Identification of metabolic biomarkers and pathways associated with Ichthyophonus hoferi infection in White stumpnose (Rhabdosargus globiceps)

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Introduction Ichthyophonus hoferi is a cosmopolitan parasite infecting a wide range of fish species across marine, estuarine, brackish, and freshwater environments. Currently, diagnosis relies on destructive sampling methods, posing challenges for disease surveillance in aquaculture and wild populations. This study aimed to identify pre-clinical metabolic markers of I. hoferi infection in White stumpnose fish ( Rhabdosargus globiceps ) using non-lethal blood-based diagnostics. Methods An untargeted metabolomics approach employing comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry (GCxGC-TOFMS) was conducted on dried blood spot (DBS) samples. Multivariate (PCA, PLS-DA) and univariate analyses were used to evaluate metabolic differences between infected and control groups. Result A total of 655 metabolites were detected, with 98 showing significant variation between groups. Infected fish exhibited elevated levels of xenobiotics and altered carbohydrate metabolism, with key downregulated metabolites including D-mannose, arabinofuranose, and pento-3-ulose. Several fatty acids, such as oleic acid and pentadecanoic acid, were also significantly altered. Pathway analysis revealed that propanoate metabolism, galactose metabolism, inositol phosphate metabolism, and the TCA cycle were significantly impacted by I. hoferi infection, suggesting disruption in energy production and anti-inflammatory pathways. Among the top discriminatory metabolites, D-mannose was notably downregulated and identified as a potential biomarker for I. hoferi infection. Discussion This study demonstrates that metabolomics, in combination with DBS sample collection, offers a promising non-destructive approach for detecting parasitic infections in fish. The findings contribute to the development of diagnostic tools for early detection and monitoring of I. hoferi infection, with implications for fish health management and conservation.

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