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Search Results (1,170)

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16 pages, 1811 KB  
Article
Nanopore-Based Metagenomic Approaches for Detection of Bacterial Pathogens in Recirculating Aquaculture Systems
by Diego Valenzuela-Miranda, María Morales-Rivera, Jorge Mancilla-Schutz, Alberto Sandoval, Valentina Valenzuela-Muñoz and Cristian Gallardo-Escárate
Fishes 2025, 10(10), 496; https://doi.org/10.3390/fishes10100496 - 2 Oct 2025
Abstract
The microbial community in a recirculating aquaculture system (RAS) is pivotal in fish health, contributing significantly to the productive performance during the growing-out phase. Classical and molecular methods using PCR for species-specific amplifications have traditionally been used for bacterial community surveillance. Unfortunately, these [...] Read more.
The microbial community in a recirculating aquaculture system (RAS) is pivotal in fish health, contributing significantly to the productive performance during the growing-out phase. Classical and molecular methods using PCR for species-specific amplifications have traditionally been used for bacterial community surveillance. Unfortunately, these approaches mask the real bacterial diversity and abundance, population dynamics, and prevalence of pathogenic bacteria. In this study, we explored the use of Oxford Nanopore Technology to characterize the microbiota and functional metagenomics in a commercial freshwater RAS. Intestine samples from Atlantic salmon (Salmo salar (85 ± 5.7 g)) and water samples from the inlet/outlet water, settling tank, and biofilters were collected. The full-length 16S rRNA gene was sequenced to reconstruct the microbial community, and bioinformatic tools were applied to estimate the functional potential in the RAS and fish microbiota. The analysis showed that bacteria involved in denitrification processes were found in water samples, as well as metabolic pathways related to hydrogen sulfide metabolism. Observations suggested that fish classified as sick exhibited decreased microbial diversity compared with fish without clinical symptomatology (p < 0.05). Proteobacteria were predominant in ill fish, and pathogens of the genera Aeromonas, Aliivibrio, and Vibrio were detected in all intestinal samples. Notably, Aliivibrio wodanis was detected in fish showing abnormal clinical conditions. Healthy salmon showed higher contributions of pathways related to amino acid metabolism and short-chain fatty acid fermentation (p < 0.05), which may indicate more favorable fish conditions. These findings suggest the utility of nanopore sequencing methods in assessing the microbial community in RASs for salmon aquaculture. Full article
(This article belongs to the Special Issue Infection and Detection of Bacterial Pathogens in Aquaculture)
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14 pages, 3946 KB  
Article
Histopathological and Molecular Insights into Grass Carp Kidney Responses to Co-Infection with Aeromonas hydrophila and Aeromonas veronii
by Yifei Zhou, Ruijun Zhu, Lingli Xie, Wenyao Lv, Xinyue Wang, Mengzhou Wu, Xiaoyan Xu and Junqiang Qiu
Fishes 2025, 10(10), 484; https://doi.org/10.3390/fishes10100484 - 29 Sep 2025
Abstract
Grass carp (Ctenopharyngodon idella), a key species in freshwater aquaculture, is particularly vulnerable to opportunistic pathogens, including Aeromonas hydrophila and Aeromonas veronii. While the pathogenic mechanisms of individual infections have been extensively characterized, the host immune responses during co-infection remain [...] Read more.
Grass carp (Ctenopharyngodon idella), a key species in freshwater aquaculture, is particularly vulnerable to opportunistic pathogens, including Aeromonas hydrophila and Aeromonas veronii. While the pathogenic mechanisms of individual infections have been extensively characterized, the host immune responses during co-infection remain poorly understood. This research explored the renal pathological alterations and transcriptomic shifts in grass carp subjected to simultaneous infection by A. hydrophila and A. veronii. Mortality occurred as early as 24 h post-challenge, ultimately reaching a cumulative death rate of 65%. Quantitative analysis of renal bacterial burden revealed a marked increase in colonization at 3 days post-infection (dpi). The histopathological assessment showed progressive kidney damage, including tubular collapse, epithelial necrosis, interstitial edema, and widespread epithelial desquamation, with the most severe lesions observed at 5 dpi and partial signs of recovery by 7 dpi. A total of 1106 and 472 genes were found to be differentially expressed at 1 and 5 dpi, respectively, based on transcriptome profiling. The functional enrichment analysis indicated that the differentially expressed genes (DEGs) were mainly involved in the complement and coagulation cascade pathways. Notably, the immune-related genes exhibited a biphasic trend, with predominant downregulation at 1 dpi followed by marked upregulation by 5 dpi, indicating dynamic changes in immune modulation during co-infection. These results provide new insights into host responses during dual bacterial infections in fish and may inform disease prevention strategies in aquaculture. Full article
(This article belongs to the Section Welfare, Health and Disease)
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22 pages, 3026 KB  
Article
One Health Monitoring of Resistant Pseudomonas aeruginosa in Aquatic, Poultry, and Human Sources: Virulence Traits and blaSHV Gene Tracking
by Ali Wahdan, Mahmoud Ezzat, Amal Emam, Walaa A. Husseiny, Mohamed Abou El-Atta, Ehab M. Abd-Allah, Ahmed M. A. Meligy, Sherief M. Abdel-Raheem, Faisal Almathen, Salah Al-Shami, Saad I. Al-Sultan, Ahmed Alfifi, Wael El-Deeb and Marwa E. Abo Hashem
Pathogens 2025, 14(10), 983; https://doi.org/10.3390/pathogens14100983 - 28 Sep 2025
Abstract
Pseudomonas aeruginosa is an opportunistic pathogen of public health concern. This study aimed to investigate the prevalence of P. aeruginosa, some virulence factors, and antimicrobial resistance patterns and highlight the potential pathways of horizontal blaSHV-resistant gene transfer from diverse sources. [...] Read more.
Pseudomonas aeruginosa is an opportunistic pathogen of public health concern. This study aimed to investigate the prevalence of P. aeruginosa, some virulence factors, and antimicrobial resistance patterns and highlight the potential pathways of horizontal blaSHV-resistant gene transfer from diverse sources. A total of 220 samples were collected from fish (n = 90), water (n = 30), poultry (n = 50), and humans (n = 50). All samples were isolated, confirmed by the Vitek 2 system, and tested against antimicrobial agents. Some virulence and resistance genes were examined by PCR and sequenced for the blaSHV-resistant gene from four selected isolates from each source. SPSS v26, with chi-squared tests and Pearson correlations (p < 0.05), was implemented for statistical investigation. P. aeruginosa was isolated at 33.3%, 20%, 14%, and 24% from fish, water, poultry, and humans, respectively. Using the diffusion disk method, extensively drug-resistant (XDR) and multidrug-resistant (MDR) strains were detected. All strains harbored the oprL and toxA genes, while the lasB gene was present in 40% of fish samples but not present in human samples. All strains lacked the exoS gene. The tetA, sul1, blaSHV, and blaTEM resistance genes were detected at different percentages. The blaSHV genes from fish and water isolates were closely related to each other and showed similarity to those of the human isolates. The poultry isolates formed a separate phylogenetic lineage. The emergence of XDR and MDR P. aeruginosa highlights a possible public health threat. Based on the gene similarity between fish and water isolates, our results suggest that these isolates have a common origin. The similarity between the human isolates and environmental isolates (fish and water) raises concerns about possible transmission to humans. Full article
(This article belongs to the Section Bacterial Pathogens)
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41 pages, 7341 KB  
Article
Dynamics of Pathomorphological and Pathophysiological Alterations in Rainbow Trout (Oncorhynchus mykiss) During Acute Aeromonas salmonicida Infection
by Dmitry Nikiforov-Nikishin, Nikita Kochetkov, Kirill Gavrilin, Viktoria Gaffarova, Kirill Medvedev, Svetlana Smorodinskaya, Anastasia Klimuk, Yuri Kuchikhin, Ivan Svinarev, Natalya Gladysh, Anna Kudryavtseva, Egor Shitikov and Alexei Nikiforov-Nikishin
Biology 2025, 14(10), 1330; https://doi.org/10.3390/biology14101330 - 26 Sep 2025
Abstract
Furunculosis caused by Aeromonas salmonicida is one of the most common diseases in aquaculture, leading to significant economic losses. This study comprehensively investigated the dynamics of pathophysiological and histopathological disorders in rainbow trout (Oncorhynchus mykiss) infected with the moderately virulent strain [...] Read more.
Furunculosis caused by Aeromonas salmonicida is one of the most common diseases in aquaculture, leading to significant economic losses. This study comprehensively investigated the dynamics of pathophysiological and histopathological disorders in rainbow trout (Oncorhynchus mykiss) infected with the moderately virulent strain A. salmonicida SL0n. Whole-genome analysis showed that strain SL0n belongs to the A. salmonicida species complex, possessing a single circular chromosome. The genome encodes a wide range of virulence factors, including adhesion systems (type IV pili, fimbriae), toxins (aerolysin, hemolysins), and a type II secretion system (T2SS), but notably lacks plasmids and a type III secretion system (T3SS). This genomic profile likely dictates a pathogenic mechanism reliant on secreted exotoxins (via T2SS), explaining the observed systemic cytotoxic damage. In an acute experiment, the 4-day LD50 was determined to be 1.63 × 106 CFU/fish. In a prolonged experiment, fish were injected with a sublethal dose (1.22 × 106 CFU/fish—75% of LD50). The disease progressed through three consecutive stages. The early stage (1–2 DPI) was characterized by maximal bacterial load and activation of nonspecific immunity. The acute stage (4 DPI) manifested as severe septicemia and anemia, associated with systemic organ damage, which correlated with peak AST and ALT enzyme activity. The recovery stage (6 DPI) was marked by partial regression of inflammation, key biochemical and histological parameters indicated persistent liver and kidney dysfunction, signifying an incomplete recovery. These results demonstrate the pathogenesis of acute furunculosis and reveal that the genomic profile of the SL0n strain causes a sequential, systemic infection characterized by severe organ dysfunction. Full article
(This article belongs to the Special Issue Aquatic Animal Pathogens and Immunity)
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40 pages, 3769 KB  
Article
Deciphering European Sea Bass (Dicentrarchus labrax) Resistance to Nervous Necrosis Virus by Transcriptome Analysis from Early Infection Towards Establishment of Virus Carrier State
by Dimitra K. Toubanaki, Odysseas-Panagiotis Tzortzatos, Antonia Efstathiou, Vasileios Bakopoulos and Evdokia Karagouni
Int. J. Mol. Sci. 2025, 26(18), 9220; https://doi.org/10.3390/ijms26189220 - 21 Sep 2025
Viewed by 255
Abstract
Viral nervous necrosis, caused by the nervous necrosis virus (NNV), is an important threat to aquaculture, causing great economic losses and a high environmental burden. European sea bass (Dicentrarchus labrax) is highly affected by NNV, and selective breeding programs for disease [...] Read more.
Viral nervous necrosis, caused by the nervous necrosis virus (NNV), is an important threat to aquaculture, causing great economic losses and a high environmental burden. European sea bass (Dicentrarchus labrax) is highly affected by NNV, and selective breeding programs for disease resistance have been established in order to achieve a sustainable aquaculture and minimize the need for vaccines, drugs and antibiotics. Resistant and susceptible European sea bass were experimentally challenged with NNV and their head kidney transcriptomes were analyzed at three time points, i.e., 3 hpi, 2 dpi and 14 dpi. Numerous differentially expressed genes (DEGs) were identified in the head kidneys of resistant and susceptible infected vs. non-infected sea bass. Gene ontology enrichment, pathway, and protein–protein interaction analyses revealed that the NNV-resistant fish control their response to viral infection more efficiently, utilizing different mechanisms compared to the susceptible fish. Resistant fish displayed higher levels of interferon-related elements, cytokines, antigen presentation, T-cell activity, apoptosis, and programmed cell death combined with a controlled inflammatory response and more active proteasome and lysosome functions. The susceptible fish appeared to have high immune responses at the early infection stages, accompanied by high expressions of inflammatory, complement and coagulation pathways. Insulin metabolism was better regulated in the resistant fish and the control of lipid metabolism was less effective in the susceptible family. The cytoskeleton- and cell adhesion-related pathways were mostly down-regulated in the susceptible fish, and the intracellular transport and motor proteins were utilized more efficiently by the resistant fish. The present study represents a thorough transcriptomic analysis of NNV infection effects on a resistant and a susceptible European sea bass head kidney. The obtained results provide valuable information on the mechanisms that offers pathogen resistance to a host, with many aspects that can be exploited to develop more efficient approaches to fighting viral diseases in aquaculture. Full article
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23 pages, 2053 KB  
Article
Integrated Omics-Based Discovery of Bioactive Halogenated Metabolites from the Deep-Sea Streptomyces sp. B188M101
by Emmanuel Tope Oluwabusola, Stephen A. Jackson, Cristina Brunati, Stefanie Gackstatter, Hannah Vedder, Marianna Iorio, Gargee Chawande, Lekha Menon Margassery, Giang-Son Nguyen, David J. Clarke, Rainer Ebel, Marcel Jaspars and Alan D. W. Dobson
Mar. Drugs 2025, 23(9), 362; https://doi.org/10.3390/md23090362 - 19 Sep 2025
Viewed by 509
Abstract
Using the one-strain-many-compounds (OSMAC) culturing approach, metabolomic studies, and bioassay-guided purification, we have isolated and characterised three new chlorinated natural products, agelolines B-D (13), together with two known compounds, ageloline A (4) and gausemycin A (5 [...] Read more.
Using the one-strain-many-compounds (OSMAC) culturing approach, metabolomic studies, and bioassay-guided purification, we have isolated and characterised three new chlorinated natural products, agelolines B-D (13), together with two known compounds, ageloline A (4) and gausemycin A (5), which have been identified by high-resolution mass spectrometry and 1D and 2D NMR analyses. The preliminary evaluation of three small-scale extracts (M400, R358 and SGG) against the fish pathogen, Aeromonas salmonicida subsp. achromogenes KELDUR265-87, showed that the R358 extract displayed significant activity. Furthermore, the natural products (15) were evaluated against the fish pathogen Aeromonas salmonicida and human pathogens (Stenotrophomonas maltophilia L2125, Staphylococcus aureus ATCC6538P, and S. pneumoniae L44) using a serial dilution assay. Compound 3 displayed activity against Staphylococcus aureus ATCC6538P, S. maltophilia L2125, and S. pneumoniae L44 with MIC values of 6, 32, and 64 µg/mL, respectively. Interestingly, only gausemycin A (5) exhibited considerable inhibition against A. salmonicida with an MIC value of 32 µg/mL, and the activity increased by two-fold when supplemented with 0.45 mM calcium salt, while 2 and 4 showed moderate inhibition against S. maltophilia L2125. The biosynthetic pathways of compounds 14 were proposed. This is the first report of specific inhibition of A. salmonicida by 5. Full article
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14 pages, 376 KB  
Review
Microbial Quality of Leafy Greens Grown Under Soilless Production Systems
by Robert Korir Cheruiyot and Abraham Fikru Mechesso
Pathogens 2025, 14(9), 943; https://doi.org/10.3390/pathogens14090943 - 18 Sep 2025
Viewed by 223
Abstract
This review examines the microbiological diversity and food safety implications of soilless production systems, particularly aquaponics and hydroponics, which are gaining popularity as efficient methods for producing fresh produce in controlled environments. Despite their advantages, a limited understanding of the microbiological quality and [...] Read more.
This review examines the microbiological diversity and food safety implications of soilless production systems, particularly aquaponics and hydroponics, which are gaining popularity as efficient methods for producing fresh produce in controlled environments. Despite their advantages, a limited understanding of the microbiological quality and potential food safety risks associated with leafy greens grown in these systems remains. By analyzing published studies, we summarize evidence of microbial contamination in aquaponic and hydroponic environments and their crops, noting that various factors may facilitate pathogen survival and spread to edible plant parts. The operational practices and environmental conditions can promote pathogen introduction through multiple routes, including contaminated fingerlings, fish feed, recirculating contaminated water, pest intrusion, improper handling, and poor worker hygiene. The studies reviewed detected pathogens that pose public health risks, including Salmonella spp., Listeria monocytogenes, Pseudomonas aeruginosa, and Shiga toxin-producing Escherichia coli O157:H7, as well as various molds. These potentially contaminated fresh produces are often consumed raw, presenting significant food safety and public health risks that demand further investigation and mitigation strategies to ensure consumer protection while maintaining the benefits of soilless agriculture. Full article
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18 pages, 9498 KB  
Article
Neuropeptide Y Boosts Intestinal Mucosal Immunity of Tilapia Infected with Streptococcus agalactiae by Reducing Inflammation and Oxidative Stress
by Yang Yu, Ziyan Liu, Mengyuan Zhou, Zexia Chen, Ran Cai, Chaowei Song, Meiqing Li, Tiansheng Zhu, Caiyun Sun and Wensheng Li
Animals 2025, 15(18), 2730; https://doi.org/10.3390/ani15182730 - 18 Sep 2025
Viewed by 222
Abstract
Neuropeptides and their corresponding receptors have been observed to play a significant role in modulating immune cell functions and epithelial barrier functions. In teleost, Neuropeptide Y (NPY) is identified to be involved in the immune regulation of intestinal tissues during bacterial infections. However, [...] Read more.
Neuropeptides and their corresponding receptors have been observed to play a significant role in modulating immune cell functions and epithelial barrier functions. In teleost, Neuropeptide Y (NPY) is identified to be involved in the immune regulation of intestinal tissues during bacterial infections. However, the mechanism of NPY on infected gastrointestinal tissue remains unclear, as does whether other members of the NPY family, like ligand PYYb and some receptors Y7 and Y8b, also contribute to this immunoregulatory process. Herein, the responses of PYYb and receptors Y7 and Y8b were explored after pathogen challenging, as well as the effect of NPY on the gastrointestinal tissues of Nile tilapia (Oreochromis niloticus). Using a NPY and S. agalactiae co-injection model, we measured the expression of pyyb, y7, and y8b and the inflammatory and oxidative stress responses in gastrointestinal tissues. S. agalactiae infection significantly upregulated pyyb expression, while co-treatment with NPY further amplified its expression. Infection caused an imbalance between y7 and y8b and was alleviated by NPY. Moreover, NPY contributed to immune protection. NPY co-treatment reduced inflammatory cytokine expression in the gastrointestinal tissues and alleviated tissue damage inflicted by S. agalactiae. Additionally, NPY mitigated immune and oxidative stress by enhancing lysozyme, AKP and ACP activities, normalizing glucose and albumin levels, and reducing lipid peroxidation (MDA). These findings demonstrated that PYYb and receptors Y7 and Y8b were involved in the immune response caused by S. agalactiae. NPY not only enhanced their reactions but also reduced inflammation and oxidative stress induced by the pathogen, indicating its potential as a therapeutic intervention for S. agalactiae infections in fish. Full article
(This article belongs to the Special Issue Fish Diseases and Healthy Farming)
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9 pages, 2434 KB  
Case Report
Traumatic Oral Lesions in Loggerhead Sea Turtles (Caretta caretta) Linked to Polychaete (Laetmonice cf. hystrix) Ingestion: A Case Report from the Northern Adriatic Sea
by Stefano Pesaro, Lucia Biagini, Danilo De Bellis, Luca Dorigo, Alice Baggio, Isabella Perlin and Giacomo Rossi
Animals 2025, 15(18), 2727; https://doi.org/10.3390/ani15182727 - 18 Sep 2025
Viewed by 264
Abstract
Oral cavity lesions in sea turtles, particularly Caretta caretta, are relatively rare, and are typically linked to infectious agents as well as anthropogenic factors, including ingestion of marine debris or fishing gear. This report describes a juvenile Caretta caretta found in the [...] Read more.
Oral cavity lesions in sea turtles, particularly Caretta caretta, are relatively rare, and are typically linked to infectious agents as well as anthropogenic factors, including ingestion of marine debris or fishing gear. This report describes a juvenile Caretta caretta found in the northern Adriatic Sea with severe oral lesions affecting the choanae, alimentary tract, and larynx. A comprehensive clinical and histopathological evaluation was conducted, which revealed traumatic injuries caused by the ingestion of a polychaete, Laetmonice cf. hystrix. Mucosal biopsies in the areas of spine penetration revealed the presence of strong sub-epithelial inflammation, characterised by micro-abscesses. In addition, around some fragments of spines, the formation of microgranulomatous lesions with a tendency to encapsulation was observed. Treatment protocol involved the removal of embedded spines and administration of a broad-spectrum antibiotic to prevent secondary infections. Recently, the detection in polychaetes of the aetiological agents of two newly emerging diseases of shrimps, suggests that these worms can be a host or/and passive carrier of these pathogens. This case study underscores the necessity to consider both biological and anthropogenic factors in diagnosing and managing oral lesions in marine turtles. Furthermore, it draws attention to the ecological risks posed by interactions between sea turtles and benthic organisms. Full article
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15 pages, 1808 KB  
Article
Strain-Dependent Thermoadaptation in the Fish Pathogen Aeromonas salmonicida subsp. salmonicida
by Kim C. Fournier, Pierre-Étienne Marcoux, Antony T. Vincent and Steve J. Charette
Microorganisms 2025, 13(9), 2171; https://doi.org/10.3390/microorganisms13092171 - 17 Sep 2025
Viewed by 282
Abstract
Strains of Aeromonas salmonicida subsp. salmonicida, a major pathogen of salmonids, typically do not grow at temperatures above 30 °C. The effects of thermal stress on this bacterium have been extensively studied. Recently, we demonstrated that repeated exposure to cyclical thermal stress, [...] Read more.
Strains of Aeromonas salmonicida subsp. salmonicida, a major pathogen of salmonids, typically do not grow at temperatures above 30 °C. The effects of thermal stress on this bacterium have been extensively studied. Recently, we demonstrated that repeated exposure to cyclical thermal stress, reaching up to 37 °C, can induce permanent thermoadaptation in certain strains of this bacterium. Many aspects of this adaptation process remain poorly understood. We generated 88 thermoadapted strains of A. salmonicida subsp. salmonicida capable of sustained growth at 33 °C or higher demonstrating that prolonged heat exposure can shift a substantial proportion of psychrophilic strains toward a more mesophilic-like behavior. Although growth at 35 °C was still weaker than in naturally mesophilic A. salmonicida strains, some thermoadapted strains were able to grow up to 37 °C. North American strains harboring the genomic island AsaGEI1a, a known biomarker, exhibited a significantly reduced capacity for thermoadaptation, suggesting a possible genetic constraint, although genomic analyses indicate that AsaGEI1a itself is not directly responsible for this limitation. Genotyping and phenotypic analyses revealed that thermoadaptation is frequently associated with the loss of Type III secretion system and the A-layer, two key virulence factors. Only 7% of the thermoadapted strains retained both features. Overall, our findings suggest that thermoadaptation may represent a potential mechanism influencing the persistence of some psychrophilic A. salmonicida subsp. salmonicida strains in warming aquatic environments under climate change. Full article
(This article belongs to the Section Veterinary Microbiology)
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14 pages, 1044 KB  
Review
Aquatic Chlamydiae: A Review of Their Roles in Fish Health
by Basma Mahmoud-Elkamouny, Carole Kebbi-Beghdadi and Gilbert Greub
Microorganisms 2025, 13(9), 2166; https://doi.org/10.3390/microorganisms13092166 - 17 Sep 2025
Viewed by 307
Abstract
Aquaculture plays a vital role in meeting the global demand for high-quality protein. However, the fish industry is challenged by infectious diseases, including gill conditions such as epitheliocystis. Epitheliocystis is characterized by cyst-like epithelial lesions, which occur in the gills of fish, and [...] Read more.
Aquaculture plays a vital role in meeting the global demand for high-quality protein. However, the fish industry is challenged by infectious diseases, including gill conditions such as epitheliocystis. Epitheliocystis is characterized by cyst-like epithelial lesions, which occur in the gills of fish, and is associated with intracellular bacteria including Chlamydia-related bacteria. Although epitheliocystis was initially regarded as of low significance, attention is increasing due to its impact on commercially important fish species in intense farming conditions. This review evaluates the roles of aquatic chlamydiae as pathogens contributing to fish morbidity and mortality, and as members of fish microbiota. Additionally, Chlamydia-related bacteria are thought to be involved in complex gill disease (CGD), characterized by lamellar fusion, epithelial hyperplasia, and inflammation. Recent discoveries have expanded the diversity of Chlamydiota isolated from fish, with novel species such as Candidatus (Ca.) Panilichlamydia rohitae, Ca. Piscichlamydia trichopodus, and Chlamydia vaughanii identified in different fish hosts. Most causative agents of epitheliocystis have not yet been cultured in vitro, although C. vaughanii, the first Chlamydiaceae member isolated from fish, was successfully cultured. As C. vaughanii was recently shown to be able to propagate in mammalian cells, it raises concerns about its zoonotic potential, although a pathogenic role has yet to be described. Full article
(This article belongs to the Section Veterinary Microbiology)
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13 pages, 2318 KB  
Article
Simultaneous Detection of Four Foodborne Pathogens in Raw Freshwater Fish Using High-Resolution Melting Analysis
by Shan Shan, Xiaoyu Tong, Wenyu Du, Yin Chen, Long Cheng, Fang Yan, Yujie Zhai, Kui Zhao, Haiyan Ni, Xiaomei Sha, Xiaoqing Liu, Chengwei Liu, Shuanglong Wang and Daofeng Liu
Foods 2025, 14(18), 3202; https://doi.org/10.3390/foods14183202 - 15 Sep 2025
Viewed by 359
Abstract
Many countries around the world feature raw fish in their cuisine, which is valued for its unique flavor. However, raw fish may be easily contaminated with foodborne pathogens including Listeria monocytogenes, Salmonella, Vibrio parahaemolyticus, and Staphylococcus aureus. Herein, a [...] Read more.
Many countries around the world feature raw fish in their cuisine, which is valued for its unique flavor. However, raw fish may be easily contaminated with foodborne pathogens including Listeria monocytogenes, Salmonella, Vibrio parahaemolyticus, and Staphylococcus aureus. Herein, a method was established that integrated a multiplex polymerase chain reaction (PCR) and high-resolution melting (HRM) curve assay for the simultaneous detection of these four foodborne pathogens. The target genes of the bacteria were amplified by PCR and subsequently analyzed using HRM. Differentiation was achieved based on the melting temperature (Tm) values of their respective amplicons. The detection limit of the PCR-HRM assay was 0.02–0.1 ng/µL. In addition, the Tm remained nearly constant across various concentrations of genomic DNA derived from the target bacteria. The assay demonstrated perfect specificity (8/8) and a sensitivity of 5/5 for L. monocytogenes, 2/2 for Salmonella, 3/3 for V. parahaemolyticus, and 3/3 for S. aureus. No significant interference occurred when genomic DNA from the four target bacteria was co-extracted with DNA from eight non-target strains. Furthermore, the assay offers advantages including operational simplicity, high efficiency, accurate results, reduced detection time, and lower costs, rendering it well-suited for food safety applications in the aquatic products processing industry. Full article
(This article belongs to the Section Food Microbiology)
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24 pages, 428 KB  
Review
Phytochemicals: Essential Oils and Other Extracts for Disease Prevention and Growth Enhancement in Aquaculture: Challenges and Opportunities
by Markos N. Kolygas, Konstantina Bitchava, Cosmas Nathanailides and Foteini Athanassopoulou
Animals 2025, 15(18), 2653; https://doi.org/10.3390/ani15182653 - 10 Sep 2025
Viewed by 482
Abstract
This review explores the potential of essential oils (EOs) as natural feed additives in aquaculture, highlighting their antimicrobial, antioxidant, and immunostimulatory properties, which contribute to disease prevention and improved fish resilience. EOs, derived from aromatic plants, offer a sustainable alternative to synthetic chemicals, [...] Read more.
This review explores the potential of essential oils (EOs) as natural feed additives in aquaculture, highlighting their antimicrobial, antioxidant, and immunostimulatory properties, which contribute to disease prevention and improved fish resilience. EOs, derived from aromatic plants, offer a sustainable alternative to synthetic chemicals, promoting benefits such as enhanced growth rates, feed efficiency, immune function, and reduced pathogen susceptibility. However, several challenges must be addressed to fully unravel their potential, including the optimization of dosages, effective delivery methods, and cost-efficiency. Techniques such as microencapsulation are emerging as promising solutions to improve EO stability and controlled release in aquatic feeds, though further research is needed to refine these approaches and evaluate their scalability. Additionally, there is a need for more research into the mechanisms through which EOs influence fish health, the interactions between active compounds, potential synergistic effects of EO mixtures, and their impact on the aquatic environment and microbiome. Addressing these challenges will ensure the effective and sustainable application of EOs in aquaculture, reducing reliance on synthetic chemicals while fostering a more resilient and eco-friendly industry. A key feature of this review is the systematic presentation of detailed, species-specific tables summarizing the current literature on the application of EOs and plant extracts in fish health management. Full article
20 pages, 1324 KB  
Review
Mycobacterium marinum Immune Evasion in Zebrafish
by Priyank Kumar, Joshua Cameron, Beatrice Saviola and Vishwanath Venketaraman
Pathogens 2025, 14(9), 908; https://doi.org/10.3390/pathogens14090908 - 10 Sep 2025
Viewed by 681
Abstract
Fish mycobacteriosis, a chronic progressive disease caused by nontuberculous mycobacteria (NTM), affects marine, brackish, and freshwater fish. Mycobacterium marinum (M. marinum), the most important of the NTM, infects fresh and marine water fish causing necrotizing granulomas and associated morbidity and mortality. [...] Read more.
Fish mycobacteriosis, a chronic progressive disease caused by nontuberculous mycobacteria (NTM), affects marine, brackish, and freshwater fish. Mycobacterium marinum (M. marinum), the most important of the NTM, infects fresh and marine water fish causing necrotizing granulomas and associated morbidity and mortality. M. marinum causes disease in zebrafish in a dose-dependent fashion. The M. marinum-induced disease in the zebrafish is associated with the development of necrotizing granulomas with abundant bacteria in the necrotic areas. Acute infection with high infectious doses of M. marinum infection in zebrafish was characterized by uncontrolled replication of the pathogen and death of all fish within 16 days, while chronic infections were marked by the formation of granulomas in different organs and longer survival in the range of 4–8 weeks. This review therefore synthesizes recent advances in our understanding of M. marinum’s infection of zebrafish, molecular pathogenesis, virulence mechanisms, and immune evasion strategies in zebrafish, while also highlighting the host immune effector responses and the virulence mechanisms of M. marinum. Full article
(This article belongs to the Special Issue Infectious Diseases in Aquatic Animals)
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25 pages, 2009 KB  
Article
Molecular Epidemiology, Antimicrobial Resistance, and Virulence Profiles of Staphylococcus aureus from Fish, Aquatic Environments, and Fish Handlers in Southeast Nigeria
by Uju Catherine Okafor, Onyinye Josephine Okorie-Kanu, Akwoba Joseph Ogugua, Chika Florence Ikeogu, Simeon Chibuko Okafor, Madubuike Umunna Anyanwu, Obichukwu Chisom Nwobi, Chidiebere Ohazuruike Anyaoha, Anthony Christian Mgbeahuruike, Lynda Onyinyechi Majesty-Alukagberie, Innocent Okwundu Nwankwo, Chukwunonso Francis Obi, Ejike Ekene Ugwuijem, Nkechi Harriet Ikenna-Ezeh, Ifeyinwa Riona Okosi, Onyemaechi Ugboh, George Okey Ezeifeka, Ekene Vivienne Ezenduka, Charles Odilichukwu R. Okpala and Edet Ekpenyong Udo
Microorganisms 2025, 13(9), 2059; https://doi.org/10.3390/microorganisms13092059 - 4 Sep 2025
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Abstract
Background: Staphylococcus aureus is a major zoonotic and foodborne pathogen with substantial One Health implications, yet its prevalence, resistance, and virulence potential within the aquaculture sector in Nigeria remains poorly characterized. Objectives: To supplement existing information, this current study investigated the [...] Read more.
Background: Staphylococcus aureus is a major zoonotic and foodborne pathogen with substantial One Health implications, yet its prevalence, resistance, and virulence potential within the aquaculture sector in Nigeria remains poorly characterized. Objectives: To supplement existing information, this current study investigated the prevalence, clonal distribution, antimicrobial resistance, and virulence gene profiles of S. aureus isolates from fish, fish water, and occupationally exposed fish handlers in Anambra State, Southeast Nigeria. Methods: A total of 607 samples—comprising 465 surface swabs from raw and processed fish, 36 fish water samples, and 106 nasal swabs from fish handlers—were processed using selective culture, biochemical tests, antimicrobial susceptibility testing, DNA microarray analysis, spa typing, and SCCmec typing. Results: S. aureus was recovered from 16.5% (100/607) of the samples. Fourteen (14%) isolates were methicillin-resistant (MRSA), harboring mecA and SCCmec types IV and V, with a combined MRSA prevalence of 2.3%. Multidrug resistance was observed in 52.2% of isolates (mean Multiple Antimicrobial Resistance index: 0.23), with 19 resistance genes spanning nine antimicrobial classes—including heavy metal and biocide resistance. Twenty-eight spa types across 13 clonal complexes (CCs) were identified, with CC1, CC5, and CC8 predominating. The detection of shared spa types between fish and handlers indicates potential cross-contamination. Detected virulence genes included those for accessory gene regulators (agrI-IV), Pantone–Valentine leucocidin (lukFS-PV), toxic shock syndrome (tsst-1), hemolysins (hla, hlb, hld/hlIII, hlgA), biofilm formation (icaA, icaD), immune evasion (chp, scn, sak), enterotoxins (sea, seb, sec, sed, egc, and others), exfoliative toxins (etA, etB), epidermal cells differentiation (edinA, edinB), and capsular types (cap5, cap8). Conclusions: This study reveals that the aquaculture sector in Southeast Nigeria serves as a significant reservoir of genetically diverse, multidrug-resistant S. aureus strains with robust virulence profiles. These findings highlight the necessity of integrated One Health surveillance and targeted interventions addressing antimicrobial use and hygiene practices within aquatic food systems. Full article
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