Search Results (76)

Background/Objectives: The overuse and misuse of antibiotics has contributed significatively to the growing problem of the emergence and spread of antibiotic resistance genes among bacteria, posing a serious global challenge to the treatment of bacterial infectious diseases. For these reasons, there is a current and growing interest in the development of effective alternative or complementary strategies to antibiotic therapy for the prevention of fish diseases, which are mainly based on the use of probiotics—in particular, those belonging to the Lactic Acid Bacteria (LAB) group. In this context, the aim of the present study was to characterise, evaluate the genetic diversity and assess the safety of candidate probiotic LAB strains for aquaculture isolated from faeces and intestines of European hakes (Merluccius merluccius, L.) caught in the Northeast Atlantic Ocean (Ireland). Methods: The direct antimicrobial activity of the LAB isolates was tested by the Stab-On-Agar method against key ichthyopathogens. Subsequently, their taxonomic classification and genetic diversity were determined by 16SrDNA sequencing and Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR), respectively. To ensure the in vitro safety of the LAB isolates, their biofilm-forming ability was assessed by a microtiter plate assay; their sensitivity to major antibiotics used in aquaculture, human and veterinary medicine by a broth microdilution method and their haemolytic and gelatinase activity by microbiological assays. Results: All LAB isolates were biofilm producers and susceptible to chloramphenicol, oxytetracycline, flumequine and amoxicillin. A total of 30 isolates (85.7%) were resistant to at least one of the tested antibiotics. None of the 35 LAB isolates showed haemolytic or proteolytic activity. Conclusions: Among the isolated strains, five LAB strains exhibiting the highest antimicrobial activity against aquaculture-relevant ichthyopathogens, taxonomically identified as Streptococcus salivarius, Enterococcus avium and Latilactobacillus sakei, were selected for further characterisation as potential probiotic candidates to promote sustainable aquaculture. To our knowledge, this is the first study to report that hake intestines and faeces represent viable ecological niches for the isolation of LAB strains with antimicrobial activity. Full article

Ornamental fish represent a significant aquaculture sector with notable economic value, yet their contribution to antibiotic residues and resistance remains underrecognized. This review synthesizes evidence on widespread and often unregulated antibiotic use—including tetracyclines and fluoroquinolones—in ornamental fish production, transportation, and retail, primarily targeting bacterial diseases such as aeromonosis and vibriosis. Pathogenic microorganisms including Edwardsiella, Flavobacterium, and Shewanella spp. cause diseases like hemorrhagic septicemia, fin rot, skin ulcers, and exophthalmia, impairing fish health and marketability. Prophylactic and therapeutic antibiotic applications elevate antibiotic residues in fish tissues and carriage water, thereby selecting for antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs). These resistant elements pose significant risks to fish health, human exposure via direct contact and bioaerosols, and environmental health through contamination pathways. We emphasize the urgent need for a holistic One Health approach, involving enhanced surveillance, stringent regulatory oversight, and adoption of alternative antimicrobial strategies, such as probiotics and advanced water treatments. Coordinated global actions are crucial to effectively mitigate antibiotic resistance within the ornamental fish industry, ensuring sustainable production, safeguarding public health, and protecting environmental integrity. Full article

In humans, certain functions such as language and spatial attention are lateralised, meaning they are predominantly or exclusively performed by one hemisphere. Intriguingly, a significant portion of individuals exhibit a mirrored pattern of asymmetry, which has been attributed to genes, environmental influences, or other factors. As lateralisation occurs across all major groups of vertebrates, literature in other species might provide valuable insights into these mechanisms. We reviewed current knowledge on the genetic and environmental factors underlying individual variability in lateralisation in non-human vertebrates. Evidence of a genetic basis for the direction, strength of asymmetries, or both has been reported in about a dozen species of fish and mammals. Nevertheless, a careful examination revealed that none of these studies can definitively exclude the influence of non-genetic factors. On the other hand, studies from the past decade have suggested that environmental factors can shape both the direction and strength of lateralisation in adaptive ways, aligning the individual’s phenotype with local environmental conditions. Overall, this review supports the potential role of both genetic differences and environmentally driven plasticity in shaping lateralisation variance while highlighting literature gaps that prevent a precise disentanglement of the roles of these factors. Full article

Background/Objectives: This study explores the metabolic adaptations and quality differences between wild and farmed gilthead seabream (Sparus aurata), with a particular focus on lipid composition and metabolite profiles. These differences are examined in the context of varying environmental conditions, feeding regimes, and post-harvest processes. High-resolution magic-angle-spinning nuclear magnetic resonance (HR-MAS NMR) spectroscopy was employed to perform the metabolomic analysis. Results: Farmed seabream exhibited higher lipid content and PUFA levels (e.g., DHA and EPA) due to aquaculture diets, while wild seabream showed lower lipid concentrations and elevated levels of polar metabolites. Metabolic trade-offs in wild seabream reflected greater physical activity and environmental adaptation. The K-value indicated faster spoilage in farmed seabream, particularly from Greece, linked to handling conditions. HR-MAS provided precise, reproducible results, allowing direct quantification of key metabolites without altering sample integrity. Methods: HR-MAS NMR was employed to analyze muscle tissue from wild and farmed seabream (produced in Spain and imported from Greece), providing high-resolution spectra without requiring sample extraction. Metabolite quantification included polyunsaturated fatty acids (PUFAs), creatine, taurine, lactate, and trimethylamine N-oxide (TMAO). Freshness was monitored using the K-value index, calculated from ATP derivative levels in samples stored at 4 °C. Conclusions: The study highlights the influence of diet and environment on the metabolic profiles of seabream. HR-MAS NMR emerges as a robust method for metabolomic studies and freshness assessment. Findings emphasize the potential for dietary adjustments to optimize aquaculture practices and fish quality while underscoring the importance of sustainable production strategies. Further research into lipid metabolism genes and environmental factors is recommended to deepen understanding of these adaptations. Full article

As a fundamental taxonomic group within vertebrates, fish represent an invaluable resource for investigating the mechanisms underlying sex determination and differentiation owing to their extensive geographical distribution and rich biodiversity. Within this biological cohort, the processes of sex determination and differentiation are intricately governed by both genetic factors and the complex interplay of environmental cues. While variations in external environmental factors, particularly temperature, can exert a modulatory influence on sex differentiation in fish to a limited degree, genetic factors remain the primary determinants of sexual traits. Hermaphroditic fish display three distinct types of sexual transitions: protandry (male to female), protogyny (female-to-male), bidirectional sex change (both directions serially). These fish, characterized by their unique reproductive strategies and sexual plasticity, serve as exemplary natural models for elucidating the mechanisms of sex differentiation and sexual transitions in fish. The present review delves into the histological dynamics during gonadal development across three types of sequential hermaphroditic fish, meticulously delineating the pivotal characteristics at each stage, from the inception of primordial gonads to sexual specialization. Furthermore, it examines the regulatory genes and associated signaling pathways that orchestrate sex determination and differentiation. By systematically synthesizing these research advancements, this paper endeavors to offer a comprehensive and profound insight into the intricate mechanisms governing sex differentiation in sequential hermaphroditic fish. Full article

Vitellogenins (Vtgs) are key yolk precursor proteins in fish, serving as critical indicators of gonadal maturation in females and reliable biomarkers for detecting xeno-oestrogenic pollution, particularly through their expression in juveniles or males. The vtg gene family comprises multiple subtypes that are species-specific, necessitating precise characterisation and quantification for effective use as biomarkers in studies on estrogenic endocrine-disrupting chemicals (EEDCs). In this study, we successfully cloned and characterised the full-length cDNAs of three vtg subtypes (vtgAa, vtgAb, and vtgC) from Scatophagus argus. Differential expression analysis revealed that vtgAb exhibited the highest responsiveness to 17α-ethynylestradiol (EE2) exposure, with a 3-fold increase in vivo at 10.0 μg/g EE2 and a 30-fold increase in vitro at 10⁻⁷ mol/L EE2. The expression patterns were dose- and time-dependent, with peak expression observed 72 h post-exposure. While in vivo assays indicated moderate upregulation, in vitro experiments demonstrated significantly higher expression, attributed to direct hepatocyte interaction with EE2. These findings confirm vtgAb as the most responsive subtype to oestrogen exposure in S. argus and highlight the species’ tolerance to EE2, as compared to more sensitive species like Danio rerio. This study shows the evolutionary conservation of vtg transcripts across teleost species and reinforces the importance of subtype-specific characterisation to advance their application as biomarkers for EEDCs, with significant implications for environmental monitoring and pollution regulation. Full article

The Silway River has historically failed to meet safe fecal coliform levels due to improper waste disposal. The river mouth is located in General Santos City, the tuna capital of the Philippines and a leading producer of hogs, cattle, and poultry. The buildup of contaminants due to direct discharge of waste from chicken farms and existing water quality conditions has led to higher fecal matter in the Silway River. While there were technical reports in the early 2000s about poultry farming, this is the first study where fecal coliform from poultry farming was detected in the Silway River using highly sensitive protocols like qPCR. This study characterized the effect of flow velocity and physicochemical water quality parameters on chicken fecal contamination. Gene markers such as Ckmito and ND5-CD were used to detect and quantify poultry manure contamination through microbial source tracking (MST) and environmental DNA (eDNA) profiling. The results of this study showed the presence of chicken fecal bacteria in all stations along the Silway River. The results revealed that normal levels of water quality parameters such as temperature, pH, and high TSS concentrations create favorable conditions for chicken fecal coliforms to thrive. Multiple regression analysis showed that flow velocity and DO significantly affect chicken fecal contamination. A lower cycle threshold (Ct) value indicated higher concentration of the marker ND5-CD, which means higher fecal contamination. It was found that there was an inverse relationship between the Ct value and both velocity (R² = 0.55, p = 0.01) and DO (R² = 0.98, p = 0.2), suggesting that low flow velocity and low DO can lead to higher fecal contamination. Findings of fecal contamination could negatively impact water resources, the health of nearby residents, and surrounding farms and industries, as well as the health and growth of fish. Full article

The problem of marine noise pollution has a long history. Strong noise (>120 dB re 1 µPa) will affects the growth, development, physiological responses, and behaviors of fish, and also can induce the stress response, posing a mortal threat. Although many studies have reported that underwater noise may affect the survival of fish by disturbing their nervous system and endocrine system, the underlying causes of death due to noise stimulation remain unknown. Therefore, in this study, we used the underwater noise stress models to conduct underwater strong noise (50–125 dB re 1 µPa, 10–22,000 Hz) stress experiments on small yellow croaker for 10 min (short-term noise stress) and 6 days (long-term noise stress). A total of 150 fishes (body weight: 40–60 g; body length: 12–14 cm) were used in this study. Omics (metabolomics and transcriptomics) studies and quantitative analyses of important genes (HPA (hypothalamic–pituitary–adrenal)-axis functional genes) were performed to reveal genetic and metabolic changes in the important tissues associated with the HPA axis (brain, heart, and adrenal gland). Finally, we found that the strong noise pollution can significantly interfere with the expression of HPA-axis functional genes (including corticotropin releasing hormone (CRH), corticotropin releasing hormone receptor 2 (CRHR2), and arginine vasotocin (AVT)), and long-term stimulation can further induce metabolic disorders of the functional tissues (brain, heart, and adrenal gland), posing a lethal threat. Meanwhile, we also found that there were two kinds of death processes, direct death and chronic death, and both were closely related to the duration of stimulation and the regulation of the HPA axis. Full article

Difenoconazole (DIF) is a fungicide used to control various fungi. It is absorbed on the surface of different plants and contributes significantly to increased crop production. However, DIF is reported to exhibit toxicity to fungi and to aquatic plants, fish, and mammals, including humans, causing adverse effects. However, research on the impact of DIF on the mammary epithelial cells of herbivorous bovines is limited. DIF-induced damage and accumulation in the mammary glands can have direct and indirect effects on humans. Therefore, we investigated the effects and mechanisms of DIF toxicity in MAC-T cells. The current study revealed that DIF reduces cell viability and proliferation while triggering apoptotic cell death through the upregulation of pro-apoptotic proteins, including cleaved caspase 3 and Bcl-2-associated X protein (BAX), and the downregulation of leukemia type 2 (BCL-2). DIF also induced endoplasmic reticulum (ER) stress by increasing the expression of genes or proteins of Bip/GRP78, protein disulfide isomerase (PDI), activating transcription factor 4 (ATF4), C/EBP homologous protein (CHOP), and endoplasmic reticulum oxidoreductase 1 Alpha (ERO1-Lα). We demonstrated that DIF induces mitochondria-mediated apoptosis in MAC-T cells by activating ER stress pathways. This cellular damage resulted in a significant increase in the expression of inflammatory response genes and proteins, including cyclooxygenase 2 (COX2), transforming growth factor beta 3 (TGFB3), CCAAT enhancer binding protein delta (CEBPD), and iNOS, in DIF-treated groups. In addition, spheroid formation by MAC-T cells was suppressed by DIF treatment. Our findings suggest that DIF exposure in dairy cows may harm mammary gland function and health and may indirectly affect human consumption of milk. Full article

European sea bass (Dicentrarchus labrax) is one of the most significant species farmed in the Mediterranean, yet a very perishable product. Its quality deteriorates rapidly as a result of three mechanisms: microbial activity, chemical oxidation, and enzymatic degradation. Microbial spoilage is the mechanism that contributes most to the quality deterioration of fresh and non-processed fish. To this end, our study aims to identify for the first time the combined effect of aquatic environment and harvest method on the composition and trajectory at storage at 0 °C of the European sea bass skin microbiome. Sampling was performed in two commercial fish farms in Western (WG) and Central Greece (CG) where fish were harvested using different methods: direct immersion in ice water or a mixture of slurry ice; application of electro-stunning prior to immersion in ice water. Samples were collected on harvest day and one week post-harvest. To profile the bacterial communities in the fish skin, 16S ribosomal RNA gene sequencing was used. The results and the following analyses indicated that the aquatic environment shaped the original composition of the skin microbiome, with 815 ASVs identified in the WG farm as opposed to 362 ASVs in the CG farm. Moreover, Pseudomonas and Pseudoalteromonas dominated the skin microbiome in the WG farm, unlike the CG farm where Shewanella and Psychrobacter were the dominant genera. All these genera contain species such as Shewanella putrefaciens, Pseudomonas aeruginosa, Pseudoalteromonas spp., and Psychrobacter sp., all of which have been implicated in the deterioration and spoilage of the final product. The different harvest methods drove variations in the microbiome already shaped by the aquatic environment, with electro-stunning favoring more diversity in the skin microbiome. The aquatic environment in combination with the harvest method appeared to determine the skin microbiome trajectory at storage at 0 °C. Although Shewanella had dominated the skin microbiome in all samples one week post-harvest, the diversity and the relative abundance of genera were strongly influenced by the aquatic environment and the harvest method. This study sheds light on the hierarchy of the factors shaping the fish skin microbiome and their importance for controlling post-harvest quality of fresh fish. Full article

Lead (Pb) is a well-known neurotoxin with established adverse effects on the neurological functions of children and younger adults, including motor, learning, and memory abilities. However, its potential impact on older adults has received less attention. Using the zebrafish model, our study aims to characterize the dose–response relationship between environmentally relevant Pb exposure levels and their effects on changes in behavior and transcriptomics during the geriatric periods. We exposed two-year-old zebrafish to waterborne lead acetate (1, 10, 100, 1000, or 10,000 µg/L) or a vehicle (DMSO) for 5 days. While lower concentrations (1–100 µg/L) reflect environmentally relevant Pb levels, higher concentrations (1000–10,000 µg/L) were included to assess acute toxicity under extreme exposure scenarios. We conducted adult behavior assessment to evaluate the locomotor activity following exposure. The same individual fish were subsequently sacrificed for brain dissection after a day of recovery in the aquatic system. RNA extraction and sequencing were then performed to evaluate the Pb-induced transcriptomic changes. Higher (1000–10,000 ug/L) Pb levels induced hyperactive locomotor patterns in aged zebrafish, while lower (10–100 ug/L) Pb levels resulted in the lowest locomotor activity compared to the control group. Exposure to 100 µg/L led to the highest number of differentially expressed genes (DEGs), while 10,000 µg/L induced larger fold changes in both directions. The neurological pathways impacted by Pb exposure include functions related to neurotransmission, such as cytoskeletal regulation and synaptogenesis, and oxidative stress response, such as mitochondrial dysfunction and downregulation of heat shock protein genes. These findings emphasize a U-shape dose–response relationship with Pb concentrations in locomotor activity and transcriptomic changes in the aging brain. Full article

The rise of carbapenem-resistant Klebsiella pneumoniae (CRKP) presents a significant global challenge in clinical and healthcare settings, severely limiting treatment options. This study aimed to utilize a bacteriophage as an alternative therapy against carbapenem-resistant K. pneumoniae. A novel lytic N4-like Klebsiella phage, vB_kpnP_KPYAP-1 (KPYAP-1), was isolated from sewage. It demonstrated efficacy against the K62 serotype polysaccharide capsule of bla_OXA-48-producing K. pneumoniae. KPYAP-1 forms small, clear plaques, has a latent period of 20 min, and reaches a growth plateau at 35 min, with a burst size of 473 plaque-forming units (PFUs) per infected cell. Phylogenetic analysis places KPYAP-1 in the Schitoviridae family, Enquatrovirinae subfamily, and Kaypoctavirus genus. KPYAP-1 employs an N4-like direct terminal repeat mechanism for genome packaging and encodes a large virion-encapsulated RNA polymerase. It lacks integrase or repressor genes, antibiotic resistance genes, bacterial virulence factors, and toxins, ensuring its safety for therapeutic use. Comparative genome analysis revealed that the KPYAP-1 genome is most similar to the KP8 genome, yet differs in tail fiber protein, indicating variations in host recognition. In a zebrafish infection model, KPYAP-1 significantly improved the survival rate of infected fish by 92% at a multiplicity of infection (MOI) of 10, demonstrating its potential for in vivo treatment. These results highlight KPYAP-1 as a promising candidate for developing phage-based therapies targeting carbapenemase-producing K. pneumoniae. Full article

Owing to the presence of drugs targeting adrenergic receptors in aquatic ecosystems, considerable attention has been directed towards their environmental distribution and fate in recent decades. However, their potential impacts on non-target aquatic organisms, particularly fish, have received relatively limited investigation. In this study, moxisylyte (MOX) and propranolol (PRO) were selected as representatives of α- or β-adrenergic receptor antagonist, respectively, and we assessed their effects on the early life stages of zebrafish, especially on the nervous and cardiovascular systems. Although both compounds exhibited marginal effects on zebrafish survival, hatching and gross abnormality following exposure to concentrations ranging from 1 to 625 μg/L, they adversely affected the development of cardiovascular and nervous systems, but through different mechanisms of action, as evidenced by variations in gene transcriptional responses and enzyme activities. Notably, cardiovascular responses appear promising for use as potential biomarkers for exposure to drugs targeting adrenergic receptors. This study enhances our understanding of the ecotoxicological risks posed by α- and β-blockers in fish. Nonetheless, further investigation is needed to elucidate the precise mechanisms underlying the impacts of drugs targeting adrenergic receptors due to our limited knowledge of the physiological functions of the adrenergic system in fish. Full article

E3 ubiquitin ligases, key components of the ubiquitin proteasome system, orchestrate protein degradation through ubiquitylation and profoundly impact cellular biology. Small HERC E3 ligases (HERC3-6) have diverse functions in mammals, including roles in spermatogenesis, protein degradation, and immunity. Until now, only mammals’ HERC3, HERC5, and HERC6 are known to participate in immune responses, with major involvement in the antiviral response. Interestingly, an exclusive HERC7 has been characterized in fish showing great molecular conservation and antiviral roles. Thus, this study identifies and characterizes the herc7 gene in the European sea bass teleost. The European sea bass herc7 gene and the putative protein show good conservation of the promoter binding sites for interferons and the RCC1 and HECT domains characteristic of HERC proteins, respectively. The phylogenetic analysis shows a unique cluster with the fish-exclusive HERC7 orthologues. During ontogeny, the herc7 gene is expressed from 3 days post-fertilization onwards, being constitutively and widely distributed in adult tissues. In vitro, stimulated leucocytes up-regulate the herc7 gene in response to mitogens and viruses, pointing to a role in the immune response. Furthermore, sea bass herc7 expression is related to the interferon response intensity and viral load in different tissues upon in vivo infection with red-grouper betanodavirus (RGNNV), suggesting the potential involvement of fish HERC7 in ISGylation-based antiviral activity, similarly to mammalian HERC5. This study broadens the understanding of small HERC proteins in fish species and highlights HERC7 as a potential contributor to the immune response in European sea bass, with implications for antiviral defense mechanisms. Future research is needed to unravel the precise actions and functions of HERC7 in teleost fish immunity, providing insights into direct antiviral activity and viral evasion. Full article

The most significant genetic influence on eye color pigmentation is attributed to the intronic SNP rs12913832 in the HERC2 gene, which interacts with the promoter region of the contiguous OCA2 gene. This interaction, through the formation of a chromatin loop, modulates the transcriptional activity of OCA2, directly affecting eye color pigmentation. Recent advancements in technology have elucidated the precise spatial organization of the genome within the cell nucleus, with chromatin architecture playing a pivotal role in regulating various genome functions. In this study, we investigated the organization of the chromatin close to the HERC2/OCA2 locus in human lymphocyte nuclei using fluorescence in situ hybridization (FISH) and high-throughput chromosome conformation capture (Hi-C) data. The 3 Mb of genomic DNA that belonged to the chromosomal region 15q12-q13.1 revealed the presence of three contiguous chromatin loops, which exhibited a different level of compaction depending on the presence of the A or G allele in the SNP rs12913832. Moreover, the analysis of the genomic organization of the genes has demonstrated that this chromosomal region is evolutionarily highly conserved, as evidenced by the analysis of syntenic regions in species from other Vertebrate classes. Thus, the role of rs12913832 variant is relevant not only in determining the transcriptional activation of the OCA2 gene but also in the chromatin compaction of a larger region, underscoring the critical role of chromatin organization in the proper regulation of the involved genes. It is crucial to consider the broader implications of this finding, especially regarding the potential regulatory role of similar polymorphisms located within intronic regions, which do not influence the same gene by modulating the splicing process, but they regulate the expression of adjacent genes. Therefore, caution should be exercised when utilizing whole-exome sequencing for diagnostic purposes, as intron sequences may provide valuable gene regulation information on the region where they reside. Thus, future research efforts should also be directed towards gaining a deeper understanding of the precise mechanisms underlying the role and mode of action of intronic SNPs in chromatin loop organization and transcriptional regulation. Full article