Nitrogen dynamics and biological processes in soil amended with microalgae grown in abattoir digestate to recover nutrients.

The use of microalgae for nutrient recovery from wastewater and subsequent conversion of the harvested biomass into fertilizers offers a sustainable approach towards creating a circular economy. Nonetheless, the process of drying the harvested microalgae represents an additional cost, and its impact on soil nutrient cycling compared to wet algal biomass is not thoroughly understood. To investigate this, a 56-day soil incubation experiment was conducted to compare the effects of wet and dried Scenedesmus sp. microalgae on soil chemistry, microbial biomass, CO2 respiration, and bacterial community diversity. The experiment also included control treatments with glucose, glucose + ammonium nitrate, and no fertilizer addition. The Illumina Mi-Seq platform was used to profile the bacterial community and in-silico analysis was performed to assess the functional genes involved in N and C cycling processes. The maximum CO2 respiration and microbial biomass carbon (MBC) concentration of dried microalgae treatment were 17% and 38% higher than those of paste microalgae treatment, respectively. NH4+ and NO3- released slowly and through decomposition of microalgae by soil microorganisms as compared to synthetic fertilizer control. The results indicate that heterotrophic nitrification may contribute to nitrate production for both microalgae amendments, as evidenced by low amoA gene abundance and a decrease in ammonium with an increase in nitrate concentration. Additionally, dissimilatory nitrate reduction to ammonium (DNRA) may be contributing to ammonium production in the wet microalgae amendment, as indicated by an increase in nrfA gene and ammonium concentration. This is a significant finding because DNRA leads to N retention in agricultural soils instead of N loss via nitrification and denitrification. Thus, further processing the microalgae through drying or dewetting may not be favorable for fertilizer production as the wet microalgae appeared to promote DNRA and N retention.

Combining frass and fatty acid co-products derived from Black soldier fly larvae farming shows potential as a slow release fertiliser.

Use of black soldier fly larvae (BSFL) to process large volumes of organic waste is an emerging industry to produce protein. A co-product of this industry, the larval faeces (frass), has potential to be used as an organic fertiliser in a circular economy. However, BSFL frass has a high ammonium (N-NH4+) content which could result in nitrogen (N) loss following its application to land. One solution is to process the frass by combining it with solid fatty acids (FA) that have previously been used to manufacture slow-release inorganic fertilisers. We investigated the slow-releasing effect of N after combining BSFL frass with three FAs - lauric, myristic and stearic acid. Soil was amended with the three forms of FA processed (FA-P) frass, unprocessed frass or a control and incubated for 28 days. The impact of treatments on soil properties and soil bacterial communities were characterised during the incubation. Lower N-NH4+ concentrations occurred in soil treated with FA-P frass compared to unprocessed frass, and N-NH4+ release was slowest for lauric acid processed frass. Initially, all frass treatments caused a large shift in the soil bacterial community towards a dominance of fast-growing r-strategists that were correlated with increased organic carbon levels. FA-P frass appeared to enhance the immobilisation of N-NH4+ (from frass) by diverting it into microbial biomass. Unprocessed and stearic acid processed frass became enriched by slow-growing K-strategist bacteria at the latter stages of the incubation. Consequently, when frass was combined with FAs, FA chain length played an important role in regulating the composition of r-/K- strategists in soil and N and carbon cycling. Modifying frass with FAs could be developed into a slow release fertiliser leading to reduced soil N loss, improved fertiliser use efficiency, increased profitability and lower production costs.

Development of a controlled release fertilizer by incorporating lauric acid into microalgal biomass: Dynamics on soil biological processes for efficient utilisation of waste resources.

Utilisation of microalgae to extract nutrients from the effluent of anaerobic digestion of food waste is an emerging technology. A by-product of this process is the microalgal biomass which has potential to be used as an organic bio-fertilizer. However, microalgal biomass are rapidly mineralized when applied to soil which may result in N loss. One solution is to emulsify microalgal biomass with lauric acid (LA) to delay the release of mineral N. This study aimed to investigate whether combining LA with microalgae to develop a new fertilizer product with a controlled release function of mineral N when applied to soil, and any potential impacts the bacterial community structure and activity. The treatments were applied to soil emulsified with LA and were combined with either microalgae or urea at rates of 0%, 12.5%, 25% and 50% LA, untreated microalgae or urea and unamended control were incubated at 25 °C and 40% water holding capacity for 28 days. Quantification of soil chemistry (NH4+-N, NO3--N, pH and EC), microbial biomass carbon, CO2 production and bacterial diversity were characterised at 0, 1, 3, 7, 14 and 28 days. The NH4+-N and NO3--N concentration decreased with increasing rate of LA combined microalgae indicating that both N mineralization and nitrification were impacted. As a function of time, NH4+-N concentration increased up to 7 days for the microalgae at lower rates of LA, and then slowly decreased for 14 and 28 days, with an inverse relationship with soil NO3-N. Aligning with soil chemistry, an observed decrease in the predicted nitrification genes amoA·amoB and relative abundance of ammonia oxidizing bacteria (Nitrosomonadaceae) and nitrifying bacteria (Nitrospiraceae) with an increasing rate of LA with microalgae provides further support for possible inhibition of nitrification. The MBC and CO2 production was higher in the soil amended with increasing rates of LA combined microalgae and there was an increase in the relative abundance of fast-growing heterotrophs. Treating microalgae by emulsification with LA has the potential to control the release of N by increasing immobilization over nitrification and therefore it might be possible to engineer microalgae to match plant nutrient growth requirements whilst recovering waste from waste resources.

Co-application of a biosolids product and biochar to two coarse-textured pasture soils influenced microbial N cycling genes and potential for N leaching.

Co-application of biochar and biosolids to soil has potential to mitigate N leaching due to physical and chemical properties of biochar. Changes in N cycling pathways in soil induced by co-application of biological amendments could further mitigate N loss, but this is largely unexplored. The aim of this study was to determine whether co-application of a biochar and a modified biosolids product to three pasture soils differing in texture could alter the relative abundance of N cycling genes in soil sown with subterranean clover. The biosolids product contained lime and clay and increased subterranean clover shoot biomass in parallel with increases in soil pH and soil nitrate. Its co-application with biochar similarly increased plant growth and soil pH with a marked reduction in nitrate in two coarse textured soils but not in a clayey soil. While application of the biosolids product altered in silico predicted N cycling functional genes, there was no additional change when applied to soil in combination with biochar. This supports the conclusion that co-application of the biochar and biosolids product used here has potential to mitigate loss of N in coarse textured soils due to N adsoption by the biochar and independently of microbial N pathways.

Microalgae and Phototrophic Purple Bacteria for Nutrient Recovery From Agri-Industrial Effluents: Influences on Plant Growth, Rhizosphere Bacteria, and Putative Carbon- and Nitrogen-Cycling Genes.

Microalgae (MA) and purple phototrophic bacteria (PPB) have the ability to remove and recover nutrients from digestate (anaerobic digestion effluent) and pre-settled pig manure that can be Utilized as bio-fertilizer and organic fertilizer. The objective of this study was to compare the effectiveness of MA and PPB as organic fertilizers and soil conditioners in relation to plant growth and the soil biological processes involved in nitrogen (N) and carbon (C) cycling. To this end, a glasshouse experiment was conducted using MA and PPB as bio-fertilizers to grow a common pasture ryegrass (Lolium rigidum Gaudin) with two destructive harvests (45 and 60 days after emergence). To evaluate the rhizosphere bacterial community, we used barcoded PCR-amplified bacterial 16S rRNA genes for paired-end sequencing on the Illumina Mi-Seq. Additionally, we used phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt) analysis for the detection of putative functional genes associated with N and soil-C cycling. There was a significant increase in plant growth when the soil was amended with PPB, which almost performed as well as the chemical fertilizers. Analysis of the rhizosphere bacteria after the second harvest revealed a greater abundance of Firmicutes than in the first harvest. Members of this phylum have been identified as a biostimulant for plant growth. In contrast, the MA released nutrients more slowly and had a profound effect on N cycling by modulating N mineralization and N retention pathways. Thus, MA could be developed as a slow-release fertilizer with better N retention, which could improve crop performance and soil function, despite nutrient losses from leaching, runoff, and atmospheric emissions. These data indicate that biologically recovered nutrients from waste resources can be effective as a fertilizer, resulting in enhanced C- and N-cycling capacities in the rhizosphere.

Ammonia stress on a resilient mesophilic anaerobic inoculum: Methane production, microbial community, and putative metabolic pathways.

Short term inhibition tests, 16S rRNA tag sequencing and Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt), were employed to visualise the effects of increasing total ammoniacal nitrogen (TAN) concentration (3400-10166 ppm TAN) on microbial community structure and metabolic pathways for acetate degradation. The rate of methane production on acetate was significantly reduced by TAN concentrations above 6133 ppm; however, methane continued to be produced, even at 10166 ppm TAN (0.026 ±â€¯0.0003 gCOD.gVS-1inoculum.day-1). Hydrogenotrophic methanogenesis with syntrophic acetate oxidation (SAO) was identified as the dominant pathway for methane production. A shift towards SAO pathways at higher TAN concentrations and a decrease in the number of 'gene hits' for key genes in specific methanogenesis pathways was observed. Overall, the results highlighted potential for inhibition activity testing to be used together with PICRUSt, to estimate changes in microbial metabolism and to better understand microbial resilience in industrial AD facilities.

Amending Poultry Broiler Litter to Prevent the Development of Stable Fly, Stomoxys calcitrans (Diptera: Muscidae) and Other Nuisance Flies.

Spent poultry litter use as a fertilizer in horticulture supports stable fly Stomoxys calcitrans (L.) (Diptera: Muscidae) development. Stable fly continues to have an economic impact on livestock production and rural lifestyle in south-western Australia. The use of raw poultry manure is banned in 12 Shires surrounding Perth. The loss of market options for West Australian broiler growers has caused economic hardship. Hence, this study examined a range of chemical and biological amendments to spent poultry broiler litter in preventing stable fly and nuisance fly development. These included alkalizers (i.e., lime sand, quicklime, soda ash, and shell grit), acidifiers (aluminum sulfate, sodium bisulfate), gypsum, zeolite, spongolite, calcium cyanamide, and two fungal agents. The treated litters were placed under irrigation in horticulture with amendments added prior to them being exposed in the field as replicate 1-liter pads. In total, 19,559 stable flies developed from the spent litters exposed over five field experiments (88.7% of all flies recovered). House flies (Musca domestica L. (Diptera: Muscidae); 2,067 or 9.4%), false stable flies (Muscina stabulans Fallén (Diptera: Muscidae); 414 or 1.9%), and two sarcophagids (flesh fly) also developed from the litter. Borax completely prevented any fly development from the litter. Calcium cyanamide (1-2.5% v/v) and sodium bisulfate (10%) reduced stable fly numbers by as much as 99-100% when added to litter. Alkalizers, zeolite, spongolite, and entomopathogenic fungi had no significant impact on stable fly development. The addition of either calcium cyanamide or sodium bisulfate to raw litter can boost the fertilizer value of the litter while preventing stable fly development.

Ancient landscapes and the relationship with microbial nitrification.

Ammonia oxidizing archaea (AOA) and bacteria (AOB) drive nitrification and their population dynamics impact directly on the global nitrogen cycle. AOA predominate in the majority of soils but an increasing number of studies have found that nitrification is largely attributed to AOB. The reasons for this remain poorly understood. Here, amoA gene abundance was used to study the distribution of AOA and AOB in agricultural soils on different parent materials and in contrasting geologic landscapes across Australia (n = 135 sites). AOA and AOB abundances separated according to the geologic age of the parent rock with AOB higher in the more weathered, semi-arid soils of Western Australia. AOA dominated the younger, higher pH soils of Eastern Australia, independent of any effect of land management and fertilization. This differentiation reflects the age of the underlying parent material and has implications for our understanding of global patterns of nitrification and soil microbial diversity. Western Australian soils are derived from weathered archaean laterite and are acidic and copper deficient. Copper is a co-factor in the oxidation of ammonia by AOA but not AOB. Thus, copper deficiency could explain the unexpectedly low populations of AOA in Western Australian soils.

Microbial community dynamics in mesophilic anaerobic co-digestion of mixed waste.

This paper identifies key components of the microbial community involved in the mesophilic anaerobic co-digestion (AD) of mixed waste at Rayong Biogas Plant, Thailand. The AD process is separated into three stages: front end treatment (FET); feed holding tank and the main anaerobic digester. The study examines how the microbial community structure was affected by the different stages and found that seeding the waste at the beginning of the process (FET) resulted in community stability. Also, co-digestion of mixed waste supported different bacterial and methanogenic pathways. Typically, acetoclastic methanogenesis was the major pathway catalysed by Methanosaeta but hydrogenotrophs were also supported. Finally, the three-stage AD process means that hydrolysis and acidogenesis is initiated prior to entering the main digester which helps improve the bioconversion efficiency. This paper demonstrates that both resource availability (different waste streams) and environmental factors are key drivers of microbial community dynamics in mesophilic, anaerobic co-digestion.

Actinobacterial community dynamics in long term managed grasslands.

Palace Leas, a long-term experiment at Cockle Park Farm, Northumberland, UK was established in winter 1896-1897 since when the 13 plots have received regular and virtually unchanged mineral fertiliser and farm yard manure inputs. Fertilisers have had a profound impact on soil pH with the organically fertilised plots showing a significantly higher pH than those receiving mineral fertiliser where ammonium sulphate has led to soil acidification. Here, we investigate the impact of organic and mineral fertilisers on the actinobacterial community structure of these soils using terminal restriction fragment length polymorphism (T-RFLP) and 16S rRNA gene analysis. To differentiate fertiliser effects from seasonal variation, soils were sampled three times over one growing season between May and September 2004 and January 2005. Community profiles obtained using T-RFLP were analysed using multivariate statistics to investigate the relationship between community structure, seasonality and fertiliser management. Soil pH was shown to be the most significant edaphic factor influencing actinobacterial communities. Canonical correspondence analysis, used to investigate the relationship between the 16S rRNA gene community profiles and the environmental parameters, showed that actinobacterial communities also responded to soil water content with major changes evident over the summer months between May and September. Quantitative PCR of the actinobacterial and fungal 16S and 18S rRNA genes, respectively suggested that fungal rRNA gene copy numbers were negatively correlated (P = 0.0131) with increasing actinobacterial signals. A similar relationship (P = 0.000365) was also evident when fatty acid methyl esters indicative of actinobacterial biomass (10-methyloctadecanoic acid) were compared with the amounts of fungal octadecadienoic acid (18:2omega9,12). These results show clearly that soil pH is a major driver of change in actinobacterial communities and that genera such as Arthrobacter and Micrococcus are particularly abundant in soils receiving organic inputs whilst others such as Streptomyces, Acidimicrobium and Actinospica are more prevalent in acid soils. The importance of these findings in terms of fungal abundance and potential disease suppression are discussed.