Anaerobic digestion (AD) is a process resulting from the anaerobic metabolism of specific microorganisms that produce an eco-friendly type of energy and a stabilized soil fertilizer. We described the microbial communities and their changes in three depths of BioKöhler® biodigester, fed with cattle manure for 18 days, under anaerobic incubation at the psychrophilic temperature range (~ 20 °C). During the experiment, the maximum methane content in the raw biogas was 79.9%. Non-metric

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... nal scaling (MDS) showed significant differences among microbial communities in the bottom, medium, and upper depths. Considering all the periods of incubation, the microbial communities changed until the eighth day, and they remained stable from eighth to seventeenth days. Bacteroidetes, Firmicutes, and Synergistetes were the most abundant phyla in samples, representing approximately 41% of the total OTUs. The relative abundance of the phyla Euryarchaeota, Actinobacteria, Firmicutes, and Verrucomicrobia changed from bottom to medium sampling points. Moreover, Crenarchaeota differed in frequencies from medium to upper, and Acidobacteria from bottom to upper samples. Lentisphaerae, Chloroflexi, and LD1 were different solely at the bottom, whereas OP9 and Tenericutes only in the medium. Psychrophilic AD performed in this work removed pathogens like Salmonella and Escherichia, as observed at the digestate analyzed. This type of treatment of raw manure besides producing eco-friendly energy efficiently also generates a stabilized and safe biomass that can be used as fertilizer in soils.