ABSTRACT
Botulism in cattle is a rare but serious disease. In Germany there is no obligation to report botulism in animals and therefore a precise morbidity rate is not available. In this manuscript we describe an outbreak of Clostridium (C.) botulinum neurotoxin (BoNT) intoxication in a Saxony-Anhalt dairy cow stock of 286 Holstein-Friesian cows and offspring in spring/summer 2009 and its diagnostic approach. 122 animals showed clinical signs of BoNT intoxication. 115 of the affected animals (40.2% of the herd) independent of age died or had to be euthanized. Therapeutic attempts failed in almost all diseased cows, only four calves and three heifers recovered. Diagnostic samples of several animals (n = 4) (liver, ruminal and intestinal contents) and feed (n = 6) were tested for BoNT genes by polymerase chain reaction (PCR). BoNT gene type D was found in several (n = 8) organ samples. The PCR results allowed a preselection of samples for BoNT that were then tested by the mouse bioassay. Thus, the number of mice being inoculated in the mouse bioassay could be reduced. The mouse bioassay turned out positive (wasp-waist) in three preselected organ samples and the neutralization test of one sample with type-specific antitoxin confirmed the presence of BoNT type D. We succeeded in isolating a C. botulinum strain from a liver sample which was typed as a D/C mosaic strain by sequence analysis of the toxin gene. However, the source of the BoNT intoxication could not be traced back.
Subject(s)
Botulism/veterinary , Cattle Diseases/microbiology , Clostridium botulinum type D/classification , Disease Outbreaks/veterinary , Animals , Botulism/diagnosis , Botulism/epidemiology , Botulism/microbiology , Cattle , Cattle Diseases/diagnosis , Cattle Diseases/epidemiology , Clostridium botulinum type D/genetics , Clostridium botulinum type D/isolation & purification , Clostridium botulinum type D/metabolism , Dairying , Female , Germany/epidemiology , Mice , Tetanus Toxin/genetics , Tetanus Toxin/metabolismABSTRACT
Pasteurized milk, liquid egg, minced meat, and various salads were artificially contaminated with varying numbers of cells from six Escherichia coli ( E. coli ) strains able to produce heat-stable enterotoxins (ST). The ST-producing E. coli were detected by the following procedure within 24 h without isolation by cultivation. After enrichment of the food sample in GN broth (4 h at 37°C), the material was transferred to brain heart infusion broth, incubated (16-18 h at 37°C), centrifuged (20 min, 7000 g) and heated to 80°C for 10 min, the supernatant was tested with the infant mouse test. The sensitivity (= ratio of detectable E. coli per total microbial numbers in the food sample) of the test procedure was high even in many food samples with a considerable competitive microbial flora. The procedure was used to test 419 routine food samples. Enterotoxigenic bacteria were found in 7 samples of liquid egg and 4 samples of salad. The test is recommended as a rapid screening test in food control.
