Fast screening of total nutrient contents in strawberry leaves and spent growing media using NIRS.

Introduction: In closed-loop soilless cultivation, the main nutrient sinks are nutrients retained either by the crop or in spent growing media. Measurement of nutrients in spent growing media and in the aboveground vegetative plant biomass at crop termination can be a tool for assessing and optimizing nutrient efficiency. The first aim of this study was to test the potential of near-infrared reflectance spectroscopy (NIRS) to forecast the various nutrient contents in strawberry leaves, which would then allow for assessment of crop nutrient status and total nutrient uptake by strawberry plants. The second aim was to test NIRS as a high throughput technique for assessing the N, K, Ca, Mg and organic matter (OM) content and the pH, EC and C:N and C:P ratios for a dataset of composts, plant fibers and spent growing media. The NIRS prediction model for fast screening of the total nutrient contents in spent growing media was compared with a single extraction method. Methods: A database with 369 dried and ground strawberry leaf samples with known contents of N, P, K, Ca, and Mg were scanned using NIRS. The database covered a range of leaf contents of 6-35 g N/kg dry matter (DM), 0.7-6.3 g P/kg DM and 2-29 g K/kg DM. A dataset of 458 samples of different types of materials used in growing media was validated with a dataset of 109 samples. Results: Validation for the strawberry leaves indicated potential for this application, with R2 values of 0.90 or higher for N, K and Ca, and R2 values higher than 0.85 for P and Mg. Validation for the dataset of composts, plant fibers and spent growing media also indicated the potential for this application, with R2 values of 0.90 or higher for organic matter, and with R2 values of 0.85 or higher for total Ca, pH and C:N. A first test indicated potential for the calibration based on fresh samples of compost, plant fiber as well as spent growing media or dried (not ground) samples. Discussion: Use of NIRS on fresh samples would eliminate the need for drying and grinding the samples and would reduce screening time. The ammonium acetate extraction is a reliable alternative to NIRS for fast screening of the total P, K, Ca, and Mg contents in composts, plant fibers and spent growing media.

Oxygen uptake rate versus CO2 based respiration rate for assessment of the biological stability of peat, plant fibers and woody materials with high C:N ratio versus composts.

The biological stability of organic materials predicts their performance when used as either a soil improver or as an ingredient in growing media. CO2 release in a static measurement and O2 consumption rate (OUR) were compared for seven groups of growing media components. The ratio between CO2 release and OUR was matrix-specific. This ratio was highest for plant fibers high in C:N and with a high risk of N immobilization, intermediate for wood fiber and woody composts, and lowest for peat and other compost types. The effect of variable test conditions in the OUR setup was assessed for plant fibers, where addition of mineral N and/or nitrification inhibitor had no effect on the OUR measurements. Testing at 30 °C instead of 20 °C resulted in higher OUR values as expected, but did not change the effect of mineral N dose. A strong increase in the CO2 flux was measured when plant fibers were mixed with mineral fertilizer; in contrast, addition of mineral N or fertilizer before or during the OUR test had no effect. The present experimental setup did not allow for differentiation between a higher CO2 release as a result of increased microbial respiration after adding mineral N versus an underestimation of stability due to N limitation in the dynamic OUR setup. Results indicate that type of material, C:N ratio and risk of N immobilization all appear to affect the OUR results. The OUR criteria may therefore require clear differentiation based on the different materials used in horticultural substrates.

Reuse of coir, peat, and wood fiber in strawberry production.

Introduction: Production of strawberries in greenhouses and polytunnels is gaining popularity worldwide. This study investigated the effect of reuse of coir and peat, two substrates commonly adapted to soilless strawberry production, as well as stand-alone wood fiber from Norway spruce, a promising substrate candidate. Methods: The experiment was performed in a polytunnel at NIBIO Apelsvoll, Norway, and evaluated both virgin substrates, as well as spent materials that were used in one or two years. Yield, berry quality and plant architecture of the strawberry cultivar 'Malling Centenary' were registered. In addition, chemical and physical properties of virgin and reused substrates were investigated. Results: While plants grown in peat and wood fiber had highest yield in the first year of production, the berry yield was slightly reduced when these substrates were utilized for the second and third time. However, yield was comparable to the yield level attained in new and reused coir. Interestingly, berries grown in wood fiber had a tendency to a higher sugar accumulation. This substrate also produced the highest plants. Stand-alone wood fiber was the substrate with the highest accumulation of nitrogen during the three consecutive production cycles. All three investigated materials revealed a trend for decreased potassium accumulation. Wood fiber is characterized by the highest percentage of cellulose, however after three years of production the cellulose content was reducedto the same levels as for coir and peat. Discussion: Implementation of wood fiber as a growing medium, as well as general practice of substrate reuse can be therefore an achievable strategy for more sustainable berry production.

Elucidating the microbiome of the sustainable peat replacers composts and nature management residues.

Sustainable peat alternatives, such as composts and management residues, are considered to have beneficial microbiological characteristics compared to peat-based substrates. Studies comparing microbiological characteristics of these three types of biomass are, however, lacking. This study examined if and how microbiological characteristics of subtypes of composts and management residues differ from peat-based substrates, and how feedstock and (bio)chemical characteristics drive these characteristics. In addition, microbiome characteristics were evaluated that may contribute to plant growth and health. These characteristics include: genera associated with known beneficial or harmful microorganisms, microbial diversity, functional diversity/activity, microbial biomass, fungal to bacterial ratio and inoculation efficiency with the biocontrol fungus Trichoderma harzianum. Bacterial and fungal communities were studied using 16S rRNA and ITS2 gene metabarcoding, community-level physiological profiling (Biolog EcoPlates) and PLFA analysis. Inoculation with T. harzianum was assessed using qPCR. Samples of feedstock-based subtypes of composts and peat-based substrates showed similar microbial community compositions, while subtypes based on management residues were more variable in their microbial community composition. For management residues, a classification based on pH and hemicellulose content may be relevant for bacterial and fungal communities, respectively. Green composts, vegetable, fruit and garden composts and woody composts show the most potential to enhance plant growth or to suppress pathogens for non-acidophilic plants, while grass clippings, chopped heath and woody fractions of compost show the most potential for blends for calcifuge plants. Fungal biomass was a suitable predictor for inoculation efficiency of composts and management residues.

Biochar-Enhanced Resistance to Botrytis cinerea in Strawberry Fruits (But Not Leaves) Is Associated With Changes in the Rhizosphere Microbiome.

Biochar has been reported to play a positive role in disease suppression against airborne pathogens in plants. The mechanisms behind this positive trait are not well-understood. In this study, we hypothesized that the attraction of plant growth-promoting rhizobacteria (PGPR) or fungi (PGPF) underlies the mechanism of biochar in plant protection. The attraction of PGPR and PGPF may either activate the innate immune system of plants or help the plants with nutrient uptake. We studied the effect of biochar in peat substrate (PS) on the susceptibility of strawberry, both on leaves and fruits, against the airborne fungal pathogen Botrytis cinerea. Biochar had a positive impact on the resistance of strawberry fruits but not the plant leaves. On leaves, the infection was more severe compared with plants without biochar in the PS. The different effects on fruits and plant leaves may indicate a trade-off between plant parts. Future studies should focus on monitoring gene expression and metabolites of strawberry fruits to investigate this potential trade-off effect. A change in the microbial community in the rhizosphere was also observed, with increased fungal diversity and higher abundances of amplicon sequence variants classified into Granulicella, Mucilaginibacter, and Byssochlamys surrounding the plant root, where the latter two were reported as biocontrol agents. The change in the microbial community was not correlated with a change in nutrient uptake by the plant in either the leaves or the fruits. A decrease in the defense gene expression in the leaves was observed. In conclusion, the decreased infection of B. cinerea in strawberry fruits mediated by the addition of biochar in the PS is most likely regulated by the changes in the microbial community.

Understanding the Shift in the Microbiome of Composts That Are Optimized for a Better Fit-for-Purpose in Growing Media.

Three characteristics are considered key for optimal use of composts in growing media: maturity, pH and organic matter content. Maturation is a critical step in the processing of composts contributing to compost quality. Blending of composts with chopped heath biomass, sieving out the larger fraction of composts and acidification of composts by adding elemental sulfur may be used either to increase organic matter content or to reduce pH for a better fit in growing media. While several studies have shown the effectiveness of these treatments to improve the use of composts in growing media, the effect of these treatments on the compost microbiome has merely been assessed before. In the present study, five immature composts were allowed to mature, and were subsequently acidified, blended or sieved. Bacterial and fungal communities of the composts were characterized and quantified using 16S rRNA and ITS2 gene metabarcoding and phospholipid fatty acid analysis. Metabolic biodiversity and activity were analyzed using Biolog EcoPlates. Compost batch was shown to be more important than maturation or optimization treatments to determine the compost microbiome. Compost maturation increased microbial diversity and favored beneficial microorganisms, which may be positive for the use of composts in growing media. Blending of composts increased microbial diversity, metabolic diversity, and metabolic activity, which may have a positive effect in growing media. Blending may be used to modify the microbiome to a certain degree in order to optimize microbiological characteristics. Acidification caused a decrease in bacterial diversity and microbial activity, which may be negative for the use in growing media, although the changes are limited. Sieving had limited effect on the microbiome of composts. Because of the limited effect on the microbiome, sieving of composts may be used flexible to improve (bio)chemical characteristics. This is the first study to assess the effects of maturation and optimization treatments to either increase organic matter content or lower pH in composts on the compost microbiome.

Chemically versus thermally processed brown shrimp shells or Chinese mitten crab as a source of chitin, nutrients or salts and as microbial stimulant in soilless strawberry cultivation.

Brown shrimp (Crangon crangon) shells and Chinese mitten crab (Eriocheir sinensis) were chemically demineralized and deproteinized (denoted as M1 to M4 for the shrimp shells and M5 to M7 for the Chinese mitten crab), and shrimp shells were torrefied at 200 to 300 °C (denoted as R200, R255, R300), and were compared with a commercially available chitin source (denoted as reference chitin). Based on their chemical characteristics, a selection of chitin sources was tested for their N mineralization capacity. The N release was high for the chemically treated shrimp shells and Chinese mitten crab, but not for the torrefied shrimp shells with or without acid treatment, indicating that treatment at 200 °C or higher resulted in low N availability. Interaction with nutrients was tested in a leaching experiment with limed peat for three thermally and two chemically processed shrimp shells and the reference chitin source. The K concentrations in the leachate for the chemically treated shrimp shells and the reference chitin were lower than for limed peat during fertigation. Irreversible K retention was observed for one source of chemically treated shrimp shells, and the reference chitin. The thermally treated shrimp shells had a significantly higher net release of P, Na and Cl than the treatment without chitin source. Three shrimp shell based materials (M4, R200 and R300) and the reference chitin were tested in a greenhouse trial with strawberry at a dose of 2 g/L limed peat. A very positive and significant effect on Botrytis cinerea disease suppression in the leaves was found for the reference chitin, M4 and R200 compared to the unamended control. The disease suppression of the 3 chitin sources was linked with an increase of the microbial biomass in the limed peat with 24% to 28% due to chitin decomposition and a 9-44% higher N uptake in the plants.

Acidification of composts versus woody management residues: Optimizing biological and chemical characteristics for a better fit in growing media.

Previous research has demonstrated that composts (COM) and woody residues from nature management (MR) are potential peat replacers for growing media, but their compositions are highly variable. Our goal is to make growing media more sustainable by optimizing the selection of local and sustainable alternatives for peat. Different batches of COM and MR were incubated to assess the microbial activity based on (1) the N drawdown risk, (2) the C mineralization and (3) the inoculation efficiency of a commercially available biocontrol fungus. The various batches were characterized based on biochemical, chemical (pH, available and total nutrients) and microbiological biomass analysis. COM and MR were scored based on chemical or stability characteristics to assess their suitability to replace peat, lime and fertilizers in growing media. This score allowed for a clear differentiation between the materials; MR received higher scores on average than COM. Five composts were further tested for the effect of storage after blending with an acidic MR, acidification with elemental S, or removal of the finer fraction. One batch of chopped soft rush was acidified with elemental S. Blending and acidification were the most effective treatments as they resulted in a clear increase of the suitability score.

Renewable P sources: P use efficiency of digestate, processed animal manure, compost, biochar and struvite.

In the attempt to close nutrient cycles, organic fertilizers and soil improvers are getting interest as renewable P sources for crops. However, both the P availability of these compounds for crops and the underlying mechanisms are not fully understood. In this study composts (n = 8), biochars (n = 5), animal manure and processed animal manure (n = 13), digestates and processed digestates (n = 15) and blends of digestates with compost/animal manure/mineral fertilizers (n = 15) were analyzed for chemical composition, organic matter stability and P use efficiency (PUE). Biodegradability (=holocellulose/lignin ratio) proved to be a good indicator for organic matter stability and can successfully replace time-consuming incubation experiments in standard analyses of organic fertilizers. The PUE of digestates, struvites, animal manure products and blends of digestate with compost/animal manure/mineral fertilizers was determined by the NH4+-N, Mg and Fe content of the organic fertilizers. The PUE can be predicted by PUE = 61.34 + 8.59*NH4+-N/P + 42.25*Mg/P - 8.09*Fe/P (R2 = 0.71). As increasing amounts of NH4+-N and Mg stimulate the formation of soluble struvite crystals, increasing PUE is explained by an increasing amount of P as struvite. The PUE of biochars and composts was determined by the Ca/P and Al content of the organic fertilizers. Here, PUE can be predicted by PUE = 88.87-1.07*Ca/P + 6.08*Al/P (R2 = 0.93). As increasing amounts of Ca stimulate the formation of highly stable apatite crystals, increasing PUE is explained by an increasing amount of P in the form of apatite.

Daring to be differential: metabarcoding analysis of soil and plant-related microbial communities using amplicon sequence variants and operational taxonomical units.

BACKGROUND: Microorganisms are not only indispensable to ecosystem functioning, they are also keystones for emerging technologies. In the last 15 years, the number of studies on environmental microbial communities has increased exponentially due to advances in sequencing technologies, but the large amount of data generated remains difficult to analyze and interpret. Recently, metabarcoding analysis has shifted from clustering reads using Operational Taxonomical Units (OTUs) to Amplicon Sequence Variants (ASVs). Differences between these methods can seriously affect the biological interpretation of metabarcoding data, especially in ecosystems with high microbial diversity, as the methods are benchmarked based on low diversity datasets. RESULTS: In this work we have thoroughly examined the differences in community diversity, structure, and complexity between the OTU and ASV methods. We have examined culture-based mock and simulated datasets as well as soil- and plant-associated bacterial and fungal environmental communities. Four key findings were revealed. First, analysis of microbial datasets at family level guaranteed both consistency and adequate coverage when using either method. Second, the performance of both methods used are related to community diversity and sample sequencing depth. Third, differences in the method used affected sample diversity and number of detected differentially abundant families upon treatment; this may lead researchers to draw different biological conclusions. Fourth, the observed differences can mostly be attributed to low abundant (relative abundance < 0.1%) families, thus extra care is recommended when studying rare species using metabarcoding. The ASV method used outperformed the adopted OTU method concerning community diversity, especially for fungus-related sequences, but only when the sequencing depth was sufficient to capture the community complexity. CONCLUSIONS: Investigation of metabarcoding data should be done with care. Correct biological interpretation depends on several factors, including in-depth sequencing of the samples, choice of the most appropriate filtering strategy for the specific research goal, and use of family level for data clustering.

Introduction of a natural resource balance indicator to assess soil organic carbon management: Agricultural Biomass Productivity Benefit.

The rising demand for feed and food has put an increasing pressure on agriculture, with agricultural intensification as a direct response. Notwithstanding the higher crop productivity, intensive agriculture management entails many adverse environmental impacts. Worldwide, soil organic carbon (SOC) decline is hereby considered as a main danger which affects soil fertility and productivity. The life cycle perspective helps to get a holistic overview when evaluating the environmental sustainability of agricultural systems, though the impact of farm management on soil quality aspects is often not integrated. In this paper, we introduce an indicator called Agricultural Biomass Productivity Benefit of SOC management (ABB_SOC), which, relying on natural resource consumption, enables to estimate the net effect of the efforts made to attain a better soil quality. Hereby the focus is put on SOC. First, we introduce a framework to describe the SOC trend due to farm management decisions. The extent to which remediation measures are required are used as a measure for the induced SOC losses. Next, ABB_SOC values are calculated as the balance between the natural resource consumption of the inputs (including remediation efforts) and the desired output of arable crop production systems. The models RothC and EU-Rotate_N are used to simulate the SOC evolution due to farm management and the response of the biomass productivity, respectively. The developed indicator is applied on several rotation systems in Flanders, comparing different remediation strategies. The indicator could be used as a base for a method to account for soil quality in life cycle analysis.

Trichoderma-Inoculated Miscanthus Straw Can Replace Peat in Strawberry Cultivation, with Beneficial Effects on Disease Control.

Peat based growing media are not ecologically sustainable and often fail to support biological control. Miscanthus straw was (1) tested to partially replace peat; and (2) pre-colonized with a Trichoderma strain to increase the biological control capacity of the growing media. In two strawberry pot trials (denoted as experiment I & II), extruded and non-extruded miscanthus straw, with or without pre-colonization with T. harzianum T22, was used to partially (20% v/v) replace peat. We tested the performance of each mixture by monitoring strawberry plant development, nutrient content in the leaves and growing media, sensitivity of the fruit to the fungal pathogen Botrytis cinerea, rhizosphere community and strawberry defense responses. N immobilization by miscanthus straw reduced strawberry growth and yield in experiment II but not in I. The pre-colonization of the straw with Trichoderma increased the post-harvest disease suppressiveness against B. cinerea and changed the rhizosphere fungal microbiome in both experiments. In addition, defense-related genes were induced in experiment II. The use of miscanthus straw in growing media will reduce the demand for peat and close resource loops. Successful pre-colonization of this straw with biological control fungi will optimize crop cultivation, requiring fewer pesticide applications, which will benefit the environment and human health.

Improving the product stability and fertilizer value of cattle slurry solid fraction through co-composting or co-ensiling.

Separating dairy cattle slurry in a liquid and solid fraction (SF) is gaining more interest, since it enables a more targeted use of both fractions. However, the valorization of the SF is limited on P-rich soils, due to its high P content, and the export or use as bedding material requires sanitation. Therefore, we investigated the influence of composting or ensiling the SF, whether or not mixed with bulking agents, on the product quality in terms of fertilizer value, sanitation and stability. Ensiling can be considered as a controlled storage method for conserving C and nutrients. Soil amendment with co-ensiled SF resulted in a higher N mineralization and crop growth compared to amendment of co-composted SF. Co-composting SF with structure-rich feedstock materials optimized the composting process and sanitation when compared with composting pure SF and did not increase the risk for extreme-heat-resistant spores of thermophilic aerobic spore-forming bacteria (X-TAS). Further, the composts contained more P per unit of fresh weight than the silages, beneficial for the export of the composted SF. The oxygen uptake rate was found to be less powerful to determine the stability of fresh, composted and ensiled SF.

Combining woody biomass for combustion with green waste composting: Effect of removal of woody biomass on compost quality.

The question was tackled on how the green waste compost industry can optimally apply the available biomass resources for producing both bioenergy by combustion of the woody fraction, and high quality soil improvers as renewable sources of carbon and nutrients. Compost trials with removal of woody biomass before or after composting were run at 9 compost facilities during 3 seasons to include seasonal variability of feedstock. The project focused on the changes in feedstock and the effect on the end product characteristics (both compost and recovered woody biomass) of this woody biomass removal. The season of collection during the year clearly affected the biochemical and chemical characteristics of feedstock, woody biomass and compost. On one hand the effect of removal of the woody fraction before composting did not significantly affect compost quality when compared to the scenario where the woody biomass was sieved from the compost at the end of the composting process. On the other hand, quality of the woody biomass was not strongly affected by extraction before or after composting. The holocellulose:lignin ratio was used in this study as an indicator for (a) the decomposition potential of the feedstock mixture and (b) to assess the stability of the composts at the end of the process. Higher microbial activity in green waste composts (indicated by higher oxygen consumption) and thus a lower compost stability resulted in higher N immobilization in the compost. Removal of woody biomass from the green waste before composting did not negatively affect the compost quality when more intensive composting was applied. The effect of removal of the woody fraction on the characteristics of the green waste feedstock and the extracted woody biomass is depending on the season of collection.

Environmental life cycle assessment of grain maize production: An analysis of factors causing variability.

To meet the growing demand, high yielding, but environmentally sustainable agricultural plant production systems are desired. Today, life cycle assessment (LCA) is increasingly used to assess the environmental impact of these agricultural systems. However, the impact results are very diverse due to management decisions or local natural conditions. The impact of grain maize is often generalized and an average is taken. Therefore, we studied variation in production systems. Four types of drivers for variability are distinguished: policy, farm management, year-to-year weather variation and innovation. For each driver, scenarios are elaborated using ReCiPe and CEENE (Cumulative Exergy Extraction from the Natural Environment) to assess the environmental footprint. Policy limits fertilisation levels in a soil-specific way. The resource consumption is lower for non-sandy soils than for sandy soils, but entails however more eutrophication. Farm management seems to have less influence on the environmental impact when considering the CEENE only. But farm management choices such as fertiliser type have a large effect on emission-related problems (e.g. eutrophication and acidification). In contrast, year-to-year weather variation results in large differences in the environmental footprint. The difference in impact results between favourable and poor environmental conditions amounts to 19% and 17% in terms of resources and emissions respectively, and irrigation clearly is an unfavourable environmental process. The best environmental performance is obtained by innovation as plant breeding results in a steadily increasing yield over 25 years. Finally, a comparison is made between grain maize production in Flanders and a generically applied dataset, based on Swiss practices. These very different results endorse the importance of using local data to conduct LCA of plant production systems. The results of this study show decision makers and farmers how they can improve the environmental performance of agricultural systems, and LCA practitioners are alerted to challenges due to variation.

Dynamics in the Strawberry Rhizosphere Microbiome in Response to Biochar and Botrytis cinerea Leaf Infection.

Adding biochar, the solid coproduct of biofuel production, to peat can enhance strawberry growth, and disease resistance against the airborne fungal pathogen Botrytis cinerea. Additionally, biochar can induce shifts in the strawberry rhizosphere microbiome. However, the moment that this biochar-mediated shift occurs in the rhizosphere is not known. Further, the effect of an above-ground infection on the strawberry rhizosphere microbiome is unknown. In the present study we established two experiments in which strawberry transplants (cv. Elsanta) were planted either in peat or in peat amended with 3% biochar. First, we established a time course experiment to measure the effect of biochar on the rhizosphere bacterial and fungal communities over time. In a second experiment, we inoculated the strawberry leaves with B. cinerea, and studied the impact of the infection on the rhizosphere bacterial community. The fungal rhizosphere community was stabilized after 1 week, except for the upcoming Auriculariales, whereas the bacterial community shifted till 6 weeks. An effect of the addition of biochar to the peat on the rhizosphere microbiome was solely measured for the bacterial community from week 6 of plant growth onwards. When scoring the plant development, biochar addition was associated with enhanced root formation, fruit production, and postharvest resistance of the fruits against B. cinerea. We hypothesize that the bacterial rhizosphere microbiome, but also biochar-mediated changes in chemical substrate composition could be involved in these events. Infection of the strawberry leaves with B. cinerea induced shifts in the bacterial rhizosphere community, with an increased bacterial richness. This disease-induced effect was not observed in the rhizospheres of the B. cinerea-infected plants grown in the biochar-amended peat. The results show that an above-ground infection has its effect on the strawberry rhizosphere microbiome, changing the bacterial interactions in the root-substrate interface. This infection effect on the bacterial rhizosphere microbiome seems to be comparable to, but less pronounced than the effect of biochar-addition to the peat. The biological meaning of these observations needs further research, but this study indicates that biochar and an above-ground pathogen attack help the plant to recruit rhizosphere microbes that may aid them in their plant growth and health.

Biochar amendment before or after composting affects compost quality and N losses, but not P plant uptake.

We investigated the use of biochar (10% on a dry weight basis) to improve the composting process and/or the compost quality by adding it to either the feedstock mixture or the mature compost. The addition of biochar to the feedstocks was essayed in a full scale trial using a mixture of green waste and the organic fraction of municipal solid waste. Addition of biochar to mature compost was performed in a medium scale experiment. The use of biochar, even in small amounts, changed the composting process and the properties of the end products. However these effects depended on the time of application. We observed a faster decomposition in the bio-oxidative phase and lower greenhouse gas emissions when biochar was added at the beginning of the composting process, and a reduction in readily available P when biochar was applied during compost storage. Biochar as a means to increase the C content of the compost was only effective during compost storage. The P fertilizer replacement value of the compost with and without biochar was tested in a plant trial with annual ryegrass. While there was a clear effect on readily available P concentrations in the compost, adding biochar to the feedstock or the compost did not affect the P fertilizer replacement value.

Organ- and species-specific accumulation of metals in two land snail species (Gastropoda, Pulmonata).

In order to evaluate the usefulness of terrestrial gastropods as bioindicators there is a need for studies that simultaneously compare (1) concentrations of metals in reference and polluted plots, (2) species within the same polluted habitat, (3) metal accumulation patterns in different organs and (4) metal accumulation patterns in relation to soil physicochemical properties. This study aims to assess metal accumulation patterns in two land snail species. Instead of analyzing an organism as a whole, investigating the partitioning of metals in different organs can provide information on the actual toxicological relevant fractions. Therefore, concentrations of Ag, Cd, Cr, Cu, Ni and Zn were examined in five different organs of Cepaea nemoralis, as well as in the foot and the body of Succinea putris. Snails were sampled at four polluted dredged sediment disposal localities and three adjacent less polluted reference plots situated along waterways in Flanders, Belgium. Due to the small size and problematic dissection of S. putris only the concentrations in the foot of both species could be compared. For this reason only, C. nemoralis can be described as a better bioindicator species that allows a far more detailed analysis of organ metal accumulation. This study showed that organs other than the digestive gland may be involved in the immobilization and detoxification of metals. Furthermore, pH, soil fractionation (clay %, silt %, sand %) and organic matter, correlate with metal accumulation in organs. However, most often the soil metal concentrations did not correlate with the concentrations found in snail organs. Metal concentrations in organs of both species (1) differed among polluted plots but rarely between polluted and reference plots within a locality, (2) were organ-specific (digestive gland>foot>albumen gland=spermoviduct=ovotestis), (3) were species-specific and (4) depended on the metal type (high Cd and Cu concentrations were observed in the digestive gland and foot respectively). Our study emphasizes that background metal levels should be taken into account when using invertebrates as bioindicators of metal contamination and that bioindicators may show substantial differences in accumulation patterns even if they have a highly comparable ecology.

Fertilizing soil with selenium fertilizers: impact on concentration, speciation, and bioaccessibility of selenium in leek (Allium ampeloprasum).

Leek was fertilized with sodium selenite (Na(2)SeO(3)) and sodium selenate (Na(2)SeO(4)) in a green house to assess the impact of selenium (Se) fertilization on Se uptake by the crop and its speciation in the crop. The bioaccessibility of Se in the Se-enriched leek was assessed using an in vitro extraction protocol mimicking the human gastrointestinal tract (stomach, small intestine, and colon). The lowest Se uptake was observed when Na(2)SeO(3) was used as a fertilizer, which results in a higher risk for Se accumulation in the soil on a longer term. When soil was amended with Na(2)SeO(4), 55 ± 5% of total Se in the leek occurred in an inorganic form, while only 21 ± 8% was inorganic when Na(2)SeO(3) was applied. Se-methylselenocysteine and selenomethione were the major organic species in both treatments. However, concentrations of Se-methylselenocysteine and γ-glutamyl-Se-methyl-selenocysteine, which were previously reported to induce positive health effects, were lower as compared to other Allium species. The majority of the Se in the leek was found to be bioaccessible in the stomach (around 60%) and small intestine (around 80%). However, a significant fraction also has good chances to reach the colon, where it seems to be taken up by the microbial community and may also induce positive health effects.

Influence of flooding and metal immobilising soil amendments on availability of metals for willows and earthworms in calcareous dredged sediment-derived soils.

Soil amendments previously shown to be effective in reducing metal bioavailability and/or mobility in calcareous metal-polluted soils were tested on a calcareous dredged sediment-derived soil with 26 mg Cd/kg dry soil, 2200 mg Cr/kg dry soil, 220 mg Pb/kg dry soil, and 3000 mg Zn/kg dry soil. The amendments were 5% modified aluminosilicate (AS), 10% w/w lignin, 1% w/w diammonium phosphate (DAP, (NH4)2HPO4), 1% w/w MnO, and 5% w/w CaSO4. In an additional treatment, the contaminated soil was submerged. Endpoints were metal uptake in Salix cinerea and Lumbricus terrestris, and effect on oxidation-reduction potential (ORP) in submerged soils. Results illustrated that the selected soil amendments were not effective in reducing ecological risk to vegetation or soil inhabiting invertebrates, as metal uptake in willows and earthworms did not significantly decrease following their application. Flooding the polluted soil resulted in metal uptake in S. cinerea comparable with concentrations for an uncontaminated soil.