title: Detection of Clostridium botulinum neurotoxin genes (A-F) in dairy farms from Northern Germany using PCR: A case-control study.
abstract: Classical botulism in cattle mainly occurs after ingestion of feed contaminated with preformed toxin. In 2001 a form of botulism ("visceral botulism") was postulated to occur after ingestion of Clostridium (C.) botulinum cells or spores, followed by colonization of the intestine, and local production of botulinum neurotoxin (BoNT) causing chronic generalized disease. To verify the potential role of C. botulinum in the described syndrome, a case-control study was conducted, including 139 farms. Fecal samples, rumen content, water and silage samples were collected on each farm. Real time BoNT gene PCR assays were conducted after enrichment in RCM (Reinforced Clostridial Medium) at 37 °C and conventional PCRs after enrichment in MCM (Modified Cooked Meat Medium) at 30 °C. Furthermore, a direct detection of BoNT genes without prior enrichment was attempted. BoNT A, B, C, D, E and F genes were detected in animal samples from 25 (17.99%), 3 (2.16%), 0 (0.0%), 2 (1.44%), 1 (0.72%), and 3 (2.16%) farms, respectively. Eleven feed samples were positive for BoNT A gene. By enrichment a significant increase in sensitivity was achieved. Therefore, this should be an essential part of any protocol. No significant differences regarding BoNT gene occurrence could be observed between Case and Control farms or chronically diseased and clinically healthy animals within the particular category. Thus, the postulated form of chronic botulism in cows could not be confirmed. This study supports the general opinion that C. botulinum can occasionally be found in the rumen and intestine of cows without causing disease.
authors.affiliation: Institute of Food Quality and Food Safety, Research Center for Emerging Infections and Zoonoses, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, 30173 Hannover, Germany. Electronic address: svenja.fohler@tiho-hannover.de.
authors.forename: Svenja
authors.initials: S
authors.lastname: Fohler
authors.affiliation: Friedrich-Loeffler-Institut, Institute of Bacterial Infections and Zoonoses, Naumburger Straße 96a, 07743 Jena, Germany. Electronic address: sabrina_discher@yahoo.de.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Discher
authors.affiliation: Friedrich-Loeffler-Institut, Institute of Bacterial Infections and Zoonoses, Naumburger Straße 96a, 07743 Jena, Germany. Electronic address: eva.jordan@gmx.net.
authors.forename: Eva
authors.initials: E
authors.lastname: Jordan
authors.affiliation: Friedrich-Loeffler-Institut, Institute of Bacterial Infections and Zoonoses, Naumburger Straße 96a, 07743 Jena, Germany. Electronic address: Christian.Seyboldt@fli.bund.de.
authors.forename: Christian
authors.initials: C
authors.lastname: Seyboldt
authors.affiliation: Institute of Food Quality and Food Safety, Research Center for Emerging Infections and Zoonoses, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, 30173 Hannover, Germany. Electronic address: Guenter.Klein@tiho-hannover.de.
authors.forename: Guenter
authors.initials: G
authors.lastname: Klein
authors.affiliation: Friedrich-Loeffler-Institut, Institute of Bacterial Infections and Zoonoses, Naumburger Straße 96a, 07743 Jena, Germany. Electronic address: Heinrich.Neubauer@fli.bund.de.
authors.forename: Heinrich
authors.initials: H
authors.lastname: Neubauer
authors.affiliation: Clinic for Cattle, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, 30173 Hannover, Germany. Electronic address: martina.hoedemaker@tiho-hannover.de.
authors.forename: Martina
authors.initials: M
authors.lastname: Hoedemaker
authors.affiliation: Clinic for Cattle, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, 30173 Hannover, Germany. Electronic address: theresa.scheu@tiho-hannover.de.
authors.forename: Theresa
authors.initials: T
authors.lastname: Scheu
authors.affiliation: Department of Biometry, Epidemiology and Information Processing, University of Veterinary Medicine Hannover, Foundation, Buenteweg 2, 30559 Hannover, Germany. Electronic address: amely.campe@tiho-hannover.de.
authors.forename: Amely
authors.initials: A
authors.lastname: Campe
authors.affiliation: Department of Biometry, Epidemiology and Information Processing, University of Veterinary Medicine Hannover, Foundation, Buenteweg 2, 30559 Hannover, Germany. Electronic address: katharina.jensen@tiho-hannover.de.
authors.forename: Katharina
authors.initials: K
authors.lastname: Charlotte Jensen
authors.affiliation: Institute of Food Quality and Food Safety, Research Center for Emerging Infections and Zoonoses, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, 30173 Hannover, Germany. Electronic address: amir.abdulmawjood@tiho-hannover.de.
authors.forename: Amir
authors.initials: A
authors.lastname: Abdulmawjood
mesh_terms: D000818:Animals; D019274:Botulinum Toxins, Type A / Q000235:genetics / Q000302:isolation & purification*; D001906:Botulism / Q000382:microbiology / Q000662:veterinary*; D016022:Case-Control Studies; D002417:Cattle; D003014:Clostridium botulinum / Q000302:isolation & purification*; D003612:Dairying; D060766:Drinking Water / Q000737:chemistry; D000072480:Farms; D005243:Feces / Q000737:chemistry; D005260:Female; D005858:Germany; D016133:Polymerase Chain Reaction; D020033:Protein Isoforms / Q000235:genetics / Q000302:isolation & purification; D012417:Rumen / Q000737:chemistry; D012820:Silage / Q000032:analysis
chemical_list: D060766:Drinking Water; D020033:Protein Isoforms; D019274:Botulinum Toxins, Type A; C545476:incobotulinumtoxinA
keywords: Botulism; Chronic disease; Clostridium botulinum; Clostridium botulinum neurotoxin; Dairy herds; PCR