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The C. Comparison of these two chromosomes with three available reference C. Some of these divergent genomic regions in both chromosomes are phage- and plasmid-related segments.

Five of these shared regions formed a mosaic of plasmid-integrated segments, suggesting that these elements were acquired early in a clonal lineage of netF-positive C.

A comparative analysis of tcp -conjugative plasmids has shown that these plasmids share a highly conserved 35 kb core region and a diverse variable region.

A general feature of C. The present study describes the complete genome sequence of two netF -positive C.

The particular emphasis of the current study is on the plasmids shared by these two netF -positive strains. Two netF -positive type A C.

These isolates were selected on the basis of their clonal relationship identified in a previous study [ 6 ]. The quality of the genomic DNA was evaluated by standard agarose gel electrophoresis and the identity as the correct bacterium confirmed by PCR amplification of cpa , cpe , netE , netF , and netG.

This strategy provided an opportunity to successfully close the genome sequences of these two samples and the necessary accuracy of base calls for the sequences.

Assembly errors and poor quality data were manually trimmed where possible. The contigs were oriented and ordered according to the closed C.

The chromosomal unique regions were identified by comparison to three complete C. The Pan-genome Analyses of PanSeq software was used to determine the core sequence of conjugative plasmids of this study by comparison to the same six complete C.

The PacBio-SMRT sequencing technology generated 28, and 41, reads, with a raw median read length of 4, and 5, bp, totalling ,, fold coverage and ,, fold coverage nucleotides for JFP55 and JFP, respectively.

For JFP55, this assembly produced 64 contigs minimum contig length: and maximum contig length: 2,, whereas for JFP 46 contigs minimum contig length: and maximum contig length: 3,, were generated.

Furthermore, the assembled chromosomes were compared to the closed chromosome of ATCC using progressiveMauve software to assess the validity of the assemblies.

A summary of the genome assembly results is presented in Table 1. Plasmid annotation revealed that both netF -positive C. The common backbone region contains 22 genes, which encode constituents of the tcp locus tcpACDEFGHIJ , a plasmid replication gene rep , a DNA-binding transcriptional repressor regD , a growth inhibitor PemK protein, a sortase, a DNA adenine-specific methyltransferase dam , a tyrosine site-specific recombinase, and seven hypothetical proteins with unknown functions.

The arrows with no name are hypothetical proteins. The image was generated using Easyfig [ 28 ]. This locus encoded two putative beta-channel pore-forming toxins, NetF and NetE, and other predicted proteins, which possibly contribute to the pathogenesis of netF -positive C.

These proteins contained a Cna-like B-region collagen-binding protein domain, and a gene encoding a sortase enzyme was located immediately downstream.

Analysis of the genomic inversion indicated long bp and nearly perfect inverted repeat sequences near its termini —, — In addition, the inversion was confirmed by PCR and by sequencing of each amplicon data not shown.

Plasmid pJFP55G is an incomplete sequence. The image was generated using Easyfig. The annotation revealed the presence of 17 CDSs in each plasmid.

Three hundred and forty-four Sequence analysis with BLASTN indicated that this plasmid is a unique mega-plasmid with virtually no homology to other clostridia sequences in GenBank.

No genes with apparent relationship to direct virulence functions were identified. Thirty-nine of these This locus encodes NetG, a putative beta-sheet pore-forming toxin, as well as 33 additional predicted proteins, 19 with unknown functions Table 4 and Fig 1.

Visual comparative analysis of the two netF -positive C. In terms of chromosome size, JFP55 and JFP are slightly larger than the three other closed chromosomes and carry a number of unique regions Fig 3.

Starting from the outermost ring the feature rings depict: 1. Forward strand coding sequence of JFP; 2.

Reverse strand coding sequence of JFP; 3. JFP55; 4. ATCC; 5. Strain 13; 6. The last two rings display the GC content and GC skew.

The features of those unique to the two netF -positive strains in comparison to the three reference genomes but shared between JFP55 and JFP are presented in S9 Table.

No shared virulence genes unique to the two netF -positive chromosomes were identified. Interestingly, five of these shared regions were part of a larger region, split into these five small sub-regions.

These include, for example, CPE enterotoxin-producing strains associated with food poisoning in humans [ 33 ] or with antibiotic-associated diarrhea in humans [ 34 ].

Most recently, the pathotype diversity has been highlighted by the discovery of the large conjugative plasmid-encoded toxin NetF, its association with canine hemorrhagic gastroenteritis and foal necrotizing enteritis, and the common clonal lineage of these isolates [ 6 ].

It is clear that the traditional toxinotyping scheme requires modifications to include these new findings, and to adapt to the diversity of distinct enteric disease caused by this bacterium.

The current study provides the first complete genome sequences of two netF -positive C. Considerable work remains to be done to understand the contributions of the novel genes and loci identified in this study.

In relation to pathogenic C. A previous paper [ 11 ] described regions unique to the netB -pathotype as pathogenicity loci PaLoc.

We found that these NetF-producing C. This finding suggests that the key event in the evolution of netF -positive C. The hypothesis of a key evolutionary event is further supported by the previous finding that the presence of NetF is crucial for producing cytotoxicity in vitro [ 6 ].

The 20 kb inversion of the region of the NetF pathogenicity locus containing the netE and netF genes suggests the mechanism by which this pathogenicity locus added the critically important netF toxin gene [ 6 ], since this region is flanked by a large inverted repeat.

Acquisition of this region was likely an important event in the evolution of this virulence plasmid and of this C.

An interesting trait of the netF -positive strains is that they always contain a CPE-bearing plasmid [ 6 ]. The consistent presence of cpe plasmid in these strains suggests that the ancestral strain also possessed this plasmid, or acquired it early in stages of expansion of this lineage, and also that CPE production may be important in the pathogenesis of disease caused by netF -positive strains [ 6 ].

Recently, Uzal and others [ 37 ] have demonstrated a synergistic effect of CPB and CPE of a type C human enteritis necroticans strain in producing histological damage and fluid accumulation in rabbit intestinal loops.

It has been shown that the production of bacteriocins is a common feature of C. The presence of the closely related bacteriocin plasmid in both NetF-producing strains suggests its importance in this lineage.

Apart from the common plasmids in both netF -positive strains, the genome of each strain contains two unique plasmids. One unique plasmid of interest is the mega-plasmid, pJFPA.

Although mega-plasmids are a common feature in some clostridal species, such as neurotoxigenic C.

The variable presence of these unique plasmids in NetF-producing strains suggests that these have been acquired during evolution from the ancestral strains and may not be important in virulence.

The inconsistent presence of this putative toxin gene in netF -positive C. The toxin-encoding plasmids described in the current study are members of tcp -conjugative family plasmids.

These plasmids encode the tcp locus, which shares minor sequence homology with Tn conjugative transposon family [ 19 ].

It is therefore likely that these are conjugative plasmids but we did not explore this and this still needs to be tested in conjugation experiments.

For instance, the gene encoding NetB is localized downstream of the conserved dcm region on conjugative variably-sized plasmids 80—90 kb [ 9 , 16 , 35 ].

Interesting features of this include an internalin A-like protein, as well as, two putative cell surface adhesion proteins.

The internalin family was originally identified in Listeria monocytogenes as cell surface proteins which mediate the bacterial adhesion and invasion [ 42 ].

In some Clostridium species such as C. In the NetB pathogenicity locus, a putative internalin-like protein was also found immediately upstream from netB gene [ 11 ].

While the role of these internalin-like proteins has not yet been fully defined, the presence of leucine-rich repeats domains suggests that they are likely involved in protein-protein interaction [ 45 ].

These surface proteins contained a Cna-like B-region domain, which is originally found in the Staphylococcus aureus collagen-binding protein where it acts as a stalk to present the ligand-binding domain of adhesion away from the bacterial cell surface [ 46 ].

This domain was found in the Streptococcus pneumoniae pilus protein, RrgB, and acts in many Gram-positive surface proteins either as pilin subunit cross-linking or cell wall attachment [ 47 ].

Further functional studies are required to elucidate the possible contributions of these proteins in bacterial attachment to the host cell surface.

The cpe plasmids of type A C. The cpe gene on both types of plasmids is flanked by an upstream IS sequence.

A previous study in Clostridium difficile has shown that the holin-like protein, TcdE, is required for export of the enterotoxins TcdA and TcdB [ 51 ].

Whether the holin-like protein found in the enterotoxin locus of both pJFP55G and pJFPD plays a role in exporting of enterotoxin remains to be investigated.

As noted, the variable presence of netG is a feature of NetF-producing strains. This protein contains two domains, the peptidase Mlike superfamily E value: 4E and discoidin family domain E value: 2E The Mlike superfamily contains a zinc metallopeptidase shown to be involved in mucinase activity [ 52 ].

In addition, proteins containing discoidin domains are predicted to bind carbohydrates such as galactose [ 53 ].

An intriguing hypothesis is that this protein may be involved in mucin colonization of C. This finding suggests that these NetF-producing strains harbor chromosomal unique regions missing in the three reference strains.

The novel region finder of PanSeq tool identified regions unique to each of the chromosome of JFP55 and JFP, respectively and absent from the chromosome of three references strains.

Large unique regions included complete and partial phage sequences, as well as regions likely associated with capsule formation. These regions have some classic hallmarks of plasmid genes, such as the collagen-binding protein first identified in pCP13 by Shimizu and others [ 7 ].

Although it is well known that the enterotoxin gene cpe can move between plasmid and chromosome of C. We found that only 16 unique regions were shared by two netF -positive C.

Five of these common regions formed a mosaic of plasmid-integrated segments. These five regions are adjacent and likely originate from a single integration event followed by recombination.

This finding suggests that these elements were acquired early in a clonal lineage of netF -positive C.

In addition, the presence of multiple chromosomal unique regions, which are not shared by the two netF -positive strains suggests these strains subsequently diverged for an extended time.

Further work is required to assess the significance of chromosomal regions unique to NetF-producing C.

In addition to this chromosomally encoded adherence factor, we found three and two other collagen binding proteins on the tcp -conjugative plasmids of JFP and JFP55, respectively.

The presence of this number of adhesin genes is intriguing and suggests a possible role in the intestinal colonization of netF -positive strains.

In summary, we found that the JFP55 and JFP strains, which originated from foal necrotizing enteritis and canine hemorrhagic gastroenteritis cases, share unique virulence genes on conserved pathogenicity loci found on large tcp -conjugative plasmids.

The identification of common features for these two strains provides supportive evidence that these two netF -positive strains are a part of a common clonal lineage [ 6 ].

Moreover, these results provide significant insight into the potential pathogenesis basis of canine and foal necrotizing enteritis and into the evolution of virulence of C.

Read article at publisher's site DOI : Pathogens , 8 1 , 29 Jan J Vet Intern Med , 33 1 , 30 Nov Front Microbiol , , 14 Aug BMC Genomics , 19 1 , 22 May Front Microbiol , , 12 Dec The current study presents the complete genome sequence of two netF-positive strains, JFP55 and JFP, which were recovered from cases of foal necrotizing enteritis and canine hemorrhagic gastroenteritis, respectively.

The putative beta-pore-forming toxin genes, netF, netE and netG, were located in unique pathogenicity loci on tcp-conjugative plasmids.

The C. Comparison of these two chromosomes with three available reference C.

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CCT map comparing the chromosomes of two netF -positive C. Table 7 General features of unique regions identified by PanSeq. Discussion The current typing system for C.

XLSX Click here for additional data file. S9 Table Shared unique chromosomal nucleotide sequences by two netF -positive C.

Data Availability All relevant data are within the paper and its Supporting Information files. References 1. Songer JG. Clostridial enteric diseases of domestic animals.

Clin Microbiol Rev. The prokaryotes: a handbook on the biology of bacteria. Springer; Hatheway C. Toxigenic clostridia.

Clostridium perfringens : toxinotype and genotype. Trends Microbiol. Clostridium Perfringens toxins involved in mammalian veterinary diseases.

Open Toxinology J. A novel pore-forming toxin in type A Clostridium perfringens is associated with both fatal canine hemorrhagic gastroenteritis and fatal foal necrotizing enterocolitis.

April 8; 10 4 :e Complete genome sequence of Clostridium perfringens , an anaerobic flesh-eater.

Proc Natl Acad Sci. Skewed genomic variability in strains of the toxigenic bacterial pathogen, Clostridium perfringens. Genome Res.

Sequence of two plasmids from Clostridium perfringens chicken necrotic enteritis isolates and comparison with C.

November 26; 7 11 : e Genome analyses of Clostridium perfringens isolates from five toxinotypes. Res Microbiol. Identification of novel pathogenicity loci in Clostridium perfringens strains that cause avian necrotic enteritis.

May 24; 5 5 : e Toxin plasmids of Clostridium perfringens. Microbiol Mol Biol Rev. BEC, a novel enterotoxin of Clostridium perfringens found in human clinical isolates from acute gastroenteritis outbreaks.

Infect Immun. Enterotoxin plasmid from Clostridium perfringens is conjugative. Epsilon-toxin plasmids of Clostridium perfringens type D are conjugative.

J Bacteriol. Necrotic enteritis-derived Clostridium perfringens strain with three closely related independently conjugative toxin and antibiotic resistance plasmids.

September 27; 2 5 : e— Identification of a transferable tetracycline resistance plasmid pCW3 from Clostridium perfringens.

Worldwide distribution of the conjugative Clostridium perfringens tetracycline resistance plasmid, pCW3.

Functional identification of conjugation and replication regions of the tetracycline resistance plasmid pCW3 from Clostridium perfringens.

The peptidoglycan hydrolase TcpG is required for efficient conjugative transfer of pCW3 in Clostridium perfringens. The conjugation protein TcpC from Clostridium perfringens is structurally related to the type IV secretion system protein VirB8 from Gram-negative bacteria.

Mol Microbiol. Sequencing and diversity analyses reveal extensive similarities between some epsilon-toxin-encoding plasmids and the pCPF Clostridium perfringens enterotoxin plasmid.

Pospiech A, Neumann B. A versatile quick-prep of genomic DNA from Gram-positive bacteria. Trends Genet. June 25; 5 6 : e Nucleic Acids Res.

January 1; 42 : D—D Basic local alignment search tool. J Mol Biol. PHAST: a fast phage search tool. July 1; 7 39 :W— Easyfig: a genome comparison visualizer.

April 1; 27 7 : — The spatial organization of the VirR boxes is critical for VirR-mediated expression of the perfringolysin O gene, pfoA , from Clostridium perfringens.

Stothard P, Wishart DS. Circular genome visualization and exploration using CGView. McClane BA. An overview of Clostridium perfringens enterotoxin.

Genotyping of enterotoxigenic Clostridium perfringens fecal isolates associated with antibiotic-associated diarrhea and food poisoning in North America.

J Clin Microbiol. NetB, a new toxin that is associated with avian necrotic enteritis caused by Clostridium perfringens.

PLoS Pathog. February 8; 4 2 : e26 Multilocus sequence typing analysis of Clostridium perfringens isolates from necrotic enteritis outbreaks in broiler chicken populations.

Towards an understanding of the role of Clostridium perfringens toxins in human and animal disease. Future Microbiol. Bacteriocins of Gram-positive bacteria.

Necrotic enteritis-producing strains of Clostridium perfringens displace non-necrotic enteritis strains from the gut of chicks.

Vet Microbiol. Structure and genetic content of the megaplasmids of neurotoxigenic Clostridium butyricum type E strains from Italy.

August 15; 8 8 :e Complete nucleotide sequence of a plasmid containing the botulinum neurotoxin gene in Clostridium botulinum type B strain isolated from an infant patient in Japan.

Mol Genet Genomics. Listeria monocytogenes : a multifaceted model. Nat Rev Micro. The genome sequence of Clostridium tetani , the causative agent of tetanus disease.

Genome sequence of a proteolytic Group I Clostridium botulinum strain Hall A and comparative analysis of the clostridial genomes.

The leucine-rich repeat structure. Cell Mol Life Sci. Novel fold and assembly of the repetitive B region of the Staphylococcus aureus collagen-binding surface protein.

Supramolecular organization of the repetitive backbone unit of the Streptococcus pneumonia pilus. June 15; 5 6 :e Organization of the plasmid cpe locus of Clostridium perfringens type A isolates.

Complete sequencing and diversity analysis of the enterotoxin-encoding plasmids in Clostridium perfringens type A non-food-borne human gastrointestinal disease isolates.

June 3; 5 6 : e Govind R, Dupuy B. June 7; 8 6 :e A novel extracellular metallopeptidase domain shared by animal host-associated mutualistic and pathogenic microbes.

January 27; 7 1 :e The three domains of a bacterial sialidase: a beta-propeller, an immunoglobulin module and a galactose-binding jelly-roll.

The enterotoxin gene cpe of Clostridium perfringens can be chromosomal or plasmid-borne. Novel insights into the epidemiology of Clostridium perfringens type A food poisoning.

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March 8; 7 3 :e Smart citations by scite. The number of the statements may be higher than the number of citations provided by EuropePMC if one paper cites another multiple times or lower if scite has not yet processed some of the citing articles.

Explore citation contexts and check if this article has been supported or disputed. Prevalence of Clostridium perfringens netE and netF toxin genes in the feces of dogs with acute hemorrhagic diarrhea syndrome.

Whole genome analysis reveals the diversity and evolutionary relationships between necrotic enteritis-causing strains of Clostridium perfringens.

BioStudies: supplemental material and supporting data. Nucleotide Sequences Showing 20 of Show less.

Data that cites the article This data has been provided by curated databases and other sources that have cited the article.

The putative beta-pore-forming toxin genes, netF, netE and netG, were located in unique pathogenicity loci on tcp-conjugative plasmids.

The C. Comparison of these two chromosomes with three available reference C. Some of these divergent genomic regions in both chromosomes are phage- and plasmid-related segments.

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See https://duffyboats.se/filme-schauen-stream/butter-bei-die-fische-film.php. As noted, the variable presence of netG is a feature of NetF-producing strains. September 27; 2 5 : e— Weese1 Valeria R. Explore citation contexts and check if this article has been supported or disputed. By continuing to browse this bad moms, you agree to allow omicX and its partners to use cookies to click here the site's operation and effectiveness, to display ads tailored to your interests and to provide you with relevant promotional messages and other information about products, events and services of ours or our sponsors and partner companies. Smart citations by scite. Free ritter jedi. Our Privacy Continue reading outlines how sprung leben use cookies and sam alte folgen technologies.

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Hypothetical protein A. Scott J. Abstract The recent discovery of a novel beta-pore-forming toxin, NetF, which is strongly associated with canine and foal continue reading enteritis should improve our understanding here the role of type A Clostridium perfringens associated disease in these animals. March 8; 7 3 :e Seller does not offer returns. Clostridium perfringens : toxinotype and genotype. Nach der 2. Zurück zur Startseite Zurück zum Seitenanfang. Nehmen Sie Netflix einfach überall hin mit. Den ganzen Artikel lesen: "Snowpiercer": Wann erscheinen neue Fuck the Staffel der Comedy-Serie? EUR 21, Staffel auf Netflix? Mehr erfahren: "The Sinner" auf Netflix: Wird es eine 4. Es gibt immer etwas Neues zu entdecken und jede Woche kommen weitere Serien und Filme dazu. Bitte geben Sie für spielfilme deutsch voller lпїЅnge Postleitzahl fünf oder neun Ziffern ein. Ihre Beobachtungsliste ist more info. Netflix enthüllt, wann es weitergeht. Wir verwenden Cookies warum? La Casa Di Carta. Staffel auf Netflix erscheinen? Verkäufer kontaktieren.

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