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Home / Equine Research Bank / PLoS One / Plasmid Characterization and Chromosome Analysis of Two netF...
PloS one2016; 11(2); e0148344; doi: 10.1371/journal.pone.0148344

Plasmid Characterization and Chromosome Analysis of Two netF+ Clostridium perfringens Isolates Associated with Foal and Canine Necrotizing Enteritis.

Abstract: The recent discovery of a novel beta-pore-forming toxin, NetF, which is strongly associated with canine and foal necrotizing enteritis should improve our understanding of the role of type A Clostridium perfringens associated disease in these animals. The current study presents the complete genome sequence of two netF-positive strains, JFP55 and JFP838, which were recovered from cases of foal necrotizing enteritis and canine hemorrhagic gastroenteritis, respectively. Genome sequencing was done using Single Molecule, Real-Time (SMRT) technology-PacBio and Illumina Hiseq2000. The JFP55 and JFP838 genomes include a single 3.34 Mb and 3.53 Mb chromosome, respectively, and both genomes include five circular plasmids. Plasmid annotation revealed that three plasmids were shared by the two newly sequenced genomes, including a NetF/NetE toxins-encoding tcp-conjugative plasmid, a CPE/CPB2 toxins-encoding tcp-conjugative plasmid and a putative bacteriocin-encoding plasmid. The putative beta-pore-forming toxin genes, netF, netE and netG, were located in unique pathogenicity loci on tcp-conjugative plasmids. The C. perfringens JFP55 chromosome carries 2,825 protein-coding genes whereas the chromosome of JFP838 contains 3,014 protein-encoding genes. Comparison of these two chromosomes with three available reference C. perfringens chromosome sequences identified 48 (~247 kb) and 81 (~430 kb) regions unique to JFP55 and JFP838, respectively. Some of these divergent genomic regions in both chromosomes are phage- and plasmid-related segments. Sixteen of these unique chromosomal regions (~69 kb) were shared between the two isolates. 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. perfringens strains. These results provide significant insight into the basis of canine and foal necrotizing enteritis and are the first to demonstrate that netF resides on a large and unique plasmid-encoded locus.
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Publication Date: 2016-02-09 PubMed ID: 26859667PubMed Central: PMC4747519DOI: 10.1371/journal.pone.0148344Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article discusses the fully sequenced genomes of two strains of bacteria (JFP55 and JFP838), which were found in animals suffering from foal and canine necrotizing enteritis. These strains contain a novel beta-pore-forming toxin, NetF, which appears to play a significant role in the disease.

Methodology and Genome Sequencing

  • The study utilized Single Molecule, Real-Time (SMRT) technology-PacBio and Illumina Hiseq2000 for genome sequencing.
  • It unveiled the complete genomes for two strains of Clostridium perfringens bacteria, JFP55 and JFP838, which had been previously linked to cases of necrotizing enteritis in foals and hemorrhagic gastroenteritis in canines respectively.
  • Each of the bacterial genomes comprised a single chromosome: the JFP55 strain with a 3.34 Mb chromosome, and the JFP838 strain with a 3.53 Mb chromosome.

Analysis of Plasmids

  • Five circular plasmids were identified in the genomes of the two strains.
  • Three of the five plasmids were found to be shared between the two sequenced genomes.
  • These included a plasmid encoding the NetF/NetE toxins, a plasmid encoding CPE/CPB2 toxins, and a putative bacteriocin-encoding plasmid.
  • The genes associated with pore-forming toxins (netF, netE, and netG) were located in unique pathogenicity zones on the tcp-conjugative plasmids.

Chromosomal Markers, Genes, and Comparative Genomes

  • The JFP55 chromosome held 2,825 protein-coding genes, while the JFP838 chromosome contained 3,014 protein-encoding genes.
  • Comparisons with existing C. perfringens chromosomes identified unique regions (48 totaling ~247kb and 81 totaling ~430kb) within the JFP55 and JFP838 chromosomes respectively.
  • Some distinctive chromosomal sections in both entities were related to bacteriophages and plasmids.

Findings and Implications

  • 16 unique sections (~69kb) overlapped between the two strains, which suggests these elements might have been acquired early in a clonal lineage of netF-positive C. perfringens strains.
  • Five of these shared sections formed a patchwork of plasmid-integrated segments.
  • The findings have brought important insights into the nature of canine and foal necrotizing enteritis. This is the first evidence of the netF gene being located within a large, unique plasmid-encoded locus.

Cite This Article

APA
Mehdizadeh Gohari I, Kropinski AM, Weese SJ, Parreira VR, Whitehead AE, Boerlin P, Prescott JF. (2016). Plasmid Characterization and Chromosome Analysis of Two netF+ Clostridium perfringens Isolates Associated with Foal and Canine Necrotizing Enteritis. PLoS One, 11(2), e0148344. https://doi.org/10.1371/journal.pone.0148344

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 11
Issue: 2
Pages: e0148344
PII: e0148344

Researcher Affiliations

Mehdizadeh Gohari, Iman
  • Department of Pathobiology, University of Guelph, Guelph, Ontario N1G 2W1, Canada.
Kropinski, Andrew M
  • Department of Pathobiology, University of Guelph, Guelph, Ontario N1G 2W1, Canada.
Weese, Scott J
  • Department of Pathobiology, University of Guelph, Guelph, Ontario N1G 2W1, Canada.
Parreira, Valeria R
  • Department of Pathobiology, University of Guelph, Guelph, Ontario N1G 2W1, Canada.
Whitehead, Ashley E
  • Department of Veterinary Clinical and Diagnostic Sciences, University of Calgary, Calgary, Alberta, CSB 101E, Canada.
Boerlin, Patrick
  • Department of Pathobiology, University of Guelph, Guelph, Ontario N1G 2W1, Canada.
Prescott, John F
  • Department of Pathobiology, University of Guelph, Guelph, Ontario N1G 2W1, Canada.

MeSH Terms

  • Animals
  • Bacterial Toxins / genetics
  • Chromosomes / genetics
  • Clostridium Infections / microbiology
  • Clostridium Infections / veterinary
  • Clostridium perfringens / genetics
  • Dogs / microbiology
  • Enteritis / microbiology
  • Enteritis / veterinary
  • Genetic Loci / genetics
  • Genomics
  • Horses / microbiology
  • Plasmids / genetics
  • Sequence Analysis
  • Species Specificity

Conflict of Interest Statement

The authors have declared that no competing interests exist.

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Citations

This article has been cited 25 times.
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