bassianolide and Bacterial-Infections

Diseases of striped bass, their hybrids, and redfish (red drum) are important constraints to the culture of these two species. Since striped bass have been cultured for years the organisms that cause most diseases of these fish are well known, but very little specific disease information exists for redfish. However, it appears that the organisms that cause diseases of striped bass and redfish do not differ greatly from those of other fishes. The most significant viral disease is lymphocystis, but infectious pancreatic necrosis has occurred in striped bass. Vibriosis (Vibrio sp.) and motile Aeromonas septicemia (Aeromonas hydrophila) are the most frequently encountered bacterial diseases. Both species of fish are affected by fungi (usually Saprolegnia) when the fish are injured or stressed. Amyloodinium ocellatum is the most serious protozoan that infects striped bass and redfish, but the other common protozoans (Trichodina, Ichthyophthirius, Cryptocaron, etc.) have also been reported. Treatment of any of these diseases is a problem because of the absence of approved drugs or chemicals for use on striped bass or redfish. The most common therapeutics used on striped bass and redfish are copper sulfate, formalin, salt (in freshwater) and Terramycin.

Topics: Animals; Anti-Infective Agents; Bacterial Infections; Bass; Fish Diseases; Fishes; Mycoses; Parasitic Diseases; Parasitic Diseases, Animal; Virus Diseases

Fc receptors (FcRs) are key players in antibody-dependent cellular phagocytosis (ADCP) with their specific recognition of the Fc portion of an immunoglobulin. Despite reports of FcγR-mediated phagocytosis in mammals, little is known about the effects of soluble FcγRs on the immune response. In this study, FcγRIα was cloned from the largemouth bass (

Topics: Animals; Bacterial Infections; Bass; HEK293 Cells; Humans; Mammals; Receptors, IgG

Azomite is a hydrated calcium sodium aluminosilicat rich in rare earth elements. To investigate the dietary effects of Azomite on growth, intestine microbiota and morphology, immunohematological changes and disease resistance, seven diets with Azomite supplementation of 0 (the control), 1.0, 2.0, 3.0, 4.0, 5.0 and 6.0 g/kg (A0, A1, A2, A3, A4, A5, A6), were prepared and fed to largemouth bass, Micropterus salmoides (7.96 ± 0.19) for 60 days. The results revealed that the weight gain (WG) increased first and then decreased with the increasing dietary Azomite, and the A2 group presented the highest WG and lowest feed conversion ratio among all the groups. The supplementation of 2.0 g/kg Azomite significantly increased the intestine protease activity, the crude protein of whole body and protein retention (P < 0.05), and high inclusion of Azomite (6.0 g/kg) significantly reduced the lipid retention (P < 0.05). The amounts of red blood cells in A5, A6 groups, white blood cells in A3, A5, A6 groups and lymphocyte in A2-A6 groups were all significantly higher than those in the control group (P < 0.05). In addition, serum superoxide dismutase and catalase activities in A5, A6 groups, and serum alkaline phosphatase and lysozyme activities in A2-A4 groups showed significantly higher values than the control group (P < 0.05). Intestinal microbiota analysis indicated that the Tenericutes abundance was increased, whereas Proteobacteria abundance was decreased in all Azomite supplemented groups. The villus height in A2-A4 groups, and the villus width in A2 group were significantly higher than those of the control group (P < 0.05). The cumulative mortality was reduced by the addition of 2.0-5.0 g/kg Azomite after challenging with A. hydrophila (P < 0.05). In conclusion, proper addition of Azomite in diets improved the growth, intestine morphology, immune response and disease resistance in largemouth bass, and the optimal inclusion was estimated to be 2.0-3.0 g/kg diet.

Topics: Aluminum Silicates; Animal Feed; Animals; Bacterial Infections; Bass; Diet; Dietary Supplements; Disease Resistance; Dose-Response Relationship, Drug; Fish Diseases; Gastrointestinal Microbiome; Random Allocation; Trace Elements

Aquaculture is responsible for more than 50% of global seafood consumption. Bacterial diseases are a major constraint to this sector and associated with misuse of antibiotics, pose serious threats to public health. Fish-symbionts, co-inhabitants of fish pathogens, might be a promising source of natural antimicrobial compounds (NACs) alternative to antibiotics, limiting bacterial diseases occurrence in aquafarms. In particular, sporeforming Bacillus spp. are known for their probiotic potential and production of NACs antagonistic of bacterial pathogens and are abundant in aquaculture fish guts. Harnessing the fish-gut microbial community potential, 172 sporeforming strains producing NACs were isolated from economically important aquaculture fish species, namely European seabass, gilthead seabream, and white seabream. We demonstrated that they possess anti-growth, anti-biofilm, or anti-quorum-sensing activities, to control bacterial infections and 52% of these isolates effectively antagonized important fish pathogens, including Aeromonas hydrophila, A. salmonicida, A. bivalvium, A. veronii, Vibrio anguillarum, V. harveyi, V. parahaemolyticus, V. vulnificus, Photobacterium damselae, Tenacibaculum maritimum, Edwardsiela tarda, and Shigella sonnei. By in vitro quantification of sporeformers' capacity to suppress growth and biofilm formation of fish pathogens, and by assessing their potential to interfere with pathogens communication, we identified three promising candidates to become probiotics or source of bioactive molecules to be used in aquaculture against bacterial aquaculture diseases.

Topics: Animals; Anti-Bacterial Agents; Aquaculture; Bacillus; Bacterial Infections; Bass; Biofilms; Fish Diseases; Gastrointestinal Microbiome; Probiotics; Quorum Sensing; Sea Bream

Mannose-binding lectin (MBL) plays an important role in host immune responses against pathogens. LjMBL-like-1 was identified from Japanese sea bass (Lateolabrax japonicas), which has selectivity for galactose. Herein, this lectin might be better designated as galactose-binding lectin (LjGalBL-1). LjGalBL-1 transcripts were detected in all tested tissues, with highest expression in liver. Upon Vibrio harveyi infection, LjGalBL-1 mRNA expression was increased in major immune-related tissues, and protein levels in serum were also upregulated. Recombinant LjGalBL-1 (rLjGalBL-1) bound to monosaccharides and polysaccharides, and both rLjGalBL-1 and native LjGalBL-1 (nLjGalBL-1) agglutinated three Gram-positive bacteria (Staphylococcus aureus, Streptococcus iniae and Micrococcus luteus) and four Gram-negative bacteria (Aeromonas hydrophila, Edwardsiella tarda, Vibrio anguillarum and V. harveyi) in a Ca

Topics: Agglutination; Animals; Bacterial Infections; Bass; Calcium Signaling; Cloning, Molecular; Fish Proteins; Immunity, Innate; Liver; Mannose-Binding Lectins; Sequence Alignment; Transcriptome; Up-Regulation

Bacterial fish diseases constitute a major problem in aquaculture, it was found in the environment and under stressors cause severe economic losses to fish. This work aimed to investigate the bacterial causes and suitable treatments of mass mortality in some cultured marine fish farms in Damietta governorate.. The study was performed on 5 farms suffered from mass mortality. Total of 100 diseased fish (10 sea bass and 10 sea bream/farm) and 20 water samples were randomly collected from these farms. Bacteriological examinations were carried out followed by in vitro sensitivity tests. Treatment trial was performed using the most effective antibacterial agent on isolated bacteria.. From fish and water samples Pseudomonas spp., Aeromonas spp. and Vibrio spp. were isolated with the rat of (16, 10%), (22, 10%) and (28, 10%) respectively. These results were confirmed biochemically. Some virulence genes of isolated bacteria were detected using PCR; meanwhile, enrofloxacin reduced significantly the mortality rates in examined farms.. It could be concluded that, Pseudomonas spp., Aeromonas spp. and Vibrio spp. are the main bacterial species causing mass mortality in marine fish farms. These bacteria were highly sensitive to enrofloxacin in vitro and in vivo.

Topics: Animals; Anti-Bacterial Agents; Bacteria; Bacterial Infections; Bass; Enrofloxacin; Fish Diseases; Fisheries; Microbial Sensitivity Tests; Polymerase Chain Reaction; Sea Bream; Virulence

A thorough understanding of host-microbe interactions is crucial for more efficient disease management in the marine larviculture industry. As demonstrated in terrestrial animal research, gnotobiotic systems (involving animals cultured in germ-free conditions or inoculated with known microorganisms) are excellent tools to extend our understanding of the mechanisms involved in host-microbe interactions and allow the evaluation of new treatments for diseases. In this study, we introduce a germ-free European sea bass Dicentrarchus labrax larval model, independent of the continuous addition of antimicrobial agents. This model has an experimental set-up that allows addition of live feed to the larvae without compromising the germ-free status. This model will facilitate and render aquaculture research more effective in terms of mitigation fish larval diseases.

Topics: Animals; Bacterial Infections; Bass; Disinfectants; Fish Diseases; Germ-Free Life; Larva; Ovum

The discovery of rhamnose-binding lectins (RBLs) in teleost fish eggs led to the identification of a novel lectin family characterized by a unique sequence motif and a structural fold, and initially proposed to modulate fertilization. Further studies of the RBL tissue localization and gene organization were also suggestive of role(s) in innate immunity. Here we describe the purification, and biochemical and functional characterization of a novel RBL (DlRBL) from sea bass (Dicentrarchus labrax) serum. The purified DlRBL had electrophoretic mobilities corresponding to 24 kDa and 100 kDa under reducing and non-reducing conditions, respectively, suggesting that in plasma the DlRBL is present as a physiological homotetramer. DlRBL subunit transcripts revealed an open reading frame encoding 212 amino acid residues that included two tandemly-arrayed carbohydrate-recognition domains, and an 18-residue signal sequence at the N-terminus. The deduced size of 24.1 kDa for the mature protein was in good agreement with the subunit size of the isolated lectin. Binding activity of DlRBL for rabbit erythrocytes could be inhibited in the presence of rhamnose or galactose, did not require calcium, and was optimal at around 20°C and within the pH 6.5-8.0 range. DlRBL agglutinated Gram positive and Gram negative bacteria, and exposure of formalin-killed Escherichia coli to DlRBL enhanced their phagocytosis by D. labrax peritoneal macrophages relative to the unexposed controls. Taken together, the results suggest that plasma DlRBL may play a role in immune recognition of microbial pathogens and facilitate their clearance by phagocytosis.

Topics: Agglutination; Amino Acid Motifs; Amino Acid Sequence; Animals; Bacterial Infections; Bass; Erythrocytes; Escherichia coli; Immunity, Innate; Lectins; Macrophages, Peritoneal; Molecular Sequence Data; Phagocytosis; Phylogeny; Plasma; Protein Binding; Protein Multimerization; Protein Structure, Tertiary; Rabbits; Rhamnose

The orange-spotted grouper (Epinephelus coioides), a favorite marine food fish, is widely cultured in China and Southeast Asian countries. However, little is known about its acute phase response (APR) caused by viral diseases. Serum amyloid A (SAA) is a major acute phase protein (APP). In this study, a new SAA homologous (EcSAA) gene was cloned from grouper, E. coioides, by rapid amplification of cDNA ends (RACE) PCR. The full-length cDNA sequence of SAA was 508 bp and contained a 363 bp open reading frame (ORF) coding for a protein of 121 aa. Similar to other fish known SAA genes, the EcSAA gene contained four exons and three introns. Quantitative real-time PCR analysis revealed that EcSAA mRNA is predominately expressed in liver and gill of grouper. Furthermore, the expression of EcSAA was differentially up-regulated in liver after infection with Staphyloccocus aureus, Vibrio vulnificus, Vibrio parahaemolyticus, Saccharomyces cerevisiae and Singapore grouper iridovirus (SGIV). Recombinant EcSAA (rEcSAA) was expressed in Escherichia BL21 (DE3) and purified for mouse anti-EcSAA serum preparation. The rEcSAA fusion protein was demonstrated to bind to all tested bacteria and yeast, and inhibit the replication of SGIV. Overexpression of EcSAA in grouper spleen (GS) cells could also inhibit the replication of SGIV. These results suggest that EcSAA may be an important molecule in the innate immunity of grouper.

Topics: Amino Acid Sequence; Animals; Bacteria; Bacterial Infections; Bass; Cloning, Molecular; DNA Virus Infections; DNA, Complementary; Fish Diseases; Fish Proteins; Gene Expression Regulation; Immunity, Innate; Molecular Sequence Data; Organ Specificity; Phylogeny; Polymerase Chain Reaction; Ranavirus; RNA, Messenger; Saccharomyces cerevisiae; Sequence Alignment; Sequence Analysis, DNA; Serum Amyloid A Protein

Orange-spotted grouper, Epinephelus coioides is one of the most important economic species of marine-cultured fish in China and Southeast Asia countries. However, very little information of the innate immune mechanisms against microbial pathogens is available in grouper, Epinephelus sp. Hepcidin, as an antimicrobial peptide (AMP), is a very important component in the innate immune system and widespread in fish. In this study, two novel types of hepcidin gene (designated EC-hepcidin1 and EC-hepcidin2) were cloned from E. coioides. They consist of open reading frames (ORFs) of 267 bp and 263 bp encoding the putative peptides of 88 and 87 amino acids, respectively. The homologous identity of deduced amino acid sequences between EC-hepcidin1 and EC-hepcidin2 is up to 79%, and predicted mature regions of both them have four cysteines residues. Genomic DNAs of both EC-hepcidin1 and EC-hepcidin2 consist of three exons and two introns. RT-PCR results showed that EC-hepcidin1 transcript was most abundant in liver and less in stomach. However, the transcript of EC-hepcidin2 was only detected in liver. The expressions of both EC-hepcidins were up-regulated by microbial and viral challenges, and iron overload, respectively, and EC-hepcidin1 was more responsive. The growth of Gram-negative bacterium of Vibrio vulnificus and Gram-positive bacterium of Staphylococcus aureus was inhibited by synthetic EC-hepcidins, and EC-hepcidin1 displayed stronger antimicrobial activity. The replication of Singapore grouper iridovirus (SGIV) was inhibited in the EC-hepcidin1 and EC-hepcidin2 over-expressed stable transfected fish cell lines (GS/pcDNA-Hep1, GS/pcDNA-Hep2) indicative of the antiviral activity of EC-hepcidins. These data should offer important information on the antimicrobial and antiviral roles of EC-hepcidins, and will be help to the better understanding of molecular mechanisms of grouper innate immunity.

Topics: Amino Acid Sequence; Animals; Anti-Bacterial Agents; Antimicrobial Cationic Peptides; Antiviral Agents; Bacteria; Bacterial Infections; Base Sequence; Bass; Cloning, Molecular; DNA Virus Infections; Gene Expression Profiling; Gene Expression Regulation; Gene Order; Hepcidins; Iridovirus; Iron; Molecular Sequence Data; RNA, Messenger; Saccharomyces cerevisiae; Sequence Alignment

Triggering of phagocyte apoptosis is a major virulence mechanism used by some successful bacterial pathogens. A central issue in the apoptotic death context is that fully developed apoptosis results in necrotic cell autolysis (secondary necrosis) with release of harmful cell components. In multicellular animals, this occurs when apoptosing cells are not removed by scavengers, mainly macrophages. Secondary necrotic lysis of neutrophils and macrophages may occur in infection when extensive phagocyte apoptosis is induced by bacterial cytotoxins and removal of apoptosing phagocytes is defective because the apoptotic process exceeds the available scavenging capacity or targets macrophages directly. Induction of phagocyte secondary necrosis is an important pathogenic mechanism, as it combines the pathogen evasion from phagocyte antimicrobial activities and the release of highly cytotoxic molecules, particularly of neutrophil origin, such as neutrophil elastase. This pathogenicity mechanism therefore promotes the unrestricted multiplication of the pathogen and contributes directly to the pathology of several necrotizing infections, where extensive apoptosis and necrosis of macrophages and neutrophils are present. Here, examples of necrotizing infectious diseases, where phagocyte secondary necrosis is implicated, are reviewed.

Topics: Animals; Apoptosis; Autolysis; Bacterial Infections; Bass; Necrosis; Phagocytes; Phagocytosis; Signal Transduction; Virulence

The use of probiotic microorganisms in aquaculture is gaining a lot of interest. Gnotobiotic model systems are required in order to fully understand the effects and modes-of-action of these microorganisms, as the native microbial communities present in non-sterile animals can lead to false conclusions. In this study, a gnotobiotic sea bass larvae (Dicentrarchus labrax) test system was developed. In order to obtain bacteria-free animals, the eggs were disinfected with glutaraldehyde and subsequently incubated in a solution of rifampicin and ampicillin. Axenity was confirmed using culture-dependent and -independent techniques. The gnotobiotic larvae were fed axenic Artemia sp. from 7 days after hatching onwards. In the challenge test, one of the three opportunistic pathogens, Aeromonas hydrophila, Listonella anguillarum serovar O1 and O2a, was added to the model system via the water and encapsulated in Artemia sp. Only serovar O2a led to increased mortality in the sea bass larvae. The presented gnotobiotic model can be used for research on, among others, reciprocal metabolic effects between microorganisms and the host (e.g. as measured by gene expression), immunostimulants, pharmacological research and the histological development of the gastrointestinal tract and growth of larvae.

Topics: Aeromonas hydrophila; Animals; Bacterial Infections; Bass; Colony Count, Microbial; Disease Models, Animal; Germ-Free Life; Larva; Listonella; Survival Analysis

Epinecidin-1 is an antimicrobial peptide and plays a vital role in protecting fish against pathogenic infection. As a mimic of a grouper epinecidin-1 peptide, it has tertiary structures that closely resemble those of pleurocidin found in the winter flounder (Pleuronectes americanus). The tissue-specific, lipopolysaccharide (LPS)-stimulation-specific, and poly(I):poly(C)-stimulation-specific expressions of the grouper (Epinephelus coioides) epinecidin-1 antimicrobial peptide were determined using a comparative reverse-transcription polymerase chain reaction. Results of the tissue distribution analysis revealed high levels of epinecidin-1 messenger RNA (mRNA) in the head kidneys, intestines, and skin. Expression of epinecidin-1 mRNA was dose-dependently stimulated by both LPS and poly(I):poly(C). Immunohistochemical analysis with the polyclonal antiserum of a grouper epinecidin-1 peptide (rabbit polyclonal antibody) showed that the peptide was localized with the epinecidin-1 antibody in the gills and intestines. Two synthetic peptides of the grouper epinecidin-1 peptide (g-ple 22-51 and g-ple 22-42) and one winter flounder pleurocidin as a control exhibited high antimicrobial activities against gram-negative or gram-positive bacteria. In addition, peptide treatment was effective in promoting a significant increase in fish survival after the injection of Vibrio vulnificus in tilapia (Oreochromis mossambicus) and grouper. These results are relevant to the design of prophylactic and therapeutic strategies to counter bacterial infections, especially for preventing or ameliorating immune defects in fish during bacterial infections.

Topics: Amino Acid Sequence; Animals; Antimicrobial Cationic Peptides; Bacterial Infections; Base Sequence; Bass; DNA Primers; Fish Diseases; Fish Proteins; Gene Expression; Gills; Intestinal Mucosa; Models, Molecular; Molecular Sequence Data; Recombinant Proteins; RNA, Messenger

A hepcidin gene was amplified from the liver of Lateolabrax japonicus challenged with a mixed bacterial suspension. Using RT-PCR and RACE, a full length cDNA sequence of the hepcidin like antimicrobial peptide was determined. The complete hepcidin cDNA Hepc2 is 581 bases and contains an ORF of 258 bases with a coding capacity of 86 amino acids. The deduced amino acid sequence, which shares eight cysteines at the identical conserved positions, and gene organisation are conserved between Japan sea bass and other fish species. The predicted molecular weight of the peptide is 9.4 kDa. The 3'-untranslated region is composed of 225 bp with a polyadenylation signal AATAAA sequence appearing at 189 nt and the poly(A) tail at 212 nt downstream of stop codon TGA. The predicted signal peptide cleavage site of its deduced peptide is between codons 24 and 25. Japan sea bass hepcidin-like genomic DNA hepc2 sequence including upstream and downstream regions was composed of two introns and three exons. The cloned 173-bp upstream sequence of Japan sea bass hepcidin-like gene contains putative regulatory elements and several binding motifs for transcription factors. High homologies with hepcidin cDNAs and peptides of white bass (Morone chrysops), human and other fish were shown. Hepc2 of Lateolabrax japonicus is a new member of the hepcidin gene family.

Topics: Amino Acid Sequence; Animals; Antimicrobial Cationic Peptides; Bacterial Infections; Base Sequence; Bass; Cloning, Molecular; DNA Primers; DNA, Complementary; Fish Diseases; Hepcidins; Humans; Liver; Molecular Sequence Data; Phylogeny; Reverse Transcriptase Polymerase Chain Reaction; Sequence Alignment; Sequence Homology, Amino Acid

Bass hepcidin was purified from the gill of hybrid striped bass (Morone chrysops x Morone saxatilis) based on antimicrobial activity against Escherichia coli. This 21-amino acid peptide has 8 cysteines engaged in 4 disulfide bonds and is very similar to human hepcidin, an antimicrobial peptide with iron regulatory properties. To gain insight into potential role(s) of bass hepcidin in innate immunity in fish, we synthesized the peptide, characterized its antimicrobial activities in vitro, determined its solution structure by NMR, and quantified hepatic gene expression in vivo following infection of bass with the fish pathogens, Streptococcus iniae or Aeromonas salmonicida. Its structure is very similar to that of human hepcidin, including the presence of an antiparallel beta-sheet, a conserved disulfide-bonding pattern, and a rare vicinal disulfide bond. Synthetic bass hepcidin was active in vitro against Gram-negative pathogens and fungi but showed no activity against key Gram-positive pathogens and a single yeast strain tested. Hepcidin was non-hemolytic at microbicidal concentrations and had lower specific activity than moronecidin, a broad spectrum, amphipathic, alpha-helical, antimicrobial peptide constitutively expressed in bass gill tissue. Good synergism between the bacterial killing activities of hepcidin and moronecidin was observed in vitro. Hepcidin gene expression in bass liver increased significantly within hours of infection with Gram-positive (S. iniae) or Gram-negative (A. salmonicida) pathogens and was 4-5 orders of magnitude above base-line 24-48 h post-infection. Our results suggest that hepcidin plays a key role in the antimicrobial defenses of bass and that its functions are potentially conserved between fish and human.

Topics: Animals; Antimicrobial Cationic Peptides; Bacterial Infections; Base Sequence; Bass; Chromatography, High Pressure Liquid; Disulfides; DNA Primers; Fish Diseases; Hepcidins; Liver; Magnetic Resonance Spectroscopy; Models, Molecular; Molecular Sequence Data; Protein Conformation; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Topics: Aeromonas; Animals; Aquaculture; Bacterial Infections; Bass; Fish Diseases; Histological Techniques; Hybridization, Genetic; Liver; Mycobacterium; Vibrio