scholarly journals Validation of a Modified Version of Actero™ Salmonella Enrichment Media for Rapid Detection of Salmonella spp. in Environmental and Food Samples

2020 ◽  
Vol 103 (5) ◽  
pp. 1326-1337
Author(s):  
Afia Boumail ◽  
Alex Eyraud ◽  
Mounia Akassou ◽  
Mélanie Geffroy ◽  
Jean-Félix Sicard ◽  
...  
Keyword(s):  
Food Samples ◽  
Culture Broth ◽  
Probability Of Detection ◽  
Alternative Methods ◽  
Pet Food ◽  
Pcr Assay ◽  
Salmonella Spp ◽  
Environmental Surfaces ◽  
Chicken Carcass ◽  
Detection Assay

Abstract Background Actero™ Salmonella Enrichment Media1 (Actero™ Salmonella) is a culture broth developed to recover Salmonella spp. from foods and environmental surfaces. Performance of Actero™ Salmonella broth has already been assessed and validated (AOAC Performance Tested MethodSM 041403) for the detection of Salmonella spp. in various foods, feeds and environmental samples. Objective This study aimed to validate the performance of a modified version of Actero™ Salmonella broth by incorporating one of the two liquid supplements into the powdered formula. Methods Inclusivity, exclusivity, stability, and lot-to-lot studies were carried out. Raw ground beef, chicken carcass rinse, dry pet food and stainless steel samples were enriched for 14–20 h at 35–39°C and analyzed using real-time PCR assay as well as by direct plating. Results The Probability of Detection assay confirmed the equivalent performance of the alternative methods as compared to the reference methods. All Salmonella strains, except Salmonella II : 57: z29:-, were able to grow in Actero™ Salmonella broth. One-half of the non-target strains did not grow in Actero™ Salmonella broth, whereas the atypical for Salmonella growth was observed for other non-target microorganisms subsequently plated onto selective and differential agars. Lot-to-lot consistency was demonstrated for three consecutively manufactured lots of the broth. The liquid broth was proven to be stable at 4°C for up to 9 weeks of storage. Conclusions and Highlights The incorporation of one of the two specific supplements into a powdered formula of Actero™ Salmonella broth made it more convenient to use without compromising the performance and accuracy.

Comparative Evaluation of Veriflow® Salmonella Species to USDA and FDA Culture-Based Methods for the Detection of Salmonella spp. in Food and Environmental Samples

2017 ◽  
Vol 100 (5) ◽  
pp. 1445-1457
Author(s):  
Amrita Puri ◽  
Adam C Joelsson ◽  
Shawn P Terkhorn ◽  
Ashley S Brown ◽  
Zara E Gaudioso ◽  
...  
Keyword(s):  
Stainless Steel ◽  
Ceramic Tile ◽  
Ground Beef ◽  
Probability Of Detection ◽  
Salmonella Spp ◽  
Raw Meat ◽  
Environmental Surfaces ◽  
Study Results ◽  
Significant Difference ◽  
The U.S

Abstract Veriflow®Salmonella species (Veriflow SS) is a molecular-based assay for the presumptive detection of Salmonella spp. from environmental surfaces (stainless steel, sealed concrete, plastic, and ceramic tile), dairy (2% milk), raw meat (20%fat ground beef), chicken carcasses, and ready-to-eat (RTE) food (hot dogs). The assay utilizes a PCR detection method coupled with a rapid, visual, flow-based assay that develops in 3 min post-PCR amplification and requires only an 18 h enrichment for maximumsensitivity. The Veriflow SS systemeliminates the need for sample purification, gel electrophoresis, or fluorophore-based detection of target amplification and does not require complex data analysis. This Performance Tested MethodSM validation study demonstrated the ability of the Veriflow SS method to detect low levels of artificially inoculated or naturally occurring Salmonella spp. in eight distinct environmental and food matrixes. Ineach reference comparison study, probability of detection analysis indicated that there was no significant difference between the Veriflow SS method and the U.S. Department of Agriculture Food Safety and Inspection Service MicrobiologyLaboratory Guidebook Chapter 4.06 and the U.S. Food and Drug Administration Bacteriological Analytical Manual Chapter 5 reference methods. A total of 104 Salmonella strains were detected in the inclusivity study, and 35 nonspecific organisms went undetected in the exclusivity study. The study results show that the Veriflow SS method is a sensitive, selective, and robust assay for the presumptive detectionof Salmonella spp. sampled from environmental surfaces (stainless steel, sealed concrete, plastic, and ceramic tile), dairy (2% milk), raw meat (20% fat ground beef), chicken carcasses, and RTE food (hot dogs).

Evaluation of the 3M™ Molecular Detection Assay (MDA) 2 – Listeria monocytogenes for the Detection of Listeria monocytogenes in a Variety of Foods and Select Environmental Surfaces: Collaborative Study, First Action 2016.08

2017 ◽  
Vol 100 (2) ◽  
pp. 454-469
Author(s):  
Patrick Bird ◽  
Jonathan Flannery ◽  
Erin Crowley ◽  
James Agin ◽  
David Goins ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Confidence Interval ◽  
Molecular Detection ◽  
Reference Method ◽  
Probability Of Detection ◽  
Chicken Breast ◽  
Inoculum Level ◽  
Test Portion ◽  
Environmental Surfaces ◽  
Detection Assay

Abstract The 3M™ Molecular Detection Assay (MDA) 2 – Listeria monocytogenes uses loop-mediated isothermal amplification of unique DNA target sequences combined with bioluminescence to rapidly detect Listeria monocytogenes in a broad range of food types and on environmental surfaces. Using an unpaired study design, technicians from 13 laboratories located in the United States and Canada compared the 3M MDA 2 – Listeria monocytogenes to the U.S. Department of Agriculture Food Safety Inspection Service Microbiology Laboratory Guidebook Chapter 8.09 “Isolation and Identification of Listeria monocytogenes from Red Meat, Poultry, and Egg Products, and Environmental Samples” reference method for the detection of L. monocytogenes in deli turkey and raw chicken breast fillet. Each matrix was evaluated at three levels of contamination: an uninoculated control level (0 CFU/test portion), a low inoculum level (0.2–2 CFU/test portion), and a high inoculum level (2–5 CFU/test portion). Statistical analysis was conducted according to the probability of detection (POD) statistical model. Results obtained for the low inoculum level test portions produced a difference in the collaborating laboratory POD (dLPOD) value of 0.04 with a 95% confidence interval of (−0.08, 0.17) for deli turkey, indicating that the difference between methods was not statistically significant at the 0.05 probability level. For raw chicken breast fillet, a dLPOD value of 0.16 with a 95% confidence interval of (0.04, 0.28) indicated a statistically significant difference, with an observed higher proportion of positive results by the candidate method compared to the reference method.

Roka Listeria Detection Method Using Transcription Mediated Amplification to Detect Listeria Species in Select Foods and Surfaces

2012 ◽  
Vol 95 (6) ◽  
pp. 1672-1688 ◽  
Author(s):  
Hua Yang ◽  
Shannon Kaplan ◽  
Michael Reshatoff ◽  
Ernie Hu ◽  
Alexis Zukowski ◽  
...  
Keyword(s):  
Probability Of Detection ◽  
Culture Methods ◽  
Department Of Agriculture ◽  
Detection Analysis ◽  
Reference Methods ◽  
Smoked Salmon ◽  
Environmental Surfaces ◽  
Detection Assay ◽  
Inspection Service ◽  
The U.S

Abstract The Roka Listeria Detection Assay was compared to the reference culture methods for nine select foods and three select surfaces. The Roka method used Half-Fraser Broth for enrichment at 35 ± 2°C for 24–28 h. Comparison of Roka's method to reference methods requires an unpaired approach. Each method had a total of 545 samples inoculated with a Listeria strain. Each food and surface was inoculated with a different strain of Listeria at two different levels per method. For the dairy products (Brie cheese, whole milk, and ice cream), our method was compared to AOAC Official MethodSM993.12. For the ready-to-eat meats (deli chicken, cured ham, chicken salad, and hot dogs) and environmental surfaces (sealed concrete, stainless steel, and plastic), these samples were compared to the U.S. Department of Agriculture/Food Safety and Inspection Service-Microbiology Laboratory Guidebook (USDA/FSIS-MLG) method MLG 8.07. Cold-smoked salmon and romaine lettuce were compared to the U.S. Food and Drug Administration/Bacteriological Analytical Manual, Chapter 10 (FDA/BAM) method. Roka's method had 358 positives out of 545 total inoculated samples compared to 332 positive for the reference methods. Overall the probability of detection analysis of the results showed better or equivalent performance compared to the reference methods.

Method Modification for the Atlas Listeria Environmental LE Detection Assay Using FoodChek Actero Listeria Enrichment Media and Half-Fraser Media for the Detection of Listeria spp. from Environmental Surfaces

2018 ◽  
Vol 101 (2) ◽  
pp. 562-576
Author(s):  
Brett Maroni ◽  
Tucker Lopez ◽  
Cambria Neal ◽  
Sarah Verver ◽  
Celina Puente ◽  
...  
Keyword(s):  
Stainless Steel ◽  
Reference Method ◽  
Common Species ◽  
Sample Volume ◽  
Probability Of Detection ◽  
Department Of Agriculture ◽  
Environmental Surfaces ◽  
Detection Assay ◽  
Inspection Service ◽  
High Level

Abstract Two candidate method modifications for the Atlas Listeria Environmental LE Detection Assay were compared with the U.S. Department of Agriculture (USDA)-Food Safety and Inspection Service Microbiology Laboratory Guidebook 8.09 (MLG 8.09) method for detection of Listeria spp. on stainless steel, polyvinyl chloride (PVC), and sealed concrete surfaces. For LE candidate method 1, samples were enriched in FoodChek Actero Listeria Enrichment Media [ALEM; Performance Tested MethodSM (PTM) 111201] at 35 ± 2°C for 18 to 24 h and evaluated for a range of analytical sample volumes. For LE candidate method 2, the current Roka PTM using 90 mL of Half-Fraser broth for enrichment at 35 ± 2°C was evaluated at 24 h with a reduced sample volume. These comparisons were made in multiple studies across the three environmental surfaces. Within each method and study, a total of 5 samples were uninoculated, 20 samples were inoculated with Listeria spp. at a low level to target fractional positivity, and 5 samples were inoculated with Listeria spp. at a high level to approach a probability of detection of 1. Inclusivity and exclusivity studies were also conducted for the LE method in combination with Half-Fraser and ALEM. The Atlas Listeria Environmental LE Detection Assay detected all 50 inclusive organisms, including 25 strains of L. monocytogenes and 5 strains of each of the other five common species of Listeria (L. innocua, L. welshimeri, L. ivanovii, L. seeligeri, and L. grayi) and none of the 30 exclusive organisms across all media and with both 200 and 2000 µL sample volumes. For the LE candidate method 1 studies, no significant differences were observed within the Roka ALEM method at 18, 20, or 24 h and for both the 200 and 2000 µL sample volumes as compared with the paired culture outcome. However, the ALEM method performed significantly better as compared with the unpaired reference method for sealed concrete and stainless steel. For the LE candidate method 2 studies, no significant differences were observed within the Roka HF method at 24 h for the 200 and 2000 µL samples as compared with the paired culture outcomes and unpaired reference method outcomes across the surfaces. The independent laboratory studies observed no significant differences in performance between the USDA/MLG 8.09 reference method and candidate methods 1 or 2, respectively, across the evaluated parameters. Overall, the candidate method 1 modification parameters and candidate method 2 sample parameters for the Atlas Listeria Environmental LE Detection Assay were statistically equivalent to or better than the reference method for detection of Listeria spp. on stainless steel, PVC, and sealed concrete surfaces, providing greater flexibility in method application for end users.

Evaluation of 3M Molecular Detection Assay (MDA) 2–Listeria for the Detection of Listeria Species in Select Foods and Environmental Surfaces: Collaborative Study, First Action 2016.07

2017 ◽  
Vol 100 (1) ◽  
pp. 82-98
Author(s):  
Patrick Bird ◽  
Jonathan Flannery ◽  
Erin Crowley ◽  
James Agin ◽  
David Goins ◽  
...  
Keyword(s):  
Molecular Detection ◽  
Collaborative Study ◽  
Reference Method ◽  
Probability Of Detection ◽  
Chicken Breast ◽  
Test Portion ◽  
Listeria Species ◽  
Environmental Surfaces ◽  
Detection Assay ◽  
Significant Difference

Abstract 3M Molecular Detection Assay (MDA) 2–Listeria uses loop-mediated isothermal amplification and bioluminescence detection to rapidly detect Listeria species in a broad range of food types and environmental surfaces. Using an unpaired study design, MDA 2–Listeria was compared with the U.S. Department of Agriculture, Food Safety and Inspection Service's Microbiology Laboratory Guidebook Chapter 8.09 “Isolation and identification of Listeria monocytogenes from red meat, poultry and egg products, and environmental samples” reference method for the detection of Listeria in deli turkey and raw chicken breast fillet. Technicians from 13 laboratories located within the continental United States and Canada participated in the collaborative study. Each matrix was evaluated at three levels of contamination: uninoculated control (0 CFU/test portion), low inoculum (0.2–2 CFU/test portion), and high inoculum (2–5 CFU/test portion). Statistical analysis was conducted according to the probability of detection (POD) statistical model. Results obtained for the low-inoculum-level test portions produced a difference between two laboratory POD values (dLPOD) with 95% confidence intervals of 0.04 (–0.08, 0.17) for deli turkey, indicating the difference between the methods was not statistically significant at the P = 0.05. For raw chicken breast fillet, a dLPOD value with 95% confidence interval of 0.16 (0.04, 0.28) indicated a statistically significant difference between the two methods, with an observed higher proportion of positive results by the candidate method than the reference method.

Evaluation of the MicroSEQ™ Salmonella spp. Detection Kit with the PrepSEQ™ Rapid Spin Sample Preparation Kit for Detection of Salmonella spp. in Dry Pet Food

2016 ◽  
Vol 99 (2) ◽  
pp. 401-406 ◽  
Author(s):  
Jonathan Cloke ◽  
Jonathan Flannery ◽  
Benjamin Bastin ◽  
Patrick Bird ◽  
Erin Sutphin Crowley ◽  
...  
Keyword(s):  
Sample Preparation ◽  
Reference Method ◽  
Statistical Test ◽  
Probability Of Detection ◽  
Contamination Level ◽  
Pet Food ◽  
Salmonella Spp ◽  
Test Portion ◽  
High Level ◽  
Assay Protocol

Abstract A method modification validation study was conducted to validate the Applied Biosystems MicroSEQ™ Salmonella spp. Detection Kit for the detection of Salmonella spp. in 375 g samples of dried pet food. The MicroSEQ assay protocol, using the Applied Biosystems PrepSEQ™ Rapid Spin DNA Sample Preparation Kit, was compared to the reference method detailed in the U.S Food and Drug Administration (FDA) Bacteriological Analytical Manual (BAM; Chapter 5, Salmonella) for detection of Salmonella spp. For each method, 20 replicates were analyzed at a low contamination level of 0.2–2 CFU/test portion, five replicates were analyzed at a high level of contamination of 2–5 CFU/test portion, and five control replicates were also analyzed at 0 CFU/test portion (uninoculated). Statistical analysis was conducted using the Probability of Detection statistical test to determine the ability of the MicroSEQ Salmonella spp. Detection Kit to detect Salmonella from 375 g samples of dried pet food in comparison to the FDA-BAM reference method. The results demonstrated that the MicroSEQ Salmonella spp. Detection Kit was able to accurately detect Salmonella spp. present in dry pet food after an enrichment time of 20 h.

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.

Multiplex PCR Detection of Enterotoxin Genes in Aeromonas spp. from Suspect Food Samples in Northern Taiwan

2008 ◽  
Vol 71 (10) ◽  
pp. 2094-2099 ◽  
Author(s):  
YU-CHANG CHANG ◽  
JAN-YI WANG ◽  
AMMAIYAPPAN SELVAM ◽  
SHU-CHEN KAO ◽  
SHANG-SHYNG YANG ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Diarrheal Disease ◽  
Simultaneous Detection ◽  
Food Poisoning ◽  
Food Samples ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Enterotoxin Genes ◽  
Causative Agents ◽  
Northern Taiwan

Aeromonads possess an array of virulence factors and are causative agents of a number of human infections. Among them, genes of one cytotoxic (Act) and two cytotonic (Alt, Ast) enterotoxins are implicated in a human diarrheal disease. A rapid, specific, simultaneous detection of these enterotoxin genes in suspected food poisoning samples is not yet reported. Hence, a multiplex PCR assay was designed to amplify the cytotoxic (act), heat-labile cytotonic (alt), and heat-stable cytotonic (ast) enterotoxin genes of aeromonads. The PCR assay was tested with 133 Aeromonas spp. isolated from suspect food poisoning samples and retail samples of poultry and fish from wet markets in and around Taipei, Northern Taiwan. The Aeromonas spp. isolates were divided into six genotypes based on absence or presence of one or more enterotoxin genes. Of these 133 isolates, Aeromonas caviae (52.5%) and Aeromonas hydrophila (43.4%) were the most frequently isolated species from food poisoning samples and retail samples, respectively. Among the species, A. hydrophila had a significantly higher proportion for harboring three enterotoxin genes than had the others, whereas Aeromonas encheleia, considered a nonpathogen, was found harboring three enterotoxin genes. The multiplex PCR assays are rapid and specific, and provide a useful tool for the detection and genotyping of enterotoxin genes of aeromonads.

scholarly journals Occurrence, antimicrobial susceptibility patterns and genotypic relatedness of Salmonella spp. isolates from captive wildlife, their caretakers, feed and water in India

2018 ◽  
Vol 146 (12) ◽  
pp. 1543-1549
Author(s):  
Arockiasamy Arun Prince Milton ◽  
Rajesh Kumar Agarwal ◽  
Govindarajan Bhuvana Priya ◽  
Cheruplackal Karunakaran Athira ◽  
Mani Saminathan ◽  
...  
Keyword(s):  
Antimicrobial Susceptibility ◽  
Animal Species ◽  
Multidrug Resistant ◽  
Wild Animal ◽  
Diffusion Method ◽  
Pcr Assay ◽  
Direct Pcr ◽  
Salmonella Spp ◽  
Culture Methods ◽  
Faecal Samples

AbstractOccurrence of Salmonella spp. in captive wild animal species in India is largely unknown. The purpose of this study was to determine the occurrence of different Salmonella serotypes, antimicrobial resistance patterns and genotypic relatedness of recovered isolates. A total of 370 samples including faecal (n = 314), feed and water (n = 26) and caretakers stool swabs (n = 30) were collected from 40 different wild animal species in captivity, their caretakers, feed and water in four zoological gardens and wildlife enclosures in India. Salmonellae were isolated using conventional culture methods and tested for antimicrobial susceptibility with the Kirby–Bauer disc diffusion method. Salmonella isolates were serotyped and genotyping was performed using enterobacterial repetitive intergenic consensus (ERIC) PCR and 16S rRNA sequencing. Animal faecal samples were also subjected to direct PCR assay. Salmonella was detected in 10 of 314 (3.1%) faecal samples by isolation and 18 of 314 (5.7%) samples by direct PCR assay; one of 26 (3.8%) feed and water samples and five of 30 (16.7%) caretakers stool swabs by isolation. Salmonella was more commonly isolated in faecal samples from golden pheasants (25%; 2/8) and leopard (10%; 2/20). Salmonella enterica serotypes of known public health significance including S. Typhimurium (37.5%; 6/14), S. Kentucky (28.5%; 4/14) and S. Enteritidis (14.3%; 2/14) were identified. While the majority of the Salmonella isolates were pan-susceptible to the commonly used antibiotics. Seven (43.7%; 7/16) of the isolates were resistant to at least one antibiotic and one isolate each among them exhibited penta and tetra multidrug-resistant types. Three S. Kentucky serotype were identified in a same golden pheasants cage, two from the birds and one from the feed. This serotype was also isolated from its caretaker. Similarly, one isolate each of S. Typhimurium were recovered from ostrich and its caretaker. These isolates were found to be clonally related suggesting that wildlife may serve as reservoir for infections to humans and vice versa. These results emphasise the transmission of Salmonella among hosts via environmental contamination of feces to workers, visitors and other wildlife.

scholarly journals Occurrence and Antimicrobial Susceptibility Profile of Salmonella Isolates from Animal Origin Food Items in Selected Areas of Arsi Zone, Southeastern Ethiopia, 2018/19

2021 ◽  
Vol 2021 ◽  
pp. 1-13
Author(s):  
Minda Asfaw Geresu ◽  
Behailu Assefa Wayuo ◽  
Gezahegne Mamo Kassa
Keyword(s):  
Antimicrobial Susceptibility ◽  
Control Strategies ◽  
Raw Milk ◽  
Food Samples ◽  
Animal Origin ◽  
Salmonella Spp ◽  
Food Items ◽  
Antimicrobial Susceptibility Profile ◽  
Arsi Zone ◽  
Animal Origin Food

The status of Salmonella and its antimicrobial susceptibility profile in animal origin food items from different catering establishments in Ethiopia is scarce. Hence, this study aimed to investigate the occurrence and antimicrobial susceptibility profile of Salmonella isolates from animal origin food items in the selected areas of Arsi Zone. One hundred ninety-two animal origin food samples were collected and processed for Salmonella isolation. Isolates were tested for their susceptibility to 13 antimicrobials using Kirby–Bauer disk diffusion assay. An overall prevalence of 9.4% (18/192) Salmonella spp. isolates were recovered from animal origin food samples collected from different catering establishments. Seven (21.9%) of “Dulet,” 4 (12.5%) of “Kitfo,” 3 (9.4%) of “Kurt,” 2 (6.3%) of raw milk, 1 (3.1%) of egg sandwich and 1 (3.1%) of cream cake samples were positive for Salmonella. Catering establishments, protective clothing, source of contamination, manner of hand washing, and money handling were among the putative risk factors that were significantly associated ( p < 0.05 ) with Salmonella spp. occurrence. Ampicillin, nitrofurans, and sulphonamide resistance were significantly associated ( p < 0.05 ) with Salmonella spp. occurrence in the selected food items. Three (16.7%), 5 (27.8%), 5 (27.8%), and 4 (22.2%) of the isolates were resistant to 3, 4, 5, and 6 antibiotics, respectively, whereas only a sole isolate was resistant to two antibiotics (viz. ampicillin and kanamycin). In conclusion, the general sanitary condition of the catering establishments, utensils used, and personnel hygienic practices were not to the recommended standards in the current study. Besides, detection of multidrug-resistant strains of Salmonella in animal origin food items from different catering establishments suggests the need for detailed epidemiological and molecular characterization of the pathogen so as to establish the sources of acquisition of resistant Salmonella strains. Hence, implementation of Salmonella prevention and control strategies from farm production to consumption of animal origin food items are crucial.

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