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21 pages, 2506 KiB

Open AccessArticle

Reorganized Genomic Taxonomy of Francisellaceae Enables Design of Robust Environmental PCR Assays for Detection of Francisella tularensis

by Caroline Öhrman, Jason W. Sahl, Andreas Sjödin, Ingrid Uneklint, Rebecca Ballard, Linda Karlsson, Ryelan F. McDonough, David Sundell, Kathleen Soria, Stina Bäckman, Kitty Chase, Björn Brindefalk, Shanmuga Sozhamannan, Adriana Vallesi, Emil Hägglund, Jose Gustavo Ramirez-Paredes, Johanna Thelaus, Duncan Colquhoun, Kerstin Myrtennäs, Dawn Birdsell, Anders Johansson, David M. Wagner and Mats Forsman Show full author list Hide full author list

Microorganisms 2021, 9(1), 146; https://doi.org/10.3390/microorganisms9010146 - 11 Jan 2021

Cited by 19 | Viewed by 5066

Abstract

In recent years, an increasing diversity of species has been recognized within the family Francisellaceae. Unfortunately, novel isolates are sometimes misnamed in initial publications or multiple sources propose different nomenclature for genetically highly similar isolates. Thus, unstructured and occasionally incorrect information can [...] Read more.

In recent years, an increasing diversity of species has been recognized within the family Francisellaceae. Unfortunately, novel isolates are sometimes misnamed in initial publications or multiple sources propose different nomenclature for genetically highly similar isolates. Thus, unstructured and occasionally incorrect information can lead to confusion in the scientific community. Historically, detection of Francisella tularensis in environmental samples has been challenging due to the considerable and unknown genetic diversity within the family, which can result in false positive results. We have assembled a comprehensive collection of genome sequences representing most known Francisellaceae species/strains and restructured them according to a taxonomy that is based on phylogenetic structure. From this structured dataset, we identified a small number of genomic regions unique to F. tularensis that are putatively suitable for specific detection of this pathogen in environmental samples. We designed and validated specific PCR assays based on these genetic regions that can be used for the detection of F. tularensis in environmental samples, such as water and air filters. Full article

(This article belongs to the Special Issue Epidemiology of Tularemia and Francisella tularensis)

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12 pages, 792 KiB

Open AccessArticle

Genotyping of Francisella tularensis subsp. holarctica from Hares in Germany

by Jörg Linde, Timo Homeier-Bachmann, Alexandra Dangel, Julia M. Riehm, David Sundell, Caroline Öhrman, Mats Forsman and Herbert Tomaso

Microorganisms 2020, 8(12), 1932; https://doi.org/10.3390/microorganisms8121932 - 5 Dec 2020

Cited by 8 | Viewed by 2476

Abstract

Francisella tularensis is the causative agent of the zoonotic disease tularemia. In Germany, most human infections are caused by contact with infected hares. The aim of this study was to characterize Francisella tularensis subsp. holarctica strains isolated from hares in Germany and to [...] Read more.

Francisella tularensis is the causative agent of the zoonotic disease tularemia. In Germany, most human infections are caused by contact with infected hares. The aim of this study was to characterize Francisella tularensis subsp. holarctica strains isolated from hares in Germany and to develop bioinformatics tools to analyze their genetic relatedness. In total, 257 German isolates—obtained mainly from hares (n = 233), other vertebrate animals, and ticks, but also from humans (n = 3)—were analyzed within this study. Publically available sequence data from 49 isolates were used to put our isolates into an epidemiological context and to compare isolates from natural foci and humans. Whole-genome sequences were analyzed using core-genome Multi-Locus-Sequence-Typing, canonical Single Nucleotide Polymorphism (SNP) typing and whole-genome SNP typing. An overall conformity of genotype clustering between the typing methods was found, albeit with a lower resolution for canonical single SNP typing. The subclade distribution, both on local and national levels, among strains from humans and hares was similar, suggesting circulation of the same genotypes both in animals and humans. Whilst close to identical isolates of the same subclade were found distributed over large areas, small geographical foci often harbored members of different subclades. In conclusion, although genomic high-resolution typing was shown to be robust, reproducible and allowed the identification of highly closely related strains, genetic profiling alone is not always conclusive for epidemiological linkage of F. tularensis strains. Full article

(This article belongs to the Special Issue Epidemiology of Tularemia and Francisella tularensis)

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14 pages, 1617 KiB

Open AccessArticle

Genetic Traces of the Francisella tularensis Colonization of Spain, 1998–2020

by Kerstin Myrtennäs, Raquel Escudero, Ángel Zaballos, Rosa González-Martín-Niño, Miklós Gyuranecz and Anders Johansson

Microorganisms 2020, 8(11), 1784; https://doi.org/10.3390/microorganisms8111784 - 14 Nov 2020

Cited by 2 | Viewed by 2130

Abstract

More than 1000 humans have acquired the febrile disease tularemia in Spain since the first notification of human cases in 1997. We here aimed to study the recent molecular evolution of the causative bacterium Francisella tularensis during disease establishment in Spain. Single-nucleotide polymorphisms [...] Read more.

More than 1000 humans have acquired the febrile disease tularemia in Spain since the first notification of human cases in 1997. We here aimed to study the recent molecular evolution of the causative bacterium Francisella tularensis during disease establishment in Spain. Single-nucleotide polymorphisms (SNPs) and variable-number tandem repeats (VNTRs) were analyzed in whole-genome sequences (WGS) of F. tularensis. Short-read WGS data for 20 F. tularensis strains from humans infected in the periods 2014–2015 and 2018–2020 in Spain were generated. These data were combined with WGS data of 25 Spanish strains from 1998 to 2008 and two reference strains. Capillary electrophoresis data of VNTR genetic regions were generated and compared with the WGS data for the 11 strains from 2014 to 2015. Evolutionary relationships among strains were analyzed by phylogenetic methods. We identified 117 informative SNPs in a 1,577,289-nucleotide WGS alignment of 47 F. tularensis genomes. Forty-five strains from Spain formed a star-like SNP phylogeny with six branches emerging from a basal common node. The most recently evolved genomes formed four additional star-like structures that were derived from four branches of the basal common node. VNTR copy number variation was detected in two out of 10 VNTR regions examined. Genetic clustering of strains by VNTRs agreed with the clustering by SNPs. The SNP data provided higher resolution among strains than the VNTRs data in all but one cases. There was an excellent correlation between VNTR marker sizing by capillary electrophoresis and prediction from WGS data. The genetic data strongly support that tularemia, indeed, emerged recently in Spain. Distinct genetic patterns of local F. tularensis population expansions imply that the pathogen has colonized a previously disease-free geographical area. We also found that genome-wide SNPs provide higher genetic resolution among F. tularensis genomes than the use of VNTRs, and that VNTR copy numbers can be accurately predicted using short-read WGS data. Full article

(This article belongs to the Special Issue Epidemiology of Tularemia and Francisella tularensis)

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10 pages, 1383 KiB

Open AccessCommunication

Differences in Blood-Derived Francisella tularensis Type B Strains from Clinical Cases of Tularemia

by Marilynn A. Larson, Baha Abdalhamid, Bhanwar Lal Puniya, Tomáš Helikar, David W. Kelley and Peter C. Iwen

Microorganisms 2020, 8(10), 1515; https://doi.org/10.3390/microorganisms8101515 - 1 Oct 2020

Cited by 3 | Viewed by 2137

Abstract

Francisella tularensis can cause the zoonotic disease tularemia and is partitioned into subspecies due to differences in chromosomal organization and virulence. The subspecies holarctica (type B) is generally considered more clonal than the other subpopulations with moderate virulence compared to the hypervirulent A.I [...] Read more.

Francisella tularensis can cause the zoonotic disease tularemia and is partitioned into subspecies due to differences in chromosomal organization and virulence. The subspecies holarctica (type B) is generally considered more clonal than the other subpopulations with moderate virulence compared to the hypervirulent A.I clade. We performed whole genome sequencing (WGS) on six type B strains isolated from the blood of patients with tularemia within a one-year period from the same United States region, to better understand the associated pathogenicity. The WGS data were compared to the prototype strain for this subspecies, specifically FSC200, which was isolated from a patient with tularemia in Europe. These findings revealed 520–528 single nucleotide polymorphisms (SNPs) between the six United States type B strains compared to FSC200, with slightly higher A+T content in the latter strain. In contrast, comparisons between the six type B isolates showed that five of the six type B isolates had only 4–22 SNPs, while one of the strains had 47–53 SNPs. Analysis of SNPs in the core genome for the six United States type B isolates and the FSC200 strain gave similar results, suggesting that some of these mutations may have been nonsynonymous, resulting in altered protein function and pathogenicity. Full article

(This article belongs to the Special Issue Epidemiology of Tularemia and Francisella tularensis)

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15 pages, 2112 KiB

Open AccessArticle

The Comparative Virulence of Francisella tularensis Subsp. mediasiatica for Vaccinated Laboratory Animals

by Vitalii Timofeev, Galina Titareva, Irina Bahtejeva, Tatiana Kombarova, Tatiana Kravchenko, Alexander Mokrievich and Ivan Dyatlov

Microorganisms 2020, 8(9), 1403; https://doi.org/10.3390/microorganisms8091403 - 12 Sep 2020

Cited by 8 | Viewed by 1965

Abstract

Tularemia is a severe infectious disease caused by the Gram-negative bacteria Fracisella tularensis. There are four subspecies of F.tularensis: holarctica, tularensis, mediasiatica, and novicida, which differ in their virulence and geographic distribution. One of them, subsp. mediasiatica [...] Read more.

Tularemia is a severe infectious disease caused by the Gram-negative bacteria Fracisella tularensis. There are four subspecies of F.tularensis: holarctica, tularensis, mediasiatica, and novicida, which differ in their virulence and geographic distribution. One of them, subsp. mediasiatica remains extremely poorly studied, primarily due to the fact that it is found only in the sparsely populated regions of Central Asia and Russia. In particular there is little information in the literature on the virulence and pathogenicity of subsp. mediasiatica. In the present article, we evaluated the comparative virulence of subsp. mediasiatica in vaccinated laboratory animals which we infected with virulent strains: subsp. mediasiatica 678, subsp. holarctica 503, and subsp. tularensis SCHU within 60 to 180 days after vaccination. We found that subsp. mediasiatica is comparable in pathogenicity in mice with subsp. tularensis and in guinea pigs with subsp. holarctica. We also found that the live vaccine does not fully protect mice from subsp. mediasiatica but completely protects guinea pigs for at least six months. In general, our data suggest that subsp. mediasiatica occupies an intermediate position in virulence between spp. tularensis and holarctica. Full article

(This article belongs to the Special Issue Epidemiology of Tularemia and Francisella tularensis)

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Review

Jump to: Research

18 pages, 690 KiB

Open AccessReview

Mechanisms Affecting the Acquisition, Persistence and Transmission of Francisella tularensis in Ticks

by Brenden G. Tully and Jason F. Huntley

Microorganisms 2020, 8(11), 1639; https://doi.org/10.3390/microorganisms8111639 - 23 Oct 2020

Cited by 11 | Viewed by 6094

Abstract

Over 600,000 vector-borne disease cases were reported in the United States (U.S.) in the past 13 years, of which more than three-quarters were tick-borne diseases. Although Lyme disease accounts for the majority of tick-borne disease cases in the U.S., tularemia cases have been [...] Read more.

Over 600,000 vector-borne disease cases were reported in the United States (U.S.) in the past 13 years, of which more than three-quarters were tick-borne diseases. Although Lyme disease accounts for the majority of tick-borne disease cases in the U.S., tularemia cases have been increasing over the past decade, with >220 cases reported yearly. However, when comparing Borrelia burgdorferi (causative agent of Lyme disease) and Francisella tularensis (causative agent of tularemia), the low infectious dose (<10 bacteria), high morbidity and mortality rates, and potential transmission of tularemia by multiple tick vectors have raised national concerns about future tularemia outbreaks. Despite these concerns, little is known about how F. tularensis is acquired by, persists in, or is transmitted by ticks. Moreover, the role of one or more tick vectors in transmitting F. tularensis to humans remains a major question. Finally, virtually no studies have examined how F. tularensis adapts to life in the tick (vs. the mammalian host), how tick endosymbionts affect F. tularensis infections, or whether other factors (e.g., tick immunity) impact the ability of F. tularensis to infect ticks. This review will assess our current understanding of each of these issues and will offer a framework for future studies, which could help us better understand tularemia and other tick-borne diseases. Full article

(This article belongs to the Special Issue Epidemiology of Tularemia and Francisella tularensis)

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13 pages, 604 KiB

Open AccessReview

Tularemia Goes West: Epidemiology of an Emerging Infection in Austria

by Stefanie Seiwald, Anja Simeon, Erwin Hofer, Günter Weiss and Rosa Bellmann-Weiler

Microorganisms 2020, 8(10), 1597; https://doi.org/10.3390/microorganisms8101597 - 16 Oct 2020

Cited by 22 | Viewed by 3184

Abstract

The zoonotic disease tularemia is caused by the Gram-negative bacterium Francisella tularensis, with the two major subspecies tularensis and holarctica being responsible for infections in humans and animals. The F. tularensis subspecies holarctica is less virulent and prevalent in Europe and Asia. [...] Read more.

The zoonotic disease tularemia is caused by the Gram-negative bacterium Francisella tularensis, with the two major subspecies tularensis and holarctica being responsible for infections in humans and animals. The F. tularensis subspecies holarctica is less virulent and prevalent in Europe and Asia. Over the last few centuries, few epidemic outbreaks and low numbers of infections have been registered in the eastern part of Austria, specifically in the provinces of Lower Austria, Burgenland, and Styria. The reported infections were mostly associated with hunting hares and the skinning of carcasses. Within the last decade, ticks have been identified as important vectors in Tyrol and served as first evidence for the spread of F. tularensis to Western Austria. In 2018, the pathogen was detected in hares in the provinces of Tyrol, Vorarlberg, and Salzburg. We presume that F. tularensis is now established in most regions of Austria, and that the investigation of potential host and vector animals should be spotlighted by public institutions. Tularemia in humans presents with various clinical manifestations. As glandular, ulceroglandular, and typhoidal forms occur in Austria, this infectious disease should be considered as a differential diagnosis of unknown fever. Full article

(This article belongs to the Special Issue Epidemiology of Tularemia and Francisella tularensis)

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14 pages, 2747 KiB

Open AccessReview

Francisella tularensis Subspecies holarctica and Tularemia in Germany

by Sandra Appelt, Mirko Faber, Kristin Köppen, Daniela Jacob, Roland Grunow and Klaus Heuner

Microorganisms 2020, 8(9), 1448; https://doi.org/10.3390/microorganisms8091448 - 22 Sep 2020

Cited by 21 | Viewed by 3967

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 [...] Read more.

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. Full article

(This article belongs to the Special Issue Epidemiology of Tularemia and Francisella tularensis)

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13 pages, 5086 KiB

Open AccessReview

Epidemiologic and Epizootic Data of Tularemia in the Past and in the Recent History in Croatia

by Mirna Mihelčić, Valentina Marečić, Mateja Ožanič, Ina Kelava, Maša Knežević and Marina Šantić

Microorganisms 2020, 8(5), 721; https://doi.org/10.3390/microorganisms8050721 - 12 May 2020

Cited by 2 | Viewed by 2624

Abstract

Tularemia is a zoonotic disease caused by Francisella tularensis. A large number of recent studies have provided an update on the disease characteristics and the distribution across Europe. In Croatia, most of the clinical cases, as well as the reports of the [...] Read more.

Tularemia is a zoonotic disease caused by Francisella tularensis. A large number of recent studies have provided an update on the disease characteristics and the distribution across Europe. In Croatia, most of the clinical cases, as well as the reports of the disease in animals, date from the 20th century. In that period, epidemic and epizootic research had given detailed information about endemic regions and their characteristics, including suspected animal hosts and vectors. The region along the middle course of the Sava River, called Middle Posavina, is described as an endemic region, i.e., a “natural focus” of tularemia, in Croatia. In the 21st century, cases of human tularemia are being reported sporadically, with ulceloglandular, oropharyngeal and typhoid forms of disease. A majority of the described cases are linked with the consumption of contaminated food or water. The disease outbreaks still occur in areas along the course of the river Sava and in northwest Croatia. In this review article, we have summarized epidemiologic and epizootic data of tularemia in the past and in recent Croatian history. Full article

(This article belongs to the Special Issue Epidemiology of Tularemia and Francisella tularensis)

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