Comparative In Vitro Analysis of Protozoan Genus Responses to Antiprotozoal Agents from Extracts, Albizia chinensis Leaf Flour, and Commercial Saponin

Faculty of Animal Sciences, Universitas Brawijaya, Malang 65145, Indonesia

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Abstract

The purpose of this study was to compare the in vitro antiprotozoal activity of Albizia chinensis leaf, Albizia chinensis extract, and commercial saponin against rumen protozoa. The process began with a six-hour incubation of Etawa crossbred goat rumen fluid under controlled anaerobic conditions. DNA was extracted and amplified using 18S rRNA primers, and amplicons were sequenced using Oxford Nanopore technology to identify the composition of prozoal genera. The analysis showed that the treatment inhibited several protozoan genera, with variations in spectrum and intensity. The strongest and broadest inhibition value was shown by commercial saponin and eliminated genera such as Entamoeba and Isotricha. Albizia chinensis leaf extract showed selective inhibition, while coarse flour provided a milder but broader effect, this could occur due to the synergism between saponins, flavonoids, and the fiber matrix. The results of the study inform that the level of processing will affect the specificity and potency of antiprotozoal activity. Albizia chinensis based products offer a sustainable plant-based alternative that can reduce protozoa and rumen methane production. This study recommends further research on bioactive compound profiles and in vivo validation to optimize the use of A. chinensis as a natural feed additive for ruminant microbial management.