Next Article in Journal
A Calibration Curve Implanted Enzyme-Linked Immunosorbent Assay for Simultaneously Quantitative Determination of Multiplex Mycotoxins in Cereal Samples, Soybean and Peanut
Next Article in Special Issue
Botulinum Toxin Type A to Improve Facial Symmetry in Facial Palsy: A Practical Guideline and Clinical Experience
Previous Article in Journal
Association between Uremic Toxin Concentrations and Bone Mineral Density after Kidney Transplantation
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Toxins
Volume 12
Issue 11
10.3390/toxins12110716
toxins-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Review

Toxemia in Human Naturally Acquired Botulism

by
Christine Rasetti-Escargueil
,
Emmanuel Lemichez
and
Michel R. Popoff
*
Unité des Toxines Bactériennes, UMR CNRS 2001, Institut Pasteur, 75015 Paris, France
*
Author to whom correspondence should be addressed.
Toxins 2020, 12(11), 716; https://doi.org/10.3390/toxins12110716
Submission received: 15 October 2020 / Revised: 7 November 2020 / Accepted: 10 November 2020 / Published: 13 November 2020
(This article belongs to the Special Issue Potency, Duration of Action, and Pharmacodynamics of Human Neurotoxins)
Versions Notes

Abstract

:
Human botulism is a severe disease characterized by flaccid paralysis and inhibition of certain gland secretions, notably salivary secretions, caused by inhibition of neurotransmitter release. Naturally acquired botulism occurs in three main forms: food-borne botulism by ingestion of preformed botulinum neurotoxin (BoNT) in food, botulism by intestinal colonization (infant botulism and intestinal toxemia botulism in infants above one year and adults), and wound botulism. A rapid laboratory confirmation of botulism is required for the appropriate management of patients. Detection of BoNT in the patient’s sera is the most direct way to address the diagnosis of botulism. Based on previous published reports, botulinum toxemia was identified in about 70% of food-borne and wound botulism cases, and only in about 28% of infant botulism cases, in which the diagnosis is mainly confirmed from stool sample investigation. The presence of BoNT in serum depends on the BoNT amount ingested with contaminated food or produced locally in the intestine or wound, and the timeframe between serum sampling and disease onset. BoNT levels in patient’s sera are most frequently low, requiring a highly sensitive method of detection. Mouse bioassay is still the most used method of botulism identification from serum samples. However, in vitro methods based on BoNT endopeptidase activity with detection by mass spectrometry or immunoassay have been developed and depending on BoNT type, are more sensitive than the mouse bioassay. These new assays show high specificity for individual BoNT types and allow more accurate differentiation between positive toxin sera from botulism and autoimmune neuropathy patients.
Keywords:
botulism; botulinum neurotoxin; Clostridium botulinum; toxemia; serum
Key Contribution: Botulism is a severe disease in humans that requires early diagnosis for the management of patients. Botulinum toxin detection in the patient’s sera is the most direct laboratory method to confirm clinical botulism and to provide toxin typing. Based on literature reports; toxemia is observed in about 70% of food-borne and wound botulism cases, and only in about 28% of infant botulism cases.

1. Introduction

Botulism is a neurological disease of man and animals which is characterized by flaccid paralysis and inhibition of secretions from glands controled by cholinergic innervation, such as salivary, lachrymal, and sweat glands. The flaccid paralysis leading to respiratory distress is responsible for death in the most severe cases. Botulism results from the blockade of neurotransmitter release, notably acetylcholine at the neuromuscular junctions and other cholinergic neurons, by potent neurotoxins called botulinum neurotoxins (BoNTs), which proteolytically cleave SNARE (soluble N-ethylmaleimide-sensitive-factor attachment protein receptor) proteins involved in the exocytosis of neurotransmitters [1,2,3]. BoNTs are synthesized by Clostridium species named Clostridium botulinum and more rarely by other Clostridium species such as toxigenic strains of Clostridium baratii, Clostridium butyricum, and Clostridium argentinensis. According to their immunological properties based on neutralization by specific antisera, BoNTs are currently divided into 9 toxinotypes (A, B, C, D, E, F, G, H, or H/A, or F/A, X) [1,4]. BoNT amino acid sequences deduced from bont gene sequencing revealed that each toxinotype is subdivided into an increasing number of subtypes defined as >2.6% amino acid variation. Thus, 41 BoNT subtypes and multiple variants have been reported [5,6,7]. More recently, genome comparison showed the presence of bont related sequences in a few non-clostridial strains such as bont/Wo or bont/I from Weisenella oryzae, a bacterium of fermented rice, bont/J (ebont/F or bont/En) from an Enterococcus faecalis strain in a cow, and Cp1 from Chryseobacterium piperi from sediment [1,8,9,10,11,12]. However, no active BoNT responsible for human or animal botulism has been reported in the non-clostridial strains. In addition, a BoNT-like neurotoxin (PMP1) specific of invertebrate (Anopheles mosquito) has been characterized from a Paraclostridium bifermentans strain [13]. Albeit all BoNTs induce the same toxicological effects resulting in flaccid paralysis, most BoNT types and subtypes share a similar mechanism of action but differ by interaction with distinct receptors and intracellular SNARE targets cutting at different cleavage sites [2,14,15].
According to BoNT acquisition, different forms of human botulism are described. Ingestion of preformed BoNT in food is responsible for food-borne botulism, which is the most common form of botulism in many countries. Foods which are non-acidified, no or minimally heated, and contaminated by C. botulinum spores, are at risk to allow the growth of C. botulinum and BoNT production during their storage even at low temperature [16,17]. Ingested C. botulinum spores might lead to a toxico-infection under certain circumstances by colonization of the intestinal tract and in situ BoNT production. Children under the age of 1 year are susceptible to intestinal colonization by C. botulinum and to develop infant botulism [18,19]. Perturbations or limited functionality of the developing and yet immature infant gut microbiota are considered as a main risk factor [20,21]. More rarely, botulism by intestinal colonization (also called intestinal toxemia botulism) occurs in children above 1 year and adults. Factors that might impair the regular intestinal microflora such as bowel surgery, bowel anomalies, antibiotics, chemotherapy, radiation, immunosuppressive drugs, and altered nutritional patterns have been reported to be predisposing factors for adult intestinal toxemia botulism [22]. Wound botulism results from a wound contaminated with C. botulinum spores and subsequent in situ growth of C. botulinum and BoNT production. Moreover, aerosolized BoNT induces rare cases of laboratory botulism [23]. Iatrogenic botulism has been reported with toxin overdoses for therapeutic or cosmetic use [24,25,26,27].
The paradigm of foodborne botulism is that the ingested BoNT, which is preformed in contaminated food, transits through the digestive tract, crosses the intestinal barrier, reaches the blood and lymphatic circulation and disseminates to the peripheral nervous system [28]. In in vitro culture or in contaminated food, BoNT is produced in a complex form (also called progenitor toxin) by association with non-toxic proteins, including notably the non-toxic non-hemagglutinin (NTNH) and hemagglutinin (HA) proteins [25,29,30,31]. A main role of the associated proteins is to prevent BoNT degradation from the stomach acidic pH and digestive proteases. Indeed, NTNH, which shares a similar structure with BoNT, forms an interlocked complex with BoNT which is highly resistant to low pH and protease inactivation [32,33]. The precise mechanism of entry through the intestinal barrier of BoNT and/or the whole BoNT complex is still unclear. Free BoNT is able to pass through intestinal epithelial cells by transcytosis. In addition, HAs have been found to bind to intestinal cells, to disrupt the E-cadherin intercellular junctions between enterocytes, and thus to facilitate the paracellular passage of BoNT complexes. However, according to the distinct C. botulinum toxinotypes, numerous BoNT complexes lack HAs but contain other non-toxic proteins (OrfX1, OrfX2, OrfX3, P47) that have not been observed to be involved in the uptake of BoNT from the digestive tract [28,34,35,36]. Therefore, subsequently to the absorption from the intestinal tract, the passage of BoNT in the blood circulation seems to be a hallmark of BoNT intoxication [2,28]. Thus, the demonstration of BoNT in biological samples such as serum allows the confirmation of clinical botulism [24].
Treatment of botulism is based on supportive care, including mechanical ventilation in severe cases and botulinum antiserum. Equine anti-botulinum sera are available for adult botulism, and human anti-BoNT/A and -BoNT/B immunoglobulins (BabyBIG®) have been developed for infant botulism [24,37,38,39,40]. Anti-BoNT antibodies are the only specific treatment of botulism. However, since anti-BoNT antibodies mainly neutralize the toxins prior to their binding to neuronal cells, serotherapy’s efficacy depends on the rapid administration of antibodies after the onset of symptoms, ideally <24 h [24]. Therefore, early diagnosis of botulism is critical for the management of botulism. Botulism cases are suspected on the basis of clinical symptoms and epidemiological aspects. Botulism confirmation is supported by laboratory tests that are required for botulism typing [41,42]. Initially, food-borne botulism was confirmed by BoNT detection in suspected food samples using experimental animals [40]. Thus, from 1940 to 1970 the diagnosis of botulism in France at Institut Pasteur was performed only by BoNT investigation in food [43]. However, it was observed that BoNT can be identified in the serum or blood samples from patients with clinical botulism. Before 1970, some publications reported the presence of BoNT in the sera of patients with botulism type B or E and more rarely type A. From 1905 to 1962, 12 reports demonstrated BoNT in the sera of 20 human cases of botulism up to 25 days after the ingestion of contaminated food (reviewed in [44]). Two additional observations in 1963 and 1965 showed toxin positive sera in 5 and 4 patients with botulism type E and B, respectively [44,45]. From 1970, the detection of BoNT in serum samples was more routinely used for the diagnosis of botulism. Since the BoNT levels that have been observed in human sera are generally low, a high sensitive method of BoNT detection is required. The mouse bioassay which is the standard procedure and allows the detection of a few pg of BoNT per mL, was used in almost all studies. BoNT typing is performed by neutralization with specific anti-BoNT type antibodies [42]. This review focusses on BoNT detection in sera from patients with naturally acquired botulism. Since BoNT detection in serum is the most direct way to confirm a diagnostic of botulism, it is important to know the parameters of botulinum toxaemia such as prevalence of toxaemia according to botulism forms, duration, serum toxin levels, in order to adapt the serum sampling according to the onset of the disease and to interpret possible false negative/positive results.

2. Detection of BoNT in the Sera of Patients with Food-Borne Botulism

Investigations of BoNT detection in the sera of patients with food-borne botulism have been reported mainly since the 1970s (Table 1). Overall, from 1456 reported testings of BoNT by the mouse bioassay in the sera of patients suspected of botulism, 633 (43%) were positive (Table 1). However, a high heterogeneity is observed between the different investigations. Some of them concerned a small number of case reports or small series of patients (1 to 50) with a high incidence of positivity (100 to 56%). In other larger investigations, the detection of BoNT in sera was lower (about 20 to 35%). The clinical status of the patients who have been tested was not fully defined in all the reports. The detection of BoNT in the serum of patients with clinical symptoms of food-borne botulism is around 70% (56–82%) whatever the BoNT type, as indicated in the most documented publications [46,47,48,49,50,51]. BoNT is exceptionally reported in the serum of asymptomatic persons who ate the same contaminated foods as patients from a same outbreak of botulism. For example, BoNT/B was detected in the serum of two asymptomatic persons 87 days after ingestion of contaminated ham [43]. It was also reported BoNT/E and possibly BoNT/A in the serum of two asymptomatic cases 7 and 8 days, respectively, after consumption of toxic foods [44,45].
BoNT level in the patient’s sera depends on the ingested amount of toxin and the time elapsed between the ingestion of toxin and serum collection. In addition, variations of serum BoNT levels between patients from the same botulism outbreak have been observed. These individual variations likely result from different abilities in intestinal absorption of BoNT and/or clearance/elimination of the toxin, the mechanisms of which remain to be defined [28].
The levels of BoNT in naturally contaminated foods are very variable from <10 mouse lethal doses (MLD)/g to 280,000 MLD/g, as reported in the following publications [47,48,49,68,70,82]. The amount of ingested toxin by each patient can be estimated from the approximate portion which has been consumed, but it is usually unknown. Individual variations in the susceptibility to botulism are illustrated by the following example. An outbreak of botulism occurred in 2010 in Corsica in a family of 7 persons who ate a salad with home-canned green beans containing 2000 MLD/g BoNT/A2. According to the hostess, each person consumed an equivalent portion of salad. An 18-year-old patient developed a severe form of botulism and died of sudden respiratory arrest in less than 24 h. Three persons presented severe botulism and required mechanical ventilation for 37 to 78 days. Another guest showed a moderate form of botulism (diplopia, dysphagia, dysarthria), and the two remaining persons were asymptomatic. The levels of BoNT/A in the serum samples collected 1–2 days after the contaminated meal were 8 MLD/mL from the patient who died, 4 MLD/mL from two and 32–64 MLD/mL from one of the three patients who required intensive care, 4 MLD/mL from the patient with moderate botulism, and under the limit of detection from the two asymptomatic persons [49,83]. It is noteworthy that no obvious correlation was observed between the serum BoNT levels and the severity of symptoms [47]. However, high BoNT concentration in serum, notably BoNT/A, shortly (1–4 days) after ingestion of contaminated food is frequently associated with a severe form of botulism.
The levels of BoNT in the serum of patients are usually low. Most of the reported serum BoNT levels (>99%) range from 1 to 10 MLD/mL (Table 2). In some cases, the BoNT levels are <1 MLD/mL. Thus, injection of 1 mL of the patient’s serum into mice induces characteristic paralysis symptoms without lethality, which are neutralized by specific anti-BoNT antibodies. A small number of patients showed higher BoNT levels from 16 to 40 MLD/mL. Exceptional very high serum BoNT levels were reported (Table 2) such as 1800 MLD/mL of BoNT/A in a patient intoxicated with commercial carrot juice [68], and 400 MLD/mL of BoNT/F in a food-borne botulism case from an unknown origin [84,85].
The timeframe between ingestion of toxin and serum sampling is highly critical for successful BoNT detection. Indeed, the chances of obtaining positive results decrease when the time between the onset of symptoms and serum sampling increases. Most of the reported timeframes with positive BoNT detection in patient’s sera ranged from 1 to 4 days, whatever the botulism type (Table 3). In a series of 340 patient’s sera, BoNT was detected in >60% of serum samples collected within the two days after toxin ingestion, and positive results dropped to 13–28% in the sera obtained later [51]. However, long-lasting toxemia can be observed in certain patients. Most of the prolonged toxemia (more than 10 days) were observed in botulism type B (Table 2 and Table 3). Thereby, from 28 cases of botulism type B, the toxin was detected 2–3 days post-intoxication in the sera of 3 patients, 6 to 21 days in 23 patients, and 87 and 122 days in 2 other persons [43]. Sheth et al. [68] reported the presence of BoNT/A in the sera of two patients 12 and 25 days after illness onset. In another investigation, including 64 toxin positive sera, toxemia was detected in 15 patients 4 to 11 days after toxin ingestion [55] (Table 3). In an outbreak of botulism type A, the toxin was identified in the sera of 3 of 9 patients 10–12 days after toxin exposition [59]. Toxin detection in sera from patients with botulism type E or F was reported no more than 8 days after the disease onset (Table 2 and Table 3). Therefore, the chances of toxin detection in patient’s sera are maximal when serum sampling is as early as possible after toxin ingestion (ideally 1 to 4 days), but in some cases, very prolonged toxemia up to more than 120 days can be observed. The individual variations in the duration of botulism toxemia remain to be determined. L. Simpson suggests that BoNT elimination from the blood circulation most likely is achieved through hepatic uptake and biotransformation. The very low toxin levels in the blood, which are frequent in botulism type B, could not to be detected by the elimination system, thus accounting for prolonged toxemia [28].

3. Detection of BoNT in Sera of Patients with Intestinal Colonization

3.1. Infant Botulism

Botulism by intestinal colonization is characterized by the intestinal local production of BoNT, which is able to cross the intestinal barrier and to directly target the intestinal innervation. Efficient specific therapy for infant botulism consists of human botulism immunoglobulins (BabyBIG, BIG-IV) [39,86]. Experimentally, BoNT/A and BoNT/B administrated into the lumen of mouse intestinal loop crossed the epithelial cell barrier and specifically recognized neurons, mainly cholinergic neurons, in the intestinal submucosa and musculosa within 20–60 min [34,87,88]. Indeed, in addition to progressive weakness, a typical early symptom of infant botulism is constipation [18,19,21,89]. This suggests that synthesized BoNT in the intestine is, in part, eliminated with the stools, and another part is directly trapped by the enteric nervous system resulting in loss of intestinal motility and secretion. Thus, no remaining toxin or only a small fraction of remaining toxin is available for transport to the blood and lymph circulation. In contrast, in food-borne botulism, the ingestion of a BoNT bolus, likely in a larger amount than the toxin synthesized locally in the intestine, leads to a more abundant toxin proportion being absorbed and transported into the general circulation. Thereby, the presence of BoNT in the sera of infants with botulism has been less frequently reported than in sera from patients with food-borne botulism. Among 112 investigated serum samples from infant botulism, about 28% were toxin positive (Table 4), compared to the estimated 70% positivity in food-borne botulism (see above). In the large series of suspected infant botulism in the US, BoNT was identified in 9 sera of 67 infants (13%) with positive C. botulinum culture feces and none in 114 sera from infants with negative culture feces. In contrast, the toxin was recovered in the stools of most (87%) of the culture-positive infants [90]. Thus, the presence of neurotoxigenic Clostridium and toxin in feces seems to be the hallmark of infant botulism, while toxemia is less frequent. Therefore, laboratory confirmation of infant botulism mainly results from the detection of C. botulinum and BoNT in stool samples.
Most of the toxin positive sera from infant botulism are observed within 3–4 days after the symptom onset (Table 5), albeit neurotoxigenic Clostridium and toxin may be present in stools for up to several months [89,103,107,112,113,114]. In only a few cases, toxemia was recorded within a longer period, 7 to 27 days [92,93,113]. The toxin concentration in the sera from infant botulism is low (1 to 8 MLD/mL) and often at the limit of detection by the mouse bioassay [48,49,93], despite high BoNT levels in the intestinal content. For example, BoNT was not detected in the sera from 4 infants whose stools contained BoNT ranging from 2000 to 300,000 MLD/g [48,107]. This further argues for the weak transport efficiency of BoNT through the intestinal barrier and subsequent passage in the blood circulation [28,87] and supports that the largest part of toxin produced in the intestine during the clostridial growth is likely eliminated with the stools.

3.2. Botulism by Intestinal Colonization

More rarely, botulism by intestinal colonization occurs in infants above 1 year and in adults (adult intestinal toxemia botulism). Predisposing factors allowing spore germination, bacterial growth, and toxin production in the intestine include recent use of antibiotics, which can perturb the resident microbiota, previous gastric and/or intestinal surgery, and bowel anomalies such as Meckel’s diverticulum [22,95,115]. In a limited number of cases, the pathophysiology of botulism by intestinal colonization is similar to that of infant botulism. Of 12 cases reported in the literature with toxin investigation in the serum, BoNT was identified in 10 of them (Table 6). Most sera (9/12) were toxin positive when collected between 5 and 22 days after symptom onset. The longest timeframe was observed in a 27–year old man with botulism type B 47 days after the ingestion of suspected food. The onset of symptoms coincided with the appearance of toxin in the serum, suggesting a progressive C. botulinum growth and toxin production in the intestine [116].

4. Detection of BoNT in the Sera from Patients with Wound Botulism

Wound botulism is prevalent in certain countries such as California and UK (Table 7), and is mainly associated with risk factors, notably drug injection and more rarely accidental or surgically infected wounds. Local growth of C. botulinum in the anaerobic condition of infected wound leads to production of toxin, which diffuses through the extracellular space of the tissues to the local nerve endings and to the blood circulation. It is noteworthy that the clinical signs are identical to those of food-borne botulism but without gastrointestinal symptoms [24]. Of 335 patients with wound botulism reported in the literature, BoNT was identified in about 70% of them (Table 7). Toxin detection in the serum of patients with wound botulism is similar to that in food-borne botulism cases and higher than that in botulism by intestinal colonization. The time of collection of serum samples has not been reported in the cases of wound botulism. We can assume that serum sampling has been done shortly after the onset of symptoms corresponding to the maximum chances of toxin recovery in serum. The incubation period of wound botulism is estimated to 2–18 days [125,126]. In patients with late-onset of symptoms, 7–18 days after the initial wound contamination, a slow and progressive release of toxin with low amount in the blood circulation under the detection limit might account for the negative BoNT detection in serum.

5. Detection of Toxin in Sera of Patients with Neuropathic Symptoms Mimicking Botulism

Descending flaccid paralysis of botulism might be confused with neuropathic syndromes such as Guillain-Barré (GBS), Miller Fischer (MFS), Eaton-Lambert syndrome (ELS) or myasthenia [24]. Notably, some atypical cases of these neuropathies or myasthenia are difficult to distinguish clinically from botulism. GBS and MFS frequently mimic botulism [24,135,136,137]. GBS and MFS are immune neuropathies characterized by high titers of serum antibodies against gangliosides, in particular anti-GM1, anti-GD1a, anti-GD1b, anti-GT1b in GBS, and anti-GQ1b in MFS [138,139,140,141,142,143]. The laboratory confirmation of the differential diagnosis between botulism and autoimmune neuropathies is based on the identification of BoNT or antiganglioside antibodies in the patient’s sera, respectively. However, mouse bioassay can detect a toxicity that is not related to BoNT and which might lead to a false diagnostic of botulism. Indeed, toxicity of the patient’s serum by the mouse bioassay has been found in some cases of GBS and MFS (Table 8). Thus, intraperitoneal injection of serum from certain GBS patients into mouse induce respiratory distress and muscle paralysis similarly to BoNT administration [144]. Most often, the toxicity of serum from patients with GBS or MFS is not specifically neutralized by anti-botulinum toxin type antibodies leading to a conclusion of serum toxicity not related to BoNT [47,144,145]. In rare cases, BoNT/B or BoNT/A has been falsely identified in sera from MFS patients, but the presence of high level of anti-GQ1b antibodies or electromyography investigation supported the diagnosis of MFS [146,147]. The identification of BoNT toxinotype is done when only the mice injected with the patient’s serum sample incubated with the corresponding anti-BoNT toxinotype survive and all other mice die. However, if the patient’s serum’s toxicity is low ((1 LD50/mL), some mice can survive, leading to a false neutralization by a specific antiserum. It was observed that additional serum samples 5–6 days after a first toxin positive serum from GBS patients were no longer toxic [145]. Thus, serum toxicity from GBS and MFS patients seems to be low and for a short duration. The toxicity of the serum from patients with autoimmune neuropathy detected by the mouse bioassay is likely due to the autoantibodies against gangliosides. Indeed, antibodies against the gangliosides GM1, GD1a, GQ1b, or acetylcholine receptor impair acetylcholine release at neuromuscular junctions and mediate injury of motoneurons leading to muscle paralysis in mice [148,149,150,151,152,153,154]. Moreover, anti-GM1 antibodies induce flaccid paralysis in rabbit [155]. Monoclonal antibodies against GM1 and GD1a reduce the neurotransmitter release by decreasing the depolarization-induced calcium influx [156]. Anti-GQ1b and anti-GT1a block the transmission at neuromuscular junctions in a complement-dependent manner [151,157]. It is noteworthy that the gangliosides GD1a, GT1b, GQ1b are neuronal membrane targets of both BoNTs and autoantibodies [3,158], that might account for the similarity of symptoms between botulism and autoimmune neuropathies.

6. Alternative Methods of BoNT Detection in Serum

Since, in most cases, the BoNT level in the patient’s sera is low, a highly sensitive and specific method of toxin detection is required. The mouse bioassay is very sensitive, but it needs a laboratory with facility and authorization for animal handling. In addition, this in vivo assay might take up to 24–96 h, and it might interfere with non-related BoNT toxicity of sera from autoimmune neuropathies. Alternative in vitro methods of BoNT detection are based on immunological detection such as ELISA or other immunoassays, as well as on the BoNT enzymatic activity towards their specific SNARE protein substrates (SNAP25, VAMP, syntaxin) (Table 9) [42,161,162,163,164]. The sensitivity of the in vitro methods that have been developed with spiked or clinical human sera, is similar or lower than that of the mouse bioassay. Higher sensitivity than the mouse bioassay has been reported for some BoNT types, notably A and B (Table 8). However, the in vitro methods do not detect the full biological activity of BoNTs. Most of the in vitro methods use a two-step strategy, including a BoNT immunocapture with specific polyclonal or monoclonal antibodies most often directed against the Hc domain, followed by an endopeptidase assay with optimized SNARE peptide substrates. The cleavage products of the SNARE peptides are monitored either by immunological, fluorimetric, or mass spectrometry (MS) methods (Table 8). Immunological detection of the SNARE cleavage products benefits from the generation of neoepitopes in BoNT cleaved SNARE proteins. Thereby, specific antibodies recognize SNARE substrates cleaved by each BoNT type and not the native SNARE proteins (reviewed in [161,165]). The advantages of the in vitro methods are the rapidity (2.5 to 6 h) and the precise determination of the BoNT type, thus allowing accurate differentiation between botulism and autoimmune neuropathies as tested with clinical human sera [166,167] (Table 8). However, the in vitro assays require specific development for each BoNT type, while the mouse bioassay is sensitive to all BoNT types, and immunoassay detection is not specific of BoNT active forms.

7. Concluding Remarks

The paradigm of the physiopathology of botulism consists of BoNT absorption from the digestive tract/respiratory tract/wound, passage into the blood circulation, and then distribution to the target neurons [2,28]. The pharmacokinetics of BoNT has been investigated in experimental animals (mouse, rat) after intravenous or oral toxin administration. Orally administrated BoNT is detectable in the blood circulation after a latency period (5 h in mice receiving 1 or 4 μg BoNT/A1 complex by oral route) [169]. BoNT that reaches the blood circulation shows a biphasic decline: an initial and short phase with the maximum levels of BoNT, then a long phase with progressive BoNT decrease. The first phase is considered as the distribution phase of BoNT to the extracellular fluid compartments and the second slow phase as the elimination phase and clearance of BoNT from the blood circulation. BoNT is essentially free in the serum/plasma without binding to blood cells, only a small fraction (24%) of BoNT is bound to albumin. BoNT remains highly stable in the blood circulation. No proteolytic degradation, structural change, or loss of activity was observed. Thus, the blood circulation is considered as a “holding compartment”. BoNT elimination from the blood likely results from liver uptake and hepatic biotransformation [28,169,187,188,189]. BoNT in blood has been reported to be free toxin molecules (150,000 Da) non-associated to non-toxic molecules of botulinum complexes [119,190]. It is not clear whether the whole botulinum complexes enter the blood circulation and then dissociate or whether only free BoNT molecules have access to the blood compartment.
The pharmacokinetics of BoNT in humans has not been fully defined. It probably shows a similar pattern than in experimental animals, notably supporting the long persistence of BoNT in the serum of certain patients. BoNT detection in serum is the most direct way to confirm a diagnosis of botulism [41,42,191]. However, the presence of BoNT in the blood circulation depends on the time between serum sampling and the symptom onset, the initial amount of toxin involved in the disease, which has been ingested or produced locally, and probably of individual host factors related to toxin absorption and/or elimination which remain to be defined. BoNT levels in the patient’s sera is most commonly very low, thus requiring highly sensitive and specific methods of detection. In most cases, positive BoNT detection in serum is obtained when the serum sampling is performed within a few days after the disease onset. Based on the reported investigations, BoNT was detected in the serum of about 70% of food-borne botulism and wound botulism patients, and only in 28% of infant botulism. The number of investigations in older infants and adults with botulism by intestinal colonization was too small to be significant. BoNT produced locally in the intestine is likely eliminated in most part by the digestive tract. Only a BoNT fraction probably acts locally, leading to constipation that is a frequent early symptom of infant botulism, and a further smaller fraction enters the blood circulation. In contrast, the whole BoNT amount produced in a contaminated wound is presumed to be available for local activity and dissemination to the blood circulation, accounting for the higher prevalence of toxemia in wound botulism than in infant botulism.
Whether the mouse bioassay is still the most sensitive routine laboratory method to confirm clinical botulism by BoNT detection in serum, alternative in vitro methods such as based on the endopeptidase activity are promising. Their sensitivity is higher than that of the mouse bioassay for certain BoNT types, thus improving the diagnosis of botulism. The in vitro methods offer the advantage to identify more accurately the BoNT types and thus to better differentiate botulism from the autoimmune neuropathies and myasthenia.

Author Contributions

Conceptualization, M.R.P.; writing and editing, M.R.P.; review, C.R.-E. and E.L. All authors read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Conflicts of Interest

The authors declare no conflict of interest

References

  1. Dong, M.; Stenmark, P. The Structure and Classification of Botulinum Toxins. In Handbook of Experimental Pharmacology; Springer: Berlin/Heidelberg, Germany, 2019; pp. 1–23. [Google Scholar]
  2. Pirazzini, M.; Rossetto, O.; Eleopra, R.; Montecucco, C. Botulinum Neurotoxins: Biology, Pharmacology, and Toxicology. Pharm. Rev. 2017, 69, 200–235. [Google Scholar] [CrossRef] [PubMed]
  3. Poulain, B.; Lemichez, E.; Popoff, M.R. Neuronal selectivity of botulinum neurotoxins. Toxicon 2020, 178, 20–32. [Google Scholar] [CrossRef] [PubMed]
  4. Tehran, D.A.; Pirazzini, M. Novel botulinum neurotoxins: Exploring underneath the iceberg tip. Toxins 2018, 10, 190. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  5. Peck, M.W.; Smith, T.J.; Anniballi, F.; Austin, J.W.; Bano, L.; Bradshaw, M.; Cuervo, P.; Cheng, L.W.; Derman, Y.; Dorner, B.G.; et al. Historical perspectives and guidelines for botulinum neurotoxin subtype nomenclature. Toxins 2017, 9, 38. [Google Scholar] [CrossRef] [PubMed]
  6. Brunt, J.; van Vliet, A.H.M.; Stringer, S.C.; Carter, A.T.; Lindström, M.; Peck, M.W. Pan-Genomic Analysis of Clostridium botulinum group II (Non-Proteolytic C. botulinum) associated with foodborne botulism and isolated from the environment. Toxins 2020, 12, 306. [Google Scholar] [CrossRef]
  7. Rasetti-Escargueil, C.; Lemichez, E.; Popoff, M.R. Variability of botulinum toxins: Challenges and opportunities for the future. Toxins 2018, 10, 374. [Google Scholar] [CrossRef] [Green Version]
  8. Zornetta, I.; Azarnia Tehran, D.; Arrigoni, G.; Anniballi, F.; Bano, L.; Leka, O.; Zanotti, G.; Binz, T.; Montecucco, C. The first non Clostridial botulinum-like toxin cleaves VAMP within the juxtamembrane domain. Sci. Rep. 2016, 6, 30257. [Google Scholar] [CrossRef]
  9. Brunt, J.; Carter, A.T.; Stringer, S.C.; Peck, M.W. Identification of a novel botulinum neurotoxin gene cluster in Enterococcus. FEBS Lett. 2018, 592, 310–317. [Google Scholar] [CrossRef] [Green Version]
  10. Zhang, S.; Lebreton, F.; Mansfield, M.J.; Miyashita, S.I.; Zhang, J.; Schwartzman, J.A.; Tao, L.; Masuyer, G.; Martinez-Carranza, M.; Stenmark, P.; et al. Identification of a botulinum neurotoxin-like toxin in a commensal strain of Enterococcus faecium. Cell Host Microbe 2018, 23, 169–176. [Google Scholar] [CrossRef] [Green Version]
  11. Mansfield, M.J.; Wentz, T.G.; Zhang, S.; Lee, E.J.; Dong, M.; Sharma, S.K.; Doxey, A.C. Bioinformatic discovery of a toxin family in Chryseobacterium piperi with sequence similarity to botulinum neurotoxins. Sci. Rep. 2019, 9, 1634. [Google Scholar] [CrossRef]
  12. Wentz, T.G.; Muruvanda, T.; Lomonaco, S.; Thirunavukkarasu, N.; Hoffmann, M.; Allard, M.W.; Hodge, D.R.; Pillai, S.P.; Hammack, T.S.; Brown, E.W.; et al. Closed genome sequence of Chryseobacterium piperi strain CTM(T)/ATCC BAA-1782, a gram-negative bacterium with clostridial neurotoxin-like coding sequences. Genome Announc. 2017, 5. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  13. Contreras, E.; Masuyer, G.; Qureshi, N.; Chawla, S.; Dhillon, H.S.; Lee, H.L.; Chen, J.; Stenmark, P.; Gill, S.S. A neurotoxin that specifically targets Anopheles mosquitoes. Nat. Commun. 2019, 10, 2869. [Google Scholar] [CrossRef] [PubMed]
  14. Dong, M.; Masuyer, G.; Stenmark, P. Botulinum and tetanus neurotoxins. Annu. Rev. Biochem. 2019, 88, 811–837. [Google Scholar] [CrossRef] [PubMed]
  15. Rummel, A. Two Feet on the membrane: Uptake of clostridial neurotoxins. Curr. Top. Microbiol. Immunol. 2017, 406, 1–37. [Google Scholar] [CrossRef] [PubMed]
  16. Peck, M.W. Clostridium botulinum and the safety of minimally heated, chilled foods: An emerging issue? J. Appl. Microbiol. 2006, 101, 556–570. [Google Scholar] [CrossRef]
  17. Peck, M.W.; Stringer, S.C.; Carter, A.T. Clostridium botulinum in the post-genomic era. Food Microbiol. 2011, 28, 183–191. [Google Scholar] [CrossRef]
  18. Brook, I. Infant botulism. J. Perinatol. 2007, 27, 175–180. [Google Scholar] [CrossRef] [Green Version]
  19. Rosow, L.K.; Strober, J.B. Infant botulism: Review and clinical update. Pediatric Neurol. 2015, 52, 487–492. [Google Scholar] [CrossRef]
  20. Shirey, T.B.; Dykes, J.K.; Lúquez, C.; Maslanka, S.E.; Raphael, B.H. Characterizing the fecal microbiota of infants with botulism. Microbiome 2015, 3, 54. [Google Scholar] [CrossRef] [Green Version]
  21. Fox, C.K.; Keet, C.A.; Strober, J.B. Recent advances in infant botulism. Pediatr. Neurol. 2005, 32, 149–154. [Google Scholar] [CrossRef]
  22. Harris, R.A.; Anniballi, F.; Austin, J.W. Adult intestinal toxemia botulism. Toxins 2020, 12, 81. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  23. Franz, D.R.; Pitt, L.M.; Clayton, M.A.; Hanes, M.A.; Rose, K.J. Efficacy of prophylactic and therapeutic administration of antitoxin for inhalation botulism. In Botulinum and Tetanus Neurotoxins; DasGupta, B.R., Ed.; Plenum Press: New York, NY, USA, 1993; pp. 473–476. [Google Scholar]
  24. Sobel, J. Botulism. Clin. Infect. Dis. 2005, 41, 1167–1173. [Google Scholar] [CrossRef] [PubMed]
  25. Popoff, M.R. Botulinum Toxins, Diversity, Mode of Action, Epidemiology of Botulism in France. In Botulinum Toxin; Nikolay, S., Ed.; IntechOpen: London, UK, 2018; pp. 1–28. [Google Scholar] [CrossRef] [Green Version]
  26. Ghasemi, M.; Norouzi, R.; Salari, M.; Asadi, B. Iatrogenic botulism after the therapeutic use of botulinum toxin-A: A case report and review of the literature. Clin. Neuropharmacol. 2012, 35, 254–257. [Google Scholar] [CrossRef] [PubMed]
  27. Floresta, G.; Patamia, V.; Gentile, D.; Molteni, F.; Santamato, A.; Rescifina, A.; Vecchio, M. Repurposing of FDA-approved drugs for treating iatrogenic botulism: A paired 3d-qsar/docking approach(†). ChemMedChem 2020, 15, 256–262. [Google Scholar] [CrossRef] [PubMed]
  28. Simpson, L. The life history of a botulinum toxin molecule. Toxicon 2013, 68, 40–59. [Google Scholar] [CrossRef] [PubMed]
  29. Rummel, A. The long journey of botulinum neurotoxins into the synapse. Toxicon 2015, 107, 9–24. [Google Scholar] [CrossRef] [PubMed]
  30. Benefield, D.A.; Dessain, S.K.; Shine, N.; Ohi, M.D.; Lacy, D.B. Molecular assembly of botulinum neurotoxin progenitor complexes. Proc. Natl. Acad. Sci. USA 2013, 110, 5630–5635. [Google Scholar] [CrossRef] [Green Version]
  31. Poulain, B.; Molgo, J.; Popoff, M.R. Clostridial Neurotoxins: From the Cellular and Molecular Mode of Action to Their Therapeutic Use. In The Comprehensive Sourcebook of Bacterial Protein Toxins, 4th ed.; Alouf, J., Ladant, D., Popoff, M.R., Eds.; Elsevier: Amsterdam, The Netherlands, 2015; pp. 287–336. [Google Scholar]
  32. Gu, S.; Jin, R. Assembly and function of the botulinum neurotoxin progenitor complex. Curr. Top. Microbiol. Immunol. 2013, 364, 21–44. [Google Scholar] [CrossRef]
  33. Gu, S.; Rumpel, S.; Zhou, J.; Strotmeier, J.; Bigalke, H.; Perry, K.; Shoemaker, C.B.; Rummel, A.; Jin, R. Botulinum neurotoxin is shielded by NTNHA in an interlocked complex. Science 2012, 335, 977–981. [Google Scholar] [CrossRef] [Green Version]
  34. Connan, C.; Varela-Chavez, C.; Mazuet, C.; Molgo, J.; Haustant, G.M.; Disson, O.; Lecuit, M.; Vandewalle, A.; Popoff, M.R. Translocation and dissemination to target neurons of botulinum neurotoxin type B in the mouse intestinal wall. Cell. Microbiol. 2016, 18, 282–301. [Google Scholar] [CrossRef]
  35. Fujinaga, Y.; Popoff, M.R. Translocation and dissemination of botulinum neurotoxin from the intestinal tract. Toxicon 2018, 147, 13–18. [Google Scholar] [CrossRef] [PubMed]
  36. Fujinaga, Y.; Sugawara, Y.; Matsumura, T. Uptake of botulinum neurotoxin in the intestine. Curr. Top. Microbiol. Immunol. 2013, 364, 45–59. [Google Scholar] [PubMed]
  37. Arnon, S.S.; Schechter, R.; Maslanka, S.E.; Jewell, N.P.; Hatheway, C.L. Human botulism immune globulin for the treatment of infant botulism. N. Engl. J. Med. 2006, 354, 462–471. [Google Scholar] [CrossRef] [PubMed]
  38. Rasetti-Escargueil, C.; Popoff, M.R. Antibodies and vaccines against botulinum toxins: Available measures and novel approaches. Toxins 2019, 11, 528. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  39. Payne, J.R.; Khouri, J.M.; Jewell, N.P.; Arnon, S.S. Efficacy of human botulism immune globulin for the treatment of infant botulism: The first 12 years post licensure. J. Pediatr. 2018, 193, 172–177. [Google Scholar] [CrossRef]
  40. Sobel, J. Diagnosis and treatment of botulism: A century later, clinical suspicion remains the cornerstone. Clin. Infect. Dis. 2009, 48, 1674–1675. [Google Scholar] [CrossRef] [Green Version]
  41. Lonati, D.; Schicchi, A.; Crevani, M.; Buscaglia, E.; Scaravaggi, G.; Maida, F.; Cirronis, M.; Petrolini, V.M.; Locatelli, C.A. Foodborne botulism: Clinical diagnosis and medical treatment. Toxins 2020, 12, 509. [Google Scholar] [CrossRef]
  42. Lindström, M.; Korkeala, H. Laboratory diagnosis of botulism. Clin. Microbiol. Rev. 2006, 19, 298–314. [Google Scholar] [CrossRef] [Green Version]
  43. Sebald, M.; Saimot, G. Toxémie botulique. Intérêt de sa mise en évidence dans le diagnostic du botulisme humain de type B. Ann. Microbiol. 1973, 124, 61–69. [Google Scholar]
  44. Koenig, M.G.; Spickard, A.; Cardella, M.A.; Rogers, D.E. Clinical and laboratory observations on type e botulism in man. Medical 1964, 43, 517–545. [Google Scholar] [CrossRef]
  45. Koenig, M.G.; Drutz, D.J.; Mushlin, A.I.; Schaffner, W.; Rogers, D.E. Type B botulism in man. Am. J. Med. 1967, 42, 208–219. [Google Scholar] [CrossRef]
  46. Sebald, M.; Saimot, G. Le diagnostic biologique du botulisme. Med. Mal. Inf. 1973, 3, 83–85. [Google Scholar] [CrossRef]
  47. Mazuet, C.; Bouvet, P.; King, L.A.; Popoff, M.R. Le botulisme humain en France, 2007–2009. Bull. Epidemiol. Hebd. 2011, 6, 49–53. [Google Scholar]
  48. Mazuet, C.; Jourdan-Da Silva, N.; Legeay, C.; Sautereau, J.; Popoff, M.R. Le botulisme humain en France, 2013-2016. Bull. Epidémiol. Hebd. 2018, 3, 46–54. [Google Scholar]
  49. Mazuet, C.; King, L.A.; Bouvet, P.; Legeay, C.; Sautereau, J.; Popoff, M.R. Le botulisme humain en France, 2010-2012. Bull. Epidemiol. Hebd. 2014, 6, 106–114. [Google Scholar]
  50. Roblot, P.; Roblot, F.; Fauchère, J.L.; Devilleger, A.; Maréchaud, R.; Breux, J.P.; Grollier, G.; Becq-Giraudon, B. Retrospective study of 108 cases of botulism in Poitiers France. J. Med. Microbiol. 1994, 40, 379–384. [Google Scholar] [CrossRef] [PubMed]
  51. Woodruff, B.A.; Griffin, P.M.; McCroskey, L.M.; Smart, J.F.; Wainwright, R.B.; Bryant, R.G.; Hutwagner, L.C.; Hatheway, C.L. Clinical and laboratory comparison of botulism from toxin types A, B, and E in the United States, 1975–1988. J. Infect. Dis. 1992, 166, 1281–1286. [Google Scholar] [CrossRef]
  52. Blake, P.A.; Horwitz, M.A.; Hopkins, L.; Lombard, G.L.; McCroan, J.E.; Prucha, J.C.; Merson, M.H. Type A botulism from commercially canned beef stew. South. Med. J. 1977, 70, 5–7. [Google Scholar] [CrossRef]
  53. Mann, J.M.; Hatheway, C.L.; Gardiner, T.M. Laboratory diagnosis in a large outbreak of type A botulism: Confirmation of the value of coproexamination. Am. J. Epidemiol. 1982, 115, 598–605. [Google Scholar] [CrossRef]
  54. Takahashi, M.; Tokumaru, Y.; Yamamoto, M.; Shimizu, T.; Kondo, H.; Ishihara, N.; Okubo, T.; Sato, T.; Sakaguchi, G. A case report on human type B botulism. Jpn. J. Med. Sci. Biol. 1986, 39, 29–33. [Google Scholar] [CrossRef] [Green Version]
  55. Fagan, R.P.; McLaughlin, J.B.; Middaugh, J.P. Persistence of botulinum toxin in patients’ serum: Alaska, 1959–2007. J. Infect. Dis. 2009, 199, 1029–1031. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  56. McCroskey, L.M.; Hatheway, C.L.; Woodruff, B.A.; Greenberg, J.A.; Jurgenson, P. Type F botulism due to neurotoxigenic Clostridium baratii from an unknown source in an adult. J. Clin. Microbiol. 1991, 29, 2618–2620. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  57. Townes, J.M.; Cieslak, P.R.; Hatheway, C.L.; Solomon, H.M.; Holloway, J.T.; Baker, M.P.; Keller, C.F.; McCroskey, L.M.; Griffin, P.M. An outbreak of type A botulism associated with a commercial cheese sauce. Ann. Intern. Med. 1996, 125, 558–563. [Google Scholar] [CrossRef] [PubMed]
  58. Korkeala, H.; Stengel, G.; Hyytia, E.; Vogelsang, B.; Bohl, A.; Wihlman, H.; Pakkala, P.; Hielm, S. Type E botulism associated with vacuum-packaged hot-smoked whitefish. Int. J. Food Microbiol. 1998, 43, 1–5. [Google Scholar] [CrossRef]
  59. Villar, R.G.; Shapiro, R.L.; Busto, S.; Riva-Posse, C.; Verdejo, G.; Farace, M.I.; Rosetti, F.; San Juan, J.A.; Julia, C.M.; Becher, J.; et al. Outbreak of type A botulism and development of a botulism surveillance and antitoxin release system in Argentina. JAMA 1999, 281, 1334–1338. [Google Scholar] [CrossRef]
  60. Lindstrôm, M.; Hielm, S.; Nevas, M.; Tuisku, S.; Korkeala, H. Proteolytic Clostridium botulinum type B in the gastric content of a patient with type E botulism due to whitefish eggs. Foodborne Pathog. Dis. 2004, 1, 53–57. [Google Scholar] [CrossRef]
  61. Ouagari, Z.; Chakib, A.; Sodqi, M.; Marih, L.; Marhoum Filali, K.; Benslama, A.; Idrissi, L.; Moutawakkil, S.; Himmich, H. Botulism in Casablanca. (11 cases). Bull. Soc. Pathol. Exot. 2002, 95, 272–275. [Google Scholar]
  62. Bielec, D.; Semczuk, G.; Lis, J.; Firych, J.; Modrzewska, R.; Janowski, R. Clinical and epidemiological analysis of patients with botulism hospitalized at the Department of Infectious Disease, Medical University of Lublin in 1990-2000. Przegl. Epidemiol. 2002, 56, 435–442. [Google Scholar]
  63. Leclair, D.; Fung, J.; Isaac-Renton, J.L.; Proulx, J.F.; May-Hadford, J.; Ellis, A.; Ashton, E.; Bekal, S.; Farber, J.M.; Blanchfield, B.; et al. Foodborne botulism in Canada, 1985–2005. Emerg. Infect. Dis. 2013, 19, 961–968. [Google Scholar] [CrossRef]
  64. Harvey, S.M.; Sturgeon, J.; Dassey, D.E. Botulism due to Clostridium baratii type F toxin. J. Clin. Microbiol. 2002, 40, 2260–2262. [Google Scholar] [CrossRef] [Green Version]
  65. Merz, B.; Bigalke, H.; Stoll, G.; Naumann, M. Botulism type B presenting as pure autonomic dysfunction. Clin. Auton. Res. 2003, 13, 337–338. [Google Scholar] [CrossRef] [PubMed]
  66. Sobel, J.; Dill, T.; Kirkpatrick, C.L.; Riek, L.; Luedtke, P.; Damrow, T.A. Clinical recovery and circulating botulinum toxin type F in adult patient. Emerg. Infect. Dis. 2009, 15, 969–971. [Google Scholar] [CrossRef] [PubMed]
  67. Akdeniz, H.; Buzgan, T.; Tekin, M.; Karsen, H.; Karahocagil, M.K. An outbreak of botulism in a family in Eastern Anatolia associated with eating süzme yoghurt buried under soil. Scand. J. Infect. Dis. 2007, 39, 108–114. [Google Scholar] [CrossRef] [PubMed]
  68. Sheth, A.N.; Wiersma, P.; Atrubin, D.; Dubey, V.; Zink, D.; Skinner, G.; Doerr, F.; Juliao, P.; Gonzalez, G.; Burnett, C.; et al. International outbreak of severe botulism with prolonged toxemia caused by commercial carrot juice. Clin. Infect. Dis. 2008, 47, 1245–1251. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  69. Lindstrôm, M.; Vuorela, M.; Hinderink, K.; Korkeala, H.; Dahlsten, E.; Raahenmaa, M.; Kuusi, M. Botulism associated with vacuum-packed smoked whitefish in Finland, June–July 2006. Eurosurveillance 2006, 11, E060720–E060723. [Google Scholar] [CrossRef]
  70. Juliao, P.C.; Maslanka, S.; Dykes, J.; Gaul, L.; Bagdure, S.; Granzow-Kibiger, L.; Salehi, E.; Zink, D.; Neligan, R.P.; Barton-Behravesh, C.; et al. National outbreak of type a foodborne botulism associated with a widely distributed commercially canned hot dog chili sauce. Clin. Infect. Dis. 2013, 56, 376–382. [Google Scholar] [CrossRef] [Green Version]
  71. Hannett, G.E.; Schaffzin, J.K.; Davis, S.W.; Fage, M.P.; Schoonmaker-Bopp, D.; Dumas, N.B.; Musser, K.A.; Egan, C. Two cases of adult botulism caused by botulinum neurotoxin producing Clostridium baratii. Anaerobe 2014, 30, 178–180. [Google Scholar] [CrossRef]
  72. Sobel, J.; Malavet, M.; John, S. Outbreak of clinically mild botulism type E illness from home-salted fish in patients presenting with predominantly gastrointestinal symptoms. Clin. Infect. Dis. 2007, 45, e14–e16. [Google Scholar] [CrossRef] [Green Version]
  73. Rowlands, R.E.; Ristori, C.A.; Lopes, G.I.; de Paula, A.M.; Sakuma, H.; Grigaliunas, R.; Lopreato Filho, R.; Gelli, D.S.; Eduardo, M.B.; Jakabi, M. Botulism in Brazil, 2000–2008: Epidemiology, clinical findings and laboratorial diagnosis. Rev. Inst. Med. Trop. São Paulo 2010, 52, 183–186. [Google Scholar] [CrossRef] [Green Version]
  74. Jalava, K.; Selby, K.; Pihlajasaari, A.; Kolho, E.; Dahlsten, E.; Forss, N.; Backlund, T.; Korkeala, H.; Honkanen-Buzalski, T.; Hulkko, T.; et al. Two cases of food-borne botulism in Finland caused by conserved olives, October 2011. Eurosurveillance 2011, 16, 20034. [Google Scholar] [CrossRef] [Green Version]
  75. Browning, L.; Prempeh, H.; Little, C.; Houston, C.; Grant, K.; Cowden, J. An outbreak of food-borne botulism in Scotland, United Kingdom, November 2011. Eurosurveillance 2011, 16, 20036. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  76. Momose, Y.; Asakura, H.; Kitamura, M.; Okada, Y.; Ueda, Y.; Hanabara, Y.; Sakamoto, T.; Matsumura, T.; Iwaki, M.; Kato, H.; et al. Food-borne botulism in Japan in March 2012. Int. J. Infect. Dis. 2014, 24, 20–22. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  77. Potulska-Chromik, A.; Zakrzewska-Pniewska, B.; Szmidt-Salkowska, E.; Lewandowski, J.; Sinski, M.; Przyjalkowski, W.; Kostera-Pruszczyk, A. Long lasting dysautonomia due to botulinum toxin B poisoning: Clinical-laboratory follow up and difficulties in initial diagnosis. BMC Res. Notes 2013, 6, 438. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  78. McCarty, C.L.; Angelo, K.; Beer, K.D.; Cibulskas-White, K.; Quinn, K.; de Fijter, S.; Bokanyi, R.; St Germain, E.; Baransi, K.; Barlow, K.; et al. Large Outbreak of Botulism Associated with a Church Potluck Meal--Ohio, 2015. Morb. Mortal. Wkly. Rep. 2015, 64, 802–803. [Google Scholar] [CrossRef] [PubMed]
  79. Proverbio, M.R.; Lamba, M.; Rossi, A.; Siani, P. Early diagnosis and treatment in a child with foodborne botulism. Anaerobe 2016, 39, 189–192. [Google Scholar] [CrossRef] [PubMed]
  80. Anniballi, F.; Auricchio, B.; Fiore, A.; Lonati, D.; Locatelli, C.A.; Lista, F.; Fillo, S.; Mandarino, G.; De Medici, D. Botulism in Italy, 1986 to 2015. Eurosurveillance 2017, 22. [Google Scholar] [CrossRef] [Green Version]
  81. Hellmich, D.; Wartenberg, K.E.; Zierz, S.; Mueller, T.J. Foodborne botulism due to ingestion of home-canned green beans: Two case reports. J. Med. Case Rep. 2018, 12, 1. [Google Scholar] [CrossRef] [Green Version]
  82. Carlier, J.P.; Espié, E.; Popoff, M.R. Le botulisme en France, 2003–2006. Bull. Epidemiol. Hebd. 2007, 31–32, 281–284. [Google Scholar]
  83. Oriot, C.; D’Aranda, E.; Castanier, M.; Glaizal, M.; Galy, C.; Faivre, A.; Poisnel, E.; Truong, T.K.; Mercury, P.; Hayek-Lanthois, M.; et al. One collective case of type A foodborne botulism in Corsica. Clin. Toxicol. 2011, 49, 752–754. [Google Scholar] [CrossRef]
  84. Mazuet, C.; Sautereau, J.; Legeay, C.; Bouchier, C.; Bouvet, P.; Jourdan da Silva, N.; Castor, C.; Popoff, M.R. Characterization of the first Clostridium baratii strain responsible for an outbreak of botulism type F in France. Clin. Microbiol. Case Rep. 2015, 1, 1–4. [Google Scholar]
  85. Mazuet, C.; Legeay, C.; Sautereau, J.; Bouchier, C.; Criscuolo, A.; Bouvet, P.; Trehard, H.; Jourdan Da Silva, N.; Popoff, M. Characterization of Clostridium baratii Type F strains responsible for an outbreak of botulism linked to beef meat consumption in France. PLoS Curr. Outbreaks 2017. [Google Scholar] [CrossRef]
  86. Khouri, J.M.; Payne, J.R.; Arnon, S.S. More clinical mimics of infant botulism. J. Pediatr. 2018, 193, 178–182. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  87. Connan, C.; Popoff, M.R. Uptake of clostridial neurotoxins into cells and dissemination. Curr. Top. Microbiol. Immunol. 2017, 406, 39–78. [Google Scholar] [CrossRef] [PubMed]
  88. Couesnon, A.; Molgo, J.; Connan, C.; Popoff, M.R. Preferential entry of botulinum neurotoxin A Hc domain trhough intestinal crypt cells and targeting to cholinergic neurons of the mouse intestine. PLoS Pathog. 2012, 8, e1002583. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  89. Arnon, S.S. Infant botulism. Annu. Rev. Med. 1980, 31, 541–560. [Google Scholar] [CrossRef] [PubMed]
  90. Hatheway, C.; McCroskey, L.M. Examination of feces and serum for diagnosis of infant botulism in 336 patients. J. Clin. Microbiol. 1987, 25, 2334–2338. [Google Scholar] [CrossRef] [Green Version]
  91. Paton, J.C.; Lawrence, A.J.; Manson, J.I. Quantitation of Clostridium botulinum organisms and toxin in the feces of an infant with botulism. J. Clin. Microbiol. 1982, 15, 1–4. [Google Scholar] [CrossRef] [Green Version]
  92. Paton, J.C.; Lawrence, A.J.; Steven, I.M. Quantities of Clostridium botulinum organisms and toxin in feces and presence of Clostridium botulinum toxin in the serum of an infant with botulism. J. Clin. Microbiol. 1983, 17, 13–15. [Google Scholar] [CrossRef] [Green Version]
  93. Takahashi, M.; Noda, H.; Takeshita, S.; Fujiwara, T.; Nakanoin, H.; Mizunoya, T.; Sakaguchi, G. Attempts to quantity Clostridium botulinum type A toxin and antitoxin in serum of two cases of infant botulism in Japan. Jpn. J. Med. Sci. Biol. 1990, 43, 233–237. [Google Scholar] [CrossRef] [Green Version]
  94. Fenicia, L.; Anniballi, F. Infant botulism. Ann. Ist. Super. Sanita 2009, 45, 134–146. [Google Scholar]
  95. Fenicia, L.; Anniballi, F.; Aureli, P. Intestinal toxemia botulism in Italy, 1984–2005. Eur. J. Clin. Microbiol. Infect. Dis. 2007, 26, 385–394. [Google Scholar] [CrossRef] [PubMed]
  96. Grant, B.D.; Donaldson, J.G. Pathways and mechanisms of endocytic recycling. Nat. Rev. Mol. Cell Biol. 2009, 10, 597–608. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  97. Balslev, T.; Ostergaard, E.; Madsen, I.K.; Wandall, D.A. Infant botulism. The first culture-confirmed Danish case. Neuropediatrics 1997, 28, 287–288. [Google Scholar] [CrossRef] [PubMed]
  98. Brett, M.M.; McLauchlin, J.; Harris, A.; O’Brien, S.; Black, N.; Forsyth, R.J.; Roberts, D.; Bolton, F.J. A case of infant botulism with a possible link to infant formula milk powder: Evidence for the presence of more than one strain of Clostridium botulinum in clinical specimens and food. J. Med. Microbiol. 2005, 54, 769–776. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  99. Nevas, M.; Hielm, S.; Lindstrôm, M.; Horn, H.; Koivulehto, K.; Korkeala, H. High prevalence of Clostridium botulinum types A and B in honey samples detected by polymerase chain reaction. Int. J. Food Microbiol. 2002, 72, 45–52. [Google Scholar] [CrossRef]
  100. Luquez, C.; Dykes, J.K.; Yu, P.A.; Raphael, B.H.; Maslanka, S.E. First report worldwide of an infant botulism case due to Clostridium botulinum type E. J. Clin. Microbiol. 2010, 48, 326–328. [Google Scholar] [CrossRef] [Green Version]
  101. Paerregaard, A.; Angen, O.; Lisby, M.; Mølbak, K.; Clausen, M.E.; Christensen, J.J. Denmark: Botulism in an infant or infant botulism? Eurosurveillance 2008, 13, 19072. [Google Scholar]
  102. Saraiva, M.; Campos Cunha, I.; Costa Bonito, C.; Pena, C.; Toscano, M.M.; Teixeira Lopes, T.; Sousa, I.; Calhau, M.A. First case of infant botulism in Portugal. Food Control 2012, 26, 79–80. [Google Scholar] [CrossRef]
  103. Derman, Y.; Korkeala, H.; Salo, E.; Lonnqvist, T.; Saxen, H.; Lindström, M. Infant botulism with prolonged faecal excretion of botulinum neurotoxin and Clostridium botulinum for 7 months. Epidemiol. Infect. 2014, 142, 335–339. [Google Scholar] [CrossRef]
  104. Bernardor, J.; Neveu, J.; Haas, H.; Pitelet, G.; Popoff, M.R.; Mazuet, C.; Berard, E.; Boulay, C.; Chabrol, B. Infant botulism: Two case reports and electroneuromyogram findings. Arch. Pediatr. 2018, 25, 340–343. [Google Scholar]
  105. Hoarau, G.; Pelloux, I.; Gayot, A.; Wroblewski, I.; Popoff, M.R.; Mazuet, C.; Maurin, M.; Croize, J. Two cases of type A infant botulism in Grenoble, France: No honey for infants. Eur. J. Pediatr. 2012, 171, 589–591. [Google Scholar] [CrossRef]
  106. King, L.A.; Popoff, M.R.; Mazuet, C.; Espié, E.; Vaillant, V.; de Valk, H. Infant botulism in France, 1991-2009. Arch. Pediatr. 2010, 17, 1288–1292. [Google Scholar] [CrossRef] [PubMed]
  107. Mazuet, C.; Yoon, E.J.; Boyer, S.; Pignier, S.; Blanc, T.; Doehring, I.; Meziane-Cherif, D.; Dumant-Forest, C.; Sautereau, J.; Legeay, C.; et al. A penicillin- and metronidazole-resistant Clostridium botulinum strain responsible for an infant botulism case. Clin. Microbiol. Infect. 2016, 22, 644.e7–644.e12. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  108. Paricio, C.; Bey, K.J.; Teyssier, G.; Ughetto, A.; Ros, A.; Rayet, I.; Lavocat, M.P. Botulism in a neonate. Arch. Pediatr. 2006, 13, 146–148. [Google Scholar] [CrossRef]
  109. Patural, H.; Goffaux, P.; Paricio, C.; Emeriaud, G.; Teyssier, G.; Barthelemy, J.C.; Pichot, V.; Roche, F. Infant botulism intoxication and autonomic nervous system dysfunction. Anaerobe 2009, 15, 197–200. [Google Scholar] [CrossRef] [PubMed]
  110. Breinbjerg, A.; Rittig, S.; Kamperis, K. Persistent neurogenic bladder dysfunction due to infantile botulism. BMJ Case Rep. 2014, 2014, 202077. [Google Scholar] [CrossRef] [Green Version]
  111. Spini, R.G.; Ferraris, V.; Glasman, M.P.; Orofino, G.; Casanovas, A.; Debaisi, G. Intussusception in infant with diagnostic botulism: A case report. Arch. Argent. Pediatr. 2015, 113, e286–e289. [Google Scholar] [CrossRef]
  112. Fenicia, L.; Da Dalt, L.; Anniballi, F.; Franciosa, G.; Zanconato, S.; Aureli, P. A case if infant botulism due to neurotoxigenic Clostridium butyricum type E associated with Clostridium difficile colitis. Eur. J. Clin. Microbiol. Infect. Dis. 2002, 21, 736–738. [Google Scholar] [CrossRef]
  113. Grant, K.A.; Nwarfor, I.; Mpamugo, O.; Mithani, V.; Lister, P.; Dixon, G.; Nixon, G.; Planche, T.; Courtney, M.; Morgan, J.; et al. Report of two unlinked cases of infant botulism in the UK in October 2007. J. Med. Microbiol. 2009, 58, 1601–1606. [Google Scholar] [CrossRef]
  114. Wilcke, B.W., Jr.; Midura, T.F.; Arnon, S.S. Quantitative evidence of intestinal colonization by Clostridium botulinum in four cases of infant botulism. J. Infect. Dis. 1980, 141, 419–423. [Google Scholar] [CrossRef]
  115. Griffin, P.M.; Hatheway, C.L.; Rosenbaum, R.B.; Sokolow, R. Endogenous antibody production to botulinum toxin in an adult with intestinal colonization botulism and underlying Crohn’s disease. J. Infect. Dis. 1997, 175, 633–637. [Google Scholar] [CrossRef] [Green Version]
  116. McCroskey, L.M.; Hatheway, C.L. Laboratory findings in four cases of adult botulism suggest colonization of the intestinal tract. J. Clin. Microbiol. 1988, 26, 1052–1054. [Google Scholar] [CrossRef] [Green Version]
  117. Chia, J.K.; Clark, J.B.; Ryan, C.A.; Pollack, M. Botulism in an adult associated with food-borne intestinal infection with Clostridium Botulinum. N. Engl. J. Med. 1986, 315, 239–241. [Google Scholar] [CrossRef]
  118. Fenicia, L.; Franciosa, G.; Pourshaban, M.; Aureli, P. Intestinal toxemia botulism in two young people, caused by Clostridium butyricum Type E. Clin. Infect. Dis. 1999, 29, 381–387. [Google Scholar] [CrossRef] [Green Version]
  119. Kobayashi, H.; Fujisawa, K.; Saito, Y.; Kamijo, M.; Oshima, S.; Kubo, M.; Eto, Y.; Monma, C.; Kitamura, M. A botulism case of a 12-year-old girl caused by intestinal colonization of Clostridium botulinum type Ab. Jpn. J. Infect. Dis. 2003, 56, 73–74. [Google Scholar]
  120. Sheppard, Y.D.; Middleton, D.; Whitfield, Y.; Tyndel, F.; Haider, S.; Spiegelman, J.; Swartz, R.H.; Nelder, M.P.; Baker, S.L.; Landry, L.; et al. Intestinal toxemia botulism in 3 adults, Ontario, Canada, 2006–2008. Emerg. Infect. Dis. 2012, 18, 1–6. [Google Scholar] [CrossRef]
  121. Freund, B.; Hayes, L.; Rivera-Lara, L.; Sumner, C.; Chaudhry, V.; Chatham-Stephens, K.; Benedict, K.; Kalb, S.; Blythe, D.; Brooks, R.; et al. Adult intestinal colonization botulism mimicking brain death. Muscle Nerve 2017, 56, E27–E28. [Google Scholar] [CrossRef]
  122. Halai, U.A.; Terashita, D.; Kim, M.; Green, N.; Kalb, S.R.; Chatham-Stephens, K.; Balter, S. Notes from the Field: Intestinal colonization and possible iatrogenic botulism in mouse bioassay-negative serum specimens—Los Angeles County, California, November 2017. Morb. Mortal. Wkly. Rep. 2018, 67, 1221–1222. [Google Scholar] [CrossRef]
  123. Parameswaran, L.; Rao, A.; Chastain, K.; Ackelsburg, J.; Adams, E.; Jackson, B.; Voigt, L.P.; Chen, X.; Boulad, F.; Taur, Y. A Case of adult intestinal toxemia botulism during prolonged hospitalization in an allogeneic hematopoietic cell transplant recipient. Clin. Infect. Dis. 2017, 66, S99–S102. [Google Scholar] [CrossRef] [Green Version]
  124. Guru, P.K.; Becker, T.L.; Stephens, A.; Cannistraro, R.J.; Eidelman, B.H.; Hata, D.J.; Brumble, L. Adult intestinal botulism: A rare presentation in an immunocompromised patient with short bowel syndrome. Mayo Clin. Proc. Innov. Qual Outcomes 2018, 2, 291–296. [Google Scholar] [CrossRef] [Green Version]
  125. Brett, M.M.; Hallas, G.; Mpamugo, O. Wound botulism in the UK and Ireland. J. Med. Microbiol. 2004, 53, 555–561. [Google Scholar] [CrossRef] [Green Version]
  126. Werner, S.B.; Passaro, D.; McGee, J.; Schechter, R.; Vugia, D.J. Wound botulism in California, 1951-1998: Recent epidemic in heroin injectors. Clin. Infect. Dis. 2000, 31, 1018–1024. [Google Scholar] [CrossRef] [PubMed]
  127. Yuan, J.; Inami, G.; Mohle-Boetani, J.; Vugia, D.J. Recurrent wound botulism among injection drug users in California. Clin. Infect. Dis. 2011, 52, 862–866. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  128. Wheeler, C.; Inami, G.; Mohle-Boetani, J.; Vugia, D. Sensitivity of mouse bioassay in clinical wound botulism. Clin. Infect. Dis. 2009, 48, 1669–1673. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  129. Akbulut, D.; Dennis, J.; Gent, M.; Grant, K.A.; Hope, V.; Ohai, C.; McLauchlin, J.; Mithani, V.; Mpamugo, O.; Ncube, F.; et al. Wound botulism in injectors of drugs: Upsurge in cases in England during 2004. Eurosurveillance 2005, 10, 172–174. [Google Scholar] [CrossRef] [PubMed]
  130. Kalka-Moll, W.M.; Aurbach, U.; Schaumann, R.; Schwarz, R.; Seifert, H. Wound botulism in injection drug users. Emerg. Infect. Dis. 2007, 13, 942–943. [Google Scholar] [CrossRef]
  131. Artin, I.; Bjorkman, P.; Cronqvist, J.; Radstrom, P.; Holst, E. First case of type E wound botulism diagnosed using real-time PCR. J. Clin. Microbiol. 2007, 45, 3589–3594. [Google Scholar] [CrossRef] [Green Version]
  132. Nystrom, S.C.; Wells, E.V.; Pokharna, H.S.; Johnson, L.E.; Najjar, M.A.; Mamou, F.M.; Rudrik, J.T.; Miller, C.E.; Boulton, M.L. Botulism toxemia following laparoscopic appendectomy. Clin. Infect. Dis. 2012, 54, e32–e34. [Google Scholar] [CrossRef] [Green Version]
  133. Schulte, M.; Hamsen, U.; Schildhauer, T.A.; Ramczykowski, T. Effective and rapid treatment of wound botulism, a case report. BMC Surg. 2017, 17, 103. [Google Scholar] [CrossRef] [Green Version]
  134. Cook, P.A.; Mishler, A.; Quan, D.; Parrish-Garcia, A. Wound botulism caused by Clostridium subterminale after a heroin injection. Infect. Dis. Rep. 2018, 10, 7654. [Google Scholar] [CrossRef] [Green Version]
  135. Wakerley, B.R.; Yuki, N. Mimics and chameleons in Guillain-Barré and Miller Fisher syndromes. Pract. Neurol. 2015, 15, 90–99. [Google Scholar] [CrossRef]
  136. Legast, G.M.; Lascano, A.M.; Gschwind, M.; Schnider, A.; Nicastro, N. Guillain-Barré and Miller Fisher overlap syndrome mimicking alimentary botulism. J. Clin. Neurol. 2017, 13, 442–443. [Google Scholar] [CrossRef] [PubMed]
  137. Susuki, K.; Takahashi, H.; Yuki, N.; Ohsawa, H.; Hirata, K.; Sakata, I. Guillain-Barre syndrome mimicking botulism. J. Neurol. 2001, 248, 720–721. [Google Scholar] [CrossRef] [PubMed]
  138. Chiba, A.; Kusunoki, S.; Obata, H.; Machinami, R.; Kanazawa, I. Serum anti-GQ1b IgG antibody is associated with ophthalmoplegia in Miller Fisher syndrome and Guillain-Barré syndrome: Clinical and immunohistochemical studies. Neurological 1993, 43, 1911–1917. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  139. Kusunoki, S.; Kaida, K. Antibodies against ganglioside complexes in Guillain-Barré syndrome and related disorders. J. Neurochem. 2011, 116, 828–832. [Google Scholar] [CrossRef] [PubMed]
  140. Wanleenuwat, P.; Iwanowski, P.; Kozubski, W. Antiganglioside antibodies in neurological diseases. J. Neurol. Sci. 2020, 408, 116576. [Google Scholar] [CrossRef] [PubMed]
  141. Cutillo, G.; Saariaho, A.H.; Meri, S. Physiology of gangliosides and the role of antiganglioside antibodies in human diseases. Cell. Mol. Immunol. 2020, 17, 313–322. [Google Scholar] [CrossRef]
  142. Willison, H.J.; Yuki, N. Peripheral neuropathies and anti-glycolipid antibodies. Brain 2002, 125, 2591–2625. [Google Scholar] [CrossRef]
  143. Ogawa, G.; Kaida, K.; Kuwahara, M.; Kimura, F.; Kamakura, K.; Kusunoki, S. An antibody to the GM1/GalNAc-GD1a complex correlates with development of pure motor Guillain-Barré syndrome with reversible conduction failure. J. Neuroimmunol. 2013, 254, 141–145. [Google Scholar] [CrossRef]
  144. Van den Berg, L.H.; Oey, P.L.; Wokke, J.H.; Veldman, H.; Wieneke, G.H.; Notermans, S.H. Features of the Guillain-Barré syndrome in mice following intraperitoneal injection of patient serum. J. Neurol. Sci. 1994, 127, 103–106. [Google Scholar] [CrossRef]
  145. Notermans, S.H.; Wokke, J.H.; van den Berg, L.H. Botulism and Guillain-Barré syndrome. Lancet 1992, 340, 303. [Google Scholar] [CrossRef]
  146. Chang, V.H.; Robinson, L.R. Serum positive botulism with neuropathic features. Arch. Phys. Med. Rehabil. 2000, 81, 122–126. [Google Scholar] [CrossRef]
  147. Zeylikman, Y.; Shah, V.; Shah, U.; Mirsen, T.R.; Campellone, J.V. False-Positive Serum Botulism Bioassay in Miller-Fisher Syndrome. J. Clin. Neuromuscul. Dis. 2015, 17, 27–29. [Google Scholar] [CrossRef] [PubMed]
  148. Vernino, S.; Ermilov, L.G.; Sha, L.; Szurszewski, J.H.; Low, P.A.; Lennon, V.A. Passive transfer of autoimmune autonomic neuropathy to mice. J. Neurosci. 2004, 24, 7037–7042. [Google Scholar] [CrossRef] [PubMed]
  149. Goodfellow, J.A.; Bowes, T.; Sheikh, K.; Odaka, M.; Halstead, S.K.; Humphreys, P.D.; Wagner, E.R.; Yuki, N.; Furukawa, K.; Furukawa, K.; et al. Overexpression of GD1a ganglioside sensitizes motor nerve terminals to anti-GD1a antibody-mediated injury in a model of acute motor axonal neuropathy. J. Neurosci. 2005, 25, 1620–1628. [Google Scholar] [CrossRef] [PubMed]
  150. Plomp, J.J.; Willison, H.J. Pathophysiological actions of neuropathy-related anti-ganglioside antibodies at the neuromuscular junction. J. Physiol. 2009, 587, 3979–3999. [Google Scholar] [CrossRef] [PubMed]
  151. Goodyear, C.S.; O’Hanlon, G.M.; Plomp, J.J.; Wagner, E.R.; Morrison, I.; Veitch, J.; Cochrane, L.; Bullens, R.W.; Molenaar, P.C.; Conner, J.; et al. Monoclonal antibodies raised against Guillain-Barré syndrome-associated Campylobacter jejuni lipopolysaccharides react with neuronal gangliosides and paralyze muscle-nerve preparations. J. Clin. Investig. 1999, 104, 697–708. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  152. Taguchi, K.; Ren, J.; Utsunomiya, I.; Aoyagi, H.; Fujita, N.; Ariga, T.; Miyatake, T.; Yoshino, H. Neurophysiological and immunohistochemical studies on Guillain-Barre syndrome with IgG anti-GalNAc-GD1a antibodies-effects on neuromuscular transmission. J. Neurol. Sci. 2004, 225, 91–98. [Google Scholar] [CrossRef]
  153. Taguchi, K.; Utsunomiya, I.; Ren, J.; Yoshida, N.; Aoyagi, H.; Nakatani, Y.; Ariga, T.; Usuki, S.; Yu, R.K.; Miyatake, T. Effect of rabbit anti-asialo-GM1 (GA1) polyclonal antibodies on neuromuscular transmission and acetylcholine-induced action potentials: Neurophysiological and immunohistochemical studies. NeuroChem. Res. 2004, 29, 953–960. [Google Scholar] [CrossRef]
  154. Asthana, P.; Vong, J.S.; Kumar, G.; Chang, R.C.; Zhang, G.; Sheikh, K.A.; Ma, C.H. Dissecting the role of anti-ganglioside antibodies in Guillain-Barré Syndrome: An animal model approach. Mol. Neurobiol. 2016, 53, 4981–4991. [Google Scholar] [CrossRef]
  155. Yuki, N.; Yamada, M.; Koga, M.; Odaka, M.; Susuki, K.; Tagawa, Y.; Ueda, S.; Kasama, T.; Ohnishi, A.; Hayashi, S.; et al. Animal model of axonal Guillain-Barré syndrome induced by sensitization with GM1 ganglioside. Ann. Neurol. 2001, 49, 712–720. [Google Scholar] [CrossRef]
  156. Buchwald, B.; Zhang, G.; Vogt-Eisele, A.K.; Zhang, W.; Ahangari, R.; Griffin, J.W.; Hatt, H.; Toyka, K.V.; Sheikh, K.A. Anti-ganglioside antibodies alter presynaptic release and calcium influx. Neurobiol. Dis. 2007, 28, 113–121. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  157. Bullens, R.W.; O’Hanlon, G.M.; Goodyear, C.S.; Molenaar, P.C.; Conner, J.; Willison, H.J.; Plomp, J.J. Anti-GQ1b antibodies and evoked acetylcholine release at mouse motor endplates. Muscle Nerve 2000, 23, 1035–1043. [Google Scholar] [CrossRef]
  158. Bullens, R.W.; O’Hanlon, G.M.; Wagner, E.; Molenaar, P.C.; Furukawa, K.; Furukawa, K.; Plomp, J.J.; Willison, H.J. Complex gangliosides at the neuromuscular junction are membrane receptors for autoantibodies and botulinum neurotoxin but redundant for normal synaptic function. J. Neurosci. 2002, 22, 6876–6884. [Google Scholar] [CrossRef] [PubMed]
  159. Dorr, J.; Dieste, F.J.; Klaasen van Husen, D.; Zipp, F.; Vogel, H.P. A case of recurrent Miller Fisher syndrome mimicking botulism. Neurol. Sci. 2006, 27, 424–425. [Google Scholar] [CrossRef] [PubMed]
  160. Hargis, M.J.; Howdeshell, M. Miller-Fisher Syndrome With Botulism Toxin Ingestion: The Ultimate Descending Paralysis. J. Clin. Neuromuscul. Dis. 2018, 19, 146. [Google Scholar] [CrossRef]
  161. Hobbs, R.J.; Thomas, C.A.; Halliwell, J.; Gwenin, C.D. Rapid Detection of Botulinum Neurotoxins-A Review. Toxins 2019, 11, 418. [Google Scholar] [CrossRef] [Green Version]
  162. Nepal, M.R.; Jeong, T.C. Alternative methods for testing botulinum toxin: Current status and future perspectives. Biomol. Ther. 2020, 28, 302–310. [Google Scholar] [CrossRef]
  163. Perry, M.J.; Centurioni, D.A.; Davis, S.W.; Hannett, G.E.; Musser, K.A.; Egan, C.T. Implementing the Bruker MALDI biotyper in the public health laboratory for C. botulinum neurotoxin detection. Toxins 2017, 9, 94. [Google Scholar] [CrossRef] [Green Version]
  164. Stern, D.; von Berg, L.; Dorner, M.B.; Dorner, B.G. Replacing the mouse bioassay for diagnostics and potency testing of botulinum neurotoxins—Rogress and challenges. Berl. Munch. Tierarztl. Wochenschr. 2018, 131, 375–394. [Google Scholar]
  165. Kalb, S.R.; Boyer, A.E.; Barr, J.R. Mass spectrometric detection of bacterial protein toxins and their enzymatic activity. Toxins 2015, 7, 3497–3511. [Google Scholar] [CrossRef] [Green Version]
  166. Ferracci, G.; Marconi, S.; Mazuet, C.; Jover, E.; Blanchard, M.P.; Seagar, M.; Popoff, M.; Leveque, C. A label-free biosensor assay for botulinum neurotoxin B in food and human serum. Anal. Biochem. 2011, 410, 281–288. [Google Scholar] [CrossRef] [PubMed]
  167. Leveque, C.; Ferracci, G.; Maulet, Y.; Mazuet, C.; Popoff, M.; Seagar, M.; El Far, O. Direct biosensor detection of botulinum neurotoxin endopeptidase activity in sera from patients with type A botulism. Biosens. Bioelectron. 2014, 57, 207–212. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  168. Bagramyan, K.; Barash, J.R.; Arnon, S.S.; Kalkum, M. Attomolar detection of botulinum toxin type A in complex biological matrices. PLoS ONE 2008, 3, e2041. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  169. Bagramyan, K.; Kaplan, B.E.; Cheng, L.W.; Strotmeier, J.; Rummel, A.; Kalkum, M. Substrates and controls for the quantitative detection of active botulinum neurotoxin in protease-containing samples. Anal. Chem. 2013, 85, 5569–5576. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  170. Von Berg, L.; Stern, D.; Pauly, D.; Mahrhold, S.; Weisemann, J.; Jentsch, L.; Hansbauer, E.M.; Muller, C.; Avondet, M.A.; Rummel, A.; et al. Functional detection of botulinum neurotoxin serotypes A to F by monoclonal neoepitope-specific antibodies and suspension array technology. Sci. Rep. 2019, 9, 5531. [Google Scholar] [CrossRef] [Green Version]
  171. Kalb, S.R.; Moura, H.; Boyer, A.E.; McWilliams, L.G.; Pirkle, J.L.; Barr, J.R. The use of Endopep-MS for the detection of botulinum toxins A, B, E, and F in serum and stool samples. Anal. Biochem. 2006, 351, 84–92. [Google Scholar] [CrossRef]
  172. Wang, D.; Baudys, J.; Krilich, J.; Smith, T.J.; Barr, J.R.; Kalb, S.R. A two-stage multiplex method for quantitative analysis of botulinum neurotoxins type a, B, e, and f by maldi-tof mass spectrometry. Anal. Chem. 2014, 86, 10847–10854. [Google Scholar] [CrossRef]
  173. Wang, D.; Krilich, J.; Baudys, J.; Barr, J.R.; Kalb, S.R. Optimization of peptide substrates for botulinum neurotoxin E improves detection sensitivity in the Endopep-MS assay. Anal. Biochem. 2015, 468, 15–21. [Google Scholar] [CrossRef] [Green Version]
  174. Wang, D.; Baudys, J.; Hoyt, K.M.; Barr, J.R.; Kalb, S.R. Further optimization of peptide substrate enhanced assay performance for BoNT/A detection by MALDI-TOF mass spectrometry. Anal. Bioanal. Chem. 2017, 409, 4779–4786. [Google Scholar] [CrossRef] [Green Version]
  175. Ruge, D.R.; Dunning, F.M.; Piazza, T.M.; Molles, B.E.; Adler, M.; Zeytin, F.N.; Tucker, W.C. Detection of six serotypes of botulinum neurotoxin using fluorogenic reporters. Anal. Biochem. 2011, 411, 200–209. [Google Scholar] [CrossRef] [Green Version]
  176. Dunning, F.M.; Ruge, D.R.; Piazza, T.M.; Stanker, L.H.; Zeytin, F.N.; Tucker, W.C. Detection of botulinum neurotoxin serotype A, B, and F proteolytic activity in complex matrices with picomolar to femtomolar sensitivity. Appl. Environ. Microbiol. 2012, 78, 7687–7697. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  177. Rosen, O.; Feldberg, L.; Dor, E.; Gura, S.; Zichel, R. Optimization of SNAP-25-derived peptide substrate for improved detection of botulinum A in the Endopep-MS assay. Anal. Biochem. 2017, 528, 34–37. [Google Scholar] [CrossRef] [PubMed]
  178. Rosen, O.; Feldberg, L.; Yamin, T.S.; Dor, E.; Barnea, A.; Weissberg, A.; Zichel, R. Development of a multiplex Endopep-MS assay for simultaneous detection of botulinum toxins A, B and E. Sci. Rep. 2017, 7, 14859. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  179. Parks, B.A.; Shearer, J.D.; Baudys, J.; Kalb, S.R.; Sanford, D.C.; Pirkle, J.L.; Barr, J.R. Quantification of botulinum neurotoxin serotypes A and B from serum using mass spectrometry. Anal. Chem. 2011, 83, 9047–9053. [Google Scholar] [CrossRef]
  180. Björnstad, K.; Tevell Åberg, A.; Kalb, S.R.; Wang, D.; Barr, J.R.; Bondesson, U.; Hedeland, M. Validation of the Endopep-MS method for qualitative detection of active botulinum neurotoxins in human and chicken serum. Anal. Bioanal. Chem. 2014, 406, 7149–7161. [Google Scholar] [CrossRef] [Green Version]
  181. Wang, D.; Krilich, J.; Baudys, J.; Barr, J.R.; Kalb, S.R. Enhanced detection of type C botulinum neurotoxin by the Endopep-MS assay through optimization of peptide substrates. Bioorgan. Med. Chem. 2015, 23, 3667–3673. [Google Scholar] [CrossRef] [Green Version]
  182. Babrak, L.; Lin, A.; Stanker, L.H.; McGarvey, J.; Hnasko, R. Rapid microfluidic assay for the detection of botulinum neurotoxin in animal sera. Toxins 2016, 8, 13. [Google Scholar] [CrossRef]
  183. Jones, R.G.; Marks, J.D. Use of a new functional dual coating (FDC) assay to measure low toxin levels in serum and food samples following an outbreak of human botulism. J. Med. Microbiol. 2013, 62, 828–835. [Google Scholar] [CrossRef] [Green Version]
  184. Liu, Y.Y.; Rigsby, P.; Sesardic, D.; Marks, J.D.; Jones, R.G. A functional dual-coated (FDC) microtiter plate method to replace the botulinum toxin LD50 test. Anal. Biochem. 2012, 425, 28–35. [Google Scholar] [CrossRef]
  185. Mazuet, C.; Ezan, E.; Volland, H.; Popoff, M.R.; Becher, F. Toxin detection in patients’ sera by mass spectrometry during two outbreaks of type A Botulism in France. J. Clin. Microbiol. 2012, 50, 4091–4094. [Google Scholar] [CrossRef] [Green Version]
  186. Weisemann, J.; Krez, N.; Fiebig, U.; Worbs, S.; Skiba, M.; Endermann, T.; Dorner, M.B.; Bergstrom, T.; Munoz, A.; Zegers, I.; et al. Generation and characterization of six recombinant botulinum neurotoxins as reference material to serve in an international proficiency test. Toxins 2015, 7, 5035–5054. [Google Scholar] [CrossRef] [Green Version]
  187. Al-Saleem, F.H.; Ancharski, D.M.; Ravichandran, E.; Joshi, S.G.; Singh, A.K.; Gong, Y.; Simpson, L.L. The role of systemic handling in the pathophysiologic actions of botulinum toxin. J. Pharm. Exp. 2008, 326, 856–863. [Google Scholar] [CrossRef] [Green Version]
  188. Ravichandran, E.; Gong, Y.; Al Saleem, F.H.; Ancharski, D.M.; Joshi, S.G.; Simpson, L.L. An initial assessment of the systemic pharmacokinetics of botulinum toxin. J. Pharm. Exp. 2006, 318, 1343–1351. [Google Scholar] [CrossRef] [PubMed]
  189. Cheng, L.W.; Stanker, L.H.; Henderson, T.D., II; Lou, J.; Marks, J.D. Antibody protection against botulinum neurotoxin intoxication in mice. Infect. Immun. 2009, 77, 4305–4313. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  190. Kitamura, M.; Sakaguchi, S.; Sakaguchi, G. Significance of 12S toxin of Clostridium botulinum type E. J. Bacteriol. 1969, 98, 1173–1178. [Google Scholar] [CrossRef] [Green Version]
  191. Cai, S.; Singh, B.R.; Sharma, S. Botulism diagnostics: From clinical symptoms to in vitro assays. Crit. Rev. Microbiol. 2007, 33, 109–125. [Google Scholar] [CrossRef]
Table
Table 1. Detection of BoNT in the sera of patients with food-borne botulism.
Table 1. Detection of BoNT in the sera of patients with food-borne botulism.
CountryYearNumber of Patient’s SeraToxin-Positive Sera 1BoNT TypeReferences
ABEFNon-Identified
USA1905–19622020193 7[44]
USA196365 5 [44]
USA196544 4 [45]
France1970–19733928 28 [43]
USA1975–198834012671/10912/10926/109 [51]
South Africa1977211 [52]
USA1978181111 [53]
Japan198411 1 [54]
France1966–19905028 28 [50]
Alaska1959–2007180641/15 211/15 23/15 2 [55]
USA199111 1 3 [56]
USA1993522 [57]
Finland199711 1 [58]
Argentina1998933 [59]
Finland199911 1 [60]
Morocco1999116 6 [61]
Poland1990–20003224 231 [62]
Canada1985–200521274(8%) 4(6%) 4(86%) 4 [63]
USA200111 1 3 [64]
Germany200310 5 [65]
USA200511 1 3 [66]
Turkey2005922 [67]
USA2006655 [68]
Finland200611 1 [69]
USA2007822 [70]
USA200722 2 3 [71]
USA200750 6 [72]
Brazil2000–2008451784 7 5[73]
France2007–200934325261 [47]
France2010–2012514120182 1[49]
Finland201122 2 [74]
UK2011222 [75]
Japan2012222 [76]
Poland201311 1 [77]
USA2015191919 [78]
Italy201511 1 [79]
Italy1986–2015275561145 [80]
France2013–20165644733 4 3 [48]
Germany2018222 [81]
Total 1456633
1 Toxin detection in patient’s sera by mouse bioassay, 2 BoNT typing was reported for only 15 serum samples, 3 C. baratii producing BoNT/F detected in stool or food, 4 Distribution of the botulism types during this period, 5 BoNT/B detected in remaining ham, 6 BoNT/E detected in remnant salted fish, 7 Detection of BoNT types A and B.
Table
Table 2. BoNT concentration (mouse lethal dose (MLD)/mL) in the sera of patients with food-borne botulism.
Table 2. BoNT concentration (mouse lethal dose (MLD)/mL) in the sera of patients with food-borne botulism.
CountryYearNumber of Patient’s SeraDays from Symptom Onset to Serum SamplingBoNT TypeBoNT Concentration (MLD/mL)References
USA196351–10E1–10[44]
France 11970–19731120B1[43]
173–20 (most 8–15)B1–10
121B4–40
Japan198414B1–2[54]
Finland19971 E6[58]
Finland199911E1[60]
Turkey20059 A<1–2[67]
USA20061 (patient A)1A>20[68]
1 (patient B)5A>200
71800
8>200
12>200
France2007–20161 A80[47,48,49]
2 A32–40
2 A8–16
35 A1–8
6 B16–40
67 B1–8
14 B<1
1 E8
2 E1
Finland201113B<1[74]
France201412F (C. baratii)400[84,85]
121–2
France20153 F (C. baratii)1-4[85]
1 BoNT concentration determined in 19 sera of 28 toxin positive sera.
Table
Table 3. Detection of botulinum toxin in sera of patients with food-borne botulism and time between the symptom onset and serum sampling.
Table 3. Detection of botulinum toxin in sera of patients with food-borne botulism and time between the symptom onset and serum sampling.
CountryYearDays from Symptom Onset to Serum SamplingToxin Positive-SeraBoNT TypeAdditional DetectionReferences
France1970–1973 122B [43]
6–1816
20–212
872
1201
USA1975–19880–360–70% 2A, B, E [51]
≥ 413–28% 2
Japan198441B [54]
Alaska1959–20074–1111B [55]
5–73E
41A
USA199141F 3neg at day 14[56]
Argentina199810–123A [59]
Finland199911E [60]
USA200121F 3neg at day 6[64]
USA20051 and 81F 3 [66]
USA200613A [68]
5, 7, 8, 121 (patient B)A
251Aneg at day 41
Finland200611E [69]
USA200732 4A [70]
4, 9, 18, 25neg 6 4
USA20073, 62F 3 [71]
USA20071, 3neg 5mild botulism E [72]
Finland201132B [74]
Japan201222A [76]
Italy201511B [79]
Germany201832A [81]
1 Dates of serum sampling known for 23 of the 28 toxin positive sera, 2 From a total of 87 serum samples for which date specimen was obtained, 3 C. baratii F, 4 Sera from 8 patients of the same outbreak, negative detection of BoNT in 6 sera. neg, negative.
Table
Table 4. Detection of BoNT in the serum of infants with botulism.
Table 4. Detection of BoNT in the serum of infants with botulism.
CountryYearNumber of Infant’s SeraToxin-Positive SeraBoNT TypeReferences
ABE
USA1975–198767981 [90]
Australia198110 1 1 [91]
Australia1982111 [92]
Japan1986–1987222 [93]
Italy1984–2008122 11 2[94,95]
UK1994–20074422 [96]
Denmark1997111 [97]
UK200110 1 1 [98]
Finland200210 1 1 [99]
USA200711 1[100]
Denmark200811 3 [101]
Portugal200910 1 1 [102]
Finland2010111 [103]
France2004–201616954 [48,49,104,105,106,107,108,109].
Denmark2013111 [110]
Argentina2014111 [111]
Total 11231
1C. botulinum B in feces, 2 C. butyricum E in feces, 3 Botulinum toxin type not identified.
Table
Table 5. Detection of botulinum toxin in sera of infant botulism according to the time between the symptom onset and serum sampling.
Table 5. Detection of botulinum toxin in sera of infant botulism according to the time between the symptom onset and serum sampling.
CountryYearPatient Age at the Onset of Symptoms
m, months; d, days		Days from Symptom Onset to Serum SamplingBoNT in SerumC. botulinum in FecesReferences
Australia19814 m4negC. botulinum B[91]
Australia19827 d3AC. botulinum A[92]
10AC. botulinum A
16negC. botulinum A
Japan198679 d22negC. botulinum A[93]
198740 d7AC. botulinum A
24AC. botulinum A
26AC. botulinum A
27AC. botulinum A
46negC. botulinum A
UK20078 m15AC. botulinum A[113]
UK20015 m10negC. botulinum B[98]
USA20079 d5negC. botulinum E[100]
Denmark20084.5 m14BoNT 1 [101]
Finland20103 m3AC. botulinum A[103]
15negC. botulinum A
Denmark20135 m4AC. botulinum A[110]
France20132 m28negC. botulinum A[107]
48negC. botulinum A
63negC. botulinum A
Argentina20144 m3AC. botulinum A[111]
1 Botulinum toxin type not identified, neg, negative.
Table
Table 6. Detection of BoNT in the serum of patients with botulism by intestinal colonization.
Table 6. Detection of BoNT in the serum of patients with botulism by intestinal colonization.
CountryYearPatient AgeDays from Symptom Onset to Serum SamplingBoNT in SerumBoNT in FecesFecesReferences
USA1978–1985elderly22ndBC. botulinum B[116]
30BnegC. botulinum B
37negndnd
332AndC. botulinum A
2747BnegC. botulinum B
3714AAC. botulinum A
21negnegC. botulinum A
USA19853711AAC. botulinum A[117]
13AndC. botulinum A
USA1988673AAnd[115]
5AAC. botulinum A
~20negndC. botulinum A
Italy199495negndC. butyricum E[118]
Italy19975630AnegC. botulinum A[95]
Japan2003122AAC. botulinum Ab[119]
Canada20075013AndC. botulinum A[120]
France201110 Ind 1ndC. butyricum E[49]
USA20174312AndC. botulinum A[121]
USA2017elderly16A 1And[122]
USA20172778negAC. botulinum A[123]
USA2018665AAnd[124]
1 Toxin type undetermined. 2 Detection of BoNT/A negative by mouse bioassay and positive by MALDI-TOF MS. nd, not done; neg, negative.
Table
Table 7. Detection of BoNT in the serum of patients with wound botulism.
Table 7. Detection of BoNT in the serum of patients with wound botulism.
CountryYearNumber of Patient’s SeraToxin-Positive SeraBoNT TypeReferences
ABEind
California1951–199812211484%12% 4%[126]
California1993–20063722202 [127]
California2005–20077350 [128]
Italy1986–201565 5 [80]
UK2000–20048833~90%~10% [125,129]
Germany200542 2 [130]
Sweden200610 1 1 [131]
France200811 1 [47]
USA2011111 [132]
Germany201611 1 [133]
USA2017111 2 [134]
Total 335230
1C. botulinum E detected in the wound by real time PCR, 2 C. subterminale identified in wound culture.
Table
Table 8. Detection of botulinum toxin in sera of patients with neuropathic symptoms mimicking botulism.
Table 8. Detection of botulinum toxin in sera of patients with neuropathic symptoms mimicking botulism.
CountryYearNumber of Patient’s SeraToxin Positive SeraAdditional InvestigationSymptomsReferences
The Netherlands1988–199254 1 GBS[145]
The Netherlands199461 1CSF protein: 125 mg/dL, electromyographyGBS[144]
USA19981BoNT/BCSF protein: 42 mg/dL, electrodiagnosticpossible concurrent botulism and MFS[146]
Germany20041negstrongly elevated IgG/IgM anti-GQ1bMFS mimicking botulism[159]
France2007–20093824 1 GBS/MFS[47] and unpublished data
USA20151BoNT/Astrongly elevated Ig anti-GQ1b
CSF protein: 56 mg/dL
negative detection of BoNT and C. botulinum in feces		MFS[147]
USA20171negstrongly elevated IgG/IgM anti-GQ1b
BoNT in recently ingested canned food		possible concurrent MFS and food-borne botulism[160]
Switzerland20171negelevated IgG anti-GM1, -GD1a, -GD1batypical GBS/MFS mimicking botulism[136]
1 Toxicity in sera not neutralized by specific anti-botulinum toxin type antibodies. GBS, Guillain Barré syndrome; MFS, Miller-Fisher syndrome; neg, negative.
Table
Table 9. In vitro methods of BoNT detection in human serum.
Table 9. In vitro methods of BoNT detection in human serum.
BoNT TypeMethodSerum SamplesSensitivityReferences
AAssay with a large immunosorbent surface area (ALISSA)spiked human serumAttomolar
(4 × 10−5 MLD/mL) a		[168,169]
A, B, C, D, E, FEndopeptidase assay and monoclonal neoepitope antibodiesspiked human serumA: 0.211 MLD50/mL
B: 0.147 MLD50/mL
C: 2.056 MLD50/mL
D: 0.18 MLD50/mL
E: 0.84 MLD50/mL
F: 0.005 MLD50/mL		[170]
A, B, E, FEndopep-MSspiked human serumA: 20 MLD/mL
B, F: 1 MLD/mL
E: 0.2 MLD/mL		[171]
A, B, E, FEndopep-MALDI-TOF-MSspiked human serumA, B, F: 0.1 MLD/mL
E: 0.02 MLD/mL		[172,173,174]
A, B, FEndopeptidase assay with fluorogenic substratesspiked human serumA, F: 1 pM
(A: 40 MLD/mL
F: 3 MLD/mL) a
B: 10 pM
(180 MLD/mL) a		[175,176]
AEndopep-MSspiked human serum1 MLD/mL[177,178]
A, BEndopep-MSspiked macaque serum1 MLD/mL[179]
A, B, D/C, E, FEndopep-MSspiked human, chicken seraA, B, F: 1 MLD/mL
D/C: 2 MLD/mL
E: 10 MLD/mL
C: 100 MLD/mL		[180]
CEndopep-MSspiked human serum0.5–1 MLD/mL[181]
AMicrofluidic double sandwich immunoassayspiked human serum30 pg/mL
(8 MLD/mL) a		[182]
AFunctional dual coating assay (BoNT immunocapture and endopeptidase assay with detection by neoepitope antibodies)clinical serum samples1 MLD/mL[183,184]
AEndopeptidase assay and detection with neoepitope antibodies and surface plasmon resonanceclinical serum samples<1 MLD/mL
negative in ELS and CIDP patient’s sera		[167]
AEndopep-MSclinical serum samples~1 MLD/mL[185]
BEndopeptidase assay and detection with neoepitope antibodies and surface plasmon resonanceclinical serum samples0.1–0.01 MLD/mL
negative in ELS and GBS patient’s sera		[166]
Endopep-MS, endopeptidase assay and mass spectrometry; ELS, Eaton-Lambert syndrome; CIDP, chronic inflammatory demyelinating neuropathy; GBS, Guillain-Barré syndrome. a The equivalence in MLD/mL has been determined according to [186].
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Share and Cite

MDPI and ACS Style

Rasetti-Escargueil, C.; Lemichez, E.; Popoff, M.R. Toxemia in Human Naturally Acquired Botulism. Toxins 2020, 12, 716. https://doi.org/10.3390/toxins12110716

AMA Style

Rasetti-Escargueil C, Lemichez E, Popoff MR. Toxemia in Human Naturally Acquired Botulism. Toxins. 2020; 12(11):716. https://doi.org/10.3390/toxins12110716

Chicago/Turabian Style

Rasetti-Escargueil, Christine, Emmanuel Lemichez, and Michel R. Popoff. 2020. "Toxemia in Human Naturally Acquired Botulism" Toxins 12, no. 11: 716. https://doi.org/10.3390/toxins12110716

APA Style

Rasetti-Escargueil, C., Lemichez, E., & Popoff, M. R. (2020). Toxemia in Human Naturally Acquired Botulism. Toxins, 12(11), 716. https://doi.org/10.3390/toxins12110716

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop