Search Results (132)

Mori Cortex Radicis (MCR), which is abundant in resources and low in cost, is a Chinese herbal medicine with antitussive, anti-inflammatory, analgesic, and hypoglycemic effects; however, its application in the prevention and control of aquatic pathogens remains understudied. In this study, a MCR water extract (MCR-WE) was prepared, and its contents of polysaccharides, polyphenols, and proteins were found to be 0.63%, 1.17%, and 2.79%, respectively. LC-MS metabolomics revealed that L(+)-Arginine, 9,12,13-Todea, Citric acid, 1-Deoxynojirimycin, and 4-Guanidinobutanoic acid were the most abundant compounds. Subsequently, by feeding the MCR-WE to crucian carp (Carassius auratus) and challenging them with Aeromonas veronii, it was found that the MCR-WE enhanced the activities of immune factors (AKP, ACP, LZM) and the phagocytic activity of leukocytes (p < 0.05). Furthermore, the MCR-WE improved the survival rate of crucian carp (p < 0.05), reduced the bacterial load in the kidneys (p < 0.05), decreased the mRNA expression of inflammatory factors (IL-6, IL-1β, TNF-α), and lowered the expression levels of antioxidant-related factors (CAT, GSH-Px, SOD, MDA) and the mRNAs of oxidative stress pathway factors (Nrf2, HO-1, Keap1) (p < 0.05). Histopathological sections and immunofluorescence assays showed that the MCR-WE maintained the structural integrity of internal organs and reduced renal cell apoptosis and DNA damage. Therefore, MCR-WE is rich in immunologically active substances, can activate the immune response of crucian carp, reduce fish mortality, exert anti-inflammatory and antioxidant activities, and maintain the structural and functional integrity of internal organs. Thus, the MCR-WE holds promise as a therapeutic agent against A. veronii infection in fish. Full article

Background/Objectives: Environmental dissemination of antimicrobial resistance (AMR) is increasingly driven by wastewater-impacted aquatic systems, yet the key factors controlling multidrug-resistant (MDR) bacterial distribution remain unclear. This study evaluated environmental factors associated with MDR bacteria in the urban Danube River (Novi Sad, Serbia) using a machine learning framework. Methods: Surface-water and wastewater samples were collected during summer and autumn 2024. Bacterial isolates were obtained through membrane filtration onto chromogenic media and identified using MALDI-TOF MS. Physicochemical parameters (including COD, BOD₅, turbidity, pH, and temperature) were used as predictors in seven machine learning models (ANN, RF, SVM, XGB, MARS, TREE, NB). Model performance was assessed using AUC, accuracy, and error metrics. Results: Wastewater samples showed higher bacterial abundance and taxonomic richness than river surface-water samples, with frequent recovery of Klebsiella pneumoniae, Escherichia coli, Aeromonas veronii, and Pseudomonas spp. Tree-based models (RF, XGB) performed best. Organic pollution indicators, turbidity, pH, and water temperature were the most prominent factors. Conclusions: Wastewater-related pollution gradients, reflected by organic load parameters, turbidity, pH, and water temperature, were associated with the occurrence of selected bacterial taxa recovered on selective media. These associations were more pronounced in wastewater samples, while river surface-water samples showed lower abundance and taxonomic richness but still contained selected environmental and opportunistic taxa. Because antimicrobial susceptibility testing and molecular confirmation of resistance determinants were not performed, the findings should be interpreted as culture-based and exploratory. Machine learning approaches may support environmental screening and hypothesis generation for AMR-oriented surveillance, but future studies should include standardized phenotypic and molecular confirmation. Full article

Wastewater treatment plants represent a primary source of environmental dissemination of multidrug-resistant (MDR) bacteria, underscoring the urgent need for in-depth investigation of these organisms. While the resistome of MDR bacteria has been extensively studied, there remains a critical gap in understanding the role of plasmid-borne genes encoding adaptive metabolic functions. We isolated two MDR strains from municipal wastewater, Klebsiella sp. KOS9 and Pseudomonas veronii Yu15, both exhibiting resistance to antibiotics, including ampicillin, cefazolin, kanamycin, streptomycin, erythromycin, chloramphenicol, tetracycline, and ciprofloxacin. The plasmids of these strains harbored genes encoding aliphatic amidases, as well as antibiotic resistance genes (ARGs) and enzymes involved in glycogen and dTDP-L-rhamnose biosynthesis, which may contribute to virulence. In Klebsiella sp. KOS9 a single acetamidase operon, was found on the megaplasmid, along with copper and silver resistance genes. P. veronii Yu15 harbored an operon containing the acetamidase and formamidase genes on the chromosome, as well as a phylogenetically distant acetamidase operon on the conjugative megaplasmid. Both strains exhibit acetamidase activity and P. veronii Yu15 was able to utilize acetamide and formamide as sole nitrogen sources. The occurrence of ARGs and adaptive accessory genes on plasmids likely enhances the competitiveness and environmental flexibility of these MDR bacteria. Full article

Aquaponic systems are increasingly recognized as sustainable technologies for integrated fish and vegetable production. However, concerns remain regarding the potential internalization of human pathogens into vegetables grown in these systems. This study assessed the risk of pathogen internalization in lettuce leaves grown in aquaponic systems with Nile tilapia challenged with Escherichia coli or Vibrio cholerae. The system comprised nine fish tanks, eighteen hydroponic pipes, and eighty-one lettuce plants, with tanks assigned to three treatments. Samples of water, fish gut, fish blood, and lettuce leaves were collected. Microbiological analyses included selective culture, biochemical assays, and molecular identification. Although colonies consistent with E. coli and V. cholerae were recovered on selective media, molecular sequencing identified other bacterial species, including Aeromonas sp., Aeromonas caviae, Aeromonas veronii, Enterobacter hormaechei, and Citrobacter freundii. The findings indicate that conventional culture-based methods may produce false-positive results and highlight the importance of molecular confirmation. Notably, pathogenic bacteria associated with tilapia were detected and appeared capable of disseminating through the system and internalizing into lettuce tissues. This result highlights the need for biosecurity measures, contamination monitoring, and the combined use of conventional and molecular diagnostic tools to ensure accurate pathogen detection and compliance with international food safety standards. Full article

Background: Aeromonas veronii is an important bacterial pathogen in crucian carp and can cause serious disease outbreaks and substantial economic losses in aquaculture. Objectives: To evaluate how A. veronii infection and its inactivated vaccine modulate immune responses in Carassius auratus. Methods: 270 juveniles were allocated into three groups: a saline-injected control group (Ctrl), a vaccination group receiving an inactivated A. veronii vaccine (Vac), and an artificial infection group (AIG) subjected to stimulation. Liver, spleen, head kidney, gill, and intestine samples were collected from fish after anesthesia. The relative transcript levels of IgM, IgD, BAFF, MHCII, CD4, BCL6, MyD88, and NF-κB were quantified. For liver transcriptome analysis, the effective library concentration was determined. And the 16S rRNA gene resulting reads of fish gill symbiotic microbiota were processed for downstream bioinformatic analysis. Results: The results showed that the Vac achieved an RPS of 73.33%, and vaccination significantly upregulated multiple immune-related genes in different fish organs. With BAFF transcription across organs emerging as a robust sentinel readout. The Pearson correlation coefficient (r) of BAFF between other genes were all ≥0.8. GO and KEGG enrichment analyses indicated that AIG had more DEGs than Vac (5885 vs. 4008) and Ctrl (6910 vs. 6178), respectively. Some genes in AIG revealed significant over-representation of immune pathways, such as BCL6, MyD88, and NF-κB. The fish gill microbiota comprised a diverse set of low-abundance taxa, the phylum level was dominated by Proteobacteria and Fusobacteriota across all groups; whereas, the Vac group remained broadly closer to the Ctrl group in overall composition. Conclusions: These results indicated marked post-challenge immune–metabolic coupling in the liver, and suggested coordinated immunophysiological interplay between the liver and the spleen. Gill microecology of symbiotic bacteria was affected by vaccination or challenge reactions, which in turn affects the health of the gills or the organism itself. Full article

This pioneering study investigates bacteria isolated from marine pelagic fish, fishing vessels, and gear surfaces, focusing on the variability in biofilm formation across different substrates, media, and cultivation conditions. Bacteria from fish intestines, skin, and gills, including spoilage organisms and potential fish and human pathogens, can contaminate vessel surfaces, gear, and containers and may act as microbial reservoirs and transmission vectors. In this study, biofilm formation was evaluated at air–liquid interfaces and on submerged plastic, metal, and glass surfaces under various incubation temperatures and media. Vibrio spp. were isolated both from fishing nets and fish gills, particularly Vibrio alginolyticus, V. gigantis, and V. pelagius. Although V. harveyi was examined as a representative vibrio, it did not form a biofilm on polypropylene. Photobacterium damselae subsp. damselae, Pseudomonas fragi, P. gessardii, Psychrobacter spp., and Rothia endophytica showed a strong affinity for stainless steel. Overall adhesion regardless of media type was highest for P. gessardii, followed by P. damselae and Aeromonas veronii, which adhered strongly to steel, glass, and polypropylene; however, only P. gessardii also adhered well to polystyrene, an important finding because these are known fish and human pathogens. These results highlight species-dependent biofilm triggers and their substantial variability and provide guidance for standardized marine biofilm protocols. Full article

Toll-like receptors (TLRs) are essential pattern recognition receptors of the innate immune system and play critical roles in pathogen invasion in teleosts. In this study, we identified and characterized full-length open reading frames of three TLRs belonging to the TLR11 subfamily from Schizothorax prenanti, termed spTLR19 (2868 bp), spTLR20 (2835 bp), and spTLR21 (2946 bp), encoding 955, 944, and 981 amino acids, respectively. All three proteins exhibited the conserved domain architecture typical of TLRs, comprising a leucine-rich repeat (LRR) domain, a transmembrane region, and a Toll/IL-1 receptor (TIR) domain. Phylogenetic and homology analyses revealed that spTLR19 and spTLR20 clustered most closely with their homologues from Cyprinus carpio, while spTLR21 showed the highest similarity to Onychostoma macrolepis TLR21. Expression profiling showed that these TLRs were ubiquitously expressed across examined tissues, with relatively higher expression in immune-related tissues such as spleen and gills. Furthermore, challenge with Streptococcus agalactiae and Aeromonas veronii significantly up-regulated the expression of spTLR19, spTLR20, and spTLR21 in spleen, liver, and gills, suggesting their involvement in antibacterial immune responses. These findings enhance the functional understanding of the teleost TLR11 subfamily and provide a foundation for elucidating disease resistance and immune regulation in S. prenanti. Full article

Aquarium fish are increasingly being recognized as reservoirs of zoonotic pathogens, with Aeromonas species posing a notable risk because of their environmental resilience and opportunistic pathogenicity. This study presents the most comprehensive genome-based investigation to date of Aeromonas diversity in aquarium fish, aiming to characterize their taxonomic distribution, population structure, and genomic features. A total of 64 Aeromonas isolates were collected from various aquarium fish species. Wholegenome sequencing was conducted on all isolates to facilitate comparative genomic analyses. Key approaches included multilocus sequence typing (MLST), pairwise Digital DNA-DNA hybridization (dDDH), and the construction of a phylogenomic tree for species-level classification. Furthermore, a population structure analysis was performed to explore genomic diversity and evolutionary trends among the isolates. The results identified 14 distinct Aeromonas species, with A. veronii, A. caviae, and A. hydrophila being the most common. Importantly, several isolates exhibited taxonomic ambiguity, indicating the possible presence of new species or subspecies lineages. Furthermore, antimicrobial resistance gene profiles and virulence factor distributions varied significantly across clades, indicating genomic plasticity. This study highlights the underappreciated genomic complexity of Aeromonas populations in aquarium environments and raises concerns about the public health implications of pathogen reservoirs in ornamental fish. Full article

Co-infections are increasingly recognized as drivers of disease in ornamental fish, yet their genomic underpinnings and zoonotic implications remain underexplored compared to farmed species. Leveraging a One Health perspective, we investigated an acute mortality event in koi carp and characterized a co-infection by opportunistic aquatic bacteria that are also implicated in human disease. We isolated Aeromonas veronii and Shewanella sp. from a moribund koi using culture, biochemical assays, and MALDI-TOF MS, then generated draft genomes and performed orthology (OrthoVenn3), pathway annotation (KEGG BlastKOALA/Mapper), secondary-metabolite mining (antiSMASH), and virulence/resistome screening (VFDB/CARD), complemented by antimicrobial susceptibility testing. Clinically, affected fish showed dropsy/ascites, scale loss, abnormal buoyancy, and reduced activity. Phylogenomics positioned A. veronii Koi-2 within the A. veronii complex near species thresholds (ANI ~96.1%; dDDH ~70.2%), while Shewanella sp. Koi-1 formed a distinct lineage below accepted cut-offs relative to S. seohaensis (ANI ~95.9%; dDDH ~67.6%). The virulome comprised 194 loci in A. veronii Koi-2 and 152 in Shewanella sp. Koi-1 is enriched for adhesion, secretion, iron uptake, and immune-evasion functions. Genotype–phenotype agreement was high for multidrug resistance: Shewanella sp. encoded OXA-436 and rsmA, matching β-lactam resistance and reduced fluoroquinolone/phenicol susceptibility, whereas A. veronii carried tet(A), OXA-1157, cphA3, sul1, and aadA3 consistent with tetracycline, β-lactam, sulfonamide, and aminoglycoside resistance profiles. In conclusion, genome-resolved diagnostics confirmed a mixed Aeromonas–Shewanella co-infection with broad virulence potential and convergent resistance mechanisms, supporting the routine use of genomics to distinguish single- versus mixed-agent disease and to guide dual-coverage, mechanism-aware therapy in ornamental fish medicine while informing zoonotic risk mitigation. Full article

Intensive aquaculture and animal farming along riverbanks have emerged as significant drivers of downstream public health risks, facilitating the transmission of zoonotic pathogens and antimicrobial resistance (AMR) genes from farm effluents into natural water systems. In this study, we conducted a comprehensive 12-week water monitoring program at the Wei River in Shandong, China, using a combination of rapid detection techniques (RPA-LFD) and whole-genome sequencing to trace the origins of detected pathogens. RPA-LFD screening revealed the sequential appearance of Vibrio parahaemolyticus, Aeromonas veronii, norovirus GII, and Brucella spp. in surface water from March onward, coinciding with documented wastewater discharge events from upstream shrimp and fox farms. Subsequent isolation efforts confirmed the presence of V. parahaemolyticus and A. veronii in both river water and shrimp samples, while Brucella abortus was isolated from fox feces and water samples. Whole-genome sequencing of bacterial isolates revealed that V. parahaemolyticus strains from water and shrimp shared identical sequence types (ST150 and ST809) and resistance gene profiles, indicating a clonal relationship. Similarly, B. abortus isolates from water and fox feces differed by fewer than five SNPs, confirming farm-to-water transmission. Norovirus GII.3 and GII.6 sequences from water and fecal samples clustered phylogenetically with regional clinical strains, suggesting local circulation and environmental dissemination. Our findings highlight the critical role of river water monitoring as an early warning system for pathogen spread, emphasizing the need for integrated surveillance systems that monitor both water quality and the health of upstream farms and wildlife populations. The combined use of RPA-LFD and whole-genome sequencing provides a robust framework for real-time detection and source tracing of zoonotic pathogens, offering valuable insights for future environmental monitoring and public health interventions. Full article

The genus Aeromonas is widely distributed in aquatic environments, where it is a frequent fish pathogen. It has also been described in association with human infections, with most cases caused by A. caviae, A. veronii biovar sobria, and A. hydrophila. More recently, A. dhakensis has emerged as an increasingly important human pathogen. Transmission occurs primarily through ingestion or contacts with aquatic sources, or by consuming contaminated food, particularly from aquatic origins. Growing resistance in Aeromonas has been reported for penicillins (including their combinations with classical β-lactamase inhibitors), cephalosporins, and carbapenems. Among the β-lactam antibiotics, only fourth-generation cephalosporins remain almost uniformly active. Furthermore, the co-occurrence of resistance genes for third-generation cephalosporins and carbapenems within the same isolates is increasing. Recently, the presence of mobile genes conferring colistin resistance has also been documented, with resistance rates sometimes exceeding 30%. This evolution of colistin resistance is likely linked to its use in aquaculture, and together with the rise in β-lactam resistance, may be transforming Aeromonas into a significant reservoir of resistance genes that could potentially be transferred to species more commonly associated with human infections, such as the Enterobacterales. Full article

Aeromonas veronii is a major fish pathogen that causes substantial economic losses in fish aquaculture. The objective of this study was to isolate and characterize bacteriophages with prospects for the biocontrol of A. veronii in aquaculture as an alternative to conventional antibiotics. Phages were isolated from different wastewater sources and screened for performance using a tiered approach. The top two phages, DJ6712 and FW6709, were characterized using host range assays, transmission electron microscopy (TEM), temperature and pH stability assays, optimal multiplicity of infection (MOI) assays, and one-step growth curves. DJ6712 and FW6709 were only specific to A. veronii and infected 84% and 72% of the A. veronii isolates tested, respectively. TEM showed that DJ6712 and FW6709 belong to the family Siphoviridae. Both phages showed high host bacterial growth inhibition at MOI of 1. DJ6712 demonstrated higher temperature (30–50 °C) and pH tolerance (5–8) compared to FW6709, thus making it a more robust candidate for prospective biocontrol against A. veronii as an alternative to antibiotics in aquaculture. The study outcomes could offer an excellent addition to the existing global phage arsenal against A. veronii and expand the limited knowledge on A. veronii phages as an early preparation against this emerging threat. Full article

During four outbreaks in 2023 and 2024, samples from pond-reared Nile tilapia were taken from different farms located in Kafr Elsheikh governorate, Egypt. Samples were submitted for laboratory examinations. Diseased fish exhibited bacterial septicemia and some cases died without showing any clinical signs. A total of 30 bacterial isolates were isolated and identified. Of these isolates, 57% were identified as Gram-positive bacteria, whereas the remaining 43% were identified as Gram-negative bacteria. PCR targeting the 16S rRNA gene and genome sequencing confirmed five bacterial isolates as Aeromonas veronii (30%), Vibrio alginolyticus (13.3%), Enterococcus faecalis (23.3%), Aerococcus viridans (16.7%), and Staphylococcus epidermidis (16.7%). The NCBI GenBank accession numbers of these strains were (PV018985) for A. veronii, (PV016854) for V. alginolyticus, (PV013413) for E. faecalis, (PV032005) for A. viridans, and (PV012491) for Staph. epidermidis. The antibiogram revealed that the bacterial strains showed resistance to most of the antibiotics tested. A. viridans exhibited resistance to nearly all the antibiotics except for intermediate sensitivity to ciprofloxacin and amoxycillin/clavulanic acid. However, A. veronii showed high sensitivity to amoxycillin/clavulanic acid, oxytetracycline, kanamycin, and trimethoprim/sulfamethoxazole and intermediate susceptibility to ciprofloxacin. Similarly, E. faecalis showed high susceptibility to amoxycillin/clavulanic acid, ciprofloxacin, and trimethoprim/sulfamethoxazole in addition to intermediate sensitivity to ampicillin and kanamycin. Furthermore, Staph. epidermidis strain was highly susceptible to ampicillin, amoxycillin/clavulanic acid, oxytetracycline, novobiocin, and trimethoprim/sulfamethoxazole and was partially sensitive to kanamycin and ciprofloxacin. To conclude, summer mortalities recorded in farmed tilapia were closely related to a multifactorial bacterial origin with different sensitivity to antibiotic discs. Full article

Aeromonas veronii is a major pathogen threatening freshwater aquaculture, yet the molecular mechanisms of Pelteobagrus vachellii’s immune response to this infection remain unclear. This study integrated histopathology, mRNA-seq and small RNA-seq to investigate P. vachellii’s response to A. veronii at 48 h post-challenge. Histopathologically, infection induced gill epithelial detachment, hepatocyte swelling with cytoplasmic vacuolation, and melanomacrophage centers (MMCs) in the mid-kidney (histological assessment of the head kidney was not feasible due to sampling limitations associated with its small size). Transcriptomic analysis identified 1210 differentially expressed genes (DEGs) in the head kidney (819 downregulated, 391 upregulated), significantly enriched in 11 immune pathways (e.g., NF-κB, Th17 cell differentiation, Complement and coagulation cascades), with key immune genes (e.g., IL-1β, TCRα, CCL4) upregulated. Gene Set Enrichment Analysis (GSEA) revealed activation of the proteasome, ribosome and oxidative phosphorylation pathways, and suppression of the autophagy-animal, FoxO and AMPK pathways. Small RNA-seq identified 544 known and 958 novel miRNAs in the head kidney, with 42 downregulated and 36 upregulated differentially expressed miRNAs (DE miRNAs). The miRNA-mRNA network showed that DE miRNAs (e.g., miR-101-y/z, miR-132-z, miR-3167-y) negatively regulated immune-related target genes (IL-1R1, IRF4, IκBα) in core immune pathways. Collectively, this study clarifies the pathological and miRNA-mRNA regulatory modules of P. vachellii head kidney against A. veronii infection, providing valuable information that enables the further analyses of the defense mechanisms of P. vachellii against A. veronii infection. Full article

Egg yolk IgY antibody has significant application potential in aquaculture as a form of passive immunotherapy against various bacterial infections owing to its capacity for large-scale and cost-effective production. In this research, laying hens were immunized with live or inactivated Aeromonas hydrophila to produce IgY antibodies. Following this, experiments were carried out to assess the passive immune protection rates of the two types of IgY antibodies when used to immunize goldfish (Carassius auratus), which were then infected with A. hydrophila or Aeromonas veronii. ELISA experiments were conducted to demonstrate the interaction between the IgY antibodies and the bacteria. The kidneys of C. auratus were coated on a Luria–Bertani (LB) medium to evaluate bacterial content. The leukocyte phagocytosis was detected by a cell phagocytosis assay. The serum of C. auratus was used to assess the expression of antioxidant factors, and a qRT-PCR was conducted to evaluate the mRNA expression of inflammatory factors in visceral tissue. Furthermore, histopathology and immunofluorescence analysis were performed to evaluate the structural integrity, apoptosis, and DNA damage of visceral tissues. The results indicated that the live or inactivated A. hydrophila IgY antibodies exhibited passive immune protection rates against A. hydrophila and A. veronii and could recognize these two bacteria in vitro. Additionally, these two IgY improved the phagocytic ability of leukocytes, diminished renal bacterial concentration, and decreased the levels of antioxidant factors and mRNA expression of inflammatory factors. Meanwhile, the two IgY antibodies did not cause any pathology of the kidney, spleen, and intestine, and decreased the levels of DNA damage factor (γH2A.X) and cell apoptosis factor (p53) in renal tissue. Therefore, live and inactivated A. hydrophila IgY antibodies can resist bacterial infections, with live bacteria IgY providing greater protection than inactivated bacteria IgY. Further, A. hydrophila is an aquatic pathogen that causes minimal damage to laying hens, and the immunity of live A. hydrophila conforms to animal welfare. Altogether, live A. hydrophila IgY antibody can serve as a polyvalent passive immune vaccine candidate in aquaculture. Full article