Notes from the Field: Fatal Anthrax Pneumonia in Welders and Other Metalworkers Caused by Bacillus cereus Group Bacteria Containing Anthrax Toxin Genes — U.S. Gulf Coast States, 1994–2020

ABSTRACT The Bacillus cereus group comprises nine species, several of which are pathogenic. Differentiating between isolates that may cause disease and those that do not is a matter of public health and economic importance, but it can be particularly challenging due to the high genomic similarity within the group. To this end, we have developed BTyper, a computational tool that employs a combination of (i) virulence gene-based typing, (ii) multilocus sequence typing (MLST), (iii) panC clade typing, and (iv) rpoB allelic typing to rapidly classify B. cereus group isolates using nucleotide sequencing data. BTyper was applied to a set of 662 B. cereus group genome assemblies to (i) identify anthrax-associated genes in non-B. anthracis members of the B. cereus group, and (ii) identify assemblies from B. cereus group strains with emetic potential. With BTyper, the anthrax toxin genes cya, lef, and pagA were detected in 8 genomes classified by the NCBI as B. cereus that clustered into two distinct groups using k-medoids clustering, while either the B. anthracis poly-γ-d-glutamate capsule biosynthesis genes capABCDE or the hyaluronic acid capsule hasA gene was detected in an additional 16 assemblies classified as either B. cereus or Bacillus thuringiensis isolated from clinical, environmental, and food sources. The emetic toxin genes cesABCD were detected in 24 assemblies belonging to panC clades III and VI that had been isolated from food, clinical, and environmental settings. The command line version of BTyper is available at https://github.com/lmc297/BTyper . In addition, BMiner, a companion application for analyzing multiple BTyper output files in aggregate, can be found at https://github.com/lmc297/BMiner . IMPORTANCE Bacillus cereus is a foodborne pathogen that is estimated to cause tens of thousands of illnesses each year in the United States alone. Even with molecular methods, it can be difficult to distinguish nonpathogenic B. cereus group isolates from their pathogenic counterparts, including the human pathogen Bacillus anthracis, which is responsible for anthrax, as well as the insect pathogen B. thuringiensis. By using the variety of typing schemes employed by BTyper, users can rapidly classify, characterize, and assess the virulence potential of any isolate using its nucleotide sequencing data.

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Toxin production potential and the detection of toxin genes among strains of the Bacillus cereus group isolated along the dairy production chain

International Dairy Journal ◽

10.1016/j.idairyj.2007.03.004 ◽

2007 ◽

Vol 17 (10) ◽

pp. 1201-1208 ◽

Cited By ~ 30

Author(s):

Birgitta Svensson ◽

Amanda Monthán ◽

Marie-Hélène Guinebretière ◽

Christophe Nguyen-Thé ◽

Anders Christiansson

Keyword(s):

Bacillus Cereus ◽

Toxin Production ◽

Dairy Production ◽

Production Potential ◽

Bacillus Cereus Group ◽

Production Chain ◽

Toxin Genes

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Multiplex Polymerase Chain Reaction for Detection of Toxin Genes of Bacillus cereus Group Isolated from Meat and Chicken Products

Zagazig Veterinary Journal ◽

10.21608/zvjz.2020.49230.1122 ◽

2020 ◽

Vol 48 (4) ◽

pp. 457-470

Author(s):

Ahlam Gharib ◽

Norhan Abd El-Aziz ◽

Mai Allam

Keyword(s):

Polymerase Chain Reaction ◽

Bacillus Cereus ◽

Multiplex Polymerase Chain Reaction ◽

Chain Reaction ◽

Bacillus Cereus Group ◽

Toxin Genes ◽

Polymerase Chain

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Characterization of two newly isolated bacteriophages PW2 and PW4 and derived endolysins with lysis activity against Bacillus cereus group strains

Virus Research ◽

10.1016/j.virusres.2021.198489 ◽

2021 ◽

pp. 198489

Author(s):

Xiaofu Wan ◽

Peiling Geng ◽

Jiahui Sun ◽

Zhiming Yuan ◽

Xiaomin Hu

Keyword(s):

Bacillus Cereus ◽

Bacillus Cereus Group ◽

Lysis Activity

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Phenotypic properties and genotyping analysis of Bacillus cereus group isolates from dairy and potato products

LWT ◽

10.1016/j.lwt.2021.110853 ◽

2021 ◽

Vol 140 ◽

pp. 110853

Author(s):

Yiying Huang ◽

Steve H. Flint ◽

Shubo Yu ◽

Yu Ding ◽

Jon S. Palmer

Keyword(s):

Bacillus Cereus ◽

Bacillus Cereus Group ◽

Phenotypic Properties ◽

Potato Products

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Development of Rapid Real-Time PCR and Most-Probable-Number Real-Time PCR Assays To Quantify Enterotoxigenic Strains of the Species in the Bacillus cereus Group

Journal of Food Protection ◽

10.4315/0362-028x-70.12.2774 ◽

2007 ◽

Vol 70 (12) ◽

pp. 2774-2781 ◽

Cited By ~ 17

Author(s):

I-CHEN YANG ◽

DANIEL YANG-CHIH SHIH ◽

JAN-YI WANG ◽

TZU-MING PAN

Keyword(s):

Bacillus Cereus ◽

Real Time ◽

Real Time Pcr ◽

Food Samples ◽

Most Probable Number ◽

Pcr Assay ◽

Bacillus Cereus Group ◽

Probable Number ◽

Cooked Rice ◽

Group Detection

Members of the Bacillus cereus group may produce diarrheal enterotoxins and could be potential hazards if they enter the food chain. Therefore, a method capable of detecting all the species in the B. cereus group rather than B. cereus alone is important. We selected nhe as the target and developed a real-time PCR assay to quantify enterotoxigenic strains of the B. cereus group. The real-time PCR assay was evaluated with 60 B. cereus group strains and 28 others. The assay was also used to construct calibration curves for different food matrices and feces. The assay has an excellent quantification capacity, as proved by its linearity (R2 > 0.993), wide dynamic quantification range (102 to 107 CFU/g for cooked rice and chicken, 103 to 107 CFU/ml for milk, and 104 to 107 CFU/g for feces), and adequate relative accuracy (85.5 to 101.1%). For the low-level contaminations, a most-probable-number real-time PCR assay was developed that could detect as low as 100 CFU/ml. Both assays were tested with real food samples and shown to be considerably appropriate for B. cereus group detection and quantification.

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Bacillus cereus G9241 Makes Anthrax Toxin and Capsule like Highly Virulent B. anthracis Ames but Behaves like Attenuated Toxigenic Nonencapsulated B. anthracis Sterne in Rabbits and Mice

Infection and Immunity ◽

10.1128/iai.00205-11 ◽

2011 ◽

Vol 79 (8) ◽

pp. 3012-3019 ◽

Cited By ~ 31

Author(s):

Melissa K. Wilson ◽

James M. Vergis ◽

Farhang Alem ◽

John R. Palmer ◽

Andrea M. Keane-Myers ◽

...

Keyword(s):

Bacillus Cereus ◽

Lethal Dose ◽

Anthrax Toxin ◽

Mouse Strains ◽

Content Type ◽

Intranasal Inoculation ◽

Anthrax Toxins ◽

Highly Virulent ◽

Subcutaneous Inoculation ◽

New Zealand Rabbits

ABSTRACTBacillus cereusG9241 was isolated from a welder with a pulmonary anthrax-like illness. The organism contains two megaplasmids, pBCXO1 and pBC218. These plasmids are analogous to theBacillus anthracisAmes plasmids pXO1 and pXO2 that encode anthrax toxins and capsule, respectively. Here we evaluated the virulence ofB. cereusG9241 as well as the contributions of pBCXO1 and pBC218 to virulence.B. cereusG9241 was avirulent in New Zealand rabbits after subcutaneous inoculation and attenuated 100-fold compared to the published 50% lethal dose (LD50) values forB. anthracisAmes after aerosol inoculation. A/J and C57BL/6J mice were comparably susceptible toB. cereusG9241 by both subcutaneous and intranasal routes of infection. However, the LD50s forB. cereusG9241 in both mouse strains were markedly higher than those reported forB. anthracisAmes and more like those of the toxigenic but nonencapsulatedB. anthracisSterne. Furthermore,B. cereusG9241 spores could germinate and disseminate after intranasal inoculation into A/J mice, as indicated by the presence of vegetative cells in the spleen and blood of animals 48 h after infection. Lastly,B. cereusG9241 derivatives cured of one or both megaplasmids were highly attenuated in A/J mice. We conclude that the presence of the toxin- and capsule-encoding plasmids pBCXO1 and pBC218 inB. cereusG9241 alone is insufficient to render the strain as virulent asB. anthracisAmes. However, likeB. anthracis, full virulence ofB. cereusG9241 for mice requires the presence of both plasmids.

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Complete Circular Genome Sequences of Three Bacillus cereus Group Strains Isolated from Positive Blood Cultures from Preterm and Immunocompromised Infants Hospitalized in France

Microbiology Resource Announcements ◽

10.1128/mra.00597-21 ◽

2021 ◽

Vol 10 (41) ◽

Author(s):

Mariem Ben Khedher ◽

Fredrik Nindo ◽

Alicia Chevalier ◽

Stéphane Bonacorsi ◽

Gregory Dubourg ◽

...

Keyword(s):

Intensive Care ◽

Bacillus Cereus ◽

Complete Genome ◽

Blood Cultures ◽

Genome Sequences ◽

Bacillus Cereus Group ◽

Illumina Hiseq ◽

Content Type ◽

Long Reads ◽

Oxford Nanopore

We report here the complete genome sequences of three Bacillus cereus group strains isolated from blood cultures from premature and immunocompromised infants hospitalized in intensive care units in three French hospitals. These complete genome sequences were obtained from a combination of Illumina HiSeq X Ten short reads and Oxford Nanopore MinION long reads.

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The Possible Transmission and Potential Enterotoxicity of Bacillus cereus on Lettuce Farms in Five Chinese Provinces

Frontiers in Microbiology ◽

10.3389/fmicb.2021.746632 ◽

2021 ◽

Vol 12 ◽

Author(s):

Yang Qu ◽

Chao Wei ◽

Xiaohang Dai ◽

Yalong Bai ◽

Xin Zhao ◽

...

Keyword(s):

Bacillus Cereus ◽

Potential Risk ◽

Trace Analysis ◽

Multilocus Sequence Typing ◽

Soil Samples ◽

Foodborne Diseases ◽

Human Pathogen ◽

Toxin Genes ◽

Positive Rate ◽

Sequence Types

Bacillus cereus is a well-characterized human pathogen that produces toxins associated with diarrheal and emetic foodborne diseases. To investigate the possible transmission of B. cereus on lettuce farms in China and determine its enterotoxicity, (I) a total of 524 samples (lettuce: 332, soil: 69, water: 57, manure: 57, pesticide: 9) were collected from 46 lettuce farms in five Chinese provinces, (II) multilocus sequence typing (MLST) was used to classify B. cereus isolates and for trace analysis, and (III) the presence of toxin genes and enterotoxins (Hbl and Nhe) was detected in 68 strains. The results showed that one hundred and sixty-one lettuce samples (48.5%) tested positive for B. cereus at levels ranging from 10 to 5.3 × 104 CFU/g. Among the environmental sample categories surveyed, the highest positive rate was that of the pesticide samples at 55.6%, followed by soil samples at 52.2% and manure samples at 12.3%. Moreover, one hundred isolates of B. cereus yielded 68 different sequence types (STs) and were classified into five phylogenetic clades. Furthermore, Nhe toxin genes (nheA, nheB, nheC) were broadly distributed and identified in all 68 strains (100%), while Hbl toxin genes (hblA, hblC, hblD) were present in 61 strains (89.7%), entFM was detected in 62 strains (91.2%), and cytK was found in 29 strains (42.6%). All strains were negative for ces. As for the enterotoxin, Nhe was observed in all 68 isolates carrying nheB, while Hbl was present in 76.5% (52/68) of the strains harboring hblC. This study is the first report of possible B. cereus transmission and of its potential enterotoxicity on lettuce farms in China. The results showed that soil and pesticides are the main sources of B. cereus on lettuce farms in China, and the possible transmission routes are as follows: soil-lettuce, manure-lettuce, pesticide-lettuce, manure-soil-lettuce, and water-manure-soil-lettuce. Furthermore, the B. cereus isolates, whether from lettuce or the environment, pose a potential risk to health.

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