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Bacterial infections are a global threat to human health1. Especially nosocomial infections with multidrug-resistant Gram-negative bacteria challenge public health systems and endanger successful treatment of patients in many modern medicine applications as e.g. organ transplant. In the last three decades classical approaches failed to find novel chemical scaffolds suitable for the development of antibiotics that reach the market and caused the modern antibiotics crisis1.
Insect microbiomes produce potent antibiotics with low toxicity to eukaryotic cells to defend their hosts against entomopathogenic microorganisms2. In order to exploit insect microbiomes efficiently and to identify novel antibiotics producing bacteria, we implemented an ultra-high throughput Microfluidics/FACS (fluorescence-associated cell sorting) cultivation and screening pipeline. Here we present the technical aspects of our pipeline including downstream processing of identified producer strains and its suitability for antibiotics discovery.
Bacterial infections are a global threat to human health1. Especially nosocomial infections with multidrug-resistant Gram-negative bacteria challenge public health systems and endanger successful treatment of patients in various modern medicine applications as e.g. organ transplant. In the last three decades classical approaches failed to find novel chemical scaffolds suitable for the development of antibiotics.
Insect microbiomes produce potent antibiotics with low toxicity to eukaryotic cells to defend their hosts against entomopathogenic microorganisms2. In order to exploit insect microbiomes efficiently and to identify novel antibiotic producing bacteria, we implemented an high throughput Microfluidics/FACS (fluorescence-associated cell sorting) cultivation and screening pipeline. Here, we present the technical aspects of our pipeline including downstream processing. This includes genomic fingerprinting of identified producer strains, metabolomic analysis and isolation of natural products.
Sol-gel synthesis was used to prepare nanoparticles of MgF2 and CaF2. These nanoparticles were tested for their application in wood protection. In comparison to control samples, wood specimens treated with MgF2 and CaF2 showed lower mass loss under exposure to termites and fungi. Samples treated with MgF2 performed better at protecting wood from cellulose degradation when exposed to termites. Conversely, in the presence of fungi, mass loss in treated samples was similar irrespective of the chemical composition (MgF2 and CaF2) of the treatment. A possible explanation for this difference in results between termites and fungi could be their different mechanism of cellulose hydrolysis. Thus, future wood preservatives would need to be customized according to their specific application.
Alkaline earth metal fluoride nanoparticles have been investigated for application in wood protection. Sols of MgF2 and CaF2 were synthesized and their efficacy was tested against fungi and termites (Rehmer 2016, Krahl et al. 2016). The sols were characterized by XRD and SEM. The wood specimens were vacuum impregnated with nanoparticles and then exposed to fungi and termites according to EU certified test conditions. Our results show that wood impregnated with metal fluoride nanoparticles significantly reduce cellulose hydrolysis by fungi and termites. The wood samples were exposed to brown-rot fungi; Coniophora puteana and Poria placenta. Between the two fungi, the overall mass lost due to fungal degradation was lower for treated (MgF2 and CaF2) wood samples exposed to Coniophora puteana. Thus, the metal fluoride nanoparticles impregnated in the wood samples were more efficient in reducing cellulose degradation from Coniophora puteana than from Poria placenta. However the mass loss in samples treated with MgF2 was similar to those treated with CaF2, irrespective of type of fungi. Therefore, it is likely that fungal degradation in treated samples was dependent on the biocidal action of fluorides rather than on the differences in chemical and physical properties of MgF2 and CaF2, respectively. Conversely, for termite exposure, wood samples treated with MgF2 had lower cellulose degradation compared to those treated with CaF2. A possible explanation for this difference in results could be fungi and termites use separate mechanisms for cellulose hydrolysis which will be further investigated. Future experiments include testing the leaching potential of MgF2 and CaF2 nanoparticles from wood. The results from the leaching experiment will test if metal fluoride nanoparticles can provide long-term and environmentally safe protection to wood.