TY - JOUR

T1 - Detection of Clostridium botulinum neurotoxin genes (A-F) in dairy farms from Northern Germany using PCR

T2 - A case-control study

AU - Fohler, Svenja

AU - Discher, Sabrina

AU - Jordan, Eva

AU - Seyboldt, Christian

AU - Klein, Guenter

AU - Neubauer, Heinrich

AU - Hoedemaker, Martina

AU - Scheu, Theresa

AU - Campe, Amely

AU - Jensen, Katharina

AU - Abdulmawjood, Amir

N1 - Copyright © 2016 Elsevier Ltd. All rights reserved.

PY - 2016/6

Y1 - 2016/6

N2 - 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.

AB - 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.

KW - Animals

KW - Botulinum Toxins, Type A/genetics

KW - Botulism/microbiology

KW - Case-Control Studies

KW - Cattle

KW - Clostridium botulinum/isolation & purification

KW - Dairying

KW - Drinking Water/chemistry

KW - Farms

KW - Feces/chemistry

KW - Female

KW - Germany

KW - Polymerase Chain Reaction

KW - Protein Isoforms/genetics

KW - Rumen/chemistry

KW - Silage/analysis

U2 - 10.1016/j.anaerobe.2016.03.008

DO - 10.1016/j.anaerobe.2016.03.008

M3 - Journal article

C2 - 27016061

VL - 39

SP - 97

EP - 104

JO - Anaerobe

JF - Anaerobe

SN - 1075-9964

ER -