The study investigated the effects of long-chain fatty acids (LCFA) on anaerobic sludge treating lipid-rich wastewater. It involved batch experiments with three sludge samples: two acclimated to lipids and one non-acclimated. The experiments aimed to observe the degradation of LCFA, specifically oleate and palmitate, by dosing them at concentrations ranging from 50 to 600 mg/L. Measurements of the cumulative methane production and the LCFA concentration, quantified as fat, oil, and grease (FOG) were performed. To ensure the sludge was free from other biodegradable substrates, part of the samples was pre-incubated without feed. The tests were conducted with both pre-incubated and non-incubated inoculum sludge. The findings revealed that oleate was degraded more efficiently than palmitate across all sludge samples, with a greater conversion rate to methane. Sludge samples acclimated to lipids showed a superior capacity to degrade LCFA compared to non-acclimated ones. It was noted that at concentrations above 400 mg/L, the conversion of LCFAs to intermediate compounds was inhibited, although this did not affect the subsequent methane production. The study concludes with a recommendation for sludge adaptation strategies to boost the efficiency of anaerobic wastewater treatment systems dealing with lipid-rich waste. The presence of LCFA-degrading bacteria families like Kosmotogaceae, Petrotogaceae, and Synergistaceae in the acclimated sludge samples underscores the adaptation and potential for improved degradation performance.