Aflatoxins in Corn: How To Sample Corn For Aflatoxin Testing

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Aflatoxins in Corn

flatoxins are a group of chemicals


produced by certain mold fungi. These
fungi, Aspergillus flavus and Aspergillus
parasiticus, can be recognized by yellow-green or
gray-green, respectively, on corn kernels, in the field
or in storage (Figure 1). Although aflatoxins are not
automatically produced whenever grain becomes
moldy, the risk of aflatoxin contamination is greater in
damaged, moldy corn than in corn with little mold.
Aflatoxins are harmful or fatal to livestock and are
considered carcinogenic (cancer-causing) to animals
and humans. In the Midwest, aflatoxin levels are highest
during hot, dry summers. The prime conditions for the
fungus to produce toxin are warm August nights in a
period of drought.
In high-risk years, aflatoxin screening may be done
at the elevator or where the corn is marketed. Rapid,
on-site tests can determine the possible presence of
aflatoxin, but they do not provide specific quantitative
results. The toxins are produced inside the corn kernels
and their presence can be determined only by specific
analytical tests. Because aflatoxin levels can vary greatly
from kernel to kernel, sampling the load, bin, or unit
of grain is the most critical step in determining actual
levels of aflatoxin.

How to sample corn for


aflatoxin testing
Because aflatoxin does not occur uniformly through
a lot of grain and is usually localized in a small area,
the best approach is to make a composite sample
consisting of subsamples from every part of a load,
bin, or unit of corn. The recommended procedure is to
sample periodically from a moving stream, combining
these samples to obtain a composite sample of at least
10 lb of corn. An alternative is to sample with a probe
through a storage unit (five perimeter samples and one
center sample for each 6 feet of bin height). In the field,
sample individual fields or parts of fields separately.
Grain in trucks can probed in the same way to collect
samples of at least 5 lb per truck.
Fields that vary in cropping history, tillage
practices, planting date, soil type, or hybrid can differ
greatly in aflatoxin vulnerability. Sample a minimum
of 10 to 30 locations within each field. To reach the
same sampling frequency as testing grain in trucks,
collect one sample (5-10 lb) for about every 5 acres.
Immediately dry samples to 1214 percent moisture to
prevent aflatoxin development during transit or storage.
High-moisture samples should be frozen and delivered
to the laboratory in the frozen state. Dried samples
maintain their quality best if shipped in cloth or paper
containers.

Figure 1. Aspergillus ear rot symptoms on corn ear (left) and growth of Aspergillus flavus in artificial
culture (right).

PM 1800 Reviewed May 2009

How to test for aflatoxin


Currently, two types of screening tests are
available: blacklight tests and commercial test kits.
The blacklight (also called ultraviolet light) test is a
visual inspection for the presence of a greenish gold
fluorescence under light at a wavelength of 365 nm
(nanometers). The greenish gold fluorescence looks
like a firefly glow. More than four glowing particles per
5-pound sample (before grinding) indicate a likelihood
of a +20 ppb (parts per billion) level of aflatoxins.
However, remember that this test is an initial screening
for the presence of aflatoxin and the results should be
verified by laboratory analysis. If there are less than
four glowing particles per 5 lb sample, this does not
guarantee that the sample is free of aflatoxins.
Commercial test kits with immunoassay or ELISA
techniques are available for on-site tests for aflatoxin.
Immunoassay analysis is based on the detection of
specific proteins found in aflatoxins using antibodies
to identify these proteins. The tests are very specific for
aflatoxin, but they require operator training and practice
to be accurate. Some tests determine only the presence
or absence of aflatoxin; others can quantify, within a
range, the amount of aflatoxin present. If a lot of corn
is rejected based on the results of an immunoassay test
kit, the results also should be confirmed by laboratory
analysis. The entire sample should be ground before
removing a subsample for the test kit.
Analytical laboratories use one of several
procedures such as thin-layer chromatography, mini
columns, gas chromatography, or mass spectroscopy
to determine aflatoxin levels. These procedures are
highly accurate and quantitative. The laboratory should
grind the entire sample of corn together before taking
subsamples for analysis.

Corn samples to be analyzed can be sent to the


Iowa State University (ISU) Plant Disease Clinic,
to official USDAFGIS (United States Department
of AgricultureFederal Grain Inspection Service)
laboratories, or to a private laboratory. The ISU
Veterinary Diagnostic Laboratory analyzes samples
submitted through a veterinarian. Contact your local
extension office or go to www.iowagrain.org for a
current list of public and private laboratories equipped
for aflatoxin analysis.

Regulations regarding aflatoxin


in corn
The Food and Drug Administration (FDA) has
established an action level of 20 ppb for aflatoxins
in corn in interstate commerce. This is the level at
which federal agencies may take action, including
seizure of the corn or prohibition of its sale. Elevators
do not accept corn with 20 ppb or more of aflatoxin
unless they have a known alternative use. Even one
contaminated kernel in a 5-lb sample could result in
more than 20 ppb aflatoxin. The FDA has guidelines
for using contaminated grain in livestock feed (Table 1).
These guidelines are based on maintaining performance
and avoiding disease related to aflatoxin, except for
dairy cattle where prevention of carcinogenic aflatoxin
residues in milk is the concern.

Consequences of high
concentrations of aflatoxin in corn

Aflatoxins are very potent compounds that cause a


variety of human and animal health problems. On rare
occasions, livestock can die from ingesting aflatoxincontaminated feed. Most commonly, aflatoxin reduces
the feed efficiency and reproductivity of livestock. It
can suppress the immune system of animals, leading
to more frequent occurrence of infectious diseases.
Unfortunately, the most abundant
aflatoxin, aflatoxin B1, is a
Table 1. FDA guidelines for acceptable aflatoxin level
carcinogen. This raises human
in corn based on intended use.
health concerns because aflatoxin
can appear in the milk of dairy
Intended use
Aflatoxin level (ppb)
cows fed contaminated corn.
Milk (Dairy Feed)
None detected
Corn of unknown destination
<20
Corn for young animals
<20
Corn for dairy cattle
<20
Corn for breeding beef
<100
cattle, swine, and mature poultry
Corn for finishing swine
<200
Corn for finishing cattle
<300

2 Aatoxins in Corn

How to prevent aflatoxin in corn


In Iowa, problems associated with Aspergillus and
aflatoxins are most common in hot, dry years. The fungi
survive in plant residues and produce abundant spores.
The spores are carried by the wind and infect silks or
kernels, usually through insect wounds. The fungi grow
best in weather that is hot and sunny, warm at night,
and dry during the silk and fill stage. Injury by insects,
hail, drought stress, and early frost expose the kernels
to infection. Insects can help spread the fungus within
infected ears.
The amount of aflatoxin produced in storage is
determined by storage conditions. The most important
factors are grain moisture content and temperature.
Optimum storage temperatures for A. flavus to grow
are 8090F; optimum grain moisture content is 18
percent. Damaged corn also favors the growth of A.
flavus. Importantly, aflatoxin concentration never
decreases in storage; it only increases or remains the
same.
The key to preventing ear rot and storage mold
problems is detecting them early, in the field and in
the bin. The following practices can reduce aflatoxin
production in grain:
1. Control insects in the field. Second-generation
European corn borers and corn earworms
damage the ears, allowing for infection.
2. Scout. Early detection can prevent serious
losses and avoid crises. Obtain good advance
information as to the potential in your area.
Scout at black-layer (physiological maturity)
and again a couple weeks before harvest.
Look at the primary ear for insect damage and
Aspergillus spores (Figure 2).

4. Clean bins and grain-handling equipment and


remove fines from the corn before storing.
Old corn residue is frequently a source of
contamination.
5. After the harvest, clean corn can be kept at
16 or 17 percent moisture during the winter.
Moldy corn should be dried immediately to
15 percent moisture or less. Holding grain for
even a short time can allow significant mold
and mycotoxin development. For long-term
storage, all corn should be dried to 14 percent,
depending on the duration of storage. Moldy
corn is not suitable for long-term storage.
6. Cool grain after drying, and maintain it at
3540F for the duration of the winter.
Aeration can be used for temperature control.
If the corn will be stored during the summer,
use aeration to warm it to 5060F in the
spring. Use aeration to control moisture
and temperatures during cool periods in the
summer. Next to moisture content, temperature
is the most important factor in preventing the
development of molds and toxins.
7. Control storage insects.
8. Check grain every 2 weeks in storage (more
often if quality is suspect) for temperature,
crusting, hot spots, moisture, and mold. If
any of these conditions are detected, steps
should be taken immediately to reduce the
temperature, aerate the bin, break up hot spots,
or remove spoiled grain.

If extensive infection is observed, a sample


should be collected, as described on page 1,
for aflatoxin analysis. Decisions on handling
moldy grain should be made before the field is
combined. Depending on the toxin results, the
field should be harvested as soon as possible
and the corn dried immediately to prevent
further toxin development. After harvest,
spoilage can occur quickly if delays result from
indecision.
3. Adjust the combine to minimize kernel damage.
Fungi infect damaged kernels more easily than Figure 2. Aspergillus flavus spores on damaged
corn kernels.
intact ones.
Aatoxins in Corn 3

9. Antifungal agents can be applied to grain to


reduce mold growth in storage. These products,
such as propionic acid, do not kill the mold
already present or reduce toxins already
formed. They may have disadvantages, such as
restricting use of the corn. If you plan to sell
the corn, be certain that antifungal agents
are allowable before using them.
For more details on proper grain handling to
prevent mold, mycotoxins, and other problems, see ISU
publications AED 20, Managing Dry Grain in Storage,
and Midwest Plan Service publication MWPS 13, Grain
Drying, Handling, and Storage Handbook.

What to do with aflatoxincontaminated corn


Corn that is contaminated at levels greater than
20 ppb may not be sold for interstate commerce.
However, most grain can find a safe and legitimate
use. Cleaning grain by screening or a gravity table can
reduce aflatoxin concentrations by removing the most
heavily contaminated particles. However, this can be
expensive and it is not possible to predict just how
much the aflatoxins will be reduced. The discards from
the cleaning process should not be used as feed.

Feeding contaminated grain


Aflatoxin-contaminated grain may be used
locally for animal feed, under the guidelines shown in
Table 1. Livestock producers may be willing to purchase
contaminated corn if it is below 200300 ppb. There
will probably be a discount to the price received, but
there may not be other options. It is important that a
good estimate of the aflatoxin level is obtained so that
informed decisions can be made about feeding.

Binding agents such as sodium bentonite


and aluminosilicates reduce the effects of aflatoxins on
livestock. These products are approved for use in feed,
although the FDA does not recognize their aflatoxin
management properties.
Blending aflatoxin-contaminated grain with
clean grain is not legal except in advance of direct
feeding operations. Blended grain may not be sold in
general commerce. Once aflatoxin levels are known or
suspected, it is the owners responsibility to isolate that
corn from general commerce.
Ensiling corn usually does not reduce aflatoxin
concentrations, although concentrations are unlikely to
increase in properly managed silage.

Ethanol/Wet Milling
Corn with aflatoxins can be used for ethanol
production. Aflatoxins do not accumulate in the ethanol
but will be concentrated in the distillers grains coproduct. In wet-mill processing, aflatoxins concentrate
in the gluten co-products. A rough estimate is that
aflatoxin levels in feed co-products will be four times
those in whole corn. Therefore, processors may not
accept corn with aflatoxin if their co-product markets
are sensitive to aflatoxin levels, such as dairy feed.

Ammoniation
Anhydrous ammonia reacts with aflatoxins and
reduces concentration. However, this practice is not
approved for interstate commerce, so ammoniated grain
can be used only on-farm. Ammonia can be applied as
a gas or liquid, but in either form it is a difficult and
dangerous procedure. This should be done only by a
trained and experienced operator.

File: Pest management 2-5


Prepared by Gary Munkvold, extension plant pathologist; Charles Hurburgh, professor of agricultural and biosystems engineering; and
Julie Meyer, plant pathologist.
Updated by Charles Hurburgh, professor of agricultural and biosystems engineering; Dan Loy, professor, animal science and Alison
Robertson, extension plant pathologist.

. . . and justice for all


The U.S. Department of Agriculture (USDA) prohibits discrimination in all its programs and activities on the basis of race, color, national origin, gender, religion, age,
disability, political beliefs, sexual orientation, and marital or family status. (Not all prohibited bases apply to all programs.) Many materials can be made available in
alternative formats for ADA clients. To file a complaint of discrimination, write USDA, Office of Civil Rights, Room 326-W, Whitten Building, 14th and Independence
Avenue, SW, Washington, DC 20250-9410 or call 202-720-5964.Issued in furtherance of Cooperative Extension work, Acts of May 8 and June 30, 1914, in cooperation
with the U.S. Department of Agriculture. Jack M. Payne, director, Cooperative Extension Service, Iowa State University of Science and Technology, Ames, Iowa.

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