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SummaryThe 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.

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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.

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Four different materials (two ab initio synthesized polyurethanes; ready-to-use: Epon1002F and poly(vinyl alcohol)/N-methyl-4(4'-formylstyryl)pyridinium methosulfate acetal) for the generation of Escherichia coli surface imprints are compared in this work. The use of commercially available, ready-to-use materials instead of self-synthesized polymers represents an innovative and convenient way of molecular imprint fabrication. This was herein investigated for large, biological templates. Fully synthesized imprint materials (polyurethanes) were developed and optimized regarding their OH excess and the use of catalyst in the polymerization reaction. No to low OH excess (0-10%) and a noncatalyzed synthesis were determined to be superior for the imprinting of the Gram-negative bacteria. Imprints were characterized using atomic force microscopy, with Epon1002F yielding the most distinguished imprints, along with a smooth surface. The imprints were afterward tested as plastic antibody coatings in a mass-sensitive quartz-crystal microbalance measurement. Dilutions of E. coli suspensions, down to a limit of detection of 1.4 × 10 CFU/mL, were successfully measured. Best results were obtained with Epon1002F and self-synthesized, stoichiometric polyurethane. Since ready-to-use Epon1002F was superior in terms of signal intensities and sensitivity, it can advantageously replace self-synthesized polymers for the generation of imprinted sensor surfaces. Easy day-to-day reproducibility and further shortening of imprint fabrication time are other advantages of employing the ready-to-use material instead of conventionally synthesized polymers.

Neonatal sepsis is still a leading cause of death among newborns. Therefore a protein-microarray for point-of-care testing that simultaneously quantifies the sepsis associated serum proteins IL-6, IL-8, IL-10, TNF alpha, S-100, PCT, E-Selectin, CRP and Neopterin has been developed. The chip works with only a 4 μL patient serum sample and hence minimizes excessive blood withdrawal from newborns. The 4 μL patient samples are diluted with 36 μL assay buffer and distributed to four slides for repetitive measurements. Streptavidin coated magnetic particles that act as distinct stirring detection components are added, not only to stir the sample, but also to detect antibody antigen binding events. We demonstrate that the test is complete within 2.5 h using a single step assay. S-100 conjugated to BSA is spotted in increasing concentrations to create an internal calibration. The presented low volume protein-chip fulfills the requirements of point-of-care testing for accurate and repeatable (CV < 14%) quantification of serum proteins for the diagnosis of neonatal sepsis.

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