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Fish & Shellfish Immunology Jun 2024Meningitis caused by Gram-negative bacteria is a serious public health problem, causing morbidity and mortality in both children and adults. Here, we propose a novel...
Meningitis caused by Gram-negative bacteria is a serious public health problem, causing morbidity and mortality in both children and adults. Here, we propose a novel experimental model using Nile tilapia (Oreochromis niloticus) to study neuroinflammation. The fish were infected with Aeromonas hydrophila, and the course of infection was monitored in the peripheral blood. Septicemia was obvious in the blood, while in the brain tissue, infection of the meninges was present. The histopathological examination showed suppurative meningitis, and the cellular immune response in the brain tissue during infection was mediated by microglia. These cells were morphologically characterized and phenotyped by MHC class II markers and CD68. The increased production of TNF-α, IL-1β and iNOS supported the infiltration of these cells during the neuroinflammatory process. In the proteomic analysis of A. hydrophila isolated from brain tissue, we found chemotactic and transport proteins, proteolytic enzymes and enzymes associated with the dismutation of nitric oxide (NO), as well as motor proteins and those responsible for cell division. After characterizing the most abundant proteins during the course of infection, we investigated the druggability index of these proteins and identified promising peptide sequences as molecular targets that are similar among bacteria. Thus, these findings deepened the understanding of the pathophysiology of meningitis caused by A. hydrophila. Moreover, through the proteomics analysis, important mechanisms and pathways used by the pathogen to subvert the host response were revealed, providing insights for the development of novel antibiotics and vaccines.
PubMed: 38866348
DOI: 10.1016/j.fsi.2024.109687 -
Scientific Reports Jun 2024In this study, 80 catfish fillets were randomly collected from Egyptian local markets and retailers. The samples included 20 African catfish (Clarias gariepinus), 20...
In this study, 80 catfish fillets were randomly collected from Egyptian local markets and retailers. The samples included 20 African catfish (Clarias gariepinus), 20 bayad (Bagrus bajad), and 40 pangasius catfish (Pangasianodon hypophthalmus) fillets. Pangasianodon hypophthalmus fillet samples were divided into 20 white basa and 20 red basa fillets. We conducted a microbiological analysis of catfish fillet samples, evaluating mesophilic aerobic bacteria, psychrophilic aerobic bacteria, HS-producing bacteria, Staphylococcus spp., Enterobacteriaceae, Coliforms, and fecal Coliform counts. Additionally, we identified the existence of Salmonella spp., Vibrio spp., Yersinia spp., Escherichia spp., Aeromonas spp., and Pseudomonas spp. in the catfish fillet samples. In our study, the psychrophilic bacterial counts in Bagrus bajad (5.21 log CFU/g) were found to be higher compared to the counts in Clarias gariepinus (4.31 log CFU/g) and Pangasianodon hypophthalmus (3.89-4.7 log CFU/g). The fecal Coliform in Clarias gariepinus fillets was significantly higher than in other catfish fillets. We isolated Escherichia coli, Escherichia fergusonii, Aeromonas hydrophila, and Pseudomonas luteola from the catfish fillets, while no Salmonella spp., Vibrio spp., or Yersinia spp. were detected. These isolates were identified using 16S rRNA sequencing and phylogenetic analysis. Furthermore, ten Escherichia spp. were serologically identified, revealing that O26 and O78 were the most commonly occurring serotypes. This study highlights the microbiological analysis conducted on catfish fillets and concludes that the fillet samples from these catfish were of superior quality and deemed acceptable for human consumption.
Topics: Animals; Catfishes; Food Microbiology; Bacteria; Seafood
PubMed: 38858383
DOI: 10.1038/s41598-024-62730-8 -
Journal of Animal Physiology and Animal... Jun 2024This experiment was performed to identify the influence of dietary Saussurea lappa root (SLR) on the performance and general health status of Nile Tilapia fingerlings...
The effect of dietary administration of Saussurea lappa root on performance, blood biochemical indices, redox status, innate immune response, intestinal microbial population and resistance against A. hydrophila infections of Tilapia Fingerlings.
This experiment was performed to identify the influence of dietary Saussurea lappa root (SLR) on the performance and general health status of Nile Tilapia fingerlings (O. niloticus). Four formulated diets with different SLR levels of 0.0, 2.5, 5 and 10 g/kg, respectively, were afforded to fingerling fish (15.42 ± 0.05 g) for 8 weeks. The feed efficiency ratio (FER), feed intake (FI) and feed conversion ratio varied with dietary SLR level in a linear model and a high feed efficiency rate was recorded at the 10 g/kg group, while FI and FCR exhibited an opposite trend (P < 0.001). Dietary SLR level influenced serum protein constituents, liver and renal function enzymes, triglycerides, cholesterol and glucose (P < 0.001). Serum Catalase (CAT), total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) remarkedly increased with dietary SLR level and attained a level at 10 g/kg. Furthermore, serum lysozyme, complement C3 (C3), IgA and IgD were stimulated by 10 g/kg SLR. Intestinal digestive enzymes (lipase and amylase) increased with SLR level up to 10 g/kg. As the dietary SLR level raised, the cumulative survival percentage aginst A. hydrophila challenge increased and then reached a maximum at 10 g/kg SLR group. Moreover, gene expression of pro-inflammation cytokines (TNF-2a, IL-1β, and IL-10) in liver and kidney transcriptomes demonstrated effective immunostimulant capabilities of greater SLR inclusion levels in fish diet. Meanwhile, intestinal microbial investigation, revealed that high levels of SLR in tilapia fish feed significantly suppressed total bacterial count, and pathogenic bacterial count (such as, E. coli, Coliform, Aeromonas spp, Pseudomonas spp.), and stimulated lactic acid bacteria development. Finally, it is recommended to include a high level of SLR (5 or 10 g/kg) in the diet of O. niloticus fingerlings to enhance feed efficiency, antioxidant characteristics, and immunological response against bacterial infections.
PubMed: 38851860
DOI: 10.1111/jpn.13998 -
Developmental and Comparative Immunology Sep 2024Interleukin (IL) 20 is a multifunctional cytokine and plays a vital role in regulating autoimmune diseases, inflammation, and immune responses. IL-20 homologs have been...
Interleukin (IL) 20 is a multifunctional cytokine and plays a vital role in regulating autoimmune diseases, inflammation, and immune responses. IL-20 homologs have been described in fish. However, due to the lack of antibodies, cellular sources and immunological functions of fish IL-20 in response to infections have not been fully characterized. In this study, a monoclonal antibody (mAb) was generated against the recombinant grass carp (Ctenopharyngodon idella) IL-20 protein and characterized by immunoblotting, immunofluorescent microscopy and flow cytometry. It was shown that the IL-20 mAb specifically recognized recombinant IL-20 proteins expressed in the E. coli cells and HEK293 cells. Using confocal microscopy, the IL-20 cells were identified in the head kidney, gills and intestine of grass carp, and induced after infection with Aeromonas hydrophila. Moreover, the IL-20 protein was found to be secreted mainly by CD3γδ T cells which were located predominantly in the gill filaments and intestinal mucosa. Taken together, our results suggest that IL-20 producing T cells are required for the mucosal immunity against bacterial infection in fish.
Topics: Animals; Carps; Aeromonas hydrophila; Gram-Negative Bacterial Infections; Fish Diseases; Fish Proteins; Humans; Immunity, Mucosal; Interleukins; HEK293 Cells; Gills; CD3 Complex; Antibodies, Monoclonal; Intestinal Mucosa; T-Lymphocytes; Mucous Membrane
PubMed: 38844187
DOI: 10.1016/j.dci.2024.105210 -
Ecotoxicology and Environmental Safety Jul 2024This study aimed to evaluate the effect of adding liquid extract of algae (Hypnea musciformis, Grateloupia acuminata, and Sargassum muticum) (HGS) and Magnesium oxide...
Synergistic effect of green synthesis magnesium oxide nanoparticles and seaweed extract on improving water quality, health benefits, and disease resistance in Nile tilapia.
This study aimed to evaluate the effect of adding liquid extract of algae (Hypnea musciformis, Grateloupia acuminata, and Sargassum muticum) (HGS) and Magnesium oxide nanoparticles (MgO NPs) using this extract to rear water of Oreochromis niloticus, on improving culture water indices, growth performance, digestive enzyme, hemato-biochemical characters, immune, antioxidative responses, and resistance after challenged by Aeromonas hydrophila with specific refer to the potential role of the mixture in vitro as resistance against three strains bacteria (Aeromonas sobria, Pseudomonas fluorescens, P. aeruginosa) and one parasite (Cichlidogyrus tilapia). The first group represented control, HGS0, whereas the other group, HGS5, HGS10, and HGS15 mL of liquid extract, as well as all groups with 7.5 μg mL MgO-NPs added to culture water of O. niloticus, for 60 days. Data showed that increasing levels at HGS 10 and HGS15 mL in to-culture water significantly enhanced growth-stimulating digestive enzyme activity and a significantly improved survival rate of O. niloticus after being challenged with A. hydrophila than in the control group. The total viability, coliform, fecal coliform count, and heavy metal in muscle partially decreased at HGS 10 and HGS15 mL than in the control group. Correspondingly, the highest positive effect on hemato-biochemical indices was noticed at levels HGS 10 and HGS15 mL. Fish noticed an improvement in immune and antioxidant indices compared to control groups partially at HGS 10 and HGS15 mL. Interestingly, fish cultured in rearing water with the mixture provided downregulated the related inflammatory genes (HSP70, TNF, IL-1β, and IL-8) partially at HGS15 mL. In vitro, the mixture showed positive efficiency as an antibacterial and partially antiparasitic at HGS 10 and HGS15 mL. This study proposes utilizing a mixture of (HGS) and (MgO-NPs) with optimum levels of 10-15 mL in cultured water to improve water indices, growth, health status, and increased resistance of O. niloticus against bacterial and parasitic infection.
Topics: Animals; Magnesium Oxide; Cichlids; Disease Resistance; Water Quality; Seaweed; Fish Diseases; Plant Extracts; Nanoparticles; Green Chemistry Technology; Metal Nanoparticles; Aeromonas hydrophila; Sargassum
PubMed: 38843743
DOI: 10.1016/j.ecoenv.2024.116522 -
BMC Veterinary Research Jun 2024The liver-expressed antimicrobial peptide 2 (LEAP2) plays a pivotal role in the host's immune response against pathogenic microorganisms. Numerous such antimicrobial...
BACKGROUND
The liver-expressed antimicrobial peptide 2 (LEAP2) plays a pivotal role in the host's immune response against pathogenic microorganisms. Numerous such antimicrobial peptides have recently been shown to mitigate infection risk in fish, and studying those harboured by the economically important fish Acrossocheilus fasciatus is imperative for enhancing its immune responses against pathogenic microorganisms. In this study, we cloned and sequenced LEAP2 cDNA from A. fasciatus to examine its expression in immune tissues and investigate the structure-activity relationships of its intramolecular disulphide bonds.
RESULTS
The predicted amino acid sequence of A. fasciatus LEAP2 was found to include a signal peptide, pro-domain, and mature peptide. Sequence analysis indicated that A. fasciatus LEAP2 is a member of the fish LEAP2A cluster and is closely related to Cyprinus carpio LEAP2A. A. fasciatus LEAP2 transcripts were expressed in various tissues, with the head kidney exhibiting the highest mRNA levels. Upon exposure to Aeromonas hydrophila infection, LEAP2 expression was significantly upregulated in the liver, head kidney, and spleen. A mature peptide of A. fasciatus LEAP2, consisting of two disulphide bonds (Af-LEAP2-cys), and a linear form of the LEAP2 mature peptide (Af-LEAP2) were chemically synthesised. The circular dichroism spectroscopy result shows differences between the secondary structures of Af-LEAP2 and Af-LEAP2-cys, with a lower proportion of alpha helix and a higher proportion of random coil in Af-LEAP2. Af-LEAP2 exhibited potent antimicrobial activity against most tested bacteria, including Acinetobacter guillouiae, Pseudomonas aeruginosa, Staphylococcus saprophyticus, and Staphylococcus warneri. In contrast, Af-LEAP2-cys demonstrated weak or no antibacterial activity against the tested bacteria. Af-LEAP2 had a disruptive effect on bacterial cell membrane integrity, whereas Af-LEAP2-cys did not exhibit this effect. Additionally, neither Af-LEAP2 nor Af-LEAP2-cys displayed any observable ability to hydrolyse the genomic DNA of P. aeruginosa.
CONCLUSIONS
Our study provides clear evidence that linear LEAP2 exhibits better antibacterial activity than oxidised LEAP2, thereby confirming, for the first time, this phenomenon in fish.
Topics: Animals; Structure-Activity Relationship; Amino Acid Sequence; Fish Diseases; Antimicrobial Peptides; Fish Proteins; Disulfides; Phylogeny; Aeromonas hydrophila; Base Sequence
PubMed: 38835040
DOI: 10.1186/s12917-024-04106-8 -
Letters in Applied Microbiology Jun 2024Although the genus Aeromonas inhabits the natural environment, it has also been isolated from hospital patient specimens as a causative agent of Aeromonas infections....
Rapid discrimination methods for clinical and environmental strains of Aeromonas hydrophila and A. veronii biovar sobria using the N-terminal sequence of the flaA gene and investigation of antimicrobial resistance.
Although the genus Aeromonas inhabits the natural environment, it has also been isolated from hospital patient specimens as a causative agent of Aeromonas infections. However, it is not known whether clinical strains live in the natural environment, and if these strains have acquired antimicrobial resistance. In this study, we performed the typing of flagellin A gene (flaA) of clinical and environmental strains of Aeromonas hydrophila and A. veronii biovar sobria using Polymerase Chain Reaction (PCR) assay with newly designed primers. Detection rates of the clinical and environmental flaA types of A. hydrophila were 66.7% and 88.2%, and the corresponding rates for A. veronii biovar sobria were 66.7% and 90.9%. The PCR assays could significantly discriminate between clinical and environmental strains of both species in approximately 4 h. Also, among the 63 clinical Aeromonas strains used, only one extended-spectrum β-lactamase-producing bacteria, no plasmid-mediated quinolone resistance bacteria, and only four multidrug-resistant bacteria were detected. Therefore, the PCR assays could be useful for the rapid diagnosis of these Aeromonas infections and the monitoring of clinical strain invasion into water-related facilities and environments. Also, the frequency of drug-resistant Aeromonas in clinical isolates from Okinawa Prefecture, Japan, appeared to be low.
Topics: Aeromonas hydrophila; Humans; Gram-Negative Bacterial Infections; Polymerase Chain Reaction; Flagellin; Aeromonas veronii; Drug Resistance, Bacterial; Anti-Bacterial Agents; Microbial Sensitivity Tests; Environmental Microbiology
PubMed: 38830808
DOI: 10.1093/lambio/ovae052 -
Fish & Shellfish Immunology Jun 2024Bacterial septicemia in freshwater fish is mainly caused by Aeromonas hydrophila infection, which affects the development of aquaculture industry. In the context of...
Bacterial septicemia in freshwater fish is mainly caused by Aeromonas hydrophila infection, which affects the development of aquaculture industry. In the context of sustainable aquaculture, subunit vaccines are of great values because they play positive roles in reducing the overuse of antibiotics and protecting aquatic animals against bacterial infection. In this study, the recombinant outer membrane protein OmpTS of A. hydrophila were used as subunit vaccine to immunize Megalobrama amblycephala, and its immunoprotective effect and host immune responses were evaluated. The survival rates of the vaccinated groups after bacterial infection were significantly higher than that of the control group, especially of the OmpTS high-dose vaccinated group. The better protective effects of vaccinated groups might be attributed to the increased levels of serum IgM-specific antibody titer, the reduced relative abundance of A. hydrophila in various tissues, the increased number of immune-positive cells with different epitopes, the up-regulated expression levels of immune-related genes, and the enhanced activities of antibacterial enzymes. In conclusion, OmpTS subunit vaccine could strongly induce host immune responses in M. amblycephala, thereby enhancing both cellular and humoral immunity, which exhibited excellent and effective immunoprotective efficacy.
PubMed: 38830521
DOI: 10.1016/j.fsi.2024.109665 -
F1000Research 2023Strategies to increase body resistance and prevent disease in aquaculture include using vaccines, antibiotics, and probiotics. Today, the use of antibiotics with...
Strategies to increase body resistance and prevent disease in aquaculture include using vaccines, antibiotics, and probiotics. Today, the use of antibiotics with natural ingredients is becoming a trend. One of the natural ingredients that contain high antioxidants and antibiotics is sp. This research was conducted from March to May 2022 at the Biotechnology Laboratory, Faculty of Fisheries and Marine, Universitas Riau, in two stages: 1) the sensitivity of extracts of sp. and 2) the application of sp. extract orally in tilapia ( ). The parameters measured were clear zone, minimum inhibitory concentration, LD test of leaf extract of sp. in tilapia ( ), hemoglobin levels, hematocrit, total leukocytes, total erythrocytes, leukocyte differentiation, and survival rate. Data on hematology parameters were tabulated and analyzed using a One-Way ANOVA followed by a Student Newman Keuls (SNK) test when deemed necessary. The results showed that the extract of sp. inhibited the growth of bacteria with a clear zone of 6.5-15.0 mm, which is classified as resistant. At doses of 2000, 2500, and 3000 ppm, it did not cause death in fish for 96 hours (LD ). Hematological parameters can be a sign of the health status of fish. Tilapia given sp. in different doses gave an effect between treatments, both after 30 days of rearing and post-test against bacteria (p<0.05). The results showed that the hematology of fish fed with sp. extract was in the normal or healthy range. Healthy tilapia had erythrocyte counts ranging from 1.34-2.11×10 cells/mm , hematocrit 26.17-33.19%, hemoglobin 6.26-11.2 g/dL and total leukocytes 1.01-1.50×10 cells/mm and total erythrocytes 5.88-9.13×10 cells/ mm . A dose of 3000 ppm provided the highest health improvement against bacterial infection.
Topics: Animals; Sargassum; Cichlids; Aeromonas hydrophila; Microbial Sensitivity Tests; Anti-Bacterial Agents; Fish Diseases
PubMed: 38817412
DOI: 10.12688/f1000research.128819.2 -
Comparative Biochemistry and... May 2024Cathelicidins are important antimicrobial peptides in various vertebrate species where they are crucial parts of the innate immune system. The current understanding of...
Cathelicidins are important antimicrobial peptides in various vertebrate species where they are crucial parts of the innate immune system. The current understanding of amphibian cathelicidins is limited, particularly with regard to their immunomodulatory effects. To address this knowledge gap, we produced the cDNA sequence of the cathelicidin gene from a skin transcriptome of the Chinese spiny frog Quasipaa spinosa. The amino acid sequence of the Quasipaa spinosa cathelicidin (QS-CATH) was predicted to consist of a signal peptide, a cathelin domain, and a mature peptide. Comparative analysis of the QS-CATH amino acid sequence with that of other amphibian cathelicidins revealed high variability in the functional mature peptide among amphibians, whereas the cathelin domain was conserved. The QS-CATH gene was expressed in several tissues, with the highest level of expression in the spleen. Upregulation of QS-CATH after Aeromonas hydrophila infection occurred in the kidney, gut, spleen, skin, and liver. Chemically synthesized QS-CATH exhibited pronounced antibacterial activity against Shigella flexneri, Staphylococcus warneri, Escherichia coli, Salmonella enterica, and Listeria monocytogenes. Furthermore, QS-CATH disrupted the cell membrane integrity of S. flexneri, as evidenced by a lactate dehydrogenase release assay, and it hydrolyzed the genomic DNA of S. flexneri. Additionally, QS-CATH elicited chemotaxis and modulated the expression of inflammatory cytokine genes in RAW264.7 mouse leukemic monocyte/macrophage cells. These findings confirm the antimicrobial effects of amphibian cathelicidin and its ability to influence immune cell function. This will expedite the potential utilization of amphibian antimicrobial peptides as therapeutic agents.
PubMed: 38810897
DOI: 10.1016/j.cbpc.2024.109943