Contents

4.3 - COLLECTION TECHNIQUE

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.

4.3.1 - Responsibility

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 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.

4.3.2 - LOT RESTORATION & IDENTIFICATION

4.3.2.1 - Restoring Lot(s) Sampled

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 diary 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 4.2.8.4.

4.3.2.2 - Identifying Lot(s) Sampled

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 diary.

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 4.5.2.3 for identification techniques.

Do not permanently identify articles that are borrowed and will be returned to the dealer.

4.3.3 - SAMPLE SIZE

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 4.3.3.2 and 4.3.3.3.

4.3.3.1 - Medical Device Samples

The following table represents the devices for which there are sampling instructions in Compliance Policy Guides:

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 should be recommended.

4.3.3.2 - 702(b) Requirement

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):

1. Devices are not included in the statutory requirement of Section 702(b).
2. 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.
3. 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.
4. Import samples, collected from a shipment being imported or offered for entry into the United States.
5. 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.
6. 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, a good rule of thumb to follow for most filth samples, is to collect the 702(b) portion.

4.3.3.3 - Collecting the 702(b) Portion

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 4.3.7.4.

4.3.4 - IN-TRANSIT SAMPLES

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.

4.3.4.1 - Examination without a Warrant

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:

1. The consignor or consignee affirmatively consents to examination and/or sampling of the contents; or
2. 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
3. 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.]

4.3.4.2 - Examination with a Warrant

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 Division of Field Investigations (DFI) (HFC-130) at 301-827-5653 to discuss the matter. They will coordinate as necessary with Office of Enforcement (HFC-200) and Chief Counsel (GCF-1) 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 1 or 2 above 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.

4.3.4.3 - Resealing Conveyances

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.

4.3.5 - SPECIAL SAMPLING SITUATIONS

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 these areas of the IOM:
IOM 1.5. - Safety
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 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 gasses - Gasses represent a special sampling situation. Please contact your servicing lab to determine an appropriate sampling container and sample size.

4.3.5.1 - Complaints, Counterfeiting / Tampering, Foodborne Disease, Injury Illness

Detailed instructions for investigating and sampling products in connection with consumer complaints, tampering, foodborne outbreaks, injury and adverse reactions, etc. appear in the following 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 8.8.5.3 - 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.

4.3.5.2 - Recalls

See IOM 7.1 and IOM 7.1.1.7

4.3.5.3 - Natural Disasters

See IOM 8.5.

4.3.5.4 - Induced Samples

If this type sample is desired your supervisor will provide specific instructions and procedures to be followed. This may involve:

1. 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.
2. 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.
3. Whether to use order blanks contained in the promotional package, advertisement, or promotional activity; or whether false ones will be used.
4. Whether money orders, your charge plate 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.
5. Where the requested items are to be sent: rented P.O. Box, home address, General Delivery, or other address.
6. 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.

4.3.5.5 - Undercover Buy

See IOM 4.1.4.6.

4.3.6 - ASEPTIC SAMPLE

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 are 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.

4.3.6.1 - General Procedures

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.

4.3.6.1.1 - Sterilized Equipment

Use only sterilized equipment and containers. These should be obtained from the servicing laboratory or in 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.

4.3.6.1.2 - Cautions

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 4.3.6.5.

4.3.6.1.3 - Opening Sterile Sampling Containers

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. Submit one empty sterile container similarly opened and closed as a control. See IOM 4.3.6.5.

4.3.6.1.4 - Dusty Areas

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. 

4.3.6.2 - Sampling Dried Powders

Cautions - The proper aseptic sampling of dried milk powder, dried eggs, dried yeast, and similar type 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.

4.3.6.2.1 - Bag and Poly-liner Stitched Together Across Top Seam

1. 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.
2. 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.
3. Carefully scrape off the surface of the product with a sterile device and aseptically draw the sample from the material below.
4. Carefully reclose the bag and restitch by hand, or by machine if firm or FDA portable sewing machine is available.

4.3.6.2.2 - Bag Stitched Across Top and Poly-liner Twist-Closed and Sealed with "twist" Device - Wire, Plastic, etc.

1. Brush, alcohol wipe, and remove stitching as described.
2. Remove "twist" seal and carefully open poly-liner using caution that no extraneous material drops into the product.
3. Draw aseptic sample in same manner as in 3. above.
4. Carefully close the poly-liner with a twisting motion and reseal with "twist" seal arranging it so it will not puncture the poly-liner, and resew bag as in 4. above.

4.3.6.2.3 - Bags with Filling Spouts

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.

1. 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.
2. 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.
3. Carefully close the spout with a firm twisting motion and be sure the opening is closed prior to pushing back into the bag.

4.3.6.3 - Collecting Water Samples

When it is necessary to collect water samples for bacteriological examination, use the following procedures:

1. 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.
2. 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.
3. Clean and dry outside of faucet.
4. 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.
5. Partially close faucet to permit collecting sample without splashing. Carefully open sample bottle to prevent contamination, as for any other aseptic sampling operation.
6. 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.
7. Unless otherwise instructed, minimum sample size for bacteriological examination is 100 ml.
8. 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.

4.3.6.4 - Sample Handling

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 4.5.3.5 and 8.3.3.3. 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^oC (100^oF).

Submit samples subject to rapid spoilage (specimens of foods involved in poisoning cases, etc.) by immediate personal delivery to the bacteriologist where feasible.

4.3.6.5 - Controls

When collecting samples using aseptic technique and the subs are collected using presterilized containers and equipment, submit a number of control subs. If the sampling covers a long period of time you should submit controls which show environmental conditions during the time of sampling. The controls should be collected at the start, during, and at the end of the sampling period. List control subs on your C/R.

Examples of various control subs are:

1. 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. Also submit at least one empty sterile container which has been opened and closed in the sampling area.
2. Sterile Disposable Gloves - If sterile disposable gloves are used to handle the product, submit one unopened pair of gloves as a control.
3. 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.

4.3.7 - ADULTERATION VIOLATIONS

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 4.3.7.3.  You will need to field examine (See IOM 4.3.7.1) a number of lots of product to determine the extent of the adulteration and can collect an investigational (INV) sample of filth exhibits and take photographs to document the widespread nature of the evidence.  See IOM 4.1.7.  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.

4.3.7.1 - Field Examination

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.

4.3.7.2 - Random Sampling

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 4.3.3.2.

4.3.7.3 - Selective Sampling

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.

4.3.7.4 - Sample Criteria

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 4.3.7.2 - 4.3.7.4.  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:

1. Current or old
2. Isolated to one lot (possible FD&C 402(a)(3) charges - contain in whole or in part  filth or is otherwise unfit for food).
3. 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 you observe by 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.

4.3.7.4.1 - General

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.

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:

1. Use a coherent numbering/identification system for subsamples to avoid unnecessary confusion for the lab.
2. Provide a detailed listing of individual sub descriptions on the C/R.
3. If possible, provide a copy of any maps, photos or other additional documentation to the laboratory.
4. 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.
5. 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.

4.3.7.4.2 - Rodent Contamination

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:

1. Three or more of the bags in the lot are rodent gnawed; or
2. 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
3. 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:

1. 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:
2. 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:
3. 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.

4.3.7.4.2.1 - Examination and Documentation of Rodent Contamination

Note clearly on your C/R if the product or package contains or is directly associated with any of the following:

1. Dried milk products (contain urea).
2. Whole grain wheat (contains urea and allantoin).
3. Animal feeds (urea is usually intentionally added).

4.3.7.4.2.2 - Collecting Exhibits or Subsamples

When sampling lots for rodent contamination follow the safety precautions in IOM 1.5.5.4. Wear gloves and handle the exhibits with tweezers or forceps.

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 4.3.7.4.1.

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 702b subsamples as such on subsample identification  (See IOM 4.5.2.1.) and note on your C/R which subsamples are the 702(b) portions.

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.

  Collect nesting material with minimal handling. A half cup is enough for analysis.  Do not collect any live rodents.

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.

Handle exhibits carefully to prevent loss of microscopic evidence.

Submit each portion of bagging or container portion, 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.

4.3.7.4.2.3 - Summary of Sample For Rodent Evidence

The complete official sample will consist of:

1. Subsamples of rodent excreta pellets
2. Subsamples of stained bagging, or portions of the containers, and any adhering pellets.
3. Subsamples of unstained bagging, or portions of the containers, which do not fluoresce, for controls (minimum three required).
4. 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.
5. Subsamples of small portions of product to serve as 702(b) portions
6. 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
7. Subsamples of small amounts of product collected from beneath the gnawed holes.
8. Subsamples of  small portions of product to serve as 702(b) portions. 
9. Product labeling.
10. Interstate documentation.

4.3.7.4.3 - Insect Contamination

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.

1. The product contains:
   1. 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
   2. 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.
2. The product contains one or more live insects in each of three or more immediate containers.
3. 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.
4. 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.

4.3.7.4.3.1 - Examination and Documentation of Insect Contamination

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.

4.3.7.4.3.2 - Collecting Exhibits or Subsamples

Collect a representative number of insects for laboratory confirmation. 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.

4.3.7.4.3.3 - Summary of sample for insect evidence

The complete official sample will consist of:

1. Subsamples of insects, larvae, webbing, etc.
2. Subsamples of portions of the containers with entrance or exit holes.
3. Subsamples of small portions of the product from directly beneath holes.
4. Subsamples of small portions of product serve as 702(b) portions  See IOM 4.3.7.4.1.
5. Product labeling.
6. Interstate documentation.

4.3.7.4.4 - Bird Contamination

Per the criteria from CPG 580.100, if the product is in permeable containers (paper, cloth, burlap, etc.), and

1. The product contains bird excreta in one or more containers, and you feel the insanitarystorage conditions will clearly support a  402(a)(4) [21 U.S.C. 342 (a)(4)] violation.
2. Bird excreta is present on the exteriors of at least five of the containers, and the product contains bird excreta in one.
3. 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.

4.3.7.4.4.1 - Examination and Documentation of Bird Contamination

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.

4.3.7.4.4.2 - Collecting Exhibits and Subsamples

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 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, 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 4.3.7.4.1).

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 702b subsamples, as such on subsample identification  (See IOM 4.5.2.1.)  Note on the subsample itself and on your C/R which subsamples are the 702(b) portions.

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.

4.3.7.4.4.3 - Summary of Sample for Bird Evidence

The complete official sample will consist of:

1. Subsamples of stained bagging, or portions of the containers.
2. Subsamples of unstained bagging, or portions of the containers  for controls (minimum three required).
3. 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.
4. Subsamples of  small portions of product to serve as 702(b) portions.
5. 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.
6. Product labeling.
7. Interstate documentation.

4.3.7.4.5 - Chemical Contamination

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.

4.3.7.4.6 - Mold Contamination

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:

1. A 10 lb. sample representative of the corn lot must be obtained by probing, or by continuously sampling a grain stream.
2. Examine using a 366 nm UV light (portable black-lights meet this criteria).
3. 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.
4. Use a 2 lb. Portion, and carefully observe the entire corn surface one kernel at a time. Examine the entire sample using this procedure.
5. 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.
6. If four (4) or more BGYF particles are detected in the 10 lb screening sample, collect a sample for laboratory analysis.

4.3.7.5 - Abnormal Containers

 See IOM SAMPLE SCHEDULE CHART 2 - LACF for listing can defects.

4.3.7.6 - In-Line Samples

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% formalin solution. Large amounts of slime may be placed in a wide mouth glass jar with either a 1% formaldehyde solution or a 3% 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 furnish the 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 (vinegar).

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.

Bacteriological Samples - During inspections of firms producing products susceptible to microbial contamination (e.g., frozen precooked; ready to eat seafood, creme filled goods, breaded items, egg rolls, prepared salads, etc.), proof of adulteration, with fecal organisms, or elevated levels of non-pathogenic microorganisms, must be established. Sampling of raw materials, in-line and finished product is warranted. Follow instructions under IOM 4.3.7.7 - Products Susceptible to Contamination with Pathogenic Microorganisms, Sampling During Inspection.

4.3.7.7 - Products Susceptible to Contamination with Pathogenic Microorganisms

A top priority of the agency and CFSAN is to decrease foodborne illness caused by microbial contamination. With the rise of foodborne outbreaks detailed guidance was developed for sample collections and inspections dealing with microbial contamination.
Note: This guidance is intended to augment guidance found in the Compliance Programs listed below. Instructions in current compliance programs and ongoing assignments supersede this guidance. Before conducting inspections under these programs, investigator/analyst teams should be thoroughly familiar with the guidance provided in the appropriate Compliance Program.

For the following Compliance Programs collect samples for microbiological analyses only if:

1. Directed to do so in the current compliance program or ongoing assignments.
2. The firm has a previous history of microbiological contamination (e.g., follow up to illness or injury complaint, recalled/seized product, previous inspectional history, etc.) or
3. Sampling is conducted 'for cause' during an inspection (e.g., inspectional observations warrants collection for microbiological analyses):
   1. Domestic and Imported Cheese and Cheese Products (7303.037)
   2. Domestic Food Safety (7303.803)
   3. Domestic Acidified and Low-Acid Canned Foods (7303.803A)
   4. Domestic Fish and Fishery Products (7303.842) and
   5. Juice HACCP Inspection Program (7303.847).  Except as directed by the compliance program, you should not conduct any in-line, environmental, or finished product sampling, for microbiological concerns, during the inspection. Instead, fully document the lack of HACCP control(s) without physical sampling. If in the investigator's judgment the firm's HACCP plan is extremely inadequate and therefore sampling is warranted, contact CFSAN/OC/Division of Enforcement/Domestic Branch HFS-607 to determine what in-line sampling will be performed and the type of regulatory action that may be warranted for the situation at hand.