A major outbreak of botulism in cattle being fed ensiled poultry litter

Eighty of a group of 150 housed beef cattle showed classical signs of botulism after eating a batch of ensiled poultry litter. Sixty-eight of the animals died and Clostridium botulinum type C toxin was detected in 18 of 22 sera examined. C botulinum organisms were isolated from the ensiled litter and type C toxin was demonstrated in samples of decomposed poultry carcases present in the litter. This outbreak of bovine botulism was the most serious to have been recorded in Europe and was the first associated with feeding ensiled poultry litter.     

Type C botulism in cattle being fed ensiled poultry litter

A botulinum toxin from ensiled poultry litter which caused a major outbreak of bovine botulism was characterised as type C1. The litter produced transient ataxia when fed to two experimental calves and the clinical signs were accompanied by a transient appearance of serum toxin. Type C1 toxin was demonstrated in muscle tissues which had been taken during the outbreak from an affected animal with high circulating serum toxin, and held frozen for seven months. Clostridium botulinum type C organisms were demonstrated in faeces from another affected animal and also in kidney tissue from a third animal. These observations have implications for the diagnosis and management of future outbreaks of botulism and for the potential health risk from the meat of affected animals.     

Outbreak of botulism in a dairy herd in Turkey

In this study, the clinical findings and results of haematological and biochemical analyses of 26 cattle with botulism were evaluated. The most important clinical signs in the affected cattle included: decreased appetite, ataxia, difficulty to rise, loss of tongue tone, salivation and bradycardia. A definitive diagnosis of botulism was based on demonstration of the preformed toxin in ruminal and intestinal contents and feed materials including poultry litter, by mouse inoculation test. This study is the first confirmation, by direct toxin isolation, of Clostridium botulinum type C and Clostridium botulinum type D in cattle, in Turkey.     

Use of enzyme-linked immunoassays for antibody to types C and D botulinum toxins for investigations of botulism in cattle

The development of specific enzyme-linked immunosorbent assays (ELISA) for antibody to types C and D Clostridium botulinum toxins for investigation of botulism in cattle is described. Partially purified type C and D toxins were used as antigens to develop these ELISAs. Specificity of the ELISAs was evaluated on sera from 333 adult beef and dairy cattle from areas with no history or evidence of botulism in animals or water birds. The test was also evaluated on sera from 41 herds that included herds vaccinated against botulism, confirmed botulism cases and herds from areas where the disease is considered endemic. The ELISAs detected the presence of antibody to botulinum toxins in samples from vaccinated cattle and both convalescent and clinically normal animals from unvaccinated herds with outbreaks of botulism. Antibody was also found in unvaccinated animals from herds in which there had been no diagnosed botulism cases in areas where botulism was considered endemic. Sera from some unvaccinated cattle with high ELISA reactivity was shown to be protective for mice in botulinum toxin neutralisation tests. The use of these tests in investigations of botulism in cattle is discussed.     

An outbreak of type C botulism in broiler chickens.

An outbreak of type C botulism involving three-week-old broiler chickens on deep litter is described. No direct source of toxin was found. Cl botulinum type C was distributed widely in the litter, and several thousand per gram were demonstrated both in the litter and in the intestinal contents of chicken.     

Presumptive diagnosis of Clostridium botulinum type D intoxication in a herd of feedlot cattle

Fifty-two feedlot cattle exhibited clinical signs suggestive of botulism. Clostridium botulinum type D organisms were recovered from ruminal fluid of 4 of the 5 affected animals tested and were isolated from bakery waste fed to the cattle. Clostridium botulinum type D has not been reported previously in Canadian cattle.     

Suspected botulism in cattle associated with poultry litter

Botulism was diagnosed clinically in grazing cattle on three closely sited dairy farms. The evidence suggests that the source of the toxin was poultry carcases containing types C and D Clostridium botulinum. These organism were also found in the alimentary tract of affected animals. Silage is suspected as having acted as a vehicle for the organisms and, or, toxins in cases which occurred later in housed cattle on one of the farms.     

Surveillance of suspect animal toxicoses with potential food safety implications in England and Wales between 1990 and 2002

The potential chemical contamination incidents investigated by the Veterinary Laboratories Agency (VLA) between 1990 and 2002 are reviewed. Incidents were identified in the course of the VLA's surveillance of food animal disease and further investigations were carried out on behalf of the Food Standards Agency in order to identify and control the contamination of food animals and animal products with chemical hazards. In total 876 incidents were investigated, of which 588 involved the poisoning of cattle with heavy metals. There were 63 incidents involving the exposure of cattle, sheep, pigs and poultry to the accidental misuse of veterinary medicines, pesticides or rodenticides, and 50 incidents involving their exposure to microbiological toxins, particularly botulism.     

Multiplex real-time PCR SYBR Green for detection and typing of group III Clostridium botulinum

Clostridium botulinum type C and type D belonging to the group III organisms, are mainly responsible for animal botulism outbreaks. Clinical signs alone are often insufficient to make a diagnosis of botulism and a laboratory confirmation is required. Laboratory confirmation can be performed by demonstrating the presence of botulinum neurotoxins in serum, gastrointestinal contents, liver, wound of sick or dead animals, or by demonstrating the presence of C. botulinum in gastrointestinal contents, liver, and wound. Demonstration of spores in gastrointestinal contents or tissue of animals with clinical signs indicative of botulism reinforces the clinical diagnosis. With the aim of detecting and typing C. botulinum group III organisms, a multiplex real-time PCR SYBR Green was developed and in-house validated. Selectivity, limit of detection, relative accuracy, relative specificity, relative sensitivity, and repeatability of the method were investigated. The multiplex real-time PCR SYBR green used showed a 100% selectivity, 100% relative accuracy, 100% relative specificity, 100% relative sensitivity and a limit of detection of 277 and 580 DNA copies for C. botulinum type C and C. botulinum type D, respectively. The method reported here represents a suitable tool for laboratory diagnosis of type C and D botulism and for testing a large number of samples collected during the animal botulism surveillance and prevention activities.     

Botulism in chickens associated with elevated iron levels

Clostridium botulinum type C toxicosis was diagnosed by the mouse inoculation test in two outbreaks of botulism in commercial broiler and roaster chickens. One case involved 7-wk-old commercial roaster chickens, and the other involved 15-day-old commercial broiler chickens. A definitive point source for preformed C. botulinum exotoxin was not identified in either case investigation. Elevated iron concentrations in the drinking water and/or feed may have presented a significant risk factor that may have resulted in intestinal proliferation of C. botulinum and subsequent botulism.     

Avian botulism and the high prevalence of Clostridium botulinum in the Norflok Broads.

An account is given of a severe outbreak of type C botulism in waterfowl that occurred on the Norfolk Broads during the exceptionally warm summer of 1975. Forty-five mud samples were collected from 22 well distributed aquatic sites representing a considerable proportion of the total number of Broads. All samples except one (ie, 97-8 per cent) were shown to contain Clostridium botulinum and 58 per cent contained more than one type of the organism. Types B, C and E were demonstrated in 62-2 per cent, 51-1 per cent and 60 per cent of samples respectively. Recent surveys, made by identical methods, of aquatic environments in the London area and the Camargue (France) showed prevalences of Cl botulinum of 72-5 per cent and 4-5 per cent respectively. It seems likely that the Norfolk Broads will continue to present a risk to waterfowl from botulism in future hot summers.     

Determination of the median toxic dose of type C botulinum toxin in lactating dairy cows.

Because of the difficulty in identifying botulinum toxin in cattle, it is hypothesized that cattle are sensitive to levels of toxin below the detection limits of current diagnostic techniques (the mouse protection bioassay and the immunostick enzyme-linked immunosorbent assay [ELISA] for type C botulinum toxin). Using an up-down method for toxicologic testing, the median toxic dose (MTD50) for cattle was determined. Four lactating Holstein cows were dosed at 0.125 or 0.25 ng/kg with Clostridium botulinum type C toxin and failed to develop clinical signs of botulism during the 7-day observation period. Three cows given 0.50 ng/kg of toxin developed clinical signs of botulism. From these results, the MTD50 was calculated at 0.388 ng/kg (3.88 mouse lethal doses/kg) using the trim-logit method. These results suggest that cattle are 12.88 times more sensitive to type C botulinum toxin than a mouse on a per kilogram weight basis. The mouse protection bioassay and the immunostick ELISA for type C botulinum toxin failed to identify the presence of the toxin in the serum, blood, and milk samples taken from all 7 animals.     

Investigation of serology for diagnosis of outbreaks of botulism in cattle

Serology has been used to diagnose retrospectively types C and D outbreaks of botulism in cattle in Australia and this study has investigated whether the approach would be applicable in England and Wales. Three hundred sera from routine surveillance submissions in England and Wales were used as a negative control population. Some stored sera were available from a small number of clinical cases of botulism and 125 samples were collected from cohort groups of clinical cases in four new outbreaks of botulism. Three of these outbreaks were identified as being caused by type D Clostridium botulinum toxin. Sera were tested by antibody ELISA in laboratories in Australia and Germany. There was no increase in the proportion of animals seropositive to type C or D antibody in the botulism-associated cattle. The proportion of samples which were seropositive to type D antibodies was <2% in both the negative control and outbreak populations. It was concluded that single time serology is unlikely to be helpful for retrospective diagnosis of outbreaks of type D botulism in England and Wales.     

Susceptibility of Foxes to Clostridium Botulinum Type C and E Toxins

Investigations were performed to determine the exact susceptibility of foxes to Clostridium botulinum type C and E toxins.Doses of 5 mill. MLD type C toxin mixed with the feed did not cause symptoms of botulism in either cubs or adult foxes. Subcutaneous injections of 300,100(0 MLD or more were fatal to cubs, while 750,000 MLD caused the death of all adults.Regarding type E toxin, doses of 1 mill. MLD affected neither cubs nor adults on oral administration. Subcutaneously injected doses of 5,000 MLD or more killed all cubs, while 10,000 MLD was required to produce lethal effect on adult animals.The conclusion made is that foxes are highly resistant to both type C and E Clostridium botulinum toxins following oral application. It is further revealed that foxes are 60–70 times more susceptible to type E than to type C toxin when injected subcutaneously.     

Diagnosis and treatment of botulism in lions

Six circus lions (Panthera leo) showed neurological and gastrointestinal signs after consuming casualty broiler chickens. Signs included ataxia, hindlimb paralysis and recumbency. Neurological examination of two affected males showed paralysis of extraocular muscles, fixed dilated pupils and inability to swallow. Replacement fluids and antibiotics were given and Clostridium botulinum type C antitoxin was found in serum samples. Type C antitoxin was not then available and therapy was started in one lioness with guanidine hydrochloride. Convulsions were controlled by diazepam but this animal died. One of the two males was given type C antitoxin; both were given anabolic steroids. All the remaining animals made slow recoveries over varying periods; one lion was recumbent for 41 days. No lion developed respiratory paralysis; other animals which had consumed the chickens remained healthy. Aspects of the treatment of botulism in animals are discussed.     

Toxicoinfectious botulism in commercial caponized chickens

During the summer of 2003, two flocks of commercial broiler chickens experienced unusually high death losses following caponizing at 3 wk of age and again between 8 and 14 wk of age. In September, fifteen 11-wk-old live capons were submitted to the Iowa State University Veterinary Diagnostic Laboratory for assistance. In both flocks, the second episode of elevated mortality was associated with incoordination, flaccid paralysis of leg, wing, and neck muscles, a recumbent body posture characterized by neck extension, and diarrhea. No macroscopic or microscopic lesions were detected in affected chickens. Hearts containing dotted blood and ceca were submitted to the National Wildlife Health Center in Madison, WI. Type C. botulinum toxin was identified in heart blood and ceca by mouse bioassay tests. Enzyme-linked immunosorbent assay tests on heart blood samples were also positive for type C. botulinum toxin. Clostridium botulinum was isolated from the ceca and genes encoding type C. botulinum toxin were detected in cecal contents by a polymerase chain reaction test. Chickens are less susceptible to botulism as they age, and this disease has not previously been documented in broilers as old as 14 wk of age. Wound contamination by spores of C. botulinum may have contributed to the unusually high death losses following caponizing.     

Susceptibility of mink to Clostridium botulinum type E toxin

Experiments aiming at elucidation of the toxicity of Clostridium botulinum type E for mink are described. The observations indicate that amounts in the order of 2 x10^s intraperitoneal MLD (mice) or approximately 200 MLD per g of type E toxin will kill a mink after oral administration. The symptoms observed in the animals were atypical as there was an unusually short period between administration of the toxin and the onset of symptoms and deaths of the animals. Similar results were obtained when Clostridium botulinum type E toxin was fed to Swiss mice. When mice were protected by subcutaneous injections of type E antitoxin prior to feeding the animals survived without showing any symptoms.Subcutaneous injection of type E toxin in amounts of the order of 2 x10^s intraperitoneal MLD (mice) killed mink, and typical symptoms of botulism were observed. This quantity corresponds to ap-proximately 2 intraperitoneal MLD (mice) per g.Comparison is made with previous observations obtained in similar experiments made with Clostridium botulinum type C toxin. It is shown that mink arc substantially less susceptible to type E than to type C toxin when the toxins are given by mouth. On this basis previous results in reports on outbreaks of botulism in mink caused by Clostridium botulinum type E may be regarded as questionable.     

Cardiomyopathy of ruminants induced by the litter of poultry fed on rations containing the ionophore antibiotic, maduramicin. I. Epidemiology, clinical signs and clinical pathology.

The epidemiological, clinical and clinical pathological findings in 20 cattle and 4 sheep from 15 outbreaks of poultry litter toxicity in South Africa over the past 6 years are documented. In 6 outbreaks, the litter emanated from batteries where maduramicin had been incorporated into rations of broilers. According to circumstantial evidence the litter involved in the 9 other outbreaks was also derived from broilers which had been fed on rations containing an ionophore. The litter was fed ad libitum to the affected stock or constituted 30-80% by volume of their rations. The principal sign manifested was sudden mortality of up to 70% of the herd or flock, usually within 20-40 days of commencement of feeding of poultry litter. A few cattle developed signs of congestive heart failure, and stiffness was commonly seen in sheep. In a dosing trial with poultry litter involving 1 steer and 6 sheep, the steer and a sheep died suddenly and a second sheep was destroyed in extremis. Tachycardia and/or cardiac arrythmia were recorded in 5 sheep, and the activity of aspartate transaminase (AST) and/or lactate dehydrogenase (LD) in the sera of 4 was elevated. Since the cardiac lesions in field cases were similar to those of ionophore poisoning and broiler rations containing maduramicin was a common factor in several outbreaks, toxic litter from some of these outbreaks were tested for the presence of this compound. Analysis by high performance liquid chromatography of litter from 2 specimens of outbreaks revealed that they contained 2.5 ppm and 6.1 ppm maduramicin.(ABSTRACT TRUNCATED AT 250 WORDS)     

Prevalence of wet litter and the associated risk factors in broiler flocks in the United Kingdom

A postal questionnaire was sent to the managers of 857 broiler farms in the UK to determine the prevalence and risk factors for wet litter. The response rate was 75 per cent. Wet litter was reported by 75 per cent (95 per cent confidence interval [CI] 71.3 to 78.3) of the respondents in at least one flock during the year 2001 and 56.1 per cent (95 per cent CI 52.0 to 60.0) of them reported that they had an outbreak of wet litter in their most recently reared flock. Wet litter occurred more often during the winter months and farms using side ventilation systems were at an increased risk (odds ratio 1.74; 95 per cent CI 1.09 to 2.76). A multivariable analysis was carried out using two different definitions of wet litter as outcome variables - all cases of wet litter, and cases of wet litter associated with disease. Consistent risk factors for both outcomes were coccidiosis, feed equipment failures and the availability of separate farm clothing for each house. Cases of wet litter associated with disease were reported by 33.7 per cent (95 per cent CI 28.8 to 39.1) of the managers in their last flock and were associated with the use of hand sanitisers and broiler houses with walls made of concrete.