns (MAMP). The intracellular signaling cascades lead to gene transcription up to endpoints of release of molecules that kill the pathogens, defend the self by maintaining homeostasis, and modulate the wound repair process. The cells play a fundamental role in immunity, as they display phagocytic activities and secrete mucus, which acts as a physicochemical barrier preventing or slowing down the proliferation of potential invaders. Finally, we describe the current state of knowledge of some immune effectors in Anthozoan species, including the potential role of toxins and the inflammatory response in the Mediterranean Anthozoan Anemonia viridis following injection of various foreign particles differing in type and dimensions, including pathogenetic bacteria.

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