METHOD COMPARISON STUDY

Transcription

1 METHOD COMPARISON STUDY Detection of Salmonella spp. in Dry Pet Food and Environmental Matrices using Elite Salmonella Enrichment Media and Real-Time PCR Assay for Salmonella Prepared by Sergiy Olishevskyy, PhD Vice President, Research and Development FoodChek Laboratories Inc. 7755, av Duplessis Saint-Hyacinthe, Quebec J2R 1S5 Canada Phone: This method s performance was reviewed by AOAC RESEARCH INSTITUTE and was found to perform to the manufacturer s specifications.

2 Dry pet foods and treats are at risk for contamination by various pathogens, especially Salmonella, because they often contain raw materials derived from animal origins and each ingredient may be processed under different conditions. Protect Your Pets and Family From Infection About 80 million households in the United States own a dog or a cat, representing 65% of the US households. Many pet owners feed their companion animals with dry pet food because of the ease of use and nutritional benefit. Dry pet foods and treats often contain raw materials derived from animal origin and each ingredient may be processed under different conditions. These items are at risk for contamination by various pathogens, especially Salmonella. Reduce the Risk of Salmonella Contamination in Dry Pet Food Contaminated dry pet food that is not handled properly can also cause serious illness in people, especially children. Regardless, pet owners should always wash their hands after handling dry food and treats to manage the risk of contamination. 2 The Only Solution for Detecting Salmonella in Dry Pet Food Due to sub-lethally injured Salmonella cells surviving the processing treatment of dry pet foods, an optimized method of recovery and enrichment of the bacterium is needed. The Actero Elite Salmonella Enrichment Media provides for effective recovery of injured an/or dormant Salmonella cells from different lowmoisture food samples after a single-step enrichment. Advantages of Advantages of Actero Actero Elite Salmonella Elite Salmonella Enrichment Enrichment Media Media Single enrichment reduces time-to-results and lab costs High level of accuracy and reliability Results in an amazing 18 hours of enrichment time Release your products in less than 24 hours Uses one-third less enrichment media per volume compared to standard enrichment media Detect using Real-Time PCR Assay for Salmonella AOAC validated for 375 g composite sample sizes, the new industry-standard test portion size for dry pet food

3 Elite Salmonella Enrichment Media and Real-Time PCR Assay for Salmonella is the ONLY solution for You Results in an amazing 18 hours of enrichment time Achieve real cost reduction by converting to Actero TM Elite Salmonella Enrichment Media Protect your brand, your customers, their families, and pets Actero Elite Salmonella Enrichment Media is the proven solution for problem matrices! FoodChek s Actero Elite Salmonella Enrichment Media has received AOAC PTM CERTIFICATION for use with Real-Time PCR Assay for Salmonella 3

4 Technical Report Table of Contents The Problem with Salmonella in Dry Pet Food 5 Method Comparison Study Report, Easy & Rapid Detection of Salmonella in Dry Pet Food 6 Abstract 6 Materials 6 Method 7 Results 8 Conclusions 8 Accelerate Your Real-Time PCR Assay for Salmonella Results with Actero Elite Salmonella Enrichment Media 9 It s Easy to Convert to Actero Elite Salmonella Enrichment Media 10 Actero Elite Salmonella Enrichment competitive advantages. 11 Actero Elite Enrichment Media product list. 12 Appendix 13 Actero Elite Enrichment Media compatible with any diagnostic system. 14 Actero Elite Enrichment Media and Real-Time PCR Assays for Listeria and Salmonella. 15 4

5 THE PROBLEM WITH SALMONELLA IN DRY PET FOOD There is growing interest among pet owners to feed their companion animals processed dry pet food because of the ease of use and its nutritional benefits. As a result, the consumer market has put increasing pressure on many manufacturers to raise production volumes and expand their product assortment while also ensuring the safety for consumption and handling. Dry pet foods and treats are often made with animal-origin products that could be a host for Salmonella or other organisms. In general, these products are cooked to temperatures that will kill these organisms. However, if a contaminated additive (a flavoring, for example) is added to the food after cooking, or if the food comes in contact with contaminated materials, the food will consequently become contaminated. An emerging concern with respect to Salmonella carriage in pets is the role of natural pet treats and raw food diets, which are produced with little to no regulatory oversight in the United States and Canada. There is a lack of information on how natural pet treats and raw food diets can affect the health of pets and their owners, both in general and with respect to Salmonella contamination. Over the past number of years, there have been reports of dry pet food contaminated with Salmonella that caused sickness in pets and also humans, including infants with yet to be fully developed immune systems, who handled the food and treats. As a result, in 2013 the U.S. FDA adopted zero tolerance for Salmonella in dry pet food. This enforcement has pushed the dry pet food industry to routinely test for Salmonella in dry pet food and treats. For all these reasons, FoodChek has developed Actero Elite Salmonella Enrichment Media to selectively enrich Salmonella in dry pet foods and treats for its further detection. FoodChek understands the challenging regulatory and business demands on dry pet food producers along with the importance of volume testing and inventory release that protects your bottom line, without compromising product safety or the integrity of your brand. The method to enrich and detect Salmonella using Actero Elite Salmonella Enrichment Media was developed with input from the dry pet food industry to offer the best practical solution for a monitoring and surveillance program for Salmonella. Despite the inhibitory challenges associated with this sample matrix, Actero Elite Salmonella Enrichment Media has been certified by AOAC Research Institute Performance Tested Method, License Number Actero Elite Salmonella Enrichment Media is an approved dry pet food and enviromental testing solution that is designed to accurately screen for Salmonella with exceptional specificity and speed, helping to reduce costly false positives and inventory holds. With fast, reliable, and easily implemented testing workflows, FoodChek can help you: Minimize your risk Maximize your competitive edge Reduce your production costs Protect your brand, your customers, their families, and pets 5

6 METHOD COMPARISON STUDY REPORT EASY & RAPID DETECTION OF SALMONELLA IN DRY PET FOOD Abstract FoodChek Systems Inc. developed an effective method for easy and rapid detection of Salmonella in dry pet food samples using Actero Elite Salmonella Enrichment Media with Real-Time PCR Assay for Salmonella, which has successfully been granted AOAC PTM CERTIFICATION. The method protocol was developed and validated for two sizes of dry pet food samples including 25 g and 375 g (usually used for composite protocol). The results of the study demonstrated that performance of the candidate method is statistically equivalent to the reference culture method for detecting Salmonella in dry pet food. The time to detect Salmonella spp. has been reduced up to 20 hrs. including the enrichment and detection steps when the Actero Elite Salmonella Enrichment Media was used. No false negative and false positive results were observed. The high accuracy and reliability of the method were confirmed in the independent laboratory validation study. Materials Actero Elite Salmonella Enrichment Media Dehydrated format: FCM g bottle and three supplement vials Ready to use liquid formats: FCM L MediaBox FCM L MediaBox (with supplements added) API 20 E kits for identification of enterobacteriacae Brilliant Green dye 1% solution Real-Time PCR Assay for Salmonella Hektoen Enteric (HE) agar Lactose broth Rappaport-Vassiliadis (RV) broth Salmonella serogroup E antiserum Tetrathionate broth (TT) Trypticase Soy agar plates with 5% of sheep blood agar Triton X-100 Xylose Lysine Deoxycholate (XLD) agar Phosphate Buffered Saline (PBS) 6

7 METHOD COMPARISON STUDY REPORT EASY & RAPID DETECTION OF SALMONELLA IN DRY PET FOOD Method 7

8 METHOD COMPARISON STUDY REPORT EASY & RAPID DETECTION OF SALMONELLA IN DRY PET FOOD Results Probability of detection (POD) statistical model was used to evaluate the differences between presumptive and confirmed results as well as between the alternative and the reference methods. The results of the method comparison validation study for both 25 and 375 g samples of dry pet food are summarized in the table below. POD analysis didn t show any significant differences (the 95% confidence interval of the dpods contains zero) in the method performance between Real-Time PCR Assay for Salmonella and the reference method. No false positive or false negative results were observed. Matrix CFU/Test Portion N Real-Time PCR Assay for Salmonella Method US FDA BAM Chapter 5 Method Confirmed POD C Confirmed POD R dpod CR Dry Pet Food (25 g) Dry Pet Food (375 g) Conclusions The results of the study demonstrated that performance of Real-Time PCR Assay for Salmonella in dry pet food samples enriched with Actero Elite Salmonella Enrichment Media is equivalent to the reference method. It has been proven that Actero Elite Salmonella Enrichment Media is able to selectively enrich Salmonella for its further detection in the shortest possible time. 8

9 Environmental Samples Food Samples ACCELERATE YOUR REAL-TIME PCR ASSAY FOR SALMONELLA RESULTS WITH ACTERO ELITE SALMONELLA ENRICHMENT MEDIA Enrichment of Salmonella spp. in Actero Elite Salmonella Media vs other media prior to Real-Time PCR Assay for Salmonella. Matrix Actero TM Elite Salmonella Method Media A Method Type Size Time Medium volume Time Medium volume Dry pet food 25g h 225 ml Not validated Dry pet food 375 g h 2625 ml h 3375 ml Stainless steel 100 cm h 90 ml h 225 ml Plastic 100 cm h 90 ml h 225 ml Actero Elite Salmonella Enrichment Media Highlights TM Actero Elite Salmonella can reduce sample enrichment time by approximately 20% Achieve real cost reduction by converting to Actero Elite Salmonella Enrichment Media Enrichment time is 14 to 30 hrs depending the matrix Actero Elite Salmonella Enrichment Media is a standalone media system and compatible with the Real-Time PCR Assay for Salmonella This method s performance was reviewed by AOAC RESEARCH INSTITUTE and was found to perform to the manufacturer s specifications. 9

10 IT S EASY TO CONVERT TO ACTERO ELITE SALMONELLA ENRICHMENT MEDIA The FoodChek Team specializes in food microbiology that helps labs and processors to achieve optimum efficiency in controlling pathogens throughout the food processing chain. The dedicated team of FoodChek professionals have the expertise in facilitating the adoption of Actero Elite Salmonella Enrichment Media in your plant by providing you support in every step of the conversion process. As your pet food safety partner, the FoodChek team assists you in implementing the best possible approach for the control and the monitoring of Salmonella in your dry pet food production. FoodChek understands the importance of achieving rapid time-to-results without compromising accuracy and safety. The FoodChek Team is well positioned to provide customized service solutions designed for your specific requirements. Protect your brand, your customers, their families, and pets. 10

11 Elite Salmonella Enrichment Media competitive advantages. Single-step enrichment Fastest time-to-results Accurate Reduce sample handling Use less enrichment media* Improve workflow Higher throughput Release food products sooner * Depends on matrix 11

12 Salmonella United States Canada Europe Elite Salmonella Enrichment Media product list. Our patented Actero Elite Salmonella Enrichment Media is a selective bacterial culture medium specifically optimized for the recovery of Salmonella spp. from environmental and food samples in a single-step enrichment. We continue to strive to be the food industry s single, trusted provider for pathogen and other microorganism testing supplies and equipment. Actero Elite Salmonella Enrichment Media is compatible with a RT-PCR Assay for Salmonella and represents the result of true food safety innovation. Microorganism FoodChek Cat. # Product Name Format Description FCM-009 ACTERO ELITE SALMONELLA ENRICHMENT MEDIA 500G 500g dehydrated media and 3 supplement vials (yields 30L rehydrated media). FCM-010 ACTERO ELITE SALMONELLA/STEC ENRICHMENT MEDIA 500G A selective bacterial culture medium specifically optimized for recovery of Salmonella spp. and Shiga-toxin producing Escherichia coli (STEC) from environmental and food samples in a single-step enrichment. 500g dehydrated media and 3 supplement vials (yields 30L rehydrated media). FCM-047 ACTERO ELITE SALMONELLA MEDIABOX 5L A selective bacterial culture medium specifically optimized for recovery of Salmonella spp. from environmental and food samples in a single-step enrichment in liquid format. FCM-048 ACTERO ELITE SALMONELLA MEDIABOX 10L A selective bacterial culture medium specifically optimized for recovery of Salmonella spp. from environmental and food samples in a single-step enrichment in liquid format. For a free sample, contact us at: Global: North America Toll Free: info@foodcheksystems.com 12

13 APPENDIX 13

14 Elite Enrichment Media compatible with any diagnostic system. Current Listeria spp., Listeria monocytogenes and Salmonella spp. Validations Diagnostic Instrument External Validation License/ Certification # Pathogen Matrix Method Developer AOAC PTM (3) Listeria spp. and Listeria monocytogenes Variety of food and environmental surfaces BAX System RT-PCR (1) AOAC PTM (3) Salmonella spp. Health Canada (5) MFLP-79 Listeria spp. Variety of food and environmental matrices Stainless steel, ceramic, plastic, sealed concrete, and rubber Hygiena, LLC Atlas System (2) AOAC PTM (3) Listeria spp. Stainless steel, PVC and sealed concrete Roka Bioscience, Inc. Agilent AriaMx AFNOR (4) EGS 38/ AOAC PTM (3) , & Listeria spp. and Listeria monocytogenes Listeria spp. and Listeria monocytogenes Variety of food and environmental surfaces Variety of food and environmental surfaces Eurofins GeneScan Bio-Rad CFX96 Touch DeepWell AFNOR (4) EGS 38/ AOAC PTM (3) , & Listeria spp. and Listeria monocytogenes Listeria spp. and Listeria monocytogenes Variety of food and environmental surfaces Variety of food and environmental surfaces Eurofins GeneScan Xpress LM AOAC PTM (3) Listeria monocytogenes Stainless steel, plastic, ceramic tile Crystal Diagnostics MICT Reader Health Canada (5) MFLP-99 Listeria spp. Environmental surfaces AOAC PTM (3) Listeria spp. Environmental surfaces FoodChek Systems Inc. AOAC PTM (3) Salmonella spp. Variety of egg formats and environmental surfaces (1) Formerly an asset of DuPont Nutrition & Health (2) Acquired by IEH Laboratories & Consulting Group (3) This method s performance was reviewed by AOAC RESEARCH INSTITUTE and was found to perform to the manufacturer s specifications. (4) This method s performance was reviewed by AFNOR and was found to perform to the manufacturer s specifications in accordance with the NF Validation general rules and NF102 certification rules. (5) Method has been approved for publication in the HEALTH CANADA Compendium of Analytical Methods. 14

15 Elite Enrichment Media and Real-Time PCR Assays for Listeria and Salmonella validated matrices. ACTERO ELITE LISTERIA ENRICHMENT MEDIA ACTERO ELITE SALMONELLA ENRICHMENT MEDIA AOAC RI-PTM License # AOAC RI-PTM License # (SUITE) AOAC RI-PTM License # AOAC RI-PTM License # (SUITE) Internally Validated Concrete/Sealed Fish/Raw 25 g Stainless Steel Almonds/Raw 375 g Flour/Wheat 25 g Stainless Steel Ham/Cured 125 g Plastic Beef/Ground 25 g Plastic Ice Cream 25 g Cheese/Soft Mexican Style Frankfurters Salmon/Cold Smoked Shrimp/Frozen Cooked Spinach/Fresh Bagged 25 g Milk/Pasteurized 25 g 125 g Turkey/Smoked Breast Chocolate/ Liquor Chocolate/Milk Bar 125 g Cocoa/Powder 25 g 25 g Eggs/Shell 25 g Beef/Ground 375 g 25 g Butter/Peanut 25 g 20 Eggs Chicken/Carcass Rinse Chicken/Ground 30 ml 25 g Pet Food/Dry 25 g Egg/Dried Whole 100 g 25 g Pet Food/Dry 375 g Egg/Liquid Whole* 25 g 100 g Onions/ Dehydrated Pepper/Black Whole 25 g 375 g Parsley/Dried Pepper/Black Whole 25 g 25 g Raisins/Dried 25 g *Pasteurized 15

16 Elite Salmonella Enrichment Media and Real Time-PCR Assay for Salmonella cutting edge food safety testing solutions. FoodChek Systems Inc. Global: North America Toll Free: Website: foodcheksystems.com United States Carl (Chip) Zerr III Vice President, Product Integration Phone: