Neotype Strain Research Articles

Complete genome sequence of the methanogenic neotype strain Methanobacterium formicicum MFT

The neotype strain Methanobacterium formicicum MFT (DSM1535), a hydrogenotrophic methanogenic Archaeon, was isolated from a domestic sewage sludge digestor in Urbana (IL, USA). Here, the complete genome sequence of the methanogen is reported. The genome is 2,478,074bp in size, featuring a GC content of 41.23%. M. formicicum MFT encodes several genes predicted to be involved in adaptation to abiotic stress such as high osmolarity. The strain MFT is of biotechnological importance since M. formicicum strains are often found in production-scale biogas plants and it is suggested as a starter culture for the anaerobic biomethanation process.

Designation of type strains for seven species of the order Myxococcales and proposal for neotype strains of Cystobacter ferrugineus, Cystobacter minus and Polyangium fumosum

Ten species of the order Myxococcales with validly published names are devoid of living type strains. Four species of the genus Chondromyces are represented by dead herbarium samples as the type material. For a species of the genus Melittangium and two species of the genus Polyangium, no physical type material was assigned at the time of validation of the names or later on. In accordance with rule 18f of the International Code of Nomenclature of Bacteria the following type strains are designated for these species: strain Cm a14(T) ( = DSM 14605(T) = JCM 12615(T)) as the type strain of Chondromyces apiculatus, strain Cm c5(T) ( = DSM 14714(T) = JCM 12616(T)) as the type strain of Chondromyces crocatus, strain Sy t2(T) ( = DSM 14631(T) = JCM 12617(T)) as the type strain of Chondromyces lanuginosus, strain Cm p51(T) ( = DSM 14607(T) = JCM 12618(T)) as the type strain of Chondromyces pediculatus, strain Me b8(T) ( = DSM 14713(T) = JCM 12633(T)) as the type strain of Melittangium boletus, strain Pl s12(T) ( = DSM 14670(T) = JCM 12637(T)) as the type strain of Polyangium sorediatum and strain Pl sm5(T) ( = DSM 14734(T) = JCM 12638(T)) as the type strain of Polyangium spumosum. Furthermore, the type strains given for three species of the genera Cystobacter and Polyangium had been kept at one university institute and have been lost according to our investigations. In accordance with Rule 18c of the Bacteriological Code, we propose the following neotype strains: strain Cb fe18 ( = DSM 14716 = JCM 12624) as the neotype strain of Cystobacter ferrugineus, strain Cb m2 ( = DSM 14751 = JCM 12627) as the neotype strain of Cystobacter minus and strain Pl fu5 ( = DSM 14668 = JCM 12636) as the neotype strain of Polyangium fumosum. The proposals of the strains are based on the descriptions and strain proposals given in the respective chapters of Bergey's Manual of Systematic Bacteriology (2005).

Mycocin sensitivity patterns of Kluyveromyces species: Kluyveromyces sensu lato vs. Kluyveromyces sensu stricto

The Kluyveromyces species reassigned to the genera Lachancea and Vanderwaltozyma are insensitive to five mycocins secreted by Pichia membranifaciens. The remaining Kluyveromyces species including species transferred to the genera Kazachstania, Nakaseomyces, and Tetrapisispora are sensitive to them. Only the neotype strain is insensitive to mycocins among Kluyveromyces lactis cultures.

Genome sequence of Wickerhamomyces anomalus DSM 6766 reveals genetic basis of biotechnologically important antimicrobial activities

The ascomycetous yeast Wickerhamomyces anomalus (formerly Pichia anomala and Hansenula anomala) exhibits antimicrobial activities and flavoring features that are responsible for its frequent association with food, beverage and feed products. However, limited information on the genetic background of this yeast and its multiple capabilities are currently available. Here, we present the draft genome sequence of the neotype strain W.anomalus DSM 6766. On the basis of pyrosequencing, a de novo assembly of this strain resulted in a draft genome sequence with a total size of 25.47Mbp. An automatic annotation using RAPYD generated 11512 protein-coding sequences. This annotation provided the basis to analyse metabolic capabilities, phylogenetic relationships, as well as biotechnologically important features and yielded novel candidate genes of W.anomalus DSM 6766 coding for proteins participating in antimicrobial activities.

Genome sequence of the filamentous, gliding Thiothrix nivea neotype strain (JP2(T)).

Thiothrix nivea (Rabenhorst 1865) Winogradsky 1888 (Approved Lists 1980) emend. Larkin and Shinabarger 1983 is the type species of the genus Thiothrix in the family Thiotrichaceae. The species is of interest not only because of its isolated location in the yet to be genomically characterized region of the tree of life, but also because of its life-style with gliding gonidia, the multilayer sheath, rosettes, and the embedded sulfur granules. Strain JP2T is the neotype strain of the species which was first observed by Rabenhorst in 1865 and later reclassified by Winogradsky in 1888 into the then novel genus Thiothrix. This is the first completed (improved-high-quality-draft) genome sequence to be published of a member of the family Thiotrichaceae. The genome in its current assembly consists of 15 contigs in four scaffolds with a total of 4,691,711 bp bearing 4,542 protein-coding and 52 RNA genes and is a part of the Genomic Encyclopedia of Bacteria and Archaea project.

Genetic diversity and mycotoxin production of Fusarium lactis species complex isolates from sweet pepper

An internal fruit rot disease of sweet peppers was first detected in Belgium in 2003. Research conducted mostly in Canada indicates that this disease is primarily caused by Fusarium lactis Pirotta. Ninety-eight Fusarium isolates obtained from diseased sweet peppers from Belgium, as well as from other countries (Canada, the Netherlands and the United Kingdom) were identified by sequencing the translation elongation factor 1α (EF). Of these 98 isolates, 13 were identified as F. oxysporum Schltdl., nine as F. proliferatum (Matsush.) Nirenberg and two belonged to clade 3 of the F. solani species complex. Of the 74 remaining isolates, the EF sequence showed 97% to 98% similarity to F. lactis. Of these isolates, the β-tubulin (TUB), calmodulin (CAM) and the second largest subunit of RNA polymerase II (RPB2) genes were also sequenced. Analysis of the combined sequences revealed that the 74 isolates share nine combined sequences that correspond to nine multilocus sequence types (STs), while the F. lactis neotype strain and one other strain, both isolated from figs, form a separate ST. Together, these 10 STs represent a monophyletic F. lactis species complex (FLASC). An unusually high level of genetic diversity was observed between (groups of) these STs. Two of them (ST5 and ST6) fulfilled the criteria for species recognition based on genealogical exclusivity and together represent a new monophyletic species lineage (FLASC-1). The seven other STs, together with the F. lactis neotype ST, form a paraphyletic species lineage in the African clade of the Gibberella fujikuroi species complex (GFSC). From each of the 10 STs, the mycotoxin production was assessed using a multi-mycotoxin liquid chromatography mass spectrometry method. Out of the 27 analyzed mycotoxins, beauvericin and fumonisins were detected in sweet pepper tissue and in maize kernels. The 10 STs clearly differed in the amount of mycotoxin produced, but there was only limited congruence between the production profile and the phylogenetic analysis. Furthermore, the morphological characterization (based on mycelial growth rate and the length of macroconidia) showed distinct differences between the 10 STs, but again there was limited congruence with the phylogenetic results. In conclusion, the data presented in this study demonstrate that 75% of the isolates obtained from sweet pepper with internal fruit rot belong to a F. lactis species complex (FLASC), including a new FLASC-1 monophyletic species, and that the members of this complex display great genetic and phenotypic diversity.

The status of the species Beijerinckia fluminensis Döbereiner and Ruschel 1958. Request for an Opinion

In a previous article [Oggerin M., Arahal, D. R., Rubio, V. & Marin, I. (2009). Int J Syst Evol Microbiol 59, 2323-2328], it has been shown that strain Beijerinckia fluminensis UQM 1685(T) and its derived equivalent B. fluminensis CIP 106281(T) do not conform to the description of the type strain of Beijerinckia fluminensis Döbereiner and Ruschel 1958. Indeed, both strains were identified as members of the species Rhizobium radiobacter and exhibited marked phenotypic and genotypic differences with members of the genus Beijerinckia. It was concluded that both strains, and any other equivalents derived from them, do not descend from the nomenclatural type. Since then, our attempts to find older deposits of the type strain, hopefully derived from the original isolate, or other existing strains of Beijerinckia fluminensis that could be proposed as a neotype strain, have been in vain. It is therefore proposed that the Judicial Commission should place the name Beijerinckia fluminensis Döbereiner and Ruschel 1958 on the list of rejected names if a suitable replacement type strain or a neotype cannot be found within two years following the publication of this Request (Rule 18c).

An azole-resistant isolate of Malassezia pachydermatis

Canine Malassezia dermatitis (MD) is frequently treated with systemic ketoconazole (KTZ) and itaconazole (ITZ). However, the antifungal susceptibility of clinical isolates of M. pachydermatis from dogs and cats to the azoles has not been well investigated. In the present study, the in vitro susceptibility of the standard strain (CBS1879: the neotype strain of M. pachydermatis) and 29 clinical isolates of M. pachydermatis to the azoles was measured by a modified CLSI M27-A2 test using modified Dixon medium as well as by the E-test. The minimum inhibitory concentrations (MICs) of the 30 isolates of M. pachydermatis (including the neotype strain) against KTZ and ITZ were <0.03μg/ml by the two methods. The MICs of 1 clinical isolate (ASC-11) were 1 and 2μg/ml against KTZ, and 2 and 8μg/ml against ITZ, by the modified CLSI M27-A2 test and the E-test, respectively. Thus, isolate ASC-11 may be resistant to these azoles, making this the first report of a resistant isolate of M. pachydermatis to KTZ and ITZ.

Study of Glycerol and Sweet Water as a Carbon Source for Production of Rhamnolipids by Naturally Occurring Strains of Pseudomonas aeruginosa (ATCC 10145 and ATCC 9027)

Abstract The interest in biosurfactant has increased considerably due to their large potential for industrial applications. Rhamnolipid is a simplest class of biosurfactants with well defined structure. The main obstacle in commercialization of biosurfactant especially rhamnolipids is the higher cost of the production. Glycerol is readily available from commercial fat-splitting process in the form of sweet water (up to 20% w/v) at very low cost. Two natural strains of Pseudomonas aeruginosa (ATCC 10145 and ATCC 9027) were compared for rhamnolipid production. The neotype strain of Pseudomonas aeruginosa (ATCC 10145) was found to produce more amounts of rhamnolipids than Pseudomonas aeruginosa (ATCC 9027) on glucose and glycerol as carbon source. The microbial growth (0.94 g/L) and formation of rhamnolipids (2.75 g/L) was maximum at 5% (w/v) glycerol concentration by Pseudomonas aeruginosa (ATCC 10145). The fermentation was monitored for 8 days on shake flask and magnetically stirred batch culture. In magnetically stirred batch culture, microbial growth was 0.69 g/L and 2.73 g/L of rhamnolipid was formed. Sweet water was also used as a source of glycerol.

An Asymmetric Model for Na+-translocating Glutaconyl-CoA Decarboxylases

Glutaconyl-CoA decarboxylase (Gcd) couples the biotin-dependent decarboxylation of glutaconyl-CoA with the generation of an electrochemical Na(+) gradient. Sequencing of the genes encoding all subunits of the Clostridium symbiosum decarboxylase membrane complex revealed that it comprises two distinct biotin carrier subunits, GcdC(1) and GcdC(2), which differ in the length of a central alanine- and proline-rich linker domain. Co-crystallization of the decarboxylase subunit GcdA with the substrate glutaconyl-CoA, the product crotonyl-CoA, and the substrate analogue glutaryl-CoA, respectively, resulted in a high resolution model for substrate binding and catalysis revealing remarkable structural changes upon substrate binding. Unlike the GcdA structure from Acidaminococcus fermentans, these data suggest that in intact Gcd complexes, GcdA is associated as a tetramer crisscrossed by a network of solvent-filled tunnels.

Emended descriptions of the genera Myxococcus and Corallococcus, typification of the species Myxococcus stipitatus and Myxococcus macrosporus and a proposal that they be represented by neotype strains. Request for an Opinion

The genus Corallococcus was separated from the genus Myxococcus mainly on the basis of differences in morphology and consistency of swarms and fruiting bodies of the respective members. Phylogenetic, chemotaxonomic and physiological evidence is presented here that underpins the separate status of these phylogenetically neighbouring genera. Emended descriptions of the two genera are presented. The data also suggest that the species Corallococcus macrosporus belongs to the genus Myxococcus. To the best of our knowledge, the type strains of the species Myxococcus macrosporus and Myxococcus stipitatus are not available from any established culture collection or any other source. A Request for an Opinion is made regarding the proposal that strain Cc m8 (=DSM 14697=CIP 109128) be formally recognized as the neotype strain for the species Myxococcus macrosporus, replacing the designated type strain Windsor M271T, and that strain Mx s8 (=DSM 14675=JCM 12634) be formally recognized as the neotype strain for the species Myxococcus stipitatus, replacing the designated type strain Windsor M78T.

Malassezia furfur fingerprints as possible markers for human phylogeography

Malassezia furfur was the first species described within the cosmopolitan yeast genus Malassezia, which now comprises 13 species. Reported isolation rates of these species from healthy and diseased human skin show geographic variations. PCR-fingerprinting with the wild-type phage M13 primer (5'-GAGGGTGGCGGTTCT-3') was applied to investigate phylogeographic associations of M. furfur strains isolated from Scandinavians residing permanently in Greece, in comparison to clinical isolates from Greek, Bulgarian and Chinese native residents. Seven M. furfur strains from Scandinavians were compared with the Neotype strain (CBS1878), CBS global collection strains (n=10) and clinical isolates from Greece (n=4), Bulgaria (n=15) and China (n=6). Scandinavian, Greek and Bulgarian M. furfur strains mostly formed distinct group clusters, providing initial evidence for an association with the host's geographical origin and with the underlying skin condition. These initial data address the hypothesis that M. furfur could be a eukaryotic candidate eligible for phylogeographic studies.

Proposal of a neotype strain (A1-86) for Eubacterium rectale. Request for an Opinion

Eubacterium rectale is one of the most abundant bacterial species recovered from human faeces. E. rectale (Hauduroy et al. 1937) appears in the 'List of Bacterial Names with Standing in Nomenclature', but it is noted that the originally proposed type strain, VPI 0989(T), has been lost and its possible replacement by another strain (VPI 0990) from the same faecal sample has never been formally proposed. It is therefore proposed that strain A1-86 (=DSM 17629=NCIMB 14373), isolated from human adult faeces, be formally recognized as the neotype strain of Eubacterium rectale.

Analysis of the Genome Sequence ofLactobacillus gasseriATCC 33323 Reveals the Molecular Basis of an Autochthonous Intestinal Organism

This study presents the complete genome sequence of Lactobacillus gasseri ATCC 33323, a neotype strain of human origin and a native species found commonly in the gastrointestinal tracts of neonates and adults. The plasmid-free genome was 1,894,360 bp in size and predicted to encode 1,810 genes. The GC content was 35.3%, similar to the GC content of its closest relatives, L. johnsonii NCC 533 (34%) and L. acidophilus NCFM (34%). Two identical copies of the prophage LgaI (40,086 bp), of the Sfi11-like Siphoviridae phage family, were integrated tandomly in the chromosome. A number of unique features were identified in the genome of L. gasseri that were likely acquired by horizontal gene transfer and may contribute to the survival of this bacterium in its ecological niche. L. gasseri encodes two restriction and modification systems, which may limit bacteriophage infection. L. gasseri also encodes an operon for production of heteropolysaccharides of high complexity. A unique alternative sigma factor was present similar to that of B. caccae ATCC 43185, a bacterial species isolated from human feces. In addition, L. gasseri encoded the highest number of putative mucus-binding proteins (14) among lactobacilli sequenced to date. Selected phenotypic characteristics that were compared between ATCC 33323 and other human L. gasseri strains included carbohydrate fermentation patterns, growth and survival in bile, oxalate degradation, and adhesion to intestinal epithelial cells, in vitro. The results from this study indicated high intraspecies variability from a genome encoding traits important for survival and retention in the gastrointestinal tract.

Identification of dermatophytes by sequence analysis of the rRNA gene internal transcribed spacer regions

Identification of dermatophytes using the traditional method is sometimes problematic because of atypical microscopic or macroscopic morphology. The aim of this study was to evaluate the feasibility of using sequencing of the ribosomal internal transcribed spacer (ITS)1 and ITS2 regions for identification of 17 dermatophyte species. The ITS regions of 188 strains (62 reference strains and 126 clinical isolates) were amplified by PCR and sequenced. Species identification was made by sequence comparison with an in-house database comprising ITS sequences of type or neotype strains or by blast searches for homologous sequences in public databases. Strains producing discrepant results between conventional methods and ITS sequence analysis were analysed further by sequencing the D1-D2 domain of the large-subunit rRNA gene for species clarification. The identification rates by ITS1 and ITS2 sequencing were higher than 97 %. Based on reference sequences of type or neotype strains, it was noted that most strains of Trichophyton mentagrophytes were misidentifications of Trichophyton interdigitale. In addition, barcode sequences were present in species of the Microsporum canis complex and Trichophyton rubrum complex. These barcode sequences are useful for species delineation when the results of ITS sequencing are ambiguous. In conclusion, ITS sequencing provides a very accurate and useful method for the identification of dermatophytes.

Genetic typing of Malassezia pachydermatis from different domestic animals

In the present study, random amplification of polymorphic DNA, which detects DNA polymorphism in fungal genomic DNA, was applied for genetic typing of Malassezia pachydermatis isolates. Fifty-five isolates from different domestic animals and body sites and the neotype strain CBS 1879 were characterized. Primers M13 and OPT-20 were used to analyse their genetic relatedness and similarity. This technique allowed us to distinguish four different genetic types. The predominant genetic type was observed in isolates recovered from different anatomical locations in all animals. It was the only genetic type found in cats, horse, goat and pig. The other three genetic types were observed only in isolates from external ear canals of dogs. Types II and IV were only recovered from external otitic ears and type III from healthy ears. An animal was colonised by more than one type of M. pachydermatis and different genetic types were detected in the same body site. Some genetic types were only isolated from diseased skin.

Ribotyping ofLactobacillus casei group strains isolated from dairy products

A series of lactobacilli isolated from dairy products were characterized using biotyping and ribotyping with EcoRI and HindIII restriction enzymes. Biotyping assigned 14 strains as Lactobacillus casei, 6 strains as Lactobacillus paracasei subsp. paracasei and 12 as Lactobacillus rhamnosus. The obtained ribotype patterns separated all analyzed strains into two clearly distinguished groups corresponding to L. rhamnosus and L. casei/L. paracasei subsp. paracasei. The HindIII ribotypes of individual strains representing these two groups were visually very similar. In contrast, EcoRI ribotyping revealed high intraspecies variability. All ribotypes of L. casei and L. paracasei subsp. paracasei dairy strains were very close and some strains even shared identical ribotype profiles. The type strains L. casei CCM 7088T (= ATCC 393T) and Lactobacillus zeae CCM 7069T revealing similar ribopatterns formed a separate subcluster using both restriction enzymes. In contrast, the ribotype profile of L. casei CCM 7089 (= ATCC 334) was very close to ribopatterns obtained from the dairy strains. These results support synonymy of L. casei and L. paracasei species revealed by other studies as well as reclassification of the type strain L. casei CCM 7088T as L. zeae and designation of L. casei CCM 7089 as the neotype strain.

Morphological investigation of three Tetramitus spp. which are phylogenetically very closely related: Tetramitus horticolus, Tetramitus russelli n. comb. and Tetramitus pararusselli n. sp.

Two vahlkampfiid amoebae have been isolated which have the same 5.8S rDNA sequence as Tetramitus horticolus, but which differ in internal transcribed spacer (ITS) sequences from this species. The two new strains, 123R and Rhodos, are identical with each other in ITS sequences, except for a 1 bp substitution in the ITS1 and an insertion–deletion sequence (indel) of 4 bp in the ITS2. Because of the close phylogenetic relationship between the two new isolates and T. horticolus we compared both isolates with the latter species by light and electron microscopy. While T. horticolus has a cyst pore structure not seen in any other vahlkampfiid, the two new strains have no pores in their cysts. Strains 123R and Rhodos are very similar in morphology, but significantly different in cyst diameter. Neither strain could be induced to form flagellates under laboratory conditions. Strain 123R seems to correspond to the description of Vahlkampfia russelli. Because of this, and its molecular identification as a strain of Tetramitus, we propose that this strain should become the neotype strain of V. russelli (Singh, 1952), now classified as T. russelli n. comb. Because of the small (but significant) differences of strain Rhodos from strain 123R we propose the former as the type strain of a new species, T. pararusselli n. sp. The addition of T. horticolus, T. russelli and T. pararusselli confirms that the phenotypic definition of the genus Tetramitus is obsolete. Instead the genus can be differentiated by the significant differences from other vahlkampfiid genera in rDNA sequence.

Isolation of a vahlkampfiid amoeba from a contact lens: Tetramitus ovis (Schmidt, 1913) n. comb.

A vahlkampfiid amoeba has been isolated from a contact lens. Based on 5.8S rDNA sequence analysis the strain belongs to the genus Tetramitus. The internal transcribed spacer (ITS1 and ITS2) sequences differ by 3 and 9 bp, respectively, from T. lobospinosus. The morphology of the cyst does not correspond to T. lobospinosus, but is identical to that published for Vahlkampfia ovis at the beginning of the last century. There is no reference strain of V. ovis to investigate using molecular techniques. Therefore, we propose that the strain under study should be considered to represent the neotype strain of V. ovis, now classified as T. ovis (Schmidt, 1913) n. comb.

Rejection of the genus name Pelczaria with the species Pelczaria aurantia Poston 1994. Opinion 78

The Judicial Commission of the International Committee for Systematics of Prokaryotes has decided to place the genus Pelczaria with the species Pelczaria aurantia on the list of nomina rejicienda, due to the lack of an authentic type or neotype strain.