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.

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.

Relative Effectiveness of the Bacteriological Analytical Manual Method for the Recovery of Salmonella from Whole Cantaloupes and Cantaloupe Rinses with Selected Preenrichment Media and Rapid Methods†

2004 ◽  
Vol 67 (5) ◽  
pp. 870-877 ◽  
Author(s):  
THOMAS S. HAMMACK ◽  
IRIS E. VALENTIN-BON ◽  
ANDREW P. JACOBSON ◽  
WALLACE H. ANDREWS
Keyword(s):  
Relative Effectiveness ◽  
Culture Method ◽  
Cell Suspensions ◽  
Culture Methods ◽  
Rapid Methods ◽  
Manual Method ◽  
Plastic Bags ◽  
Peptone Water ◽  
Sterile Plastic

Soak and rinse methods were compared for the recovery of Salmonella from whole cantaloupes. Cantaloupes were surface inoculated with Salmonella cell suspensions and stored for 4 days at 2 to 6° C. Cantaloupes were placed in sterile plastic bags with a nonselective preenrichment broth at a 1:1.5 cantaloupe weight-to-brothvolume ratio. The cantaloupe broths were shaken for 5 min at 100 rpm after which 25-ml aliquots (rinse) were removed from the bags. The 25-ml rinses were preenriched in 225-ml portions of the same uninoculated broth type at 35° C for 24 h (rinse method). The remaining cantaloupe broths were incubated at 35° C for 24 h (soak method). The preenrichment broths used were buffered peptone water (BPW), modified BPW, lactose (LAC) broth, and Universal Preenrichment (UP) broth. The Bacteriological Analytical Manual Salmonella culture method was compared with the following rapid methods: the TECRA Unique Salmonella method, the VIDAS ICS/SLM method, and the VIDAS SLM method. The soak method detected significantly more Salmonella-positive cantaloupes (P < 0.05) than did the rinse method: 367 Salmonella-positive cantaloupes of 540 test cantaloupes by the soak method and 24 Salmonella-positive cantaloupes of 540 test cantaloupes by the rinse method. Overall, BPW, LAC, and UP broths were equivalent for the recovery of Salmonella from cantaloupes. Both the VIDAS ICS/SLM and TECRA Unique Salmonella methods detected significantly fewer Salmonella-positive cantaloupes than did the culture method: the VIDAS ICS/SLM method detected 23 of 50 Salmonella-positive cantaloupes (60 tested) and the TECRA Unique Salmonella method detected 16 of 29 Salmonella-positive cantaloupes (60 tested). The VIDAS SLM and culture methods were equivalent: both methods detected 37 of 37 Salmonella-positive cantaloupes (60 tested).

scholarly journals Isolation of Mycobacterium avium Subsp. Paratuberculosis in the Feces and Tissue of Small Ruminants Using a Non-Automated Liquid Culture Method

Animals ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 20
Author(s):  
Luigi De Grossi ◽  
Davide Santori ◽  
Antonino Barone ◽  
Silvia Abbruzzese ◽  
Matteo Ricchi ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Liquid Culture ◽  
Culture Media ◽  
Small Ruminants ◽  
Culture Method ◽  
Mycobacterium Avium Subsp ◽  
Type I ◽  
Culture Methods ◽  
Specific Pcr ◽  
Mycobacterium Avium Subsp Paratuberculosis

Paratuberculosis is a chronic disease of ruminants caused by Mycobacterium avium subsp. Paratuberculosis (MAP). Since isolation of MAP type I (S) is rarely reported in Italy, our research was aimed at isolating, by an inexpensive liquid culture manual method, this type of MAP isolates. At first, we used an ELISA to point out to serologically positive samples from five flocks. Secondly, we used a fecal direct IS900-qPCR on the ELISA positive samples, in order to detect shedder animals. Feces from IS900-qPCR positive samples were inoculated in solid and liquid culture media. IS900-qPCR was further used to test the growth of MAP isolates in liquid medium, which were further confirmed by f57-qPCR and submitted to typing by specific PCR in order to identify the MAP type. Twenty-eight samples (24 fecal and four tissutal samples) were processed by culture methods, resulting in the isolation of six type I MAP field isolates. Notably, no isolates were recovered by solid media, underlining the utility of this liquid method. Few data about this type of MAP are currently available in Italy, and further analyses should be carried out in order to study the origin and epidemiology of type I strains circulating in Italy.

Comparison of a Real-Time PCR Method with a Culture Method for the Detection of Salmonella enterica Serotype Enteritidis in Naturally Contaminated Environmental Samples from Integrated Poultry Houses

2012 ◽  
Vol 75 (4) ◽  
pp. 743-747 ◽  
Author(s):  
BWALYA LUNGU ◽  
W. DOUGLAS WALTMAN ◽  
ROY D. BERGHAUS ◽  
CHARLES L. HOFACRE
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Salmonella Enteritidis ◽  
Culture Method ◽  
Pcr Assay ◽  
Culture Methods ◽  
The Real ◽  
Selective Enrichment ◽  
Conventional Culture ◽  
Field Samples

Conventional culture methods have traditionally been considered the “gold standard” for the isolation and identification of foodborne bacterial pathogens. However, culture methods are labor-intensive and time-consuming. A Salmonella enterica serotype Enteritidis–specific real-time PCR assay that recently received interim approval by the National Poultry Improvement Plan for the detection of Salmonella Enteritidis was evaluated against a culture method that had also received interim National Poultry Improvement Plan approval for the analysis of environmental samples from integrated poultry houses. The method was validated with 422 field samples collected by either the boot sock or drag swab method. The samples were cultured by selective enrichment in tetrathionate broth followed by transfer onto a modified semisolid Rappaport-Vassiliadis medium and then plating onto brilliant green with novobiocin and xylose lysine brilliant Tergitol 4 plates. One-milliliter aliquots of the selective enrichment broths from each sample were collected for DNA extraction by the commercial PrepSEQ nucleic acid extraction assay and analysis by the Salmonella Enteritidis–specific real-time PCR assay. The real-time PCR assay detected no significant differences between the boot sock and drag swab samples. In contrast, the culture method detected a significantly higher number of positive samples from boot socks. The diagnostic sensitivity of the real-time PCR assay for the field samples was significantly higher than that of the culture method. The kappa value obtained was 0.46, indicating moderate agreement between the real-time PCR assay and the culture method. In addition, the real-time PCR method had a turnaround time of 2 days compared with 4 to 8 days for the culture method. The higher sensitivity as well as the reduction in time and labor makes this real-time PCR assay an excellent alternative to conventional culture methods for diagnostic purposes, surveillance, and research studies to improve food safety.

scholarly journals Comparing the Yield of Staphylococcus aureus Recovery with Static versus Agitated Broth Incubation

2018 ◽  
Vol 2018 ◽  
pp. 1-3
Author(s):  
Carol E. Muenks ◽  
Patrick G. Hogan ◽  
Carey-Ann D. Burnham ◽  
Stephanie A. Fritz
Keyword(s):  
Staphylococcus Aureus ◽  
Enrichment Culture ◽  
Culture Method ◽  
Ambient Air ◽  
Built Environments ◽  
Culture Methods ◽  
Semiquantitative Assessment ◽  
Direct Plating ◽  
Static Conditions ◽  
Broth Enrichment

Given the lack of standardization of methodologies for microbial recovery from built environments, we sought to compare the yield of Staphylococcus aureus with a broth enrichment method when incubated in agitated versus static conditions. Five unique strains of S. aureus at five different concentrations were cultured to compare direct plating, agitated broth enrichment, and static broth enrichment culture methods. All samples were incubated at 35° in ambient air. The lowest concentration recovered across three replicates and five strains did not differ between culture methods (Fisher’s exact test, p=0.50); notably, recovery of S. aureus was equivalent between static and agitated broth incubation. When broth enrichment was used (both static and agitated), the burden of S. aureus growth was higher (by semiquantitative assessment of 4-quadrant streaking) compared to the direct plating culture method. Optimizing strategies for microbial recovery is essential, particularly in areas of lower biomass, given the paucity of research concerning microbial communities of built environments. The results of this study, in conjunction with other experiments investigating microbiomes of built environments, can help inform protocols for standardizing culturing methods within built environments.

A Real-Time PCR Assay for the Detection of Salmonella in a Wide Variety of Food and Food-Animal Matrices†

2007 ◽  
Vol 70 (5) ◽  
pp. 1080-1087 ◽  
Author(s):  
V. M. BOHAYCHUK ◽  
G. E. GENSLER ◽  
M. E. McFALL ◽  
R. K. KING ◽  
D. G. RENTER
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Culture Method ◽  
Pcr Assay ◽  
Culture Methods ◽  
The Real ◽  
Conventional Culture ◽  
Food Animal ◽  
Highly Sensitive ◽  
Field Samples

Conventional culture methods have traditionally been considered the “gold standards” for the isolation and identification of foodborne pathogens. However, culture methods are labor-intensive and time-consuming. We have developed a real-time PCR assay for the detection of Salmonella in a variety of food and food-animal matrices. The real-time PCR assay incorporates both primers and hybridization probes based on the sequence of the Salmonella invA gene and uses fluorescent resonance energy transfer technology to ensure highly sensitive and specific results. This method correctly classified 51 laboratory isolates of Salmonella and 28 non-Salmonella strains. The method was also validated with a large number of field samples that consisted of porcine feces and cecal contents, pork carcasses, bovine feces and beef carcasses, poultry cecal contents and carcasses, equine feces, animal feeds, and various food products. The samples (3,388) were preenriched in buffered peptone water and then selectively enriched in tetrathionate and Rappaport-Vassiliadis broths. Aliquots of the selective enrichment broths were combined for DNA extraction and analysis by the real-time PCR assay. When compared with the culture method, the diagnostic sensitivity of the PCR assay for the various matrices ranged from 97.1 to 100.0%, and the diagnostic specificity ranged from 91.3 to 100.0%. Kappa values ranged from 0.87 to 1.00, indicating excellent agreement of the real-time PCR assay to the culture method. The reduction in time and labor makes this highly sensitive and specific real-time PCR assay an excellent alternative to conventional culture methods for surveillance and research studies to improve food safety.

scholarly journals Same-Day Simultaneous Diagnosis of Bacterial and Fungal Infections in Clinical Practice by Nanopore Targeted Sequencing

2020 ◽  
Author(s):  
Ming Wang ◽  
Aisi Fu ◽  
Ben Hu ◽  
Gaigai Shen ◽  
Ran Liu ◽  
...  
Keyword(s):  
Clinical Practice ◽  
Bacterial Infections ◽  
Fungal Infections ◽  
Culture Method ◽  
Targeted Sequencing ◽  
Therapeutic Monitoring ◽  
Rrna Gene ◽  
Culture Methods ◽  
Detection Range ◽  
Sequencing Platform

AbstractBACKGROUNDAs approximately 19% of global deaths are attributable to infectious diseases, early diagnosis of infection is very important to reduce mortality. Traditional infection detection strategies have limited sensitivity, detection range, and turnaround times; a detection technology that can simultaneously detect bacterial and fungal infections within 24 h is urgently need in clinical settings.METHODSWe developed nanopore targeted sequencing (NTS) for same-day simultaneous Diagnosis of fungal and bacterial infections. NTS was developed by amplification of 16s rRNA gene (for bacteria), IST1/2 gene (for fungal), and rpoB (for Mycobacterium spp.) using multiple primers, and sequenced by a real-time nanopore sequencing platform. An in-house bioinformatic analyze pipeline was used to diagnose the infectious pathogens by mapping the sequencing results with the constructed databases.RESULTSComparison of 1312 specimens from 1257 patients using NTS and culture method; NTS detected pathogens in 58.71% of specimens from patients, compared to 22.09% detected using the culture method. NTS showed significantly higher sensitivity than culture methods for many pathogens. Importantly, a turnaround time of <24 h for all specimens, and a pre-report within 6 h in emergency cases was possible in clinical practice. Modification of antibiotic therapy and maintenance of original anti-infection regimens in 51.52% (17/33) and 36.36% (12/33) of patients was in accordance with NTS results, and quantitative monitoring of clinical treatment effects was evaluated in four patients by continuous NTS tests.CONCLUSIONSApplication of NTS in clinically detected pathogens can improve targeted antibiotic treatment and therapeutic monitoring.

scholarly journals Profile of Producers and Production of Dry-Aged Beef in Brazil

Foods ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 2447
Author(s):  
Jonatã Henrique Rezende-de-Souza ◽  
Flavio Andre Bolini Cardello ◽  
Ana Paula Moraes de Paula ◽  
Felipe A. Ribeiro ◽  
Chris R. Calkins ◽  
...  
Keyword(s):  
Production Process ◽  
Water Loss ◽  
Relative Frequency ◽  
Product Safety ◽  
Air Velocity ◽  
Aging Period ◽  
Main Indicator ◽  
Chamber Temperature ◽  
Governmental Institutions ◽  
Microbiological Analyses

No information is currently available on the profile of producers and production process of dry-aged beef in Brazil, to the best of the authors’ knowledge. We surveyed 37 Brazilian companies that were producing dry-aged beef in 2020 to investigate this market. The absolute and relative frequency of responses was calculated to obtain the sum, average, minimum, and maximum values. From the respondents, dry-aged beef was first produced in 2009, and most producers are located in big cities. Most respondents control and monitor chamber temperature; however, humidity and air velocity only are monitored. The aging period (mostly between 22 to 60 days) was the main indicator of product readiness. The process losses (water loss and crust trimming) can reach 65%. Some producers perform microbiological analyses to ensure product safety and others use tools such as GMP and SOP. The results of this survey may help governmental institutions to develop a standardized industrial protocol for producing dry-aged beef in Brazil.

Direct detection ofCampylobacter jejuniin human stool samples by real-time PCR

2008 ◽  
Vol 54 (9) ◽  
pp. 742-747 ◽  
Author(s):  
Shiyong Lin ◽  
Xinying Wang ◽  
Haoxuan Zheng ◽  
Zhengguo Mao ◽  
Yong Sun ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Direct Detection ◽  
Culture Method ◽  
Turnaround Time ◽  
Culture Methods ◽  
Time Saving ◽  
Whole Process ◽  
Pure Cultures ◽  
Stool Samples

Our purpose was to establish a quick and accurate real-time PCR (rtPCR) method to detect Campylobacter jejuni directly from human diarrheal stool as an alternative to traditional culture methods. To determine the consistency of rtPCR and culture method, 256 clinical diarrheal stool samples and 50 normal stool samples from healthy individuals were examined, and the whole process was double-blinded. Our data showed that the sensitivity of rtPCR in pure cultures and stool was 102CFU·mL–1and 103CFU·g–1, respectively. Of the 256 diarrheal samples, 10 specimens were successfully detected by both methods, whereas two specimens were PCR positive but culture negative. No positive results were found by these two methods in 50 normal specimens. Our data suggested that rtPCR was convenient in operation and time-saving (turnaround time 3.5–4 h), so it could be used for clinical diagnostic and epidemiological purposes.

scholarly journals Bovine tuberculosis: Diagnosis by PCR technique in bovine milk samples

2013 ◽  
Vol 37 (2) ◽  
pp. 156-160
Author(s):  
Mohammed J. Alwan
Keyword(s):  
Bovine Milk ◽  
Culture Method ◽  
Bacterial Isolation ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Culture Methods ◽  
Milk Samples ◽  
Tuberculosis Diagnosis ◽  
Pcr Method ◽  
Polymerase Chain

     The aim of this study was to determine the percentage of borne tuberculin infection in milk sample by using PCR.  (102) milk samples were collected from  cows , AL-Dejella (39) samples,  AL-Suara (20) samples cow station, AL-Fthalia (20) samples,  AL-Azezia (11) samples and AL-Twarege (12) samples (Iraq) during the period  July 10th   2010 to  Nov.30th   2010. The samples were examined by direct smear stained by Ziehle-Neelson stain, culture methods and confirmed the isolates by Polymerase Chain Reaction (PCR) assay. The results showed that  5, 102 (4.9%) milk samples were M. bovis positive that were detected by direct milk smear method and 10 out 102(9.8%) M.bovis +ve were detected by culture method and PCR assay. The results also showed that high percentage of bacterial isolates from milk samples AL-Dejella city show (12.8%) by culturing and PCR method followed by AL-Suara (10%), AL-Fthelia (10%), Al-Twarege (8.3%) but no bacterial isolation was recorded in AL-Azezia milk samples. This study concluded that M.bovis infection was spreading in dairy cow within the mentioned areas and PCR was more sensitive, rapid, and accurate technique for M.bovis infection diagnosis.

Sign in / Sign up

Export Citation Format

Share Document