Nationwide Assignment to Collect Samples of Direct-Human-Contact Feeds and Analyze Them for Salmonella and E. coli O157:H7

Date: October 17, 2006

From: Director, Office of Surveillance and Compliance (HFV-200)

Subject: Nationwide Assignment to Collect Samples of Direct-Human-Contact Feeds and Analyze Them for Salmonella and E. coli O157:H7

To: All FDA District Directors, FDA Regional and District Laboratory Directors, and FDA District Investigations Branch Directors

ORA Concurrence # 2007091101
FACTS Assignment Code #777588

Objectives:

1. To determine the prevalence of Salmonella and E. coli O157:H7 in a limited number of direct-human-contact feeds nationwide.
2. To determine the serotype, genetic fingerprint, and/or antimicrobial susceptibilities of each Salmonella and E. coli O157:H7 isolated from direct-human-contact feeds collected under this assignment.
3. To remove direct-human-contact feeds found to be contaminated with Salmonella and/or E. coli O157:H7 from commerce.

Background:

FDA’s Center for Veterinary Medicine (CVM) is concerned about the potential for animal feeds to serve as vehicles for transmitting pathogenic and antibiotic-resistant bacteria to humans and other animals, and is particularly concerned about the potential for direct-human-contact feeds to directly transmit Salmonella and E. coli O157:H7 to humans.

Direct-human-contact feeds are animal feeds that are intended for use in feeding animals in homes, pet stores, petting zoos, zoos, fairs, or festivals. The feeds are likely to be directly handled or ingested by individuals, including certain vulnerable populations such as children, the elderly, and persons with compromised immune systems. Direct-human-contact feeds include, but are not limited to:

• Pet treats, chews, or snacks
• Pet foods
• Vitamins, minerals, or mixtures of vitamins, minerals, and other feed ingredients. These products are manufactured in various forms (pills, boluses, solutions, powders, etc) and intended for dogs, cats, and other pets
• Feed ingredients or mixtures of feed ingredients intended to be given as food or treat to animals in pet stores, petting zoos, zoos, fairs, or festivals.

There are numerous reports of human salmonellosis or E. coli O157:H7 infections in the United States (US) in which the illnesses were linked to human contact with various pets at homes, pet stores, petting zoos, zoos, fairs, and festivals. Pets implicated in these cases or outbreaks include chicks, ducklings, rabbits, livestock (cattle, sheep, and goats), and reptiles (bearded dragons, iguanas, lizards, snakes, and turtles).

In most of these reports, the authors did not determine if feeds fed to the animals were implicated in the outbreaks/cases or played a role in the illnesses. However, in one of the reports on pet turtle-associated human salmonellosis, the authors found the causative Salmonella in the turtle’s drinking water. Also, in one of the reports in which outbreaks of human E. coli O157:H7 infections were associated with visits to petting zoos, the authors state that 32 % of affected individuals had fed livestock at a petting zoo. In addition, there are published reports of three recent outbreaks of human salmonellosis in Canada and the US that were linked to Salmonella contained in pig ears and other pet treats fed to dogs. These three outbreaks occurred between 1999 and 2005, and two of the outbreaks were associated with pet treats manufactured in Texas and Washington.

For these reasons, CVM considers it prudent to keep Salmonella or E. coli O157:H7-contaminated direct-human-contact feeds out of interstate commerce.

Assignment:

1. Overview of Samples to Collect

Resources have been allocated under the fiscal year (FY) 2007 workplan to collect and analyze 200 official samples of feed under PAC code 71003E and to collect 200 investigational samples of feed under PAC code 71003E. All of the 200 official and 200 investigational samples in the workplan will be collected (both the 200 official feed samples and 200 investigational feed samples) and/or analyzed (the 200 official feed samples only) under this assignment.

All time spent collecting and analyzing the 200 official samples should be reported under PAC code 71003E. To be consistent with the Feed Contaminants Program ().

Due to reports of seasonal variations in the prevalence of Salmonella in feeds, districts are encouraged to spread out their sample collection throughout the fiscal year e. g., collect one quarter of your assigned samples during the fall (October, November, and December 2006), one quarter during the winter (January, February, and March 2007), one quarter during the spring (April, May, and June 2007), and one quarter during the summer (July, August, and September 2007). If this is not feasible, districts should try to collect half of the assigned samples during the spring and the other half during the summer. If a district is only able to collect assigned samples during one season of the year, we recommend that samples be collected during summer.

2. Sample Collection

For both official and investigational samples, follow the procedure for aseptic sample collection as described in the FDA Investigators Operations Manual (IOM) Subchapter 4.3 Collection Techniques. For official samples, ensure that the proper controls are collected as outlined in IOM 4.3.6.5 and that interstate commerce is documented.

During the surveillance portion of this assignment, each district should aseptically collect two official and two investigational samples from each of the four categories of direct-human-contact feeds identified in the four bulleted paragraphs below:

• Pet treats, chews, or snacks. Examples include, but are not limited to, rawhide bones, pig ears, dog biscuits, salmon treats, and shrimp snacks.
• Non-canned pet foods. Examples include, but are not limited to, non-canned foods for puppies, dogs, kittens, cats, rabbits, reptiles (e. g., turtles, snakes, lizards, and iguanas), rodents (e. g., gerbils, hamsters, guinea pigs, rats, and mice), birds, and aquarium fish.
• Vitamins, minerals, or mixtures of vitamins, minerals, and other feed ingredients in the form of pills, boluses, solutions, or powders intended for dogs, cats, and other household pets.
• Feed ingredients, or mixtures of feed ingredients, intended to be given as food or treat to animals in pet stores, petting zoos, zoos, or at local/regional/state/national fairs and festivals. The preferred samples to collect in this category are pellets, crumbles, boluses, or seeds that are already dispensed into small bags for purchase and use by the public, or in coin-operated feed dispensers.

Use the following guidelines when collecting samples:

• For packaged pet treats, and other direct-human-contact feeds contained in packages weighing less than 0.91 kg (<2 lbs), randomly select 10 (for official samples) or 1 (for investigational samples) intact packages and place them into separate, appropriately sized, sterile Whirl-Pak® bags or sterile, capped plastic bottles. If each package weighs less than 200 g (7.1 oz), combine intact packages as needed to achieve the desired sub-sample weight.
• For packaged pet treats and other packaged direct-human-contact feeds weighing 0.91 – 4.54 kg (2 to 10 lbs), randomly select three or more intact packages. For official samples, collect up to 4 sub-samples from different sections of each package, for a total of 10 sub-samples. Each of the 10 sub-samples should weigh ~200 g (7.1 oz) and should be placed into a sterile Whirl-Pak® bag or sterile capped plastic bottle. For investigational samples, collect a ~300 g (10.6 oz) portion and place in an appropriately sized sterile Whirl-Pak® bag or sterile capped plastic bottle.
• For packaged pet treats and other packaged direct-human-contact feeds weighing over 4.54 kg (>10 lbs), randomly select two or more intact packages. For official samples, collect up to 5 sub-samples from different sections of each package, for a total of 10 sub-samples. Each of the 10 sub-samples should weigh ~200 g (7.1 oz) and should be placed into a sterile Whirl-Pak® bag or sterile capped plastic bottle. For investigational samples, collect a ~300 g (10.6 oz) portion and place in an appropriately sized sterile Whirl-Pak® bag or sterile capped plastic bottle.
• For direct-human-contact feeds available in bulk, randomly select two or more containers of the bulk feed. For official samples, collect up to 5 sub-samples from different sections of each container, for a total of 10 sub-samples. If all of the feed is in one bulk container, collect all 10 sub-samples from different sections of the container. Each of the 10 sub-samples should weigh ~200 g (7.1 oz) and should be placed into a sterile Whirl-Pak® bag or sterile capped plastic bottle. For investigational samples, collect a ~300 g (10.6 oz) portion and place in an appropriately sized sterile Whirl-Pak® bag or sterile capped plastic bottle.
• For feeds that are already dispensed into small bags for use by the public at zoos and petting farms, consider each bag (if 200 g or larger) or an appropriate combination of bags (if each bat is less than 200 g) as a sub-sample. Randomly select 10 sub-samples for each official sample or 1 sub-sample for each investigational sample. Place each sub-sample into a separate appropriately sized sterile Whirl-Pak® bag or sterile capped plastic bottle.
• For feeds that are already placed in coin-operated feed dispensers for use by the public at zoos and petting farms, consider each serving (if 200 g or larger) or combination of servings (if each is less than 200 g) as a sub-sample. Collect 10 sub-samples for each official sample or 1 sub-sample for each investigational sample. Place each sub-sample into a separate appropriately sized sterile Whirl-Pak® bag or sterile capped plastic bottle.

Official and investigational samples can be collected on the same day and from the same lot of feed, but should be assigned different FACTS sample numbers.

• Official samples: Each FDA district will aseptically collect a total of eight official samples (two from each of the four types of direct-human-contact feed) during the surveillance portion of the assignment, and aseptically collect additional official samples as needed during follow-up investigation(s) of surveillance samples that were found to be positive for Salmonella or E. coli O157:H7. Every official sample should consist of 10 sub-samples weighing ~200 g (7.1 oz) each. Each of these 10 sub-samples already includes the 702b portions so there is no need to collect additional 702b portions.
• Investigational sample: Each FDA district will aseptically collect a total of eight investigational samples (two from each of the four types of direct-human-contact feed) during the surveillance portion of the assignment, and aseptically collect additional investigational samples as needed during follow-up investigation(s) of official samples that were found to be positive for Salmonella or E. coli O157:H7. Each investigational sample should consist of one sub-sample weighing ~300 g (10.6 oz). There is no need to collect 702(b) portions and control samples, or to document interstate commerce, for the investigational samples because they will be used for research purposes only and are not considered to be official samples.

3. Sample Handling and Shipment

Please refer to IOM Subchapter 4.5 for details about sample preparation, handling and shipping. Samples collected under this assignment will in general not require refrigeration, and should be shipped by overnight mail to the laboratory as soon as possible after collection. In those occasional instances where refrigeration may be indicated (e. g., moist feed, raw meat diets, etc), investigators should refer to the IOM, or contact Dr. Ekperigin, for guidance on handling and shipment.

Official samples: Each official sample, consisting of ten sub-samples (~200 g or 7.1 oz per sub-sample), may be used for regulatory purposes and should be sent to the pertinent FDA servicing laboratory as follows:

Servicing labs:
Central Region	CHI & MIN to DEN-LAB; DET to ARL; BLT, CIN, NWJ & PHI to SRL
Southwest Region	DAL, DEN & KAN to DEN-LAB; SWID to ARL
Northeast Region	all to NRL
Pacific Region	all to SAN-LAB
Southeast Region	all to SRL

Report time used in collecting and submitting the official samples under PAC 71003E.

Investigational samples: Each investigational sample (~300 g or 10.6 oz) will be used for research purposes only, and will be sent to Dr. Terry Proescholdt at CVM Office of Research [MOD II, Room 1515, HFV-530, 8401 Muirkirk Road, Laurel, MD 20708; Phone: (301) 210-4023; E-mail address: Terry.Proescholdt@fda.hhs.gov or his designee and should be labeled “Investigational Sample for Research Purposes Only.”; The investigational sample should be identified with the FACTS sample number, the collector’s name, the date of collection, the state (Oklahoma, New Jersey, etc.) in which the sample was collected, and a description of the sample and its intended use. Report time used in collecting and submitting the investigational samples under PAC 71R800.

4. Sample Analysis

Official samples: Analyze each official sample of direct-human-contact feeds for Salmonella and E. coli O157:H7. Use the methods described in the Bacteriological Analytical Manual (BAM) Online (see chapter 5 for Salmonella and chapter 4A for E. coli O157:H7).

For Salmonella, analyze a 375 g (13.2 oz) composite consisting of about 37.5 g (1.3 oz) from each of the 10 sub-samples. Prepare pet treats, chews and snacks, for Salmonella analysis using the procedures outlined in BAM, chapter 5, section C (Preparation of foods for isolation of Salmonella), sub-section 20 (Pig ears and other types of dog chew pieces).

For E. coli O157:H7, analyze each sub-sample individually using 25 grams (0.9 oz) from each sub-sample. Be certain to follow the procedures (enrichment, isolation, media preparation, presumptive screening, confirmation, etc.) in Chapter 4A of BAM that are specific for E. coli O157:H7.

Reserve approximately 75 grams (2.6 oz) or more from each of the 10 sub-samples for use as the 702(b) portion.

For samples that are found positive for Salmonella, the servicing lab will send each isolate to ARL or DEN-LAB for serotyping (NRL and SRL to ARL; SAN-LAB to DEN-LAB). ARL will send serotyped Salmonella isolates to DEN-LAB. DEN-LAB will conduct antimicrobial susceptibility testing on every Salmonella isolate found in animal feed.

For samples that are found positive for E. coli O157:H7, the servicing lab will send each isolate to DEN-LAB for antimicrobial susceptibility testing.

In addition, the servicing lab will determine the pulsed-field gel electrophoresis (PFGE) patterns of each Salmonella and E. coli O157:H7 isolate found in animal feed. If feasible, conduct the PFGE analysis on animal feed isolates separate from human food isolates. The servicing lab will send the image(s) of the PFGE patterns from the animal feed isolates to Mr. Jason Abbott at CVM Office of Research (CVM-OR) [MOD II, Room 1501, HFV-530, 8401 Muirkirk Road, Laurel, MD 20708; Phone: (301) 210-4185; E-mail Address: Jason.Abbott@fda.hhs.gov] or his designee. Include the FACTS sample number when electronically sending each PFGE image to CVM-OR. CVM-OR will be responsible for sending the PFGE image(s) from the animal feed isolates to PulseNet.

Investigational samples: CVM-OR will not analyze any of the investigational samples collected under PAC 71R800 in this assignment for Salmonella or E. coli O157:H7.

5. Primary Contact

Please contact Dr. Henry Ekperigin (Henry.Ekperigin@FDA.HHS.GOV: 240-453-6868) if you have any questions about sample collection, sample handling and shipment, and sample analysis.

Regulatory/Administrative Follow-up:

The Center intends to support regulatory action to keep Salmonella or E. coli O157:H7-contaminated direct-human-contact feeds found under this assignment out of interstate commerce.

Districts that find Salmonella or E. coli O157:H7 in the samples of direct-human-contact feeds collected should recommend the issuance of a Warning Letter for each positive sample and initiate procedures to remove the contaminated feed from interstate commerce. In 90 days or less after the occurrence of the violation, districts should conduct follow-up investigations of the facility or facilities where the violative sample was manufactured to review manufacturing and handling procedures, document what corrections, if any, were made and collect follow-up samples.

Priority/Timeframe/Resources:

This assignment has a routine priority and the concurrence of ORA. The assignment is a part of the Feed Contaminants Compliance Program and resources have previously been allocated to it under the FY 2007 Work Plan. Report time for the collection, submission, preparation, and analysis of each official sample (10 sub-samples, each weighing ~200 g) under PAC 71003E. Report time for the collection and submission of each investigational sample (one ~300 g sample) under PAC 71R800. The anticipated collection time is about four hours for each sample.

Contacts: CVM	Contacts: ORA
Scientific and Technical Questions: Dr. Henry E. Ekperigin FDA/CVM/OSC/DAF/HFV-222 Phone: 240-453-6868 E-mail: Henry.Ekperigin@fda.hhs.gov	Scientific and Technical Questions: Ms. Marsha A. Hayden FDA/ORA/ORO/DFS/SCRRB/HFC-141 Phone: 301-827-1039 E-mail: Marsha.Hayden@fda.hhs.gov Ms. Lydia Rosas-Marty, Microbiologist FDA/ORA/ORO/DFS/SCRRB/HFC-141 Phone: 301-827-6624 E-mail: Lydia.Rosas-Marty@fda.hhs.gov
Compliance: Mr. Jack Geltman FDA/CVM/OSC/DC/HFV-232 Phone: 240-276-9203 E-mail: Jack.Geltman@fda.hhs.gov	Investigations: Mr. James L. Dunnie Jr. FDA/ORA/ORO/DFI/DB/HFC-132 Phone: 301-827-5652 E-mail: James.Dunnie@fda.hhs.gov

/signed/
Daniel G. McChesney, Ph.D.
Director, Office of Surveillance and Compliance

cc:

Regional Food and Drug Directors
HFC-102 Lynch
HFC-132 Dunnie
HFC-141 Hayden, Rosas-Marty
HFC-200 McChesney, Graber
HFV-220 Benz
HFV-222 Ekperigin, Lovell, Ekelman, Kaplan
HFV-226 Gill
HFV-230 Dunnavan, Young, Geltman
HFV-530 (Abbott, Proescholdt)