Mixed Culture Optimization for Marigold Flower Ensilage via Experimental Design and Response Surface Methodology

Endogenous microorganisms isolated from the marigold flower (Tagetes erecta) were studied to understand the events taking place during its ensilage. Studies of the cellulase enzymatic activity and the ensilage process were undertaken. In both studies, the use of approximate second-order models and multiple lineal regression, within the context of an experimental mixture design using the response surface methodology as optimization strategy, determined that the microorganisms Flavobacterium IIb, Acinetobacter anitratus, and Rhizopus nigricans are the most significant in marigold flower ensilage and exhibit high cellulase activity. A mixed culture comprised of 9.8% Flavobacterium IIb, 41% A. anitratus, and 49.2% R. nigricans used during ensilage resulted in an increased yield of total xanthophylls extracted of 24.94 g/kg of dry weight compared with 12.92 for the uninoculated control ensilage. Keywords: Marigold flower; ensilage; starter inoculum; mixed culture