Eunotia (Bacillariophyceae) from a subtropical stream adjacent to Iguaçu National Park, Brazil, with the proposition of a new species

Eunotia species from a stream located in the surrounding area of the Iguaçu National Park conservation unit were analyzed. Samples were monthly collected from September 2012 to August 2013. The periphytic material, manually obtained by squeezing aquatic macrophytes, was oxidized and analyzed under light and scanning electron microscopy. Twenty-one infrageneric taxa of Eunotia were identified, measured, and illustrated. Eunotia caniculoides sp. nov. is proposed as a new species, and eight taxa are new records for Paraná state.

Duganella lactea sp. nov., Duganella guangzhouensis sp. nov., Duganella flavida sp. nov. and Massilia rivuli sp. nov., isolated from a subtropical stream in PR China and proposal to reclassify Duganella ginsengisoli as Massilia ginsengisoli comb. nov.

Five Gram-stain-negative, catalase- and oxidase-positive, rod-shaped and motile strains (FT50WT, FT80WT, FT92WT, FT94W and FT135WT) were isolated from a subtropical stream in PR China. Comparisons based on 16S rRNA gene sequences showed that strains FT50WT, FT94W and FT135WT take strain Duganella sacchari Sac-22T, and strains FT80WT and FT92WT take strain Duganella ginsengisoli DCY83T as their closest neighbour in the phylogenetic trees, respectively. The G+C contents of strains FT50WT, FT80WT, FT92WT, FT94W and FT135WT were 63.3, 62.4, 62.8, 63.8 and 60.8 %, respectively. The reconstructed phylogenomic tree based on concatenated 92 core genes showed that strains FT50WT, FT80WT, FT94W and FT135WT clustered together with species of the genus Duganella , but strains FT92WT and D. ginsengisoli KCTC 42409T were located in the clades of the genus Massilia . The calculated pairwise average nucleotide identity (ANI) and digital DNA–DNA hybridization (dDDH) values among strains FT50WT, FT80WT, FT92WT, FT94W, FT135WT and related strains were in the ranges of 75.6–87.8% and 20.3–33.8% except that the values between strains FT50WT and FT94W were 98.7 and 89.2%, respectively. The respiratory quinone of these five strains was Q-8. The major fatty acids were C16 : 1 ω7c, C16 : 0, C18 : 1 ω7c and C12 : 0. The polar lipids included phosphatidylethanolamine, phosphatidylglycerol and one unidentified phospholipid. Considering the distinct phylogenetic relationships of D. ginsengisoli with species of the genus Massilia in the phylogenomic tree, it was reasonable to transfer D. ginsengisoli to the genus Massilia as Massilia ginsengisoli comb. nov. Combining the results of phylogenomic analysis, ANI and dDDH data, and a range of physiological and biochemical characteristics together, strains FT50WT and FT94W should belong to the same species and be assigned to genus Duganella with strains FT80WT and FT135WT together, and strain FT92WT should be assigned to the genus Massilia , for which the names Duganella lactea sp. nov. (type strain FT50WT=GDMCC 1.1674T=KACC 21466T), Duganella guangzhouensis sp. nov. (FT80WT=GDMCC 1.1678T=KACC 21470T), Duganella flavida sp. nov. (FT135WT=GDMCC 1.1745T=KACC 21659T) and Massilia rivuli sp. nov. (FT92WT=GDMCC 1.1682T=KACC 21474T) are proposed.

Rugamonas aquatica sp. nov. and Rugamonas rivuli sp. nov., isolated from a subtropical stream in PR China

Two Gram-stain-negative, catalase- and oxidase-positive, rod-shaped, motile strains (FT29WT and FT103WT) were isolated from a subtropical stream in PR China. Comparisons based on 16S rRNA gene sequences showed that strains FT29WT and FT103WT showed 98.7 and 98.6 % 16S rRNA gene sequence similarities to Rugamonas rubra CCM 3730T as their closest neighbour, respectively. The calculated pairwise OrthoANIu values between strain R. rubra CCM 3730T and strains FT29WT and FT103WT were all 81.4 %. The respiratory quinone of strains FT29WT and FT103WT was determined to be Q-8. The major fatty acids were C16 : 1 ω7c, C16 : 0 and C12 : 0. The polar lipids of strain FT103WT included phosphatidylethanolamine, phosphatidylglycerol and one unidentified phospholipid, but strain FT29WT did not contain phosphatidylglycerol. The genome sizes of strains FT29WT and FT103WT were 7.23 and 6.84 Mbp with G+C contents of 63.8 and 63.9 %, respectively. Although the pairwise OrthoANIu value between strains FT29WT and FT103WT was 95.2 % which located in the transition region of species demarcation, the dissimilarities in the aspects of phenotypic, biochemical and genotypic characteristics supported these two strains should belong to the different species within genus Rugamonas , for which the names Rugamonas aquatica sp. nov. (type strain FT29WT=GDMCC 1.1643T=KACC 21316T) and Rugamonas rivuli sp. nov. (type strain FT103WT=GDMCC 1.1685T=KACC 21477T) are proposed.

Janthinobacterium violaceinigrum sp. nov., Janthinobacterium aquaticum sp. nov. and Janthinobacterium rivuli sp. nov., isolated from a subtropical stream in China

Four strains assigned the names FT13WT, FT14W, FT58WT and FT68WT were isolated from a subtropical stream in PR China. All the strains were Gram-stain-negative, catalase- and oxidase-positive, rod-shaped and motile with flagella. Comparisons based on 16S rRNA gene sequences showed that strains FT13WT, FT14W, FT58WT and FT68WT belonged to genus Janthinobacterium and shared 16S rRNA gene similarities in the range of 98.8–99.7 % with Janthinobacterium lividum DSM 1522T, Janthinobacterium agaricidamnosum DSM 9628T and ‘Janthinobacterium svalbardensis JA-1', respectively. The calculated pairwise average nucleotide identity (ANI) values among the genomes of above seven strains were in the range of 79.0–92.2 %, except that the ANI value was 96.8 % between strain FT13WT and FT14W. The respiratory quinone of strains FT13WT, FT14W, FT58WT and FT68WT was determined to be Q-8. The major fatty acids were C16 : 1 ω7c, C16 : 0, C18 : 1 ω7c and C12 : 0. The polar lipids included phosphatidylethanolamine, phosphatidylglycerol and one unidentified phospholipid. The genome sizes of strains FT13WT, FT14W, FT58WT and FT68WT were 6.45, 6.38, 5.73 and 6.37 Mbp with G+C contents of 63.4, 63.7, 61.6 and 63.1 mol%, respectively. Combining phenotypic, biochemical, genotypic and ANI data, strain FT13WT and FT14W should belong to the same species. The four strains were considered to represent three novel species within genus Janthinobacterium , for which the names Janthinobacterium violaceinigrum sp. nov. (type strain FT13WT=GDMCC 1.1638T=KACC 21319T), Janthinobacterium aquaticum sp. nov. (FT58WT=GDMCC 1.1676T=KACC 21468T) and Janthinobacterium rivuli sp. nov. (FT68WT=GDMCC 1.1677T=KACC 21469T) are proposed.