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PubMed2003

Two cases of equine grass sickness with evidence for soil-borne origin involving botulinum neurotoxin.

Botulism is caused by different types of Clostridium botulinum, a soil bacterium. Equine grass sickness (equine dysautonomia) is suspected of being a clinical form of this disease. On a stud where this disease occurred twice within 8 months, grass and soil samples and necropsy specimens of one horse were tested for the presence of bacterial forms and toxin of C. botulinum. Different types and type mixtures (A-E) of C. botulinum and botulinum neurotoxin were found. For the first time, it has been shown that green grass blades contain botulinum toxin. The results support the hypothesis that equine grass sickness is a clinical form of botulism, a soil-borne disease.

Animals↗
PubMed2000

A taste of honey.

Infantile botulism classically presents with a triad of clinical features: Bulbar palsies (slow/absent pupil response) Alert Absent fever Other common features are: Constipation, ptosis and poor feeding The diagnosis is a clinical one, confirmed by EMG and by testing stool for the organism, C. botulinum, or its toxin. Parents should be advised not to dip pacifiers in honey. In future Botulinum Immune Globulin may be available and should be considered if further trials are published showing benefit. Nosocomial infections, such as RSV, are important and should be carefully avoided, particularly in patients recovering from botulism.

Botulism↗
PubMed2005

Clostridium botulinum: a bug with beauty and weapon.

Clostridium botulinum, a Gram-positive, anaerobic spore-forming bacteria, is distinguished by its significant clinical applications as well as its potential to be used as bioterror agent. Growing cells secrete botulinum neurotoxin (BoNT), the most poisonous of all known poisons. While BoNT is the causative agent of deadly neuroparalytic botulism, it also serves as a remarkably effective treatment for involuntary muscle disorders such as blepharospasm, strabismus, hemifacial spasm, certain types of spasticity in children, and other ailments. BoNT is also used in cosmetology for the treatment of glabellar lines, and is well-known as the active component of the anti-aging medications Botox and Dysport. In addition, recent reports show that botulinum neurotoxin can be used as a tool for pharmaceutical drug delivery. However, BoNT remains the deadliest of all toxins, and is viewed by biodefense researchers as a possible agent of bioterrorism (BT). Among seven serotypes, C. botulinum type A is responsible for the highest mortality rate in botulism, and thus has the greatest potential to act as biological weapon. Genome sequencing of C. botulinum type A Hall strain (ATCC 3502) is now complete, and has shown the genome size to be 3.89 Mb with a G+C content of approximately 28.2%. The bacterium harbors a 16.3 kb plasmid with a 26.8% G+C content--slightly lower than that of the chromosome. Most of the virulence factors in C. botulinum are chromosomally encoded; bioinformatic analysis of the genome sequence has shown that the plasmid does not harbor toxin genes or genes for related virulence factors. Interestingly, the plasmid does harbor genes essential to replication, including dnaE, which encodes the alpha subunit of DNA polymerase III which has close similarity with its counterpart in C. perfringens strain 13. The plasmid also contains similar genes to those that encode the ABC-type multidrug transport ATPase, and permease. The presence of ABC-type multidrug transport ATPase, and permease suggests putative involvement of efflux pumps in bacteriocin production, modification, and export in C. botulinum. The C. botulinum plasmid additionally harbors genes for LambdaBa04 prophage and site-specific recombinase that are similar to those found in the Ames strain of Bacillus anthracis; these genes and their products may play a role in genomic rearrangement. Completion of genome sequencing for C. botulinum will provide an opportunity to design genomic and proteomic-based systems for detecting different serotypes of C. botulinum strains in the environment. The completed sequence may also facilitate identification of potential virulence factors and drug targets, as well as help characterize neurotoxin-complexing proteins, their polycistronic expression, and phylogenetic relationships between different serotypes.

Antibodies, Bacterial↗
PubMed1982

Resistance of mice with limited intestinal flora to enteric colonization by Clostridium botulinum.

Infant botulism is an age-dependent illness that is caused by the toxin produced by Clostridium botulinum infecting the intestinal tract. Because of composition of the intestinal microflora determines the resistance of mice to enteric colonization by C. botulinum, attempts were made to identify the kinds of bacteria that prevent this in vivo growth. Orogastric challenges of 50 spores of C. botulinum type A were given to adult germfree mice, which are highly susceptible, and to gnotobiotic adult mice carrying the eight species comprising the Charles River Altered Schaedler flora or two or three of the limited number of species constituting a different flora (the University of Wisconsin Gnotobiote Laboratory [UW-GL] flora). These floras did not prevent infection due to C. botulinum; however, death rates among the mice with defined floras were significantly lower than those among germfree mice exposed to C. botulinum. Botulinum toxin continued to be produced while animals surviving nearly lethal cases of botulism convalesced slowly but uneventfully. Gnotobiotic mice with the complete UW-GL flora were not infected when challenged with 10(5) spores of C. botulinum.

Animals↗
PubMed1999

Isolation and characterisation of neurotoxigenic Clostridium butyricum from soil in China.

Soil specimens collected from a site around the home of patients with food-borne type E. botulism probably caused by neurotoxigenic Clostridium butyricum in Guanyun, Jiangsu province, China, were examined for the presence of neurotoxigenic C. butyricum. Five lakeside sites of Weishan lake, in an area near to the sites where the type E. botulism outbreaks caused by neurotoxigenic C. butyricum occurred were also surveyed. Type E toxin-producing C. butyricum was isolated from soil from four sites including the site in Guanyun. Polymerase chain reaction assay demonstrated the presence of the type E toxin gene in all the toxigenic isolates. The biochemical properties of the isolates from the Guanyun soil and the lakeside soil were identical except for inulin fermentation and starch hydrolysis properties. These results indicate that neurotoxigenic C. butyricum has its principal habitat in soil.

Bacterial Toxins↗
PubMed1991

Characterization of neurotoxigenic Clostridium butyricum strain by DNA hybridization test and by in vivo and in vitro germination tests of spores.

The germination of spores of a neurotoxigenic Clostridium butyricum strain (BL 6340), which was isolated from infant botulism in Italy, and that of a nontoxigenic C. butyricum type strain (NCIB 7423) were studied. The spores of BL 6340 strain were killed at 80 C for 10 min, and required the mixture of L-alanine, L-lactate, glucose and bicarbonate for their optimal germination. These characteristics are the same as those of Clostridium botulinum type E strain, but different from those of NCIB 7423 strain. In a hybridization test, however, the labeled DNAs extracted from NCIB 7423 strain highly (98%) hybridized to the DNAs of the BL 6340 strain, but little (45%) to the DNAs of C. botulinum type E strain. The biochemical properties of the BL 6340 and NCIB 7423 strains were identical, but different from those of C. botulinum type E. These data confirmed that the BL 6340 strain belongs to C. butyricum species, but that only its characteristics of toxin production, its minimum requirements for germination, and the behavior of its spores to heat treatment are the same as those of C. botulinum type E. When conventionally raised suckling mice were injected with 5 x 10(7) spores of BL 6340 strain intra- or orogastrically, botulism was not observed. However, 8- to 13-day-old mice had type E botulinum toxin in the large intestine 3 days after introduction of its spores.

Animals↗
PubMed2006

Application of PCR for detection of Clostridium botulinum type D in bovine samples.

Diagnosis of botulism in cows is obtained by detecting the neurotoxin and/or Clostridium botulinum in the suspected animal. The standard method for detecting the toxin is the mouse bioassay. However, in recent years, the use of mice has become very costly and inconvenient in some facilities, and public pressure has been increasing to find alternatives to live animal bioassays. In this manuscript, we describe the use of the polymerase chain reaction (PCR) procedures in the diagnosis field cases of bovine type D botulism. Bovine samples from clinical cases diagnosed as C. botulinum type D according by clinical symptoms and bioassay resulted in expected PCR product ( approximately 497 bp) similar to the C. botulinum type D NCTC 8265 strain while the gene product was confirmed by sequence data.

Animals↗
PubMed1999

A predictive model that describes the effect of prolonged heating at 70 to 90 degrees C and subsequent incubation at refrigeration temperatures on growth from spores and toxigenesis by nonproteolytic Clostridium botulinum in the presence of lysozyme.

Refrigerated processed foods of extended durability such as cook-chill and sous-vide foods rely on a minimal heat treatment at 70 to 95 degrees C and then storage at a refrigeration temperature for safety and preservation. These foods are not sterile and are intended to have an extended shelf life, often up to 42 days. The principal microbiological hazard in foods of this type is growth of and toxin production by nonproteolytic Clostridium botulinum. Lysozyme has been shown to increase the measured heat resistance of nonproteolytic C. botulinum spores. However, the heat treatment guidelines for prevention of risk of botulism in these products have not taken into consideration the effect of lysozyme, which can be present in many foods. In order to assess the botulism hazard, the effect of heat treatments at 70, 75, 80, 85, and 90 degrees C combined with refrigerated storage for up to 90 days on growth from 10(6) spores of nonproteolytic C. botulinum (types B, E, and F) in an anaerobic meat medium containing 2,400 U of lysozyme per ml (50 microg per ml) was studied. Provided that the storage temperature was no higher than 8 degrees C, the following heat treatments each prevented growth and toxin production during 90 days; 70 degrees C for >/=2,545 min, 75 degrees C for >/=463 min, 80 degrees C for >/=230 min, 85 degrees C for >/=84 min, and 90 degrees C for >/=33.5 min. A factorial experimental design allowed development of a predictive model that described the incubation time required before the first sample showed growth, as a function of heating temperature (70 to 90 degrees C), period of heat treatment (up to 2,545 min), and incubation temperature (5 to 25 degrees C). Predictions from the model provided a valid description of the data used to generate the model and agreed with observations made previously.

Botulinum Toxins↗
PubMed2000

Quantification of Clostridium botulinum toxin gene expression by competitive reverse transcription-PCR.

Clostridium botulinum produces a characteristic botulinum neurotoxin which can cause an often fatal neuroparalytic condition known as botulism. Although food-borne botulism is rare, critical screening by food companies is necessary to ensure that food products are safe. At present, the food industry assesses the risks of botulinum neurotoxin production by challenge testing to check any new food products and to check the efficacy of new storage regimes. Challenge testing involves artificial introduction of defined strains of microorganisms into food, and microbial growth and possible toxin production are then monitored. Botulinum toxin is normally analyzed by using the mouse bioassay. However, the mouse bioassay is expensive, slow, and politically sensitive because of animal rights issues. In this paper we describe adaptation of a new assay, competitive reverse transcription-PCR (RT-PCR), to monitor botulinum neurotoxin production. This method accurately measures the level of toxin-encoding mRNA in C. botulinum cells. Measurement of mRNA should provide a good indication of gene expression as mRNA is turned over rapidly in bacterial cells. In addition, the method is rapid, specific, and sensitive. The competitive RT-PCR method was developed to examine C. botulinum E VH toxin gene expression and was used to investigate the level of toxin production by C. botulinum E VH when the organism was grown in two different types of broth. The results which we obtained with the competitive RT-PCR method demonstrated that this method is more rapid and more sensitive than the mouse bioassay.

Animals↗
PubMed2004

Vaccination with recombinant whole heavy chain fragments of Clostridium botulinum Type C and D neurotoxins.

Mice and ducks were subcutaneously immunized with recombinant whole heavy (H) chains of Clostridium botulinum type C and D neurotoxins, which were expressed as glutathione S-transferase fusion proteins. In the case of mice, it was confirmed that two immunizations with type C- and D-H chains, 10 microg each time, significantly increased the specific antibodies against 100-kDa H chains of type C and D neurotoxins in an immunoblot analysis and an enzyme-linked immunosorbent assay, respectively. The mice immunized with type C- and D-H chains showed no symptoms of botulism when they were challenged with C- and D-16 S toxins at doses, given intraperitoneally, of up to 10(5) and 10(6) minmum lethal doses (MLD), respectively, per mouse. Ducks were immunized with a total of 100 microg of type C-H chain. The ducks also developed specific antibodies to the type C-H chain and showed significant protection against a challenge with 10(3) duck MLD of C-16 S toxin given intravenously. These results indicate that recombinant whole H chains can be used as an effective and safe vaccine for type C and D botulism in domestic animals.

Amino Acid Sequence↗
PubMed1991

Arm abscesses caused by Clostridium botulinum.

Wound botulism is an uncommon disorder that continues to be rarely reported in the United States. A 34-year-old intravenous heroin user was admitted to the Loma Linda, Calif., Veterans Administration hospital with multiple abscesses on his forearms. His clinical course was compatible with botulism, and his culture and serum were positive for Clostridium botulinum toxin type A. Early culture and/or serum identification can lead to prompt diagnosis, treatment, and improvement in the morbidity and mortality rates of this disease.

Abscess↗
PubMed2004

Natural Clostridium botulinum type C toxicosis in a group of cats.

Clinical signs of botulism were observed in a group of eight cats, four of which died, after being fed pelican carrion. Clostridium botulinum type C was isolated from one cat. The microorganism and its toxin were found in the pelican. This is apparently the first report of natural botulism in cats.

Animal Feed↗
PubMed2000

Fatal Clostridium botulinum toxicosis in eleven Holstein cattle fed round bale barley haylage.

Twenty-two lactating Holstein cattle in Tennessee had clinical signs of intoxication with preformed Clostridium botulinum toxin. These signs included weakness, paralysis of the tongue and chest muscles, abdominal breathing, and, in 11 of the 22 cows, death. Differential diagnoses included hypocalcemia, hypomagnesemia, carbohydrate overload, and several toxicoses including mycotoxin, lead, nitrate, organophosphate, atropine or atropine-like alkaloid, and botulism. A diagnosis of botulism by the ingestion of preformed C. botulinum type B toxin was made by eliminating these other diseases, by finding C. botulinum type B spores in 3 bales of round bale barley haylage fed to these cattle, and by isolating preformed type B toxin from 1 of the 3 bales. Confirmation of the toxin type was made by demonstrating mouse lethality by intraperitoneal injection of specimen extracts with neutralization by C. botulinum type B antitoxin. The haylage, harvested green and encased in black plastic bags to facilitate fermentation, was presumably contaminated by the botulinum toxin when fermentation failed to produce enough acid to lower the pH to 4.5, the pH below which C. botulinum growth is inhibited. Farmers and ranchers who use round hay balers to produce haylage should be alert to this potential problem.

Animal Feed↗
PubMed2026

Isolation and Characterization of Botulinum Neurotoxin-Producing Clostridium caccae from Meat and Soil - China, 1990-2025.

WHAT IS KNOWN ABOUT THIS TOPIC?: Botulinum neurotoxin (BoNT)-producing clostridia are categorized into six physiologically and phylogenetically distinct groups (Groups I-VI). Increasing genomic evidence indicates that bont genes are distributed across multiple clostridial lineages, highlighting the taxonomic complexity of BoNT-producing clostridia. Clostridium caccae (C. caccae), first described in 2020 from human stool, was originally described as a strain lacking the bont gene; however, its association with BoNT production has not been previously recognized. WHAT IS ADDED BY THIS REPORT?: Six isolates originally identified as Group II C. botulinum in China were assigned to C. caccae based on the average nucleotide identity (ANI) analysis. In addition, 140 publicly available genomes of C. caccae were analyzed. Among the 146 C. caccae strains, 63% carried bont genes (subtypes B4, E9, E12, and F6), with geographical distribution patterns differing by subtype. WHAT ARE THE IMPLICATIONS FOR PUBLIC HEALTH PRACTICE?: A subset of Group II C. botulinum strains should be considered as C. caccae, indicating that this species represents a previously under-recognized reservoir of multiple BoNT subtypes. To our knowledge, this is the first study to link BoNT-producing C. caccae to both soil contamination and human botulism, highlighting that BoNT-producing C. caccae may be an under-recognized contributor to the risk of botulism.

Botulinum neurotoxin↗
PubMedDate not supplied

[Presence of botulinum-producing clostridia in the soils of Entre Ríos].

Botulism, well known for centuries, continues to preoccupy the Public Health authorities, food and agricultural industries. In its oldest form it results from ingestion of food containing botulinum toxin. To correctly evaluate the incidence of botulism it is fundamental to know the distribution of botulinum toxin-producing clostridia (Clostridium botulinum, C. baratii, C. butyricum and C. argentinense) in nature, specially in the soil, its main reservoir. In our country, this study has been carried out in a partial way. With the aim to contribute to such knowledge, 240 soil samples from Entre Ríos province were examined for C. botulinum and 35 (14.6%) samples resulted positive.

Animal Husbandry↗
PubMed1978

Foodborne hazards of microbial origin.

Foods can serve as vehicles of many pathogenic and toxigenic agents of disease. Bacterial agents comprise three groups: 1) those that grow in the food and produce an active toxin before consumption (e.g., clostridium botulinium); 2) those that merely exist as contaminants in the food but are able to initiate infection when swallowed (e.g., Salmonella spp.); and 3) those that multiply and produce large numbers of vegetative cells in the food, then release an active enterotoxin when they sporulate in the gut. A few parasitic (e.g., Trichinella spiralis) and viral agents (e.g., hepatitis A) also can be transmitted by food. Botulinum poisoning is the deadliest foodborne disease. The potential danger of botulism from cured meats is a major factor in the argument over use of nitrites as meat curing agents. A new disease called infant botulism has been recognized since 1976. Apparently it is not foodborne but results from intraintestinal growth of C. botulinum in very young infants. Salmonellosis is the most important of the foodborne diseases from the standpoint of overall human health. The primary vehicles are contaminated raw meat, poultry, and eggs. Faulty food handling practices are responsible for most food poisoning in the United States.

Bacillus cereus↗
PubMed1990

[3 cases of Clostridium botulinum type C intoxication in the dog].

Three cases of dogs showing symptoms of acute lower motor neurone disease are reported, in which a diagnosis of type C botulism was established. In one of the dogs, botulism was believed to be due to type C toxin-containing carcases in poultry droppings; in the other two, causative relationship with poultry was extremely suggestive. The toxin was shown to be present in the serum on days one and three. Clostridium botulinum was still present in cultures of the faeces within sixteen days after onset of the symptoms.

Animals↗
PubMed1985

Differential effect of botulinal toxin on esophageal motor function in infants.

The effect of Clostridium botulinum toxin on esophageal motor function was studied in four infants (ages 5-9 months) with confirmed infant botulism. Esophageal motility studies using a perfused catheter assembly were performed during the acute phase in all patients, and during the recovery phase in one patient. Motor function of the proximal esophagus and upper esophageal sphincter was abnormal in each, while motor function of the distal esophagus and lower esophageal sphincter was normal. Mean lower esophageal sphincter pressure was 24 mm Hg for the group (normal: 15-30 mm Hg). Sequential studies of proximal esophageal motility in one infant revealed a return of normal motor function which correlated with recovery of peripheral muscle strength and gag reflexes. C. botulinum toxin impairs proximal, but not distal, esophageal motor function in the infant botulism model. This effect appears to be consistent with the known action of the toxin on synaptic acetylcholine release and current concepts regarding distribution of cholinergic and noncholinergic neurotransmitter receptors in the esophagus.

Botulinum Toxins↗