Cause Of Botulism Research Articles

Exploring the genetic background of the botulism neurotoxin BoNT/B2 in Spain.

To determine whether the neurotoxin BoNT/B2 causing botulism in Spain is clonal, the genetic diversity and phylogenetic relationships of Clostridium botulinum from food-borne episodes and infant cases of the condition were explored. The botulinum toxin gene (bont) subtype, the variable region of the flagellin gene (flaVR), and a seven-gene multi-locus sequence type were examined by sequencing 37 BoNT-positive cultures obtained over the period 2010 to 2022. Out of 37 botulism events, 16 food-borne episodes and 16 infant cases were associated with bont/b2. Eight bont/b2 alleles were detected [nucleotide distance range 0.0259-0.415%, Hunter and Gaston discrimination index (HGDI) 0.71]. The most common bont/b2 allele corresponded to that of strain Prevot 25 NCASE and its single and double locus variations (87.5%). Four known flaVR types were identified (HGDI 0.79), along with one previously unknown (flaVR-15). Sixteen sequence types (STs) (HGDI 0.89) were recorded including seven new STs (ST164-ST170; 10 new alleles) and five new STs (ST171-ST175; with new allele combinations) were also noted. Correlations among some STs and flaVR types were seen. Overall, the present results show that the combined analysis of bont/b2-flaVR-ST at the nucleotide level could be used to track botulism events in Spain. The neurotoxin BoNT/B2 has largely been responsible for human botulism in Spain. The polymorphism analysis of bont/b2, flaVR typing, and sequence type determinations, revealed a wide variety of clones to be responsible for human botulism, ruling out a common source of acquisition. IMPORTANCE Botulism, a potentially fatal disease, is classically characterized by a symmetrical descending flaccid paralysis, which if left untreated can lead to respiratory failure and death. Botulinum neurotoxin (BoNT), produced by certain species of Clostridium, is the most potent biological toxin known, and the direct cause of botulism. This study characterizes the acquisition in Spain of two forms of botulism, i.e., food-borne and infant botulism, which are largely caused by the main neurotoxin BoNT/B2. Polymorphism analysis of the bont/b2 gene, typing of the flagellin variable region sequence (flaVR), and multilocus sequence typing, were used to explore the genetic background of Clostridium botulinum group I. To our knowledge, this is the first phylogenetic and typing study of botulism undertaken in Spain.

Potential Risk of Botulinum Neurotoxin -producing Clostridia Occurrence in Canned Fish.

Heat treatment is indispensable in fish canning to provide an acceptable shelf life. Its optimisation reduces the risk of the presence of Clostridium botulinum spores, which could potentially cause botulism cases. This study evaluated canned fish samples for botulism neurotoxin (BoNT)-producing clostridia contamination and can bulging through microbiological contaminant growth. A new analytical approach was developed for detection of such clostridia and phenotypically similar species. A total of 70 canned fish samples suspected of exhibiting bulging features were analysed. Culture methods were used to detect clostridia. The isolates obtained were evaluated on the basis of the exhibited phenotypic characteristics. Also, PCRs were used for the detection of genes determining BoNT production (non-toxic non-haemagglutinin (ntnh) genes) and the amplification of conservative 16S rDNA genes, which were Sanger sequenced. The obtained sequences were analysed using the Basic Local Alignment Search Tool. Clostridium genus species were isolated from 17 (24%) bulging and organoleptically changed samples. No ntnh genes were present in these isolates; however, sequencing confirmed the presence of C. sporogenes, a species with close affinity to C. botulinum. To eliminate the threat of foodborne botulism, laboratory diagnostic techniques must detect species of the Clostridium genus and elucidate their ability to produce BoNTs. Although Clostridium botulinum is the most common cause of botulism, the possibility may not be ignored that non-pathogenic Clostridium species may acquire botulinum toxigenicity. The similarity between the isolated strains of C. sporogenes and C. botulinum should be incorporated in the optimisation of heat treatment to guarantee a sterilised, microbiologically safe product.

Botulinum toxin: From the natural cause of botulism to an emerging therapeutic in veterinary medicine

Botulinum toxins are one of the most potent biological toxins known. They cause botulism in wildlife and livestock, and thus, are regarded as a problem in veterinary medicine. On the other hand, use of purified low dose pharmaceutical grade botulinum toxin serotype A (BoNT-A) in different human clinical implications such as movement, autonomic disorders, pain conditions have expanded in the last 30-40 years based on its long-lasting effects and a good safety profile. However, despite that, its use in veterinary practice is lagging far behind the already well-established use in humans. BoNT-A is not licensed for use in any veterinary medical condition, and overall, there are only few clinical trials and several case studies, the most notable being case reports and limited clinical studies of osteoarthritis and perioperative pain in dogs, lameness in horses, as well as the spasticity treatment in cats. Indications for BoNT-A use in veterinary medicine happen to be very similar when compared to human indications, which is not surprising due to pathophysiological similarities. This warrants further clinical research of novel indications of BoNT-A in veterinary medicine.    

Paper-based electrochemical peptide sensor for on-site detection of botulinum neurotoxin serotype A and C

Botulinum neurotoxins (BoNTs) produced by soil bacterium Clostridium botulinum are cause of botulism and listed as biohazard agents, thus rapid screening assays are needed for taking the correct countermeasures in a timely fashion. The gold standard method relies on the mouse lethality assay with a lengthy analysis time, i.e., 2–5 days, hindering the prompt management of food safety and medical diagnosis. Herein, we propose the first paper-based antibody-free sensor for reliable and rapid detection of BoNT/A and BoNT/C, exploiting their cleavage capability toward a synthetic peptide able to mimic the natural substrate SNAP-25. The peptide is labelled with the electroactive molecule methylene blue and immobilized on the paper-based electrode modified with gold nanoparticles. Because BoNT/A and BoNT/C can cleave the peptide with the removal of methylene blue from electrode surface, the presence of these neurotoxins in the sample leads to a signal decrease proportional to BoNT amount. The biosensor developed with the selected peptide and combined with smartphone assisted potentiostat is able to detect both BoNT/A and BoNT/C with a linearity up to 1 nM and a detection limit equal to 10 pM. The applicability of this biosensor was evaluated with spiked samples of orange juice, obtaining recovery values equal to 104 ± 6% and 98 ± 9% for 1 nM and 0.5 nM of BoNT/A, respectively.

Foodborne Botulism in Ukraine from 1955 to 2018.

We aim to identify possible biological, social, and economic factors that could influence the prevalence of foodborne botulism (FB). The objective of this article is to assess epidemiological peculiarities of FB in Ukraine from 1955 to 2018 using national epidemiological surveillance data. This article presents an epidemiological descriptive population-based study of the epidemiology of FB using correlation analysis. From 1955 to 2018, 8614 cases of botulism were recorded in Ukraine causing 659 deaths. The distribution of types of botulism toxins is represented by type A (7.97%), B (59.64%), suspected as C (0.56%), E (25.47%), others (5.33%), and unidentified (1.04%). From 1990 to 2015, the rate correlation between Human Development Index (HDI) and incidence of botulism was −0.75 ± 0.20. Homemade canned meat and fish continue to be the leading causes of botulism in Ukraine. Cases related to commercial food were rare or absent, but in recent years (2017–2018), their percentage has increased to 32.56%. The HDI and botulism have an inverse mathematical correlation and predictable logical relationship: with an HDI increase, the incidence of FB decreased. In general, food botulism in Ukraine is related to traditional socioeconomic factors related to cultural food habits. In the face of declining living standards and uncertainty that food products will be physically or economically available, homemade preservation increases. Home food preservation is a major cause of botulism in Ukraine. The elimination of FB is possible in Ukraine only with the complete cessation of home canning and state control over the manufacture and sale of commercial canned products.

An Integrated Pharmacophore/Docking/3D-QSAR Approach to Screening a Large Library of Products in Search of Future Botulinum Neurotoxin A Inhibitors.

Botulinum toxins are neurotoxins produced by Clostridium botulinum. This toxin can be lethal for humans as a cause of botulism; however, in small doses, the same toxin is used to treat different conditions. Even if the therapeutic doses are effective and safe, the adverse reactions could be local and could unmask a subclinical impairment of neuromuscular transmissions. There are not many cases of adverse events in the literature; however, it is possible that sometimes they do not occur as they are transient and, if they do occur, there is no possibility of a cure other than to wait for the pharmacological effect to end. Inhibition of botulinum neurotoxin type A (BoNT/A) effects is a strategy for treating botulism as it can provide an effective post-exposure remedy. In this paper, 13,592,287 compounds were screened through a pharmacophore filter, a 3D-QSAR model, and a virtual screening; then, the compounds with the best affinity were selected. Molecular dynamics simulation studies on the first four compounds predicted to be the most active were conducted to verify that the poses foreseen by the docking were stable. This approach allowed us to identify compounds with a calculated inhibitory activity in the range of 316–500 nM.

Early recovery of botulism: one decade of experience

Background Botulism is a rare but serious disease, which appears in different forms. In this study, we reviewed the clinical features, laboratory data, and outcomes of patients who referred to our tertiary center. Materials and methods All confirmed cases of botulism referred to an academic referral center and a teaching hospital during 2009–2019, were retrospectively reviewed. Results Fifty-three cases of clinical or laboratory-confirmed botulism were examined in this study. Nineteen patients were confirmed by laboratory data (serotype A (89.5%) and serotype E (10.5%)). In seven cases, the cause of botulism was unclear. In two patients, systemic symptoms emerged after the therapeutic injection of botulinum neurotoxin. The majority of cases (83%) were caused by an obvious food source. In 66% of cases, the initial symptoms emerged within less than 36 h, while in 20.8% of cases, the symptoms developed within or after 36 h; however, in seven patients that their botulism sources were unclear, the onset could not be estimated. All patients showed cranial involvement and generalized manifestation, and 49.1% had gastrointestinal symptoms. Except for two patients who were not treated due to immediate drug reactions who manifested severe hemodynamic instability, the rest of the patients were treated with trivalent antitoxin (A, B, and E). The complete resolution of the symptoms during hospitalization was documented in 50.9% of the patients. About 17% of the patients were intubated. Two patients died due to massive bilateral pulmonary thromboembolism and cardiac asystole following respiratory failure. Conclusions Although the complete resolution of the symptoms usually takes several weeks, in our experience, most patients showed at least partial resolution upon discharge. Early treatment results in better outcomes.

Suboxone abuse with shared needle as a cause of Botulism in an IV drug user (P5.9-027)

Suboxone abuse with shared needle as a cause of Botulism in an IV drug user (P5.9-027)

Computational method yields therapeutic ‘mini proteins’

Scientists have used a computational method to design brand-new small proteins that inhibit specific therapeutic targets such as the hemagglutinin protein involved in flu infection. The findings suggest that this method—de novo protein design—could be a wave of the future for drug discovery. Researchers often find drugs by screening large collections of compounds for those that happen to hit biological targets. Using computers to intentionally design compounds to hit targets is a newer and less proven strategy. A key advantage of this route to drug discovery is that its underlying prediction algorithms can be improved over time, whereas screening is essentially random. David Baker of the University of Washington and coworkers demonstrated the potential of this new approach by designing therapeutic “mini proteins,” each containing about 40 amino acids (Nature 2017, DOI: 10.1038/nature23912). In cell culture experiments, the proteins inhibited hemagglutinin and botulinum neurotoxin B, the cause of botulism. And

Black tar heroin skin popping as a cause of botulism (P3.178)

Black tar heroin skin popping as a cause of botulism (P3.178)

Three outbreaks of foodborne botulism caused by unsafe home canning of vegetables--Ohio and Washington, 2008 and 2009.

Foodborne botulism is a potentially fatal paralytic illness caused by ingestion of neurotoxin produced by the spore-forming bacterium Clostridium botulinum. Historically, home-canned vegetables have been the most common cause of botulism outbreaks in the United States. During 2008 and 2009, the Centers for Disease Control and Prevention (CDC) and state and local health departments in Ohio and Washington State investigated three outbreaks caused by unsafe home canning of vegetables. We analyzed CDC surveillance data for background on food vehicles that caused botulism outbreaks from 1999 to 2008. For the three outbreaks described, patients and their family members were interviewed and foods were collected. Laboratory testing of clinical and food samples was done at the respective state public health laboratories. From 1999 to 2008, 116 outbreaks of foodborne botulism were reported. Of the 48 outbreaks caused by home-prepared foods from the contiguous United States, 38% (18) were from home-canned vegetables. Three outbreaks of Type A botulism occurred in Ohio and Washington in September 2008, January 2009, and June 2009. Home-canned vegetables (green beans, green bean and carrot blend, and asparagus) served at family meals were confirmed as the source of each outbreak. In each instance, home canners did not follow canning instructions, did not use pressure cookers, ignored signs of food spoilage, and were unaware of the risk of botulism from consuming improperly preserved vegetables. Home-canned vegetables remain a leading cause of foodborne botulism. These outbreaks illustrate critical areas of concern in current home canning and food preparation knowledge and practices. Similar gaps were identified in a 2005 national survey of U.S. adults. Botulism prevention efforts should include targeted educational outreach to home canners.

Generalized botulism: diagnostic problems in a patient treated with botulinum toxin type A

Editor, Botulinum toxin type A (BT/A) is commonly used nowadays in the treatment of patients with localized muscle spasticity. The toxin is considered to be effective and safe in therapeutic doses, but cases of generalized botulism have been described, including the occurrence of a botulism-like syndrome after intramuscular injections of BT/A (Bakheit et al. 1997; Pang & O'Day 2006). Botulinum toxin is the most effective method of treatment for patients with localized muscle spasticity and can restore function sufficiently to allow patients to resume normal life. We present the case of a 67-year-old woman treated with BT/A for blepharospasm. On 26 July 2002 the patient received an injection of 125 units (1LD50) of BT/A (Dysport Clostridium; IPSEN Biopharm Ltd, ) from a 500-unit vial, administered to the orbicularis oculi muscle in the left eye. On 4 August the subject experienced nausea, vomiting, general weakness, xerostomia and hoarseness. The following day she experienced disturbances in her vision, photophobia, bilateral weakness of visual acuity and diplopia, followed later by abdominal meteorism and obstinate constipation. She also experienced problems in swallowing and disturbances in respiration. On 7 August 2002 the patient was admitted to the Department of Neurology, Medical University of Lublin. During physical examination the patient was conscious, sleepy and apathetic. Her body temperature was 36.7 °, blood pressure 95/65 mmHg and pulse 84 beats/min. During examination, the patient appeared pale and expressionless. Mydriasis, asymmetric pupils, no pupillary reaction to light, accommodation disturbances and impaired ocular movement with bilateral ptosis, especially on the left, were noted. Lack of palatine reflexes was also observed, along with xerostomia and shallow respiration with limited movement of the chest and diaphragm. The patient's breathing frequency was 24 breaths/min. Her heart rate was rhythmic, but the cardiac sound was dull. Moreover, a distended belly, suppression of peristaltic movement, full urinary bladder and lack of diuresis were observed. Muscle power in the limbs was preserved, along with hypotonicity and weakened reflexes. On the basis of this clinical picture, botulism was diagnosed. Blood tests to identify the presence of botulinum toxin were performed to confirm the diagnosis. Clinical improvement was achieved after administration of 150 ml botulinum antitoxin (polyvalent equine serum ABE composed of antitoxin botulinum A [5000 I.U.], antitoxin botulinum B [5000 I.U.] and antitoxin botulinum E [1000 I.U.] in a 10-ml vial). In laboratory diagnostics, during biological tests on mice with antitoxin neutralization, botulinum toxin type B was found to be present, which excluded BT/A Dysport as the cause of intoxication. On 16 August 2002, the 71-year-old sister of the patient was admitted to the Department of Infectious Diseases. She showed typical clinical symptoms of botulism, including symmetrical descending flaccid paralysis with prominent diplopia, dysarthria, dysphonia and dysphagia. Her initial symptoms, including xerostomia and signs of dyspepsia, had appeared 7 days after she had consumed home-preserved meat. Subsequently, it transpired that both sisters had eaten the preserved meat. In the second sister's case, biological tests on mice also confirmed the presence of botulinum toxin type B. Initial symptoms of botulinum intoxication usually appear within a time period ranging from 6 hours to 8 days; incubation time has been shown to correlate with the severity of the clinical course (Hughes et al. 1981; Cherington 1998; Shapiro et al. 1998). In the first case described here, the incubation period was short (10 hours), but in the patient's sister it was 7 days and thus it was impossible to exclude intoxication by the type A toxin after injection of BT/A. Cases of generalized botulism after BT/A have been described in the literature but after higher doses of 250–500 units (Bakheit et al. 1997). Nevertheless, the cardinal cause of botulism is the consumption of food infected with botulinum toxin. In the USA 50% of food botulism cases are caused by toxin type A, 25% by toxin type B, and the remaining cases by toxin type E (Shapiro et al. 1998). In Poland, where the most frequent cause of botulism is toxin type B, an average of 82 cases of illness have been noted per year since 1998, which is the highest in the EU (Therre 1999).

Microbiological aspects and technological need: technological needs for nitrates and nitrites.

Nitrate and nitrite are used in meat and fish curing, and in the manufacture of certain cheeses. Nitrate itself has little antimicrobial effect and in most applications could be replaced by lower concentrations of nitrite. Further, improved hygiene diminishes the need for nitrite. The antimicrobial activity and technological needs for nitrate and nitrite are reviewed. It is concluded that the technological needs for nitrite in meat products stored at < 10 degrees C could be met by added nitrite concentrations of 50 mg/kg. The overall effect of nitrate in salted fish appears to be marginal. In such products, Vibrio parahaemolyticus does not grow at salt concentrations of 10% and food poisoning by this organism is not related to the absence of nitrate or nitrite; growth of Clostridium botulinum Type E (the predominant cause of botulism from fish products) is arrested by salt concentrations of 3-4%. Listeria monocytogenes in seafood cannot be controlled by nitrite. The use of nitrate in cheese production could be avoided, or at least reduced to a low level by avoiding silage with a high count of gas-producing Clostridia and hygienic milk collection.

Botulism in Commercially Canned Foods

Occasionally, a problem thought to be well under control returns to plague us. This is the case with botulism in commercially canned foods. These foods have had a remarkably good record over the last 45 years with approximately 775 billion cans of commercially canned foods being consumed with only four known deaths through 1971. Beginning in 1971, however, botulinal toxin and/or Clostridium botulinum has been found in commercially canned vichyssoise, chicken vegetable soup, peppers, marinated mushrooms, tuna fish, beef stew, and in 41 cans of mushrooms from 20 lots packed by seven domestic and two foreign producers. The typical cause of botulism in canned foods is underprocessing which may result from inadequate equipment, improper operating procedures, and scheduled processes which are not appropriate for the actual operating conditions being used.

Wound Botulism

A wound infection contaminated with Clostridium botulinum is a little-appreciated cause of botulism, and may be more common than the literature suggests.