Characterization of Nitrate-Reducing and Amino Acid–Using Bacteria Prominent in Nitrotoxin-Enriched Equine Cecal Populations

Horses can be at risk for nitropoisoning by consuming plants containing 3-nitro-1-propionic acid or 3-nitro-1-propanol and to a lesser extent by plants containing nitrate. Populations of equine cecal microbes enriched for enhanced rates of 3-nitro-1-propionic acid (NPA) or nitrate metabolism were cultured for NPA- or nitrate-metabolizing bacteria on basal enrichment medium or tryptose soy agar supplemented with either 5-mM NPA or nitrate and under H₂:CO₂ (20:80) as the energy source. After 72 hours, separated colonies picked from plates, or roll tubes were cultured in fresh broth medium for 72 hours and then identified by 16S rRNA gene sequencing. Isolates from the NPA-enriched populations were identified as Streptococcus lutetiensis (five strains), Escherichia coli (two strains), and Sporanaerobacter acetigens (one strain). Strains isolated from nitrate-enriched populations were identified as Escherichia coli (one strain) and Wolinella succinogenes (three strains). None of these strains degraded NPA. Enriched populations of equine cecal microbes, the isolated pure strains and the type strain of Denitrobacterium detoxificans, a competent NPA-metabolizing microbe, were examined using denaturing gradient gel electrophoresis (DGGE). The DGGE analysis indicated that none of the strains in the enriched population of equine cecal microbes were similar to D. detoxificans. However, we report for the first time the isolation of the anaerobic amino acid–using Sporanaerobacter acetigenes from the equine cecum.     

The routes to fruit: Governance of urban food trees in Canada

Interest in planting urban food trees (UFTs) in public spaces is growing in popularity as a form of urban greening and a potential food source. Currently there is minimal research on the governance and policy aspects of integrating food trees into cities. To fill this gap, we investigated the characteristics of UFT site governance and how it compares to current urban forest governance. Semi-structured interviews were conducted with relevant municipal officials in Calgary, Edmonton, Toronto and Victoria about their perspectives and involvement with UFT sites in their city. A scan of policy documents was completed to supplement the interviews. The interviews were analyzed using a deductive coding framework based on the Policy Arrangement Approach. We found that key actors at the UFT sites were local organizations and site champions, with minimal municipal engagement. Most site resources provided by municipalities were in-kind. There are also basic knowledge gaps about how to care for UFTs. Currently municipal by-laws prohibit the harvesting and removal of plant material, and are at odds with the purpose of UFT sites, with few municipalities fully integrating UFTs in policies. The primary discussion around UFTs centre concerns for public health and safety, management, and use of public space with limited discussions of benefits. This research demonstrates the value of co-governance models to support UFTs, while a shift in focus from risks to benefits could encourage additional resources and policy integration. Further, including UFTs into policy would also support foraging and food tree maintenance in public spaces, and more fully reflect the plurality of urban forest engagement.     

Citizens’ perception of and satisfaction with urban forests and green space: Results from selected Southeast European cities

In this paper the results of the first comprehensive study on perception of and satisfaction with urban forests and green space in seven Southeast European cities are presented. The aims of the paper are to analyse 1) citizen perceptions of the current state of urban forests and green space in their cities, 2) to what extent current urban forests and green space meet their needs and how this can be improved. A cross-sectional study was conducted by using a common face-to-face survey questionnaire. Respondents were selected based on census data though a stratified sampling procedure by taking into account age, gender and city district (n = 384 in each city). The results showed that citizens genuinely care for urban forests and green space in their cities, but are not satisfied with their current state. The respondents found issues related to misbehaviour of other users, the presence and quality of facilities, as well as the presence and quality of management or maintenance the most pressing. There were more statistically significant differences than similarities between cities. Socioeconomic variables explained perceptions only to some extent. Citizens were very supportive of educational campaigns about the importance of urban forests and green space as well as of better enforcement of the existing regulations though having more community wardens that were expected to tackle current unsatisfactory situations. Urban planning and urban forest and green space management in these cities are facing many problems characteristic for post-socialist countries. Study findings are expected to contribute to decision making in urban planning and natural resource management.     

Dense planting with less basal nitrogen fertilization might benefit rice cropping for high yield with less environmental impacts

Dense planting and less basal nitrogen (N) fertilization have been recommended to further increase rice (Oryza sativa L.) grain yield and N use efficiency (NUE), respectively. The objective of this study was to evaluate the integrative impacts of dense planting with reduced basal N application (DR) on rice yield, NUE and greenhouse gas (GHG) emissions. Field experiments with one conventional sparse planting (CK) and four treatments of dense planting (increased seedlings per hill) with less basal N application were conducted in northeast China from 2012 to 2013. In addition, a two-factor experiment was conducted to isolate the effect of planting density and basal N rate on CH₄ emission in 2013. Our results show that an increase in planting density by about 50% with a correspondingly reduction in basal N rate by about 30% (DR1 and DR2) enhanced NUE by 14.3–50.6% and rice grain yield by 0.5–7.4% over CK. Meanwhile, DR1 and DR2 reduced GWP by 6.4–12.6% and yield-scaled GWP by 7.0–17.0% over CK. According to the two-factor experiment, soil CH₄ production and oxidation and CH₄ emission were not affected by planting density. However, reduced basal N rate decreased CH₄ emission due to it significantly reduced soil CH₄ production with a smaller reduction in soil CH₄ oxidation. The above results indicate that moderate dense planting with less basal N application might be an environment friendly mode for rice cropping for high yield and NUE with less GHG emissions.     

Fungal and bacterial N2O production regulated by soil amendments of simple and complex substrates

Fungal N₂O production results from a respiratory denitrification that reduces NO₃⁻/NO₂⁻ in response to the oxidation of an electron donor, often organic C. Despite similar heterotrophic nature, fungal denitrifiers may differ from bacterial ones in exploiting diverse resources. We hypothesized that complex C compounds and substances could favor the growth of fungi over bacteria, and thereby leading to fungal dominance for soil N₂O emissions. Effects of substrate quality on fungal and bacterial N₂O production were, therefore, examined in a 44-d incubation after soils were amended with four different substrates, i.e., glucose, cellulose, winter pea, and switchgrass at 2 mg C g⁻¹ soil. During periodic measurements of soil N₂O fluxes at 80% soil water-filled pore space and with the supply of KNO₃, substrate treatments were further subjected to four antibiotic treatments, i.e., no antibiotics or soil addition of streptomycin, cycloheximide or both so that fungal and bacterial N₂O production could be separated. Up to d 8 when antibiotic inhibition on substrate-induced microbial activity and/or growth was still detectable, bacterial N₂O production was generally greater in glucose- than in cellulose-amended soils and also in winter pea- than in switchgrass-amended soils. In contrast, fungal N₂O production was more enhanced in soils amended with cellulose than with glucose. Therefore, fungal-to-bacterial contribution ratios were greater in complex than in simple C substrates. These ratios were positively correlated with fungal-to-bacterial activity ratios, i.e., CO₂ production ratios, suggesting that substrate-associated fungal or bacterial preferential activity and/or growth might be the cause. Considering substrate depletion over time and thereby becoming limited for microbial N₂O production, measurements of soil N₂O fluxes were also carried out with additional supply of glucose, irrespective of different substrate treatments. This measurement condition might lead to potentially high rates of fungal and bacterial N₂O production. As expected, bacterial N₂O production was greater with added glucose than with added cellulose on d 4 and d 8. However, this pattern was broken on d 28, with bacterial N₂O production lower with added glucose than with added cellulose. In contrast, plant residue impacts on soil N₂O fluxes were consistent over 44-d, with greater bacterial contribution, lower fungal contribution, and thus lower fungal-to-bacterial contribution ratios in winter pea- than in switchgrass-amended soils. Real-time PCR analysis also demonstrated that the ratios of 16S rDNA to ITS and the copy numbers of bacterial denitrifying genes were greater in winter pea- than in switchgrass-amended soils. Despite some inconsistency found on the impacts of cellulose versus glucose on fungal and bacterial leading roles for N₂O production, the results generally supported the working hypothesis that complex substrates promoted fungal dominance for soil N₂O emissions.     

Estimation irrigation water requirements with derived crop coefficients for upland and paddy crops in ChiaNan Irrigation Association,Taiwan

Field experiments were performed at the HsuehChia Experimental Station from 1993 to 2001 to calculate the reference and actual crop evapotranspiration, derived the crop coefficient, and collected requirements input data for the CROPWAT irrigation management model to estimate the irrigation water requirements of paddy and upland crops at the ChiaNan Irrigation Association, Taiwan. For corn, the estimated crop coefficients were 0.40, 0.78, 0.89 and 0.71 in the initial, crop development, mid-season and late-season stages, respectively. Meanwhile, the estimated crop coefficients for sorghum were 0.44, 0.71, 0.87 and 0.62 in the four stages, respectively. Finally, for soybean, the estimated crop coefficients were 0.45, 0.89, 0.92 and 0.58 in the four stages, respectively. With implementation of REF-ET model and FAO 56 Penman–Monteith method, the annual reference evapotranspiration was 1268 mm for ChiaNan Irrigation Association.In the paddy fields, the irrigation water requirements and deep percolation are 962 and 295 mm, respectively, for the first rice crop, and 1114 and 296 mm for the second rice crop. Regarding the upland crops, the irrigation water requirements for spring and autumn corn are 358 and 273 mm, respectively, compared to 332 and 366 mm for sorghum, and 350 and 264 mm for soybean. For the irrigated scheme with single and double rice cropping patterns in the ChiaNan Irrigation Association, the CROPWAT model simulated results indicate that the annual crop water demands are 507 and 1019 mm, respectively, and the monthly water requirements peaked in October at 126 mm and in January at 192 mm, respectively.     

基于生态理念的蚌埠黑虎山景区规划探析

城市边缘风景名胜区是城市大环境绿化的龙头,对改善城市生态环境具有重要意义。以蚌埠黑虎山景区规划为例,在概述黑虎山景区现状及发展优劣势的基础上,提出了基于生态理念的城市边缘风景名胜区的规划思想和方法,以期为城市边缘风景名胜区的建设和发展提供思路。     

Response of ammonia oxidizing microbes to the stresses of arsenic and copper in two acidic alfisols

Soil pollution by elevated heavy metals exhibits adverse effects on soil microorganisms. Ammonia oxidizing bacteria and ammonia oxidizing archaea perform ammonia oxidative processes in acidic soils. However, influence of heavy metal stress on soil ammonia oxidizers distribution and diversity is inadequately addressed. This study investigated the responses of ammonia oxidizing bacteria and archaea to heavy metals, Cu and As during short-term laboratory experiment. Two different acidic alfisols named as Rayka and Hangzhou spiked with different concentrations of As, Cu and As + Cu were incubated for 10 weeks. Significant reduction in copy numbers of archaeal-16S rRNA, bacterial-16S rRNA and functional amoA genes was observed along elevated heavy metal concentrations. Ammonia oxidizing archaea was found to be more abundant than ammonia oxidizing bacteria in all the heavy metal treatments. The potential nitrification rate significantly decreased with increasing As and Cu concentrations in the two soils examined. Denaturing gradient gel electrophoresis analysis revealed no apparent community shift for ammonia oxidizing archaea even at higher concentrations of As and Cu. Phylogenetic analysis of archaeal amoA gene from 4 clone libraries indicated that all the archaeal amoA sequences were placed within 3 distinct clusters from soil and sediment group 1.1b of Thaumarchaeota. Our results could be useful for the better understanding of the ecological effects of heavy metals on the abundance and diversity of soil ammonia oxidizers.