Attachment_24.pdf

Uncontrolled when printed on: 10/30/2014 For the most current and official copy, check the Master List.

FIELD BULLETIN #30 – FOOD PROGRAM AREA

INSTRUCTIONS FOR ENVIRONMENTAL SAMPLING

OCTOBER 2014

Subject: Instructions for Environmental Sampling Equipment;

Identification of Equipment Sources; Sample Collection Technique for Large and Small Area Environmental Surface Sampling and District Response to Positive Samples

Effective Date: Immediately.

Supersedes all other versions of DDFI Field Bulletin 30

Target Audience: DIBs, SCSOs and all CSOs performing food program, assignments

Purpose: Beginning in FY 09, there has been an increased emphasis on conducting bacteriological inspections in the food arena, as certain foods pose an increased risk of association with Listeria monocytogenes and Salmonellae. Since its implementation in FY 10, environmental sampling has been a valuable tool in enhancing food safety.

This bulletin was developed to standardize the procedures for sampling the environment and equipment, which may be supplemented with finished product sample collection.

OFFO prepared this bulletin with the assistance of SANLAB, CFSAN, and ORA/ORS.

Specific instructions for when to collect environmental samples will be contained in the work plan, compliance program or assignment instructions. If a CSO is unsure of when or under what circumstances to perform environmental sampling, they should consult with their supervisor.

CFSAN has developed guidance on the specific locations within a processing facility to collect environmental samples to increase the likelihood of detecting contamination with Listeria monocytogenes and Salmonellae.

Since the original issuance of this Bulletin, the agency remains concerned about the presence of Listeria monocytogenes, and Salmonellae in food processing plant environments as a result of positive environmental samples and foodborne illness associated with these pathogens, likely due to contamination from the environment. FDA is not currently aware of situations in which pathogenic Esherichia coli (such as E. coli O157:H7 or other Shiga-toxin producing E. coli have colonized the manufacturing environment and led to contamination of food. However, there may be situations in which targeted environmental sampling may be recommended for investigation purposes (see page 6 for additional information).

Background/Recommendations

For environmental sampling, the broth or buffer serves two purposes, 1) to neutralize sanitizer that may be on surfaces that you are sampling, and 2) to provide nutritional requirements for the organisms of interest to recover from stress and injury and to survive transit to the laboratory.

http://inside.fda.gov:9003/downloads/ProgramsInitiatives/FieldOperations/FieldGuidance/ucm022967.pdf http://inside.fda.gov:9003/downloads/ProgramsInitiatives/FieldOperations/FieldGuidance/ucm022966.pdf

D/E (Dey and Engley) neutralizing broth has been shown to be effective as a neutralizing agent against the widest range of sanitizing agents that may be in use by a firm, and per CFSAN and ORA/ORS, is the one to be used for general purpose environmental sampling. Attachment 1 of this bulletin is a memo prepared by Dr. Donald Zink with further detail on the choices made regarding sampling equipment.

If there are questions on the use of D/E broth/buffer vs. other broths or buffers; or any other questions on the sampling reagents or methodology please contact:

Email ORA/ORS at ORAHQ ORS MANAGEMENT@fda.hhs.gov; Gary Hartman, Microbiologist, SANLAB, at 510-337-6745 or Gary.Hartman@fda.hhs.gov; or Dr. Donald Zink, Senior Science Advisor, Office of the Center Director, CFSAN, Office of Food Safety at 240-402-1693 or Donald.Zink@fda.hhs.gov.

When collecting environmental samples you should make every effort to collect as much sample material as possible. A sample will consist of multiple subsamples consisting of sponge and/or swab samples of specific sample sites and will be referred to as “subs” unless describing sponge or swab samples specifically. For Salmonellae environmental sampling collect at least 100 subs and ideally 300 subs if there are a sufficient number of potential sample sites. For Listeria environmental swabbing, collect at least 50 swabs and ideally 100 or more subs if there are a sufficient number of potential sample sites. Salmonellae tend to be more difficult to detect in a contaminated facility vs. Listeria and a greater number of samples are needed for Salmonellae environmental sampling in order to have confidence in negative findings.

Environmental samples are collected from the processing plant environment in order to fully evaluate the environment and detect even low levels of contamination. It is not unusual for a contaminated plant to yield only 1 – 2% positive environmental subs. Most environmental samples should be collected using either hand held sponges or sponges on a stick. Larger samples or larger areas sampled are more likely to be positive. Swabs are suitable for sampling only very small areas that cannot be accessed any other way.

Sample Equipment Sources

Sources for sponges, sponge-sticks, and swabs pre-hydrated with D/E neutralizing buffer or broth can be found in Attachment 2. Specific products are identified, but equivalent products can be used such as, unhydrated sponges and sticks and single use tubes of D/E neutralizing broth.

ORA/ORS is recommending the use of 70% ethyl alcohol as the preferred disinfectant for gloves and equipment. See Attachment 2 for supplier sources.

Other general sampling supplies you will need for environmental sampling may include:

• Sterile gloves for sample collector

• Hand sanitizers – 70% ethyl alcohol (wash and sanitize hands often during sampling)

• 70% Filtered USP Ethyl Alcohol to sanitize gloves between subs (note change from isopropyl alcohol previously recommended)

• Cooling medium for samples/frozen gel packs mailto:ORAHQORSMANAGEMENT@FDA.HHS.GOV mailto:Gary.Hartman@fda.hhs.gov mailto:Donald.Zink@fda.hhs.gov

• Coolers or insulated boxes

• Labels to identify samples

• Permanent marker

• Flashlight

• Sterile metal spatulas (small) or other sterile implement to scrape debris out of cracks

• Clean paper towels

• Whirlpack or equivalent single-use sterile bags

• Overnight shipping labels

• New trash bags to line cooler, set supplies on in facility, and for discarded wrappers

• Measuring device, ruler, tape measure

For Zone 1 sampling in Kosher facilities where using a milk protein base is prohibited, use sterile phosphate buffer solution or sterile saline solution. See Attachment 2 for supplier sources.

Selection of Areas to Collect Environmental Samples

During the initial phases of the inspection, the CSO should conduct a walk-through assessment observing and mapping operations, including the location of equipment, flow of the product, foot traffic of employees, forklift/mule traffic patterns, and segregation of raw material versus finished products, and consider sampling areas where food is exposed and being processed, particularly post-treatment/pasteurization. Determine the processing zones, as described below. The “Zone Concept” identifies and prioritizes processing areas from highest risk and closest to the product to lowest risk and farthest from the product for potential contamination and harboring growth and niches for targeted pathogen and therefore should be implemented upon conducting environmental sampling as follows:

• Zone 1: Zone 1 refers to all direct food contact surfaces such as slicers, mixers, conveyors, utensils, racks, work tables, etc. When it has been determined that Salmonellae is the pathogen of concern, food contact surfaces in Zone 1 recently cleaned or that look clean are normally not sampled unless specifically requested in the assignment or CPG. In contrast, when Listeria monocytogenes is the determined as the pathogen of concern, sampling of food contact surfaces in Zone 1 is a must.

• Zone 2: Zone 2 encompasses the areas directly adjacent to food contact surfaces (Zone 1). For investigations focusing on Salmonellae, this is the area where environmental contamination is most likely to lead to contamination of Zone 1 and affect the product. In a small production room, Zone 2 encompasses all non-food contact surfaces in the processing area, such as the exterior of equipment, framework, food carts, equipment housing, gears, ventilation and air handling equipment, and floors. In a much larger room (e.g. 20,000 square feet) Zone 2 is the area around the exposed product in which you could envision a pathway to product contamination either through the actions of man or machine;

for example, even a far corner of the room could be considered Zone 2 if foot traffic or forklifts move through that area and these traffic patterns also go very near a line where exposed food is conveyed or held, or ventilation patterns cause airflow from these remote areas. For Salmonellae sampling, the most productive sample sites tend to be on the floor, or very near the floor on equipment surfaces, and potential harborage areas. Thus, potential harborage areas in low-lying sites should be given the greatest emphasis. However, because food processing plants differ greatly in their design, construction, and manufacturing processes, CSOs should exercise discretion and feel free to sample any site that may have collected airborne Salmonellae or that could be a harborage/growth site for Salmonellae or Listeria.

• Zone 3: Zone 3 is the area immediately surrounding Zone 2. Zone 3 is an area that, if contaminated with a pathogen, could lead to contamination of zone 2 via actions of humans or movement of machinery. Examples of Zone 3 areas include corridors and doorways leading into food production areas or areas in a large production room that are further away from food handling equipment than typical Zone 2 areas. Walls, phones, forklifts and “mules”, even if physically located in Zone 2, should be considered Zone 3 due to a decreased likelihood of product contamination.

• Zone 4: The area immediately surrounding Zone 3, generally considered a remote area. Zone 4 is an area which, if contaminated with a pathogen, could lead to contamination of Zone 3 via the actions of humans or machinery.

Examples of Zone 4 areas include an employee locker room if not immediately adjacent to food production rooms, dry goods storage warehouse, finished product warehouse, cafeterias, hallways, and loading dock area.

For Listeria, a large majority of the environmental samples collected should be taken from Zones 1 and 2, and to a much lesser degree, Zone 3 areas. Generally, Zone 4 areas are unproductive for Listeria and positive findings in Zone 4 are very difficult to link to a risk of product contamination; thus no, environmental samples should be taken from Zone 4 areas.

For Salmonellae, all environmental samples should be taken from Zones 2 and 3.

Generally, Zone 1 areas are unproductive for Salmonellae because these are not areas where the organism grows and finds harborage. Zone 1 surfaces are usually cleaned too frequently (or “flushed” with product) to become a harborage for Salmonellae. Zone 4 areas can be productive sites for the detection of Salmonellae; however, it can be difficult to link positive findings in zone 4 areas to a direct risk of product contamination.

For this reason, it is recommended that no environmental samples be collected from Zone 4.

Document the possible link between the environmental sample and possible contamination of the food product (supporting the Food, Drug and Cosmetic Act Section 402(a)(4)). Describe the location of the sample in relation to areas where food is exposed and being processed, particularly post-treatment/pasteurization and any mechanical or human activities you observe that might cause an organism to be spread.

Document and illustrate your observations and the locations of environmental samples using diagrams, photographs, and journal notes.

Other Considerations

When walking through the plant, as in all inspections, be mindful of the possibility of transporting pathogens into the facility, and from Zones 3 and 4 into Zone 2 or Zone 1.

The firm should rely on cleaning and sanitization procedures and restricting movement of employees and equipment from Zones 3 and 4 to other areas of the facility to mitigate the transfer of Salmonellae or other pathogens from this zone to adjacent areas. The adequacy of controls in place to address this should be determined as part of the GMP inspection during collection of environmental samples, and discussed with management if the firm has not implemented these controls.

Follow the firm’s procedures with respect to using disposable shoe coverings and using foot disinfection baths or door foamers prior to entering these areas. To minimize the potential for cross contamination and to focus sample collection initially on the most relevant zones, it is recommended that sampling start at Zone 1 (for Listeria monocytogenes) and Zone 2 for Salmonellae. When passing over food processing equipment on catwalks, stairs, etc., it is especially critical that disposable shoe coverings are worn so as not to contaminate food contact surfaces with moisture, dirt, etc. from your shoes. Lab jackets or Tyvek suits, and shoes, must be clean when beginning the inspection and especially when entering Zone 2 from other areas. To that end, work shoes must be cleaned and sanitized prior to leaving the office or home and carried in a clean container to the work site. Work shoes (leather) should be cleaned with leather cleaner and the soles should be washed with soap and water and then sanitized with a chlorine solution or use an equivalent sanitizer such as quaternary ammonium following the labeled instructions on the container. Change into the work shoes inside the facility.

See IOM Chapter 5 sections for Clothing and General Inspection Procedures.

Where to Target Sampling for Listeria monocytogenes

Listeriosis is a foodborne illness of major public health concern because of the severity of the disease (meningitis, septicemia, and pregnancy complications such as miscarriage or stillbirth), a high case-fatality rate, and a long incubation period. Listeria monocytogenes differs from most other food-borne pathogens because it is widely distributed, resistant to diverse environmental conditions, including low pH and high NaCl concentrations, and is microaerobic and psychrotrophic. The various ways it can enter food processing plants; its ability to grow and survive for long periods of time in the environment, on foods, and in food processing plants; and its ability to grow at low temperatures and to survive in or on food for prolonged periods under adverse conditions, have made it a major concern for many manufacturing industries in recent decades.

Perform most of the sampling for Listeria in, on, and around food contact equipment;

focusing on areas where food is exposed and being processed, particularly post-treatment/pasteurization. Every effort should be made to conduct sampling when the facility has been in production for at least four hours and before any wet cleaning is performed.

See Attachment 3 for guidance on the specific locations within a processing facility to collect environmental samples to increase the likelihood of detecting contamination with Listeria monocytogenes.

References:

1. Doyle, Michael et al. Food Microbiology: Fundamentals and Frontiers 2nd Ed.

Pgs 383-403.

2. Cliver, Dean and Riemann, Hans. Foodborne Diseases 2nd Ed. Pgs 55 – 67.

3. FDA Bad Bug Book 2nd Ed. Listeria monocytogenes http://www.fda.gov/Food/FoodborneIllnessContaminants/CausesOfIllnessBadBu gBook/ucm2006773.htm

4. Guidance for Industry: Control of Listeria monocytogenes in Refrigerated or Frozen Ready-To-Eat Foods; Draft Guidance. February 2008.

http://www.fda.gov/food/guidanceregulation/guidancedocumentsregulatoryinform ation/foodprocessinghaccp/ucm073110.htm

Where to Target Sampling for Salmonellae

Salmonellae survive drying particularly well and are difficult to kill with heat when they are dehydrated or in low moisture products such as chocolate and peanut butter. Look for Salmonellae in areas of the production facility that occasionally get wet, but that dry out for long periods of time before any wet cleaning is performed. Specific sampling procedures for large and small areas are described later in this Bulletin. Proper sampling for Salmonellae in a food plant involves spending a great deal of time on your hands and knees – the low lying areas of the plant are where the organism is most often found.

See Attachment 4 for guidance the specific locations within a processing facility to collect environmental samples to increase the likelihood of detecting contamination with Salmonellae.

References:

1. Doyle, Michael et al. Food Microbiology: Fundamentals and Frontiers 2nd Ed. Pgs 141-178.

2. Cliver, Dean and Riemann, Hans. Foodborne Diseases 2nd Ed. Pgs 55 – 67.

Environmental Sample Collection for E. coli O157:H7

At this time, CFSAN and OFFO are not including instructions on collecting environmental samples for E. coli O157:H7. As background, there have been investigations associated with outbreaks of E. coli O157:H7 where environmental sampling has been conducted.

To date, the organism has not been isolated from these samples. It is current thinking that it does not readily colonize processing environments, but is most likely a contaminated ingredient or product issue and may be present as a transient pathogen.

Currently, there are no validated methods for analyzing environmental samples for E.

coli O157:H7. For this reason, it is requested that no environmental samples be collected for analysis for this pathogen on a surveillance basis. Should there be a for-cause basis identified, please consult with OFFO or CFSAN program contacts to obtain concurrence and instructions. Any decision to conduct environmental sampling for E. coli O157:H7 will be made on a case-by-case basis by the CORE foodborne disease investigation team in consultation with CFSAN/ORA HQ subject matter experts.

Generally, specific instructions will be provided through an assignment should the decision be made to proceed with sampling.

http://www.fda.gov/Food/FoodborneIllnessContaminants/CausesOfIllnessBadBugBook/ucm2006773.htm http://www.fda.gov/Food/FoodborneIllnessContaminants/CausesOfIllnessBadBugBook/ucm2006773.htm http://www.fda.gov/food/guidanceregulation/guidancedocumentsregulatoryinformation/foodprocessinghaccp/ucm073110.htm http://www.fda.gov/food/guidanceregulation/guidancedocumentsregulatoryinformation/foodprocessinghaccp/ucm073110.htm

Sampling Method

On the first day of the inspection you will need to conduct a walk-through of the entire facility to understand the process flow, the work flow of the employees, and how equipment moves in and out of the Zones.

As a general rule, to minimize the possibility of contaminating the processing area with pathogens, it is better to start in Zones 1 and 2 and work your way out to Zones 3 and 4.

Note: Use aseptic technique throughout the sampling procedure. Be sure to submit closed (unopened) controls, including gloves, sponges, media and swabs in accordance with IOM Chapter 4 section Controls. Follow standard procedures for sampling and evidence development when collecting samples, including information such as, date, size of area sampled, location (diagrams/charts) etc. (See IOM Chapter 4 – Sampling).

Gloves

The policy regarding changing of gloves between each sub during environmental sample collection as depicted in the FD148 Environmental Sampling in Food Manufacturing training video has been revised. The current policy does not require the sample collector to change gloves between each sub in Zones 2 - 4 as long as the sample collector remains in the same zone and the integrity of the gloves is not compromised during the course of collecting the sub, (i.e. glove rips, or if it is brushed against a lab coat, etc.). For example, if 50 subs are collected in Zone 2, the sample collector would not need to change gloves between each of these subs, but would change gloves when moving to another zone, another distinct processing room or area, or if the condition of the gloves warrants changing. However, gloves should be sanitized between each sub by applying a 70% solution of ethyl alcohol. It is expected that collection of a large number of subs in one area would necessitate several changes of gloves. Note: ORS has changed the recommended alcohol sanitizer from isopropyl to ethyl alcohol as they have determined it is more effective. If unavailable at the time of sampling, 70% isopropyl alcohol is still effective and can be used.

For subs collected in Zone 1 continue to follow the established policy and change gloves between each sub as described in the Environmental Sampling training video. Note: If collecting finished product, change gloves between product samples (i.e. for 10 cantaloupes, change gloves between each cantaloupe).

Closed Controls

Submit one unopened package of pre-sterilized equipment (i.e. gloves, D/E broth, whirlpack or equivalent sterile bag) per distinct lot per inspection. It is recommended that this be submitted on the last day of sample collection.

Open Controls

As discussed in the Environmental Sampling training video, do not collect open controls during environmental sampling.

Note: Open controls are still required when collecting in-line samples.

Sample and Sub Numbering

It may take multiple days to collect 100 to 300 subs. Use one sample number per day and number all environmental subs sequentially from 1 to X, where X = the total number of subs. If multiple days are necessary, use a new sample number for each day but continue to number the subs sequentially, i.e. Day 1, Sample 111111, Subs 1 – 150;

Day 2, Sample 111112, Subs 151 – 300. For controls, use alphabetic identification, i.e.

Sample 111111, Subs a, b, c, d, e. Closed controls need only be collected once per facility unless the lot changes, even if samples are collected over multiple days. Submit these to the laboratory on the last day of collection under the first sample number.

Instructions for the Collection Report

Environmental samples have a specific product code. DO NOT assign the commodity code for environmental samples as these are being tracked through FACTS and also by CFSAN, and will be missed. On the C/R, please be sure to indicate the following fields:

Surveillance: Sample collected on an objective basis where there is no inspectional or other evidence of a problem with the product.

Compliance: Sample collected on a selective basis as the result of an inspection, complaint or other evidence that there may be a problem with the product.

Several new product codes have been created to allow for the tracking of environmental samples. See Table 1-Product Codes for Specific Commodities and Field Alert #39 for more information.

Table 1-Product Codes for Specific Commodities

Commodity

Product Code

Foods/Feeds

• 52Y[][]07(Farm Environmental Swabs/Samples)

• 52Y[][]08 (Process/Manufacturing Environmental Swabs/Samples)

• 52Y[][]**(Animal Carcass Rinse/Swabs) o Where **= 01 (Beef), 02 (Chicken), 03 (Lamb), 04 (Pork), 05 Turkey), 06 (Other)

Large area Environmental Surface Sampling Procedure

Large areas are approximately 1 by 1 foot for unclean surfaces; or 3 by 3 feet for cleaned and/or sanitized surfaces. Use this procedure also for equivalent environmental surfaces such as floors, walls, tables, and carts, using a sponge on a stick.

Sampling of dry surfaces with an unhydrated sponge-stick (for an unhydrated sponge, follow instructions, below as appropriate)

Using a felt-tip black permanent marker, label the sterile bag containing the sponge with appropriate sample information (or affix pre-made labels to the outside of the bag).

1. Wash and sanitize your hands to the mid-forearm. Use clean disposable paper towels for drying your hands.

2. From the outside of the sponge bag manipulate the handle toward one side. Pull off the top of the bag holding the Sponge-stick along the perforation. Using the tabs on both sides of the wired band, pull gently to open the sample bag. Do not remove the Sponge-stick.

3. Pour into the Sponge-stick bag 9-10 ml or sufficient volume of DE neutralizing broth on the side away from the handle to hydrate the sponge (do not get broth on the handle). Be careful not to touch the opening of the broth container to any non-sterile surface before or during this transfer.

4. Massage the sponge through the outside of the bag to facilitate absorption.

From the outside of the bag, push the Sponge-stick to the upper portion of the bag. While pushing the sponge-stick up from the bottom of the bag, squeeze excess DE broth from the sponge back into the bag. The sponge should be moist but not dripping wet.

5. Using aseptic technique, unwrap and place a sterile glove upon the hand you will use for swabbing. Do not touch any non-sterile surface (i.e., clothes, skin, counter tops, etc.) with the outside surface of the sterile glove. The other hand can be left ungloved for manipulation of non-sterile surfaces and materials.

6. Remove the sponge-stick from the bag using your gloved hand. Using even and firm pressure push the sponge in one direction across the desired area of the environmental surface 10 times vertically, then 10 times horizontally. If visible soil or residue is present, sample the surface by vigorously rubbing the sponge over the designated area until the soil or residue is removed. Sampling of large flat surfaces (i.e. floor, table tops, and conveyor belts) should cover areas as referenced above, depending on whether the area is unclean or has been cleaned and sanitized. It may be necessary to wet the sponge with additional neutralizing broth when sampling large and/or porous areas. Try to use only enough broth to keep the sponge gliding smoothly over the surface. If there is excess broth squeeze it back into the sample bag and continue until you have sampled the entire sampling site.

7. After sampling, return the sponge to original bag with any excess broth, snap off the handle in accordance with the product instructions that accompany the sponge stick and discard the handle, and submit sponge as a subsample.

8. Follow the policy on use of gloves earlier in this bulletin.

9. Squeeze as much air out of the bag as possible. Roll the top of the bag over several times until it is folded all the way down to the sponge. Fold in the tabs to lock the fold in place. Place the sponge bag inside another empty whirlpack or equivalent bag and seal as before. Both bags must be tight enough to provide both a leak proof seal and minimal airspace during shipment of the moistened sponge.

10. As soon as possible, place the double-bagged sponge inside an insulated cooler, with pre-frozen gel packs to keep the samples cold, but not frozen, and transport/ship the sample to the servicing lab for analysis so it is received by the lab within 24 hours of collection. Do not ship on Friday unless a prior arrangement with servicing laboratory has been made.

Sampling of dry surfaces with a hydrated sponge

Use a felt-tip black permanent marker to label the sterile bag containing the sponge with appropriate sample information.

1. Wash and sanitize your hands to the mid-forearm. Use clean disposable paper towels for drying your hands.

2. Open the sample bag.

3. Place a sterile glove on your hand, as in #5 above.

4. With your sterile gloved hand, remove the sponge from the bag.

5. Swab the sampling surface as in #6 above.

6. Return the sponge to the bag, and proceed as in #8-10 above.

Sampling of Wet Surfaces

Sample using aseptic techniques with a dry sponge or Sponge-stick following the general instructions above, returning the sponge or sponge-stick to the sample bag and using aseptic techniques add 10 ml of DE neutralizing broth to the bag. Proceed as instructed in # 8-10, above.

Small Area Environmental Surface Sampling Procedure

Small areas are less than 1 x 1 foot.

Swabs are suitable for sampling only very small areas that cannot be accessed any other way. For example, the swab can be used to sample the material in a hole in the floor such as might be encountered when a piece of floor mounted equipment is removed from an area and the floor has not been repaired to fill the bolt holes. Swabs may also be useful for sampling floor cracks or the inside of tubular equipment mounts.

Sampling of Dry Surfaces

Collect samples using aseptic techniques with the swab prehydrated with D/E neutralizing broth. Using even and firm pressure, swab in one direction across the desired surface 10 times vertically, then 10 times horizontally, then 10 times diagonally.

If visible soil or residue is present, sample the surface by vigorously rubbing the swab over the designated area until the soil or residue is removed. Return the swab to its swab tube, close tube, place in a whirlpack or equivalent sterile bag, and as soon as possible place inside an insulated cooler with pre-frozen gel pack for transport/shipment to the laboratory within 24 hours of collection. Do not ship on Friday unless a prior arrangement with servicing laboratory has been made.

If you do not have prehydrated swabs, but have swab tubes and vials of D/E neutralizing broth you can use this method, using aseptic technique:

1. Loosen screw cap on D/E neutralizing broth vial (if it is not a flip top container) and the swab tube.

2. Remove sterile swab from the swab tube.

3. Open D/E neutralizing broth vial and pour contents into the swab tube.

4. Insert swab into broth to moisten tip.

5. Collect sample.

6. Place swab into swab tube, close, place into whirlpack or equivalent sterile bag, and submit sample to lab as noted above.

Sampling of Wet Surfaces

Collect sample using aseptic technique using the dry swab in the same manner as noted above. After swabbing, using aseptic technique add D/E neutralizing broth to the swab and transport to laboratory as noted above.

Sampling Debris on Equipment and from Floor Defects, Joints and Gaps

Debris can be sampled by scraping it out using a sterile implement, such as a small metal spatula. Accumulations of processing residues are ideal environments for pathogens such as Salmonellae. Submit debris in D/E neutralizing broth in swab tube following instructions in #1-6 for dry surfaces, but do not use the swab. For larger amounts, a whirlpack or equivalent sterile bag may be used.

Environmental Sampling Video Training

A video demonstrating sample techniques is available on the FDA Intranet at:

http://intranetapps.fda.gov/media/2009/Sampling-11-05-09-508.wmv

Special Instructions for Sampling in Kosher Establishments or Other Facilities Where Milk Proteins May Not be Used

Kosher processors may prohibit the use of a sampling medium such as D/E neutralizing broth that contains a milk protein when swabbing Zone 1 surfaces for Listeria. The following method may be used as a substitute:

1. Moisten a dry sponge (available from several sources) with sterile water, sterile saline or sterile phosphate buffer (any of these are acceptable).

2. Swab the surface.

3. Immediately put the sponge into the whirlpack or equivalent sterile bag and add two 10 ml tubes (total of 20 ml) of D/E neutralizing broth and repeatedly squeeze the sponge to equilibrate the liquid in the sponge.

This procedure will not have a significant negative impact on recovery of the organism as the entire sponge and volume of broth is cultured in the laboratory. In this situation the D/E broth will be diluted (about 66% strength) but it will still be effective.

District Response to Laboratory Findings from Environmental Samples

These procedures are a decision-making framework to guide District and Headquarters staff to implement a consistent national approach to respond to environmental sample results in an effort to streamline corrective actions and public health protection. It is http://intranetapps.fda.gov/media/2009/Sampling-11-05-09-508.wmv important to note that while this broad approach should be applied across all cases, the specific regulatory response taken will be based on a case-by-case review by Headquarters and District staff based on the firm's history, the inspectional findings, the location of the sample, the potential for a route of contamination of final product, the public health risk, and the firm's response to the findings as criteria. For this reason, it is essential that the possible link between the environmental sample and contamination of the food product be documented and clearly presented.

Step 1: Laboratory reports swab(s) as Cannot Rule Out (CRO)

A CRO result is preliminary. CROs may be determined to be negative upon completion.

The time between reporting a sample as CRO and the confirmation of a positive finding is usually 1 to 3 days. Serotyping and the PFGE result require an additional 3 – 5 days from the confirmation report.

Response by the District:

Due to the possibility of CROs reports being confirmed as negative results, do not inform the firm (or the State) of CRO results, which may cause unnecessary reaction by the firm. At the CRO stage, the district should review all documentation related to the CRO environmental sample in preparation for discussion with Headquarters regarding implications for the firm and the products in the market. Documentation review should include floor plans, the process flow, the location of the CRO sub in relation to processing steps and/or proximity to lots of regulated product/exterior of finished product packaging/finished product pallets (this is particularly important for Salmonellae results since the environmental sampling strategy is typically to NOT sample food contact surfaces) and identification of lot numbers for any stored product adjacent to these sub locations. In addition, document whether there are potential routes of cross contamination with finished product, and support with any photographs taken, and 483 items observed, including poor sanitation, employee practices, building construction, and any other relevant evidence.

Step 2: Confirmed Positive Results

When positive or negative environmental samples are reported, the firm is to be notified within 48 hours, and preferably within 24 hours, and advised of the findings.

It is important to report negative findings as well, as some firms may hold product pending results. In all cases, document the notification to the firm and details of the discussion and the firm’s response. Implications of the positive findings should be discussed with firm and include the significance of the location, potential for contamination of product, and the firm’s response to this. The significance of positive findings is dependent on the pathogen identified, and the zone in which the sub was collected, and whether or not a route of contamination of the finished product can be demonstrated. When a conference call (see next steps, below) is necessary, this should be held prior to notification of the firm and the State and should be scheduled in a timely manner to allow the firm’s notification within 48 hours.

Regulatory action under section 402(a)(4) would have to be based on sufficient evidence (such as routes of cross-contamination) to show that the food was prepared, packed, or held under insanitary conditions whereby it may have become contaminated with filth, or whereby it may have been rendered injurious to health.

The next steps will be determined by the assessment of the environmental sample locations:

1. For Listeria monocytogenes in Zone 1 and Salmonellae in Zone 2, a conference call including the District, OFFO, CFSAN OC, CFSAN science advisors, and ORA OE should be scheduled by the District to determine next steps, including whether the firm should be asked to recall the product (OFFO can assist in this if needed). However, a finding in Zone 2 for Listeria monocytogenes and Zone 3 for Salmonellae should have a demonstrated route of contamination to warrant this type of discussion; in all other cases the District should handle and advise OFFO of the outcome. The District should also contact the State regulatory agency and advise of these results and invite the State to participate in the conference call (FDA Commissioned Officers only). See Response by the State below for procedures.

2. In addition, for subs found positive in the above zones, finished product should be collected from the lots processed at that time or at the time of the return visit. Note: when the initial inspection is at a location remote from the RP or District Office, collection of finished product should be considered during the initial inspection, rather than waiting for laboratory results, in order to conserve resources. Follow-up steps should be determined on a case-by-case basis following normal regulatory procedures. The State agency should be given the opportunity to participate in a joint inspection when follow-up inspections are conducted.

3. Following the above discussions, the District Office will contact the firm with the findings and discuss the firm’s next steps and elicit the firm’s response.

For this reason, the conference call must be scheduled within 48 hours and preferably 24 hours so that the firm can be contacted within that timeframe as well.

4. For Listeria monocytogenes in Zones 2 and 3 and Salmonellae in Zone 3, the District should discuss with the firm what acceptable remedial steps would be expected. A re-inspection, which includes collection of environmental samples, should be scheduled within six months to verify the effectiveness of the firm’s clean-up procedures. A strategy call as described in #1 is not needed, but OFFO should be advised of the outcome including a brief description of the findings and the firm’s response.

5. Finished product samples should be collected for all For Cause inspections irrespective of environmental sample results. Note: For finished product samples collected during environmental sampling investigations, do not use the environmental sampling code for product samples on the C/R; the commodity code should be used for these samples.

Response by the State

If a State identifies a confirmed positive environmental sample during a contract inspection/environmental sampling, the State will notify the District State Program coordinator with the results. In the event that the positive environmental sample was taken from Zone 1 (wet environments) or Zone 2 (dry environments) a conference call between the State, District, OFFO, ORA ORS, and the appropriate Center should be held immediately to determine what regulatory actions should be pursued. If the positive sample was not from these zones, a conference call may not be necessary and the

State and District should independently assess the significance of the positive sample, inform the firm, and solicit their response. The State and District would subsequently be charged with communicating the firm’s response and any proposed regulatory action and/or follow-up to ORA HQ and the Center. The State and District should take into account any finished product samples (along with the positive environmental sample) when making the decision to convene a conference call (See Step 2 above).

Also the discussions between the District and State should include possible State regulatory actions either separately or in conjunction with FDA actions (see below), and if the State or District should do a joint follow-up visit or inspection to verify the corrective actions, or a single agency follow-up.

The District and the State should discuss environmental sample positives done under the State inspection/sampling program regarding notification of the District, follow-ups, regulatory actions etc. to ensure continued collaborative efforts, resources and communication between FDA and the State.

Possible Regulatory Actions Resulting From Positive Environmental Samples

Refer to the Regulatory Procedures Manual and Part V of the referenced Compliance Program. In some cases, based on the significance of the findings, voluntary correction may be the most appropriate action by the firm. When voluntary correction is not forthcoming, the Agency should pursue the routine regulatory procedures listed below to obtain compliance (refer to FMD-86 for classification):

Regulatory Meeting

Based on the consensus of the conference call between the District, ORA HQ, and appropriate Center, the District may request that the firm appear in the District Office to discuss the findings, the public health implications for the product, and to elicit a response from the firm to address the findings.

Warning Letter

Following the Regulatory Meeting (optional) and after a discussion with HQ, a Warning Letter may be drafted and submitted to the appropriate Center for review.

Voluntary Recall

When the investigational findings are egregious, the firm’s response to these findings are not appropriate to protect the public health, and/or a strong potential route of contamination of final product is determined to exist, i.e. product in the market presents a health hazard, a voluntary recall of the product by the firm may be expected.

FDA-Requested Recall

Although not usual in the absence of demonstrating specific product contamination, an FDA-requested recall could be considered in sufficiently urgent situations, normally involving a Class 1 health hazard evaluation. Refer to RPM Chapter 7.

Seizure/Injunction

When the firm’s response is inadequate to protect the public health and the firm refuses to conduct a voluntary recall, or an FDA requested recall will not be in a timely manner which would protect public health, a seizure and/or injunction should be considered.

Re-inspection

A re-inspection of the firm should be conducted within 6 months of initial regulatory action (regulatory meeting or Warning Letter) or much sooner if there is reason to believe there are serious ongoing deficiencies. This re-inspection should again include collection of environmental swab samples and verification of corrective action taken by the firm to eliminate harborage of the pathogen and any violative practices whereby the finished food product could become contaminated.

If you have any general questions on environmental sampling please contact Cindy Lewis, OFFO/FFPOB at 510-337-6866, or Cindy.Lewis@fda.hhs.gov.

Document History:

Versio n

Status* (D,I, R, C) Date Author Name and Title Approving Official Name and

Title

1.0 I 07/2009 NORMAN FOGG, FOOD

PROGRAM EXPERT, DFI

Michael Rogers, Director, DFI

2.0 R 03/22/2010 NORMAN FOGG, FOOD

PROGRAM EXPERT, DDFI

David Glasgow, Director, DDFI

3.0 R 06/2012 NORMAN FOGG, FOOD

PROGRAM EXPERT, DDFI

David Glasgow, Director, DDFI

4.0 R 10/2014 CINDY LEWIS, FOOD PROGRAM

EXPERT, OFFO/DFFPOI

David Glasgow, Director, DFFPOI

* - D: Draft, I: Initial, R: Revision, C: Cancel

Change History:

Version 2.0: Change in Glove instructions; added additional vendors/sources for sampling materials.

Version 3.0: All significant changes/revisions highlighted in text.

Version 4.0: All significant changes/revisions highlighted in text.

Attachment 1: Comments on Environmental Sampling from Dr. Don Zink, Senior Science Advisor, CFSAN

In the past, we have had extensive internal discussions regarding the optimal method for environmental sampling. One the one hand, it is exciting that the agency is making a major commitment to sampling certain food plant environments at risk of Salmonella contamination and this fact alone will cause the food industry to focus on Salmonella. I think the industry will respond by placing greater emphasis on their own sampling programs and plant sanitation and we must not lose sight of the immediate public health impact.

As to the question of optimal methods for environmental sampling, there are some things we are certain of and some things that ultimately require further study. Again, we must not let our concern about the unknowns delay or distract us from the otherwise important mission of launching this investigative effort. There is widespread consensus that it is more productive to take the largest possible environmental sample and this means that sampling with suitable hand sponges or sponges on a stick designed for this purpose are far preferable to using cotton or Dacron swabs. It is our expectation that the vast majority of environmental samples will be collected with either sponges on a stick or hand-held sponges. In some cases, it may be necessary to use a cotton or Dacron swab to collect a sample from an area too confined to use sponges. Also, it may be useful to use a cotton or Dacron swab as a tool to remove debris from cracks and crevices for sampling. In this case, use aseptic technique to break the cotton or Dacron head off the swab and use the remaining stick as a scraping or gouging tool, then use a sponge or swab to gather the material into a whirlpack or equivalent sterile bag.

Much discussion has been devoted to the finer aspects of the ideal buffer: DE broth, neutralizing buffer, letheen broth, etc. Buffer formulations vary greatly and theories abound as to why one is superior to another. However, there is no definitive peer-reviewed scientific study that looks at all of the options in parallel using real-world environmental samples. Of those studies that have been reported, you can easily raise valid questions about the experimental design, sample size and sample handling as variables in the final outcome. Clearly, it is important to refrigerate samples and get them to the laboratory as quickly as possible and analyze them immediately upon receipt at the laboratory. The buffer we have selected for use in the near term, DE Neutralizing Broth, is known to have worked well in the past to recover both Salmonella and Listeria from food processing plant environments. While no one can say with any certainty that it is the optimal broth for this purpose, neither can we say with any certainty that any other broth is quantitatively superior. The DE Neutralizing broth will allow us to recover Salmonella from food processing plant environments. The ingredients of DE Neutralizing broth have been considered by the agency and some ingredients have even been vetted through our Office of Food Additive Safety. The vast majority of samples we will collect in food processing plants will be collected from non-food contact surfaces. However, the DE Neutralizing broth should not present a risk of contamination even to food contact surfaces. Any residue of the ingredients in DE Neutralizing broth that might remain on a food contact surface is trivial and presents no risk of food product contamination. DE Neutralizing broth has been used by many in the food industry for environmental sample collection and has not been associated with instances of product contamination.

The successful conduct of the environmental sampling assignment will require a certain amount of ingenuity and resourcefulness on the part of our investigators. We need to hear what practices work best as we refine this process for the future. However, it is important to strictly follow the instructions for sample collection, transportation and analysis. I have no doubt that we will find positive results and be able to take appropriate regulatory action based on our findings.

Don Zink

Don L. Zink, Ph.D.

Sr. Science Advisor

FDA/CFSAN

Attachment 2: Sample Equipment Sources

3M Website: http://solutions.3m.com/wps/portal/3M/en_US/Microbiology/FoodSafety/ Phone: 1-800-328-1671

Catalog # Description HS10DE2G Hydrated-Sponge with 10mL DE Neutralizing Buffer with Gloves SSL10DE 3M™ Sponge-Stick with 10mL DE Broth SSL100 3M™ Sponge-Stick BPPFV10DE 3M™ Mini Flip Top Vial with D/E Broth RS96010DE 3M™ Swab-Sampler with D/E Neutralizing Broth BP556 or BP561 3M™ Swab-Tubes with Friction Cap

HARDY DIAGNOSTICS

Website: http://hardydiagnostics.com Phone: 1-800-266-222

K108 D/E Neutralizing Broth, 10 ml BS10DE1 D/E Neutralizing Broth, cellulose sampling sponge SRK45 EnviroTrans™, swab rinse kit, D/E Neutralizing Broth, 5ml with swab

NASCO

Website: http://www.enasco.com/ Phone: 1-800-558-9595

B01256WA D/E Neutralizing Broth, 10 ml B01245WA 18-oz. Whirl-Pak® Speci-Sponge® Environmental Surface Sampling Bag

WEBER SCIENTIFIC

Website: https://weberscientific.com/app/catalog/ Phone: 1-800-328-8378

Item # Description Various Denatured Alcohol (Ethyl Alcohol); 70% ethyl alcohol may also be purchased from local drug stores.

Various Alpet-E3 Plus Alcohol Hand Sanitizer Rinse (Best Sanitizers) Various Sterile Phosphate Buffer (for Kosher facilities) Various Sterile Saline (for Kosher facilities)

FISHER SCIENTIFIC

Website: http://www.fishersci.com Phone: 1-800-766-7000

Item # Description Various 70% Denatured ethyl alcohol; 70% ethyl alcohol may also be purchased from local drug stores.

http://solutions.3m.com/wps/portal/3M/en_US/Microbiology/FoodSafety/ http://hardydiagnostics.com/ http://www.enasco.com/ https://weberscientific.com/app/catalog/ http://www.fishersci.com/

Attachment 3: CFSAN Guidance - Environmental Sampling for Detection of Listeria monocytogenes

SAMPLE COLLECTION

DO Collect Samples From: DON’T Collect Samples From:

Moist/wet areas with standing water Dry, clean areas Direct food contact surfaces Employees – work shoes, hands etc Floors and related areas – Under floor mounted equipment, scales (floor and table mounted)

Hand wash or eyewash stations

Sanitizing foot mats – if disinfectant is not maintained this can be a good harboring source and point of transfer to other areas of the facility

Packaging materials – jars, lids, etc

Cleaning Equipment – automated floor cleaning equipment, brooms, mops, waste containers especially underside, etc

Raw agricultural products – raw peanuts etc or any food contact surface used exclusively for raw foods.

Air conveying equipment – pressurized air lines, air hoses, condensate from pressurized air lines, HVAC evaporators and evaporator condensate pans

Outside the plant – roof, parking lot, walkways, etc.

Product conveyors – cables, belts, joints, where product residue…