Koki Maeda

Source of nitrous oxide emissions during the cow manure composting process as revealed by isotopomer analysis of and amoA abundance in betaproteobacterial ammonia-oxidizing bacteria.

ABSTRACT A molecular analysis of betaproteobacterial ammonia oxidizers and a N2O isotopomer analysis were conducted to study the sources of N2O emissions during the cow manure composting process. Much NO2−-N and NO3−-N and the Nitrosomonas europaea-like amoA gene were detected at the surface, especially at the top of the composting pile, suggesting that these ammonia-oxidizing bacteria (AOB) significantly contribute to the nitrification which occurs at the surface layer of compost piles. However, the 15N site preference within the asymmetric N2O molecule (SP = δ15Nα − δ15Nβ, where 15Nα and 15Nβ represent the 15N/14N ratios at the center and end sites of the nitrogen atoms, respectively) indicated that the source of N2O emissions just after the compost was turned originated mainly from the denitrification process. Based on these results, the reduction of accumulated NO2−-N or NO3−-N after turning was identified as the main source of N2O emissions. The site preference and bulk δ15N results also indicate that the rate of N2O reduction was relatively low, and an increased value for the site preference indicates that the nitrification which occurred mainly in the surface layer of the pile partially contributed to N2O emissions between the turnings.

Characterization and spatial distribution of bacterial communities within passively aerated cattle manure composting piles

The bacterial communities in the core, bottom, top, middle-surface, and lower-surface full-scale passively aerated cattle manure compost was investigated using DGGE of PCR-amplified 16S rRNA sequences. Some Bacillus species and strictly anaerobic thermophilic Clostridium species were dominant only in the core and bottom zones. In contrast, bands belonging to mesophilic bacteria such as Bacteroidetes, Clostoridia,alpha and gamma-proteobacteria were detected in surface zones, even in the initial thermophilic stage of the process. Our results clearly show the spatial distribution of the microbial community within full-scale composting piles, which indicates N or C conversion by zone-specific bacterial communities were occurring in each zone of the pile.

Microbiology of nitrogen cycle in animal manure compost.

Composting is the major technology in the treatment of animal manure and is a source of nitrous oxide, a greenhouse gas. Although the microbiological processes of both nitrification and denitrification are involved in composting, the key players in these pathways have not been well identified. Recent molecular microbiological methodologies have revealed the presence of dominant Bacillus species in the degradation of organic material or betaproteobacterial ammonia‐oxidizing bacteria on nitrification on the surface, and have also revealed the mechanism of nitrous oxide emission in this complicated process to some extent. Some bacteria, archaea or fungi still would be considered potential key players, and the contribution of some pathways, such as nitrifier denitrification or heterotrophic nitrification, might be involved in composting. This review article discusses these potential microbial players in nitrification–denitrification within the composting pile and highlights the relevant unknowns through recent activities that focus on the nitrogen cycle within the animal manure composting process.

N2O production, a widespread trait in fungi.

N2O is a powerful greenhouse gas contributing both to global warming and ozone depletion. While fungi have been identified as a putative source of N2O, little is known about their production of this greenhouse gas. Here we investigated the N2O-producing ability of a collection of 207 fungal isolates. Seventy strains producing N2O in pure culture were identified. They were mostly species from the order Hypocreales order—particularly Fusarium oxysporum and Trichoderma spp.—and to a lesser extent species from the orders Eurotiales, Sordariales, and Chaetosphaeriales. The N2O 15N site preference (SP) values of the fungal strains ranged from 15.8‰ to 36.7‰, and we observed a significant taxa effect, with Penicillium strains displaying lower SP values than the other fungal genera. Inoculation of 15 N2O-producing strains into pre-sterilized arable, forest and grassland soils confirmed the ability of the strains to produce N2O in soil with a significant strain-by-soil effect. The copper-containing nitrite reductase gene (nirK) was amplified from 45 N2O-producing strains, and its genetic variability showed a strong congruence with the ITS phylogeny, indicating vertical inheritance of this trait. Taken together, this comprehensive set of findings should enhance our knowledge of fungi as a source of N2O in the environment.

Key odor components responsible for the impact on olfactory sense during swine feces composting

This study aimed to identify the major odor contributing components produced during swine feces composting which have an impact on the olfactory senses. A total of 64 gas samples collected at different stages of composting were analyzed by both a gas chromatograph and human panel test using the triangle odor bag method. Multiple regression analysis of representative odor substances present in the outlet gas was carried out employing the odor index (OI) as the dependent variable and the odor unit as the independent variable. The recorded changes in OI indicated that turning was an important event during odor evolution, and that the odor emission during the thermophilic phase should be the main target for odor abatement. The model incorporating ammonia, methyl mercaptan and dimethyl sulfide as independent variables confirmed the value of the OI (R(2)=0.70). These compounds were identified to be the key odor components significantly determining the OI.

The impact of using mature compost on nitrous oxide emission and the denitrifier community in the cattle manure composting process.

The diversity and dynamics of the denitrifying genes (nirS, nirK, and nosZ) encoding nitrite reductase and nitrous oxide (N2O) reductase in the dairy cattle manure composting process were investigated. A mixture of dried grass with a cattle manure compost pile and a mature compost-added pile were used, and denaturing gradient gel electrophoresis was used for denitrifier community analysis. The diversity of nirK and nosZ genes significantly changed in the initial stage of composting. These variations might have been induced by the high temperature. The diversity of nirK was constant after the initial variation. On the other hand, the diversity of nosZ changed in the latter half of the process, a change which might have been induced by the accumulation of nitrate and nitrite. The nirS gene fragments could not be detected. The use of mature compost that contains nitrate and nitrite promoted the N2O emission and significantly affected the variation of nosZ diversity in the initial stage of composting, but did not affect the variation of nirK diversity. Many Pseudomonas-like nirK and nosZ gene fragments were detected in the stage in which N2O was actively emitted.

Effect of covering composting piles with mature compost on ammonia emission and microbial community structure of composting process.

To control ammonia (NH(3)) volatilization from the dairy cattle (Bos taurus) manure composting process, a compost pile was covered with mature compost and the gas emissions evaluated using the dynamic chamber system. The peak of NH(3) volatilization observed immediately after piling up of the compost was reduced from 196 to 62 mg/m(3) by covering the compost pile with mature compost. The accumulation of NH(4)-N to the covered mature compost was also observed. Covering and mixing the compost with mature compost had no effect on the microbial community structure. However, over time the microbial community structure changed because of a decrease in easily degradable organic compounds in the compost piles. The availability of volatile fatty acids (VFA) was considered to be important for microbial community structure in the compost. After the VFA had disappeared, the NO(3)-N concentration increased and the cellulose degrading bacteria such as Cytophaga increased in number.

Mitigation of greenhouse gas emission from the cattle manure composting process by use of a bulking agent

The greenhouse gas (GHG) [methane (CH4) and nitrous oxide (N2O)] mitigation effects of mixing dried grass into passively-aerated manure during the composting process (which accounts for 68.7% of Japanese dairy manure management) were assessed. Gaseous emissions [CH4, N2O, carbon dioxide (CO2) and ammonia (NH3)] from about 4 t of fresh dairy manure with or without 400 kg of dried grass mixed in were measured by the dynamic chamber method. The addition of dried grass contributed to a decrease in GHG emissions from 20.8 ± 1.3 g kg−1 volatile solids (VS) to 5.4 ± 1.4 g kg−1 VS (74.3% mitigation) for CH4 and from 7.4 ± 2.6 g N2O-N kg−1 Ninitial to 2.7 ± 0.4 g N2O-N kg−1 Ninitial (62.8% mitigation) for N2O. By applying this strategy, the expected reduction of GHG emission would be 70,466 t CH4 yr−1 and 1379 t N2O-N yr−1 (1907 Gg CO2 eq. yr−1 in total) in the Japanese dairy sector. On the other hand, it was showed that CO2 and NH3 emissions increase [from 424.4 ± 214.9 g CO2 kg−1 VS to 603.8 ± 99.6 g CO2 kg−1 VS for CO2 and from 16.9 ± 7.1 g ammonium-nitrogen (NH3-N) kg−1 Ninitial to 38.3 ± 3.5 g NH3-N kg−1 Ninitial for NH3] by this method. Moreover, the mechanism of this significant N2O mitigation effect cannot be explained, and a better understanding of this effect could further improve the GHG mitigation strategy.

Denitrifiers in the surface zone are primarily responsible for the nitrous oxide emission of dairy manure compost

During the dairy manure composting process, significant nitrous oxide (N2O) emissions occur just after the pile turnings. To understand the characteristics of this N2O emission, samples were taken from the compost surface and core independently, and the N2O production was monitored in laboratory incubation experiments. Equal amounts of surface and core samples were mixed to simulate the turning, and the (15)N isotope ratios within the molecules of produced N2O were analyzed by isotopomer analysis. The results showed that the surface samples emitted significant levels of N2O, and these emissions were correlated with NOx(-)-N accumulation. Moreover, the surface samples and surface-core mixed samples incubated at 30°C produced N2O with a low site preference (SP) value (-0.9 to 7.0‰) that was close to bacteria denitrification (0‰), indicating that denitrifiers in the surface samples are responsible for this N2O production. On the other hand, N2O produced by NO2(-)-amended core samples and surface samples incubated at 60°C showed unrecognized isotopic signatures (SP=11.4-20.3‰). From these results, it was revealed that the N2O production occurring just after the turnings was mainly derived from bacterial denitrification (including nitrifier denitrification) of NOx(-)-N under mesophilic conditions, and surface denitrifying bacteria appeared to be the main contributor to this process.

Relative Contribution of nirK- and nirS- Bacterial Denitrifiers as Well as Fungal Denitrifiers to Nitrous Oxide Production from Dairy Manure Compost

The relative contribution of fungi, bacteria, and nirS and nirK denirifiers to nitrous oxide (N2O) emission with unknown isotopic signature from dairy manure compost was examined by selective inhibition techniques. Chloramphenicol (CHP), cycloheximide (CYH), and diethyl dithiocarbamate (DDTC) were used to suppress the activity of bacteria, fungi, and nirK-possessing denitrifiers, respectively. Produced N2O were surveyed to isotopocule analysis, and its 15N site preference (SP) and δ18O values were compared. Bacteria, fungi, nirS, and nirK gene abundances were compared by qPCR. The results showed that N2O production was strongly inhibited by CHP addition in surface pile samples (82.2%) as well as in nitrite-amended core samples (98.4%), while CYH addition did not inhibit the N2O production. N2O with unknown isotopic signature (SP = 15.3-16.2‰), accompanied by δ18O (19.0-26.8‰) values which were close to bacterial denitrification, was also suppressed by CHP and DDTC addition (95.3%) indicating that nirK denitrifiers were responsible for this N2O production despite being less abundant than nirS denitrifiers. Altogether, our results suggest that bacteria are important for N2O production with different SP values both from compost surface and pile core. However, further work is required to decipher whether N2O with unknown isotopic signature is mostly due to nirK denitrifiers that are taxonomically different from the SP-characterized strains and therefore have different SP values rather than also being interwoven with the contribution of the NO-detoxifying pathway and/or of co-denitrification.