Hahn, Martin W.

Abstract Sil.va.ni.grel'la. N.L. fem. dim. n. Silvanigrella named after Silva nigra the Latin geographic name of the Schwarzwald (Black Forest) mountains located in the south‐west of Germany. Proteobacteria / Oligoflexia / Silvanigrellales / Silvanigrellaceae / Silvanigrella The genus Silvanigrella accommodates heterotrophic freshwater bacteria isolated from the water column of circumneutral or slightly acidic (pH 5–7) lakes and ponds with low ion concentrations (conductivity < 30 μS/cm). So far, only the two species Silvanigrella aquatica and Silvanigrella paludirubra have been described. Cells of the type strains have in common red pigmentation, motility, and pleomorphic morphology. In both species, the cell morphology ranges from rod‐shaped cells to filamentous forms, the cell size is variable in length and width. The type strain of S. aquatica in addition forms, densely coiled spirals. The type strains of both species grow at temperatures above 30°C, but only the type strain of S. paludirubra grows at temperatures below 10°C. Growth occurred up to supplementary NaCl concentrations of 1.0–1.1% (w/v). Both type strains utilized various organic substrates including carbohydrates, amino acids, and short‐chain fatty acids; however, the substrate spectra of the two strains differed. The most abundant fatty acid was iso‐C 15:0 for both type strains, contributing 33% to the total fatty acid content. The major respiratory quinone of S. paludirubra is menaquinone MK‐8, while the quinones of S. aquatica remained unidentified. The type strains of S. aquatica and S. paludirubra were both characterized by medium‐sized genomes (3.5 and 3.9 Mb, respectively) with low G + C content (32.6 and 29.3 mol%, respectively). DNA G + C content (mol%) : 29–33 (genome sequencing). Type species : Silvanigrella aquatica Hahn et al. 2017 VP .

The actinobacterial strain 15G-AUS-rotT was isolated from an artificial pond located near Salzburg, Austria. The strain showed 16S rRNA gene sequence similarities of 98.7 % to Candidatus Aquiluna rubra and of 96.6 and 96.7 % to the two validly described species of the genus Rhodoluna . Phylogenetic reconstructions based on 16S rRNA gene sequences and genome-based on amino acid sequences of 118 single copy genes referred strain 15G-AUS-rotT to the family Microbacteriaceae and therein to the so-called subcluster Luna-1. The genome-based phylogenetic tree showed that the new strain represents a putative new genus. Cultures of strain 15G-AUS-rotT were light red pigmented and comprised very small, rod-shaped cells. They metabolized a broad variety of substrates. Major fatty acids (>10 %) of cells were iso-C16 : 0, antiso-C15 : 0 and iso-C14 : 0. The major respiratory quinone was MK-11 and a minor component was MK-10. The peptidoglycan structure belonged to an unusual B type. The closed genome sequence of the strain was very small (1.4 Mbp) and had a DNA G+C content of 54.8 mol%. An interesting feature was the presence of genes putatively encoding the complete light-driven proton pumping actinorhodopsin/retinal system, which were located at three different positions of the genome. Based on the characteristics of the strain, a new genus and a new species termed Aquiluna borgnonia is proposed for strain 15G-AUS-rotT (=DSM 107803T=JCM 32974T).

Filamentous bacteria frequently occurring in the pelagic zone of natural freshwater lakes and ponds were previously identified as being related to Haliscomenobacter hydrossis based upon their 16S rRNA gene sequences. These bacteria exhibit a specific morphology characterized by the formation of straight, stick-like filaments of variable length (5 to >100 μm) and quite stable, but narrow, width (0.25 to 0.35 μm). Bacteria with these morphological characteristics form a monophyletic but broad phylogenetic group with a maximal divergence of 16S rRNA gene sequences of 12.0 %. This monophyletic group consists of at least three monophyletic subclusters. H. hydrossis is affiliated to one of these subclusters and represents the sole recognized species affiliated to the broad monophyletic group. ‘Candidatus Haliscomenobacter calcifugiens' and ‘Candidatus Aquirestis calciphila’ are uncultured representatives of the other two subclusters and have 16S rRNA gene sequence dissimilarities of 5.4 % and 8.2 %, respectively, with the type strain of H. hydrossis. ‘Candidatus H. calcifugiens' and ‘Candidatus A. calciphila’ have a 16S rRNA gene sequence dissimilarity of 8.5 %. These large ribosomal divergences justify the classification of these environmentally important bacteria as a novel species and a new genus, respectively. Intensive attempts to cultivate these filamentous bacteria have resulted in the establishment of mixed cultures, however, attempts to establish pure cultures have failed.