Sample preparation, preservation, and storage for volatile fatty acid quantification in biogas plants.

Volatile fatty acids (VFA) represent short-chain fatty acids consisting of six or fewer carbon atoms that can be distilled at atmospheric pressure. In anaerobic digestion processes VFAs are of central importance for maintaining stable reactor performance and biogas production, are used as indicators for arising problems and are important process monitoring parameters. In the present study, sludge derived form a full-scale anaerobic digester of a wastewater treatment plant was spiked with formate, acetate, propionate, and butyrate in order to evaluate various commonly used techniques for VFA extraction, preservation, and storage. It was shown that VFA extraction after centrifugation warranted the highest recovery rates for spiked VFAs. Moreover, experiments clearly indicated the importance of a fast sample handling, including the necessity of immediate cooling of the samples. Chemical sample preservation within a narrow time frame or deep freezing emerged as an alternative to instant VFA extraction. Short-time storage of extracted VFA samples at + 4°C is an option for up to 7 days, for longer periods storage at -20°C was found to be applicable.

Mevalonate governs interdependency of ergosterol and siderophore biosyntheses in the fungal pathogen Aspergillus fumigatus.

Aspergillus fumigatus is the most common airborne fungal pathogen for humans. In this mold, iron starvation induces production of the siderophore triacetylfusarinine C (TAFC). Here we demonstrate a link between TAFC and ergosterol biosynthetic pathways, which are both critical for virulence and treatment of fungal infections. Consistent with mevalonate being a limiting prerequisite for TAFC biosynthesis, we observed increased expression of 3-hydroxy-3-methyl-glutaryl (HMG)-CoA reductase (Hmg1) under iron starvation, reduced TAFC biosynthesis following lovastatin-mediated Hmg1 inhibition, and increased TAFC biosynthesis following Hmg1 overexpression. We identified enzymes, the acyl-CoA ligase SidI and the enoyl-CoA hydratase SidH, linking biosynthesis of mevalonate and TAFC, deficiency of which under iron starvation impaired TAFC biosynthesis, growth, oxidative stress resistance, and murine virulence. Moreover, inactivation of these enzymes alleviated TAFC-derived biosynthetic demand for mevalonate, as evidenced by increased resistance to lovastatin. Concordant with bilateral demand for mevalonate, iron starvation decreased the ergosterol content and composition, a phenotype that is mitigated in TAFC-lacking mutants.