Integrated evaluation of pineapple peel juice powder as a potential agro-waste-derived flukicidal preparation against Paramphistomum spp.

Paramphistomiasis is a neglected trematode infection affecting tropical livestock and remains inadequately addressed due to limited therapeutic options and underdeveloped evaluation frameworks. This study conducted an integrated in vitro-in vivo evaluation of the flukicidal activity and field efficacy of pineapple peel juice powder (PPJP), a whole-matrix agro-waste-derived peparation, against Paramphistomum spp. In vitro activity was assesed using adult motility assays (AMA) across a concentration gradient, and the 50% lethal concentration (LC₅₀) was estimated using a four-parameter logistic model. Field efficacy was evaluated in naturally infected Bali cattle using the faecal egg count reduction test (FECRT), while generalised estimating equations (GEE) were employed to model time-dependent outcomes in both assays. PPJP induced 100% mortality in adult flukes within 60 min, with an apparent LC₅₀ of 16.56 ng/mL observed at 90 min. In cattle, PPJP reduced faecal egg counts by 85.2% on day 7 and 100% on day 14 post-treatment. Scanning electron microscopy (SEM) demonstrated extensive tegumental collapse, deep cracking, and papillary loss, while histopathological examination confirmed marked tegumental sloughing and degeneration of sub-tegumental musculature. Preliminary LC-HRMS-based metabolite profiling identified diverse metabolite classes, providing chemical context for the observed biological effects. Overall PPJP exhibits rapid-onset flukicidal activity in vitro and measurable field efficacy against Paramphistomum spp, at selected dose, supporting further development of pineapple peel waste as a functional veterinary preparation aligned with agro-waste valorisation and sustainable livestock health strategies.