Last data update: Apr 22, 2024. (Total: 46599 publications since 2009)
Records 1-30 (of 43 Records) |
Query Trace: Luquez C [original query] |
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Unintended consequences: Renaming botulinum neurotoxin-producing species of clostridium and related species
Lúquez C , Halpin JL , Dykes J . Toxicon 2023 224 107036 Botulinum neurotoxin-producing species of Clostridium are highly diverse. Clostridium botulinum could represent at least four different species of Clostridium. In addition, strains that do not produce botulinum neurotoxin are closely related to toxigenic strains, probably representing the same species. Although reclassification of these organisms has been proposed in the past, their species names have remained unchanged, mainly because of the premise that changing names of medically relevant organisms might cause confusion in the healthcare and scientific community. In this review, we discuss the possible unintended consequences of reclassifying botulinum neurotoxin-producing species of Clostridium, which are of public health, medical, and biodefense interest. |
Draft Genome Sequences of 20 Clostridium botulinum Type A Isolates from Foodborne Botulism Outbreaks.
Halpin JL , Gómez GA , Dykes JK , Lúquez C . Microbiol Resour Announc 2023 12 (1) e0086822 Here, we present 20 draft genome sequences of Clostridium botulinum type A isolates originating from foodborne outbreaks in the United States and Ethiopia. Publicly available genomes enhance our understanding of C. botulinum genomics and are an asset in bioterrorism preparedness. |
Clostridium botulinum Type B Isolated From a Wound Botulism Case Due to Injection Drug Use Resembles Other Local Strains Originating From Hawaii.
Halpin JL , Foltz V , Dykes JK , Chatham-Stephens K , Lúquez C . Front Microbiol 2021 12 678473 Clostridium botulinum produces botulinum neurotoxin (BoNT), which can lead to death if untreated. In the United States, over 90% of wound botulism cases are associated with injection drug use of black tar heroin. We sought to determine the phylogenetic relatedness of C. botulinum isolated from an injection drug use wound botulism case and isolates from endogenous infant botulism cases in Hawaii. Nineteen C. botulinum type B isolates from Hawaii and one type B isolate from California were analyzed by whole-genome sequencing. The botulinum toxin gene (bont) subtype was determined using CLC Genomics Workbench, and the seven-gene multi-locus sequence type (MLST) was identified by querying PubMLST. Mashtree and pairwise average nucleotide identity were used to find nearest neighbors, and Lyve-SET approximated a phylogeny. Eighteen of the isolates harbored the bont/B5 gene: of those, 17 were classified as sequence type ST36 and one was classified as ST104. A single isolate from Hawaii harbored bont/B1 and was determined to belong to ST110, and the isolate from California harbored bont/B1 and belonged to ST30. A tree constructed with Lyve-SET showed a high degree of homology among all the Hawaiian C. botulinum isolates that harbor the bont/B5 gene. Our results indicate that the bont/B-expressing isolates recovered from Hawaii are closely related to each other, suggesting local contamination of the drug paraphernalia or the wound itself with spores rather than contamination of the drug at manufacture or during transport. These findings may assist in identifying interventions to decrease wound botulism among persons who inject drugs. |
Genomic Characterization of Strains From a Cluster of Infant Botulism Type A in a Small Town in Colorado, United States.
Gladney L , Halpin JL , Lúquez C . Front Microbiol 2021 12 688240 Three cases of infant botulism were reported in a small Colorado town between 1981 and 1984. The first two cases occurred in 1981, 6 months apart, and the third case occurred in 1984. Clostridium botulinum type A was isolated from stool of all three case patients and from environmental samples of the patient's homes. An epidemiological investigation and follow-up study were conducted from 1981 to 1986 and concluded the cases were likely related. In this study, we sought to determine whether the C. botulinum type A clinical isolates were related to each other and to isolates obtained from environmental samples. We performed whole genome sequencing (WGS) for 17 isolates associated with this potential cluster of infant botulism. Fifteen isolates were confirmed to be C. botulinum type A(B) and contained botulinum toxin gene subtypes A1 and B5 by WGS; these strains formed a monophyletic cluster in a phylogeny and were considered closely related to each other (0-18 high-quality single-nucleotide polymorphisms), but distinct from other C. botulinum type A(B) in Colorado and elsewhere in the United States. Results of our study suggest that the three infant botulism cases could have represented a cluster due to a C. botulinum type A(B) strain present in the environment. |
Foodborne botulism outbreaks in the United States, 2001-2017
Lúquez C , Edwards L , Griffin C , Sobel J . Front Microbiol 2021 12 713101 Foodborne botulism is an intoxication caused by ingestion of food containing botulinum neurotoxin. Cases of foodborne botulism are usually sporadic (single, unrelated) but outbreaks of two or more cases occur. In this mini-review we will examine the following for the period 2001-2017, in the United States: botulism surveillance data, outbreaks of botulism affecting 10 or more people, and the public health preparedness and response approach. |
Clinical Guidelines for Diagnosis and Treatment of Botulism, 2021
Rao AK , Sobel J , Chatham-Stephens K , Luquez C . MMWR Recomm Rep 2021 70 (2) 1-30 Botulism is a rare, neurotoxin-mediated, life-threatening disease characterized by flaccid descending paralysis that begins with cranial nerve palsies and might progress to extremity weakness and respiratory failure. Botulinum neurotoxin, which inhibits acetylcholine release at the neuromuscular junction, is produced by the anaerobic, gram-positive bacterium Clostridium botulinum and, rarely, by related species (C. baratii and C. butyricum). Exposure to the neurotoxin occurs through ingestion of toxin (foodborne botulism), bacterial colonization of a wound (wound botulism) or the intestines (infant botulism and adult intestinal colonization botulism), and high-concentration cosmetic or therapeutic injections of toxin (iatrogenic botulism). In addition, concerns have been raised about the possibility of a bioterrorism event involving toxin exposure through intentional contamination of food or drink or through aerosolization. Neurologic symptoms are similar regardless of exposure route. Treatment involves supportive care, intubation and mechanical ventilation when necessary, and administration of botulinum antitoxin. Certain neurological diseases (e.g., myasthenia gravis and Guillain-Barré syndrome) have signs and symptoms that overlap with botulism. Before the publication of these guidelines, no comprehensive clinical care guidelines existed for treating botulism. These evidence-based guidelines provide health care providers with recommended best practices for diagnosing, monitoring, and treating single cases or outbreaks of foodborne, wound, and inhalational botulism and were developed after a multiyear process involving several systematic reviews and expert input. |
Notes from the field: Botulism type E after consumption of salt-cured fish - New Jersey, 2018
Ganapathiraju PV , Gharpure R , Thomas D , Millet N , Gurrieri D , Chatham-Stephens K , Dykes J , Luquez C , Dinavahi P , Ganapathiraju S , Roger S , Abbasi D , Higgins N , Loftus F , Trivedi M . MMWR Morb Mortal Wkly Rep 2019 68 (44) 1008-1009 On October 25, 2018, at 2:15 a.m., a woman aged 30 years and her mother, aged 55 years, both of Egyptian descent, arrived at an emergency department in New Jersey in hypotensive shock after 16 hours of abdominal pain, vomiting, and diarrhea. The daughter also reported blurry vision and double vision (diplopia), shortness of breath, chest pain, and difficulty speaking. She appeared lethargic and had ophthalmoplegia and bilateral ptosis. Both women were admitted to the hospital. The mother improved after fluid resuscitation, but the daughter required vasopressor support in the intensive care unit. Although the mother did not have evidence of cranial nerve involvement on admission, during the next 24 hours, she developed dysphagia and autonomic dysfunction with syncope and orthostasis and was transferred to the intensive care unit as her symptoms progressively worsened similar to those of her daughter. |
Molecular Characterization of Clostridium botulinum Harboring the bont/B7 Gene.
Halpin JL , Dykes JK , Katz L , Centurioni DA , Perry MJ , Egan CT , Luquez C . Foodborne Pathog Dis 2019 16 (6) 428-433 Clostridium botulinum produces botulinum neurotoxin (BoNT), which is the causative agent of botulism, a rare but serious disease that can result in death if not treated. Infant botulism occurs when C. botulinum colonizes the intestinal tract of infants and produces BoNT. It has been proposed that infants under the age of 1 year are uniquely susceptible to colonization by C. botulinum as their intestinal microbiota is not fully developed and provides little competition, allowing C. botulinum to thrive and produce BoNT in the gut. There are seven well-characterized serotypes (A-G) of BoNT identified by the ability of specific antitoxins to neutralize BoNTs. Molecular technology has allowed researchers to narrow these further into subtypes based on nucleic acid sequences of the botulinum toxin (bont) gene. One of the most recently recognized subtypes for bont/B is subtype bont/B7. We identified through whole genome sequencing five C. botulinum isolates harboring bont/B7 from CDC's strain collection, including patient isolates and an epidemiologically linked isolate from an opened infant formula container. In this study, we report the results of whole genome sequencing analysis of these C. botulinum subtype bont/B7 isolates. Average nucleotide identity and high quality single nucleotide polymorphism (hqSNP) analysis resulted in two major clades. The epidemiologically linked isolates differed from each other by 2-6 hqSNPs, and this clade separated from the other isolates by 95-119 hqSNPs, corroborating available epidemiological evidence. |
Notes from the Field: Botulism outbreak associated with home-canned peas - New York City, 2018
Bergeron G , Latash J , Da Costa-Carter CA , Egan C , Stavinsky F , Kileci JA , Winstead A , Zhao B , Perry MJ , Chatham-Stephens K , Sarpel D , Hughes S , Conlon MA , Edmunds S , Mohanraj M , Rakeman JL , Centurioni DA , Luquez C , Chiefari AK , Harper S . MMWR Morb Mortal Wkly Rep 2019 68 (10) 251-252 On June 6, 2018, at 1:30 p.m., the New York City Department of Health and Mental Hygiene was notified of three related women who had arrived at a hospital 4 hours earlier for evaluation for acute nausea, dizziness, blurred vision, slurred speech, ptosis, thick-feeling tongue, and shortness of breath. Two patients developed respiratory failure, requiring intubation and mechanical ventilation in the emergency department, and the third patient was intubated at 7 p.m. that evening. The combination of cranial nerve palsies and respiratory failure in multiple patients suggested botulism, a paralytic illness caused by botulinum neurotoxin (BoNT), most commonly produced by Clostridium botulinum. |
Draft Genome Sequence of a Clostridium botulinum Isolate from Thailand Harboring the Subtype bont /B8 Gene.
Halpin JL , Wangroongsarb P , Jittaprasartsin C , Dykes JK , Luquez C . Microbiol Resour Announc 2019 8 (5) In 2010, a Clostridium botulinum type B isolate was recovered from fermented soybeans during a foodborne botulism investigation. Molecular investigation of the botulinum neurotoxin (bont) gene operon determined that the sequence was a new subtype, denoted B8. Here, we describe the draft whole-genome sequence of the organism. |
Draft Genome Sequences for Dual-Toxin-Producing Clostridium botulinum Strains.
Halpin JL , Dykes JK , Luquez C . Microbiol Resour Announc 2019 8 (4) Here, we present draft genome sequences for three Clostridium botulinum strains that produce multiple botulinum toxin serotypes. Strains that produce two toxins are rare; however, one of these strains produces subtype B5 and F2 toxins, and two of the strains produce subtype A4 and B5 toxins. |
A case of localized, unilateral (cephalic) wound botulism
Jacobs Slifka K , Harris JA , Nguyen V , Luquez C , Tiwari T , Rao AK . Clin Infect Dis 2017 66 S95-s98 We describe a rare presentation of botulism originally presenting with exclusively unilateral cranial nerve deficits following a puncture wound to the face. Cephalic tetanus was initially suspected but laboratory testing confirmed botulism. Botulism caused by local diffusion of toxin from a contaminated head wound can be confused with cephalic tetanus. |
Outbreak of botulism due to illicit prison-brewed alcohol: Public health response to a serious and recurrent problem
Rao AK , Walters M , Hall J , Guymon C , Garden R , Sturdy P , Thurston D , Smith L , Dimond M , Vitek D , Bogdanow L , Hill M , Lin NH , Luquez C , Griffin PM . Clin Infect Dis 2017 66 S85-s91 Background: Botulism is a rare, sometimes lethal neuroparalytic illness. On 2 October 2011, an inmate at prison A developed symptoms compatible with botulism after drinking pruno, an illicit, prison-brewed alcoholic beverage. Additional illnesses were identified within several days. We conducted an investigation to determine the cause and extent of the outbreak. Methods: A case was defined as signs or symptoms of botulism in a prison A inmate with onset during 30 September-9 October 2011. Cases were identified through medical evaluations and interviews with inmates about recent pruno consumption. Laboratory testing was performed for Clostridium botulinum and botulinum neurotoxin. Ingredients, preparation, and sharing of the implicated pruno were investigated. Results: Eight prisoners developed botulism; all drank pruno made with a potato. Three received mechanical ventilation. Culture of fluid from a sock that inmates reported using to filter the implicated pruno yielded C. botulinum type A. The implicated batch may have been shared between cells during delivery of meal trays. Challenges of the investigation included identifying affected inmates, overcoming inaccuracies in histories, and determining how the illicit beverage was shared. Costs to taxpayers were nearly $500000 in hospital costs alone. Conclusions: Pruno made with potato has emerged as an important cause of botulism in the United States. This public health response illustrates the difficulties of investigating botulism in correctional facilities and lessons learned for future investigations. |
Type F infant botulism: Investigation of recent clusters and overview of this exceedingly rare disease
Halpin AL , Khouri JM , Payne JR , Nakao JH , Cronquist A , Kalas N , Mohr M , Osborne M , O'Dell S , Luquez C , Klontz KC , Sobel J , Rao AK . Clin Infect Dis 2017 66 S92-s94 From 1976 to 2016, neurotoxigenic Clostridium baratii type F caused 18 (<0.5%) reported US infant botulism cases. Six cases occurred during 2012-2013; no common source was identified. Type F infant botulism mostly occurs in very young infants and typically presents more rapidly and severely than illness caused by types A and B botulinum neurotoxin. |
Finished Whole-Genome Sequences of Clostridium butyricum Toxin Subtype E4 and Clostridium baratii Toxin Subtype F7 Strains.
Halpin JL , Hill K , Johnson SL , Bruce DC , Shirey TB , Dykes JK , Luquez C . Genome Announc 2017 5 (29) Clostridium butyricum and Clostridium baratii species have been known to produce botulinum toxin types E and F, respectively, which can cause botulism, a rare but serious neuroparalytic disease. Here, we present finished genome sequences for two of these clinically relevant strains. |
Pulsotype Diversity of Clostridium botulinum Strains Containing Serotypes A and/or B Genes.
Halpin JL , Joseph L , Dykes JK , McCroskey L , Smith E , Toney D , Stroika S , Hise K , Maslanka S , Luquez C . Foodborne Pathog Dis 2017 14 (9) 494-501 Clostridium botulinum strains are prevalent in the environment and produce a potent neurotoxin that causes botulism, a rare but serious paralytic disease. In 2010, a national PulseNet database was established to curate C. botulinum pulsotypes and facilitate epidemiological investigations, particularly for serotypes A and B strains frequently associated with botulism cases in the United States. Between 2010 and 2014 we performed pulsed-field gel electrophoresis (PFGE) using a PulseNet protocol, uploaded the resulting PFGE patterns into a national database, and analyzed data according to PulseNet criteria (UPGMA clustering, Dice coefficient, 1.5% position tolerance, and 1.5% optimization). A retrospective data analysis was undertaken on 349 entries comprised of type A and B strains isolated from foodborne and infant cases to determine epidemiological relevance, resolution of the method, and the diversity of the database. Most studies to date on the pulsotype diversity of C. botulinum have encompassed very small sets of isolates; this study, with over 300 isolates, is more comprehensive than any published to date. Epidemiologically linked isolates had indistinguishable patterns, except in four instances and there were no obvious geographic trends noted. Simpson's Index of Diversity (D) has historically been used to demonstrate species diversity and abundance within a group, and is considered a standard descriptor for PFGE databases. Simpson's Index was calculated for each restriction endonuclease (SmaI, XhoI), the pattern combination SmaI-XhoI, as well as for each toxin serotype. The D values indicate that both enzymes provided better resolution for serotype B isolates than serotype A. XhoI as the secondary enzyme provided little additional discrimination for C. botulinum. SmaI patterns can be used to exclude unrelated isolates during a foodborne outbreak, but pulsotypes should always be considered concurrently with available epidemiological data. |
Finished Whole-Genome Sequences of Two Clostridium botulinum Type A(B) Isolates.
Halpin JL , Hill K , Johnson SL , Bruce DC , Shirey TB , Dykes JK , Luquez C . Genome Announc 2017 5 (21) Clostridium botulinum secretes a potent neurotoxin that causes devastating effects when ingested, including paralysis and death if not treated. In the United States, some clinically significant strains produce toxin type A while also harboring a silent B gene. These are the first two closed genome sequences published for this subset. |
Finished Whole-Genome Sequence of Clostridium argentinense Producing Botulinum Neurotoxin Type G.
Halpin JL , Hill K , Johnson SL , Bruce DC , Shirey TB , Dykes JK , Luquez C . Genome Announc 2017 5 (21) Here, we present a closed genome sequence for Clostridium argentinense strain 89G, the first strain identified to produce botulinum neurotoxin type G (BoNT/G). Although discovered in 1970, to date, there have been no reference quality sequences publicly available for this species. |
Historical perspectives and guidelines for botulinum neurotoxin subtype nomenclature
Peck MW , Smith TJ , Anniballi F , Austin JW , Bano L , Bradshaw M , Cuervo P , Cheng LW , Derman Y , Dorner BG , Fisher A , Hill KK , Kalb SR , Korkeala H , Lindstrom M , Lista F , Luquez C , Mazuet C , Pirazzini M , Popoff MR , Rossetto O , Rummel A , Sesardic D , Singh BR , Stringer SC . Toxins (Basel) 2017 9 (1) Botulinum neurotoxins are diverse proteins. They are currently represented by at least seven serotypes and more than 40 subtypes. New clostridial strains that produce novel neurotoxin variants are being identified with increasing frequency, which presents challenges when organizing the nomenclature surrounding these neurotoxins. Worldwide, researchers are faced with the possibility that toxins having identical sequences may be given different designations or novel toxins having unique sequences may be given the same designations on publication. In order to minimize these problems, an ad hoc committee consisting of over 20 researchers in the field of botulinum neurotoxin research was convened to discuss the clarification of the issues involved in botulinum neurotoxin nomenclature. This publication presents a historical overview of the issues and provides guidelines for botulinum neurotoxin subtype nomenclature in the future. |
Notes from the field: Botulism outbreak from drinking prison-made illicit alcohol in a federal correctional facility - Mississippi, June 2016
McCrickard L , Marlow M , Self JL , Watkins LF , Chatham-Stephens K , Anderson J , Hand S , Taylor K , Hanson J , Patrick K , Luquez C , Dykes J , Kalb SR , Hoyt K , Barr JR , Crawford T , Chambers A , Douthit B , Cox R , Craig M , Spurzem J , Doherty J , Allswede M , Byers P , Dobbs T . MMWR Morb Mortal Wkly Rep 2017 65 (52) 1491-1492 On June 9, 2016, the Mississippi Poison Control Center and the Mississippi State Department of Health (MSDH) notified CDC of five suspected cases of botulism, a potentially fatal neuroparalytic illness (1), in inmates at a medium-security federal correctional institution (prison A). By June 10, a total of 13 inmates were hospitalized, including 12 in Mississippi and one in Oklahoma (the inmate in Oklahoma had been transferred there after his exposure for reasons unrelated to his illness). MSDH, Oklahoma State Department of Health, Bureau of Prisons, and CDC conducted an investigation to identify the source and scope of the outbreak, and to develop recommendations. | Prison A staff members suspected that an alcoholic beverage, illicitly made by inmates and known as “hooch” or “pruno,” was the source of the outbreak. Among 33 inmates who reported consuming hooch during June 1–19, 2016, a total of 31 (94%) had signs or symptoms suggesting botulism. The median interval from first exposure to symptom onset was 3 days (range = 0–11 days) (Figure). Cases were categorized using modified Council of State and Territorial Epidemiologists definitions. A confirmed case was defined as an illness in an inmate consistent with botulism that began on or after June 1, with botulinum toxin type A detected in a serum or stool specimen or Clostridium botulinum cultured from a stool specimen; a probable case was defined as an illness in an inmate with signs or symptoms of any cranial nerve palsy and extremity weakness that began on or after June 1; and a suspected case was an illness in an inmate with signs or symptoms of any cranial nerve palsy without extremity weakness that began on or after June 1. |
Notes from the field: Clostridium perfringens gastroenteritis outbreak associated with a catered lunch - North Carolina, November 2015
Rinsky JL , Berl E , Greene V , Morrow J , Didomenico A , MacFarquhar J , Gomez GA , Luquez C , Williams C . MMWR Morb Mortal Wkly Rep 2016 65 (46) 1300-1301 During November 2015, the North Carolina Division of Public Health was notified by the Pitt County Health Department (PCHD) that approximately 40 persons who attended a catered company Thanksgiving lunch the previous day were ill with diarrhea and abdominal pain. The North Carolina Division of Public Health and PCHD worked together to investigate the source of illness and implement control measures. Within hours of notification, investigators developed and distributed an online survey to all lunch attendees regarding symptoms and foods consumed and initiated a cohort study. A case of illness was defined as abdominal pain or diarrhea in a lunch attendee with illness onset <24 hours after the event. Risk ratios (RRs) and 95% confidence intervals (CIs) were estimated for all menu items. Among 80 attendees, 58 (73%) completed the survey, including 44 respondents (76%) who reported illnesses meeting the case definition; among these, 41 (93%) reported diarrhea, and 40 (91%) reported abdominal pain. There were no hospitalizations. Symptom onset began a median of 13 hours after lunch (range = 1-22 hours). Risk for illness among persons who ate turkey or stuffing (38 of 44; 86%), which were plated and served together, was significantly higher than risk for illness among those who did not eat turkey or stuffing (six of 14; 43%) (RR = 2.02; 95% CI = 1.09-3.73). |
Characterizing the fecal microbiota of infants with botulism.
Shirey TB , Dykes JK , Luquez C , Maslanka SE , Raphael BH . Microbiome 2015 3 (1) 54 BACKGROUND: Infant botulism is the most prevalent form of botulism in the USA, representing 68.5 % of cases reported from 2001-2012. Infant botulism results when botulinum toxin-producing clostridia (BTPC) colonize the infant gut with concomitant in vivo production of the highly potent botulinum neurotoxin (BoNT). The gut microbiota of infants with botulism is largely uncharacterized; therefore, it remains unclear whether the microbiota profile of these patients are distinct in composition, abundance, or diversity. To address this uncertainty, we employed 16S rRNA gene profiling to characterize the fecal microbiota in 14 stool samples among laboratory-confirmed and non-confirmed infant botulism cases. RESULTS: Seven bacterial phyla were identified among all 14 infant stool samples examined. Compared to samples from non-confirmed cases, the fecal microbiota of infant botulism patients displayed significantly higher Proteobacteria abundance. Of the 20 bacterial families identified, Enterobacteriaceae was significantly more abundant in samples from infants with botulism. Firmicutes abundance and the abundance ratio of Firmicutes/Proteobacteria was significantly lower in samples from infants with botulism. Lactobacillus spp. abundance was notably reduced in 12 of the 14 samples. Clostridium botulinum and Clostridium baratii were identified in low relative abundances in confirmed and non-confirmed samples based on their 16S rRNA gene profiles, although their toxigenicity remained undetermined. No significant differences were observed in the number of operational taxonomic units (OTUs) observed or in fecal microbiota diversity between laboratory-confirmed and non-confirmed samples. Correlations between individual phylum abundances and infant age were variable, and no significant differences were shown in number of OTUs observed or in fecal microbiota diversity between samples delineated by overall mean age. CONCLUSIONS: Significant differences in Proteobacteria, Firmicutes, and Enterobacteriaceae abundances were identified in the fecal microbiota of infants with botulism when compared to samples from non-confirmed cases. Fecal microbiota diversity was not significantly altered in infants with botulism, and a limited presence of BTPC was shown. It could not be determined whether the fecal microbiota profiles shown here were comparable prior to patient illness, or whether they were the direct result of infant botulism. The results of this study do, however, provide a detailed and descriptive observation into the infant gut microbiota after intestinal colonization by BTPC. |
Laboratory Investigation of the First Case of Botulism Caused by Clostridium butyricum Type E Toxin in the United States
Dykes JK , Luquez C , Raphael BH , McCroskey L , Maslanka SE . J Clin Microbiol 2015 53 (10) 3363-5 We report here the laboratory investigation of the first known case of botulism in the United States caused by C. butyricum type E. This investigation demonstrates the importance of extensive microbiological examination of specimens which resulted in the isolation of this organism. |
Notes from the field: large outbreak of botulism associated with a church potluck meal - Ohio, 2015
McCarty CL , Angelo K , Beer KD , Cibulskas-White K , Quinn K , Fijter S , Bokanyi R , Germain ES , Baransi K , Barlow K , Shafer G , Hanna L , Spindler K , Walz E , DiOrio M , Jackson BR , Luquez C , Mahon BE , Basler C , Curran K , Matanock A , Walsh K , Slifka KJ , Rao AK . MMWR Morb Mortal Wkly Rep 2015 64 (29) 802-803 On April 21, 2015, the Fairfield Medical Center (FMC) and Fairfield Department of Health contacted the Ohio Department of Health (ODH) about a patient suspected of having botulism in Fairfield County, Ohio. Botulism is a severe, potentially fatal neuroparalytic illness.* A single case is a public health emergency, because it can signal an outbreak. Within 2 hours of health department notification, four more patients with similar clinical features arrived at FMC's emergency department. Later that afternoon, one patient died of respiratory failure shortly after arriving at the emergency department. All affected persons had eaten at the same widely attended church potluck meal on April 19. CDC's Strategic National Stockpile sent 50 doses of botulinum antitoxin to Ohio. FMC, the Fairfield Department of Health, ODH, and CDC rapidly responded to confirm the diagnosis, identify and treat additional patients, and determine the source. |
A Novel Botulinum Toxin, Previously Reported as Serotype H, has a Hybrid Structure of Known Serotypes A and F that is Neutralized with Serotype A Antitoxin.
Maslanka SE , Luquez C , Dykes JK , Tepp WH , Pier CL , Pellett S , Raphael BH , Kalb SR , Barr JR , Rao A , Johnson EA . J Infect Dis 2015 213 (3) 379-85 Botulism is a potentially fatal paralytic disease caused by the action of botulinum neurotoxin (BoNT) on nerve cells. There are 7 known serotypes (A through G) of BoNT and up to 40 genetic variants. Clostridium botulinum strain IBCA10-7060 was recently reported to produce BoNT serotype B (BoNT/B) and a novel BoNT, designated by the authors as BoNT/H. The botulinum neurotoxin gene (bont) sequence of BoNT/H was analyzed against known related bont sequences. Genetic analysis suggested that BoNT/H has a hybrid-like structure containing regions of similarity to BoNT/A1 and /F5. This novel BoNT was serologically characterized by the mouse neutralization assay and a neuronal cell-based assay. The toxic effects of this hybrid-like BoNT were completely eliminated by existing serotype A antitoxin including those contained in multivalent therapeutic antitoxin products that are the mainstay for human botulism treatment. |
Functional characterization of botulinum neurotoxin serotype H as a hybrid of known serotypes F and A (BoNT F/A)
Kalb SR , Baudys J , Raphael BH , Dykes JK , Luquez C , Maslanka SE , Barr JR . Anal Chem 2015 87 (7) 3911-7 A unique strain of Clostridium botulinum (IBCA10-7060) was recently discovered which produces two toxins: botulinum neurotoxin (BoNT) serotype B and a novel BoNT reported as serotype H. Previous molecular assessment showed that the light chain (LC) of the novel BoNT most resembled the bont of the light chain of known subtype F5, while the C-terminus of the heavy chain (HC) most resembled the binding domain of serotype A. We evaluated the functionality of both toxins produced in culture by first incorporating an immunoaffinity step using monoclonal antibodies to purify BoNT from culture supernatants and tested each immune-captured neurotoxin with full-length substrates vesicle-associated membrane protein 2 (VAMP-2), synaptosomal-associated protein 25 (SNAP-25), syntaxin, and shortened peptides representing the substrates. The BoNT/B produced by this strain behaved as a typical BoNT/B, having immunoaffinity for anti-B monoclonal antibodies and cleaving both full length VAMP-2 and a peptide based on the sequence of VAMP-2 in the expected location. As expected, there was no activity toward SNAP-25 or syntaxin. The novel BoNT demonstrated immunoaffinity for anti-A monoclonal antibodies but did not cleave SNAP-25 as expected for BoNT/A. Instead, the novel BoNT cleaved VAMP-2 and VAMP-2-based peptides in the same location as BoNT/F5. This is the first discovery of a single botulinum neurotoxin with BoNT/A antigenicity and BoNT/F light chain function. This work suggests that the newly reported serotype H may actually be a hybrid of previously known BoNT serotype A and serotype F, specifically subtype F5. |
Molecular subtyping of Clostridium botulinum by pulsed-field gel electrophoresis.
Luquez C , Joseph LA , Maslanka SE . Methods Mol Biol 2015 1301 103-13 Pulsed-field gel electrophoresis (PFGE) has been extensively used to estimate the genetic diversity of Clostridium botulinum. In addition, PFGE is the standard method for investigating foodborne outbreaks associated with various enteric pathogens, including C. botulinum. PFGE can be used to exclude a suspected but not confirmed food source when the patterns of the food and clinical isolates are different. Indistinguishable PFGE patterns may also be useful for linking isolates between patients or to a food source, but results must be interpreted within an epidemiological context to ensure isolates are truly related. Here, we describe a standardized laboratory protocol for molecular subtyping of C. botulinum by PFGE. |
Detection of botulinum toxins A, B, E, and F in foods by Endopep-MS
Kalb SR , Krilich JC , Dykes JK , Luquez C , Maslanka SE , Barr JR . J Agric Food Chem 2015 63 (4) 1133-1141 Botulism is caused by exposure to botulinum neurotoxins (BoNTs). BoNTs are proteins secreted by some species of clostridia; these neurotoxins are known to interfere with nerve impulse transmission, thus causing paralysis. Botulism may be contracted through consumption of food either naturally or intentionally contaminated with BoNT. The human lethal dose of BoNT is not known but is estimated to be between 0.1 and 70 mug; thus, it is important to be able to detect small amounts of this toxin in foods to ensure food safety and to identify the source of an outbreak. Our laboratory previously reported on the development of Endopep-MS, a mass-spectrometric-based endopeptidase method for the detection and differentiation of BoNT. This method can detect BoNT at levels below the historic standard mouse bioassay in clinical samples such as serum, stool, and culture supernatants. We have now expanded this assay to detect BoNT in over 50 foods including representative products that were involved in actual botulism investigations. The foods tested by the Endopep-MS included those with various acidities, viscosities, and fat levels. Dairy and culturally diverse products were also included. This work demonstrates that the Endopep-MS method can be used to detect BoNT/A, /B, /E, and /F in foods at levels spiked below that of the limit of detection of the mouse bioassay. Furthermore, we successfully applied this method to investigate several foods associated with botulism outbreaks. |
First report of an infant botulism case due to Clostridium botulinum type Af
de Jong LI , Fernandez RA , Pareja V , Giaroli G , Guidarelli SR , Dykes JK , Luquez C . J Clin Microbiol 2014 53 (2) 740-2 Most infant botulism cases worldwide are due to botulinum toxin types A and B. Rarely, Clostridium botulinum strains that produce two serotypes (Ab, Ba, and Bf) have also been isolated from infant botulism cases. This is the first reported case of infant botulism due to C. botulinum type Af worldwide. |
Distinguishing highly-related outbreak-associated Clostridium botulinum type A(B) strains
Raphael BH , Shirey TB , Luquez C , Maslanka SE . BMC Microbiol 2014 14 192 BACKGROUND: In the United States, most Clostridium botulinum type A strains isolated during laboratory investigations of human botulism demonstrate the presence of an expressed type A botulinum neurotoxin (BoNT/A) gene and an unexpressed BoNT/B gene. These strains are designated type A(B). The most common pulsed-field gel electrophoresis (PFGE) pattern in the C. botulinum PulseNet database is composed of A(B) strains. The purpose of this study was to evaluate the ability of genome sequencing and multi-loci variable number of tandem repeat analysis (MLVA) to differentiate such strains. RESULTS: The genome sequences of type A(B) strains evaluated in this study are closely related and cluster together compared to other available C. botulinum Group I genomes. In silico multilocus sequence typing (MLST) analysis (7-loci) was unable to differentiate any of the type A(B) strains isolated from seven different outbreak investigations evaluated in this study. A 15-locus MLVA scheme demonstrated an improved ability to differentiate these strains, however, repeat unit variation among the strains was restricted to only two loci. Reference-free single nucleotide polymorphism (SNP) analysis demonstrated the ability to differentiate strains from all of the outbreaks examined and a non-outbreak associated strain. CONCLUSIONS: This study confirms that type A(B) strains that share the same PFGE pattern also share closely-related genome sequences. The lack of a complete type A(B) strain representative genome sequence hinders the ability to assemble genomes by reference mapping and analysis of SNPs at pre-identified sites. However, compared to other methods evaluated in this study, a reference-free SNP analysis demonstrated optimal subtyping utility for type A(B) strains using de novo assembled genome sequences. |
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