• Title/Summary/Keyword: Bacillus cereus group

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Riboprint and Virulence Gene Patterns for Bacillus cereus and Related Species

  • Kim, Young-Rok;Batt, Carl A.
    • Journal of Microbiology and Biotechnology
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    • v.18 no.6
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    • pp.1146-1155
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    • 2008
  • A total of 72 Bacillus cereus strains and 5 Bacillus thuringiensis strains were analyzed for their EcoRI ribogroup by ribotyping and for the presence or absence of seven virulence-associated genes. From these 77 strains, 42 distinctive ribogroup were identified using EcoRI, but the two species could not be discriminated by their EcoRI ribogroup. The 77 strains were also examined by PCR for the presence of seven virulence-associated genes, cerAB, pi-plc, entFM, bceT, hblA, hblC, and hblD. All five Bacillus thuringiensis strains were positive for these genes. Although differences in the patterns of virulence genes were observed among the different B. cereus strains, within any given ribogroup the patterns of the seven virulence genes was the same. Pulsed-field gel electrophoresis (PFGE) analysis in combination with available chromosomal maps for a selected group of B. cereus strains revealed significant differences in their chromosome size and the placement of virulence genes. Evidence for significant rearrangements within the B. cereus chromosome suggests the mechanism through which the pattern of virulence-associated genes varies. The results suggest linkage between ribogroups and virulence gene patterns as well as no apparent containment of the latter within any particular species boundary.

Antibacterial and virucidal activity of 28 extracts from plants endemic to Korea against Bacillus cereus, Staphylococcus aureus, and murine norovirus

  • Hyun-Joo Chang;Ji Hye Han;Nari Lee;Sung-Wook Choi
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.345-352
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    • 2023
  • Antibacterial activity against foodborne bacteria (Bacillus cereus, Staphylococcus aureus, Salmonella Enteritidis) and inhibitory activity against murine norovirus, a human norovirus surrogate, of 28 extracts from plants endemic to Korea were investigated in this study. All plant extracts showed antibacterial activity only against gram-positive bacteria, B. cereus and S. aureus. Extracts from Callistemon speciosus and Nymphaea tetragona showed inhibition zones of 16.54 and 24.35 mm against B. cereus and S. aureus, respectively, presenting the highest antibacterial activities recorded in this study. Among all samples, Ardisia japonica extract at concentrations of 100 and 200 ㎍/mL showed the highest virucidal activities of 96.6 and 100.0%, respectively. Ardisia japonica, Duchesnea indica, Polygonum aviculare, and Geum japonicum extracts showed high antibacterial and virucidal activity simultaneously without Raw 264.7 cell cytotoxicity. These plant extracts may serve as potential antimicrobials to control foodborne infections.

Reliable Identification of Bacillus cereus Group Species Using Low Mass Biomarkers by MALDI-TOF MS

  • Ha, Miyoung;Jo, Hyeon-Ju;Choi, Eun-Kyeong;Kim, Yangsun;Kim, Junsung;Cho, Hyeon-Jong
    • Journal of Microbiology and Biotechnology
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    • v.29 no.6
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    • pp.887-896
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    • 2019
  • Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)-based pathogen identification relies on the ribosomal protein spectra provided in the proprietary database. Although these mass spectra can discern various pathogens at species level, the spectra-based method still has limitations in identifying closely-related microbial species. In this study, to overcome the limits of the current MALDI-TOF MS identification method using ribosomal protein spectra, we applied MALDI-TOF MS of low-mass profiling to the identification of two genetically related Bacillus species, the food-borne pathogen Bacillus cereus, and the insect pathogen Bacillus thuringiensis. The mass spectra of small molecules from 17 type strains of two bacilli were compared to the morphological, biochemical, and genetic identification methods of pathogens. The specific mass peaks in the low-mass range (m/z 500-3,000) successfully identified various closely-related strains belonging to these two reference species. The intensity profiles of the MALDI-TOF mass spectra clearly revealed the differences between the two genetically-related species at strain level. We suggest that small molecules with low molecular weight, 714.2 and 906.5 m/z can be potential mass biomarkers used for reliable identification of B. cereus and B. thuringiensis.

Biotransformation of Flavonoids with O-Methyltransferase from Bacillus cereus

  • Lee Yoon-Jung;Kim Bong-Gyu;Park Young-Hee;Lim Yoong-Ho;Hur Hor-Gil;Ahn Joong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.16 no.7
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    • pp.1090-1096
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    • 2006
  • O-Methylation is a common modification reaction found in nature, and is mediated by an O-methyltransferase (OMT). OMTs have been mainly studied in plants, whereas only a few OMTs have been studied in microbes. When searching the Bacillus cereus genome, four putative small molecular OMTs were identified, among which BcOMT-1 was cloned and expressed in E. coli as a his-tag fusion protein. The whole cell expressing BcOMT-1 was used to methylate several flavonoids. Eriodictyol, luteolin, quercetin, and taxifolin, all of which contain 3' and 4' hydroxyl groups, served as methyl group acceptors for BcOMT-1, whereas naringenin, apigenin, 3,3'-dihydroxyflavone, and 3,4'-dihydroxyflavone did not function as substrates. Analysis of the reaction products using HPLC showed two different peaks, and NMR revealed that the methylation position was at the hydroxyl group of either carbon 3' or 4'. Therefore, this showed that BcOMT-1 used flavonoids containing ortho hydroxyl groups and transferred a methyl group to either of two hydroxyl groups.

Impact of the Isolation Source on the Biofilm Formation Characteristics of Bacillus cereus

  • Hussain, Mohammad Shakhawat;Oh, Deog-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.28 no.1
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    • pp.77-86
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    • 2018
  • The human pathogen and food spoiler Bacillus cereus can form biofilms that act as a persistent source of contamination, which is of public health concern. This study aimed to understand how the source of isolation might affect the behavior of biofilm formation. Biofilm formation abilities of 56 strains of B. cereus isolated from different environments, including human food poisoning, farm, and food, were determined. Crystal violet assay results revealed significant (p < 0.05) differences in biofilm formation abilities among the strains isolated from different sources only at an early stage of incubation. However, strain origin showed no impact on later stage of biofilm formation. Next, correlation of the group of isolates on the basis of their biofilm-forming abilities with the number of sessile cells, sporulation, and extracellular polymeric substance (EPS) formation was determined. The number of sessile cells and spores in biofilms was greatly influenced by the groups of isolates that formed dense, moderate, and weak biofilms. The contribution of extracellular DNA and/or proteins to EPS formation was also positively correlated with biofilm formation abilities. Our results that the source of isolation had significant impact on biofilm formation might provide important information to develop strategies to control B. cereus biofilm formation.

Generation of a Specific Marker to Discriminate Bacillus anthracis from Other Bacteria of the Bacillus cereus Group

  • Kim, Hyoung-Tai;Seo, Gwi-Moon;Jung, Kyoung-Hwa;Kim, Seong-Joo;Kim, Jee-Cheon;Oh, Kwang-Geun;Koo, Bon-Sung;Chai, Young-Gyu
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.806-811
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    • 2007
  • Bacillus anthracis is a soil pathogen capable of causing anthrax that is closely related to several environmental species, including B. cereus, B. mycoides, and B. thuringiensis. DNA homology studies showed that B. anthracis, B. cereus, B. mycoides, and B. thuringiensis are closely related, with a high sequence homology. To establish a method to specifically detect B. anthracis in situations such as environmental contamination, we initially performed RAPD-PCR with a 10-mer random primer and confirmed the presence of specific PCR bands only in B. anthracis species. One region specific for B. anthracis was cloned and sequenced, and an internal primer set was designed to amplify a 241-bp DNA fragment within the sequenced region. The PCR system involving these specific primer sets has practical applications. Using lyses methods to prepare the samples for PCR, it was possible to quickly amplify the 241-bp DNA segment from samples containing only a few bacteria. Thus, the PCR detection method developed in this study is expected to facilitate the monitoring of environmental B. anthracis contamination.

Survival, isolation and characterization of Bacillus cereus from Sunshik (선식에서 Bacillus cereus의 분리 및 특성 조사와 열에 대한 사멸률 연구)

  • Cho, Yong-Sun;Jung, Eun-Young;Lee, Myung-Ki;Yang, Cheul-Young;Shin, Dong-Bin
    • Journal of Food Hygiene and Safety
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    • v.23 no.4
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    • pp.343-347
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    • 2008
  • Recently, Sunshik has been issued because of easy-cook and well-being food. Sunshik basically was made of the heated cereals. Amount of spore-forming Bacillus cereus was detected and it has been caused some problem of food safety. B. cereus was isolate from 57 out of 161 Sunshik samples resulting in the isolation rate of 35.4%. Quantitative analysis of 57 samples showed that 21 samples were less than 100 CFU/g, 33 samples were between 100 and 1,000 CFU/g and distinctively even 3 (1.9%) samples had over 1,000 CFU/g. Typical morphology of B. cereus isolated from Sunshik was observed on MYP agar and then further characteristics was identified by using VITEK 2 (Biomeriux, France). 53 strains out of 57 strains isolated from Sunshik (about 93.0%) produced diarrheal enterotoxin in brain heart infusion broth which was detected by the Bacillus cereus enterotoxin reversed passive latex agglutination test kit (Oxoid England). The D-values of the B. cereus spores were $75^{\circ}C$ (37.1mim), $80^{\circ}C$ (22.5mim), $85^{\circ}C$ (4.9mim), and $90^{\circ}C$ (3.1mim) respectively. The Z-value was calculated $12.8^{\circ}C$ in Sunshik sample inoculated with B. cereus. Therefore, the management of B. cereus in Sunshik is required for the food-safety.

Effect of LED light on the inactivation of Bacillus cereus for extending shelf-life of extruded rice cake and simulation of the patterns of LED irradiation by various arrays of LEDs (압출떡의 유통기한 연장을 위한 LED 조사의 Bacillus cereus 억제 효과 및 LED의 배열에 따른 빛의 조사 패턴 시뮬레이션)

  • Jung, Hwabin;Yuk, Hyun-Gyun;Yoon, Won Byong
    • Journal of Applied Biological Chemistry
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    • v.62 no.2
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    • pp.181-186
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    • 2019
  • The optimum design of LED device for irradiation of 460 nm blue light on extruded rice cake using simulation and the effect of the blue light on the inactivation of Bacillus cereus (B. cereus) group on the rice cake were investigated. The irradiated light intensity patterns on the surface area of the sample were simulated with three different LED arrays (centered, cross, and evenly spaced) and at various distances (22, 32, 42 mm) between the LED modules and the sample. In addition, the uniformity was calculated as Petri factor. The evenly spaced array resulted the most uniform light intensity pattern in the simulation, and the Petri factor of 32 and 42 mm of the distances showed higher than 0.9, which represents the ideal uniformity of LED device. The bacterial population of the rice cake decreased to less than the initial bacterial population during exposure to LED blue light, whereas the bacterial population of the control sample increased. The bacterial count of the rice cake after blue light irradiation for 24 h was 1.21 log CFU/g lower than the control sample. Petri factor increased with increase of the distance between the light source and sample, however, the reduction rate of B. cereus group decreased. Therefore, the design of LED device, that represented the Petri factor higher than 0.9 and inactivated the population of B. cereus group, with evenly spaced and 32 mm of distance between the light source and sample was suitable for extending shelf-life of rice cake.

Microbial Contamination Analysis to Assess the Safety of Marketplace Sushi (유통중인 생선초밥의 오염 미생물 분석)

  • Cho, Sun-Kyung;Moon, Bo-Youn;Park, Jong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.41 no.3
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    • pp.334-338
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    • 2009
  • To determine the contamination status of Sushi fish and rice, seventy-nine samples of Sushi were collected from wholesale markets and Japanese restaurants within the Seoul area and subsequently analyzed for food-borne pathogens. Total aerobic counts ranged from 4 to 6 log CFU/g for the sliced raw fish, and from 3 to 5 log CFU/g for the boiled rice. Higher levels of contamination were detected in bream and shrimp Sushi versus other types. Coliform counts of 3-4 log CFU/g were detected in the sliced raw fish, whereas levels in the boiled rice were one log CFU/g lower compared to the raw fish. The raw Sushi fish had higher amounts of contamination than the boiled rice, however, E.coli was not detected. The prevalence rates of pathogens, namely Staphylococcus aureus and Bacillus cereus, in the raw fish were 17% and 10%, respectively. Similarly, the prevalence rates in the boiled rice were 11% and 8% for S. aureus and B.cereus, respectively. Salmonella and Listeria monocytogenes were also detected; however, other pathogens such as Vibrio parahaemolyticus, Clostridium perfrigens, and Yersinia enterocolitica were not detected. Among the high contaminating pathogens, B.cereus was found in 13% of samples from the wholesale markets, while S.aureus was found in 30% of samples from the Japanese restaurants. Therefore, these data suggest that the primary microbial hazard factors for Sushi are S. aureus and B. cereus, in addition to V. parahaemolyticus, and further risk assessments should focus on those pathogens.