Glycogen-accumulating organisms (GAOs) are found in enhanced biological phosphorus removal systems where they compete with polyphosphate-accumulating organisms for external carbon substrates. (13)C nuclear magnetic resonance ((13)C-NMR) was used to elucidate the metabolic pathways operating in an enriched GAO culture dominated by two known GAOs (81.2%). The experiments consisted of adding (13)C-acetate (labelled on position 1 or 2) to the culture under anaerobic conditions, and operating the culture through a cycle consisting of an anaerobic, an aerobic and a further anaerobic phase. The carbon transformations over the cycle were monitored using in vivo(13)C-NMR. The two-carbon moieties in hydroxybutyrate and hydroxyvalerate were derived from acetate, while the propionyl precursor of hydroxyvalerate was primarily derived from glycogen, with only a small fraction originating from acetate. Comparison of the labelling patterns in hydroxyvalerate at the end of the first and the second anaerobic periods in pulse experiments with 2-(13)C-acetate showed that the Entner-Doudoroff (ED) pathway was used for the breakdown of glycogen. This conclusion was further supported by the labelling pattern on glycogen observed in the pulse experiments with 1-(13)C-acetate, which can only be explained by the operation of ED with recycling of pyruvate and glyceraldehyde 3-phosphate via gluconeogenesis. The activity of the ED pathway for glycogen degradation by GAOs is demonstrated here for the first time. In addition, the decarboxylating part of the tricarboxylic acid cycle was confirmed to operate also under anaerobic conditions.
The Brown Hairstreak (Thecla betulae L.) is one of the least observed butterflies of the Palaearctic region, even though its distribution spans from Portugal in the west, to Russia and Korea in the far east. Adults are arboreal and seldom descend to ground level. As a result, this species is mostly monitored via the detection of eggs on the food plant during wintertime. In the Iberian Peninsula, this species was largely unknown until very recently, but a recent burst of regional studies in Spain has begun bridging this gap. However, their focused nature and a still incomplete knowledge on T. betulae in Portugal promoted the need for an integrative study at the Iberian scale.
Here, we carried out a full literature review on the distribution, ecology and behaviour of T. betulae in Portugal and Spain. Complemented with field work in Portugal, we revealed an almost continuous distribution in the northern third of Iberia, whilst populations further south are mostly mountain-bound. In order to help with future discovery of new populations, we built a species-distribution model relating its occurrence with bioclimatic variables. This model accurately explains the current known occupation of the territory and highlights other areas where the species may potentially be found. Finally, we found evidence of a broadening of the species’ niche through the local use of an hitherto unknown food plant. This study sets a new knowledge baseline for future works and conservation of T. betulae through southern Europe.
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