Inspections, Compliance, Enforcement, and Criminal Investigations
SUBCHAPTER 4.3 - COLLECTION TECHNIQUE
- 4.3.6 - ASEPTIC SAMPLE
- 22.214.171.124 - General Procedures
- 126.96.36.199 - Sampling Dried Powders
- 188.8.131.52 - Collecting Water Samples
- 184.108.40.206 - Sample Handling
- 220.127.116.11 - Controls
- 4.3.7 - ADULTERATION VIOLATIONS
- 18.104.22.168 - Field Examination
- 22.214.171.124 - Random Sampling
- 126.96.36.199 - Selective Sampling
- 188.8.131.52 - Sample Criteria
- 184.108.40.206.3 - Insect Contamination
- 220.127.116.11.4 - Bird Contamination
- 18.104.22.168.5 - Chemical Contamination
- 22.214.171.124.6 - Mold Contamination
- 126.96.36.199 - Abnormal Containers
- 188.8.131.52 - Microbiological Samples
- 184.108.40.206 - Collection of Environmental and Product Samples for Food Susceptible to Contamination with Pathogenic Microorganisms
- 220.127.116.11 - Samples for Viral Analysis
Sampling operations must be carried out using techniques that ensure the sample is representative of the lot, the sample of the product is in the same condition as it was before sampling, and that the collection technique does not compromise the compliance status of the lot.
It is your responsibility to collect your own samples using techniques and methods which will provide the most ideal sample, yet not be objectionable to firm management. This subchapter and the sampling schedules that follow, contain many sampling techniques, but not all. Your training and experience will enable you to become proficient in most sampling operations. However, in new or unusual situations it is your responsibility to use imagination and ingenuity in getting the job done and, if necessary, to consult with your supervisor.
Restore lots to their original condition. Do not leave partially filled shipping cases, short weight or short volume containers in the lot after sampling. Do not leave the lot in any condition, which might encourage pilferage, or make it unsalable.
When collecting from either full cases or bulk containers, replace sampled units by back filling from a container selected for that purpose. Avoid contaminating the back-filled units. If necessary, correct the contents declaration on the container(s) from which sampled to reflect the actual contents present. Refer to IOM 4.2.2 if the dealer objects to back filling because of company policy, different codes involved, or for other reasons. As a last resort, accede to the dealer's wishes and sample intact units, but record the facts in your regulatory notes and place a brief explanation on the C/R. Carefully re-close all containers and shipping cases. (Commercially available glues in spray cans or plastic squeeze-type bottles are an effective means of re-gluing cartons and cases without defacing with tape or other methods.) Re-cooper or reseal barrels and drums, re-sew bags, etc. If necessary, request use of the dealer's employees in helping to restore the lot, or arrange through the dealer to employ outside help. See IOM 18.104.22.168.
Identify each container from which units are taken with the date, your initials and the sample number, or you may complete and affix an FDA 2426, Examination Label, to each shipping case or bulk container sampled. For burlap or woven bags, the FDA 2426 may be glued to tags, and the tags attached to the bags. Should the dealer object to your identification procedure, attempt to reach a compromise (e.g., placing the ID in an obscure location, etc.). If the dealer still objects, accede to his wishes, but record the facts in your regulatory notes. Positive identification of the containers sampled is important if it becomes necessary to resample the lot(s), or if an embargo, seizure, or other action ensues. It also aids the dealer to differentiate between containers that have been opened by FDA as opposed to those opened by pilferage or torn opened by rough handling. It may be necessary to mark more containers than sampled to assure proper identification of the lot. This can be done by using the Examination Label, a handwritten ID or by using a rubber stamp.
Do not use industrial or permanent type markers on sample containers which allow penetration by ink. Many inks will penetrate to the product and act as a contaminant, interfering with the analysis. Water base markers will run when damp and must be covered with tape. See IOM 22.214.171.124 for identification techniques.
Do not permanently identify articles that are borrowed and will be returned to the dealer.
To determine sample size, first consult your assignment. If the assignment doesn't specify the sample size, follow the guidance in the applicable Compliance Program. The IOM SAMPLE SCHEDULE, should be used if the Compliance Program doesn't state the sample size. If none of these furnish the sample size, consult with your supervisor or the laboratory. Collect sufficient sample to allow for the FDA reserve portion and the 702(b) portion. See IOM 126.96.36.199 and IOM 188.8.131.52.
The following table represents the devices for which there are sampling instructions in Compliance Policy Guides:
|Clinical Thermometers||See CPG 335.800|
|Condoms||See CPG 345.100|
|Surgeons and Patient Exam Gloves||See CPG 335.700|
In addition to providing instructions on sample size, these compliance policy guides provide guidance on criteria to determine adulteration and whether or not regulatory action shoul be recommended. See WEAC’s webpage for additional guidance involving glove sampling.
When the sample schedule, assignment or other instruction does not specifically provide for the 702(b) portion, collect a sufficient amount to provide this required portion. You are not required to obtain a 702(b) portion in the following instances exempted by statute or by regulation 21 CFR 2.10(b):
- Devices are not included in the statutory requirement of Section 702(b).
- The amount available for sampling is less than twice the quantity estimated to be sufficient for analysis, in which case, collect all that is available.
- The cost of twice the quantity estimated to be sufficient for analysis exceeds $150.00. (Currently 21 CFR 2.10 uses $50.00 as the amount. However, ORA policy sets a limit of $150.00. If the sample is critical, and the cost exceeds $150.00, check with your supervisor.
- Import samples, collected from a shipment being imported or offered for entry into the United States.
- The sample is collected from a person named on the label of the article or his agent, and such person is also owner of the article. For example, it is not necessary to obtain a 702(b) portion if the sample is collected from a lot owned by and in the possession of the manufacturer whose name appears on the label.
- The sample is collected from the owner of the article or his agent, and the article bears no label, or if it bears a label, no person is named thereon.
Note: Regardless of the exemptions under 21 CFR 2.10(b) listed above, collect the 702(b) portion for filth samples unless your supervisor directs otherwise.
Whenever possible, collect separate subdivisions in order to provide the firm a portion as required by Section 702(b). Each duplicate subdivision should be collected from the same bag, box, case, or container. The total sample should be at least twice the quantity estimated to be sufficient for analysis, including a reserve portion for FDA's laboratory. If unable to collect separate subdivisions, assure that the total amount collected for each sample subdivision, or the total amount collected from an undivided sample, is at least twice the amount estimated to be sufficient for analysis. See IOM 184.108.40.206.
The exterior of any domestic package thought to contain an article subject to FDA regulation and in the possession, control, or custody of a common carrier may be examined (photographed, information on the outside copied, etc.) and records of the shipment may be obtained. Such package may not be opened either by an FDA employee or by an employee of the common carrier at the request of an FDA employee except as provided below.
The Office of Chief Counsel has advised FDA employees may, without a warrant, open, examine the contents and/or sample a package which is part of a domestic commercial interstate shipment in the possession, control, or custody of a common carrier only if:
- The consignor or consignee affirmatively consents to examination and/or sampling of the contents; or
- The Agency has reliable information the carrier regularly carries FDA regulated articles, and the facility where the sampling is contemplated is subject to FDA inspection. Reliable information may come from agency files, the carrier itself, other customers of the carrier, etc. and
- The Agency has reliable information a particular package sought to be examined is destined for, or received from another state, and contains an FDA regulated article. [Such information may be found on the exterior of the package and/or shipping documents in specific terms. Information may also come from reliable sources, which establish the consignor is in the business of manufacturing and/or shipping FDA regulated articles using a distinctive type of package (shipping container); and the package in question meets such description and shows the consignor to be such firm.]
Confer with your supervisor on any question concerning the need for a warrant. However, headquarters approval must be obtained because such inspection and sampling may require a search warrant. Contact the Office of Operations (OO) to discuss the matter. They will coordinate as necessary with Office of Enforcement and Import Operations and the Office Chief Counsel and provide further instructions.
If a decision has already been made by the district office to obtain a warrant, follow the procedures outlined in the Regulatory Procedures Manual, Chapter 6-3.
If a common carrier reports a violative article which it discovers under its own package opening procedures, independent of any request by an FDA employee or any standing FDA cooperative program with the carrier, FDA may still need a warrant to examine the material. Unless all the conditions for independent sampling in IOM 220.127.116.11 1 or 2 exist, you must consult with your supervisor, who will arrange for headquarters consultation as outlined above.
Note: Where the identity of an Interstate product is known by virtue of it being visible in bulk, or being in labeled containers or packages which are verified as to contents by shipping records, and where such product is under FDA jurisdiction at a given location, it may be sampled according to established IOM procedures.
If it is necessary to break the commercial seal to enter a railcar or other conveyance, reseal the door with a numbered self-locking "U.S. Food and Drug" metal seal. Record in your regulatory notes (and on C/R if sample taken) the number of the car or conveyance, the identifying number on any car seals removed, and the number of the FDA metal seals applied.
Do not collect human or animal biological materials (urine, feces, sputum, blood, blood products, organs, tissue etc.) unless arrangements for special handling and special treatment have been made in advance. Most ORA servicing laboratories are not prepared or certified to handle these materials. In addition to guidance for special sampling situations provided below, sampling guidance may also be found in IOM Subchapter 1.5 – Safety under IOM 1.5.3 - Sampling.
Sampling Containers for Lemon Oil or Other essential oils - Plastic or paraffin-coated liners in caps of containers used to hold samples of this type of product are not satisfactory in that the plastic or paraffin is soluble in the oils and interferes with the analysis. Use glass, cork, foil covered, or non-plastic, non-paraffin closures.
Sampling medicinal and other gases - Gases represent a special sampling situation. Please contact your servicing lab to determine an appropriate sampling container and sample size.
Detailed instructions for investigating and sampling products in connection with consumer complaints, tampering, foodborne outbreaks, injury and adverse reactions, etc. appear in the following subchapters and sections of the IOM:
IOM 8.2 - Complaints IOM 8.2.7 - Sample Collection IOM 8.3 - Investigation of Foodborne Outbreaks IOM 8.3.3 - Sampling Procedure IOM 8.4 - Investigation - Injury and Illness Reactions IOM 8.8 - Counterfeiting/Tampering IOM 18.104.22.168 - Sampling
Be cognizant of conserving scarce resources when investigating consumer complaints that do not involve injury, illness, or product counterfeiting / tampering. Unnecessary samples waste both operational and administrative resources. Use judgment as to whether or not it is necessary to collect the consumer's portion in situations that do not involve injury, illness, or product tampering. For example, there is little need to collect a physical sample of an insect infested box of cereal from the complainant. Both you and the consumer can readily see it is insect infested. The laboratory would find it insect infested, and the district would merely report the same thing back to the complainant. No practical purpose would be served by either collecting or examining such a sample.
See IOM 8.5.
If this type sample is desired your supervisor will provide specific instructions and procedures to be followed. This may involve:
- Whether to use your correct name or an alias. Caution: if you use an alias, do not use a similar name or a name with initials the same as yours (e.g., Sidney H. Rogers should not use Samuel H. Right). In addition, do not use a district office or resident post as a return address when ordering products or literature.
- Do not telephone your order in from the office or your home phone because the firm may have "Caller ID" and be able to identify your location by the phone number.
- Whether to use order blanks contained in the promotional package, advertisement, or promotional activity; or whether false ones will be used.
- Whether money orders, your credit card numbers, bank checks, or your personal checks should be used for payment. It depends on the situation, but money orders are preferred since these do not involve personal accounts.
- Where the requested items are to be sent: rented P.O. Box, home address, General Delivery, or other address.
- How the address and/or your name is to be recorded on the order blank. A code may be used either in your name or address so any follow-up promotional material sent to that name and address can be keyed to your original order.
When it has been decided to induce a sample and you have discussed the procedures with your supervisor, prepare the order and obtain the money order, or payment document. When all documents for ordering the item(s) are prepared, photocopy all the material, including the addressed envelope, for your record and submit the order.
When the order is received, identify the sample item, all accompanying material such as pamphlets, brochures, etc. (including all wrappings containing any type of printing, identification, numbers, post marks, addresses, etc.), and submit the item and exhibits in the same manner as any other official sample. If payment of the item was by personal check or credit card number, attach a photocopy of the canceled check or credit card receipt if available. You may do this later, after clearance of the check or charge slip.
See IOM 22.214.171.124.
Aseptic sampling is a technique used to prevent contamination by your sampling method. Aseptic sampling involves the use of sterile sampling implements and containers. Your sampling technique is where the lot or sample is contacted only by the sampling implements or the container. Samples collected using aseptic technique, will permit testimony that the bacteriological findings accurately reflect the condition of the lot at the time of sampling and, ideally, at the time of the original shipment. Whenever possible collect intact, unopened containers. Aseptic sampling is often used in the collection of in-line samples, environmental samples, product samples from bulk containers and collection of unpackaged product that is being collected for microbial analysis. Note: Products in 55 gallon drums, or similar large containers, either aseptically filled or heat processed, should not be sampled while the shipment is en route unless the owner accepts responsibility for the portion remaining after sampling. Try to arrange sampling of these products at the consignee (user) so the opened containers can be immediately used or stored under refrigerated conditions. Use ASEPTIC TECHNIQUE when sampling these products. For more guidance on aseptic technique, you may consult the course Food Microbiological Control 10: Aseptic Sampling, which is available to FDA employees through the ORA U intranet site.
If it is necessary to open containers, draw the sample and submit it under conditions, which will prevent multiplication or undue reduction of the bacterial population. Follow the basic principles of aseptic sampling technique. Take steps to minimize exposure of product, sampling equipment, and the interior of sampling containers to the environment.
Use only sterilized equipment and containers. These should be obtained from the servicing laboratory or in an emergency, at local cooperating health agencies. Pre-sterilized plastic or metal tools should be used. However, if unavailable, the metal tools can be sterilized immediately before use with a propane torch. Permit the tool to cool in the air or inside a sterile container before using. Soaking with 70% alcohol and flaming off is an acceptable method of field sterilization, and may be used as a last resort.
If it is necessary to drill, saw, or cut the item being sampled (such as large frozen fish, cheese wheels, frozen fruit, etc.), if at all possible, use stainless steel bits, blades, knives, etc. Wooden handled sampling instruments are particularly susceptible to bacterial contamination, are difficult to sterilize, and should be avoided.
Be extremely careful when using a propane torch or other flame when sterilizing tools and equipment. Evaluate the conditions pertaining to explosive vapors, dusty air, flame-restricted areas, firm's policy or management's wishes. The use of supportive devices should be considered when torch is not being hand held. Also be sure all flammable liquids, such as alcohol, in your filth kit are in metal safety cans and not in breakable containers.
If it is necessary to handle the items being sampled, use sterile disposable type gloves (rubber, vinyl, plastic, etc. - surgeon's gloves are good). Use a fresh glove for each sub and submit an unopened pair of gloves as a control. See IOM 126.96.36.199.
When opening sterile sampling containers, work rapidly. Open sterile sampling containers only to admit the sample and close it immediately. Do not touch the inside of the sterile container, lip, or lid.
Do not collect samples in areas where dust or atmospheric conditions may cause contamination of the sample, unless such contamination may be considered a part of the sample.
Cautions - The proper aseptic sampling of dried milk powder, dried eggs, dried yeast, and similar types of products is difficult because they are generally packed in multilayer poly-lined paper bags. These may be stitched across the entire top, may have filler spouts, or the top of the poly-liner may be closed or sealed with some type of "twists". The practice of cutting an "X" or "V" or slitting the bag and folding the cut part back to expose the contents for sampling should not be used because it creates a resealing problem; the opening cannot be properly repaired. The following procedures have been approved by the scientific units in Headquarters and should be used when sampling this type product.
- Remove as much dust as possible from the seam end by brushing and then wiping with a cloth dampened with alcohol. Note: This does not sterilize the bag as porous paper cannot be sterilized.
- Remove the seam stitching carefully (and dust cover, if any) and spread the walls of the bag and the poly-liner open enough to permit sampling being careful that no extraneous material such as dust, bits of twine, paper, etc., drops into the product.
- Carefully scrape off the surface of the product with a sterile device and aseptically draw the sample from the material below.
- Carefully reclose the bag and re-stitch by hand, or by machine if firm or FDA portable sewing machine is available.
188.8.131.52.2 - Bag Stitched Across Top And Poly-Liner Twist-Closed And Sealed With "Twist" Device - Wire, Plastic, Etc.
- Brush, alcohol wipe, and remove stitching as described.
- Remove "twist" seal and carefully open poly-liner using caution that no extraneous material drops into the product.
- Draw aseptic sample in same manner as in step 3 above.
- Carefully close the poly-liner with a twisting motion and reseal with "twist" seal arranging it so it will not punc-ture the poly-liner, and re-sew bag as in step 4 above.
The filling spout will be located at one side of the top stitching and will either pull out to form a top or side spout.
- Brush and alcohol wipe the area around the spout and carefully pull it out to reveal the opening. It is better to have the bag on its side while pulling the spout so any dust in the opening falls outside the bag.
- Carefully spread the sides of the spout apart and aseptically draw the sample. A trier or long handled device is usually better for this type opening because of the limited opening.
- Carefully close the spout with a firm twisting motion and be sure the opening is closed prior to pushing back into the bag.
When it is necessary to collect water samples for bacteriological examination, use the following procedures:
- Use sterile bottles. If dechlorination of sample is necessary, sodium thiosulfate sufficient to provide 100 mg/l should be placed in the clean bottles prior to sterilization. The sodium thiosulfate will prevent the chlorine from acting on the bacteria and assures, when the sample is analyzed, the bacterial load is the same as when collected.
- Carefully inspect the outside of the faucet from which the sample will be drawn. Do not collect sample from a faucet with leaks around handle.
- Clean and dry outside of faucet.
- Let the water run from the fully open faucet for at least 1/2 minute or for 2 or 3 minutes if the faucet is on a long service line.
- Partially close faucet to permit collecting sample without splashing. Carefully open sample bottle to prevent contamination, as for any other aseptic sampling operation.
- Fill bottle carefully without splashing and be sure no water from your hands or other objects enters the bottle. Do not over fill, but leave a small air bubble at top.
- Unless otherwise instructed, minimum sample size for bacteriological examination is 100 ml.
- Deliver sample to lab promptly. If sample is not examined within 24 hours after collection, the results may be inaccurate.
Note: When documenting specific situations in a plant, you may need to vary this procedure to mimic the actual conditions used by the firm.
For frozen samples, pre-chill sterile containers before use and keep frozen with dry ice. Use ordinary ice or ice packs for holding and transporting unfrozen samples that require refrigeration. See IOM 184.108.40.206, 220.127.116.11 and 18.104.22.168. Under normal circumstances dried products may be shipped unrefrigerated except in cases where they would be exposed to high temperatures, i.e., above 37.8°C (100°F).
Submit samples subject to rapid spoilage (specimens of foods involved in poisoning cases, etc.) by immediate personal delivery to the bacteriologist where feasible.
When collecting samples using aseptic technique and the subs are collected using pre-sterilized containers and equipment, submit a number of control subs. List control subs on your C/R. Control subs should be identified with a different nomenclature than the physical sample, i.e., a, b, c versus 1, 2, 3.
Examples of various control subs are:
- Sterile Containers - Where sterile containers are used to collect aseptic samples, submit one unopened container, which was sterilized in the same manner as containers used for sampling.
- Sterile Disposable Gloves - If sterile disposable gloves are used to handle the product, submit one unopened pair of gloves as a control.
- Sterile Sampling Equipment -Where presterilized sampling tools are used (e.g., spoons, spatulas, triers, etc.), submit at least one unopened sampling tool as a control.
Since adulteration samples are collected to confirm the presence of filth or other deleterious material, they are generally either larger or more selective than samples collected for economic or misbranding purposes.
When widespread evidence of filth or other adulteration is present, 402(a)(4) conditions can be documented by selective sampling. See IOM 22.214.171.124. For adulteration with filth, you will need to field examine (See IOM 126.96.36.199) a number of lots of product to determine the extent of the adulteration and can collect an investigational (INV) sample (See IOM 4.1.6) of filth exhibits and take photographs to document the widespread nature of the evidence. Collect separate sub samples of filth from various areas of the firm to illustrate the extent of adulteration within the firm. Field examine various lots of regulated products and collect official selective samples to document filth or other adulteration. Filth found on the exterior of containers, on pallets containing regulated product, or on the floor adjacent to lots of regulated product you are selectively sampling can be considered subsamples of that official sample. Consult with your supervisor and be guided by the criteria in Compliance Policy Guide (CPG) 580.100 Food Storage and Warehousing - Adulteration - Filth (Domestic and Import). The criteria in the Compliance Policy Guide can be used to determine if a particular lot meets the minimum criteria for direct reference seizure. Documenting a number of lots which meet the criteria helps establish the widespread nature of the adulteration.
When lots appear actionable, determine recent sales from the lot in question. Follow up may be necessary as directed by your supervisor.
Some field examinations are also referred to as bag-by-bag exams or unit by unit exams. When you conduct such exams take care to describe observations of each unit of product examined, any physical subsamples collected which reflect the violative nature of the lot, and exhibits which corroborate your report of observations. Record in your regulatory notes, subsequently in C/R Collection Remarks field or Continuation Form, or on Analyst Worksheet FDA 431, the results of your unit by unit examination of the lot. Observations should be specific. Report the general storage conditions, the violative condition of the lot, the physical relationship of the violative lot to other lots in the area, how you conducted the examination and how many units you examined. Wherever possible, record quantitative observations. Report the number and location of live and dead insects, rodent pellets, or other adulteration discovered inside the containers as well as on their exterior surface. Provide graphic measurements of areas of urine/chemical stains on each container and the extent of penetration. Correlate findings of the unit by unit examination with any photographs and physical subsamples collected. Where the field examination is carefully described and documented, the sample collected from obviously violative lots may be reduced to carefully selected exhibits. The field examination and the report of findings will serve as the analysis.
The concept of random "blind" sampling is to yield information about the average composition of the lot. It is employed when you have no information or method of determining which units are violative. Usually the violation is concealed and must be found by laboratory methods.
Sample size is usually described in your assignment, IOM Sample Schedule, Compliance Program Guidance Manual, or the applicable schedules. If none of these furnish the sample size, a general rule is to collect samples from the square root of the number of cases or shipping containers but not less than 12 or more than 36 subs in duplicate. If there are less than 12 containers, all should be sampled. Discuss sample size and 702(b) requirements with your supervisor. See IOM 188.8.131.52.
In some situations, random sampling is unnecessary or even undesirable. Under these conditions, examine the lot and select the portions which will demonstrate the violative nature of the lot. In addition to the selective samples collected, exhibits should include diagrams and photographs to demonstrate the violative conditions reported, and which containers were sampled and photographed.
The Agency has defined minimum direct reference seizure criteria to assist in assessing filth of individual lots. Criteria for rodent, insect, and bird filth are defined in Compliance Policy Guide (CPG) 580.100, Food Storage and Warehousing - Adulteration - Filth (Domestic and Import)for human foods, and reiterated in IOM sections 184.108.40.206 - 220.127.116.11. When collecting selective samples of products to show adulteration by filth, be guided by this criteria.
When evidence of rodent, insect, bird, or other animal activity is encountered during an inspection it is your responsibility to assess the evidence you observe and determine and document whether the activity is:
- Current or old
- Isolated to one lot (possible FD&C 402(a)(3) charges - contain in whole or in part filth or is otherwise unfit for food).
- Widespread, which requires evidence and documentation to illustrate all of the firm's susceptible products are potentially adulterated because they are being prepared, packed, or held under conditions whereby they may be contaminated. (possible FD&C 402(a)(4) charges)
Your assessment and documentation of the evidence observed (diagrams, photos and sample collections) will determine what actions may be required by either the establishment, the Agency, the Court, or all three to correct the problem. The evidence and documentation you collect and develop will be used to show, by a preponderance of evidence, that conditions at the firm have resulted, or could result in adulteration.
Your sample collection should be sufficient to document the extent of the violative conditions and not be limited to this minimum. Even where these minimum prerequisites are not met, you should collect samples as exhibits and evidence, particularly where adulteration under section 402(a)(4) of the FD&C Act [21 U.S.C. 342 (a)(4)] may be a factor. Your evidence may be used in a subsequent action against the firm, if corrections are not made.
Consult with your supervisor as soon as possible when you find evidence which meets the criteria set forth in CPG 580.100. If you are collecting several samples, the lab should be notified in advance that samples are on their way and should be analyzed expeditiously to facilitate regulatory action. Your supervisor may also want to notify your compliance branch so evaluation of evidence for a possible mass seizure can commence.
When Selective Sampling consists of an actual sample of a product, however small, as distinguished from bag cuttings, rodent pellets, insects, etc., a 702(b) portion must be obtained. In such cases, collect duplicate subs of the product to provide the 702(b) portion. This 702(b) portion is usually not an exact duplicate of the product collected for the Selective Sample, but should be collected from the same bag, box, or other container of product sampled. Whether collected from a container or bulk, the 702(b) portion should be taken as close as possible to that portion selectively sampled for analysis. Specify for each sub and duplicate collected, the origin, manner in which taken, and the examination to be made on your C/R. See IOM 18.104.22.168
Submit each portion of bagging or container portion, rodent pellets, material from beneath sampled area, control etc., in separate vial or subsample container.
It’s important when collecting a selective sample for adulteration violations that you:
- Use a coherent numbering/identification system for subsamples to avoid unnecessary confusion for the lab.
- Provide a detailed listing of individual sub descriptions on the C/R.
- If possible, provide a copy of any maps, photos or other additional documentation to the laboratory.
- Be sure to obtain product labeling. Since samples of lots which are sampled selectively are official samples, complete labeling must be collected. See IOM 4.4.9.
- Note: Whenever a portion of food is collected as part of a selective sample FD &C Act Section 704(d) applies and the CR should be marked as such.
The minimum direct reference seizure criteria to assist in assessing rodent adulteration of individual lots, as defined in Compliance Policy Guide (CPG) 580.100, are summarized as follows:
The storage facility is rodent infested and:
- Three or more of the bags in the lot are rodent gnawed; or
- At least five of the bags in the lot bear either rodent urine stains at least 1/4" in diameter, or two or more rodent pellets; or
- The food in at least one container in the lot contains rodent gnawed material, or rodent excreta or urine.
Whether or not the warehouse is rodent infested; IF:
- At least three bags bear rodent urine stains of at least 1/4" in diameter which penetrates to the product even though the product cannot be demonstrated to have been contaminated; or:
- At least two bags are rodent-gnawed and at least five bags bear either rodent urine stains at least 1/4" in diameter, with or without penetration to the product, or two or more rodent pellets; or:
- The food in at least one bag in the lot contains rodent-gnawed material or rodent excreta or rodent urine, and at least five bags bear either rodent stains at least 1/4" in diameter or two or more rodent pellets.
Additional regulatory guidance concerning rodent adulteration of pet foods can be found in CPG, 690.600 Rodent Contaminated Pet Foods.
Examine the exterior of the containers looking for rodent hairs, urine stains, excreta pellets, gnaw marks, holes, nesting material and live rodents. Make a diagram of the entire lot and note your findings as you examine the individual containers. You will need to include these descriptions on your C/R. Describe excreta pellets as carefully as possible, Note whether they appear dusty or shiny; soft or hard. Examine suspected urine stains with ultraviolet light in as near total darkness as possible. A minimum of 15 minutes is normally required for the eyes to become properly adjusted to accurately differentiate between rodent stain fluorescence and normal fluorescence of rice and certain other commodities. Wet, fresh or continually wetted runs may fluoresce poorly, but the odor of urine will usually be present and should be described on the C/R. Fresh dry urine stains will fluoresce blue-white, while older stains may be more yellowish/white. Rodent hairs will look like blue/white streaks. Look for the typical droplet pattern because rodents commonly urinate while in motion. Report the presence of droplet patterns on your C/R. Urine stained areas may be photographed under ultra-violet light conditions. Check with your supervisor about the technical aspects of this procedure. Do not mark container surfaces to outline the stained areas when taking either ultra violet or normal photographs. This may contaminate the product by migration through the containers. A number of things can interfere with the visual identification of urine stains. Many types of bagging and threading materials will fluoresce under U.V. light, however, the characteristic rodent stain fluorescence can be identified by its yellowish color and characteristic pattern. In addition a number of products exhibit a natural fluorescence. The following products may be difficult to evaluate because of either natural fluorescence or "quenching" of UV rays, even if contaminated. ("Quenching" refers to a covering up or a decrease in the ability of a product to fluoresce.)
|High Gluten Flour (Natural)||Burlap Bags (Quenching)|
|Nut Meats (Natural)||Bleached Sacks (Natural-White Glow)|
|Bean Flours (Natural)||Lubricants (Oils & Greases)|
|Brans (Natural)||(Natural-Blue/White to yellow/brown glow)|
|Pop & Field Corn (Natural)||Pitches & Tars (Natural-Yellow)|
|Wheat (Natural)||Detergents & Bleaches (Natural-White)|
|Starch (Natural)||Sulfide Waste Matter (Natural-Blue/White)|
|Spices (Natural or Quenching)|
Note clearly on your C/R if the product or package contains or is directly associated with any of the following:
- Dried milk products (contain urea).
- Whole grain wheat (contains urea and allantoin).
- Animal feeds (urea is usually intentionally added).
When sampling lots for rodent contamination follow the safety precautions in IOM 22.214.171.124. Wear gloves and handle the exhibits with tweezers or forceps. Handle exhibits carefully to prevent loss of microscopic evidence. Where you separate, count, or identify the various elements of an exhibit, (e.g.: sieve and find X number of rodent pellets), maintain the counted portions separate from the other subs. Note on the C/R those subs that were counted, separated, etc.
Collect a representative number of rodent pellets for laboratory confirmation. Place the pellets in a vial or other rigid container to prevent crushing. One of the identifying characteristics the lab looks for is the presence of rodent hairs in the pellets. The more pellets examined increases the possibility of a good identification. However, do not collect all the evidence you see as this would recondition the lot.
Collect portions of urine stains or gnawed holes from containers using small scissors or a sharp knife. Leave a portion of the stain or gnawed hole intact, but take a cutting large enough to provide good identification. Usually ½ inch around the stain is sufficient to allow manipulation during the lab exam. Note: The bag cutting should not be so large as to remove the entire contaminated portion, since this would recondition the product. For multilayer bags, be sure you cut through all layers of the bag and identify the layers with pencil. (Do not use ink as it often contains urea.) If possible, take stained cuttings from areas which have not been exposed for extended periods of time to light, in particular, ultraviolet light sources or to intense heat. If you have no alternative or cannot determine the stained areas' history, note the conditions on the C/R. Place cuttings and gnawed holes between 2 pieces of white paper, and then fold, roll, or leave flat and place into a glass container or other suitable container. This will hold the evidence in place and prevent possible loss of hairs or parasites due to static charges. Do not separate a multilayer cutting. Avoid the use of polyethylene containers as rodent hairs may adhere to containers made from this material. Put the cuttings in a large enough container to avoid excessive folding of the cutting.
Collect a minimal amount of product from under the stained area or hole, preferably just clumped product as a separate subsample. This prevents dilution of the contaminated product with uncontaminated product. Whenever you collect product, regardless of amount, collect a separate subsample to provide a 702(b) portion. See IOM 126.96.36.199.1. and identify per IOM 188.8.131.52.
Collect nesting material with minimal handling. A half cup is enough for analysis. Do not collect any rodents.
Product Control: In addition, you need to collect product controls, in duplicate, to provide for the 702(b) portion. These subsamples should be collected from beneath unstained portions of the container. Collect control samples from 3 different containers.
Packaging Control: Collect a portion of unstained container, which does not fluoresce, as a separate subsample for a control. As a general guide, collect the controls from the opposite side of the bag or make the cutting large enough to separate the control area and the stain. Separate the controls from the stains and submit in separate containers. Collect at least 3 container controls for each sample. If the lot consists of different containers or bags of different manufacturers, collect controls to represent each type or manufacturer of the containers.
Submit each portion of bagging or container, pellets, material from beneath sampled area, control, etc., in separate vial or subsample container. Place the subsamples in a dark container, such as a cardboard box to protect them from light and protect the exhibits from being crushed.
The complete official sample will consist of:
- Subsamples of rodent excreta pellets
- Subsample of nesting material
- Subsamples of stained bagging, or portions of the containers, and any adhering pellets.
- Subsamples of unstained bagging, or portions of the containers, which do not fluoresce, for controls (minimum three required).
- Subsamples of small portions of the product from directly beneath the stained areas. Do not dilute the contaminated product beneath the stain with the non-contaminated product.
- Subsamples of small portions of product to serve as 702(b) portions
- Subsamples of uncontaminated product from beneath the unstained bagging, or other container. These serve as controls, and should be collected in duplicate to provide 702(b) portions. Collect control samples from 3 different containers.
- Subsamples of cuttings from gnawed holes
- Subsamples of small amounts of product collected from beneath the gnawed holes.
- Subsamples of small portions of product to serve as 702(b) portions.
- Product labeling.
- Interstate documentation.
If conditions warrant, consider collecting an INV sample per IOM 4.1.6. to document widespread rodent activity.
The criteria from CPG 580.100 below, involving dead insects only, will not be used for action against any food intended to undergo further processing that effectively removes all the dead insects, e.g. processing of cocoa beans.
- The product contains:
- One live insect in each of two or more immediate containers; or, one dead insect in each of three or more immediate containers; or, three live or dead insects in one immediate container; plus
- Similar live or dead insect infestation present on, or in the immediate proximity of, the lot to show a 402(a)(4) [21 U.S.C. 342 (a)(4)]violation.
- The product contains one or more live insects in each of three or more immediate containers.
- The product contains two or more dead whole insects in at least five of the immediate containers. Note: a situation such as this may follow fumigation of the lot and vacuuming of the exteriors of the bags.
- The product is in cloth or burlap bags and two or more live or dead insects are present on at least five of the containers. Note: Some live insects must be present. Product need not be shown to have become contaminated.
Examine the exterior of the containers (especially along seams or creases) looking for insects, larvae, webbing, nesting material, entrance or exit holes, and cast skins. Make a diagram of the entire lot and note your findings as you examine the individual containers. Describe insects or larvae carefully, noting if they are dead or alive. You will need to include these descriptions on your C/R.
Collect a representative number of insects for laboratory confirmation. Consider the use of a moistened artist brush to collect subsamples. Place the specimens in a vial or other rigid container to prevent crushing. Collect all forms of insects you see, however do not collect all the evidence from the lot or you might recondition the product. If you collect live insects, be sure to note that on your C/R. However, you should not send live insects to the lab. Freeze the subsamples prior to shipment to ensure they are not alive when you ship them. Note the fact that the subsamples were frozen on the C/R. Cut portions of bags or containers containing suspected insect entrance or exit holes from containers using small scissors. Usually ½ inch around the holes is sufficient to allow manipulation during the lab exam. Note: The bag cutting should not be so large as to remove the entire contaminated portion, since this would recondition the product. For multilayer bags, be sure you cut through all layers of the bag and identify the layers with pencil. (Do not use ink as it often contains urea.) Place cuttings between 2 pieces of white paper, and then fold, roll, or leave flat and place into a glass container or other suitable container. This will hold the evidence in place and prevent possible loss microscopic evidence due to static charges. Do not separate a multilayer cutting. Avoid the use of polyethylene containers as insect fragments may adhere to containers made from this material. Put the cuttings in a large enough container to avoid excessive folding of the cutting Collect product from beneath holes which penetrate the packaging as a separate subsample. Whenever you collect product, regardless of amount, collect a separate subsample to provide a 702(b) portion. Note on the subsample itself and on your C/R which subsamples are the 702(b) portions.
The complete official sample will consist of:
- Subsamples of insects, larvae, webbing, etc.
- Subsamples of portions of the containers with entrance or exit holes.
- Subsamples of small portions of the product from directly beneath holes.
- Subsamples of small portions of product serve as 702(b) portions See IOM 184.108.40.206.1.
- Product labeling.
- Interstate documentation.
If conditions warrant, consider collecting an INV sample per IOM 4.1.6. to document widespread insect activity.
Per the criteria from CPG 580.100, if the product is in permeable containers (paper, cloth, burlap, etc.), and
- The product contains bird excreta in one or more containers, and you feel the insanitary storage conditions will clearly support a 402(a)(4) [21 U.S.C. 342 (a)(4)] violation.
- Bird excreta is present on the exteriors of at least five of the containers, and the product contains bird excreta in one.
- At least 30% of the number of bags examined, but at least five bags, are contaminated with bird excreta; and at least three of the bags bear excreta stains which penetrate to the product, even though the product may not be contaminated.
Note: In all instances of bird excreta contamination the excreta must be confirmed by positive test for uric acid.
Examine the exterior of the containers looking for bird excreta. Make a diagram of the entire lot and note your findings as you examine the individual containers. You will need to include these descriptions on your C/R.
Remove portions of bird excreta stains from containers using small scissors. Leave a portion of the stain intact, but take a cutting large enough to provide good identification. Usually ½ inch around the stain is sufficient to allow manipulation during the lab exam. Note: The bag cutting should not be so large as to remove the entire contaminated portion, since this would recondition the product. For multilayer bags, be sure you cut through all layers of the bag and identify the layers with pencil. (Do not use ink as it often contains urea.) If possible, take stained cuttings from areas which have not been exposed for extended periods of time to light, in particular, ultraviolet light sources or to intense heat. If you have no alternative or cannot determine the stained areas' history, note the conditions on the C/R. Place cuttings between 2 pieces of white paper, and then fold, roll, or leave flat and place into a glass container or other suitable container. This will hold the evidence in place and prevent possible loss of microscopic evidence due to static charges. Do not separate a multilayer cutting. Avoid the use of polyethylene containers as bird excreta may adhere to containers made from this material. Put the cuttings in a large enough container to avoid excessive folding of the cutting.
Collect a minimal amount of product from under the stained area, preferably just the clumped product as a separate subsample. This prevents dilution of the contaminated product with uncontaminated product. Collect a separate subsample to provide a 702(b) portion (See IOM 220.127.116.11.1).
Product Control: In addition, you need to collect product controls, in duplicate, to provide for the 702(b) portion. These subsamples should be collected from beneath unstained portions of the container. Collect control samples from 3 different containers.
Identify the 702(b) subsamples, as such on subsample identification (See IOM 18.104.22.168.) Note on the subsample itself and on your C/R which subsamples are the 702(b) portions.
Packaging Control: Collect a portion of unstained container as a separate subsample for a control. As a general guide, collect the controls from the opposite side of the bag or make the cutting large enough to separate the control area and the stain. Separate the controls from the stains and submit in separate containers. Collect at least 3 container controls for each sample. If the lot consists of different containers or bags of different manufacturers, collect controls to represent each type or manufacturer of the containers.
The complete official sample will consist of:
- Subsamples of stained bagging, or portions of the containers.
- Subsamples of unstained bagging, or portions of the containers for controls (minimum three required).
- Subsamples of small portions of the product from directly beneath the stained areas. Do not dilute the contaminated product beneath the stain with the non-contaminated product.
- Subsamples of small portions of product to serve as 702(b) portions.
- Subsamples of uncontaminated product from beneath the unstained bagging, or other container. These serve as controls, and should be collected in duplicate to provide 702(b) portions. Collect control samples from 3 different containers. Submit each portion of bagging or container portion, pellets, material from beneath sampled area, control, etc., in separate vial or subsample container.
- Product labeling.
- Interstate documentation.
Collect samples from lots suspected of dry chemical contamination in much the same manner as described for rodent urine. After collecting a sample of the contents from immediately beneath the suspected area, collect residues from the surface of the bag or container. In the case of infiltration of loosely woven bags, shake or tumble the bag over a large sheet of clean paper to collect the siftings as a sample.
The USDA/FGIS has approved a number of commercial screening tests for detecting aflatoxin contaminated corn. However, these tests usually require a chemical extraction process and are therefore not amenable to FDA field examination procedures.
The blacklight test (also referred to as the Bright Greenish-Yellow Fluorescence (BGYF) test) is a presumptive test used to screen and identify corn lots that should be tested further for aflatoxins. The test is based on BGYF observed under long wave (366 nm) ultraviolet (UV) light produced by the molds Aspergillus parasiticus and A. flavus on "living" corn (i.e. corn that has been stored less than 3 months). The growth of these fungi may result in aflatoxin production. Aflatoxins per se do not produce BGYF under long wave UV light. It is thought the BGYF is produced by the reaction of kojic acid formed by the fungi and a peroxidase enzyme from living corn. Corn that has been in storage for a lengthy period of time (3 months or more) may give false positive BGYF. Therefore, determine how long the corn being sampled has been in storage. If it has been in storage over three months, do not use the following field screening procedure.
Essential steps for this blacklight procedure are:
- A 10 lb. sample representative of the corn lot must be obtained by probing, or by continuously sampling a grain stream.
- Examine using a 366 nm UV light (portable black-lights meet this criteria).
- Wear goggles or use a viewer that screens out UV light. Shine the light on the corn sample which has been spread in a single layer on a flat surface in a darkened room.
- Use a 2 lb. portion, and carefully observe the entire corn surface one kernel at a time. Examine the entire sample using this procedure.
- Count all BGYF glowers (kernels or particles that "glow" bright greenish-yellow). Compare the BGYF color with a fluorescent standard, if one is available. Remember normal corn, if it fluoresces, will fluoresce a bluish white.
- If four (4) or more BGYF particles are detected in the 10 lb screening sample, collect a sample for laboratory analysis.
See IOM SAMPLE SCHEDULE CHART 2 - Sampling Schedule for Canned and Acidified Foods for listing can defects.
During inspections of firms producing products susceptible to microbial contamination (e.g., peanut butter, dried milk, dairy products, frozen ready-to-eat seafood,crème filled goods, breaded items, prepared salads, etc.), sampling may be warranted, based on observations or as directed in the Workplan, Compliance Program, or assignment. Proof of adulteration with fecal organisms, elevated levels of non-pathogenic microorganisms, or presence of pathogenic microorganisms must be established. Follow instructions under IOM 22.214.171.124 when collecting microbiological samples to document manufacturing conditions conducive to adulteration.
Mold Samples - During inspections of manufacturers such as canneries, bottling plants, milling operations, etc., it may be necessary to collect scrapings or swabs of slime or other material to verify the presence of mold. The sample should represent the conditions observed at the time of collection and consist of sufficient material to confirm and identify mold growth on the equipment. If possible, take photographs and obtain scrapings or bits of suspect material. Describe the area scraped or swabbed, e.g., material was scraped or swabbed from a 2" x 12" area. Suspected filth, collected from ceilings, walls, and equipment, for mold examination must be kept moist by placing it in a container with a small amount of a 3-4% formalin solution. Large amounts of slime may be placed in a wide mouth glass jar with either a 1% formaldehyde solution or a 3-4% formalin. Note: Formalin is normally sold as a standard stock solution of 37%. To obtain the required 3-4% formalin solution, mix 5 ml of the 37% stock solution with 95 ml of distilled water. This will yield the appropriate strength solution necessary to fix the mold. Although formaldehyde or formalin are the preservatives of choice you may preserve the subs in either a 50% alcohol solution or in acetic acid (full strength vinegar) if formaldehyde or formalin are not readily available. The above instructions apply to the collection of raw material, in-line and finished product samples for mold. However, in-line and finished product subs such as doughs, etc., which may be harmed by the formaldehyde, may be frozen. Check with your laboratory for its recommendation regarding preserving mold samples.
126.96.36.199 - Collection of Environmental and Product Samples for Food Susceptible to Contamination with Pathogenic Microorganisms
Sampling for products susceptible to microbial contamination and the environment in which they are produced may help identify the presence of pathogenic microorganisms before they can cause illness. With the recent increase in foodborne outbreaks and inspections identifying links between outbreaks and environmental (including non-food contact surface) contamination, there will be an increased focus on routine environmental sampling during inspections. Conduct environmental surface sampling as directed by the work plan, compliance program or assignment, or based on inspectional observations. If you are unsure of under what circumstances to perform environmental sampling, consult with your supervisor. Also see IOM 188.8.131.52 for inspectional guidance for firms producing products susceptible to contamination with pathogenic microorganisms. A microbiological inspection requires a thorough understanding of critical factors associated with the production of the specific product being inspected. To prove the establishment is being operated in an insanitary manner it is necessary to show the manufacturing operation or conditions at the facility are likely to, or have contributed to the bacterial load of the product. When feasible, inspections should cover equipment condition before a day's production begins and the clean-up at the end of the day's production. For environmental Salmonella sampling, it is preferable to sample before the plant conducts a wet cleaning operation. Sampling should include environmental sponges or swabs of food contact surfaces (particularly for Listeria monocytogenes) and non-food contact surfaces (particularly for Salmonella serotypes), based on observations, or as directed. In-line sampling should be conducted based on observations or as directed. Collect finished product only after consultation with CFSAN, HFS 605 Division of Enforcement, or as directed in the compliance program or assignment. When conducting microbiological inspections, whenever possible, an investigator/ microbiologist team approach should be used. For environmental sampling, a third person is recommended to assist with collection and/or record information.
CFSAN has developed guidance on the specific locations within a firm to collect environmental samples to increase the likelihood of detecting Listeria monocytogenes and Salmonella. See IOM Exhibit 4-20 and 4-21 and FIELD BULLETIN #30 – FOOD PROGRAM AREA INSTRUCTIONS FOR ENVIRONMENTAL SAMPLING for guidance on environmental sampling/locations for these microorganisms. In addition, please view the training video, "Environmental Sampling" which provides technical and procedural information on environmental sampling.
Environmental sampling, particularly for Salmonella, should be considered at manufacturing plants that are typically dry environments where water occasionally wets the area, either intentionally as part of a periodic wet cleaning or inadvertently as the result of leaking pipes or valves or a leaking roof.
At a minimum, collect from 100 up to 300 subsamples per production area in order to maximize the chance of finding the niche of the microorganism. For Listeria environmental swabbing, collect at least 50 swabs/subs and ideally 100 or more subs if presented with a sufficient number of promising sample sites. Salmonellae tend to be more difficult to detect in a contaminated plant vs. Listeria, and a greater number of samples are needed for Salmonellae environmental sampling in order to have confidence in swab findings/results.
In most cases, subsamples for Salmonella will be collected from the Zones 2 – 4 (see below), concentrating primarily on Zone 2. Samples should be collected from the equipment itself, particularly equipment mounting and support structures. When targeting Listeria, swabs will be collected primarily from Zones1 and 2.
"Environmental" sponge or swab sampling does not give quantitative results. Because a sponge or swab takes a very small sample, microorganisms of significance are often missed. It is important to keep in mind a negative result on a sponge or swab could negate an inspectional observation that is not fully documented. A positive finding will give more support to a fully documented observation. The environmental sampling zone concept should be utilized in determining where to conduct environmental sampling:
- Zone 1: Refers to all direct food contact surfaces such as slicers, mixers, conveyors, utensils, racks, work tables, etc. For inspections focusing on the presence of Salmonellae, such as firms producing peanut products and other dry product environments, food contact surfaces are normally not sampled unless specifically requested in the assignment or CP. In contrast, for inspections focusing on detection of Listeria monocytogenes, such as firms producing seafood or cheese products in a wet environment sampling of food contact surfaces is essential.
- 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 directly affect safety of 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.
- Zone 3: The area immediately surrounding Zone 2. Zone 3 is an area which, 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 cross-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.
A large majority of the environmental samples collected should be taken from Zones 1 (when directed and depending on the organism in question) and 2, and to a lesser degree Zone 3 areas. Very few, if any, environmental samples should be taken from Zone 4 areas. Document the possible link between the source of an environmental sample and contamination of the food product using both written descriptions and photographs. Describe the location of the sample in relation to areas where food is exposed and any mechanical or human activities you observe that might cause an organism to be spread beyond this niche environment. The district’s response to a positive swab will depend on the proximity of the sample location to the processing line and the likelihood of contamination contacting the product. For example, if an environmental sample (sponge or swab) was taken from: A potential Salmonella harborage niche beneath a piece of food handling/processing equipment – does the firm periodically wet clean the floor (which could cause the organism to be sprayed onto exposed food?) On the floor in a corner of the production room – Did you observe activity that could cause the organism to be picked up by employee’s shoes, pallet mover wheels, forklift wheels, or could get onto the hands of employees who handle the equipment? The ceiling area – Is condensate, flaking paint, etc., located over the processing area? Did you observe the condensate dripping on the food surfaces of the processing equipment and/or product? Correlate these observations with the potential to contaminate the food processing equipment or product. On occasion firms may opt to collect their own swabs in conjunction with your sample. If this occurs, request the firm to provide their results to you once known.
184.108.40.206.2 - Environmental Sampling Equipment And Instructions For Large And Small Area Environmental Surface Sampling
These instructions should be followed in order to ensure standardization of FDA environmental sample technique across districts.
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 survive the transport to the laboratory.
Day and Engley (D/E) neutralizing broth or buffer (the terms broth or buffer are used interchangeably for this product) 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 Office of Regulatory Science (ORS), is the one to be used for general purpose environmental sampling.
For large area environmental sampling hand held sponges or sponges on a stick should be used. The sponges on a stick reduce manual contact with the sponge during the sampling procedure and are good for accessing tight spaces. Dacron tip swabs are recommended for small area environmental sampling (approximately 10cm x10 cm, or 4 x 4 inches).
Sampling Equipment: If sources cannot be located for sponges or swabs prehydrated with D/E Neutralizing buffer or broth, use unhydrated sponges and swabs along with single use tubes of D/E neutralizing broth. Other types of hydrated sponges and swabs that contain either a neutralizing broth or an enrichment broth but not both are not suitable. Addition of D/E broth to these may dilute the concentrations of both components to the extent they will not be effective.
Hand held sponges or sponge on a stick pre-hydrated with D/E neutralizing broth if available, dry hand held sponges or sponge on a stick, swabs pre-hydrated with D/E neutralizing broth, dry swab in swab tube with screw on cap or single use tubes of D/E Broth are recommended.
If you need sourcing information for equipment please contact Division of Food and Feed Operations and Inspections at (301) 796-0360.
Other general sampling supplies you will need for environmental sampling:
Sterile gloves (size 7 and 9 to include latex free styles) Hand sanitizers (wash and sanitize hands often during sampling) Cooling medium for samples Boxes or coolers Labels to ID samples Permanent marker Flashlight Sterile metal spatulas (small) or other sterile implement to scrape debris out of cracks
It is important to use sponges or sponges on a stick for the large majority of samples since you can sample and “scrub” a larger area with a sponge compared to a swab. Swabs are only appropriate for areas that are inaccessible to sponges.
When collecting environmental samples you should make every effort to collect as much sample material as possible from the processing plant environment in order to maximize the chance of finding low numbers of organisms.
For large area environmental sampling, hand-held sponges or sponges on a stick should be used. The sponges on a stick reduce manual contact with the sponge during the sampling procedure and are good for accessing tight spaces. Dacron tip swabs are recommended for small area environmental sampling (approximately 10cm x10cm, or 4 x 4 inches) and for cracks and crevices.
For collection of environmental samples in Zones 2 - 4 and for firms targeted as part of routine surveillance inspections only, it is not necessary to change gloves between each sub provided that the CSO or analyst 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 swabs are collected in Zone 2, the CSO or analyst would not need to change gloves between each of these subs until 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 isopropyl alcohol. It is expected that collection of a large number of subs in one area would necessitate several changes of gloves.
For swabs collected in Zone 1 and during “for-cause” inspections (such as those conducted in response to a current or previous outbreak, or an emergency), continue to follow the established policy and change gloves between each sub as described in the Environmental Sampling training video.
Sampling of Dry Surfaces:
Using a felt-tip black permanent marker, 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. From the outside of the sponge bag manipulate the handle toward one side. Pull off the top of the whirl-pak bag holding the Sponge-stick along the perforation. Using the tabs on both sides of the wired band, pull gently to open the 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 D/E 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 if preferred.
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 if 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 buffer to keep the sponge gliding smoothly over the surface. If there is excess buffer, squeeze it back into the whirl pack bag and continue until you have sampled the entire sampling site.
7. After sampling, return the sponge to original Whirl-Pak bag with any excess buffer, snap off the handle in accordance with the product instructions that accompany the Sponge-stick, and submit as a subsample.
8. Remove the used sterile glove and discard.
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 Whirl-Pak 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.
Sampling of Wet Surfaces:
Sample using aseptic techniques with a dry Sponge-stick following the general instructions above for removing the Sponge-stick from the bag, and for swabbing. After sampling, return the Sponge-stick to the original sterile Sponge-stick bag and using aseptic techniques add 10 ml of D/E neutralizing broth to the bag. Proceed as instructed in #5-10, above.
Small Area Environmental surface sampling procedure (approximately 10cm x10cm, or 4 x4 inches):
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 pre-hydrated with D/E Neutralizing Solution. 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 vial, place in a Whirl-Pak bag, and as soon as possible place inside an insulated cooler with pre-frozen gel pack for transport/shipment to the laboratory. Dust and debris scrapings may also be collected using a sterile implement from facilities producing dry products such as nuts and powders. A minimum of 5 to 10 grams should be collected with 100 grams being optimum. When sampling mops or brooms, swabbing with a sterile sponge pre-hydrated with D/E Neutralizing solution is an efficient method although mop strands and broom bristles may also be clipped and submitted.
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 solution to the swab and transport to laboratory as noted above.
Collect debris on equipment and from floor defects, joints and gaps. Debris can be scraped out using a sterile implement, such as a small metal spatula. A minimum of 5 to 10 grams should be collected, with 100 grams being optimum.
For environmental samples only, collect one closed control for each distinct lot of sterile equipment used and submit with the final collection of subs on the last day of sampling.
Open controls are not to be submitted for environmental sample collections.
Often multiple days are required to collect an appropriate number of environmental swabs. For each day of collection during an inspection, assign a new sample number. However, the subs should continue in sequence throughout the collection period, i.e. day 1, Sample # 111111, subs 1 – 100, day 2, sample # 111112, subs 101-175. Link the sample numbers to the assignment for tracking purposes. Environmental swab subs should be numerical, i.e. 1, 2, 3, etc.; control subs should be alphabetic, i.e. a, b, c, etc.
All environmental samples, including swabs, soil, water, and animal scat, are to be identified as Investigational (INV) under product code "52YYY99", Do NOT use the product code of the covered product for environmental samples.
In-line sampling should be conducted as directed or based on inspectional observations.
Each in-line subsample will consist of approximately 114 g (4 oz), in duplicate (702(b) portion), if that amount is available (Also see IOM 220.127.116.11 - 702(b) Requirement). All in-line samples must be collected aseptically. Sampling Areas (this is not a comprehensive listing of areas to collect in-line samples, since each firm will be different, depending on processing/packaging techniques and the finished product produced: "Raw" ingredients used in the manufacturing of finished foods (including those conveyed by bulk tankers) should be considered for sampling to determine the effect of subsequent processing on bacterial content. Of particular concern are raw materials which can support microbial growth, are not normally cooked or prepared in a manner lethal to pathogenic microorganisms (such as dairy, soy, corn or sugar syrup based products), and adequate controls to ensure the safety of the finished product are not in effect. Since the major portion of some finished food products are not homogeneously contaminated, it may be necessary to collect multiple subsamples of the raw material(s) to establish a reliable microbial base line. Obtain sequential subsamples with the view of bracketing each step of the processing operation, in particular those steps suspected as routes of product contamination. A series of in-line samples should be collected during the first part of a shift, and a duplicate series during the latter part. If products or components are heated (e.g., blanched, boiled, etc.) take subsamples immediately before and immediately after heating, before possible insanitary equipment and processing delays contribute to bacterial increases. Particular attention should be given to determine routes of cross-contamination from the raw product to the "heated" product, especially if this heating step is critical to the destruction of pathogenic organisms. If a product is capable of supporting microbial growth and is not being handled expeditiously, sample before and after this particular processing step. Take time and temperature measurements of cooking, freezing and cooling procedures. Sample when appropriate to demonstrate possible microbial growth. Large masses of ingredients may cool or warm slowly enough to permit microbial growth. Improperly cleaned equipment may contaminate the product with bacteria. This may result in either a uniform or a spotty increase in bacterial numbers. If possible, scrapings of questionable material should be in sufficient quantity to be easily weighed and quantitatively diluted, if collected for analysis.
Collect finished product as directed in the compliance program, assignment or by your supervisor. Collect product from production on the day of the inspection and from the previous day's run. Sampling multiple lots should be considered depending on the type of product and process used. The subsamples should consist of ten (10) retail size containers at least 114g (4 oz) each, in duplicate (702(b) portion).
If the finished product is also to be analyzed for Salmonella, collect samples in accordance with instructions in the IOM. See Salmonella Sampling Plan, Schedule Chart 1.
Environmental sampling in the foods program has had increasing focus in assignments issued to the Field. FDA/ORA, with the concurrence of and in conjunction with Office of Chief Counsel (OCC) and the ACRA, has outlined criteria in order to implement a consistent policy for the reporting of positive environmental sample results on the FDA 483 as applicable to the foods program only. Current policy, going forward, is to report significant positive environmental sample results, from swabs collected at food firms, on the FDA 483, if the results are known prior to the conclusion/closeout of the inspection. In addition, districts are not being asked to unnecessarily extend inspections to include these results. Reasoning behind the implementation of this policy includes:
- Informing the firm of positive results where food products are concerned
- Eliciting firm feedback in response to positive results
- The opportunity to provide relevant information to both regulators and the public when released under FOIA thereby potentially uncovering and linking other investigational information that can aid in the determination of root contamination cause(s)
- The responsibility to document positive environment sample results as significant observations that can contribute to potentially unsafe conditions as they pertain to the Public's health.
Positive environmental sampling results should be noted on the FDA 483 when the following conditions are met:
- Related to a current or future foods program inspection/investigation
- Inspection has not been closed (Note: it is not requested that the period of inspection be extended for the purpose of receiving analysis results)
- Positive sample finding(s) is/are a significant observation, i.e. a route of contamination from the environment to the product is clearly demonstrated, such as, for example, positive sample result(s) in Zones 1 and/or 2 for Listeria or positive sample result(s) in Zone 2 and/or 3 for Salmonella
Findings in Zone 3 (Listeria) and Zone 4 for either pathogen should not be reported on the 483 as they are normally not considered significant, except in combination with positive findings in Zones 1 or 2, when these would further strengthen regulatory action.
Sample instructions will be issued by the appropriate Center on a case by case basis.
Field weighing for net weight is primarily to determine the likelihood of short weight units. The laboratory will confirm both tare and net weights. Use a Gurley, Troemner, or equivalent balance. Check the accuracy of the balance before and after use. If this equipment is not available, or the units exceed their capacities, use commercial scales. If possible, have the commercial scales checked in your presence by the local Sealer of Weights and Measures. If this is not possible, report the name, type of scale, style and capacity, minimum graduations, apparent sensitivity, and date of last sealing and by whom.
Whenever possible, determine a minimum of six tares selected at random. If empty containers are readily available, or if tares vary widely (e.g.; glass jars), determine at least 12 tares.
Weigh 48 units, if that number is available, selected at random from the square root of the number of cases in the lot with a minimum of 6 and a maximum of 12. Where units are selected from the production line, do so in representative manner. Report the code weighed and if short weight, the quantity in the code. Unless otherwise instructed, do not weigh leaking containers. Identify each unit with the corresponding sub number on the Field Weight Sheet (FDA 485). Submit the units indicated by the asterisks on the FDA 485 plus twelve additional weighed units for reserve if the average net is below that declared on the label.
Record weights on Form FDA 485, Field Weight Sheet. See IOM Exhibit 4-6. Submit Field Weight Sheet with the printed FACTS Collection Record.
Individual Captions: Block 1 Date - Enter the date weighed. Block 2 Sample No.- Enter the sample number of the C/R. Block 3 Product - Enter the specific name of the product, i.e., macaroni in cellophane, butter in aluminum wrappers, olive oil in glass, etc. Quote significant portions of the label including the declared net weight. Block 4 Type of Balance - Enter the type of balance used i.e., Gurley, Troemner, etc. If balance used is not FDA equipment, give style, capacity, minimum graduations, etc. Block 5 Responsible Firm and Address - Enter the name and address of the firm most likely responsible for the short weight violation. Block 6 Address Where Weighed - Enter the name and address or location where weighed. Block 7 Warehouse - Enter the type of warehouse where product is stored, i.e., cold storage, truck dock, production line, etc. Enter the temperature and estimate the humidity where possible. Block 8 No. Of - Enter the number of cases, and number and size of units per case in the lot. Enter the number of cases from which subs were weighed and the number of subs weighed from each case. If the units are collected from a production line, estimate the number of units produced of the code weighed. Block 9 Gross Weight - Arbitrarily assign and record the shipping case number from which each sub was weighed. Number each unit submitted to correspond with the sub number on the Field Weight Sheet. Record weights to second decimal place.
Block 10 Preliminary Tare - Determine and record tare weights as provided in IOM 18.104.22.168.1. Obtain the preliminary average tare by totaling preliminary tares and dividing by the number of tares weighed. Block 11 Weighing Results - Determine the average gross weight by totaling gross weights and dividing by the number weighed; enter preliminary average tare from caption 10 in block 11b; determine average net weight by subtracting block 11b from 11a; enter the declared net weight as stated on the package weighed; determine the shortage by subtracting block 11c from 11d. Block 12 Preliminary % Short - Enter the preliminary percent short, which is determined by dividing e by d. Block 13 Remarks - Record any observations on the condition of the lot or storage facilities which might affect net weights, (faulty machine sealing of packages, extreme high temperature, extended length of storage, etc.) Block 14 District - Enter the name of the collecting district. Block 15 Employee Signature - Sign the form. Block 16 Employee Title - Enter your title.
Field determination of volume is a screening procedure to determine the likelihood of short volume units in the lot. The laboratory will confirm both tare and net volume.
The approximate volume of small containers of free flowing liquids may be obtained by direct measurement. Standardized graduated cylinders calibrated to "contain" a given volume can be obtained from the laboratory. Use the smallest graduate that will hold the volume to be measured. Under no circumstances use a graduate to measure a volume less than 25% of the maximum capacity of the graduate. Proceed as follows:
- Select 8 units at random; one from each of 8 cases or otherwise representative of the lot.
- Empty contents into calibrated graduate holding the container in a nearly vertical position, but tipping so that the bottom of the container will drain. Allow to drain one minute after stream breaks into drops. Obtain an anti-foaming agent from the laboratory if beer or other product likely to foam are measured.
- Hold the graduate vertically with the surface of the liquid level with the eye. Place a shade of some dark material immediately below the meniscus and read volume from the lowest point of the meniscus. A convenient device for this purpose is a collar-shaped section of thick black rubber tubing cut open at one side and of such size as to clasp the graduate firmly.
- If no units containing less than declared volume are found, no further determinations are required.
- If one or more units containing less than declared volume are found, measure 4 additional units selected as above.
- If the total of twelve determinations contains only one short volume unit, be guided by the significance of the average shortage as related to the individual program guideline.
- If the total of twelve determinations contains more than one short volume unit, an Official Sample of 48 units should be collected regardless of the average shortage figure.
Direct measurement of viscous liquids or large containers is not practical. Field weigh 48 units as specified in IOM 22.214.171.124.3.
See the document "Guide to Nutritional Labeling and Education Act (NLEA) Requirements" for guidance. See Office of Nutrition, Labeling, and Dietary Supplements (ONLDS) website and FDA.gov for the most up-to-date information regarding claims in labeling.
Also, see CPGM 7321.005 to determine enforcement priorities for food labeling violations, including those related to the Food Allergen Labeling and Consumer Protection Act (FALCPA).
Examination of many products may be conducted on the spot without fixed laboratory equipment. These examinations vary from simple visual observations for gross filth, such as rodent pellets in wheat, to the detection of odors of decomposition in seafood. Organoleptic examinations for regulatory purposes shall be made only by those individuals qualified by training or experience to conduct such examinations. If it is necessary to collect physical subsamples for organoleptic examination and they are collected from bulk, the subs must be packed in glass jars to prevent the product from picking up foreign odors.
When making filth examination by screening shelled peanuts, dried bean, peas and similar products, packed in large containers (i.e., 50-125 lb. bags) use the portable folding whole-bag screens available in your district. Conduct the examination in a well-lighted area. Set up screen and adjust height to permit opening the bags directly onto the high side of the screen. Place another bag or container on the screen's low side to catch the screened product. Place a sheet of clean butcher or similar paper in screen body to catch screenings and insert screen wire over paper. Open stitches of bag being examined to permit approximately ten to twenty pound portions to enter onto high side of screen. Gradually work the product across the sieve to the low side and into the receiving container. Do not push large quantities rapidly across screen because insects, eggs, stones, excreta pellets, etc., will be carried along with the product and will not sift through the sieve openings. Examine the screening from each bag and subjectively report live or dead insects, rodent excreta pellets, or other obvious filth. Submit screenings as separate subs if actionable.