Bacillus cereus in Packaging Material: Molecular and Phenotypical Diversity Revealed

The Bacillus cereus group has been isolated from soils, water, plants and numerous food products. These species can produce a variety of toxins including several enterotoxins [non-hemolytic enterotoxin (Nhe), hemolysin BL (Hbl), cytotoxin K, and enterotoxin FM], the emetic toxin cereulide and insecticidal Bt toxins. This is the first study evaluating the presence of B. cereus in packaging material. Among 75 different isolates, four phylogenetic groups were detected (II, III, IV, and VI), of which the groups III and IV were the most abundant with 46.7 and 41.3%, respectively. One isolate was affiliated to psychrotolerant group VI. Growth experiments showed a mesophilic predominance. Based on PCR analysis, nhe genes were detectable in 100% of the isolates, while hbl genes were only found in 50.7%. The cereulide encoding gene was found in four out of 75 isolates, no isolate carried a crystal toxin gene. In total, thirteen different toxin gene profiles were identified. We showed that a variety of B. cereus group strains can be found in packaging material. Here, this variety lies in the presence of four phylogenetic groups, thirteen toxin gene profiles, and different growth temperatures. The results suggest that packaging material does not contain significant amounts of highly virulent strains, and the low number of cereulide producing strains is in accordance with other results.

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Analysis of enterotoxigenic Bacillus cereus strains from dried foods using whole genome sequencing, multi-locus sequence analysis and toxin gene prevalence and distribution using endpoint PCR analysis

International Journal of Food Microbiology ◽

10.1016/j.ijfoodmicro.2018.06.016 ◽

2018 ◽

Vol 284 ◽

pp. 31-39 ◽

Cited By ~ 9

Author(s):

Laurenda Carter ◽

Hannah R. Chase ◽

Charles M. Gieseker ◽

Nicholas R. Hasbrouck ◽

Cynthia B. Stine ◽

...

Keyword(s):

Sequence Analysis ◽

Bacillus Cereus ◽

Whole Genome Sequencing ◽

Genome Sequencing ◽

Pcr Analysis ◽

Toxin Gene ◽

Whole Genome ◽

Dried Foods

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Genetic Diversity, Antimicrobial Resistance, and Toxigenic Profiles of Bacillus cereus Strains Isolated from Sunsik

Journal of Food Protection ◽

10.4315/0362-028x.jfp-11-317 ◽

2012 ◽

Vol 75 (2) ◽

pp. 225-230 ◽

Cited By ~ 21

Author(s):

NARI LEE ◽

JE MIN SUN ◽

KYUNG YOON KWON ◽

HYUN JUNG KIM ◽

MINSEON KOO ◽

...

Keyword(s):

Bacillus Cereus ◽

Fruits And Vegetables ◽

Food Poisoning ◽

Pcr Analysis ◽

Contamination Level ◽

Detection Rates ◽

Its Gene ◽

Hemolysin Bl ◽

Encoding Genes ◽

Element Sequence

Bacillus cereus can cause emetic and diarrheal types of food poisoning, but little study has been done on the toxins and toxin-encoding genes of B. cereus strains isolated from Sunsik, a Korean ready-to-eat food prepared from grains, fruits, and vegetables. In this study, 39 unique B. cereus strains were isolated and identified from Sunsik samples, with an average contamination level of 10 to 200 CFU/g. The detection rates of the hblACD, cytK, and bceT genes among all the strains were 48.7, 66.7, and 87.1%, respectively. All 39 B. cereus strains carried nheABC and entFM genes, and 36 strains also had the ces gene, which encodes an emetic toxin. Nonhemolytic enterotoxin and hemolysin BL enterotoxin were produced by 39 and 26 strains, respectively. The strains were separated into 13 profiles based on the presence or absence of toxins and their genes, as determined by antibody tests and PCR analysis. Profile 1 was the largest group, comprising 30.7% (12 of 39) of the B. cereus strains tested; these strains harbored all toxins and their genes. The B. cereus strains were susceptible to most of the antibiotics tested but were highly resistant to β-lactam antibiotics. The repetitive element sequence polymorphism PCR fingerprints of the B. cereus strains were not influenced by the presence of toxin genes or antibiotic resistance profiles. Our results suggest that B. cereus strains from Sunsik could cause either the diarrheal or emetic types of food poisoning because all strains isolated contained at least one toxin and its gene, although the level of B. cereus contamination in Sunsik was low.

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DETECTION OF HBLD TOXIN GENE BY BACILLUS CEREUS ISOLATED FROM MEAT CURRY FOOD SAMPLES IN MALAYSIAN RESTAURANTS

Environment & Ecosystem science ◽

10.26480/ees.02.2020.68.72 ◽

2020 ◽

Vol 4 (2) ◽

pp. 68-72

Author(s):

Marwan Msarah ◽

Ahmed Alsier ◽

Sahilah, A.M.

Keyword(s):

Bacillus Cereus ◽

Foodborne Pathogen ◽

Food Poisoning ◽

Egg Yolk ◽

Food Samples ◽

Blood Agar ◽

Toxin Gene ◽

Biochemical Tests ◽

Isolation Medium ◽

Hemolysin Bl

Bacillus cereus is a ubiquitous foodborne pathogen, can cause food poisoning, leading to infections, have two major types of food poisoning emetic and diarrheal. Foods rich in protein such as meat are associated with foodborne outbreaks of diarrhea caused by B. cereus. The aim of this study is to isolate and identify B. cereus from ready to eat (RTE) meat curry from restaurants in Malaysia and to detect hblD pathogenic gene of B. cereus isolates. Mannitol egg yolk polymyxin agar was used as a selective isolation medium. Commercially available kits and boiling methods were used for DNA extraction, samples acquired from restaurants were examined for the presence of Hemolysin BL gene by polymerase chain reaction (PCR). Among all isolates, twenty-four of B. cereus isolates detected for HBL enterotoxin production by the discontinuous pattern on HBL sheep blood agar then confirmed by biochemical tests. More than 58.33 % of the isolate showed discontinuous hemolysis pattern on HBl blood agar and 29.16% of the samples were shown positive for hblD gene that can cause diarrhea with the size of 807bp on gel. This study demonstrated that RTE meat curry was a potential source for entero-toxigenic B. cereus and the presence of the hblD toxin genes for the HBL complex in the isolates tested were highly associated. Therefore, these meat curry isolates should be regarded as potential toxin producers.

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Potential Enterotoxicity of Phylogenetically Diverse Bacillus cereus Sensu Lato Soil Isolates from Different Geographical Locations

Applied and Environmental Microbiology ◽

10.1128/aem.03032-19 ◽

2020 ◽

Vol 86 (11) ◽

Author(s):

Justyna Malgorzata Drewnowska ◽

Natalia Stefanska ◽

Magdalena Czerniecka ◽

Grzegorz Zambrowski ◽

Izabela Swiecicka

Keyword(s):

Bacillus Cereus ◽

Bacterial Population ◽

Phylogenetic Groups ◽

Content Type ◽

Hot Climate ◽

Hemolysin Bl ◽

Spore Forming Bacteria ◽

Geographical Locations ◽

Bacillus Cereus Sensu Lato

ABSTRACT Bacillus cereus sensu lato comprises Gram-positive spore-forming bacteria producing toxins associated with foodborne diseases. Three pore-forming enterotoxins, nonhemolytic enterotoxin (Nhe), hemolysin BL (Hbl), and cytotoxin K (CytK), are considered the primary factors in B. cereus sensu lato diarrhea. The aim of this study was to determine the potential risk of enterotoxicity among soil B. cereus sensu lato isolates representing diverse phylogroups and originated from different geographic locations with various climates (Burkina Faso, Kenya, Argentina, Kazakhstan, and Poland). While nheA- and hblA-positive isolates were present among all B. cereus sensu lato populations and distributed across all phylogenetic groups, cytK-2-positive strains predominated in geographic regions with an arid hot climate (Africa) and clustered together on a phylogenetic tree mainly within mesophilic groups III and IV. The highest in vitro cytotoxicity to Caco-2 and HeLa cells was demonstrated by the strains clustered within phylogroups II and IV. Overall, our results suggest that B. cereus sensu lato pathogenicity is a comprehensive process conditioned by many intracellular factors and diverse environmental conditions. IMPORTANCE This research offers a new route for a wider understanding of the dependency between pathogenicity and phylogeny of a natural bacterial population, specifically within Bacillus cereus sensu lato, that is widely distributed around the world and easily transferred into food products. Our study indicates differences in the phylogenetic and geographical distributions of potential enterotoxigenic B. cereus sensu lato strains. Hence, these bacilli possess a risk for human health, and rapid testing methods for their identification are greatly needed. In particular, the detection of the CytK enterotoxin should be a supporting strategy for the identification of pathogenic B. cereus sensu lato.

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Spore Prevalence and Toxigenicity of Bacillus cereus and Bacillus thuringiensis Isolates from U.S. Retail Spices

Journal of Food Protection ◽

10.4315/0362-028x.jfp-14-380 ◽

2015 ◽

Vol 78 (3) ◽

pp. 590-596 ◽

Cited By ~ 15

Author(s):

UPASANA HARIRAM ◽

RONALD LABBÉ

Keyword(s):

Bacillus Cereus ◽

The United States ◽

Foodborne Illness ◽

Toxin Gene ◽

Enterotoxin Gene ◽

Responsible Gene ◽

Wide Range ◽

Chromogenic Agar ◽

Hemolysin Bl ◽

Spore Forming Bacteria

Recent incidents of foodborne illness associated with spices as the vehicle of transmission prompted this examination of U.S. retail spices with regard to Bacillus cereus. This study focused on the levels of aerobic-mesophilic spore-forming bacteria and B cereus spores associated with 247 retail spices purchased from five states in the United States. Samples contained a wide range of aerobic-mesophilic bacterial spore counts (< 200 to 8.3 × 107 CFU/g), with 19.1% of samples at levels above 105 CFU/g. For examples, paprika, allspice, peppercorns, and mixed spices had high levels of aerobic spores (>107 CFU/g). Using a novel chromogenic agar, B. cereus and B. thuringiensis spores were isolated from 77 (31%) and 11 (4%) samples, respectively. Levels of B. cereus were <3 to 1,600 MPN/g. Eighty-eight percent of B. cereus isolates and 91%of B. thuringiensis isolates possessed at least one type of enterotoxin gene: HBL (hemolysin BL) or nonhemolytic enterotoxin (NHE). None of the 88 isolates obtained in this study possessed the emetic toxin gene (ces). Using commercially available immunological toxin detection kits, the toxigenicity of the isolates was confirmed. The NHE enterotoxin was expressed in 98% of B. cereus and 91% of B. thuringiensis isolates that possessed the responsible gene. HBL enterotoxin was detected in 87% of B. cereus and 100% of B. thuringiensis PCR-positive isolates. Fifty-two percent of B. cereus and 54% of B. thuringiensis isolates produced both enterotoxins. Ninety-seven percent of B. cereus isolates grew at 12°C, although only two isolates grew well at 9°C. The ability of these spice isolates to form spores, produce diarrheal toxins, and grow at moderately abusive temperatures makes retail spices an important potential vehicle for foodborne illness caused by B. cereus strains, in particular those that produce diarrheal toxins.

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Psychrotrophic properties, toxigenic characteristics, and PFGE profiles of Bacillus cereus isolated from different foods and spices

Ciência Rural ◽

10.1590/0103-8478cr20210166 ◽

2022 ◽

Vol 52 (4) ◽

Author(s):

Hayriye Yeşim Can ◽

Mehmet Elmalı ◽

Alper Karagöz ◽

Hüseyin Burak Dişli

Keyword(s):

Bacillus Cereus ◽

Food Poisoning ◽

Raw Milk ◽

Gene Profiling ◽

Poultry Meat ◽

Pcr Analysis ◽

Toxin Gene ◽

Raw Poultry ◽

Pfge Profiles ◽

Milk And Dairy Products

ABSTRACT: Bacillus cereus is an aerobic and facultatively anaerobic, spore-forming bacterium, and it is found naturally in soil and poses a risk factor for the contamination of food and foodstuffs including cereals, vegetables, spices, ready-to-eat (RTE) foods, meats, milk, and dairy products. This study determined the prevalence of B. cereus in raw poultry meat, raw cow’s milk, cheese, spices, and RTE foods in Hatay province. The study also analysed the psychrotrophic properties, toxigenic characteristics, and pulsed-field gel electrophoresis (PFGE) profiles of the isolates. The levels of contamination with B. cereus determined for cheese, raw milk, RTE foods, spices, and raw poultry meat were 16.6%, 34.2%, 42.8%, 49%, and 55.5%, respectively. B. cereus was isolated from 84 (42%) of the 200 samples analysed and the 84 isolates were verified by PCR analysis targeting the haemolysin gene specific for B. cereus. Of the total isolates, 64 (76.1%) were psychrotrophic. The toxin gene profiling of B. cereus isolates was determined by amplifying the four genes nhe, hbl, cytK, and ces. The nhe and cytK genes were most frequently detected in the isolates, while the hbl and ces genes were not found. In addition, a high genetic relationship between the isolates was detected at a 92% similarity level by PFGE analysis. In conclusion, the occurrence of both psychrotrophic and toxigenic B. cereus strains in this study indicated a potential risk for food spoilage and food poisoning.

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Population structure and toxin gene profiles of Bacillus cereus sensu lato isolated from flour products

FEMS Microbiology Letters ◽

10.1093/femsle/fnz240 ◽

2019 ◽

Vol 366 (20) ◽

Cited By ~ 4

Author(s):

Patrick Kindle ◽

Danai Etter ◽

Roger Stephan ◽

Sophia Johler

Keyword(s):

Risk Assessment ◽

Population Structure ◽

Bacillus Thuringiensis ◽

Bacillus Cereus ◽

Toxin Gene ◽

Type Iii ◽

Gene Profiles ◽

Enterotoxin Genes ◽

Parasporal Crystals ◽

Bacillus Cereus Sensu Lato

ABSTRACT Data on the occurrence, population structure and toxinogenic potential of Bacillus cereus sensu lato isolated from flour is essential to enable improved risk assessment. We aimed to provide data on the occurrence of B. cereus sensu lato in flour products at retail level. In addition, we screened the isolates for Bacillus thuringiensis and Bacillus cytotoxicus and determined population structure and toxin gene profiles. We screened 89 flour products for presence of B. cereus sensu lato, resulting in 75 positive samples (84%). We were able to show that the population structure of members of the B. cereus group isolated from flour is highly diverse. Isolates were assigned to panC types II (4%), III (21%), IV (39%) and V (36%). Production of parasporal crystals characteristic for Bacillus thuringiensis was detected in seven isolates assigned to panC type III, IV and V. No B. cytotoxicus were detected. Two of the isolates harbored ces encoding cereulide, which causes the emetic syndrome. Various enterotoxin genes were found, with all isolates harboring nhe, 75% of isolates harboring hbl and 51% of the isolates harboring cytK-2. Our findings suggest that toxinogenic B. cereus sensu lato are common in flour products at retail level.

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Detection of virulence genes of APEC (avian pathogenic Escherichia coli) isolated from poultry in Noakhali, Bangladesh

Bioresearch Communications ◽

10.3329/brc.v7i1.54253 ◽

2021 ◽

Vol 7 (1) ◽

pp. 967-972

Author(s):

Farzana Ehetasum Hossain ◽

Saiful Islam ◽

Md Aminul Islam ◽

Shariful Islam ◽

Firoz Ahmed

Keyword(s):

Escherichia Coli ◽

Virulence Genes ◽

Virulence Gene ◽

Economic Loss ◽

Point Of View ◽

Pcr Analysis ◽

Avian Pathogenic Escherichia Coli ◽

Gene Profiles ◽

Pathogenic Escherichia Coli ◽

Wide Range

Avian colibacillosis, caused by avian pathogenic Escherichia coli (APEC), is one of the major infectious diseases of poultry that bring about great economic loss for the Bangladesh poultry industry. The present study aimed to determine the virulence genes of avian pathogenic Escherichia coli (APEC) from cases of colibacillosis in poultry at the Noakhali district of Bangladesh. Currently, virulence-associated gene profiles of APEC isolates were investigated by polymerase chain reaction (PCR). A total of 24 (twenty-four) Escherichia coli isolates were collected and presumptively identified from 8 (eight) colibacillosis cases from 4 commercial broiler poultry farms (2 broilers per farm) in Noakhali, Bangladesh. The pathogenesis of Escherichia coli involves a wide range of different virulence genes. At this point, four virulence genes, iutA, hlyF, iroN, and iss were detected by PCR analysis. It has been observed that iutA, iss, hlyF, and iroN genes were found in 7(29.16%), 20(83.33%), 22(91.66%), and 24(100%) APEC isolates respectively. Furthermore, out of the twenty-four APEC isolates, six (25%) isolates had four virulence genes, fourteen (58.33%) isolates carried at least three virulence genes, three (12.5%) isolates carried two genes and one (4.16%) isolates had one virulence gene. Most importantly. six types of virulence gene profiles existed within the APEC isolates from which profile number 3 (hlyF, iroN, iss) having 13 (54.16%) isolates were predominant. The occurrence of APEC isolates of this region which is responsible for avian colibacillosis cases can be a matter of concern from the public health point of view. Future investigations will be able to utilize these virulence genes to identify APEC in Bangladesh helping in the diagnosis and prevention of colibacillosis in poultry. Bioresearch Commu. 7(1): 967-972, 2021 (January)

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Exposure to Bacillus cereus in Water Buffalo Mozzarella Cheese

Foods ◽

10.3390/foods9121899 ◽

2020 ◽

Vol 9 (12) ◽

pp. 1899

Author(s):

Angela Michela Immacolata Montone ◽

Federico Capuano ◽

Andrea Mancusi ◽

Orlandina Di Maro ◽

Maria Francesca Peruzy ◽

...

Keyword(s):

Bacillus Cereus ◽

Dairy Products ◽

Water Buffalo ◽

Diarrheal Disease ◽

Food Poisoning ◽

Mozzarella Cheese ◽

Food Borne ◽

Genes Encoding ◽

Spoilage Bacterium ◽

Hemolysin Bl

Bacillus cereus is a spoilage bacterium and is recognized as an agent of food poisoning. Two food-borne illnesses are caused by B. cereus: a diarrheal disease, associated with cytotoxin K, hemolysin BL, non-hemolytic enterotoxin and enterotoxin FM, and an emetic syndrome, associated with the cereulide toxin. Owing to the heat resistance of B. cereus and its ability to grow in milk, this organism should be considered potentially hazardous in dairy products. The present study assessed the risk of B. cereus poisoning due to the consumption of water buffalo mozzarella cheese. A total of 340 samples were analyzed to determine B. cereus counts (ISO 7932:2005); isolates underwent molecular characterization to detect the presence of genes encoding toxins. Eighty-nine (26.1%) samples harbored B. cereus strains, with values ranging from 2.2 × 102 to 2.6 × 106 CFU/g. Isolates showed eight different molecular profiles, and some displayed virulence characteristics. Bacterial counts and the toxin profiles of isolates were evaluated both separately and jointly to assess the risk of enteritis due to B. cereus following the consumption of buffalo mozzarella cheese. In conclusion, the results of the present study showed that the risk of poisoning by B. cereus following the consumption of this cheese was moderate.

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