Immunological and molecular evaluation of zoonotic metacercarial infection in freshwater fish: A cross-sectional analysis

Improperly cooked fish, carrying active metacercariae (MCs), can pose a significant risk for transmitting fish-borne zoonotic trematodes (FBZTs) to human consumers. This study aimed to enhance our understanding of FBZTs by conducting a comprehensive cross-sectional analysis involving various fish species, such as Nile tilapia (Oreochromis niloticus), African catfish (Clarias gariepinus), and red-belly tilapia (Tilapia zillii). These fish specimens were collected from distinct Egyptian governorates, specifically Giza, Kafr al-Shaykh, and Fayoum. The recovered flukes from experimentally infected domestic pigeons were identified as Prohemistomum vivax, Haplorchis pumilio, and Pygidiopsis genata based on morphological features. Furthermore, the identity of the retrieved adult flukes was confirmed using three species-specific primers for PCR amplification and sequencing analysis of the ITS rDNA region and have been deposited in GenBank with the following accession numbers: P. vivax (OR291421.1 and OR291422.1), P. genata (OP099561.1), and H. pumilio (OM439581.1-OP090510.1). Quantitative real-time PCR targeting the immunological genes Tumor Necrosis Factor-alpha (TNF-alpha) and Interleukin-1 (IL-1Β) was employed to compare the cellular immune response between infected with EMCs and uninfected O. niloticus. The results indicated a significant increase in TNF- and IL-1Β levels in FBZTs-infected vs un-infected fishes. Importantly, the presence of adult flukes and EMCs led to substantial histological alterations in both experimentally infected pigeons and naturally infected fish tissues. These changes included the necrosis of fish muscle bundles and a pronounced inflammatory reaction with muscular necrosis in the digestive tracts of experimentally infected pigeons. [Display omitted] •In Egypt, Cross-sectional analysis of fish-borne zoonotic trematodes (FBZTs) in freshwater fishes in three different geographical areas is an alarming threat.•FBZTs confirmed with molecular characterization the threat of being heavily infected with three trematodes P. vivax, H. pumilio, and P. genata parasites.•Assessed gene expression and histopathology in these infected fish, finding that elevated cytokine levels, specifically TNF-α and IL-1Β β, correlated with trematode infections.•FBZT infections were found to have detrimental effects on fish health and could potentially pose risks to human health.•There is an urgent need to improve public health, emphasizing the need for continuous