Pulsed field gel electrophoresis of chromosomal DNA reveals a clonal population structure to<i>Bacillus thuringiensis</i>that relates in general to crystal protein gene content

Rivera, Adelaida María Gaviria; Priest, Fergus G.

doi:10.1016/s0378-1097(03)00347-1

Cited by 30 publications

(26 citation statements)

References 23 publications

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“…Most serovars in the Kurstaki and Tolworthi lineages were represented by cognate STs, reinforcing the clonal structure of B. thuringiensis noted in other studies (13,14,42). B. thuringiensis serovar kurstaki strains are widespread lepidopteran pathogens containing Cry1 and Cry2 toxins and are often used for biocontrol in agriculture.…”

Section: Discussionsupporting

confidence: 78%

“…B. thuringiensis serovar kurstaki strains are widespread lepidopteran pathogens containing Cry1 and Cry2 toxins and are often used for biocontrol in agriculture. B. thuringiensis serovars aizawai, galleriae, kenyae, and kumamotoensis similarly contain Cry1 toxins, although the exact compositions of the crystals in these bacteria have not been determined (14). This is a group that apparently undergoes extensive sharing of plasmids since all are Cry1-containing types and yet discrete clones are apparent.…”

Section: Discussionmentioning

confidence: 99%

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Population Structure and Evolution of the Bacillus cereus Group

Priest¹,

Barker²,

Baillie³

et al. 2004

J Bacteriol

342

395

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Representative strains of the Bacillus cereus group of bacteria, including Bacillus anthracis (11 isolates), B. cereus (38 isolates), Bacillus mycoides (1 isolate), Bacillus thuringiensis (53 isolates from 17 serovars), and Bacillus weihenstephanensis (2 isolates) were assigned to 59 sequence types (STs) derived from the nucleotide sequences of seven alleles, glpF, gmk, ilvD, pta, pur, pycA, and tpi. Comparisons of the maximum likelihood (ML) tree of the concatenated sequences with individual gene trees showed more congruence than expected by chance, indicating a generally clonal structure to the population. The STs followed two major lines of descent. Clade 1 comprised B. anthracis strains, numerous B. cereus strains, and rare B. thuringiensis strains, while clade 2 included the majority of the B. thuringiensis strains together with some B. cereus strains. Other species were allocated to a third, heterogeneous clade. The ML trees and split decomposition analysis were used to assign STs to eight lineages within clades 1 and 2. These lineages were defined by bootstrap analysis and by a preponderance of fixed differences over shared polymorphisms among the STs. Lineages were named with reference to existing designations: Anthracis, Cereus I, Cereus II, Cereus III, Kurstaki, Sotto, Thuringiensis, and Tolworthi. Strains from some B. thuringiensis serovars were wholly or largely assigned to a single ST, for example, serovar aizawai isolates were assigned to ST-15, serovar kenyae isolates were assigned to ST-13, and serovar tolworthi isolates were assigned to ST-23, while other serovars, such as serovar canadensis, were genetically heterogeneous. We suggest a revision of the nomenclature in which the lineage and clone are recognized through name and ST designations in accordance with the clonal structure of the population.

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Section: Discussionsupporting

confidence: 78%

Section: Discussionmentioning

confidence: 99%

Population Structure and Evolution of the Bacillus cereus Group

Priest¹,

Barker²,

Baillie³

et al. 2004

J Bacteriol

342

395

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“…Further characterization of the three isolates was performed using repetitive sequence-based PCR (rep-PCR) and pulsedfield gel electrophoresis (PFGE) (10,11,25) of genomic DNA. All three strains displayed the same rep-PCR and PFGE profiles (data not shown).…”

Section: Case Reportmentioning

confidence: 99%

Sudden Death of a Young Adult Associated with Bacillus cereus Food Poisoning

et al. 2011

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A lethal intoxication case, which occurred in Brussels, Belgium, is described. A 20-year-old man died following the ingestion of pasta contaminated with Bacillus cereus. Emetic strains of B. cereus were isolated, and high levels of cereulide (14.8 g/g) were found in the spaghetti meal. CASE REPORTOn 1 October 2008, a 20-year-old man became sick after eating a meal of leftovers of spaghetti with tomato sauce, which had been prepared 5 days before and left in the kitchen at room temperature. After school, he warmed the spaghetti in the microwave oven. Immediately after eating, he left home for his sports activities, but he returned 30 min later because of headache, abdominal pain, and nausea. At his arrival, he vomited profusely for several hours and at midnight had two episodes of watery diarrhea. He did not receive any medication and drank only water. After midnight, he fell asleep. The next morning at 11:00 AM, his parents were worried because he did not get up. When they went to his room, they found him dead.Legal examination determined the time of death, presumably at 4:00 AM, approximately 10 h after ingestion of the suspected meal. An autopsy could not be performed until 5 days later. Macroscopically, brownish and moderately softened liver and ascites (550 ml of citrine liquid) were found. The heart was macroscopically normal. A total lysis of the pancreas was also found, but it could not be excluded that this finding was due to the autopsy delay.Microscopic findings were as follows: moderate centrolobular liver necrosis without inflammatory signs and discrete biliary stasis, significant vascular congestion of the lungs, probably due to acute cardiac insufficiency, significant necrosis from all layers of colon mucosa and submucosa alternating with betterpreserved zones, and mixed intestinal flora but no evidence of invasive bacterial lesions. Significant lysis of the adrenal glands was also reported. The exact cause of death could not be determined by the autopsy because the interpretation of findings was very difficult due to the autopsy delay.Five fecal swabs and two feces samples were taken postmortem, and samples were tested for the presence of Bacillus cereus by growth on mannitol egg yolk polymyxin (MYP) agar. B. cereus was found in only two of the five fecal swabs, and the strains isolated were named ISP321 and ISP322. No B. cereus was cultured from the feces samples. Pasta and tomato sauce samples, the leftovers of the dinner, were also sent for analysis to the National Reference Laboratory for Food-borne Outbreaks (NRLFBO). For enumeration of B. cereus in food samples, the ISO 7932 method (16) was used. Significant B. cereus counts (9.5 ϫ 10 7 CFU/g) were found in the pasta, while B. cereus was absent in the tomato sauce. The strain isolated from the pasta meal was named ISP303.PCR assays that detect the presence of toxin genes were applied to DNA from the pasta isolate (ISP303) and the two human isolates (ISP321 and ISP322), and the results are presented in Table 1. The presence of genes encoding ...

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“…1). 5 fingerprints and toxin production of B. cereus isolates. As listed in the Isolate a column, the following isolates have been deposited in the BCCM/LMG Bacteria Collection (Ghent University, Ghent, Belgium): 5975a (LMG 22728); 5965c (LMG 22729); 5972a (LMG 22730); 5969a (LMG 22731); 5958c (LMG 22732); 5964a (LMG 22733).…”

Section: Case Reportmentioning

confidence: 99%