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Evaluation of hygienic properties of residues after anaerobic digestion (digestate) of dairy cow manure estimated from biological analysis

岩崎 匡洋 帯広畜産大学

2023.01.27

概要

To determine the effects of anaerobic digestion treatment on the survival of weed seeds in dairy cow manure, weed seeds (Rumex obtusifolius L.) were mixed into dairy cow manure, and the seed germination rates and percentage of dormant seeds after mesophilic (35 °C) and thermophilic (55 °C) anaerobic digestion processes were investigated in Chapter 1. The survival rates of Rumex obtusifolius L. seeds heat-treated at 35 °C and 55 °C were 75.5% and 0%, respectively, compared to 81.6% and 0% in anaerobic digestion processes at the same temperatures. The survival rates at 35 °C treatment were similar between the anaerobic digestion and heat treatments, but the percentage of seeds in primary and secondary dormancy was higher in the anaerobic digestion treatment compared to heat treatment. Although digestate have a high value as liquid fertilizer, mesophilic digestate should be treated with caution owing to the presence of dormant weed seeds.

In Chapter 2, the elimination of pathogenic bacteria in livestock manure during anaerobic diegstion in a full-scale biogas plant was investigated using livestock manure as a raw material. Manure was collected from reception pits, digester tanks, and digestate storage tanks of four biogas plants with mesophilic (38 °C) and thermophilic (55 °C) fermenters currently in operation in Hokkaido, Japan. Coli-aerogenes and Enterococcus in manure were cultured to analyze the survival rates after the digestion process. As a result, no Coli-aerogenes and Enterococcus were detected in either manure in the fermenter or digestate after anaerobic digestion in the thermophilic biogas plant. In the mesophilic biogas plant, although the number of Coli-aerogenes and Enterococcus decreased with digestion, Coli-aerogenes and Enterococcus in manure were found in the fermenter and digestate after anaerobic digestion. The mean decimation reduction time (T90) of Coli-aerogenes and Enterococcus in mesophilic digestion were 13.3 and 16.7 days, respectively. extended-spectrum β-lactamase (ESBL)-producing bacteria  (E. coli, Large quantities of antimicrobial agents have been used in dairy farming to treat and prevent livestock diseases. There has been a global problem of the resulting emergence of antimicrobial- resistant bacteria in livestock manure. Cefazolin (CEZ) is the antibiotic most commonly used in treating dairy mastitis, and CEZ-resistant (CEZ-R) bacteria are detected in manure of dairy cows administered with the antimicrobial. The study in Chapter 3, the effects of thermophilic anaerobic digestion on the survival of CEZ-R and CEZ-susceptible (CEZ-S) bacteria and substrate-specific Klebsiella/Enterobacter/Citrobacter, Proteus, Pseudomonas, and Acinetobacter) in dairy cow manure was examined. As a result, the number of CEZ-R bacteria was reduced to 0.93% by thermophilic anaerobic digestion, including a decrease in the number of E. coli, other coliforms, and other bacteria to below the limit of detection, 0.27%, and 1.05%, respectively. In contrast, there was a slight increase in ESBL-producing Pseudomonas. These results suggested that the risk of spreading cephazolin- resistant bacteria was reduced in the thermophilic anaerobic digestion process of livestock manure, but some antimicrobial-resistant bacteria remained.

The study in Chapter 4 also focused on the analysis of CEZ-R Pseudomonas spp. because there was a slight increase in ESBL-producing Pseudomonas spp. during thermophilic anaerobic digestion. Due to the mixture of pathogenic and plant growth-promoting species in Pseudomonas spp., P. aeruginosa, P. fluorescens, and other pseudomonads were classified and quantitatively analyzed using commercial PASA medium and newly prepared CAP medium (commercial Cetrimide medium to which L-arginine and phenol red were added to classify species according to differences in color development).

The evidence for these classifications was performed by PCR confirmation. Pseudomonas spp. decreased after both mesophilic and thermophilic anaerobic digestion, indicating that anaerobic digestion at thermophilic was more effective than at mesophilic during the anaerobic digestion process. P. aeruginosa was not detected in either manure collected or digestate after the mesophilic and thermophilic anaerobic digestion process. In contrast, CEZ-R Pseudomonas in mesophilic digestate increased relative to manure before the digestion process. P. fluorescens, which was present in manure and all CEZ-R, was no longer detected after mesophilic or thermophilic anaerobic digestion. Fluorescent pseudomonads, other than P. aeruginosa and P. fluorescens, increased after the anaerobic digestion process, and most of them were CEZ-S. In addition, an increase in the mesophilic anaerobic digestion process was observed in CEZ-R Pseudomonas spp. other than fluorescent pseudomonads. The percentage of bacterial counts suggested that most CEZ-R Pseudomonas spp. increased by the mesophilic anaerobic digestion process were other than fluorescent pseudomonads. Thus, due to the increase of CEZ-R Pseudomonas spp. other than fluorescent pseudomonads in mesophilic anaerobic digestion, the risk of spread of resistant bacteria to the environment should be considered when using digestate as liquid fertilizer.

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