Valediction: descriptions of novel prokaryotic taxa published in Antonie van Leeuwenhoek —change in editorial policy and a signpost to the future?

Streptomyces corynorhini sp. nov., isolated from Townsend’s big-eared bats ( Corynorhinus townsendii ) Abstract Four bacterial strains, with the capability of inhibiting Pseudogymnoascus destructans, the causative agent of white-nose syndrome, were isolated from male Townsend’s big-eared bats (Corynorhinus townsendii, Family: Vespertilionidae) in New Mexico. Isolates AC161, AC162, AC208, and AC230T were characterised as a novel clade using morphological, phenotypic and phylogenetic analysis. A draft genome of the type strain was completed to determine its taxonomy and secondary metabolite biosynthetic potential. Multi-locus sequence analysis nests AC230T with neighbours Streptomyces scopuliridis (NRRL B-24574T), Streptomyces lushanensis (NRRL B-24994T), Streptomyces odonnellii (NRRL B-24891T) and Streptomyces niveus (NRRL 2466T). Further phylogenetic analysis showed the MLSA distances between AC230T and its near neighbours are much greater than the generally accepted threshold (> 0.007) for bacterial species delineation. DNA–DNA relatedness between AC230T and its near neighbours ranged between 25.7 ± 2.1 and 29.9 ± 2.4%. The DNA G+C content of the genomic DNA of the type strain is 71.7 mol%. Isolate AC230T presents a white to ivory hue on most ISP media and its micromorphology exhibits ovoid spores with smooth surfaces in flexuous chains. Based on our study of AC230T, the strain warrants the assignment to a novel species, for which the name Streptomyces corynorhini sp. nov. is proposed. The type strain is AC230T (= JCM 33171T, = ATCC TSD155T).

Sphingobium chungangianum sp. nov., isolated from rhizosphere of Pinus koraiensis Abstract A novel Gram-staining negative, yellow-pigmented, non-motile, aerobic and rod-shaped bacterium, designated MAH-11T, was isolated from rhizosphere of Pinus koraiensis and was characterised by using a polyphasic taxonomic approach. The colonies were smooth, circular and 0.3–1.0 mm in diameter when grown on R2A agar for 3 days. The strain was positive for both catalase and oxidase tests. Optimum growth temperature and pH were 28–30 °C and 7.0, respectively. Cell growth occurs on R2A agar, nutrient agar, Luria–Bertani agar and tryptone soya agar but not on MacConkey agar. The novel strain was found to be able to hydrolyse esculin but not casein, gelatin, starch, l-tyrosine, DNA, l-arginine, urea, Tween 20 and Tween 80. On the basis of 16S rRNA gene sequence analysis, strain MAH-11T belongs to the genus Sphingobium and is closely related to Sphingobium quisquiliarum P25T (98.1%), Sphingobium vermicomposti VC-230T (97.8%), Sphingobium mellinum WI4T (97.5%), Sphingobium barthaii KK22T(97.2%) and Sphingobium fuliginis TKPT (97.2%). In DNA–DNA hybridization tests, the DNA relatedness values between strain MAH-11T and its close phylogenetic neighbors were below 45.0%. The DNA G+C content was 64.5 mol% and the predominant respiratory quinone was identified as ubiquinone-10. The major cellular fatty acids were summed feature 8 (C18:1ω7c and/or C18:1ω6c), summed feature 3 (C16:1ω7c and/or C16:1ω6c) and C16:0. The DNA–DNA hybridization results in combination with chemotaxonomic and physiological data demonstrated that strain MAH-11T represents a novel species within the genus Sphingobium, for which the name Sphingobium chungangianum is proposed. The type strain is MAH-11T (= KACC 19836T = CGMCC 1.13749T).

Haloterrigena salifodinae sp. nov., an extremely halophilic archaeon isolated from a subterranean rock salt Abstract A novel extremely halophilic strain, designated ZY19T, was isolated from a rock salt sample from Yunnan salt mine, PR China. Strain ZY19T is neutrophilic, non-motile and requires at least 10% (w/v) NaCl for growth. Optimal growth is observed at 20–25% (w/v) NaCl, pH 7.5–8.0 and 42 °C. Mg2+ is not required for growth. The cells do not lyse in distilled water. On the basis of 16S rRNA gene sequence analysis, strain ZY19T belongs to the genus Haloterrigena (Htg.) and is closely related to Haloterrigena salina XH-65T(98.5% sequence similarity) and Haloterrigena turkmenica DSM 5511T (97.9%). Phylogenetic and phylogenomic analysis showed that strain ZY19T clusters with the species Htg. salina and Htg. turkmenica forming an independent clade separated from other members of the genus. The value of genomic average nucleotide identity (ANI) between strains ZY19T and its close relative, Htg. salina XH-65T was 94.2%. DNA–DNA relatedness between strains ZY19T and Htg. salina XH-65T revealed by in silico DNA–DNA hybridization (DDH) was 56.3%. Both the ANI value and the degree of in silico DDH are below the accepted threshold for members of the same species. The major polar lipids were found to consist of phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, sulfated diglycosyl-diether-1 and mannose-2, 6-disulfate (1 → 2)-glucose glycerol diether. The genomic DNA G+C content was determined to be 64.5 mol%. Based on the results of the phenotypic, chemotaxonomic, genetic similarity and inferred phylogeny, strain ZY19T is distinct from other validly named species and thus represents a novel species within the genus Haloterrigena, for which the name Haloterrigena salifodinae sp. nov. is proposed. The type strain is ZY19T (=CGMCC 1.16114T=NBRC 112981T).

Paraburkholderia strydomiana sp. nov. and Paraburkholderia steynii sp. nov.: rhizobial symbionts of the fynbos legume Hypocalyptus sophoroides Abstract Twelve nodulating Paraburkholderia strains isolated from indigenous South African fynbos legume Hypocalyptus sophoroides were investigated to determine their taxonomic status. Genealogical concordance analysis, based on six loci (16S rRNA, atpD, recA, rpoB, lepA and gltB), revealed that they separate into two consistent and exclusive groups. Average nucleotide identity and DNA–DNA hybridisation comparisons indicated that they were sufficiently divergent from their closest known phylogenetic relatives (Paraburkholderia caledonica and Paraburkholderia terrae, respectively) to be regarded as novel species. This was also supported by the results of fatty acid analysis and metabolic characterisation. For these two isolate groups, we accordingly propose the new species Paraburkholderia strydomiana sp. nov. with WK1.1fT (= LMG 28731T = SARCC1213T) as its type strain and Paraburkholderia steynii sp. nov. with HC1.1baT (= LMG 28730T = SARCC696T) as its type strain. Our data thus showed that H. sophoroidesmay be considered a promiscuous symbiotic partner due to its ability to associate with multiple species of Paraburkholderia.

A putative novel Methylobacter member (KRF1) from the globally important Methylobacter clade 2: cultivation and salient draft genome features Abstract Methane oxidation by methanotrophs is a very important environmental process in the mitigation of methane. Methylobacter (Mtb.) clade 2 members have been reported as dominant methane oxidisers in soils and sediments worldwide. We enriched and purified a methanotroph from a tropical rice field soil sample from India. The highly enriched culture showed the presence of motile, long and thick rods (3–5 µm × 0.9–1.2 µm) and minor presence of short, thin rods. The culture was purified on agarose medium and formed yellow colonies which showed the presence of only thick and long rods, henceforth termed as strain KRF1. Based on 16S rRNA gene sequence analysis, strain KRF1 shows close phylogenetic affiliation to Methylobacter tundripaludum SV96T (98.6% similarity). Due to the taxonomic novelty, and being the first member of Mtb. related to Mtb. tundripaludum from the tropics, the draft genome was sequenced. From the blastx analysis of the contigs, it was clear that the culture still had contamination of another organism, a Methylophilus species. The data binned in two clear bins: Mtb. related contigs and Methylophilus-related contigs. The binned draft genome of KRF1 shows features including the typical pathways for methane metabolism, denitrification and the presence of molybdenum iron and vanadium-iron nitrogenase genes. KRF1 is phylogenetically distinct from the five strains of Mtb. tundripaludum including SV96T, Lake Washington strains and OWC strains, showing ~ 26% DDH and ~ 81% ANIb values and a unique position in a phylogenomic tree. Subsequently, KRF1 has been completely purified from its methylotrophic partner and a pure culture has been established and maintained in a WDCM approved culture collection, the MACS Collection of Microorganisms (as MCM 1471). KRF1 is thus the first cultured member of a putative novel species of Methylobacter clade 2 isolated from the tropics.

Lysobacter caseinilyticus , sp. nov., a casein hydrolyzing bacterium isolated from sea water Abstract A novel bacterial strain, designated KVB24T, was isolated from sea-water of Busan Harbour in South Korea. Cells of strain KVB24T were Gram-stain negative, aerobic, rod shaped and non-motile. Strain KVB24T grew optimally at 25–28 °C and pH 6.5–7.0. Based on 16S rRNA gene sequence analysis, strain KVB24T was shown to belong to the genus Lysobacter within the class Gammaproteobacteria and to be closely related to Lysobacter dokdonensis DS-58T, Lysobacter hankyongensis KTce-2T and Lysobacter niastensis GH41-7T. DNA–DNA relatedness between strain KVB24T and its current closest relative was below 70%. The predominant fatty acids of strain KVB24T were iso-C11:0, iso-C11:0 3-OH, iso-C14:0, iso-C15:0, anteiso-C15:0, iso-C16:0 and summed feature 9 comprising (iso-C17:1 ω9c and/or 10 methyl C16:0); the prominent isoprenoid was Q-8 and the major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The G + C content of genomic DNA from strain KVB24T was determined to be 67.5 mol%. Based on the phenotypic, genotypic and chemotaxonomic analyses, strain KVB24T represents a novel species of the genus Lysobacter, for which the name Lysobacter caseinilyticus sp. nov. is proposed. The type strain is KVB24T(= KACC19816T = JCM32879T).

Bacterial colonisation of reeds and cottonseed hulls in the rumen of Tarim red deer ( Cervus elaphus yarkandensis ) Abstract The rumen microbiome contributes greatly to the degradation of plant fibres to volatile fatty acids and microbial products, affecting the health and productivity of ruminants. In this study, we investigated the dynamics of colonisation by bacterial communities attached to reeds and cottonseed hulls in the rumen of Tarim red deer, a native species distributed in the desert of the Tarim Basin. The reed and cottonseed hull samples incubated in nylon bags for 1, 6, 12, and 48 h were collected and used to examine the bacterial communities by next-generation sequencing of the bacterial 16S rRNA gene. Prevotella1 and Rikenellaceae RC9 were the most abundant taxa in both the reed and cottonseed hull groups at various times, indicating a key role of these organisms in rumen fermentation in Tarim red deer. The relative abundances of cellulolytic bacteria, such as members of Fibrobacter, Treponema 2, Ruminococcaceae NK4A214 and Succiniclasticum increased, while that of the genus Prevotella 1 decreased, with increasing incubation time in both reeds and cottonseed hulls. Moreover, the temporal changes in bacterial diversity between reeds and cottonseed hulls were different, as demonstrated by the variations in the taxa Ruminococcaceae UCG 010 and Papillibacter in the reed group and Sphaerochaeta and Erysipelotrichaceae UCG 004 in the cottonseed hull group; the abundances of these bacteria first decreased and then increased. In conclusion, our results reveal the dynamics of bacterial colonisation of reeds and cottonseed hulls in the rumen of Tarim red deer.

Virulence potential of Corynebacterium striatum towards Caenorhabditis elegans Abstract Corynebacterium striatum strains have been increasingly reported as etiological agents of nosocomial infections and outbreaks in industrialized and developing countries. However, there are few studies focused on the virulence potential of C. striatum. A growing body of research supports the use of Caenorhabditis elegans as a model host for investigating the virulence potential of pathogenic bacteria, including corynebacteria. In the present study, chemotaxis behaviour, mortality, and morphological changes were investigated in nematodes infected by four C. striatum strains isolated from different clinical sites, and with different MDR profiles and PFGE types. The results showed chemotaxis of nematodes towards C. striatum. Nematode death (> 60%) was detected from the first day post-infection with all strains tested, but at different levels, independent of biofilm formation on catheter surfaces and differences in growth temperature between nematodes (20 °C) and mammals (37 °C). C. striatum 2369/II multidrug-resistant (MDR; from tracheal aspirate of a patient undergoing endotracheal intubation) and 1961/III multidrug-sensitive (MDS; urine) strains led to 100% mortality in worms. Survival of nematodes was observed until 4 days post-infection with the C. striatum 1954/IV MDS strain isolated from a surgical wound (13%) and 1987/I MDR strain isolated from a patient with a lower respiratory tract infection (39%). The Dar phenotype was observed post-infection with all MDS and MDR strains except 1954/IV. All strains showed the capacity for bagging formation. Star formation was observed only with strains that led to 100% nematode mortality. In conclusion, C. striatum was found to exert virulence for C. elegans. Variations in nematode morphological changes and levels of mortality indicate differences in the virulence potential of C. striatum independent of clinical isolation site, capacity for biofilm formation, and MDR and PFGE profiles.

Methylopila carotae sp. nov., a facultative methylotroph, isolated from a root of Daucus carota L. Abstract An aerobic facultatively methylotrophic bacterium, designated strain Das4.1T, was isolated from a root of Daucus carota L. The cells of this strain were observed to be Gram-stain negative, asporogenous, non-motile short rods multiplying by binary fission. Strain Das4.1T can utilise methanol, methylamine and a variety of polycarbon compounds as carbon and energy sources. C1-compounds were found to be assimilated via the isocitrate lyase-negative variant of the serine pathway. On medium with 0.5% methanol, growth of strain Das4.1T was observed at pH 5.5–9.0 (optimum, pH 6.0–7.0) and 18–37 °C (optimum, 24–29 °C) and in the presence of 0–2% (w/v) NaCl (optimum, 0.05%). Cells are catalase and oxidase positive and synthesise indole from l-tryptophan. The major fatty acids of methanol-grown cells were identified as C18:1ω7c, C18:0 and 11-methyl-C18:1ω7c. The predominant phospholipids were found to be phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine and phosphatidylmonomethylethanolamine. The major respiratory quinone was identified as Q-10. The DNA G + C content of strain Das4.1T was determined to be 67.3 mol% (Tm). Phylogenetic analysis based on 16S rRNA gene sequence comparison revealed that strain Das4.1T belongs to the genus Methylopila and shows high sequence similarity to Methylopila oligotropha 2395AT (98.4%) and Methylopila capsulata IM1T (98.0%). However, the DNA–DNA relatedness of strain Das4.1T with M. oligotropha 2395AT was only 22 ± 3%. Based on genotypic, chemotaxonomic and physiological characterisation, the isolate can be classified as a novel species of the genus Methylopila, for which the name Methylopila carotae sp. nov. is proposed. The type strain is Das4.1T (= VKM B-3244T = CCUG 72399T).