Nationwide Survey of Distillers Grains for Aflatoxins

Date: November 21, 2006

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

Subject: Nationwide Survey of Distillers Grains for Aflatoxins

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

ORA Concurrence # 2006100302
FACTS Assignment # 787470

Objectives:

1. To determine levels of aflatoxin, a toxicologically significant contaminant of corn and other grains/oilseeds, in a limited nationwide survey of distillers grains samples.
2. To prevent distillers grains that contain unacceptable levels of aflatoxins from entering interstate commerce for use in animal feeds and to initiate procedures to remove distillers grains that contain unacceptable levels of aflatoxins from interstate commerce.

Background:

FDA’s Center for Veterinary Medicine (CVM) is concerned about the potential animal and human health hazards from the use of aflatoxin-contaminated distillers grains as ingredients in animal feeds. Distillers grains are animal feed ingredients that are by-products of alcohol (spirits) and ethanol production . Approximately 90% of the distillers grains produced in U.S. facilities are used in domestic animal feed.

Since 1999, the production of ethanol has increased by more than 150 percent. In 1999, the annual U.S. ethanol production was 1.47 billon gallons. In 2005, the annual U.S. ethanol production had grown to 3.9 billion gallons. Current U.S. ethanol industry projections are that domestic ethanol production will exceed 7.5 billion gallons per year by 2012 . As a result of this increased ethanol production, the quantity of distillers grains marketed for use in animal feed has increased from 1.89 million metric tons in 1999 to 8.35 million metric tons in 2005 (a 340 percent increase), and is expected to continue to increase in the future.

In 1999, 0.72 million metric tons of distillers grains were fed to dairy cattle, but in 2004 that amount had grown to 2.8 million metric tons and this represented approximately 43% of all distillers grains used in animal feed. Increased use of corn and grain sorghum (milo) for ethanol production may lead to increased demand for these agricultural products, which in turn may lead to the increased use of corn and grain sorghum contaminated with aflatoxins in ethanol production. Researchers have shown that aflatoxins are not destroyed during ethanol production, but are concentrated in the distillers grains by-products where they may be present at levels as high as three times that found in the corn or grain sorghum starting materials. Thus, distillers grains contaminated with aflatoxins can pose risks to the safety of animals consuming these products and with the widespread use of distillers grains in dairy cattle feed there is also a potential human safety concern due to aflatoxin residues in the milk.

Currently, we have only limited data on the prevalence and levels of aflatoxins in distillers grains that are produced by U.S. ethanol facilities and marketed as animal feed ingredients. Since 1999, only 12 domestic samples of distillers grains have been collected and analyzed for aflatoxins by the FDA, and none of these samples had detectable levels of aflatoxins. Because of this limited data and because of the marked increase in the amount of distillers grains used in animal feeds since 1999, CVM believes it is important for the agency to conduct a limited nationwide survey at this time to determine the prevalence and levels of aflatoxin in distillers grains produced by U.S. ethanol production facilities. The agency wants to prevent distillers grains that contain unacceptable levels of aflatoxins from entering interstate commerce for use in animal feeds and plans to initiate appropriate administrative and/or regulatory action to remove distillers grains that contain unacceptable levels of aflatoxins from interstate commerce.

Assignment:

1. Overview of Samples to Collect

There are currently more than 100 ethanol facilities that manufacture distillers grains. Most of the ethanol production facilities are located in districts covered by the FDA Central and Southwest Regional Offices. The distillers grains produced by these plants may be sold directly to livestock producers, commodity brokers, and/or feed manufacturing facilities.

For the purposes of this assignment, distillers grains are feed ingredients that meet the Association of American Feed Control Officials Incorporated (AAFCO) definitions for distillers dried grains (DDG), distillers dried grains with solubles, or distillers wet grains. The definition numbers for these AAFCO feed ingredients are 27.5, 27.6, and 27.8 and are found on pages 273 and 274 of the AAFCO 2006 Official Publication. If you do not have, or cannot locate, a copy of the AAFCO Official Publication, please contact Dr. Michael Henry at CVM for assistance in obtaining copies of these definitions (phone: 240-453-6861; e-mail: Mike.Henry@fda.hhs.gov).

Of the 250 samples allocated to PAC code 71003C – Feed Contaminants Program Mycotoxins/Aflatoxins during fiscal year 2007, 40 samples should be used for this survey. The samples should be collected between November 2006 and September 30, 2007, from distillers grains manufactured at ethanol production facilities and offered for sale or use in animal feeds. The 40 distillers grains samples to be collected during this assignment are divided among 19 FDA district offices as described below.

DAL, DEN & KAN – 4 domestic samples from each district
BLT, CHI, CIN, DET, MIN & PHI – 3 domestic samples from each district
ATL, FLA, NOL, SJN, NWE, NWJ, NYK, SAN, SEA & LOS – 1 domestic sample from each district

CVM recommends that samples of distillers grains be collected for this assignment at ethanol production facilities or at the feed manufacturing facilities. CVM does not anticipate that districts will have any problems in locating ethanol production facilities that produce distillers grains for interstate sale as feed ingredients, or in locating feed manufacturers that use distillers grains in feeds. However, districts that have difficulty identifying ethanol production facilities that produce distillers grains or feed manufacturers that use distillers grains should contact Dr. Michael H. Henry at CVM for assistance (phone: 240-453-6861; e-mail: Mike.Henry@fda.hhs.gov).

Because corn and grain sorghum are harvested in the fall and stored for use throughout the year, CVM encourages districts to collect samples of distillers grains throughout the year, to the extent possible -- e.g., districts collecting more than one sample should try to collect half of the samples during the fall/winter seasons (November 2006 to March 2007) and half during the spring/summer seasons (April 2007 to September 2007).

2. Sample Collection, Sample Handling & Shipment

For collection of samples for mycotoxin analysis, refer to IOM Chapter 4 for sampling techniques, handling, shipping, and storage. Collect 10 sub-samples of distillers grains, each weighing ~454 grams (1 lb), from each sampled feed as shown in Sample Schedule 6 of IOM Chapter 4 for corn-shelled, meal or grit. Each of these 10 sub-samples already includes the 702(b) portions so there is no need to collect additional 702(b) portions.

During the collection, the investigator should also identify the source material (corn or grain sorghum) for the distillers grains and the intended use of the product.

Send the official sample to your FDA servicing laboratory, and report the time spent on its collection and submission under PAC 71003C – Feed Contaminants Program Mycotoxins/Aflatoxins.

Servicing labs:

Pacific, Northeast & Central Regions to PRL-NW
Southeast Region to SRL
Southwest Region to ARL

Refer to the Feed Contaminants Compliance Program and the FDA Investigations Operations Manual for more detailed information about sample collection, sample handling and shipment.

3. Sample Analysis

There are 34 rapid test kits that have been performance-verified by GIPSA (Grain Inspection, Packers and Stockyards Administration of the USDA) for analysis of mycotoxins. Sixteen of these 34 rapid test kits are used for measuring aflatoxins (8 quantitative and 8 qualitative). Distillers dried grains (DDG) and/or DDG with solubles are listed in the identified commodities column in 4 of the 8 rapid test kits used for quantitating aflatoxins ( http://archive.gipsa.usda.gov/tech-servsup/metheqp/testkits.pdf ). Any of these 4 rapid test kits can be used for analyzing samples of distillers grains for aflatoxins in this survey. Brief summaries of these 4 rapid test kits are provided below in Table 1.

Table 1. Rapid test kits that have been performance-verified by GIPSA for quantitating aflatoxins in DDG and/or DDG with solubles.

Company	Test Kit	Test Kit Range	Test Format	Identified Commodities
Charm Sciences	ROSA Aflatoxin (Quantitative)	5-100 ppb	lateral flow strip	distillers dried grains (DDG)
Neogen Corp.	Veratox	5-50 ppb	microtiter well plate	DDG with solubles
R-BioPharm	Ridascreen Fast SC	5-100 ppb	microtiter well plate	DDG with solubles
Vicam	Aflatest	5-100 ppb	immunoaffinity column	DDG and DDG with solubles

If a distillers grains sample contains more than 20 parts per billion (ppb) of total aflatoxins (aflatoxin B 1 + B 2 + G 1 + G 2) when analyzed by a rapid test kit, this result should be confirmed using a second method which can be selected by the servicing laboratory. The HPLC Reference Method for the Determination of Aflatoxins in Corn, Distillers Dried Grains and Corn Gluten Feed Samples would be an acceptable confirmatory method for the rapid test kit results. This HPLC reference method was developed by the UDSA/GIPSA/Technical Services Division/Analytical, Reference and Testing Services Branch. The USDA HPLC Reference Method has been sent to the Director of the three mycotoxin-servicing labs and the Division of Field Science (DFS).

There could be a wide range of moisture content in the distillers grains samples depending on whether they have been dried and the drying process used. Thus, the moisture content should be determined for each sample collected as part of this survey. The moisture content of distillers grains should be determined by AOAC Official Method 934.01, Loss on Drying (Moisture) at 95-100°C for Feeds (Official Methods of Analysis of AOAC International, 17 th edition, 2000, volume 1, 4.1.03).

FDA labs are encouraged, but are not required, to analyze the distillers grains for other mycotoxins -- fumonisins, vomitoxin, zearalenone and/or ochratoxin A. Please refer to the Feed Contaminants Program for information about which other mycotoxins are appropriate to test for in corn products, sorghum (milo) products, and other grain products (see analytical methods [Section IV], levels/classification [Section V] and priorities [Attachment A]).

In addition, resources are available this year under PAC 71003B – Feed Contaminants Program Elements for analysis of a limited number of animal feeds and feed ingredients. Since sulfuric acid is commonly added during the manufacturing process for distillers grains, FDA labs are encouraged, but not required, to analyze some of the distillers grains samples for sulfur and/or other elements under PAC 71003B.

All analytical results from these samples, including those for aflatoxins, other mycotoxins, elements, and the moisture content, should be reported in FACTS.

4. Primary Contact:

Please contact Dr. Michael H. Henry (Mike.Henry@fda.hhs.gov: 240-453-6861) if you have any questions about sample collection, sample handling, shipment and sample analysis.

Regulatory/Administrative Follow-up:

The FDA has established action levels for aflatoxins in animal feeds in FDA Compliance Policy Guide (CPG) Sec. 683.100. The action level for aflatoxins in feeds and feed ingredients for dairy cattle, immature animals, and animal species not specified in this CPG, or when the intended use is not known, is 20 parts per billion (ppb). The action level for aflatoxins in corn products, including corn distillers grains, intended for finishing (i.e., feedlot) beef cattle is 300 ppb. The action level for aflatoxins for corn products, including distillers grains made from corn, intended for finishing swine of 100 pounds or greater is 200 ppb. The action level for aflatoxins in corn products, including distillers grains made from corn, intended for breeding beef cattle, breeding swine or mature poultry is 100 ppb.

CVM intends to support administrative and/or regulatory actions (including, but not limited to, untitled letters, warning letters, recalls, seizures and injunctions) when aflatoxin levels in distillers grains that are intended for use in animal feeds and are in, or are likely to enter, interstate commerce exceed the action levels in the FDA CPG Sec. 683.100. Thus, districts with samples of distillers grains that exceed the aflatoxin action levels should submit a recommendation for administrative and/or regulatory action with supporting documentation to CVM for review.

Priority/Timeframe/Resources:

This assignment has a routine priority and the concurrence of ORA. The ORA concurrence # is 2006100302. This assignment is directly associated with the Feed Contaminants Program and resources have been previously allocated. Please report time for collection and analysis of samples under PAC 71003C – Feed Contaminants Program Mycotoxins/Aflatoxins. The anticipated sample collection and analytical times for each sample are about four and eight hours, respectively.

Contacts: CVM	Contacts: ORA
Scientific and Technical Questions: Dr. Michael H. Henry FDA/CVM/OSC/DAF/HFV-222 Phone: 240-453-6861 E-mail: mailto:Mike.Henry@fda.gov Compliance: Mr. Jack Geltman FDA/CVM/OSC/DC/HFV-232 Phone: 240-276-9203 E-mail: Jack.Geltman@fda.hhs.gov	Scientific and Technical Questions: Lawrence (Larry) D’Hoostelaere, Ph.D. FDA/ORA/ORO/DFS/SCRRB/HFC-141 Phone: 301-827-1032 E-mail: Lawrence.DHoostelaere@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

Please refer to FACTS assignment # 787470 when referring to this assignment.

Dan McChesney, Ph.D.
Director, Office of Surveillance & Compliance
HFV-200

cc:

Regional Food and Drug Directors
HFC-102 Lynch
HFC-132 Dunnie
HFC-141 D’Hoostelaere
HFC-200 McChesney, Graber
HFV-220 Benz
HFV-222 Henry, Lovell, Ekelman
HFV-226 Gill
HFV-230 Young, Geltman