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  1. Article ; Online: Phenotypic and genotypic discrimination of Francisella tularensis ssp. holarctica clades

    Kristin Köppen / Kerstin Rydzewski / Joerg Doellinger / Kerstin Myrtennäs / Mats Forsman / Sandra Appelt / Holger Scholz / Klaus Heuner

    International Journal of Medical Microbiology, Vol 313, Iss 4, Pp 151583- (2023)

    2023  

    Abstract: Francisella tularensis is the causative agent of tularemia, a zoonotic disease with a wide host range. F. tularensis ssp. holarctica (Fth) is of clinical relevance for European countries, including Germany. Whole genome sequencing methods, including ... ...

    Abstract Francisella tularensis is the causative agent of tularemia, a zoonotic disease with a wide host range. F. tularensis ssp. holarctica (Fth) is of clinical relevance for European countries, including Germany. Whole genome sequencing methods, including canonical Single Nucleotide Polymorphism (canSNP) typing and whole genome SNP typing, have revealed that European Fth strains belong to a few monophyletic populations. The majority of German Fth isolates belong to two basal phylogenetic clades B.6 (biovar I) and B.12 (biovar II). Strains of B.6 and B.12 seem to differ in their pathogenicity, and it has been shown that strains of biovar II are resistant against erythromycin. In this study, we present data corroborating our previous data demonstrating that basal clade B.12 can be divided into clades B.71 and B.72. By applying phylogenetic whole genome analysis as well as proteome analysis, we could verify that strains of these two clades are distinct from one another. This was confirmed by measuring the intensity of backscatter light on bacteria grown in liquid media. Strains belonging to clades B.6, B.71 or B.72 showed clade-specific backscatter growth curves. Furthermore, we present the whole genome sequence of strain A-1341, as a reference genome of clade B.71, and whole proteomes comparison of Fth strains belonging to clades B.6, B.71 and B.72. Further research is necessary to investigate phenotypes and putative differences in pathogenicity of the investigated different clades of Fth to better understand the relationship between observed phenotypes, pathogenicity and distribution of Fth strains.
    Keywords Francisella ; Phylogenetic clade B.71 ; B.12 ; B.6 ; Proteomics ; Mass spectrometry ; Microbiology ; QR1-502 ; Other systems of medicine ; RZ201-999
    Subject code 572
    Language English
    Publishing date 2023-07-01T00:00:00Z
    Publisher Elsevier
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  2. Article ; Online: Francisella tularensis Subspecies holarctica and Tularemia in Germany

    Sandra Appelt / Mirko Faber / Kristin Köppen / Daniela Jacob / Roland Grunow / Klaus Heuner

    Microorganisms, Vol 8, Iss 1448, p

    2020  Volume 1448

    Abstract: Tularemia is a zoonotic disease caused by Francisella tularensis a small, pleomorphic, facultative intracellular bacterium. In Europe, infections in animals and humans are caused mainly by Francisella tularensis subspecies holarctica . Humans can be ... ...

    Abstract Tularemia is a zoonotic disease caused by Francisella tularensis a small, pleomorphic, facultative intracellular bacterium. In Europe, infections in animals and humans are caused mainly by Francisella tularensis subspecies holarctica . Humans can be exposed to the pathogen directly and indirectly through contact with sick animals, carcasses, mosquitoes and ticks, environmental sources such as contaminated water or soil, and food. So far, F. tularensis subsp. holarctica is the only Francisella species known to cause tularemia in Germany. On the basis of surveillance data, outbreak investigations, and literature, we review herein the epidemiological situation—noteworthy clinical cases next to genetic diversity of F. tularensis subsp. holarctica strains isolated from patients. In the last 15 years, the yearly number of notified cases of tularemia has increased steadily in Germany, suggesting that the disease is re-emerging. By sequencing F. tularensis subsp. holarctica genomes, knowledge has been added to recent findings, completing the picture of genotypic diversity and geographical segregation of Francisella clades in Germany. Here, we also shortly summarize the current knowledge about a new Francisella species ( Francisella sp. strain W12-1067) that has been recently identified in Germany. This species is the second Francisella species discovered in Germany.
    Keywords Francisella tularensis subspecies holarctica ; W12-1067 ; tularemia ; rabbit fever ; zoonotic disease ; Germany ; Biology (General) ; QH301-705.5
    Subject code 590
    Language English
    Publishing date 2020-09-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  3. Article ; Online: Metabolic adaption of Legionella pneumophila during intracellular growth in Acanthamoeba castellanii

    Mareike Kunze / Thomas Steiner / Fan Chen / Claudia Huber / Kerstin Rydzewski / Maren Stämmler / Klaus Heuner / Wolfgang Eisenreich

    International Journal of Medical Microbiology, Vol 311, Iss 4, Pp 151504- (2021)

    2021  

    Abstract: The metabolism of Legionella pneumophila strain Paris was elucidated during different time intervals of growth within its natural host Acanthamoeba castellanii. For this purpose, the amoebae were supplied after bacterial infection (t =0 h) with 11 mM [U- ... ...

    Abstract The metabolism of Legionella pneumophila strain Paris was elucidated during different time intervals of growth within its natural host Acanthamoeba castellanii. For this purpose, the amoebae were supplied after bacterial infection (t =0 h) with 11 mM [U-13C6]glucose or 3 mM [U-13C3]serine, respectively, during 0−17 h, 17−25 h, or 25−27 h of incubation. At the end of these time intervals, bacterial and amoebal fractions were separated. Each of these fractions was hydrolyzed under acidic conditions. 13C-Enrichments and isotopologue distributions of resulting amino acids and 3-hydroxybutyrate were determined by gas chromatography – mass spectrometry. Comparative analysis of the labelling patterns revealed the substrate preferences, metabolic pathways, and relative carbon fluxes of the intracellular bacteria and their amoebal host during the time course of the infection cycle. Generally, the bacterial infection increased the usage of exogenous glucose via glycolysis by A. castellanii. In contrast, carbon fluxes via the amoebal citrate cycle were not affected. During the whole infection cycle, intracellular L. pneumophila incorporated amino acids from their host into the bacterial proteins. However, partial bacterial de novo biosynthesis from exogenous 13C-Ser and, at minor rates, from 13C-glucose could be shown for bacterial Ala, Asp, Glu, and Gly. More specifically, the catabolic usage of Ser increased during the post-exponential phase of intracellular growth, whereas glucose was utilized by the bacteria throughout the infection cycle and not only late during infection as assumed on the basis of earlier in vitro experiments. The early usage of 13C-glucose by the intracellular bacteria suggests that glucose availability could serve as a trigger for replication of L. pneumophila inside the vacuoles of host cells.
    Keywords Legionella pneumophila ; Acanthamoeba castellanii ; Patho-metabolism ; Bipartite metabolism ; Stable isotope labelling ; Host‐pathogen interaction ; Microbiology ; QR1-502 ; Other systems of medicine ; RZ201-999
    Subject code 572
    Language English
    Publishing date 2021-05-01T00:00:00Z
    Publisher Elsevier
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  4. Article ; Online: Myo-Inositol as a carbon substrate in Francisella and insights into the metabolism of Francisella sp. strain W12-1067

    Fan Chen / Kristin Köppen / Kerstin Rydzewski / Rosa Einenkel / Clara Morguet / Duc Tung Vu / Wolfgang Eisenreich / Klaus Heuner

    International Journal of Medical Microbiology, Vol 310, Iss 4, Pp 151426- (2020)

    2020  

    Abstract: Recently, a new environmental Francisella strain, Francisella sp. strain W12-1067, has been identified in Germany. This strain is negative for the Francisella pathogenicity island (FPI) but exhibits a putative alternative type VI secretion system. Some ... ...

    Abstract Recently, a new environmental Francisella strain, Francisella sp. strain W12-1067, has been identified in Germany. This strain is negative for the Francisella pathogenicity island (FPI) but exhibits a putative alternative type VI secretion system. Some known virulence factors of Francisella are present, but the pathogenic capacity of this species is not known yet. In silico genome analysis reveals the presence of a gene cluster tentatively enabling myo-inositol (MI) utilization via a putative inositol oxygenase. Labelling experiments starting from 2H-inositol demonstrate that this gene cluster is indeed involved in the metabolism of MI. We further show that, under in vitro conditions, supply of MI increases growth rates of strain W12-1067 in the absence of glucose and that the metabolism of MI is strongly reduced in a W12-1067 mutant lacking the MI gene cluster. The positive growth effect of MI in the absence of glucose is restored in this mutant strain by introducing the complete MI gene cluster. F. novicida Fx1 is also positive for the MI metabolizing gene cluster and MI again increases growth in a glucose-free medium, in contrast to F. novicida strain U112, which is shown to be a natural mutant of the MI metabolizing gene cluster. Labelling experiments of Francisella sp. strain W12-1067 in medium T containing 13C-glucose, 13C-serine or 13C-glycerol as tracers suggest a bipartite metabolism where glucose is mainly metabolized through glycolysis, but not through the Entner-Doudoroff pathway or the pentose phosphate pathway. Carbon flux from 13C-glycerol and 13C-serine is less active, and label from these tracers is transferred mostly into amino acids, lactate and fatty acids. Together, the metabolism of Francisella sp. strain W12-1067 seems to be more related to the respective one in F. novicida rather than in F. tularensis subsp. holarctica.
    Keywords Francisella sp. W12-1067 ; Metabolism ; Glucose ; Myo-inositol ; Isotopolog profiling ; Allofrancisella ; Microbiology ; QR1-502 ; Other systems of medicine ; RZ201-999
    Subject code 570
    Language English
    Publishing date 2020-05-01T00:00:00Z
    Publisher Elsevier
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  5. Article: Successful re-evaluation of broth medium T for growth of Francisella tularensis ssp. and other highly pathogenic bacteria

    Becker, Silke / Daniela Jacob / Klaus Heuner / Petra Lochau / Roland Grunow

    Journal of microbiological methods. 2016 Feb., v. 121

    2016  

    Abstract: Pavlovich's medium T was compared with other broadly used media and extensively checked by growth of various subspecies of Francisellatularensis as well as other risk group 3 bacteria. The medium was successfully re-evaluated as an optimal liquid medium ... ...

    Abstract Pavlovich's medium T was compared with other broadly used media and extensively checked by growth of various subspecies of Francisellatularensis as well as other risk group 3 bacteria. The medium was successfully re-evaluated as an optimal liquid medium suitable for enrichment of fastidious and/or highly pathogenic bacteria.
    Keywords bacteria ; Francisella tularensis ; risk
    Language English
    Dates of publication 2016-02
    Size p. 5-7.
    Publishing place Elsevier B.V.
    Document type Article
    ZDB-ID 604916-3
    ISSN 1872-8359 ; 0167-7012
    ISSN (online) 1872-8359
    ISSN 0167-7012
    DOI 10.1016/j.mimet.2015.11.018
    Database NAL-Catalogue (AGRICOLA)

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  6. Article ; Online: Outbreak of Tularemia in a Group of Hunters in Germany in 2018—Kinetics of Antibody and Cytokine Responses

    Daniela Jacob / Anne Barduhn / Dennis Tappe / Jessica Rauch / Klaus Heuner / Daniela Hierhammer / Katharina vom Berge / Julia M. Riehm / Matthias Hanczaruk / Stefanie Böhm / Merle M. Böhmer / Regina Konrad / Berit Bouschery / Marc Dauer / Elisabeth Schichtl / Hamid Hossain / Roland Grunow

    Microorganisms, Vol 8, Iss 1645, p

    2020  Volume 1645

    Abstract: In November 2018, an outbreak of tularemia occurred among hare hunters in Bavaria, Germany. At least one infected hare was confirmed as the source of infection. A number of hunting dogs showed elevated antibody titers to Francisella tularensis , but the ... ...

    Abstract In November 2018, an outbreak of tularemia occurred among hare hunters in Bavaria, Germany. At least one infected hare was confirmed as the source of infection. A number of hunting dogs showed elevated antibody titers to Francisella tularensis , but the absence of titer increases in subsequent samples did not point to acute infections in dogs. Altogether, 12 persons associated with this hare hunt could be diagnosed with acute tularemia by detection of specific antibodies. In nine patients, the antibody and cytokine responses could be monitored over time. Eight out of these nine patients had developed detectable antibodies three weeks after exposure; in one individual the antibody response was delayed. All patients showed an increase in various cytokines and chemokines with a peak for most mediators in the first week after exposure. Cytokine levels showed individual variations, with high and low responders. The kinetics of seroconversion has implications on serological diagnoses of tularemia.
    Keywords bacterial infections ; tularemia ; hares ; Francisella ; Francisella tularensis ; outbreak ; Biology (General) ; QH301-705.5
    Language English
    Publishing date 2020-10-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  7. Article: Comparative analyses of Legionella species identifies genetic features of strains causing Legionnaires’ disease

    Gomez-Valero, Laura / Carmen Buchrieser / Christophe Rusniok / Claudine Médigue / Delphine Dervins-Ravault / Elizabeth L Hartland / Gernot Glöckner / Honglei Chen / Jasmin Demirtas / Jerome Etienne / Klaus Heuner / Mario Neou / Michael Steinert / Monica Rolando / Nicola K Petty / Robert J Moore / Simonetta Gribaldo / Sophie Jarraud / Zoe Rouy

    Genome biology. 2014 Nov., v. 15, no. 11

    2014  

    Abstract: BACKGROUND: The genus Legionella comprises over 60 species. However, L. pneumophila and L. longbeachae alone cause over 95% of Legionnaires’ disease. To identify the genetic bases underlying the different capacities to cause disease we sequenced and ... ...

    Abstract BACKGROUND: The genus Legionella comprises over 60 species. However, L. pneumophila and L. longbeachae alone cause over 95% of Legionnaires’ disease. To identify the genetic bases underlying the different capacities to cause disease we sequenced and compared the genomes of L. micdadei, L. hackeliae and L. fallonii (LLAP10), which are all rarely isolated from humans. RESULTS: We show that these Legionella species possess different virulence capacities in amoeba and macrophages, correlating with their occurrence in humans. Our comparative analysis of 11 Legionella genomes belonging to five species reveals highly heterogeneous genome content with over 60% representing species-specific genes; these comprise a complete prophage in L. micdadei, the first ever identified in a Legionella genome. Mobile elements are abundant in Legionella genomes; many encode type IV secretion systems for conjugative transfer, pointing to their importance for adaptation of the genus. The Dot/Icm secretion system is conserved, although the core set of substrates is small, as only 24 out of over 300 described Dot/Icm effector genes are present in all Legionella species. We also identified new eukaryotic motifs including thaumatin, synaptobrevin or clathrin/coatomer adaptine like domains. CONCLUSIONS: Legionella genomes are highly dynamic due to a large mobilome mainly comprising type IV secretion systems, while a minority of core substrates is shared among the diverse species. Eukaryotic like proteins and motifs remain a hallmark of the genus Legionella. Key factors such as proteins involved in oxygen binding, iron storage, host membrane transport and certain Dot/Icm substrates are specific features of disease-related strains.
    Keywords clathrin ; genes ; humans ; Legionella ; macrophages ; oxygen ; physiological transport ; secretion ; type IV secretion system ; virulence
    Language English
    Dates of publication 2014-11
    Size p. 505.
    Publishing place Springer-Verlag
    Document type Article
    ZDB-ID 2040529-7
    ISSN 1474-760X ; 1465-6914 ; 1465-6906
    ISSN (online) 1474-760X ; 1465-6914
    ISSN 1465-6906
    DOI 10.1186/s13059-014-0505-0
    Database NAL-Catalogue (AGRICOLA)

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