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Animals : An Open Access Journal From... Aug 2023is one of the major threats to world aquaculture, causing fish furunculosis and high mortality rates in cultured fish, particularly salmonids. Although spp. is a...
is one of the major threats to world aquaculture, causing fish furunculosis and high mortality rates in cultured fish, particularly salmonids. Although spp. is a thoroughly studied pathogen, little is known regarding aeromoniasis in sturgeons. After a mortality outbreak, four juvenile sturgeons () were submitted for autopsy and tissue samples were collected for histopathological and microbiological studies. The external examination revealed size heterogenicity, skin hyperpigmentation and reduced body condition of sturgeons. Within the abdominal cavity, mild hepatomegaly and splenomegaly were observed, as well as generalized organic congestion. Histology revealed severe multifocal haemorrhagic and ulcerative dermatitis, mainly localized in the dorsal and latero-ventral areas of fish. The histological study also showed moderate to severe inflammation of gills and organic lesions compatible with septicaemia. Bacterial isolates were identified as subsp. using MALDI-TOF MS and PCR. Overall, the lesions first described here are consistent with those previously reported in other cultured fish species and contribute to a better understanding of the pathogenesis of subsp. in the Siberian sturgeon, aside from providing new diagnostic tools for bacterial diseases impacting the fast-growing industry of caviar.
PubMed: 37684961
DOI: 10.3390/ani13172697 -
International Journal of Molecular... Aug 2023Turbot aquaculture production is an important economic activity in several countries around the world; nonetheless, the incidence of diseases, such furunculosis, caused...
Turbot aquaculture production is an important economic activity in several countries around the world; nonetheless, the incidence of diseases, such furunculosis, caused by the etiological agent subsp. , is responsible for important losses to this industry worldwide. Given this perspective, this study aimed to evaluate early immune responses in turbot ( L.) following infection with subsp. . For this, 72 fish were individually weighed and randomly distributed into 6 tanks in a circulating seawater system. For the bacterial challenge, half of the individuals (3 tanks with 36 individuals) were infected using a peritoneal injection with the bacterial suspension, while the other half of individuals were injected with PBS and kept as a control group. Several factors linked to the innate immune response were studied, including not only haematological (white blood cells, red blood cells, haematocrit, haemoglobin, mean corpuscular volume, mean cell haemoglobin, mean corpuscular haemoglobin concentration, neutrophils, monocytes, lymphocytes, thrombocytes) and oxidative stress parameters, but also the analyses of the expression of 13 key immune-related genes (, , , , , , , , , , , , ). No significant differences were recorded in blood or innate humoral parameters (lysozyme, anti-protease, peroxidase) at the selected sampling points. There was neither any evidence of significant changes in the activity levels of the oxidative stress indicators (catalase, glutathione S-transferase, lipid peroxidation, superoxide dismutase). In contrast, pro-inflammatory (, ), anti-inflammatory (), and innate immune-related genes () were up-regulated, while another gene linked with the lipid metabolism () was down-regulated. The results showed new insights about early responses of turbot following infection with subsp. .
Topics: Animals; Flatfishes; Immunity, Innate; Interleukin-10; Tumor Necrosis Factor-alpha
PubMed: 37629124
DOI: 10.3390/ijms241612944 -
Cells Aug 2023Serum amyloid A (SAA) proteins belong to a family of acute-phase reactants, playing an integral role in defending the organism from pathological damage. Despite a wealth...
Serum amyloid A (SAA) proteins belong to a family of acute-phase reactants, playing an integral role in defending the organism from pathological damage. Despite a wealth of data on the regulation of SAA transcripts in teleosts, there is only limited information on these proteins' abundance in fish. The aim of this study is to characterise SAA protein levels in salmonids using a newly developed antibody specific to salmonid SAA. The salmonid SAA antibody detected SAA and accurately discriminated between stimulated and control specimens from rainbow trout macrophage cell line (RTS-11) in vitro, as well as rainbow trout challenged with - or flagellin-stimulated Atlantic salmon in vivo. The presence of SAA protein was analysed in RTS-11 cell line supernatants, liver, and spleen samples using ELISA, immunoblotting, and immunohistochemistry. This study is the first to characterise SAA protein levels in salmonids in vivo and in vitro. The newly developed salmonid SAA antibody was able to discriminate between stimulated and unstimulated specimens, showing that it can be used to study the acute-phase response in salmonids with the potential to be further developed into assays to monitor and evaluate health in wild and farmed fish.
Topics: Animals; Serum Amyloid A Protein; Antibodies; Oncorhynchus mykiss; Acute-Phase Proteins; Enzyme-Linked Immunosorbent Assay
PubMed: 37626907
DOI: 10.3390/cells12162097 -
International Journal of Biological... Sep 2023Serotonergic system is involved in the regulation of physiological functions and behavioral traits including cognition, memory, aggression, stress coping, appetite and...
Serotonergic system is involved in the regulation of physiological functions and behavioral traits including cognition, memory, aggression, stress coping, appetite and immunomodulation. Serotonin exerts its functions via binding distinct serotonin receptors which are classified into 7 groups. Salmonid exhibits expanded functional gene copies due to salmonid-specific whole genome duplication. However, serotonin receptor (htr) repertoire is not fully identified in rainbow trout (Oncorhynchus mykiss). In this study, we identified 39 htr genes, including 14 htr1, 4 htr2, 4 htr2 like, 3 htr3, 4 htr4, 2 htr5, 2 htr6, and 6 htr7 subtypes. We investigated physiological functions of serotonin receptors in response to bacterial pathogens exposure and salinity changes. We showed htr1, htr2, htr4 and htr7 subtypes were associated with immunomodulation in response to Vibrio anguillarum or Aeromonas salmonicida infection. Saltwater (salinity of 15) transfer significantly altered htr1, htr2, htr4, and htr7 subtypes, suggesting trout Htr was associated with osmoregulation. We further showed residues interacted with inverse agonist (methiothepin) and serotonin analogue (5-Carboxamidotryptamine) were conserved between trout and human, suggesting exogenous ligands targeting human HTRs might have a role in aquaculture. This study showed duplicated trout Htrs might be physiologically neofunctionalized and potentially exhibit pleiotropic effects in regulating immunomodulation and osmoregulation.
Topics: Animals; Humans; Oncorhynchus mykiss; Serotonin; Drug Inverse Agonism; Salinity; Receptors, Serotonin; Bacterial Infections
PubMed: 37481174
DOI: 10.1016/j.ijbiomac.2023.125930 -
Virus Research Sep 2023Aeromonas salmonicida subsp. salmonicida is a Gam-negative bacterium responsible for furunculosis in fish. Because this aquatic bacterial pathogen has a rich reservoir...
Aeromonas salmonicida subsp. salmonicida is a Gam-negative bacterium responsible for furunculosis in fish. Because this aquatic bacterial pathogen has a rich reservoir of antibiotic-resistant genes, it is essential to investigate antibacterial alternatives, including the use of phages. Yet, we have previously demonstrated the inefficiency of a phage cocktail designed against A. salmonicida subsp. salmonicida strains due to a phage resistance phenotype associated to a prophage, namely Prophage 3. To bypass this resistance, one of the solutions is to isolate novel phages capable of infecting Prophage 3-bearing strains. Here we report on the isolation and characterization of the new virulent phage vB_AsaP_MQM1 (or MQM1), which is highly specific to A. salmonicida subsp. salmonicida strains. Phage MQM1 inhibited the growth of 01-B516, a strain carrying Prophage 3, including when combined to the previous phage cocktail. MQM1 infected 26 out of the 30 (87%) Prophage 3-bearing strains tested. Its linear dsDNA genome contains 63,343 bp, with a GC content of 50.2%. MQM1 genome can encode 88 proteins and 8 tRNAs, while no integrase or transposase-encoding genes were found. This podophage has an icosahedral capsid and a non-contractile short tail. We suggest that MQM1 may be a good addition to future phage cocktails against furunculosis to resolve the Prophage 3-resistance issue.
Topics: Animals; Bacteriophages; Furunculosis; Prophages; Aeromonas salmonicida; Fishes
PubMed: 37385348
DOI: 10.1016/j.virusres.2023.199165 -
RNA (New York, N.Y.) Sep 2023Unique chemical and physical properties are introduced by inserting selenocysteine (Sec) at specific sites within proteins. Recombinant and facile production of...
Unique chemical and physical properties are introduced by inserting selenocysteine (Sec) at specific sites within proteins. Recombinant and facile production of eukaryotic selenoproteins would benefit from a yeast expression system; however, the selenoprotein biosynthetic pathway was lost in the evolution of the kingdom Fungi as it diverged from its eukaryotic relatives. Based on our previous development of efficient selenoprotein production in bacteria, we designed a novel Sec biosynthesis pathway in using translation components. tRNA was mutated to resemble tRNA to allow recognition by seryl-tRNA synthetase as well as selenocysteine synthase (SelA) and selenophosphate synthetase (SelD). Expression of these Sec pathway components was then combined with metabolic engineering of yeast to enable the production of active methionine sulfate reductase enzyme containing genetically encoded Sec. Our report is the first demonstration that yeast is capable of selenoprotein production by site-specific incorporation of Sec.
Topics: Codon, Terminator; Saccharomyces cerevisiae; Aeromonas salmonicida; Protein Engineering; RNA, Transfer, Cys; Humans; Nucleic Acid Conformation
PubMed: 37279998
DOI: 10.1261/rna.079658.123 -
Fish & Shellfish Immunology Aug 2023The Yellow River carp (Cyprinus carpio haematopterus) is a vital economically farmed fish of the Cyprinidae family. With the development of intensive aquaculture, carp...
The Yellow River carp (Cyprinus carpio haematopterus) is a vital economically farmed fish of the Cyprinidae family. With the development of intensive aquaculture, carp production has increased dramatically, leading to the frequent occurrence of various diseases. Cell lines are considered the most cost-effective resource for in vitro studies and are widely used for physiological and pathological studies because of accessibility and convenience. This research established a novel immortal cell line CCM (Yellow River carp muscle cells) derived from the carp muscle. CCM has been passed over 71 generations for 1 year. The morphology of CCM and the adhesion and extension processes were captured by light and electron microscopy. CCM were passaged every 3 days with 20% FBS DMEM/F12 at 1:3. The optimum conditions for CCM growth were 28 °C and 20% FBS concentration. DNA sequencing of 16S rRNA and COI showed that CCM was derived from carp. CCM positively reacts to anti-PAX7 and anti-MyoD antibodies of carp. Analysis of chromosomes revealed that the chromosomal pattern number of CCM was 100. Transfection experiment demonstrated that CCM might be utilized to express foreign genes. Furthermore, cytotoxicity testing showed that CCM was susceptible to Aeromonas hydrophila, Aeromonas salmonicida, Aeromonas veronii, and Staphylococcus Aureus. The organophosphate pesticides (chlorpyrifos and glyphosate) or heavy metals (Hg, Cd, and Cu) exhibited dose-dependent cytotoxicity against CCM. After LPS treatment, the MyD88-IRAKs-NFκB pathway stimulates inflammatory-related factor il1β, il8, il10, and nfκb expression. LPS did not seem to cause oxidative stress in CCM, and the expression of cat and sod was not affected. Poly (I:C) through TLR3-TRIF-MyD88-TRAF6-NFκB and TRIF-TRAF3-TBK1-IRF3 activated the transcription of related factors, increased expression of anti-viral protein, but no changes in apoptosis-related genes. To our knowledge, this is the first muscle cell line in Yellow River carp and the first study on the immune response signal pathways of Yellow River carp based on the muscle cell line. CCM cell line provides a more rapid and efficient experimental material for fish immunology research, and this study preliminarily elucidated its immune response strategy to LPS and poly (I:C).
Topics: Animals; Carps; RNA, Ribosomal, 16S; Lipopolysaccharides; Myeloid Differentiation Factor 88; Poly I-C; Muscles; Muscle Cells; Cell Line; Adaptor Proteins, Vesicular Transport; Fish Diseases
PubMed: 37277052
DOI: 10.1016/j.fsi.2023.108859 -
Fish & Shellfish Immunology Aug 2023In recent years, more than one pathogenic organism has usually been isolated from diseased turbot Scophthalmus maximus, creating a pressing need for the development of...
In recent years, more than one pathogenic organism has usually been isolated from diseased turbot Scophthalmus maximus, creating a pressing need for the development of combination vaccines to prevent fish diseases brought on simultaneously by various infections. In this study, the inactivated bivalent vaccine of Aeromonas salmonicida and Edwardsiella tarda was prepared by the formalin inactivation method. After challenge with A. salmonicida and E. tarda at 4 weeks post-vaccination in turbot, the relative percentage survival (RPS) of the inactivated bivalent vaccine was 77.1%. In addition, we assessed the effects of the inactivated bivalent vaccine and evaluated the immunological processes after immunization in a turbot model. Serum antibody titer and lysozyme activity of the vaccinated group were both upregulated and higher than that in control group after vaccination. The expression levels of genes (TLR2, IL-1β, CD4, MHCI, MHCⅡ) that related to antigen recognition, processing and presentation were also studied in the liver, spleen and kidney tissues of vaccinated turbot. All the detected genes in the vaccinated group had a significant upward trend, and most of them reached the maximum value at 3-4 weeks, which had significant differences from the control group, suggesting that antigen recognition, processing and presentation pathway was activated by the inactivated bivalent vaccine. Our study provides a basis for further application of the killed bivalent vaccine against A. salmonicida and E. tarda in turbot, making it good potential that can be applied in aquaculture.
Topics: Animals; Enterobacteriaceae Infections; Edwardsiella tarda; Aeromonas salmonicida; Flatfishes; Antibodies, Bacterial; Vaccines, Combined; Fish Diseases; Bacterial Vaccines
PubMed: 37269913
DOI: 10.1016/j.fsi.2023.108837 -
Food Chemistry Oct 2023Microbial spoilage of meat products is a significant problem in the food industry. Aeromonas salmonicida is a significant microorganism responsible for spoilage in...
The interaction of an effector protein Hap secreted by Aeromonas salmonicida and myofibrillar protein of meat: Possible mechanisms from structural changes to sites of molecular docking.
Microbial spoilage of meat products is a significant problem in the food industry. Aeromonas salmonicida is a significant microorganism responsible for spoilage in chilled meat. Its effector protein, hemagglutinin protease (Hap), has been identified as an effective substance for degrading meat proteins. The ability of Hap to hydrolyze myofibrillar proteins (MPs) in vitro demonstrated that Hap has obvious proteolytic activity, which could alter MPs' tertiary structure, secondary structure, and sulfhydryl groups. Moreover, Hap could significantly degrade MPs, focusing primarily on myosin heavy chain (MHC) and actin. Active site analysis and molecular docking revealed that the active center of Hap was bound to MPs via hydrophobic interaction and hydrogen bonding. It may preferentially cleave peptide bonds between Gly44-Val45 in actin, and Ala825-Phe826 in MHC. These findings suggest that Hap may be involved in the spoilage mechanism of microorganisms and provide crucial insights into the mechanisms of meat spoilage induced by bacteria.
Topics: Aeromonas salmonicida; Molecular Docking Simulation; Actins; Meat; Proteolysis; Myosin Heavy Chains
PubMed: 37207606
DOI: 10.1016/j.foodchem.2023.136365 -
Applied Biochemistry and Biotechnology Feb 2024Aloe barbadensis Miller (ABM) is a traditional medicinal plant all over the world. Numerous studies were conducted to exhibit its medicinal properties and most of them...
Bioactive Analysis of Antibacterial Efficacy and Antioxidant Potential of Aloe barbadensis Miller Leaf Extracts and Exploration of Secondary Metabolites Using GC-MS Profiling.
Aloe barbadensis Miller (ABM) is a traditional medicinal plant all over the world. Numerous studies were conducted to exhibit its medicinal properties and most of them were concentrated on its metabolites against human pathogens. The current research work evaluates the attributes of different polar-based extracts (ethanol, methanol, ethyl acetate, acetone, hexane, and petroleum ether) of dried Aloe barbadensis leaf (ABL) to investigate its phytochemical constituents, antioxidant potential (DPPH, ABTS), phenolic, tannin, flavonoid contents, identification of bioactive compounds, and functional groups by gas chromatography-mass spectrometry (GC-MS) and fourier transform infrared spectroscopy (FT-IR) respectively, and comparing antibacterial efficacy against human pathogens, aquatic bacterial pathogens, and zoonotic bacteria associated with fish and human. The present results showed that the methanolic extract of ABL showed higher antioxidant activity (DPPH-59.73 ± 2.01%; ABTS-74.1 ± 1.29%), total phenolic (10.660 ± 1.242 mg GAE/g), tannin (7.158 ± 0.668 mg TAE/g), and flavonoid content (49.545 ± 1.928 µg QE/g) than that of other solvent extracts. Non-polar solvents hexane and petroleum ether exhibited lesser activity among the extracts. In the case of antibacterial activity, higher inhibition zone was recorded in methanol extract of ABL (25.00 ± 0.70 mm) against Aeromonas salmonicida. Variations in antibacterial activity were observed depending on solvents and extracts. In the current study, polar solvents revealed higher antibacterial activity when compared to the non-polar and the mid-polar solvents. Diverse crucial bioactive compounds were detected in GC-MS analysis. The vital compounds were hexadecanoic acid (30.69%) and 2-pentanone, 4-hydroxy-4-methyl (23.77%) which are responsible for higher antioxidant and antibacterial activity. Similar functional groups were identified in all the solvent extracts of ABL with slight variations in the FT-IR analysis. Polar-based solvent extraction influenced the elution of phytocompounds more than that of the other solvents used in this study. The obtained results suggested that the ABM could be an excellent source for antioxidant and antibacterial activities and can also serve as a potential source of effective bioactive compounds to combat human as well as aquatic pathogens.
Topics: Animals; Humans; Antioxidants; Tannins; Plant Extracts; Gas Chromatography-Mass Spectrometry; Aloe; Hexanes; Methanol; Spectroscopy, Fourier Transform Infrared; Anti-Bacterial Agents; Solvents; Phytochemicals; Flavonoids; Alkanes; Sulfonic Acids; Benzothiazoles
PubMed: 37184725
DOI: 10.1007/s12010-023-04565-z