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This study examines the performance of an anaerobic fixed-film bioreactor under seasonal operating conditions prevailing in medium and small size Tequila factories: start-up, normal operation and particularly, during the restart-up after a long stop and starvation period. The proposed start-up procedure attained a stable biofilm in a rather short period (28 days) despite unbalanced COD/N/P ratio and the use of non-acclimated inoculum. The bioreactor was restarted-up after being shut down for 6 months during which the inoculum starved. Even when biofilm detachment and bioreactor clogging were detected at the very beginning of restart-up, results show that the bioreactor performed better as higher COD removal and methane yield were attained. CE-SSCP and Q-PCR analyses, conducted on the biofilm prokaryotic communities for each operating condition, confirmed that the high COD removal results after the …