Publications by authors named "Manfred Rohde"

524 Publications

Filling the Gaps in the Cyanobacterial Tree of Life-Metagenome Analysis of DSM 106950, SAG 13.99 and DSM 107014.

Genes (Basel) 2021 Mar 9;12(3). Epub 2021 Mar 9.

Leibniz-Institut DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen, 38124 Braunschweig, Germany.

Cyanobacteria represent one of the most important and diverse lineages of prokaryotes with an unparalleled morphological diversity ranging from unicellular cocci and characteristic colony-formers to multicellular filamentous strains with different cell types. Sequencing of more than 1200 available reference genomes was mainly driven by their ecological relevance (, ), toxicity () and the availability of axenic strains. In the current study three slowly growing non-axenic cyanobacteria with a distant phylogenetic positioning were selected for metagenome sequencing in order to (i) investigate their genomes and to (ii) uncover the diversity of associated heterotrophs. High-throughput Illumina sequencing, metagenomic assembly and binning allowed us to establish nearly complete high-quality draft genomes of all three cyanobacteria and to determine their phylogenetic position. The cyanosphere of the limnic isolates comprises up to 40 heterotrophic bacteria that likely coexisted for several decades, and it is dominated by and . The diagnostic marker protein RpoB ensured in combination with our novel taxonomic assessment via BLASTN-dependent text-mining a reliable classification of the metagenome assembled genomes (MAGs). The detection of one new family and more than a dozen genera of uncultivated heterotrophic bacteria illustrates that non-axenic cyanobacteria are treasure troves of hidden microbial diversity.
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http://dx.doi.org/10.3390/genes12030389DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8001431PMC
March 2021

Fed-Batch - Polyhydroxyalkanoates Production in KT2440 and Δ Mutant on Biodiesel-Derived Crude Glycerol.

Front Bioeng Biotechnol 2021 16;9:642023. Epub 2021 Mar 16.

Biosystems Engineering Laboratory, Faculty of Life Sciences, Center for Bioinformatics and Integrative Biology, Universidad Andres Bello, Santiago, Chile.

Crude glycerol has emerged as a suitable feedstock for the biotechnological production of various industrial chemicals given its high surplus catalyzed by the biodiesel industry. bacteria metabolize the polyol into several biopolymers, including alginate and medium-chain-length poly(3-hydroxyalkanoates) (PHAs). Although is a suited platform to derive these polyoxoesters from crude glycerol, the attained concentrations in batch and fed-batch cultures are still low. In this study, we employed KT2440 and the hyper-PHA producer Δ mutant in two different fed-batch modes to synthesize -PHAs from raw glycerol. Initially, the cells grew in a batch phase (μ 0.21 h) for 22 h followed by a carbon-limiting exponential feeding, where the specific growth rate was set at 0.1 (h), resulting in a cell dry weight (CDW) of nearly 50 (g L) at 40 h cultivation. During the PHA production stage, we supplied the substrate at a constant rate of 50 (g h), where the KT2440 and the Δ produced 9.7 and 12.7 gPHA L, respectively, after 60 h cultivation. We next evaluated the PHA production ability of the strains using a DO-stat approach under nitrogen depletion. Citric acid was the main by-product secreted by the cells, accumulating in the culture broth up to 48 (g L) under nitrogen limitation. The mutant Δ amassed 38.9% of the CDW as -PHA and exhibited a specific PHA volumetric productivity of 0.34 (g L h), 48% higher than the parental KT2440 under the same growth conditions. The biosynthesized PHAs had average molecular weights ranging from 460 to 505 KDa and a polydispersity index (PDI) of 2.4-2.6. Here, we demonstrated that the DO-stat feeding approach in high cell density cultures enables the high yield production of -PHA in strains using the industrial crude glycerol, where the fed-batch process selection is essential to exploit the superior biopolymer production hallmarks of engineered bacterial strains.
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http://dx.doi.org/10.3389/fbioe.2021.642023DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8007980PMC
March 2021

Loss of Hem1 disrupts macrophage function and impacts migration, phagocytosis, and integrin-mediated adhesion.

Curr Biol 2021 Mar 6. Epub 2021 Mar 6.

Department of Cell Biology, Helmholtz Centre for Infection Research (HZI), Braunschweig, Germany. Electronic address:

Hematopoietic-specific protein 1 (Hem1) is an essential subunit of the WAVE regulatory complex (WRC) in immune cells. WRC is crucial for Arp2/3 complex activation and the protrusion of branched actin filament networks. Moreover, Hem1 loss of function in immune cells causes autoimmune diseases in humans. Here, we show that genetic removal of Hem1 in macrophages diminishes frequency and efficacy of phagocytosis as well as phagocytic cup formation in addition to defects in lamellipodial protrusion and migration. Moreover, Hem1-null macrophages displayed strong defects in cell adhesion despite unaltered podosome formation and concomitant extracellular matrix degradation. Specifically, dynamics of both adhesion and de-adhesion as well as concomitant phosphorylation of paxillin and focal adhesion kinase (FAK) were significantly compromised. Accordingly, disruption of WRC function in non-hematopoietic cells coincided with both defects in adhesion turnover and altered FAK and paxillin phosphorylation. Consistently, platelets exhibited reduced adhesion and diminished integrin αIIbβ3 activation upon WRC removal. Interestingly, adhesion phenotypes, but not lamellipodia formation, were partially rescued by small molecule activation of FAK. A full rescue of the phenotype, including lamellipodia formation, required not only the presence of WRCs but also their binding to and activation by Rac. Collectively, our results uncover that WRC impacts on integrin-dependent processes in a FAK-dependent manner, controlling formation and dismantling of adhesions, relevant for properly grabbing onto extracellular surfaces and particles during cell edge expansion, like in migration or phagocytosis.
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http://dx.doi.org/10.1016/j.cub.2021.02.043DOI Listing
March 2021

The Two-Component System 09 Regulates Pneumococcal Carbohydrate Metabolism and Capsule Expression.

Microorganisms 2021 Feb 24;9(3). Epub 2021 Feb 24.

Department of Molecular Genetics and Infection Biology, Interfaculty Institute for Genetics and Functional Genomics, Center for Functional Genomics of Microbes, University of Greifswald, 17487 Greifswald, Germany.

two-component regulatory systems (TCSs) are important systems that perceive and respond to various host environmental stimuli. In this study, we have explored the role of TCS09 on gene expression and phenotypic alterations in D39. Our comparative transcriptomic analyses identified 67 differently expressed genes in total. Among those, and the operon involved in galactose metabolism showed the highest changes. Intriguingly, the encapsulated and nonencapsulated -mutants showed significant growth defects under nutrient-defined conditions, in particular with galactose as a carbon source. Phenotypic analyses revealed alterations in the morphology of the nonencapsulated - and -mutants, whereas the encapsulated - and -mutants produced higher amounts of capsule. Interestingly, the encapsulated D39∆ showed only the opaque colony morphology, while the D39∆- and D39∆-mutants had a higher proportion of transparent variants. The phenotypic variations of D39ΔΔ and D39ΔΔ are in accordance with their higher numbers of outer membrane vesicles, higher sensitivity against Triton X-100 induced autolysis, and lower resistance against oxidative stress. In conclusion, these results indicate the importance of TCS09 for pneumococcal metabolic fitness and resistance against oxidative stress by regulating the carbohydrate metabolism and thereby, most likely indirectly, the cell wall integrity and amount of capsular polysaccharide.
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http://dx.doi.org/10.3390/microorganisms9030468DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7996280PMC
February 2021

Streptomyces bathyalis sp. nov., an actinobacterium isolated from the sponge in a deep sea.

Antonie Van Leeuwenhoek 2021 Apr 17;114(4):425-435. Epub 2021 Feb 17.

Microbial Strain Collection (MISG), Helmholtz Centre for Infection Research (HZI), 38124, Braunschweig, Germany.

A novel actinobacterium, designated ASO4wet, was isolated from the unidentified sponge (SO4) in the deep sea collected of the North Atlantic Ocean. Study of 16S rRNA gene sequences indicated that strain ASO4wet is a member of the genus Streptomyces and showed the closest similarities to Streptomyces karpasiensis K413 (98.87 %), Streptomyces glycovorans YIM M 10366 (98.38 %), and Streptomyces abyssalis YIM M 10400 (97.53 %). Strain ASO4wet contained MK-9(H8) as the predominant menaquinone and the major fatty acids are iso-C, anteiso-C, and iso-C. Polyphasic taxonomy was carried out between strain ASO4wet and its phylogenetically closely related Streptomyces strains, which further elucidated their relatedness and revealed that strain ASO4wet could be distinguished from currently known Streptomyces species. Strain ASO4wet clearly represents a novel species in genus Streptomyces. We propose the name Streptomyces bathyalis sp. nov., with the type strain ASO4wet (= DSM 106605 = NCCB 100657). Analysis of the whole-genome sequence of S. bathyalis revealed that genome size is 7,377,472 bp with 6332 coding sequences.
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http://dx.doi.org/10.1007/s10482-021-01528-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7979654PMC
April 2021

Dinoroseobacter shibae Outer Membrane Vesicles Are Enriched for the Chromosome Dimer Resolution Site .

mSystems 2021 Jan 12;6(1). Epub 2021 Jan 12.

Institute of Microbiology, Technical University of Braunschweig, Braunschweig, Germany

Outer membrane vesicles (OMVs) are universally produced by prokaryotes and play important roles in symbiotic and pathogenic interactions. They often contain DNA, but a mechanism for its incorporation is lacking. Here, we show that , a dinoflagellate symbiont, constitutively secretes OMVs containing DNA. Time-lapse microscopy captured instances of multiple OMV production at the septum during cell division. DNA from the vesicle lumen was up to 22-fold enriched for the region around the terminus of replication (). The peak of coverage was located at a conserved 28-bp palindromic sequence required for binding of the site-specific tyrosine recombinases XerC/XerD. These enzymes are activated at the last stage of cell division immediately prior to septum formation when they are bound by the divisome protein FtsK. We suggest that overreplicated regions around the terminus have been repaired by the FtsK-XerC/XerD molecular machinery. The vesicle proteome was clearly dominated by outer membrane and periplasmic proteins. Some of the most abundant vesicle membrane proteins were predicted to be required for direct interaction with peptidoglycan during cell division (LysM, Tol-Pal, Spol, lytic murein transglycosylase). OMVs were 15-fold enriched for the saturated fatty acid 16:00. We hypothesize that constitutive OMV secretion in is coupled to cell division. The footprint of the FtsK--XerC/XerD molecular machinery suggests a novel potentially highly conserved route for incorporation of DNA into OMVs. Clearing the division site from small DNA fragments might be an important function of vesicles produced during exponential growth under optimal conditions. Gram-negative bacteria continually form vesicles from their outer membrane (outer membrane vesicles [OMVs]) during normal growth. OMVs frequently contain DNA, and it is unclear how DNA can be shuffled from the cytoplasm to the OMVs. We studied OMV cargo in , a symbiont of dinoflagellates, using microscopy and a multi-omics approach. We found that vesicles formed during undisturbed exponential growth contain DNA which is enriched for genes around the replication terminus, specifically, the binding site for an enzyme complex that is activated at the last stage of cell division. We suggest that the enriched genes are the result of overreplication which is repaired by their excision and excretion via membrane vesicles to clear the divisome from waste DNA.
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http://dx.doi.org/10.1128/mSystems.00693-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7901474PMC
January 2021

sp. nov., a new member of the Actinomycetes from a hot desert in Iran.

Int J Syst Evol Microbiol 2021 Feb;71(2)

Microbial Strain Collection, Helmholtz Centre for Infection Research, Inhoffenstrasse 7, D-38124 Braunschweig, Germany.

Isolate 4NS15 was isolated from a neglected arid habitat in Kerman, Iran. The strain showed 16S rRNA gene sequence similarity values of 98.9 % to the type strains of subsp. , and and 98.6 % to the type strain subsp. , respectively. Genome-based phylogenetic analysis revealed that isolate 4NS15 is closely related to subsp. DSM 43828. The digital DNA-DNA hybridization value between the genome sequences of 4NS15 and strain DSM 43828 is 29.8 %. Strain 4NS15 produces long chains of spores without a sporangium-like structure which can be distinguished from other species. Isolate 4NS15 has a genome size of 10.35 Mbp with a G+C content of 68.1 mol%. Whole-cell hydrolysates of isolate 4NS15 are rich in -diaminopimelic acid and cell-wall sugars such as arabinose, galactose, glucose and ribose. Major fatty acids (>10 %) are C, iso-C and iso-C. The phospholipid profile contains diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylhydroxyethanolamine, aminolipid and glycoaminolipid. The predominant menaquinone is MK-9(H). Based on its phenotypic and genotypic characteristics, isolate 4NS15 (NCCB 100701=CIP 111705=DSM 110728) merits recognition as representing a novel species of the genus , for which the name sp. nov. is proposed.
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http://dx.doi.org/10.1099/ijsem.0.004625DOI Listing
February 2021

Cultivation-Independent Analysis of the Bacterial Community Associated With the Calcareous Sponge and Isolation of Gen. Nov., Sp. Nov., Belonging to the Barely Studied Class in the Phylum .

Front Microbiol 2020 22;11:602250. Epub 2020 Dec 22.

Department of Microbiology, Radboud University, Nijmegen, Netherlands.

Marine ecosystems serve as global carbon sinks and nutrient source or breeding ground for aquatic animals. Sponges are ancient parts of these important ecosystems and can be found in caves, the deep-sea, clear waters, or more turbid environments. Here, we studied the bacterial community composition of the calcareous sponge sampled close to the island Corsica in the Mediterranean Sea with an emphasis on planctomycetes. We show that the phylum accounts for 9% of the -associated bacterial community, a 5-fold enrichment compared to the surrounding seawater. Indeed, the use of as a yet untapped source of novel planctomycetal strains led to the isolation of strain KS4. The strain represents a novel genus and species within the class in the phylum and displays interesting cell biological features, such as formation of outer membrane vesicles and an unexpected mode of cell division.
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http://dx.doi.org/10.3389/fmicb.2020.602250DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7783415PMC
December 2020

Bordetella bronchiseptica promotes adherence, colonization, and cytotoxicity of Streptococcus suis in a porcine precision-cut lung slice model.

Virulence 2021 12;12(1):84-95

Institute for Microbiology, University of Veterinary Medicine Hannover , Hannover, Germany.

and are major pathogens in pigs, which are frequently isolated from co-infections in the respiratory tract and contribute to the porcine respiratory disease complex (PRDC). Despite the high impact of co-infections on respiratory diseases of swine (and other hosts), very little is known about pathogen-pathogen-host interactions and the mechanisms of pathogenesis. In the present study, we established a porcine precision-cut lung slice (PCLS) model to analyze the effects of infection on adherence, colonization, and cytotoxic effects of . We hypothesized that induction of ciliostasis by a clinical isolate of may promote subsequent infection with a virulent serotype 2 strain. To investigate this theory, we monitored the ciliary activity by light microscopy, measured the release of lactate dehydrogenase, and calculated the number of PCLS-associated bacteria. To study the role of the pore-forming toxin suilysin (SLY) in -induced cytotoxicity, we included a SLY-negative isogenic mutant and the complemented mutant strain. Furthermore, we analyzed infected PCLS by histopathology, immunofluorescence microscopy, and field emission scanning electron microscopy. Our results showed that pre-infection with promoted adherence, colonization, and, as a consequence of the increased colonization, the cytotoxic effects of , probably by reduction of the ciliary activity. Moreover, cytotoxicity induced by is strictly dependent on the presence of SLY. Though the underlying molecular mechanisms remain to be fully clarified, our results clearly support the hypothesis that paves the way for infection.
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http://dx.doi.org/10.1080/21505594.2020.1858604DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7781633PMC
December 2021

In Vitro Evaluation of a Phage Cocktail Controlling Infections with .

Viruses 2020 12 19;12(12). Epub 2020 Dec 19.

Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, Inhoffenstraße 7B, 38124 Braunschweig, Germany.

Worldwide, poultry industry suffers from infections caused by avian pathogenic . Therapeutic failure due to resistant bacteria is of increasing concern and poses a threat to human and animal health. This causes a high demand to find alternatives to fight bacterial infections in animal farming. Bacteriophages are being especially considered for the control of multi-drug resistant bacteria due to their high specificity and lack of serious side effects. Therefore, the study aimed on characterizing phages and composing a phage cocktail suitable for the prevention of infections with . Six phages were isolated or selected from our collections and characterized individually and in combination with regard to host range, stability, reproduction, and efficacy in vitro. The cocktail consisting of six phages was able to inhibit formation of biofilms by some strains but not by all. Phage-resistant variants arose when bacterial cells were challenged with a single phage but not when challenged by a combination of four or six phages. Resistant variants arising showed changes in carbon metabolism and/or motility. Genomic comparison of wild type and phage-resistant mutant E28.G28R3 revealed a deletion of several genes putatively involved in phage adsorption and infection.
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http://dx.doi.org/10.3390/v12121470DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7768485PMC
December 2020

Pneumolysin induces platelet destruction, not platelet activation, which can be prevented by immunoglobulin preparations in vitro.

Blood Adv 2020 12;4(24):6315-6326

Department of Transfusion Medicine, Institute of Immunology and Transfusion Medicine, University Medicine Greifswald, Greifswald, Germany.

Community-acquired pneumonia by primary or superinfections with Streptococcus pneumoniae can lead to acute respiratory distress requiring mechanical ventilation. The pore-forming toxin pneumolysin alters the alveolar-capillary barrier and causes extravasation of protein-rich fluid into the interstitial pulmonary tissue, which impairs gas exchange. Platelets usually prevent endothelial leakage in inflamed pulmonary tissue by sealing inflammation-induced endothelial gaps. We not only confirm that S pneumoniae induces CD62P expression in platelets, but we also show that, in the presence of pneumolysin, CD62P expression is not associated with platelet activation. Pneumolysin induces pores in the platelet membrane, which allow anti-CD62P antibodies to stain the intracellular CD62P without platelet activation. Pneumolysin treatment also results in calcium efflux, increase in light transmission by platelet lysis (not aggregation), loss of platelet thrombus formation in the flow chamber, and loss of pore-sealing capacity of platelets in the Boyden chamber. Specific anti-pneumolysin monoclonal and polyclonal antibodies inhibit these effects of pneumolysin on platelets as do polyvalent human immunoglobulins. In a post hoc analysis of the prospective randomized phase 2 CIGMA trial, we show that administration of a polyvalent immunoglobulin preparation was associated with a nominally higher platelet count and nominally improved survival in patients with severe S pneumoniae-related community-acquired pneumonia. Although, due to the low number of patients, no definitive conclusion can be made, our findings provide a rationale for investigation of pharmacologic immunoglobulin preparations to target pneumolysin by polyvalent immunoglobulin preparations in severe community-acquired pneumococcal pneumonia, to counteract the risk of these patients becoming ventilation dependent. This trial was registered at www.clinicaltrials.gov as #NCT01420744.
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http://dx.doi.org/10.1182/bloodadvances.2020002372DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7756997PMC
December 2020

Analysis of bacterial communities in a municipal duck pond during a phytoplankton bloom and isolation of Anatilimnocola aggregata gen. nov., sp. nov., Lacipirellula limnantheis sp. nov. and Urbifossiella limnaea gen. nov., sp. nov. belonging to the phylum Planctomycetes.

Environ Microbiol 2021 Mar 12;23(3):1379-1396. Epub 2021 Jan 12.

Department of Microbial Interactions, Institute of Microbiology, Friedrich Schiller University, Jena, Germany.

Waterbodies such as lakes and ponds are fragile environments affected by human influences. Suitable conditions can result in massive growth of phototrophs, commonly referred to as phytoplankton blooms. Such events benefit heterotrophic bacteria able to use compounds secreted by phototrophs or their biomass as major nutrient source. One example of such bacteria are Planctomycetes, which are abundant on the surfaces of marine macroscopic phototrophs; however, less data are available on their ecological roles in limnic environments. In this study, we followed a cultivation-independent deep sequencing approach to study the bacterial community composition during a cyanobacterial bloom event in a municipal duck pond. In addition to cyanobacteria, which caused the bloom event, members of the phylum Planctomycetes were significantly enriched in the cyanobacteria-attached fraction compared to the free-living fraction. Separate datasets based on isolated DNA and RNA point towards considerable differences in the abundance and activity of planctomycetal families, indicating different activity peaks of these families during the cyanobacterial bloom. Motivated by the finding that the sampling location harbours untapped bacterial diversity, we included a complementary cultivation-dependent approach and isolated and characterized three novel limnic strains belonging to the phylum Planctomycetes.
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http://dx.doi.org/10.1111/1462-2920.15341DOI Listing
March 2021

Additions to the genus Gimesia: description of Gimesia alba sp. nov., Gimesia algae sp. nov., Gimesia aquarii sp. nov., Gimesia aquatilis sp. nov., Gimesia fumaroli sp. nov. and Gimesia panareensis sp. nov., isolated from aquatic habitats of the Northern Hemisphere.

Antonie Van Leeuwenhoek 2020 Dec 24;113(12):1999-2018. Epub 2020 Nov 24.

Department of Microbial Interactions, Friedrich-Schiller-University, Jena, Germany.

Thirteen novel planctomycetal strains were isolated from five different aquatic sampling locations. These comprise the hydrothermal vent system close to Panarea Island (Italy), a biofilm on the surface of kelp at Monterey Bay (CA, USA), sediment and algae on Mallorca Island (Spain) and Helgoland Island (Germany), as well as a seawater aquarium in Braunschweig, Germany. All strains were shown to belong to the genus Gimesia. Their genomes cover a size range from 7.22 to 8.29 Mb and have a G+C content between 45.1 and 53.7%. All strains are mesophilic (T 26-33 °C) with generation times between 12 and 32 h. Analysis of fatty acids yielded palmitic acid (16:0) and a fatty acid with the equivalent chain length of 15.817 as major compounds. While five of the novel strains belong to the already described species Gimesia maris and Gimesia chilikensis, the other strains belong to novel species, for which we propose the names Gimesia alba (type strain Pan241w = DSM 100744 = LMG 31345 = CECT 9841 = VKM B-3430), Gimesia algae (type strain Pan161 = CECT 30192 = STH00943 = LMG 29130), Gimesia aquarii (type strain V144 = DSM 101710 = VKM B-3433), Gimesia fumaroli (type strain Enr17 = DSM 100710 = VKM B-3429) and Gimesia panareensis (type strain Enr10 = DSM 100416 = LMG 29082). STH numbers refer to the Jena Microbial Resource Collection (JMRC).
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http://dx.doi.org/10.1007/s10482-020-01489-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7716864PMC
December 2020

Unsaturated Fatty Acids Control Biofilm Formation of and Other Gram-Positive Bacteria.

Antibiotics (Basel) 2020 Nov 8;9(11). Epub 2020 Nov 8.

Chemical Microbiology, Helmholtz Centre for Infection Research (HZI), Inhoffenstraße 7, 38124 Braunschweig, Germany.

Infections involving biofilms are difficult to treat due to increased resistances against antibiotics and the immune system. Hence, there is an urgent demand for novel drugs against biofilm infections. During our search for novel biofilm inhibitors from fungi, we isolated linoleic acid from the ascomycete which showed biofilm inhibition of several bacteria at sub-MIC concentrations. Many fatty acids possess antimicrobial activities, but their minimum inhibitory concentrations (MIC) are high and reports on biofilm interferences are scarce. We demonstrated that not only linoleic acid but several unsaturated long-chain fatty acids inhibited biofilms at sub-MIC concentrations. The antibiofilm activity exerted by long-chain fatty acids was mainly against Gram-positive bacteria, especially against . Micrographs of treated biofilms revealed a reduction in the extracellular polymeric substances, pointing to a possible mode of action of fatty acids on biofilms. The fatty acids had a strong species specificity. Poly-unsaturated fatty acids had higher activities than saturated ones, but no obvious rule could be found for the optimal length and desaturation for maximal activity. As free fatty acids are non-toxic and ubiquitous in food, they may offer a novel tool, especially in combination with antibiotics, for the control of biofilm infections.
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http://dx.doi.org/10.3390/antibiotics9110788DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7695168PMC
November 2020

Updates to the recently introduced family Lacipirellulaceae in the phylum Planctomycetes: isolation of strains belonging to the novel genera Aeoliella, Botrimarina, Pirellulimonas and Pseudobythopirellula and the novel species Bythopirellula polymerisocia and Posidoniimonas corsicana.

Antonie Van Leeuwenhoek 2020 Dec 5;113(12):1979-1997. Epub 2020 Nov 5.

Department of Microbiology, Radboud Universiteit, Nijmegen, The Netherlands.

Eight novel strains of the phylum Planctomycetes were isolated from different aquatic habitats. Among these habitats were the hydrothermal vent system close to Panarea Island, a public beach at Mallorca Island, the shore of Costa Brava (Spain), and three sites with brackish water in the Baltic Sea. The genome sizes of the novel strains range from 4.33 to 6.29 Mb with DNA G+C contents between 52.8 and 66.7%. All strains are mesophilic (T 24-30 °C) and display generation times between 17 and 94 h. All eight isolates constitute novel species of either already described or novel genera within the family Lacipirellulaceae. Two of the novel species, Posidoniimonas polymericola (type strain Pla123a = DSM 103020 = LMG 29466) and Bythopirellula polymerisocia (type strain Pla144 = DSM 104841 = VKM B-3442), belong to established genera, while the other strains represent the novel genera Aeoliella gen. nov., Botrimarina gen. nov., Pirellulimonas gen. nov. and Pseudobythopirellula gen. nov. Based on our polyphasic analysis, we propose the species Aeoliella mucimassa sp. nov. (type strain Pan181 = DSM 29370 = LMG 31346 = CECT 9840 = VKM B-3426), Botrimarina colliarenosi sp. nov. (type strain Pla108 = DSM 103355 = LMG 29803), Botrimarina hoheduenensis sp. nov. (type strain Pla111 = DSM 103485 = STH00945, Jena Microbial Resource Collection JMRC), Botrimarina mediterranea sp. nov. (type strain Spa11 = DSM 100745 = LMG 31350 = CECT 9852 = VKM B-3431), Pirellulimonas nuda sp. nov. (type strain Pla175 = DSM 109594 = CECT 9871 = VKM B-3448) and Pseudobythopirellula maris sp. nov. (type strain Mal64 = DSM 100832 = LMG 29020).
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http://dx.doi.org/10.1007/s10482-020-01486-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7717034PMC
December 2020

Host-induced spermidine production in motile triggers phagocytic uptake.

Elife 2020 09 22;9. Epub 2020 Sep 22.

Department of Molecular Bacteriology, Helmholtz Centre for Infection Research, Braunschweig, Germany.

Exploring the complexity of host-pathogen communication is vital to understand why microbes persist within a host, while others are cleared. Here, we employed a dual-sequencing approach to unravel conversational turn-taking of dynamic host-pathogen communications. We demonstrate that upon hitting a host cell, motile induce a specific gene expression program. This results in the expression of spermidine on the surface, which specifically activates the PIP-pathway to induce phagocytic uptake into primary or immortalized murine cells. Non-motile bacteria are more immunogenic due to a lower expression of upon host-cell contact, but do not produce spermidine and are phagocytosed less. We demonstrate that not only the presence of pathogen inherent molecular patterns induces immune responses, but that bacterial motility is linked to a host-cell-induced expression of additional immune modulators. Our results emphasize on the value of integrating microbiological and immunological findings to unravel complex and dynamic host-pathogen interactions.
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http://dx.doi.org/10.7554/eLife.55744DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7538158PMC
September 2020

Author Correction: The planctomycete Stieleria maiorica Mal15 employs stieleriacines to alter the species composition in marine biofilms.

Commun Biol 2020 Sep 21;3(1):531. Epub 2020 Sep 21.

Department of Microbiology, Radboud University, Nijmegen, The Netherlands.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.
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http://dx.doi.org/10.1038/s42003-020-01271-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7506533PMC
September 2020

Toll-like Receptor 5 Activation by the CagY Repeat Domains of Helicobacter pylori.

Cell Rep 2020 09;32(11):108159

Department of Biology, Division of Microbiology, Friedrich Alexander University Erlangen-Nuremberg, 91058 Erlangen, Germany. Electronic address:

Helicobacter pylori (Hp) is an important human pathogen associated with gastric inflammation and neoplasia. It is commonly believed that this bacterium avoids major immune recognition by Toll-like receptors (TLRs) because of low intrinsic activity of its flagellin and lipopolysaccharides (LPS). In particular, TLR5 specifically detects flagellins in various bacterial pathogens, while Hp evolved mutations in flagellin to evade detection through TLR5. Cancerogenic Hp strains encode a type IV secretion system (T4SS). The T4SS core component and pilus-associated protein CagY, a large VirB10 ortholog, drives effector molecule translocation. Here, we identify CagY as a flagellin-independent TLR5 agonist. We detect five TLR5 interaction sites, promoting binding of CagY-positive Hp to TLR5-expressing cells, TLR5 stimulation, and intracellular signal transduction. Consequently, CagY constitutes a remarkable VirB10 member detected by TLR5, driving crucial innate immune responses by this human pathogen.
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http://dx.doi.org/10.1016/j.celrep.2020.108159DOI Listing
September 2020

Author Correction: The planctomycete Stieleria maiorica Mal15 employs stieleriacines to alter the species composition in marine biofilms.

Commun Biol 2020 Aug 31;3(1):487. Epub 2020 Aug 31.

Department of Microbiology, Radboud University, Nijmegen, The Netherlands.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.
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http://dx.doi.org/10.1038/s42003-020-01228-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7458906PMC
August 2020

Three Planctomycetes isolated from biotic surfaces in the Mediterranean Sea and the Pacific Ocean constitute the novel species Symmachiella dynata gen. nov., sp. nov. and Symmachiella macrocystis sp. nov.

Antonie Van Leeuwenhoek 2020 Dec 24;113(12):1965-1977. Epub 2020 Aug 24.

Department of Microbial Interactions, Friedrich-Schiller-University, Jena, Germany.

Planctomycetes is a phylum of environmentally important bacteria, which also receive significant attention due to their fascinating cell biology. Access to axenic Planctomycete cultures is crucial to study cell biological features within this phylum in further detail. In this study, we characterise three novel strains, Mal52, Pan258 and CA54, which were isolated close to the coasts of the islands Mallorca (Spain) and Panarea (Italy), and from Monterey Bay, CA, USA. The three isolates show optimal growth at temperatures between 22 and 24 °C and at pH 7.5, divide by polar budding, lack pigmentation and form strong aggregates in liquid culture. Analysis of five phylogenetic markers suggests that the strains constitute two novel species within a novel genus in the family Planctomycetaceae. The strains Mal52 (DSM 101177 = VKM B-3432) and Pan258 were assigned to the species Symmachiella dynata gen nov., sp. nov., while strain CA54 (DSM 104301 = VKM B-3450) forms a separate species of the same genus, for which we propose the name Symmachiella macrocystis sp. nov.
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http://dx.doi.org/10.1007/s10482-020-01464-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7716862PMC
December 2020

Stieleria varia sp. nov., isolated from wood particles in the Baltic Sea, constitutes a novel species in the family Pirellulaceae within the phylum Planctomycetes.

Antonie Van Leeuwenhoek 2020 Dec 14;113(12):1953-1963. Epub 2020 Aug 14.

Department of Microbiology, Radboud University, Nijmegen, The Netherlands.

Species belonging to the bacterial phylum Planctomycetes are ubiquitous members of the microbial communities in aquatic environments and are frequently isolated from various biotic and abiotic surfaces in marine and limnic water bodies. Planctomycetes have large genomes of up to 12.4 Mb, follow complex lifestyles and display an uncommon cell biology; features which motivate the investigation of members of this phylum in greater detail. As a contribution to the current collection of axenic cultures of Planctomycetes, we here describe strain Pla52 isolated from wood particles in the Baltic Sea. Phylogenetic analysis places the strain in the family Pirellulaceae and suggests two species of the recently described genus Stieleria as current closest neighbours. Strain Pla52n shows typical features of members of the class Planctomycetia, including division by polar budding and the presence of crateriform structures. Colonies of strain Pla52n have a light orange colour, which is an unusual pigmentation compared to the majority of members in the phylum, which show either a pink to red pigmentation or entirely lack pigmentation. Optimal growth of strain Pla52n at 33 °C and pH 7.5 indicates a mesophilic (i.e. with optimal growth between 20 and 45 °C) and neutrophilic growth profile. The strain is an aerobic heterotroph with motile daughter cells. Its genome has a size of 9.6 Mb and a G + C content of 56.0%. Polyphasic analyses justify delineation of the strain from described species within the genus Stieleria. Therefore, we conclude that strain Pla52n = LMG 29463 = VKM B-3447 should be classified as the type strain of a novel species, for which we propose the name Stieleria varia sp. nov.
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http://dx.doi.org/10.1007/s10482-020-01456-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7717043PMC
December 2020

Non-Typeable Invade Choroid Plexus Epithelial Cells in a Polar Fashion.

Int J Mol Sci 2020 Aug 10;21(16). Epub 2020 Aug 10.

Pediatric Infectious Diseases, Department of Pediatrics, Medical Faculty Mannheim, Heidelberg University, Theodor-Kutzer-Ufer 1-3, D-68167 Mannheim, Germany.

Non-typeable (NTHI) is a pathogen of the human respiratory tract causing the majority of invasive infections. Severe invasive infections such as septicemia and meningitis occur rarely, but the lack of a protecting vaccine and the increasing antibiotic resistance of NTHI impede treatment and emphasize its relevance as a potential meningitis causing pathogen. Meningitis results from pathogens crossing blood-brain barriers and invading the immune privileged central nervous system (CNS). In this study, we addressed the potential of NTHI to enter the brain by invading cells of the choroid plexus (CP) prior to meningeal inflammation to enlighten NTHI pathophysiological mechanisms. A cell culture model of human CP epithelial cells, which form the blood-cerebrospinal fluid barrier (BCSFB) in vivo, was used to analyze adhesion and invasion by immunofluorescence and electron microscopy. NTHI invade CP cells in vitro in a polar fashion from the blood-facing side. Furthermore, NTHI invasion rates are increased compared to encapsulated HiB and HiF strains. Fimbriae occurrence attenuated adhesion and invasion. Thus, our findings underline the role of the BCSFB as a potential entry port for NTHI into the brain and provide strong evidence for a function of the CP during NTHI invasion into the CNS during the course of meningitis.
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http://dx.doi.org/10.3390/ijms21165739DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7461124PMC
August 2020

Stieleriacines, -Acyl Dehydrotyrosines From the Marine Planctomycete sp. nov.

Front Microbiol 2020 16;11:1408. Epub 2020 Jul 16.

Department of Microbial Drugs, Helmholtz Centre for Infection Research (HZI), Braunschweig, Germany.

Bacteria of the phylum occur ubiquitously in marine environments and play important roles in the marine nitrogen- and carbon cycle, for example as scavengers after phototrophic blooms. Here, we describe the isolation and characterization of the planctomycetal strain Enr13 isolated from a sp. biofilm obtained from seawater sediment close to Panarea Island, Italy. Phylogenetic tree reconstruction based on 16S rRNA gene sequences and multi-locus sequence analysis supports the delineation of strain Enr13 from characterized species part of the phylum of . HPLC-MS analysis of culture broth obtained from strain Enr13 revealed the presence of lipophilic metabolites, of which the major compound was isolated by preparative reversed-phase HPLC. The structure of this compound, named stieleriacine D (), was elucidated utilizing HRESIMS, 1D- and 2D-NMR data as a new -acylated dehydrotyrosine derivative. Its biosynthesis was proposed based on an gene cluster analysis. Through analysis of the MS/MS spectrum of and its minor derivative, stieleriacine E (), it was possible to assign the structure of without isolation. showed antibacterial activity, however, the wide distribution of structurally related compounds indicates a potential role as a signaling molecule.
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http://dx.doi.org/10.3389/fmicb.2020.01408DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7378531PMC
July 2020

gen. nov., sp. nov., a novel prosthecate, budding member of the family isolated from forest soil.

Int J Syst Evol Microbiol 2020 Sep 7;70(9):4966-4977. Epub 2020 Aug 7.

Braunschweig University of Technology, Spielmanstraße 7, 38106 Braunschweig, Germany.

The family comprises prosthecate bacteria with a dimorphic cell cycle and also non-prosthecate bacteria. Cells of all described species divide by binary fission. Strain 0127_4 was isolated from forest soil in Baden Württemberg (Germany) and determined to be the first representative of the family which divided by budding. Cells of strain 0127_4 were Gram-negative, rod-shaped, prosthecate, motile by means of a polar flagellum, non-spore-forming and non-capsulated. The strain formed small white colonies and grew aerobically and chemo-organotrophically utilizing organic acids, amino acids and proteinaceous substrates. 16S rRNA gene sequence analysis indicated that this bacterium was related to TH1-2 and DRW22-8 with 91.3 and 89.7% sequence similarity, respectively. Four unidentified glycolipids were detected as the major polar lipids and, unlike all described members of the family , phosphatidylglycerol was absent. The major fatty acids were summed feature 8 (Cω7/Cω6), summed feature 9 (iso-Cω9/C 10-methyl), C and summed feature 3 (C6/C7). The major respiratory quinone was Q-10. The G+C content of the genomic DNA was 63.5 %. Based on the present taxonomic characterization, strain 0127_4 represents a novel species of a new genus, gen. nov., sp. nov. The type strain of is 0127_4 (=DSM 104635=CECT 9243).
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http://dx.doi.org/10.1099/ijsem.0.004367DOI Listing
September 2020

Symbiosis between nanohaloarchaeon and haloarchaeon is based on utilization of different polysaccharides.

Proc Natl Acad Sci U S A 2020 08 5;117(33):20223-20234. Epub 2020 Aug 5.

Institute for Biological Resources and Marine Biotechnologies, Italian National Research Council, 98122 Messina, Italy;

Nano-sized archaeota, with their small genomes and limited metabolic capabilities, are known to associate with other microbes, thereby compensating for their own auxotrophies. These diminutive and yet ubiquitous organisms thrive in hypersaline habitats that they share with haloarchaea. Here, we reveal the genetic and physiological nature of a nanohaloarchaeon-haloarchaeon association, with both microbes obtained from a solar saltern and reproducibly cultivated together in vitro. The nanohaloarchaeon Nanohalobium constans LC1Nh is an aerotolerant, sugar-fermenting anaerobe, lacking key anabolic machinery and respiratory complexes. The nanohaloarchaeon cells are found physically connected to the chitinolytic haloarchaeon sp. LC1Hm. Our experiments revealed that this haloarchaeon can hydrolyze chitin outside the cell (to produce the monosaccharide -acetylglucosamine), using this beta-glucan to obtain carbon and energy for growth. However, LC1Hm could not metabolize either glycogen or starch (both alpha-glucans) or other polysaccharides tested. Remarkably, the nanohaloarchaeon's ability to hydrolyze glycogen and starch to glucose enabled growth of sp. LC1Hm in the absence of a chitin. These findings indicated that the nanohaloarchaeon-haloarchaeon association is both mutualistic and symbiotic; in this case, each microbe relies on its partner's ability to degrade different polysaccharides. This suggests, in turn, that other nano-sized archaeota may also be beneficial for their hosts. Given that availability of carbon substrates can vary both spatially and temporarily, the susceptibility of to colonization by Nanohalobium can be interpreted as a strategy to maximize the long-term fitness of the host.
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http://dx.doi.org/10.1073/pnas.2007232117DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7443923PMC
August 2020

The Microbiome of Seagrass Leaves Can Be Dominated by Planctomycetes.

Front Microbiol 2020 10;11:1458. Epub 2020 Jul 10.

Department of Microbiology, Radboud University, Nijmegen, Netherlands.

Seagrass meadows are ubiquitous, fragile and endangered marine habitats, which serve as fish breeding grounds, stabilize ocean floor substrates, retain nutrients and serve as important carbon sinks, counteracting climate change. In the Mediterranean Sea, seagrass meadows are mostly formed by the slow-growing endemic plant (Neptune grass), which is endangered by global warming and recreational motorboating. Despite its importance, surprisingly little is known about the leaf surface microbiome of . Using amplicon sequencing, we here show that species belonging to the phylum can dominate the biofilms of young and aged leaves. Application of selective cultivation techniques allowed for the isolation of two novel planctomycetal strains belonging to two yet uncharacterized genera.
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http://dx.doi.org/10.3389/fmicb.2020.01458DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7366357PMC
July 2020

Rosistilla oblonga gen. nov., sp. nov. and Rosistilla carotiformis sp. nov., isolated from biotic or abiotic surfaces in Northern Germany, Mallorca, Spain and California, USA.

Antonie Van Leeuwenhoek 2020 Dec 4;113(12):1939-1952. Epub 2020 Jul 4.

Department of Microbial Interactions, Friedrich Schiller University, Jena, Germany.

Planctomycetes are ubiquitous bacteria with fascinating cell biological features. Strains available as axenic cultures in most cases have been isolated from aquatic environments and serve as a basis to study planctomycetal cell biology and interactions in further detail. As a contribution to the current collection of axenic cultures, here we characterise three closely related strains, Poly24, CA51 and Mal33, which were isolated from the Baltic Sea, the Pacific Ocean and the Mediterranean Sea, respectively. The strains display cell biological features typical for related Planctomycetes, such as division by polar budding, presence of crateriform structures and formation of rosettes. Optimal growth was observed at temperatures of 30-33 °C and at pH 7.5, which led to maximal growth rates of 0.065-0.079 h, corresponding to generation times of 9-11 h. The genomes of the novel isolates have a size of 7.3-7.5 Mb and a G + C content of 57.7-58.2%. Phylogenetic analyses place the strains in the family Pirellulaceae and suggest that Roseimaritima ulvae and Roseimaritima sediminicola are the current closest relatives. Analysis of five different phylogenetic markers, however, supports the delineation of the strains from members of the genus Roseimaritima and other characterised genera in the family. Supported by morphological and physiological differences, we conclude that the strains belong to the novel genus Rosistilla gen. nov. and constitute two novel species, for which we propose the names Rosistilla carotiformis sp. nov. and Rosistilla oblonga sp. nov. (the type species). The two novel species are represented by the type strains Poly24 (= DSM 102938 = VKM B-3434 = LMG 31347 = CECT 9848) and CA51 (= DSM 104080 = LMG 29702), respectively.
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http://dx.doi.org/10.1007/s10482-020-01441-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7716947PMC
December 2020

Maioricimonas rarisocia gen. nov., sp. nov., a novel planctomycete isolated from marine sediments close to Mallorca Island.

Antonie Van Leeuwenhoek 2020 Dec 25;113(12):1901-1913. Epub 2020 Jun 25.

Department of Microbiology, Radboud Universiteit, Nijmegen, The Netherlands.

Planctomycetes are ubiquitous bacteria with environmental and biotechnological relevance. Axenic cultures of planctomycetal strains are the basis to analyse their unusual biology and largely uncharacterised metabolism in more detail. Here, we describe strain Mal4 isolated from marine sediments close to Palma de Mallorca, Spain. Strain Mal4 displays common planctomycetal features, such as division by polar budding and the presence of fimbriae and crateriform structures on the cell surface. Cell growth was observed at ranges of 10-39 °C (optimum at 31 °C) and pH 6.5-9.0 (optimum at 7.5). The novel strain shows as pear-shaped cells of 2.0 ± 0.2 × 1.4 ± 0.1 µm and is one of the rare examples of orange colony-forming Planctomycetes. Its genome has a size of 7.7 Mb with a G+C content of 63.4%. Phylogenetically, we conclude that strain Mal4 (= DSM 100296= LMG 29133) is the type strain representing the type species of a novel genus, for which we propose the name Maioricimonas rarisocia gen. nov., sp. nov.
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http://dx.doi.org/10.1007/s10482-020-01436-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7716917PMC
December 2020

Thalassoglobus polymorphus sp. nov., a novel Planctomycete isolated close to a public beach of Mallorca Island.

Antonie Van Leeuwenhoek 2020 Dec 24;113(12):1915-1926. Epub 2020 Jun 24.

Department of Microbiology, Radboud Universiteit, Nijmegen, The Netherlands.

Access to axenic cultures is crucial to extend the knowledge of the biology, lifestyle or metabolic capabilities of bacteria from different phyla. The phylum Planctomycetes is an excellent example since its members display an unusual cell biology and complex lifestyles. As a contribution to the current collection of axenic planctomycete cultures, here we describe strain Mal48 isolated from phytoplankton material sampled at the coast of S'Arenal close to Palma de Mallorca (Spain). The isolated strain shows optimal growth at pH 7.0-7.5 and 30 °C and exhibits typical features of Planctomycetes. Cells of the strain are spherical to pear-shaped, divide by polar budding with daughter cells showing the same shape as the mother cell, tend to aggregate, display a stalk and produce matrix or fimbriae. Strain Mal48 showed 95.8% 16S rRNA gene sequence similarity with the recently described Thalassoglobus neptunius KOR42. The genome sequence of the novel isolate has a size of 6,357,355 bp with a G+C content of 50.3%. A total of 4874 protein-coding genes, 41 tRNA genes and 2 copies of the 16S rRNA gene are encoded in the genome. Based on phylogenetic, morphological and physiological analyses, we conclude that strain Mal48 (= DSM 100737 = LMG 29019) should be classified as the type strain of a new species in the genus Thalassoglobus, for which the name Thalassoglobus polymorphus sp. nov. is proposed.
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http://dx.doi.org/10.1007/s10482-020-01437-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7716918PMC
December 2020

Caulifigura coniformis gen. nov., sp. nov., a novel member of the family Planctomycetaceae isolated from a red biofilm sampled in a hydrothermal area.

Antonie Van Leeuwenhoek 2020 Dec 24;113(12):1927-1937. Epub 2020 Jun 24.

Department of Microbial Interactions, Institute of Microbiology, Friedrich Schiller University, Jena, Germany.

Pan44, a novel strain belonging to the phylum Planctomycetes, was isolated from a red biofilm in a hydrothermal area close to the island Panarea in the Tyrrhenian Sea north of Sicily, Italy. The strain forms white colonies on solid medium and displays the following characteristics: cell division by budding, formation of rosettes, presence of matrix or fimbriae and long stalks. The cell surface has an interesting and characteristic texture made up of triangles and rectangles, which leads to a pine cone-like morphology of the strain. Strain Pan44 is mesophilic (temperature optimum 26 °C), slightly alkaliphilic (pH optimum 8.0), aerobic and heterotrophic. The strain has a genome size of 6.76 Mb with a G + C content of 63.2%. Phylogenetically, the strain is a member of the family Planctomycetaceae, order Planctomycetales, class Planctomycetia. Our analysis supports delineation of strain Pan44 from all known genera in this family, hence, we propose to assign it to a novel species within a novel genus, for which we propose the name Caulifigura coniformis gen. nov., sp. nov., represented by Pan44 (DSM 29405 = LMG 29788) as the type strain.
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http://dx.doi.org/10.1007/s10482-020-01439-wDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7717036PMC
December 2020