Solus One Salmonella AOAC extension to scope Herbs

Solus One Salmonella AOAC extension to scope – Herbs, spices and flavorings

“ One game changing element is the single step media enrichment process which has saved us hours of analysis time while also delivering high quality results to our customers so they can put their products into the market with confidence. ” Erin Crowley Chief Scientific Officer at Q Laboratories

Solus One Salmonella - AOAC Extension to scope One enrichment • Simple protocols • Rapid Results One enrichment • Reduced media requirement when compared to BAM • Solus proprietary m. BPW • Solus supplement (2. 22 ml/L)

Different dilutions… Paprika Powder Whole black peppercorns Cinnamon Powder Honey mustard seasoning Flavored ranch seasoning 25 g 375 g 1 in 10 1 in 50 1 in 10 225 ml 1225 ml 3375 ml BAM methods Paprika/Black pepper – 1 in 10 TSB, 1 hour incubation at RT followed by 22 -26 hours at 35°C Cinnamon – 1 in 100 TSB, 1 hour incubation at RT followed by 22 -26 hours at 35°C

… same method Simple protocols • • • 20 -24 hours (upto 3 hrs faster than BAM) 41. 5± 1°C (selective for Salmonella) Same proprietary enrichment media for all certified herb/spice matrices Rapid Results • • Post-enrichment time = 2 hours TTR = <24 hours including assay time

Important aspects of the extension protocols The herb/spices extension protocols use proprietary Solus modified BPW m. BPW supplemented at 2. 22 ml/L (half volume required for other matrices) Herbs/spices already have antimicrobial effect therefore less supplement is required to inhibit non-target growth (keeps costs down) m. BPW provides better growth support to Salmonella and Solus supplement provides inhibition to Gram positive flora to achieve appropriate detection levels

Solus One Salmonella Supplement Proprietary selective supplement Allows growth of stressed/injured Salmonella Suppresses the growth of majority of Gram positive flora Contains a novel selective component* A significant improvement over using selective dyes e. g. brilliant green Allows isolation of Salmonella from primary enrichment cultures Supplied freeze dried * Currently subject to patent application.

Updated AOAC kit inserts • Updated to include instructions for herb/spice matrices • 5 plate kit – Insert 30 (on Q pulse) • 1 plate kit – Insert 31 (on Q pulse) AFNOR kit inserts • remain as inserts 21 and 28 (5 plate and 1 plate, respectively) without herb/spice details.

AOAC certified methods for herb/spice testing Test Name AOAC Certified Spices/Herbs Solus One Salmonella 25 g whole black peppercorns, 25 g paprika powder, 25 g cinnamon, 375 g honey mustard seasoning, 375 g flavored ranch seasoning 3 M Molecular Detection Assay 2 - Salmonella 25 g black pepper ACTERO Salmonella/STEC Enrichment Media 25 g dried parsley, 25 g whole black pepper ANSR for Salmonella 25 g Black pepper Atlas Salmonella Detection Assay 25 g black pepper, 25 g nacho cheese seasoning BAX System PCR Assay for Salmonella 25 g black pepper foodproof Salmonella Detection Kit (liquid & lyphilized) foodproof Short. Prep I kit, Star. Prep One OR Magnetic Preparation Kit I with Robo. Prep series White pepper, cumin Gene. Disc Plate Salmonella spp. 25 g black pepper Gene. Quence Salmonella Microwell Assay Black pepper GENE-UP Salmonella 25 g and 375 g garlic powder Micro. SEQ Salmonella spp. Detection Kit 25 g black pepper Pathatrix Salmonella species pooling (with culture detection) 25 g black ground pepper Singlepath Salmonella Lateral Flow Assay 25 g black pepper Thermo Scientific Sure. Tect Salmonella 25 g black peppercorns Transia Plate Salmonella Gold 25 g ground black pepper Veriflow Salmonella species 25 g ground black pepper VIDAS® Salmonella (SLM) Assay Method Easy SLM with Chrom. ID™ Salmonella (SM 2) Agar 25 g ground black pepper VIDAS UP 25 g ground black pepper

Possible customer questions What does an extension to scope mean? Solus One Salmonella already has AOAC PTM certification for select food and environmental matrices. Solus submitted further test results to AOAC for new matrices (paprika powder, whole black peppercorns, cinnamon, honey mustard onion seasoning and flavored ranch seasoning) to determine that Solus One Salmonella also detects Salmonella in these matrices with the sensitivity and selectivity required. These results were accepted by AOAC and these matrices are now included in the certification remit. Do I need to re-validate the whole assay? No, there is no need to re-validate the whole assay. If the customer is already using the assay and wishes to start testing herbs/spices with Solus One Salmonella then they will need to perform a small validation procedure for the required matrices only.

Possible customer questions Can I use BPW rather than m. BPW? No. The methods for testing herbs/spices are slightly different due to the challenging nature of pathogen testing in these matrices. The modified BPW is required to better support the recovery and growth of injured Salmonella which may be present in these matrices. Standard BPW may not support the pathogen cells sufficiently to detect their presence in a sample, leading to false negative results. Why is the Solus supplement volume different for herbs/spices than for other matrices? The natural antimicrobial properties of herbs/spices provides a certain level of selectivity in these matrices, therefore a lower volume of supplementation is required to suppress the growth of Gram positive flora.

Possible customer questions Has anything in the kit changed? The contents of the Solus One Salmonella kit have not been altered as part of this extension to scope. Which Salmonella species does Solus One Salmonella detect in herbs and spices? During testing for the extension the following Salmonella species and subspecies were tested: Salmonella species Salmonella enterica Salmonella bongori Salmonella subspecies Salmonella enterica subspecies enterica Salmonella enterica subspecies salamae Salmonella enterica subspecies arizonae Salmonella enterica subspecies diarizonae Salmonella enterica subspecies houtenae Salmonella enterica subspecies indica

S O L U S S C I E N T I F I C K I T M A T R I C E S

Summary • AOAC method updated to include paprika, pepper, cinnamon, honey mustard seasoning and flavored ranch seasoning • Reduced media requirement compared to BAM • Incubation up to 3 hours shorter than BAM • 2. 22 ml/L supplement in m. BPW for all herb/spice matrices • 41. 5°C incubation temperature for all matrices • New kit inserts awaiting internal approval on Q pulse