Mycovorax composti gen. nov., sp. nov., a member of the family Chitinophagaceae isolated from button mushroom compost.

Two Gram-stain-negative, aerobic, rod-shaped, orange-coloured bacterial strains, designated strain C216T and strain M2295, were isolated from mature mushroom compost from composting facilities in Victoria and South Australia, Australia, respectively. External structures such as flagella or pili were not observed on the cells under scanning electron microscopy. Optimal growth was found to occur at 45 °C, at pH 7.25 and in the absence of NaCl on Emerson's 350 YpSs medium. The genome sequence of strain C216T was 3 342 126 bp long with a G+C content of 40.5 mol%. Functional analysis of the genome of strain C216T revealed genes encoding chitinolytic and hemi-cellulolytic functions, with 166 predicted genes associated with carbohydrate metabolism (8.9% of the predicted genes). These functions are important for survival in the mushroom compost environment, which is rich in hemicelluloses. No antibiotic resistance genes were found in the genome sequence. The major fatty acids of strain C216T were iso-C15 : 0 (56.7%), iso-C17 : 0 3-OH (15.6%), C16 : 1 ω7c/iso-C15 : 0 2-OH (7.3%) and iso-C15 : 1 G (6.1%). The only respiratory quinone was MK-7. The major polar lipid of strain C216T was phosphatidylethanolamine, but three unidentified phospholipids, four unidentified aminophospholipids/aminolipids and one unidentified glycolipid were also detected. Phylogenetic analysis based on proteins encoded by the core genome (bac120, 120 conserved bacterial genes) showed that strain C216T forms a distinct lineage in the family Chitinophagaceae and that the closest identified relative is Niabella soli (69.69% ANI). These data demonstrate that strain C216T represents a novel genus and novel species within the family Chitinophagaceae, for which we propose the name Mycovorax composti. The type strain is C216T (=DSM 114558T=LMG 32998T).

Mixing postharvest fungicides and sanitizers results in unpredictable survival of microbes that affect cantaloupes.

Postharvest treatments with sanitizers and fungicides are applied to increase the quality, safety and shelf life of fresh produce including cantaloupes (also known as rockmelons). The primary role of sanitizers during cantaloupe washing is to prevent cross contamination of potentially pathogenic bacteria in washwater. Postharvest fungicide sprays or dips are employed to inhibit spoilage-causing fungi. While assessing the compatibility of these antimicrobials based on the measurement of active ingredients levels provides some indication of antimicrobial capacity, there is limited data on whether the interaction between these chemicals in wash water modifies their overall efficacy against relevant microorganisms. The aim of this research was to determine how chlorine- and peroxyacetic acid-based sanitizers interact with commercial guazatine- and imazalil-based fungicide formulations used on cantaloupes, and whether mixing these augments or suppresses anti-microbial activity against relevant human pathogens and spoilage fungi in wash water. The results were unpredictable: while most combinations were antimicrobial, the chlorine-based sanitizer when mixed with the guazatine-based fungicide had significantly reduced efficacy against pathogenic Salmonella spp. (~2.7 log) and the fungal spoilage organisms, Trichothecium roseum and Rhizopus stolonifera. Mixing the chlorine-based sanitizer with an imazalil-based fungicide produced a range of outcomes with antagonistic, indifferent and synergistic interactions observed for the fungal species tested. The peroxyacetic acid-based sanitizer led to indifferent interactions with the guazatine-based fungicide, while antagonism and synergy were observed when mixed with the imazalil-based fungicide. This study demonstrates that mixing postharvest agrichemicals used in the cantaloupe industry may increase the risk of microbial contamination and thereby potentially compromise food safety and quality.

Caffeine metabolism during cultivation of oyster mushroom (Pleurotus ostreatus) with spent coffee grounds.

In coffee-producing countries, waste products from coffee production are useful substrates for cultivation of Pleurotus ostreatus. This species is relatively easy to grow, coffee waste substrates are readily available and the mushroom fruiting bodies are a valuable source of nutrition and income. In developed countries, cultivation of P. ostreatus on spent coffee grounds (SCG) from coffee consumption is a novel way to recycle this urban waste product. Here, we studied the effect of SCG and caffeine on growth of a commercial strain of P. ostreatus in liquid and solid cultures, and on a commercial scale. The presence of caffeine inhibited mycelial growth on agar and in liquid culture in the laboratory. Increased levels of SCG in an SCG/sawdust substrate also delayed mycelial growth and delayed or prevented fruiting during commercial cultivation. Despite growth inhibition, partial degradation of caffeine to xanthine by P. ostreatus mycelium was observed in all SCG-containing substrate mixtures. Degradation of caffeine proceeded mainly via sequential N-demethylation to theophylline (1,3-dimethylxanthine) and 3-methylxanthine, although both paraxanthine and theobromine also accumulated in the substrate. Caffeine and its demethylated metabolites were also detected in fruiting bodies, but it was not clear whether caffeine metabolism occurred in the fruiting bodies themselves or whether caffeine metabolites were translocated there from the mycelium. Based on the caffeine concentrations measured in fruiting bodies after growth with SCG, it would be necessary to consume ~ 250 kg of fresh oyster mushrooms to obtain the amount of caffeine equivalent to one cup of espresso coffee, suggesting that the health impact of caffeine in these mushrooms is low. However, the ability of P. ostreatus to degrade caffeine indicates that this and other species in this genus may have potential applications in detoxification of coffee production wastes.

Bacterial population dynamics in recycled mushroom compost leachate.

Mushrooms are an important food crop throughout the world. The most important edible mushroom is the button mushroom (Agaricus bisporus), which comprises about 30% of the global mushroom market. This species is cultivated commercially on a selective compost that is produced predominantly from wheat straw/stable bedding and chicken manure, at a moisture content of around 70% (w/w) and temperatures of up to 80 °C. Large volumes of water are required to achieve this moisture content, and many producers therefore collect leachate from the composting windrows and bunkers (known in the industry as "goody water") and reuse it to wet the raw ingredients. This has the benefit of recycling and saving water and has the potential to enrich beneficial microorganisms that stimulate composting, but also the risk of enhancing pathogen populations that could reduce productivity. Here, we show by 16S rRNA gene sequencing that mushroom compost leachate contains a high diversity of unknown microbes, with most of the species found affiliated with the phyla Firmicutes and Proteobacteria. However, by far the most abundant species was the thermophile Thermus thermophilus, which made up approximately 50% of the bacterial population present. Although the leachate was routinely collected and stored in an aerated central storage tank, many of the bacterial species found in leachate were facultative anaerobes. However, there was no evidence for sulfide production, and no sulfate-reducing bacterial species were detected. Because T. thermophilus is important in the high temperature phase of composting, the use of recycled leachate as an inoculum for the raw materials is likely to be beneficial for the composting process.

Optimisation of fuel reduction burning regimes for carbon, water and vegetation outcomes.

Fire plays a critical role in biodiversity, carbon balance, soil erosion, and nutrient and hydrological cycles. While empirical evidence shows that fuel reduction burning can reduce the incidence, severity and extent of unplanned fires in Australia and elsewhere, the integration of environmental values into fire management operations is not well-defined and requires further research and development. In practice, the priority for fuel reduction burning is effective mitigation of risk to life and property. Environmental management objectives, including maintenance of high quality water, reduction of CO2 emissions and conservation of biodiversity can be constrained by this priority. We explore trade-offs between fuel reduction burning and environmental management objectives and propose a framework for optimising fuel reduction burning for environmental outcomes.

Combustion influences on natural abundance nitrogen isotope ratio in soil and plants following a wildfire in a sub-alpine ecosystem.

This before-and-after-impact study uses the natural abundance N isotope ratio (δ(15)N) to investigate the effects of a wildfire on sub-alpine ecosystem properties and processes. We measured the (15)N signatures of soil, charred organic material, ash and foliage in three sub-alpine plant communities (grassland, heathland and woodland) in south-eastern Australia. Surface bulk soil was temporarily enriched in (15)N immediately after wildfire compared to charred organic material and ash in all plant communities. We associated the enrichment of bulk soil with fractionation of N during combustion and volatilization of N, a process that also explains the sequential enrichment of (15)N of unburnt leaves > ash > charred organic material in relation to duration and intensity of heating. The rapid decline in (15)N of bulk soil to pre-fire values indicates that depleted ash, containing considerable amounts of total N, was readily incorporated into the soil. Foliar δ(15)N also increased with values peaking 1 year post-fire. Foliar enrichment was foremost coupled with the release of enriched NH4(+) into the soil owing to isotopic discrimination during volatilization of soluble N and combustion of organic material. The mode of post-fire regeneration influenced foliar (15)N enrichment in two species indicating use of different sources of N following fire. The use of natural abundance of (15)N in soil, ash and foliage as a means of tracing transformation of N during wildfire has established the importance of combustion products as an important, albeit temporary source of inorganic N for plants regenerating after wildfire.

Panicum decompositum, an Australian Native Grass, Has Strong Potential as a Novel Grain in the Modern Food Market.

Native Millet (Panicum decompositum) is a native grass species that was used as a staple food by many Australian Aboriginal communities. In this study, the potential for using Native Millet (NM) as a novel flour in the modern food market was investigated. Intact grain and white and wholemeal flours from two populations of NM were compared to bread wheat cv. Spitfire (SW) using a range of physical and chemical tests. The baking properties of NM flour were assessed using basic flatbreads made with 25:75 and 50:50 (NM:SW) mixes of wholemeal flour with 100% SW wholemeal flour used as the control. The grain size of NM was found to be smaller than SW. Milling yield, defined as the proportion of flour obtained from a whole seed, for NM was 4-10% lower than SW under the same moisture conditions used for tempering (drying) wheat. The properties of wholemeal flour indicated that NM flour has lower viscosity and low flour pasting ability compared to SW. This is likely due to the low starch content and high fibre content of NM seed. Wholemeal flour derived from NM had a protein content of 13.6% compared to 12.1% for SW. Based on a sensory analysis using an untrained panel, the distinct colour and texture may negatively affect the acceptance of NM flour by the consumer, but taste and aroma was not found to differ among samples. There were strong indications that the novelty of NM flour may help outweigh any limitations to consumer acceptance, making it a valuable product in future food markets.

Dynamics of microbial community and enzyme activities during preparation of Agaricus bisporus compost substrate.

Button mushrooms (Agaricus bisporus) are grown commercially on a specialized substrate that is usually prepared from wheat straw and poultry manure in a microbially-mediated composting process. The quality and yield of the mushroom crop depends critically on the quality of this composted substrate, but details of the microbial community responsible for compost production have only emerged recently. Here we report a detailed study of microbial succession during mushroom compost production (wetting, thermophilic, pasteurization/conditioning, spawn run). The wetting and thermophilic phases were characterized by a rapid succession of bacterial and fungal communities, with maximum diversity at the high heat stage. Pasteurization/conditioning selected for a more stable community dominated by the thermophilic actinomycete Mycothermus thermophilus and a range of bacterial taxa including Pseudoxanthomonas taiwanensis and other Proteobacteria. These taxa decreased during spawn run and may be acting as a direct source of nutrition for the proliferating Agaricus mycelium, which has previously been shown to use microbial biomass in the compost for growth. Comparison of bacterial communities at five geographically separated composting yards in south-eastern Australia revealed similarities in microbial succession during composting, although the dominant bacterial taxa varied among sites. This suggests that specific microbial taxa or combinations of taxa may provide useful biomarkers of compost quality and may be applied as predictive markers of mushroom crop yield and quality.

Temperature-dependent release of volatile organic compounds of eucalypts by direct analysis in real time (DART) mass spectrometry.

A method is described for the rapid identification of biogenic, volatile organic compounds (VOCs) emitted by plants, including the analysis of the temperature dependence of those emissions. Direct analysis in real time (DART) enabled ionization of VOCs from stem and leaf of several eucalyptus species including E. cinerea, E. citriodora, E. nicholii and E. sideroxylon. Plant tissues were placed directly in the gap between the DART ionization source skimmer and the capillary inlet of the time-of-flight (TOF) mass spectrometer. Temperature-dependent emission of VOCs was achieved by adjusting the temperature of the helium gas into the DART ionization source at 50, 100, 200 and 300 degrees C, which enabled direct evaporation of compounds, up to the onset of pyrolysis of plant fibres (i.e. cellulose and lignin). Accurate mass measurements facilitated by TOF mass spectrometry provided elemental compositions for the VOCs. A wide range of compounds was detected from simple organic compounds (i.e. methanol and acetone) to a series of monoterpenes (i.e. pinene, camphene, cymene, eucalyptol) common to many plant species, as well as several less abundant sesquiterpenes and flavonoids (i.e. naringenin, spathulenol, eucalyptin) with antioxidant and antimicrobial properties. The leaf and stem tissues for all four eucalypt species showed similar compounds. The relative abundances of methanol and ethanol were greater in stem wood than in leaf tissue suggesting that DART could be used to investigate the tissue-specific transport and emissions of VOCs.

Nitrogen uptake by Eucalyptus regnans and Acacia spp. - preferences, resource overlap and energetic costs.

In southeastern Australia, the overstory species Eucalyptus regnans F. Muell. commonly grows with either of the two leguminous understory trees, Acacia melanoxylon (R. Br. Ex Ait. f.) or Acacia dealbata (Link.). Our objective was to elucidate interactions between the dominant eucalypt and its companion acacias for nitrogen (N) sources. Use of stable N isotopes as tracers revealed that ammonium was the preferred soil N source for all species, nevertheless, total N uptake varied greatly among species. Studies with double-labeled ((13)C/(15)N) glutamine indicated the uptake of this form of organic N in small amounts by both E. regnans and the Acacia spp. These and other data imply that, in contrast to boreal forests, organic N is not a significant component of N nutrition in mountain ash forests. Field and laboratory studies provided evidence that N(2)-fixation capacity of acacias varies with stand development, with N-fixing species playing an important role in N nutrition during the early but not the mature stages of forest growth. An index of N-uptake efficiency - the amount of oxygen consumed per unit N taken up - was compared across four N sources and three species. Nitrate uptake was the least efficient form of N acquisition, especially compared with ammonium uptake which was up to 30-fold less costly. Efficiency of glutamine uptake was intermediate between that of ammonium and nitrate. Differences in uptake efficiency among N forms were most pronounced for the Acacia spp. and least for E. regnans. We conclude that an overlap in requirements among sympatric Acacia spp. and E. regnans for specific soil N sources can be bypassed because of changes in biochemical strategies of Acacia spp. triggered by increasing soil N concentrations during stand development. Further studies might elucidate whether this is a common feature of complex forest ecosystems, or a specialty of the interaction between eucalypts and acacias.

N2-fixing black locust intercropping improves ecosystem nutrition at the vulnerable semi-arid Loess Plateau region, China.

The Loess Plateau in northwestern China constitutes one of the most vulnerable semi-arid regions in the world due to long-term decline in forest cover, soil nutrient depletion by agricultural use, and attendant soil erosion. Here, we characterize the significance of N2-fixing Robinia pseudoacacia L. and non-N2-fixing Juglans regia L. for improving nutrient availability and water retention in soil by comparing a range of biological and physicochemical features in monoculture and mixed plantations of both species. We found that N2-fixing Robinia facilitates the nitrogen and phosphorus composition of non-N2-fixing Juglans in the mixed stand as a consequence of improved soil nutrient availability, evident as higher levels of nitrogen and labile carbon compared to mono-specific stands. This demonstrates that intercropping N2-fixing Robinia with non-N2-fixing woody plants can greatly improve soil carbon and nitrogen bioavailability as well as whole-plant nutrition and can potentially mediate water retention with additional sequestration of soil organic carbon in the range of 1 t C ha-1 year-1. Thus, intercropping N2-fixing woody species (e.g. Robinia pseudoacacia or Hippophae rhamnoides L.) with locally important non-N2-fixing tree and shrub species should be considered in afforestation strategies for landscape restoration.

Can a growth model be used to describe forest carbon and water balance after fuel reduction burning in temperate forests?

Empirical evidence from Australia shows that fuel reduction burning significantly reduces the incidence and extent of unplanned fires. However, the integration of environmental values into fire management operations is not yet well-defined and requires further research and development. WAVES, a plant growth model that incorporates Soil-Vegetation-Atmosphere Transfer, was used to simulate the hydrological and ecological effects of three fuel management scenarios on a forest ecosystem. WAVES was applied using inputs from a set of forest plots for one year after three potential scenarios: (1) all litter removed, (2) all litter and 50% of the understorey removed, (3) all litter and understorey removed. Modelled outputs were compared with sites modelled with no-fuel reduction treatment (Unburnt). The key change between unburnt and fuel reduced forests was a significant increase in soil moisture after fire. Predictions of the recovery of aboveground carbon as plant biomass were driven by model structure and thus variability in available light and soil moisture at a local scale. Similarly, effects of fuel reduction burning on water processes were mainly due to changes in vegetation interception capacity (i.e. regrowth) and soil evaporation. Predicted effects of fuel reduction burning on total evapotranspiration (ET) - the major component of water balance - were marginal and not significant, even though a considerable proportion of ET had effectively been transferred from understorey to overstorey. In common with many plant growth models, outputs from WAVES are dictated by the assumption that overstorey trees continue to grow irrespective of their age or stage of maturity. Large areas of eucalypt forests and woodlands in SE Australia are well beyond their aggrading phase and are instead over-mature. The ability of these forests to rapidly respond to greater availability of water remains uncertain.