Comparison of TEMPO® BC with Spiral Plating Methods for the Enumeration of Bacillus cereus in Cosmetic Products Either Naturally Preserved or Preserved with Phenoxyethanol

2019 ◽  
Vol 102 (4) ◽  
pp. 1080-1090
Author(s):  
Nadine Yossa ◽  
James Smiley ◽  
Mei-Chiung Jo Huang ◽  
Lanlan Yin ◽  
Rebecca Bell ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Number System ◽  
Most Probable Number ◽  
Organic Substances ◽  
Cosmetic Products ◽  
Probable Number ◽  
Liquid Products ◽  
Group Members ◽  
Powder Type ◽  
The U.S

Abstract Background: The U.S. Food and Drug Administration’s (FDA) Bacteriological Analytical Manual (BAM) uses Bacillus cereus rapid agar (BACARA) and Mannitol-yolk-polymyxin (MYP) agar for the enumeration of the members of B. cereus group. Objective: The automated TEMPO Most Probable Number system was compared with the FDA BAM method for the detection of B. cereus group members in cosmetic products. Methods: We inoculated a range of cosmetic products with pure B. cereus spore suspensions (density = 0.5 McFarland) at high (6 log CFU/mL), medium (5 log CFU/mL), and low (4 log CFU/mL) levels. Test portions were aged for 72 h. Five replicates per sample were analyzed; uninoculated test portions served as controls. We also evaluated whether TEMPO BC erroneously detected non-B. cereus or other adulterant organisms. Results: No significant differences (P > 0.05) were found among the TEMPO BC and the BAM spiral plating methods. Correlations between TEMPO BC – BACARA and TEMPO BC – MYP were 0.895 and 0.893 for powder type products, 0.834 and 0.846 for cream and oil-based products, and 0.929 and 0.923 for liquid products, respectively. Non-B. cereus strains were not detected by TEMPO BC. Conclusions: The TEMPO BC method can be used for the detection of B. cereus in cosmetic products without preservatives, or those preserved with either phenoxyethanol or other organic substances.

Comparison of Different Culture Methods for the Detection of Bacillus cereus Group in Cosmetics

2020 ◽  
Vol 103 (4) ◽  
pp. 1129-1139
Author(s):  
Nadine Yossa ◽  
Son T Hoang ◽  
Travis Canida ◽  
Rebecca Bell ◽  
Sandra Tallent ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Relative Effectiveness ◽  
Tween 80 ◽  
Culture Method ◽  
Cosmetic Products ◽  
Culture Methods ◽  
Aging Period ◽  
Liquid Products ◽  
Group Members ◽  
Microbiological Analyses

Abstract Background The U.S. Food and Drug Administration (FDA) Bacteriological Analytical Manual (BAM) reference culture method uses Modified Letheen Broth (MLB) for microbiological analyses for all types of cosmetic products. Objective This study evaluated the effectiveness of MLB and Tryptone Azolectin Tween (TAT) broths using BAM reference culture method for cosmetics. Methods Pure spore suspensions of B. cereus group members were experimentally spiked (McF: 0.5) into cosmetic products. After an aging period of 72 h, the products were analyzed using MLB and TAT broth. The enumeration of the cells was performed on B. cereus group selective plates Bacillus cereus rapid agar (BACARA) and Mannitol Yolk Polymyxin (MYP) plates. Results No statistical difference (p > 0.05) was found for the recovery of cells from the liquid products using either medium (MLB or TAT broth) and the selective plates. In solid/powder products, a combination of Tween 80 and MLB detected significantly more cells (p < 0.05) than combination of Tween 80 and TAT broth. The microbial counts on BACARA showed no significant differences (p > 0.05). However, when assessing cream/oil-based products, the number of cells detected by use of Tween 80/TAT broth was significantly higher than Tween 80/MLB, and MYP showed significantly higher counts than BACARA. Conclusions This study showed that relative effectiveness of MLB vs. TAT for recovering of B. cereus group cells varied depending on the variety of formulation, and combination of preservatives of the tested cosmetic products. The findings suggest additional studies are needed to explore recovery of other relevant microorganisms that may contaminate cream/oil-based cosmetics.

Development of Rapid Real-Time PCR and Most-Probable-Number Real-Time PCR Assays To Quantify Enterotoxigenic Strains of the Species in the Bacillus cereus Group

2007 ◽  
Vol 70 (12) ◽  
pp. 2774-2781 ◽  
Author(s):  
I-CHEN YANG ◽  
DANIEL YANG-CHIH SHIH ◽  
JAN-YI WANG ◽  
TZU-MING PAN
Keyword(s):  
Bacillus Cereus ◽  
Real Time ◽  
Real Time Pcr ◽  
Food Samples ◽  
Most Probable Number ◽  
Pcr Assay ◽  
Bacillus Cereus Group ◽  
Probable Number ◽  
Cooked Rice ◽  
Group Detection

Members of the Bacillus cereus group may produce diarrheal enterotoxins and could be potential hazards if they enter the food chain. Therefore, a method capable of detecting all the species in the B. cereus group rather than B. cereus alone is important. We selected nhe as the target and developed a real-time PCR assay to quantify enterotoxigenic strains of the B. cereus group. The real-time PCR assay was evaluated with 60 B. cereus group strains and 28 others. The assay was also used to construct calibration curves for different food matrices and feces. The assay has an excellent quantification capacity, as proved by its linearity (R2 > 0.993), wide dynamic quantification range (102 to 107 CFU/g for cooked rice and chicken, 103 to 107 CFU/ml for milk, and 104 to 107 CFU/g for feces), and adequate relative accuracy (85.5 to 101.1%). For the low-level contaminations, a most-probable-number real-time PCR assay was developed that could detect as low as 100 CFU/ml. Both assays were tested with real food samples and shown to be considerably appropriate for B. cereus group detection and quantification.

scholarly journals Methods for the Recovery of Salmonella spp. from Carboxymethylcellulose Gum, Gum Ghatti, and Gelatin

1998 ◽  
Vol 81 (4) ◽  
pp. 721-726 ◽  
Author(s):  
René Miguel Amaguaña ◽  
Thomas S Hammack ◽  
Wallace H Andrews
Keyword(s):  
Final Concentration ◽  
Inorganic Salts ◽  
Most Probable Number ◽  
Salmonella Spp ◽  
Test Portion ◽  
Probable Number ◽  
Ph Adjustment ◽  
Gum Ghatti ◽  
Thickening Agents ◽  
The U.S

Abstract Foods analyzed for Salmonella spp. by the procedure in the U.S. Food and Drug Administration's Bacteriological Analytical Manual are preenriched at a 1:9 test portion/broth ratio. Various thickening agents preenriched at this ratio become viscous and nonpipettable after 24 h incubation at 35 C. The effects of 3 factors (presence of inorganic salts, adjustment of pH, and presence of enzymes) on the viscosity of the test portion/preenrichment mixtures of various thickening agents during incubation were determined. Reduction of the viscosities of these thickening agents was accomplished as follows: carboxymethylcellulose gum, addition of cellulase to a final concentration of 0.10℅ in lactose broth preenrichment and incubation with no pH adjustment; gum ghatti, addition of NaCI to a final concentration of 0.10℅ in lactose broth preerichment and adjustment of the pH to 6.5; and gelatin, addition of papain to a final concentration of 0.10℅ in lactose broth preenrichment and adjustment of the pH to 6.8. With these modified preenrichments, one Salmonella spp. cell/25 g (representing an approximate most probable number value of 0.04 cell/g) was generally recovered from the thickening agents.

scholarly journals Prevalence of Bacillus cereus s.l. in ultra-high temperature chocolate milk from selected milk manufacturers in Malaysia

Food Research ◽  
2020 ◽  
Vol 4 (4) ◽  
pp. 982-990
Author(s):  
Ubong Anyi ◽  
C.Y. New ◽  
L.C. Chai ◽  
Y.Y. Loo ◽  
Nor Khaizura M.A.R. ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Foodborne Pathogen ◽  
Most Probable Number ◽  
Contamination Level ◽  
Chocolate Milk ◽  
Probable Number ◽  
Potential Source ◽  
Milk Contamination ◽  
Bacillus Spp ◽  
Polymerase Chain

Bacillus cereus is a major foodborne pathogen of great concern to the dairy industry owing to its resilient spores as well as the adverse effect of its toxins. At present, there is no informational study available to solve or pinpoint the UHT chocolate milk contamination issue in Malaysia. This work aimed to investigate the prevalence and contamination level of B. cereus s.l. in UHT chocolate milk and to suggest the appropriate solution for the issue. In the present study, B. cereus s.l. prevalence and contamination level in individually packed UHT chocolate milk from processing factories was evaluated. The prevalence and concentration of B. cereus s.l. were determined via MPN-PCR (Most Probable Number-Polymerase Chain Reaction) assay. Results showed that 31.11% from 220 of UHT chocolate milk tested contained Bacillus spp.; of this Bacillus spp. positive samples, 24.30% were also positive for B. cereus s.l. with concentration ranging from less than 3 to more than 1100 MPN/mL. Findings from this study highlighted the possibility of UHT chocolate milk as a potential source of B. cereus s.l. infection. Therefore, findings emphasized the needs to revise, monitor and improve UHT sterilization process to reduce infection risk. Furthermore, it is also essential to maintain the hygiene to minimize initial microbial load and contamination of UHT chocolate milk, beginning from production to retail.

Precision Parameters for the FDA Shellfish Laboratory Quality Assurance Program for 1973–1989

1995 ◽  
Vol 58 (6) ◽  
pp. 648-650
Author(s):  
JAMES T. PEELER ◽  
THOMAS E. GRAHAM ◽  
LARRY J. MATURIN
Keyword(s):  
Staphylococcus Aureus ◽  
Quality Assurance ◽  
Fecal Coliform ◽  
Quality Assurance Program ◽  
Plate Count ◽  
Most Probable Number ◽  
Probable Number ◽  
Standard Plate Count ◽  
Standard Plate ◽  
The U.S

Precision parameters from four microbiological analytical methods (coliform most probable number [MPN], fecal coliform MPN, Staphylococcus aureus plate count and standard plate count) were computed for the Shellfish Quality Assurance Program of the U.S. Food and Drug Administration (FDA). The pooled reproducibility variance (SR2) for the four methods from 1973 to 1989 were 0.0778, 0.1181, 0.0137, and 0.0087, respectively.

scholarly journals Microbial Communities in Contaminated Sediments, Associated with Bioremediation of Uranium to Submicromolar Levels

2008 ◽  
Vol 74 (12) ◽  
pp. 3718-3729 ◽  
Author(s):  
Erick Cardenas ◽  
Wei-Min Wu ◽  
Mary Beth Leigh ◽  
Jack Carley ◽  
Sue Carroll ◽  
...  
Keyword(s):  
Microbial Communities ◽  
Contaminated Site ◽  
Most Probable Number ◽  
Biological Reduction ◽  
Probable Number ◽  
Oak Ridge ◽  
Sulfate Reducing ◽  
Treatment Zone ◽  
The U.S

ABSTRACT Microbial enumeration, 16S rRNA gene clone libraries, and chemical analysis were used to evaluate the in situ biological reduction and immobilization of uranium(VI) in a long-term experiment (more than 2 years) conducted at a highly uranium-contaminated site (up to 60 mg/liter and 800 mg/kg solids) of the U.S. Department of Energy in Oak Ridge, TN. Bioreduction was achieved by conditioning groundwater above ground and then stimulating growth of denitrifying, Fe(III)-reducing, and sulfate-reducing bacteria in situ through weekly injection of ethanol into the subsurface. After nearly 2 years of intermittent injection of ethanol, aqueous U levels fell below the U.S. Environmental Protection Agency maximum contaminant level for drinking water and groundwater (<30 μg/liter or 0.126 μM). Sediment microbial communities from the treatment zone were compared with those from a control well without biostimulation. Most-probable-number estimations indicated that microorganisms implicated in bioremediation accumulated in the sediments of the treatment zone but were either absent or in very low numbers in an untreated control area. Organisms belonging to genera known to include U(VI) reducers were detected, including Desulfovibrio, Geobacter, Anaeromyxobacter, Desulfosporosinus, and Acidovorax spp. The predominant sulfate-reducing bacterial species were Desulfovibrio spp., while the iron reducers were represented by Ferribacterium spp. and Geothrix spp. Diversity-based clustering revealed differences between treated and untreated zones and also within samples of the treated area. Spatial differences in community structure within the treatment zone were likely related to the hydraulic pathway and to electron donor metabolism during biostimulation.

scholarly journals Development of a Fluorogenic Probe-Based PCR Assay for Detection of Bacillus cereus in Nonfat Dry Milk

2000 ◽  
Vol 66 (4) ◽  
pp. 1453-1459 ◽  
Author(s):  
Young-Rok Kim ◽  
John Czajka ◽  
Carl A. Batt
Keyword(s):  
Bacillus Cereus ◽  
Positive Sequence ◽  
Most Probable Number ◽  
Pcr Assay ◽  
Probable Number ◽  
Fluorogenic Probe ◽  
Hemolysin Gene ◽  
Dry Milk ◽  
Fluorogenic Probes ◽  
Nonfat Dry Milk

ABSTRACT A fluorogenic probe-based PCR assay was developed and evaluated for its utility in detecting Bacillus cereus in nonfat dry milk. Regions of the hemolysin and cereolysin AB genes from an initial group of two B. cereus isolates and two Bacillus thuringiensis isolates were cloned and sequenced. Three single-base differences in two B. cereus strains were identified in the cereolysin AB gene at nucleotides 866, 875, and 1287, while there were no species-consistent differences found in the hemolysin gene. A fluorogenic probe-based PCR assay was developed which utilizes the 5′-to-3′ exonuclease of Taq polymerase, and two fluorogenic probes were evaluated. One fluorogenic probe (cerTAQ-1) was designed to be specific for the nucleotide differences at bases 866 and 875 found in B. cereus. A total of 51 out of 72B. cereus strains tested positive with the cerTAQ-1 probe, while only 1 out of 5 B. thuringiensis strains tested positive. Sequence analysis of the negative B. cereusstrains revealed additional polymorphism found in the cereolysin probe target. A second probe (cerTAQ-2) was designed to account for additional polymorphic sequences found in the cerTAQ-1-negativeB. cereus strains. A total of 35 out of 39 B. cereus strains tested positive (including 10 of 14 previously negative strains) with cerTAQ-2, although the assay readout was uniformly lower with this probe than with cerTAQ-1. A PCR assay using cerTAQ-1 was able to detect approximately 58 B. cereus CFU in 1 g of artificially contaminated nonfat dry milk. Forty-three nonfat dry milk samples were tested for the presence of B. cereus with the most-probable-number technique and the fluorogenic PCR assay. Twelve of the 43 samples were contaminated withB. cereus at levels greater than or equal to 43 CFU/g, and all 12 of these samples tested positive with the fluorogenic PCR assay. Of the remaining 31 samples, 12 were B. cereus negative and 19 were contaminated with B. cereus at levels ranging from 3 to 9 CFU/g. All 31 of these samples were negative in the fluorogenic PCR assay. Although not totally inclusive, the PCR-based assay with cerTAQ-1 is able to specifically detect B. cereus in nonfat dry milk.

scholarly journals Analisis Cemaran Escherichia coli, Salmonella sp., Staphylococcus aureus, dan Bacillus cereus pada Minuman Susu Kedelai di Kota Padang

2019 ◽  
Vol 11 (02) ◽  
pp. 1
Author(s):  
Sari Mustika ◽  
Ranggi Rahimul Insan ◽  
Anni Faridah
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Bacillus Cereus ◽  
Most Probable Number ◽  
Probable Number

Minuman susu kedelai dibuat dengan bahan dasar kedelai digiling halus, ditambahkan air dan disaring, yang kemudian menghasilkan cairan berwarna putih seperti susu. Apabila pada proses pembuatannya tidak baik dan bersih, maka dapat terkontaminasi oleh beberapa bakteri pathogen seperti; bakteri Escherichia Coli, Salmonella sp. sp., Staphylococcus aureus, dan Bacillus cereus yang dapat menyebabkan timbulnya berbagai penyakit pada orang yang mengkonsumsi. Penelitian ini merupakan penelitian deskriptif dengan metode pendekatan kualitatif, bertujuan untuk menganalisis dan mengetahui adanya cemaran bakteri patogen Escherichia Coli, Salmonella sp, Staphylococcus aureus, dan Bacillus cereus yang terdapat pada minuman jajanan susu kedelai di Kota Padang. Penelitian ini menggunakan sampel sebanyak 9 buah yang terdiri dari minuman susu kedelai yang didapat dari pasar tradisional di Kota Padang. Jumlah bakteri pada masing-masing sampel dihitung menggunakan metode Most Probable Number (MPN). Hasil penelitian diperoleh dari 9 sampel yang diuji sebagian besar sampel positif tercemar bakteri patogen dan jumlahnya melebihi dari batas yang sesuai yang syaratkan SNI 7388-2009 tentang syarat mutu minuman susu kedelai.

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