Claudia Preininger

Implementing “soil health” means sustainable management of agricultural soils, avoiding toxicities, and sensible use of resources to minimize waste. In this context, the use of plastic in agriculture in form of plastic products, the application of polymers and additives in fertilizers, and plastic input through littering and tyre wear demands our special attention. Uncertainty and open questions relating to effects of plastic and its degradation products such as microplastic (MP) on the soil environment, the soil biota, and human health partly result from the lack of robust and standardized detection and measurement methods. Also, environmental, economic, and societal problems around MPs in soil cannot be adequately addressed due to lack of coordination among the various relevant players and initiatives in research and policy. NETmicroplastic ( www.net-microplastic.eu ) responds to the need of connecting among a fragmented research & innovation and policy landscape by creating a community‐supported environment. The network fosters provision of solid data for science‐based impact assessment of MP in soil together with much‐needed technological innovations, including biodegradable alternatives to conventional plastic. Here, we reflect upon a number of action fields that are key to the NETmicroplastic initiative from small to large‐scale perspectives. In addition, we portray the overall awareness situation around MP in soil.

One of the most widely known biogenic amines is histamine, which plays an important role in the human immune system. Some people suffer from allergic reactions after a histamine-rich diet; this is called histamine intolerance. The aim of this work is to develop a quick and reliable method for the detection and quantification of histamine in food, based on an electrochemical approach. In presence of biogenic amines, a reduction cascade induces a current at the working electrode. Prior to chronoamperometric measurements, Finite Elemente simulations were performed. The results are presented in this work.

Damages of the (agro)ecosystem by extensive use of chemical fertilizers and pesticides, the global dying of bee populations possibly linked to pesticide spraying, and stricter regulations for pesticide use together with successful use of microbials in IPM programs are pushing on the development and commercialization of new microbial products and a large and growing biostimulants and biocontrol market. This review focuses on microbial inoculants including bacteria, fungi, and viruses used as biostimulant or biocontrol agent for foliar application and covers all important steps from inoculant development to successful field application. Topics presented comprise typical spraying equipment including the importance of the spraying process and relating effects, furthermore formulation development including classification and adjuvants, and thirdly regulatory aspects as currently applied or under discussion. Microbial inoculants for foliar spray reported in scientific literature are summarized and contrasted with selected commercial products. Special attention is given to factors most important in microbial spray: (a) type of active ingredient (bacteria, fungi, viruses), (b) mode of action (ingestion, contact, competition), (c) interaction with the plant leaf surface, (d) droplet size in terms of microbe concentration and leaf coverage, and (e) environmental conditions during spraying. Finally, we want to emphasize that timely administration is of utmost importance for successful spraying and maximum efficacy. This might be supported by weather stations and disease/pest models as an important step towards precision farming.

A colorimetric microarray for the multiplexed detection of recurrence of bladder cancer including protein markers interleukin-8 (IL8), decorin (DCN), and vascular endothelial growth factor (VEGF) was established to enable easy and cheap read-out by a simple office scanner paving the way for quick therapy monitoring at doctors’ offices. The chip is based on the principle of a sandwich immunoassay and was optimized prior to multiplexing using IL8 as a model marker. Six different colorimetric assay formats were evaluated using a detection antibody (dAB) labeled with (I) gold (Au) nanoparticles (NPs), (II) carbon NPs, (III) oxidized carbon NPs, and a biotinylated dAB in combination with (IV) neutravidin–carbon, (V) streptavidin (strp)–gold, and (VI) strp–horseradish peroxidase (HRP). Assay Format (III) worked best for NP-based detection and showed a low background while the enzymatic approach, using 3,3′,5,5′-tetramethylbenzidine (TMB) substrate, led to the most intense signals with good reproducibility. Both assay formats showed consistent spot morphology as well as detection limits lower than 15 ng/L IL8 and were thus applied for the multiplexed detection of IL8, DCN, and VEGF in synthetic urine. Colorimetric detection in urine (1:3) yields reaction signals and measurement ranges well comparable with detection in the assay buffer, as well as excellent data reproducibility as indicated by the coefficient of variation (CV 5–9%).

This study describes the development and the challenges in the development of an on-chip immunoassay for histamine using commercially available antibodies. Histamine can be used as an indicator of food freshness and quality, but it is also a relevant marker in clinical diagnostics. Due to its low molecular weight, simple structure and thus low immunogenicity production of high specificity and affinity antibodies is difficult. From six commercial anti-histamine antibodies tested, only two bound the histamine free in the solution. A fluorescent on-chip immunoassay for histamine was established with a dynamic range of 8–111 µg/mL using polyclonal anti-histamine antibody H7403 from Sigma (Mendota Heights, MN, USA). The anti-histamine antibodies described and used in published literature are thoroughly reviewed and the quality of commercial antibodies and their traceability and quality issues are highlighted and extensively discussed.

Summary: The application of beneficial, plant‐associated microorganisms is a sustainable approach to improving crop performance in agriculture. However, microbial inoculants are often susceptible to prolonged periods of storage and deleterious environmental factors, which negatively impact their viability and ultimately limit efficacy in the field. This particularly concerns non‐sporulating bacteria. To overcome this challenge, the availability of protective formulations is crucial. Numerous parameters influence the viability of microbial cells, with drying procedures generally being among the most critical ones. Thus, technological advances to attenuate the desiccation stress imposed on living cells are key to successful formulation development. In this review, we discuss the core aspects important to consider when aiming at high cell viability of non‐sporulating bacteria to be applied as microbial inoculants in agriculture. We elaborate the suitability of commonly applied drying methods (freeze‐drying, vacuum‐drying, spray‐drying, fluidized bed‐drying, air‐drying) and potential measures to prevent cell damage from desiccation (externally applied protectants, stress pre‐conditioning, triggering of exopolysaccharide secretion, ‘helper’ strains). Furthermore, we point out methods for assessing bacterial viability, such as colony counting, spectrophotometry, microcalorimetry, flow cytometry and viability qPCR . Choosing appropriate technologies for maintenance of cell viability and evaluation thereof will render formulation development more efficient. This in turn will aid in utilizing the vast potential of promising, plant beneficial bacteria as sustainable alternatives to standard agrochemicals.

Herein, we present a workflow for the development of talc-based microbial inoculants for foliar spray consisting of four steps. These include together with decision-making criteria (1) the selection of additives based on their capability to wet juvenile maize leaves, (2) their adhesion on the plant, (3) their interaction with the biological systems, and (4) the choice of thickener for good dispersion stability. In total, 29 additives including polysaccharides and proteins, polyols, glycosides, oils, waxes, and surfactants (e.g., chitosan, gelatin, glycerol, saponin, castor oil, polyethylene, rhamnolipid) were evaluated. Contact angle and spreading index measurements revealed that the use of 5% Geloil, 1% rhamnolipid, or suitable combinations of Geloil + rhamnolipid and Nurture Yield S 2002 + rhamnolipid enhanced wetting of hydrophobic maize leaves and adherence, similarly to the commercial wetting agents recommended for plant protection 1% Prev B2 and 1% Trifolio S Forte. Interaction of additives with biological systems was based on biocompatibility and phytotoxicity assays, and cell viability monitoring using the endophytic Gram-negative bacterium Paraburkholderia phytofirmans PsJN. Results from biocompatibility assays indicated that in contrast to rhamnolipid and Prev B2 Geloil, Nurture Yield S 2002 and Trifolio S Forte fully supported bacterial growth within a concentration range of 1 to 5%. Dose-dependent phytotoxicity was observed in plants treated with rhamnolipid. Most efficient formulation was composed of PsJN, talc, xanthan, and Geloil. Beyond that, the proposed workflow is expected to generally provide guidance for the development of spray formulations and help other researchers to optimize their choices in this area.

The fluorescence-based multi-analyte chip platform for the analysis of estrogenic and anti-estrogenic substances is a new in vitro tool for the high throughput screening of environmental samples. In contrast to existing tools, the chip investigates the complex action of xenoestrogens in a human cell model by characterizing protein expression. It allows for the quantification of 10 proteins secreted by MCF-7 cells, representing various biological and pathological endpoints of endocrine action and distinguishing between estrogen- and anti-estrogen-dependent secretion of proteins. Distinct protein secretion patterns of the cancer cell line after exposure to known estrogen receptor agonists ß-estradiol, bisphenol A, genistein, and nonylphenol as well as antagonists fulvestrant and tamoxifen demonstrate the potential of the chip. Stimulation of cells with Interleukin-1ß shifts concentrations of low abundant biomarkers towards the working range of the chip. In the non-stimulated cell culture, Matrix Metalloproteinase 9 (MMP-9) and Vascular Endothelial Growth Factor (VEGF) show differences upon treatment with antagonists and agonists of the estrogen receptor. In stimulated MCF-7 cells challenged with receptor agonists secretion of Monocyte Chemoattractant Protein (MCP-1), Interleukin-6 (IL-6), Rantes, and Interleukin-8 (IL-8) significantly decreases. In parallel, the proliferating effect of endocrine-disrupting substances in MCF-7 cells is assessed in a proliferation assay based on resazurin. Using ethanol as a solvent for test substances increases the background of proliferation and secretion experiments, while using dimethyl sulfoxide (DMSO) does not show any adverse effects. The role of the selected biomarkers in different physiological processes such as cell development, reproduction, cancer, and metabolic syndrome makes the chip an excellent tool for either indicating endocrine-disrupting effects in food and environmental samples, or for screening the effect of xenoestrogens on a cellular and molecular level.