The iron uptake mechanisms based on siderophore synthesis used by the fish pathogen Aeromonas salmonicida subsp. salmonicida are still not completely understood, and the precise structure of the siderophore(s) is unknown. The analysis of genome sequences revealed that this bacterium possesses two gene clusters putatively involved in the synthesis of siderophores. One cluster is a candidate to encode the synthesis of acinetobactin, the siderophore of the human pathogen Acinetobacter baumannii, while the second cluster shows high similarity to the genes encoding amonabactin synthesis in Aeromonas hydrophila. Using a combination of genomic analysis, mutagenesis, biological assays, chemical purification, and structural determination procedures, here we demonstrate that most A. salmonicida subsp. salmonicida strains produce simultaneously the two siderophores, acinetobactin and amonabactin. Interestingly, the synthesis of both siderophores relies on a single copy of the genes encoding the synthesis of the catechol moiety (2,3-dihydroxybenzoic acid) and on one encoding a phosphopantetheinyl transferase. These genes are present only in the amonabactin cluster, and a single mutation in any of them abolishes production of both siderophores. We could also demonstrate that some strains, isolated from fish raised in seawater, produce only acinetobactin since they present a deletion in the amonabactin biosynthesis gene amoG. Our study represents the first evidence of simultaneous production of acinetobactin and amonabactin by a bacterial pathogen and reveals the plasticity of bacterial genomes and biosynthetic pathways. The fact that the same siderophore is produced by unrelated pathogens highlights the importance of these systems and their interchangeability between different bacteria.
The Andean church of San Andrés de Pachama is located in the highland of the northernmost of Chile, near the limit with Bolivia and next to the Ruta de la Plata. This commercial route contributed in the past to the transport and commerce of various raw materials, such as silver, from the Andean mountains region to the Pacific Ocean coast and then to the European market. The walls inside the church are decorated with paintings from the end of the eighteenth century that reproduce religious motivs together with flowers, fruits, and birds. In this study, micro samples taken from one of the mural paintings have been analysed to acquire information on the artistic materials and the painting technique previous to the restoration of the paintings. Analysis by micro-Raman spectroscopy complemented with scanning electron microscopy-energy dispersive X-ray spectroscopy, attenuated total reflection infrared spectroscopy, X-ray diffraction, and high-performance liquid chromatography with diode-array detection allowed the identification of orpiment, vermilion, indigo, smalt, antlerite, hematite, carmine lake, and wood charcoal as pigments as well as gypsum as the ground layer. Lipidic and proteinaceous materials extracted from the microsamples were identified by gas chromatography coupled to mass spectrometry and indicated the use of a mixture of egg and siccative oil as binders and a secco painting technique involving animal glue as the plaster primer. Smalt and the prized cochineal lake are reported for the first time in an Andean colonial mural painting.
The series of the Sibyls from the church of San Pedro Telmo is one of the most important groups of paintings of Argentine colonial art. Ten of the paintings were performed in the 18th century, while those corresponding to the Delphic and Tiburtine Sibyls were painted in 1864 during the first restoration of the series in Buenos Aires. There is a controversy regarding the origin of this remarkable group of paintings pointing to an Andean workshop or a Spanish source. In this study, Raman microscopy in combination with scanning electron microscope energy-dispersive X-ray spectrometer analysis and HPLC has successfully identified the materials employed in the manufacture of the paintings. The Raman analysis revealed the presence of a mixture of barium sulfate, lead white, and calcite in the preparation layers of the 19th century Sibyls in contrast to the clayish ground layer in the 18th century paintings. Traditional pigments such as vermilion, lead white, orpiment, indigo, ultramarine blue, and Prussian blue were readily identified by Raman microscopy. The presence of a madder lake in one of the 18th century Sibyls strongly suggests their Spanish origin in accordance with historical data. The identification of synthetic ultramarine blue in the Tiburtine painting is the first report of this blue pigment in Argentine colonial art.
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