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

Mehdizadeh Gohari I, Parreira VR, Nowell VJ, Nicholson VM, Oliphant K, Prescott JF - PLoS ONE (2015)

Bottom Line: Mutation and complementation showed that only netF was associated with the cytotoxicity.Although netE and netG were not associated with cytotoxicity, immunoblotting with specific antisera showed these proteins to be expressed in vitro.The identifica-tion of this novel necrotizing toxin is an important advance in understanding the virulence of type A C. perfringens in specific enteric disease of animals.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathobiology, University of Guelph, Guelph, Ontario, Canada.

ABSTRACT
A role for type A Clostridium perfringens in acute hemorrhagic and necrotizing gastroenteritis in dogs and in necrotizing enterocolitis of neonatal foals has long been suspected but incompletely characterized. The supernatants of an isolate made from a dog and from a foal that died from these diseases were both found to be highly cytotoxic for an equine ovarian (EO) cell line. Partial genome sequencing of the canine isolate revealed three novel putative toxin genes encoding proteins related to the pore-forming Leukocidin/Hemolysin Superfamily; these were designated netE, netF, and netG. netE and netF were located on one large conjugative plasmid, and netG was located with a cpe enterotoxin gene on a second large conjugative plasmid. Mutation and complementation showed that only netF was associated with the cytotoxicity. Although netE and netG were not associated with cytotoxicity, immunoblotting with specific antisera showed these proteins to be expressed in vitro. There was a highly significant association between the presence of netF with type A strains isolated from cases of canine acute hemorrhagic gastroenteritis and foal necrotizing enterocolitis. netE and netF were found in all cytotoxic isolates, as was cpe, but netG was less consistently present. Pulsed-field gel electrophoresis showed that netF-positive isolates belonged to a clonal population; some canine and equine netF-positive isolates were genetically indistinguishable. Equine antisera to recombinant Net proteins showed that only antiserum to rNetF had high supernatant cytotoxin neutralizing activity. The identifica-tion of this novel necrotizing toxin is an important advance in understanding the virulence of type A C. perfringens in specific enteric disease of animals.

No MeSH data available.


Related in: MedlinePlus

Dendogram of Clostridium perfringens isolates.Dendogram of C.perfringens isolates typed by pulsed-field gel electrophoresis and analysed using BioNumerics software. The BioNumerics software used was version 7.1 from Applied Maths, Austin, TX.
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pone.0122684.g005: Dendogram of Clostridium perfringens isolates.Dendogram of C.perfringens isolates typed by pulsed-field gel electrophoresis and analysed using BioNumerics software. The BioNumerics software used was version 7.1 from Applied Maths, Austin, TX.

Mentions: There is increasing evidence that C. perfringens associated with different enteric diseases may belong to clonal populations. We used PFGE to examine the relatedness of canine and equine netF+ and netF- isolates from our strain collection. The PFGE analysis and the dendogram are shown in Fig 5. Out of the 70 isolates (35 equine and 35 canine) typed by PFGE, 56 (80%) individual pulse types were identified based on an identical banding pattern (genetic similarity of 100%). Eighteen of these 56 were positive for netF, 37 were negative for netF, and one type contained both netF+ and netF- isolates that were equivalent. There was no pattern distinguishable between equine and canine types (Fig 5).


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

Mehdizadeh Gohari I, Parreira VR, Nowell VJ, Nicholson VM, Oliphant K, Prescott JF - PLoS ONE (2015)

Dendogram of Clostridium perfringens isolates.Dendogram of C.perfringens isolates typed by pulsed-field gel electrophoresis and analysed using BioNumerics software. The BioNumerics software used was version 7.1 from Applied Maths, Austin, TX.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4390311&req=5

pone.0122684.g005: Dendogram of Clostridium perfringens isolates.Dendogram of C.perfringens isolates typed by pulsed-field gel electrophoresis and analysed using BioNumerics software. The BioNumerics software used was version 7.1 from Applied Maths, Austin, TX.
Mentions: There is increasing evidence that C. perfringens associated with different enteric diseases may belong to clonal populations. We used PFGE to examine the relatedness of canine and equine netF+ and netF- isolates from our strain collection. The PFGE analysis and the dendogram are shown in Fig 5. Out of the 70 isolates (35 equine and 35 canine) typed by PFGE, 56 (80%) individual pulse types were identified based on an identical banding pattern (genetic similarity of 100%). Eighteen of these 56 were positive for netF, 37 were negative for netF, and one type contained both netF+ and netF- isolates that were equivalent. There was no pattern distinguishable between equine and canine types (Fig 5).

Bottom Line: Mutation and complementation showed that only netF was associated with the cytotoxicity.Although netE and netG were not associated with cytotoxicity, immunoblotting with specific antisera showed these proteins to be expressed in vitro.The identifica-tion of this novel necrotizing toxin is an important advance in understanding the virulence of type A C. perfringens in specific enteric disease of animals.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathobiology, University of Guelph, Guelph, Ontario, Canada.

ABSTRACT
A role for type A Clostridium perfringens in acute hemorrhagic and necrotizing gastroenteritis in dogs and in necrotizing enterocolitis of neonatal foals has long been suspected but incompletely characterized. The supernatants of an isolate made from a dog and from a foal that died from these diseases were both found to be highly cytotoxic for an equine ovarian (EO) cell line. Partial genome sequencing of the canine isolate revealed three novel putative toxin genes encoding proteins related to the pore-forming Leukocidin/Hemolysin Superfamily; these were designated netE, netF, and netG. netE and netF were located on one large conjugative plasmid, and netG was located with a cpe enterotoxin gene on a second large conjugative plasmid. Mutation and complementation showed that only netF was associated with the cytotoxicity. Although netE and netG were not associated with cytotoxicity, immunoblotting with specific antisera showed these proteins to be expressed in vitro. There was a highly significant association between the presence of netF with type A strains isolated from cases of canine acute hemorrhagic gastroenteritis and foal necrotizing enterocolitis. netE and netF were found in all cytotoxic isolates, as was cpe, but netG was less consistently present. Pulsed-field gel electrophoresis showed that netF-positive isolates belonged to a clonal population; some canine and equine netF-positive isolates were genetically indistinguishable. Equine antisera to recombinant Net proteins showed that only antiserum to rNetF had high supernatant cytotoxin neutralizing activity. The identifica-tion of this novel necrotizing toxin is an important advance in understanding the virulence of type A C. perfringens in specific enteric disease of animals.

No MeSH data available.


Related in: MedlinePlus