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1st Int. Transborder Conf. of the Timor Island: Timor %u2013 Science without borderDili, 7-8 May 202527Reducing The Anti-Nutritional Properties of Chromolaena OdorataThrough Biofermentation For Use As A Potential Low-Cost Protein Source For Non-RuminantsM. L. Mullik1*, H. Z. Kotta1, I. Benu1, P. De Deus2, Acacio C. Amaral2, and N. Da Costa21Fakultas Peternakan, Perikanan dan Kelautan, Universitas Nusa Cendana, Indonesia2Instituto Politecnico de Betano, Same, Timor-Leste.*Corresponding author: marthenmullik@staf.undana.ac.idAbstractThe main constraint in utilizing Chromolaena odorata as an alternative low-cost protein source for livestock is its anti-nutritional properties, including anti-trypsin, tannins, oxalate, nitrate, phytate, and saponin. A series of studies conducted by researchers at Universitas Nusa Cendana in West Timor, Indonesia, found that biofermentation reduced these anti-nutritional compounds, making the plant safer for use as ruminant feed. However, those studies used whole aerial parts (leaves and stems) of C. odorata, resulting in a high fiber content in the meal. Therefore, it is unsuitable for non-ruminants, which are intolerant to high fiber levels. The present study aimed to assess the effects of biofermenting C. odorata leaves on crude fiber and the concentrations of anti-trypsin, tannins, nitrate, saponin, and oxalate in the fermented products. A controlled experiment was conducted using a completely randomized design with four treatments: nonbiofermented Chromolaena leaves (NBL, control), biofermented freshly harvested leaves (FBL), biofermented air-dried leaves (DBL), and biofermented post-water-soaked leaves (SBL). Each treatment had five replicates. A total of 25 plastic containers (5 kg capacity each) were used as silos. After harvesting and separating the leaves from the stems, the leaves were divided into four portions and assigned to the treatments. Biofermentation followed silage-making principles, using 100 mL of commercial mixed effective microorganisms (EM4 for feed) per 100 kg of fresh leaves (vol/wt). The fermentation process lasted seven days. Samples from each silo were collected and analyzed innutrition laboratory. The data were subjected to analysis of variance. Results showed that anti-trypsin content in the control group (4.06 mg/g) was significantly higher than in the treated groups (11.5%u201330.7 mg/g), with the lowest concentration (11.53 mg/g) found in the SBL treatment. Total tannin content followed a similar trend, with the highest value (3.74%) in the control group and the lowest (0.02%) in SBL. Similar trends were observed for saponin, phytate, and oxalate levels. It can be concluded that the concentrations of anti-trypsin, tannins, saponin, phytate, and oxalate were significantly reduced through biofermentation, with the lowest values observed in the post-water-soaked (SBL) treatment group.Keywords: chromolaena leaf, antri-nutrients, biofermentation, protein source, nonruminants.