title: Confirmation of botulism in birds and cattle by the mouse bioassay and Endopep-MS.
abstract: There have been several outbreaks of botulism among poultry and wild birds in Sweden in recent years. The National Veterinary Institute of Sweden (SVA) has identified botulinum neurotoxin (BoNT)/C1 or the mosaic BoNT/C1D using the mouse bioassay. This is believed to be the first report on the application of the Endopep mass spectrometry (Endopep-MS) method to selected clinical animal (serum and liver) samples and a feed sample that had previously given positive test results with the mouse bioassay. In the mouse bioassay eight of the eleven samples were found to be neutralized by both BoNT/C1 and /D antitoxins; the other three were neutralized only by BoNT/C1 antitoxin, but the mice showed a prolonged survival time when the samples had been treated with /D antitoxin. The Endopep-MS analysis, on the other hand, demonstrated only BoNT/C1 activity for all eleven samples. This suggests that at least eight of the samples were of the chimeric toxin type BoNT/C1D, where the enzymically active site is identical to that of BoNT/C1, while other parts of the protein contain sequences of BoNT/D. This is the first step of a cross-validation between the established mouse bioassay and the Endopep-MS of serotypes BoNT/C1 and /C1D. Endopep-MS is concluded to have potential as an attractive alternative to the mouse bioassay.
authors.affiliation: Uppsala University, Division of Analytical Pharmaceutical Chemistry, Biomedical Centre, PO Box 574, SE-751 23 Uppsala, Sweden.|National Veterinary Institute (SVA), Department of Chemistry, Environment and Feed Hygiene, SE-751 89 Uppsala, Sweden.
authors.forename: Mikael
authors.initials: M
authors.lastname: Hedeland
authors.affiliation: Centers for Disease Control and Prevention, NCEH/DLS, 4770 Buford Hwy NE, Atlanta, GA 30341-3724, USA.
authors.forename: Hercules
authors.initials: H
authors.lastname: Moura
authors.affiliation: National Veterinary Institute (SVA), Department of Bacteriology, SE-751 89 Uppsala, Sweden.
authors.forename: Viveca
authors.initials: V
authors.lastname: Båverud
authors.affiliation: Centers for Disease Control and Prevention, NCEH/DLS, 4770 Buford Hwy NE, Atlanta, GA 30341-3724, USA.
authors.forename: Adrian R
authors.initials: AR
authors.lastname: Woolfitt
authors.affiliation: Uppsala University, Division of Analytical Pharmaceutical Chemistry, Biomedical Centre, PO Box 574, SE-751 23 Uppsala, Sweden.|National Veterinary Institute (SVA), Department of Chemistry, Environment and Feed Hygiene, SE-751 89 Uppsala, Sweden.
authors.forename: Ulf
authors.initials: U
authors.lastname: Bondesson
authors.affiliation: Centers for Disease Control and Prevention, NCEH/DLS, 4770 Buford Hwy NE, Atlanta, GA 30341-3724, USA.
authors.forename: John R
authors.initials: JR
authors.lastname: Barr
mesh_terms: D032761:Animal Experimentation; D000818:Animals; D001681:Biological Assay / Q000379:methods*; D001715:Bird Diseases / Q000175:diagnosis*; D001717:Birds; D001906:Botulism / Q000175:diagnosis / Q000662:veterinary*; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis*; D008099:Liver / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D009500:Neutralization Tests; D044967:Serum / Q000737:chemistry; D013548:Sweden