cereulide and Diarrhea

Topics: Animals; Bacillus cereus; Bacterial Proteins; Depsipeptides; Diarrhea; Enterotoxins; Foodborne Diseases; Gene Expression Regulation, Bacterial; Gram-Positive Bacterial Infections; Hemolysin Proteins; Host-Pathogen Interactions; Humans; Virulence; Vomiting

Bacillus cereus is the causative agent of two distinct forms of gastroenteritic disease connected to food-poisoning. It produces one emesis-causing toxin and three enterotoxins that elicit diarrhea. Due to changing lifestyles and eating habits, B. cereus is responsible for an increasing number of food-borne diseases in the industrial world. In the past, most studies concentrated on the diarrhoeal type of food-borne disease, while less attention has been given to the emetic type of the disease. The toxins involved in the diarrhoeal syndrome are well-known and detection methods are commercially available, whereas diagnostic methods for the emetic type of disease have been limited. Only recently, progress has been made in developing identification methods for emetic B. cereus and its corresponding toxin. We will summarize the data available for the emetic type of the disease and discuss some new insights in emetic strain characteristics, diagnosis, and toxin synthesis.

Topics: Bacillus cereus; Bacterial Toxins; Base Sequence; Depsipeptides; Diarrhea; DNA, Bacterial; Enterotoxins; Foodborne Diseases; Humans; Molecular Sequence Data; Vomiting

Bacillus cereus is a pathogen related with diarrhoeal or emetic food poisoning cases, of which the latter caused by the cereulide-producing isolates are more severe with several reported lethal cases. It is therefore necessary to develop an effective strategy to prevent the propagation of B. cereus in the food supply. In this study, three autolysins from the cereulide-producing B. cereus group isolates, LysIS075, LysF8819.1 and LysCER057, were identified and characterized. The results showed that the three autolysins were highly lytic and bactericidal to the tested cereulide-producing B. cereus group strains and cross-lytic against other tested B. cereus group strains, and they could inhibit the spore germination and propagation of their tested derived emetic strains. Physical and chemical characterization showed that all the three autolysins were alkalophilic with the optimal activity at pH9.0 or 9.5 with one exception of LysF8819.1 also having significant lytic activity at pH5.0, and they all had relative strong lytic activity at 37-50°C during the 30minute assay. However, LysCER057 showed relative susceptibility to thermo-condition. Remarkably, the separate or cock-tail addition of the three autolysins in food matrices (milk and rice porridge) showed effective bactericidal activity within the tested 2h. All the results revealed that the three autolysins might be potential candidates to control emetic B. cereus strains in different applications.

Topics: Bacillus cereus; Bacterial Proteins; Bacterial Toxins; Depsipeptides; Diarrhea; Food Contamination; Food Microbiology; Humans; N-Acetylmuramoyl-L-alanine Amidase

Three Bacillus cereus strains isolated from an outbreak of food poisoning caused by the consumption of fermented black beans (douchi) containing B. cereus is described. The outbreak involved 139 persons who had nausea, vomiting, and diarrhea. The strains were isolated from vomit and the unprepared douchi. Two of the strains produced the emetic toxin cereulide, as evidenced by polymerase chain reaction analysis for the presence of the nonribosomal synthetase cluster responsible for the synthesis of cereulide and by chemical analysis by high-performance liquid chromatography-mass spectrometry. These two strains belong to genetic group III of B. cereus, and multiple locus sequence typing revealed that the type was ST26, as a major part of B. cereus emetic strains. One of these strains produced significantly more cereulide at 37°C than the type cereulide producer (F4810/72), and it was also able to produce the toxin at 40°C and 42°C. The third strain belongs to genetic group IV, and it is a new multiple locus sequence type closely related to strains that are cytotoxic and enterotoxigenic. It possesses genes for hemolysin BL, nonhemolytic enterotoxin, and cytotoxin K2; however, it varies from the majority of strains possessing genes for hemolysin BL by not being hemolytic. Thus, two B. cereus strains producing the emetic toxin cereulide and a strain producing enterotoxins might have been involved in this food-poisoning incident caused by the consumption of a natural fermented food. The ability of one of the strains to produce cereulide at ≥37°C makes it possible that it is produced in the human gut in addition to occurring in the food.

Topics: Bacillus cereus; China; Depsipeptides; Diarrhea; Disease Outbreaks; Enterotoxins; Fabaceae; Fermentation; Food Contamination; Food Microbiology; Foodborne Diseases; Humans

The immunochromatographic assay, which targets a marker protein co-expressed during the synthesis of cereulide by an emetic Bacillus cereus strain, was used for easily, rapidly and specifically identifying the emetic strains among B. cereus strains from various materials associated with food poisonings. All 50 of the emetic strains showed a positive reaction to the assay, but all 50 diarrheal strains had a negative reaction. The bacterial counts of 108 cfu/ml in enrichment broth and 109 cfu/ml in food-containing enrichment were required for the identification of emetic B. cereus. The present assay could identify easily and specifically the emetic type of B. cereus within 30 min by a pure culture without special techniques and instruments.

Topics: Bacillus cereus; Bacterial Load; Biological Assay; Biomarkers; Depsipeptides; Diarrhea; Emetics; Food Microbiology; Foodborne Diseases

Bacillus cereus can cause diarrheal and emetic types of food poisoning but little study has been done on emetic type of food poisoning in Korea. The objective of this study was to report on the emetic type of food poisoning associated with B. cereus in Korea. The toxin gene profile, toxin production, and antibiotic resistance of B. cereus isolates were investigated in this study. B. cereus was detected in three out of four samples, while the other food poisoning bacteria were not detected. All isolates (KUGH 10, 11, and 12) presented nhe A, B, and C diarrheal toxin genes (755, 743, and 683 bp), detected using NHA, NHB, and NHC primers, and ces emetic toxin gene (1271 bp), detected using CES primer, and produced nonhemolytic enterotoxin and emetic toxin (cereulide), detected using immunochemical assay and high performance liquid chromotography/mass spectrometry (HPLC/MS) analysis. All emetic-associated isolates were resistant to beta-lactam antibiotics. Most important finding in this study was that the risk of emetic-type B. cereus food poisoning has existed in Korea. This suggested that the food poisoning caused by B. cereus producing emetic and diarrheal toxins should be constantly evaluated to prevent misdiagnosis between emetic and diarrheal types of food poisoning.

Topics: Anti-Bacterial Agents; Bacillaceae Infections; Bacillus cereus; beta-Lactams; Chromatography, High Pressure Liquid; Depsipeptides; Diagnosis, Differential; Diarrhea; Disease Outbreaks; Drug Resistance, Bacterial; Enterotoxins; Foodborne Diseases; Humans; Korea; Microbial Sensitivity Tests; Oryza; Protein Isoforms; Seeds; Spectrometry, Mass, Electrospray Ionization; Vomiting

Cereulide is a K(+) ionophore cytotoxic and mitochondriotoxic to primary cells and cell lines of human and other mammalian origins. It is a heat-stable, highly lipophilic (logK(ow) 5.96) peptide (1152 g mol(-1)) produced by certain strains of Bacillus cereus, a bacterium connected to emetic food poisonings. In this study the pancreatic toxicity of purified cereulide, and cereulide-containing bacterial extracts, was studied using fetal porcine Langerhans islets in culture. Exposure to 1ngml(-1) of purified cereulide caused necrotic cell death of the islet cells impairing their insulin content within 2 days. Cell extracts of cereulide-positive B. cereus strains connected to food poisoning or isolated from foodstuffs were toxic, corresponding to their measured cereulide content. Extracts of B. cereus strains producing or not producing the B. cereus diarrheal toxin, but no cereulide, were tolerated by the porcine islet cultures up to concentrations 1000-fold higher compared to extracts from strains containing cereulide, and up to exposure times of 7d. Cereulide thus was identified as the B. cereus-produced substance toxic towards porcine fetal Langerhans islets and beta cells.

Topics: Animals; Bacillus cereus; Bacterial Toxins; Cell Death; Cell Survival; Depsipeptides; Diarrhea; Dose-Response Relationship, Drug; Female; Foodborne Diseases; Insulin; Islets of Langerhans; Pancreas; Swine; Time Factors; Toxicity Tests

An in-depth polyphasic approach was applied to study the population structure of the human pathogen Bacillus cereus. To assess the intraspecific biodiversity of this species, which is the causative agent of gastrointestinal diseases, a total of 90 isolates from diverse geographical origin were studied by genetic [M13-PCR, random amplification of polymorphic DNA (RAPD), multilocus sequence typing (MLST)] and phenetic [Fourier transform Infrared (FTIR), protein profiling, biochemical assays] methods. The strain set included clinical strains, isolates from food remnants connected to outbreaks, as well as isolates from diverse food environments with a well documented strain history. The phenotypic and genotypic analysis of the compiled panel of strains illustrated a considerable diversity among B. cereus connected to diarrhoeal syndrome and other non-emetic food strains, but a very low diversity among emetic isolates. Using all typing methods, cluster analysis revealed a single, distinct cluster of emetic B. cereus strains. The isolates belonging to this cluster were neither able to degrade starch nor could they ferment salicin; they did not possess the genes encoding haemolysin BL (Hbl) and showed only weak or no haemolysis. In contrast, haemolytic-enterotoxin-producing B. cereus strains showed a high degree of heterogeneity and were scattered over different clusters when different typing methods were applied. These data provide evidence for a clonal population structure of cereulide-producing emetic B. cereus and indicate that emetic strains represent a highly clonal complex within a potentially panmictic or weakly clonal background population structure of the species. It may have originated only recently through acquisition of specific virulence factors such as the cereulide synthetase gene.

Topics: Bacillus cereus; Bacterial Proteins; Bacterial Toxins; Bacterial Typing Techniques; Depsipeptides; Diarrhea; Emetics; Evolution, Molecular; Food Microbiology; Foodborne Diseases; Gram-Positive Bacterial Infections; Humans; Molecular Sequence Data; Polymerase Chain Reaction; Random Amplified Polymorphic DNA Technique; Sequence Analysis, DNA; Spectroscopy, Fourier Transform Infrared