An improved selective and diagnostic medium for the isolation and enumeration of Bacillus cereus in foods

1980 ◽  
Vol 26 (7) ◽  
pp. 753-759 ◽  
Author(s):  
R. Holbrook ◽  
Judith M. Anderson

The use and performance of an improved diagnostic and selective medium, PEMBA (polymyxin pyruvate egg yolk mannitol bromothymol blue agar), for the detection of Bacillus cereus in foods is described. The distinct colonial appearance of B. cereus on PEMBA permitted the recognition of both strains: those that do precipitate egg yolk and those that do not react with egg yolk. A staining procedure, used to demonstrate microscopically both the presence of lipid globules in vegetative cells and spore morphology of isolates, proved a rapid and reliable confirmatory test which gave complete agreement with a battery of biochemical tests used for this purpose. The quantitative recovery of B. cereus on PEMBA from 143 food samples was not significantly different from counts on KG (Kim and Goepfert), MYP (mannitol egg yolk phenol red), and McClung's media, and the selectivity of PEMBA was generally superior.

1971 ◽  
Vol 34 (1) ◽  
pp. 12-15 ◽  
Author(s):  
H. U. Kim ◽  
J. M. Goepfert

One hundred seventy samples of dried food products in national distribution were examined for the incidence and level of contamination by Bacillus cereus. Twenty-five per cent of the samples yielded B. cereus at a level not exceeding 4000 per gram. Mannitol-egg yolk-polymyxin (MYP) agar was used as a presumptive test for the presence of B. cereus in the food samples. Various biochemical tests for the confirmation of suspicious colonies appearing on MYP agar were evaluated. A precipitin test employing spore precipitinogens was investigated as a confirmatory test for B. cereus. The possible role of B. cereus in outbreaks of foodborne disease in the United States is discussed.


2020 ◽  
Vol 4 (2) ◽  
pp. 68-72
Author(s):  
Marwan Msarah ◽  
Ahmed Alsier ◽  
Sahilah, A.M.

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.


2017 ◽  
Vol 80 (3) ◽  
pp. 502-505
Author(s):  
Il-Byeong Kang ◽  
Jung-Whan Chon ◽  
Dong-Hyeon Kim ◽  
Dana Jeong ◽  
Hong-Seok Kim ◽  
...  

ABSTRACT A modified polymyxin–egg yolk–mannitol–bromothymol blue agar (mPEMBA) was developed by supplementing polymyxin–egg yolk–mannitol–bromothymol blue agar (PEMBA) with trimethoprim to improve the selectivity for and recoverability of Bacillus cereus from naturally and artificially contaminated food samples. The number of B. cereus in mPEMBA was significantly higher than in PEMBA, indicating better recoverability (P < 0.05) in red pepper powder (PEMBA 0.80 ± 0.22 log CFU/g versus mPEMBA 1.95 ± 0.17 log CFU/g) and soybean paste (PEMBA 2.19 ± 0.18 log CFU/g versus mPEMBA 3.09 ± 0.13 log CFU/g). In addition, mPEMBA provided better visual differentiation of B. cereus colonies than PEMBA, which is attributable to the reduced number of competing microflora. We conclude that the addition of trimethoprim to PEMBA could generate a synergistic effect to improve selectivity for B. cereus.


2019 ◽  
Vol 7 (1) ◽  
pp. 55-65
Author(s):  
Yulianto Ade Prasetya

Methicillin-resistant Staphyloccocus aureus (MRSA) is an isolate that is resistant to the antibiotic methicillin and beta lactam group. The incidence of MRSA associated with nosocomial infections in various parts of the world is very high, but research on its spread in community infections is rarely reported. This study aims to detect the presence of phenotypic MRSA in food samples in Sidoarjo. The food samples (cilok, fried foods and tempura) collected were then weighed, diluted, and cultured in a selective medium and differential namely Manitol Salt agar. The yellow-colored colonies were then continued with microscopic testing and biochemical tests to distinguish between Staphylococcus species. Thirty eight collected Staphyloccus aureus isolates were then screened using Oxacillin 1 µg and there were eight (8) isolates that were positive for MRSA according to the criteria of the Clinical Laboratory Standart Institutre (CLSI). Eight of the isolates were tested for antibiotic sensitivity with the Kirby-Bauer method with Chloramphenicol 30 µg and Cotrimoxazole 25 µg. Eight MRSA (21%) isolates were resistant to Chloramphenicol and only four isolates were resistant to Cotrimoxazole. The presence of MRSA isolates in community infections needs to be watched out for considering these genes can be transmitted and spread between bacterial species


1982 ◽  
Vol 45 (3) ◽  
pp. 218-222 ◽  
Author(s):  
LEONIE MINTZER-MORGENSTERN ◽  
ELIY AHU KATZENELSON

An agar medium containing NaCl, egg yolk and tellurite for selective quantitative isolation of coagulase-positive staphylococci from food was developed. Isolation and identification of the staphylococci was achieved in a single step. A properly diluted food sample was spread over the medium and incubated for 24 h at 42 C. Coagulase-positive staphylococci appeared as small grey to dark-grey colonies surrounded by a dense white opacity. Coagulase-negative bacteria which, at times, grow on this medium, did not produce this reaction. The identification on this selective medium of isolates from 683 different food samples as coagulase-positive staphylococci was subsequently confirmed by the coagulase test. Comparative titrations of 29 various coagulase-positive staphylococcus strains on both the selective medium and nutrient agar yielded nearly identical titers. The growth of heat-stressed staphylococci was inhibited by the selective medium. Complete reversal of the inhibition was achieved by a 3-h pre-incubation in brain heart infusion at 37 C.


1985 ◽  
Vol 48 (11) ◽  
pp. 969-970 ◽  
Author(s):  
MATS PETERZ ◽  
CHRISTER WIBERG ◽  
PER NORBERG

Three media for isolation of Bacillus cereus from foods were compared: mannitol-egg yolk-polymyxin (MYP) agar, polymyxin pyruvate-egg yolk-mannitol-bromothymol blue agar (PEMBA) and non-selective blood agar. Twenty-six of 45 samples of different reconstituted and incubated dry food products and 18 of 29 samples of milk and cream (incubated overnight) contained B. cereus. None of the media performed significantly better than the others as regards quantitative recovery or selectivity.


Plant Disease ◽  
2014 ◽  
Vol 98 (7) ◽  
pp. 988-988 ◽  
Author(s):  
C. Riera-Ruiz ◽  
J. Vargas ◽  
C. Cedeño ◽  
P. Quirola ◽  
M. Escobar ◽  
...  

Rice (Oryza sativa L.) is one of the leading crops and the basis of most diets in Ecuador and other countries. Diseases such as bacterial panicle blight (BPB), also known as seedling rot or grain rot, have the potential to threaten rice production worldwide. Burkholderia glumae, a causal agent of BPB, has severely affected the rice industry in many countries of Africa, Asia, and the Americas (1,2,4), but no report of this bacteria in Ecuador can be found in the literature. Rice plantations showing BPB-like symptoms including upright panicles with stained and vain grains were spotted in Palestina city, one of Ecuador's most extensive rice areas, in July 2013, but similar symptoms have been observed in the region since early 2012. Six symptomatic plants from two different groves were collected. Samples were plated on the semi-selective medium S-PG (KH2PO4 1.3 g, Na2HPO4 1.2 g, (NH4)2SO4 5 g, MgSO4·7H2O 0.25 g, Na2MoO4·2H2O 24 mg, EDTA-Fe 10 mg, L-cystine 10 μg, D-sorbitol 10 g, pheneticillin potassium 50 mg, ampicillin sodium 10 mg, cetrimide 10 mg, methyl violet 1 mg, phenol red 20 mg, agar 15 g/liter distilled water) and axenic colonies were transferred to potato dextrose agar (PDA) to test for fluorescence (3). Colonies of the potential pathogen were 1 mm, circular, entire margin, with a smooth and shiny surface. When cultured in PDA, isolates showed a moist texture, dull yellow color, and displayed fluorescence with exposure to UV light. Cells were bacterial gram-negative rods of 1 to 2 × 0.5 μm. Twelve presumptive isolates were submitted to biochemical tests (API 20NE). The biochemical profile (APIWEB) showed that all the isolates belonged to the Burkholderia genus with a 99.9% similarity. To determine the bacterial species, colonies were submitted to ELISA tests using specific antibodies for B. glumae from Agdia, Inc. The two isolates that were positive for B. glumae were sequenced using a part of the 16s rDNA amplified by the primers 536F: 5′-GTGCCAGCMGCCGCGGTAATAC-3′ and 1492R: 5′-GGTTACCTTGTTACGACTT-3′. The obtained sequences (deposited into GenBank as KF601202) shared 100% similarity with several B. glumae strains after a BLAST query. Isolates were then diluted to 108 UFC/ml and used to inoculate healthy rice plants. Inoculated plants produced BPB-like symptoms including upright panicles with stained vain grains and the bacterium was re-isolated from symptomatic plants. To the best of our knowledge, this is the first report of B. glumae in Ecuador. Further research is ongoing to identify and determine the pathogenicity of the remaining Burkholderia strains that tested negative for B. glumae. References: (1) J. Luo et al. Plant Dis. 91:1363, 2007. (2) R. Nandakumar et al. Plant Dis. 93:896, 2009. (3) T. Urakami et al. Int. J. Syst. Bacteriol. 44:235, 1994. (4) X.-G. Zhou. Plant Dis. 98:566, 2014.


2012 ◽  
Vol 95 (2) ◽  
pp. 446-451 ◽  
Author(s):  
Sandra M Tallent ◽  
Kristin M Kotewicz ◽  
Errol A Strain ◽  
Reginald W Bennett

Abstract Bacillus cereus is a group of ubiquitous facultative anaerobic sporeforming Gram-positive rods commonly found in soil. The spores frequently contaminate a variety of foods, including produce, meat, eggs, and dairy products. Foodborne illnesses associated with toxins produced by B. cereus can result in self-limiting diarrhea or vomiting. Plate enumeration methods recommended by recognized food authorities to detect the presence of B. cereus in potentially contaminated food products do not inhibit other Gram-positive competitive bacteria. This study evaluated the use of Bacara, a new chromogenic agar, as an efficient method to identify and enumerate B. cereus group from food matrixes, even in the presence of background flora. Inclusivity and exclusivity testing was performed using four different selective and differential media for B. cereus, including Mannitol Egg Yolk Polymyxin (MYP), Polymyxin Pyruvate Egg-Yolk Mannitol Bromothymol Blue Agar, Bacillus Chromogenic Media, Brilliance, and Bacara. MYP and Bacara were also used in plate enumeration studies to isolate B. cereus from artificially contaminated foods.


2020 ◽  
Vol 70 (2) ◽  
pp. 182-193
Author(s):  
Savić Dejana ◽  
Ristanović Elizabeta ◽  
Miljković Selimović Biljana ◽  
Radaković Sonja ◽  
Jošić Dragana ◽  
...  

AbstractBacillus cereus, usually ingested by food, can cause two types of disease due to the presence of toxins: vomiting and diarrhea syndrome. Systemic infections can also occur. The aim was to detect genes for enterotoxins (hblA, entFM) and emetic toxin (cer) and to investigate the genetic heterogeneity of B. cereus isolates from food, environment and human stool. Identification of B. cereus was performed by means of selective medium, classical biochemical test and polymerase chain reaction (PCR). Toxin genes were detected by PCR. Typing was performed by random amplified polymorphic DNA (RAPD). EntFM gene was present in all stool and food samples and in 28/30 environmental isolates. HblA gene was present in 29/30 stool, 23/30 food and 24/30 environmental isolates. Cer gene was present in 30/30 stool, 28/30 food and 25/30 environmental isolates. The RAPD results show high heterogeneity among the isolates from each group. In the cumulative dendrogram, representative isolates from all three groups formed two clusters with a difference of 53%. The detection of toxin genes in all B. cereus isolates indicated these bacteria as potentially pathogenic and a serious threat for human health. The presence of isolates from all three groups in the same cluster suggests the existence of similar strains in the environment, food and patients, which is in line with the circulation of strains in nature through the food chain.


2018 ◽  
Vol 7 (2) ◽  
pp. 131-136
Author(s):  
Nasir Ahmad

Background: On May 4th, 2016, at 12:30 district surveillance officer of Magelang Health Department received reports from Public Health Center of Bandongan about 21 students of SDN 1 Trasan who suffered from the same food-poisoning symptoms. Objective: Investigation was carried out to identify the source, how it spread and how to control it. Methods: This study used descriptive analytic and mapping the cases distribution location. The case was people experiencing symptoms of dizziness or abdominal pain or nausea or vomiting. Data analysis was done by using bivariate analysis. Data collection were done through interviews, observations and laboratory tests on the food samples. Results: The case was 50 students (from 1-6 grade students). The perceived symptoms were dizziness (77%), nausea (42%), abdominal pain (40%) and vomiting (8%). Attack rate found ranged from 14.3% to 60% with the highest Attack rate found on class three (60%). The incubation period of 15-240 minutes (mean 72.3 minutes). Calamari like positive Bacillus cereus and Rhodamine-B 10 mg/kg. Conclusion: The outbreak of food poisoning because calamari like contaminated Bacillus cereus. We suggested the school committee to provide the socialization of harmful food for the students. The teachers should restrict the permission for the food vendor to sell at school.   Keywords: Bacillus cereus, , Food Poisoning, Outbreak, Rhodamine B, School Food


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