Clostridium Botulinum Research Articles

From genome inspection to clinical significance: source attribution of a foodborne botulism outbreak

Abstract Background Foodborne botulism is a neuro-paralytic disease caused by the ingestion of botulinum neurotoxins (BoNTs). Given its high fatality rates and economic consequences, prompt management of human cases requires collaboration among public health authorities, supporting hospitals in rapid diagnosis, food inspections, and formal analysis. This study aims to investigate the food source responsible for a botulism outbreak in Northern Italy, which resulted in the hospitalization of a person. Methods Faecal specimen from the patient, and suspected food were collected (industrial-made asparagus cream, both packages consumed and still closed). BoNT presence and identification was performed by mouse-test. After the enrichment in TPGY broth, samples were plated on CMM medium to observe the proteolysis, and on EYA medium for colonies isolation. DNA was extracted and genome was sequenced by Illumina MiSeq. Typing was performed by MLST. SNP-based phylogenetic analysis was conducted, including isolated strains from outbreaks occurred in the same year and geographic area. Results The strains were identified as group I proteolytic Clostridium botulinum, with BoNTB responsible for the outbreak. Faeces and the ingested food exhibited the same Sequence Type (ST178), differing by only one SNP. Notably, the closed package had a distinct ST (ST29), and a significant divergence from the outbreak strains (7282 SNPs). Conversely, the other strains displayed SNP differences ranging from 11116 to 46536. Conclusions Genome characterization enabled the identification of the outbreak source as the ingestion of BoNTB present in asparagus cream, suggesting also the circulation of several different strains of Cl. botulinum causing foodborne outbreaks. These findings highlight the ongoing public health concern of foodborne botulism, even in industrial-processed foods. Our study underscores the importance of source attribution and effective outbreak management in preventing hazardous botulism outbreaks. Key messages • Genome sequencing enabled the source identification of a foodborne outbreak caused by Cl. botulinum toxin B. • Our study emphasizes the current significance of foodborne botulism as a public health issue.

BOTULIN TOXIN AND ITS USE IN THE TREATMENT OF BRUXISM: a narrative review of the literature

The purpose of this paper is to review the literature in narrative form on the use of Botulinum Toxin (BTX) in the treatment of bruxism. TXB is a substance produced from a bacterium known as Clostridium botulinum. Thus, the most widely used on the market today is type A, which in its applicability manages to promote muscle relaxation. Therefore, when correlating Bruxism, involuntary contraction of the masticatory muscles, it is clear that this dysfunction, which causes headaches, tooth wear, abfraction, among various other damages to the patient, can be associated with the use of TXB. In this context, the aim was to study up-to-date articles on the use of botulinum toxin in the treatment of bruxism. Therefore, its use in dentistry is highly effective, as it is a minimally invasive treatment applied by the dentist and has effective therapeutic results.

Bacteria Associated with Kitchen Surface in Some Selected Areas in Sokoto State, Nigeria

Contamination of kitchen utensils and their role as a source or microbial reservoir is now becoming a major concern in addressing food hygiene and safety. This study was carried out to isolate and identify bacteria associated with kitchen surfaces in some selected areas in Sokoto State, Nigeria. The swab sticks were collected from the Kitchen surface in Arkilla, Lowcoast, Mabera, Minnanata and Runjin Sambo. Samples collected were isolated and characterized using microscopy and biochemical tests. The most frequently isolated bacteria were Citrobacter freundi with (16%), followed by Staphylococcus cohnii with (12%). Clostridium fallax, Bacillus cereus, Salmonella sp., Staphylococcus saccharolyticus, Clostridium botulinum and vibrio cholera, had (8%) each. Clostridium septicum, Staphylococcus haemolyticus, Lactobacillus acidophilus, Staphylococcus intermedius, Salmonella para typhi A, Shigella specie, Staphylococcus xerosis, Clostridium butyricum had (4%) each. Also, the highest mean count of 1.18x105cfu/ml/50cm2 was observed from bacteria isolated in Arkilla area. The implication of this finding is that potentially pathogenic bacteria like Staphylococcus, Salmonella, Shigella and Vibrio species may be of public health concern. The need for disinfection of kitchen and covering the utensils use in kitchen is now very important.

Food-Borne Diseases and their Impact on Health

Food safety remains a vulnerable issue in the face of foodborne disease outbreaks, which have significant consequences for individuals, the food industry, and the economy. This article aimed to investigate the impact of food borne diseases on food quality along with the public health concerns related to contaminated foods. This study was conducted through secondary data extracted from the literature on an emerging and concerning topic, which revealed that the emergence of antibiotic resistance in foodborne bacteria has a significant impact on public health. Resistant bacterial infections pose considerable costs to society and threats to food safety and the health of the population as a whole. Multidrug-resistant microorganisms causing illnesses possess several biological mechanisms to counteract the drugs’ effectiveness and avoid being killed by them. It has been extracted that foodborne illnesses are caused by bacterial infections from various pathogens, including Clostridium perfringens, Clostridium botulinum, Listeria monocytogenes, Vibrio spp., Shigella spp., Escherichia coli, Campylobacter spp., and Salmonella spp. Foodborne illness epidemiological surveillance methods are powerful tools for monitoring the phenomenon that occurs in populations. These population-based approaches, focused on advancing technology and advanced molecular subtyping available to public health laboratories, are essentially suitable for cooperation with foodborne diseases correlated with cluster-produced and widely distributed food products.

Application of metagenomics for diagnosis of broilers displaying neurological symptoms

BackgroundThirty-two-day-old broiler chickens at a farm located in northwestern South Korea displayed adverse neurological symptoms including limping, lying down, and head shaking. Approximately 2.1% of chickens died or were culled due to severe symptoms. Five carcasses were submitted to the Avian Disease Division of the Animal and Plant Quarantine Agency (APQA) for disease diagnosis.ResultsBroilers displayed severe pericarditis and perihepatitis associated with gross lesions. Broilers also displayed microscopic lesions in the cerebrum and in the granular layer of the cerebellum, which were associated with multifocal perivascular cuffing and purulent necrosis in the cerebrum, and severe meningitis with heterophil and lymphocyte infiltration. Staphylococcus spp. were identified in the liver and heart using bacteriological culture. PCR/RT-PCR assays revealed that broilers were negative for avian Clostridium botulinum, Newcastle disease virus, and avian encephalomyelitis virus. Bacterial and viral metagenomic analysis of brain sample further revealed the presence of Pseudomonas spp. and Marek’s disease virus, which are known etiological agents of chicken meningoencephalitis.ConclusionsThis study reports a diagnostic analysis of gross and histopathological lesions from 32-day-old broilers displaying unique neurological symptoms that revealed the presence of the several neurological diseases including meningoencephalitis. The causative agents associated with meningoencephalitis of broilers that had not been identified by routine diagnostic methods could be diagnosed by metagenomics, which proves the usefulness of metagenomics as a diagnostic tool for unknown neurological diseases in broilers.

Design and characterization of a multi-epitope vaccine against Clostridium botulinum A3 Loch Maree intoxication in humans

Clostridium botulinum Loch Maree expresses an extremely potent botulinum neurotoxin subtype, A3 causing botulism and several gastrointestinal disorders in mammals. Several recombinant vaccines have been developed for human botulism and no vaccine is currently available for the treatment of diseases caused by other virulence factors. Hence, we designed, constructed, and characterized a multi-epitope vaccine from new virulence proteins identified from this organism using an immunoinformatics approach. The vaccine construct used in this study was designed from 6B cell linear epitopes, 12 cytotoxic T cell lymphocyte epitopes, and 15 helper T cell lymphocyte epitopes, with a defensin adjuvant and adjusting linker sequences. A molecular modeling approach was used to model, refine, and validate the 3D structure of the vaccine construct. Molecular docking studies were performed to determine the stability of the molecular interactions between the vaccine construct and human toll-like receptor 7. The in silico molecular cloning was used to clone a codon-optimized synthetic vaccine gene in pCYB1 vector and expressed in Escherichia coli. The results of this study identified six new virulence proteins: peptidoglycan hydrolase, SCP-like extracellular protein, N-acetylmuramoyl-l-alanine amidase, putative membrane protein, drug/metabolite exporter, and bacillolysin. The top B-cell, cytotoxic T-cell lymphocyte, and helper T-lymphocyte epitopes were predicted from these virulence proteins with greater accuracy and reliability. HLA-A*02:01 and HLA-A*03:01 were identified as HLA-A-binding alleles for cytotoxic T-cell lymphocyte epitopes. DRB1*0110 and DRB1*0115 are the dominant alleles that bind to helper T-cell lymphocyte epitopes. The synthetic gene construct was highly expressed in a heterologous host and produced considerable amounts of antigenic protein. The multi-epitope vaccine is more conservative in the sequence-structure–function link, immunogenic with less allergenicity, and possibly provokes cellular and humoral immunity. The present study suggests that the designed multi-epitope vaccine is a promising prophylactic candidate for the virulence and intoxication caused by subtype A3 strains.

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.

Device-Physics Realization of ZnO-MWCNT Nanostructure-Based Field-Effect Biosensor for Ultrasensitive Simultaneous Genomic Detection of Foodborne Pathogens.

The complex and versatile interactions among the wide variety of the nanostructures and the target analytes have primarily limited the detailed investigation of the transduction mechanism of nanomaterial-assisted electrical signal-based biosensors despite their high sensitivity, low-cost, portability, and ease of deployment. Hence, no common ground is formed detailing the principle of operation, demanding a strong need for systematic examination instead of hit and trial. Therefore, a maiden mechanistic investigation has been carried out in this paper for a field-effect-based biosensor device relying on the energy band diagram and the surface potential profile. To demonstrate the experimental evidence and appreciate the importance of food safety, three hazardous foodborne pathogens (Proteus mirabilis, Escherichia coli, and Clostridium botulinum) have been detected herein. The biosensor device, built on a hydrothermally synthesized zinc oxide and MWCNT (ZnO-MWCNT) composite nanostructure, simultaneously incorporates three fairly specific ss-DNA probes. Furthermore, the unmet challenge of biosensor device variability is addressed through the optimum selection of operating voltage of the device via a unique "voltage-selection-algorithm". We believe that the rigorous experimentation and the insightful device-physics realization demonstrated in this work will pave the way for a future decisive biosensor platform.

IDENTIFICATION AND PHARMACOLOGICAL PROFILING OF PLANT PEPTIDES AS INHIBITORS OF CLOSTRIDIUM BOTULINUM NEUROTOXINS A AND B

Objective: Clostridium botulinum causes botulism which is a neuroparalytic disease caused by neurotoxins that are caused all over the world. This disease is perceived as a food born disease caused by a potent neurotoxin. Plant peptides are those referred to as proteins at a length of small than 100 amino acids. Plants have antimicrobial properties present in almost all plant species. These plant peptides are classified based on their primary structures. Methods: Molecular docking was implemented to evaluate the efficacy of nine phytocompounds from C. botulinum against the target proteins (3nf3, 2npo). PyRx, a Virtual Screening software was utilized for molecular docking which allowed the inspection of three-dimensional protein structures and the identification of potential binding sites. Result: On analyzing the molecular docking result of puroindoline, impatiens exhibited the best binding affinity toward the two target proteins. Conclusion: The safe and efficient treatment for some facial pleats and lines is botulinum toxin. High levels of patient and physician satisfaction are linked to its use. This paper has extensively revised three aspects the binding activity, models, and their phytochemical properties. The most significant results involve solvent interactions, van der Waals interactions, electrostatic interactions, and hydrogen bonding were analyzed and concluded in this research paper.

Clostridium botulinum C3bot mediated effects on cytokine-induced psoriasis-like phenotype in full-thickness skin model

Clostridium botulinum C3 exoenzyme (C3bot) exclusively inhibits RhoA, B and C by ADP-ribosylation and is therefore used as a cell-permeable tool for investigating the cellular role of these Rho-GTPases. Rho-GTPases represent a molecular switch integrating different receptor signalling to downstream cascades including transcriptional cascades that regulate various cellular processes, such as regulation of actin cytoskeleton and cell proliferation. C3bot-induced inhibition of RhoA leads to reorganization of the actin cytoskeleton, morphological changes, and inhibition of cell proliferation as well as modulation of inflammatory response. In this study, we characterized the C3bot-mediated effects on a full-thickness skin model exhibiting a psoriasis-like phenotype through the addition of cytokines. Indeed, after the addition of cytokines, a decrease in epidermal thickness, parakeratosis, and induction of IL-6 was detected. In the next step, it was studied whether C3bot caused a reduction in the cytokine-induced psoriasis-like phenotypes. Basal addition of C3bot after cytokine induction of the full-thickness skin models caused less epidermal thinning and reduced IL-6 abundance. Simultaneous basal incubation with cytokines and C3bot, IL-6 abundance was inhibited, but epidermal thickness was only moderately affected. When C3bot was added apically to the skin model, IL-6 abundance was reduced, but no further effects on the psoriasis-like phenotype of the epidermis were observed. In summary, C3bot inhibits the cytokine-induced expression of IL-6 and thus may have an impact on the pro-inflammatory immune response in the psoriasis-like phenotype.

Evaluation of Aluminium Hydroxide Nanoparticles as an Efficient Adjuvant to Potentiate the Immune Response against Clostridium botulinum Serotypes C and D Toxoid Vaccines.

Clostridium botulinum serotypes C and D cause botulism in livestock, a neuroparalytic disease that results in substantial economic losses. Vaccination with aluminium-based toxoid vaccines is widely used to control the spread of botulism. Aluminium-based adjuvants are preferred owing to their apparent stimulation of the immune responses to toxoid vaccines when compared to other adjuvants. The aim of our study was to evaluate aluminium hydroxide nanoparticles as a potential substitute for alhydrogel in the botulism bivalent vaccine. Botulism vaccines were formulated with either alhydrogel or nanoalum and comparative efficacy between the two formulations was conducted by evaluating the immune response in vaccinated guinea pigs. A significant increase in immunological parameters was observed, with the antibody titres higher in the serum of guinea pigs (20 IU/mL of anti-BoNT C/D) injected with nanoalum-containing vaccine than guinea pigs inoculated with the standard alhydrogel-containing vaccine (8.7 IU/mL and 10 IU/mL of anti-BoNT C and anti-BoNT D, respectively). Additionally, the nanoalum-containing vaccine demonstrated potency in a multivalent vaccine (20 IU/mL of anti-BoNT C/D), while the standard alhydrogel-containing vaccine showed a decline in anti-BoNT C (5 IU/mL) antibody titres.

The effect of form and method of flavoring on microbiota of olive oil.

Flavoring olive oil is an increasing trend in olive oil processing. Growing consumer interest in flavored olive oils by natural material brings the need to evaluate the key limiting factors which is its microbiological stability. The present research compares the microbiological quality of olive oil flavored by 3 flavors (rosemary, garlic, and lemon), prepared by 3 methods to determine changes during storage. The comprehensive microbiological analyses (total number of microorganisms [TCM], anaerobic sporulates, yeasts, molds, bacteria of the family Enterobacteriaceae, bacteria of the genus Salmonella spp., Clostridium botulinum, and lactic acid bacteria) were conducted during 12 months of storage. The best results in TCM were observed in the oil flavored by fresh garlic (0.24 log CFU/mL). The highest counts of anaerobic sporulates were detected in the dried rosemary olive oil (1.10 log CFU/mL). The flavoring materials have significantly higher counts of microorganism than flavored oils (p < .05). The obtained results demonstrated that microorganisms are capable to survive in flavored olive oil and the method of flavoring can affect their growth in a selective way according to the chemical characteristics.

Septic shock due to Clostridium botulinum: a case report

BackgroundClostridium botulinum is an anaerobic, Gram-positive, rod-shaped bacterium that forms spores and the neurotoxin botulinum. It is best known for its toxin-induced flaccid paralytic disease, which is deadly without correct treatment. In this report, we show a completely different clinical course with fatal outcome.Case presentationA 37-year-old African man born in Sierra Leone was admitted. After admission, his condition rapidly worsened due to severe septic shock and progressive multiorgan failure. No neurological signs were seen. Despite treatment with antibiotics, fluid resuscitation, and norepinephrine, the multiorgan failure deteriorated further and he died as a result. Blood and synovial fluid cultures showed Clostridium botulinum. No botulinum toxins were found.ConclusionThis is a rare case of fatal septic shock due to Clostridium botulinum-induced septic arthritis without any sign of the classic clinical syndrome of botulism.

Prions in Microbes: The Least in the Most.

Prions are infectious proteins that mostly replicate in self-propagating amyloid conformations (filamentous protein polymers) and consist of structurally altered normal soluble proteins. Prions can arise spontaneously in the cell without any clear reason and are generally considered fatal disease-causing agents that are only present in mammals. However, after the seminal discovery of two prions, [PSI+] and [URE3], in the eukaryotic model microorganism Saccharomyces cerevisiae, at least ten more prions have been discovered, and their biological and pathological effects on the host, molecular structure, and the relationship between prions and cellular components have been studied. In a filamentous fungus model, Podospora anserina, a vegetative incomparability-related [Het-s] prion that directly triggers cell death during anastomosis (hyphal fusion) was discovered. These prions in eukaryotic microbes have extended our understanding to overcome most fatal human prion/amyloid diseases. A prokaryotic microorganism (Clostridium botulinum) was reported to have a prion analog. The transcriptional regulators of C. botulinum-Rho can be converted into the self-replicating prion form ([RHO-X-C+]), which may affect global transcription. Here, we outline the major issues with prions in microbes and the lessons learned from the relatively uncovered microbial prion world.

Experimental inoculation of the Botox perineurally of facial nerve and followed by histological assessment for its effect on the zygomatic bone of rabbits

Background: Botox, derived from the toxin produced by Clostridium botulinum, is an attenuated toxin employed as a treatment for cervical dystonia, overactive bladder, strabismus, cerebral palsy, and non-surgical cosmetic procedures. This study aimed to explore the impact of injecting Botox around the facial nerve on the left side of rabbits, specifically focusing on the histological changes in the zygomatic bone. Methods: Twenty-five adult rabbits of a local breed, weighing 1500-1800 gm and of both sexes, underwent a 15-day acclimation period prior to the experiment. The rabbits were randomly assigned to four groups (1st, 2nd, 3rd, and 4th). A singular intramuscular injection of Botox at a dose of 3.5 U/kg was administered to the left facial muscles. The rabbits in each group were then sacrificed at distinct time points after injection: day 10, day 15, day 30, and day 45. All rabbits survived until the end of the experiment, at which point they were euthanized with chloroform. Post-euthanasia, the rabbits' facial zygomatic bones were dissected and subjected to histological assessment. Results: The results indicated notable changes in the zygomatic bone, including prominent vacuoles within the bony matrix, cracks with cellular debris, limited osteocytes, and collagen bundles resembling woven bone. Additionally, in the day 30 group (T3), irregular bone borders and atrophied osteogenic cells were observed, while the day 45 group (T4) exhibited necrotic areas and bone fragments within the bone matrix. Conclusion: In conclusion, Botox, derived from Clostridium botulinum, serves various medical purposes. The study focused on injecting Botox around rabbits' facial nerves to assess its impact on zygomatic bones histologically. Following a controlled injection and specific time intervals, the examined bone samples exhibited distinct changes, providing insights into Botox's effects on bone morphology.

Exploring the potential of cold plasma therapy in treating bacterial infections in veterinary medicine: opportunities and challenges.

Cold plasma therapy is a novel approach that has shown significant promise in treating bacterial infections in veterinary medicine. Cold plasma possesses the potential to eliminate various bacteria, including those that are resistant to antibiotics, which renders it a desirable substitute for traditional antibiotics. Furthermore, it can enhance the immune system and facilitate the process of wound healing. However, there are some challenges associated with the use of cold plasma in veterinary medicine, such as achieving consistent and uniform exposure to the affected area, determining optimal treatment conditions, and evaluating the long-term impact on animal health. This paper explores the potential of cold plasma therapy in veterinary medicine for managing bacterial diseases, including respiratory infections, skin infections, and wound infections such as Clostridium botulinum, Clostridium perfringens, Bacillus cereus, and Bacillus subtilis. It also shows the opportunities and challenges associated with its use. In conclusion, the paper highlights the promising potential of utilizing cold plasma in veterinary medicine. However, to gain a comprehensive understanding of its benefits and limitations, further research is required. Future studies should concentrate on refining treatment protocols and assessing the long-term effects of cold plasma therapy on bacterial infections and the overall health of animals.

Evaluation of Pachycrepoideus vindemiae and Muscidifurax raptor (Hymenoptera: Pteromalidae) as biological control agents of Piophila casei (Diptera: Piophilidae) in ham production facilities.

Ham products play a fundamental role in the Italian economy, and attention to the problems of this sector is essential. The products of this sector can be attacked by parasites, which can cause direct and indirect damage. Piophila casei (L.) (Diptera: Piophilidae) a cheese and meat parasite, is currently responsible for hygiene problems in ham factories. The trophic activity of this pest on the products causes serious direct damage and it is a vector of various bacteria, including Clostridium botulinum. Another risk is human ingestion of the larvae, which are resistant to gastric juices action, potentially causing intestinal myiasis. Insecticide use of any type is not allowed in aging rooms, so biological control can represent a potential alternative. In this study, we investigate quality parameters such as successful rate of parasitism (SP), degree of parasitism (DP), sex-ratio (SR), life-span (LS), and emergence rates (ER) of 2 pupal parasitoids of Diptera: Pachycrepoideus vindemiae (Rondani) (Hymenoptera: Pteromalidae), currently the only known pupal parasitoid of P. casei, and Muscidifurax raptor (Girault and Sanders) (Hymenoptera: Pteromalidae). Our research confirmed P. vindemiae efficacy to parasitize P. casei and reported, for the first time, M. raptor as a pupal parasitoid of this Piophilidae. ER for both parasitoids were low, thus affecting the DP and SP estimations. This could be explained by the feeding behavior of the parasitoid host. The strongly female-biased SR for P. vindemiae supported previous studies. LS results in our experiment are crucial for determining the timing of release.

Wide range applications of botulinum toxin in medicine - literature review

Introduction and purpose : Botulinum toxin is one of the most dangerous natural toxins. Botulinum toxin poisoning causes botulism. Botulism is a set of systemic symptoms resulting from flaccid muscle paralysis. The first symptomatology of toxin poisoning was described at the beginning of the 19th century. After more than 200 years, botulinum toxin is associated primarily with aesthetic medicine treatments, where it is used to temporarily improve the appearance, mainly by reducing mimic wrinkles. Aim of the study: The aim of the study is to collect information available in PubMed, Google Scholar, academic textbooks and characteristics of botulinum toxin medicinal products, especially regarding its wide use, mechanism of action, adverse events and contraindications. As a result, the presentation of the versatile use of botulinum toxin as a medicinal product in many fields of medicine. Brief description of the state of knowledge: Botulinum toxin is produced by the anaerobic bacteria Clostridium botulinum. Its mechanism of action consists in flaccid muscle paralysis. Most preparations with botulinum toxin are only registered for strictly defined aesthetic medicine treatments. Other registered indications for the use of medicinal products with botulinum toxin include overactive bladder, various types of spasticity and prevention of headaches. In addition, in medicine, botulinum toxin is commonly used off-label. Summary: Botulinum toxin is widely used not only in aesthetic medicine. Intramuscular injections of botulinum toxin are successfully used in various fields of medicine, such as: urology, neurology, ophthalmology, gastroenterology, dermatology, gynecology, otolaryngology, dentistry and even psychiatry.

The Use of Botulinum Toxin in Medicine: Safety and Efficacy Based on the Latest Research

Introduction and aim of the study: Botulinum toxin, produced by anaerobic bacteria of the Clostridium botulinum family, is one of the most potent organic compounds known to man. Despite its toxicity, it has found widespread use in medicine. From applications in aesthetic medicine, through the treatment of various disease entities, botulinum toxin has become an extremely valuable tool in the hands of doctors. The aim of this study is to review the latest research on the use of botulinum in medicine, with an emphasis on its safety and effectiveness. Methodology: The review work is mainly based on articles searched in the Pubmed and Google Scholar databases in the years 2016-2023. The study was conducted by reviewing keywords such as "botulin", "botulin toxin", "botulin treatment", "botulin aesthetic". Current state of knowledge: Botulinum is used in the treatment of such disease entities as hyperhidrosis, cervical dystonia, migraines, neurogenic bladder, as well as in the treatment of scars and wrinkles. The use of botulinum in medicine is safe, provided that it is used by qualified specialists and in appropriate doses. An area that requires further research is the impact of long-term botulinum therapy on the antibodies produced against the toxin. Additionally, an important area for further research is patient monitoring, appropriate dose adjustment and frequency of administration, which in turn can increase the effectiveness of treatment. Summary: Botulinum toxin is an effective tool in medicine, which has many applications. However, as with any drug, it requires appropriate use and understanding of potential side effects. Further research and education are key to fully exploiting its potential and assessing the risk associated with long-term use.

Crystal Structure of the Catalytic Domain of a Botulinum Neurotoxin Homologue from Enterococcus faecium: Potential Insights into Substrate Recognition.

Clostridium botulinum neurotoxins (BoNTs) are the most potent toxins known, causing the deadly disease botulism. They function through Zn2+-dependent endopeptidase cleavage of SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins, preventing vesicular fusion and subsequent neurotransmitter release from motor neurons. Several serotypes of BoNTs produced by Clostridium botulinum (BoNT/A-/G and/X) have been well-characterised over the years. However, a BoNT-like gene (homologue of BoNT) was recently identified in the non-clostridial species, Enterococcus faecium, which is the leading cause of hospital-acquired multi-drug resistant infections. Here, we report the crystal structure of the catalytic domain of a BoNT homologue from Enterococcus faecium (LC/En) at 2.0 Å resolution. Detailed structural analysis in comparison with the full-length BoNT/En AlphaFold2-predicted structure, LC/A (from BoNT/A), and LC/F (from BoNT/F) revealed putative subsites and exosites (including loops 1-5) involved in recognition of LC/En substrates. LC/En also appears to possess a conserved autoproteolytic cleavage site whose function is yet to be established.