aflatoxin risk management in texasotscweb.tamu.edu/otsc-present/2010/mycotoxin ncgb-tjh.pdf ·...

OFFICE OF THE TEXAS STATE CHEMISTTexas Feed and Fertilizer Control Service Agriculture Analytical Service

AFLATOXIN RISK MANAGEMENT IN TEXAS

&

PURSUIT OF A ONE SAMPLE STRATEGY

Tim Herrman

PhDProfessor and Director

Presented Nov. 16, 2010, for theNation Corn Producers Association

Mycotoxin

Working Group

OFFICE OF THE TEXAS STATE CHEMIST

Office of the Texas State Chemist

HistoryThe 26th Texas Legislature (1899) enacted the first Fertilizer Control Law and placed responsibility for its administration in the Division of Chemistry The 29th Legislature (1905) enacted the first Feed Control Law and located the responsibility for carrying out its statutory requirements with the Director, Texas AgriLife Research In 1984, the Texas Feed and Fertilizer Control Service (TFFCS) and the Department of Agricultural Analytical Services were combined to establish the Office of the Texas State Chemist (OTSC)


Mycotoxin regulation in TexasThe Texas Agriculture Code, Chapter 141 (Texas Commercial Feed Control Act) exempts grain (§141.002(c)(2)) or whole seed that does not contain toxins or chemical adulterants. Grain or seed exceeding the 20 ppb aflatoxinaction level is adulterated and subject to the Texas Commercial Feed Control Act. The Texas Feed and Fertilizer Control Service is the state regulatory agency responsible for administering TAC 141 and this agency is part of the Office of the Texas State Chemist (OTSC), which is in Texas Agrilife Research


MissionWhy the organization exists

Customer FocusIdentify and satisfy the customer

according to the Mission

Internal ProcessesHow to achieve outputs to

satisfy customers

Knowledge and DataInformation and skills needed

to complete internal processes

Financial ResourcesMeans of acquiring and

distributing funding to

support the other perspectives

Application of aBalanced Scorecardto align resources,data, and internal processes with the OTSC mission thatis customer focused


Mission Objective #1: Enforce compliance measures to protect consumers and enhance agribusiness. Measure #1: Percentage of completed corrective actions. Description: This measure reports the percentage of

identified licensing, product registration, labeling, consumer complaints, and sample violations that require corrective actions defined below:






satisfy customers








Science based annual work planScience based sample analysis scheduleServe as risk managersIncludes monitoring of human-animal hazards

MycotoxinsMicrobiological contaminantsHeavy metalsDioxinBSE contract and grant w/FDA

OTSC Adoption of a Science-based Approach to Risk Management


Aflatoxin Annual Plan of WorkProducts Samples per Area Total Samples

New crop corn 36 504

Bulk corn & screenings

15 210

Deer Corn 10 140

Grain Sorghums 3 42

Peanut meal 1 14

Dry dog food 5 70

Total 70 980


Biological and Other Chemical Hazards FFCS Annual Plan of Work

Products Samples per Area

Total Samples

SalmonellaRendered animal protein, oilseed meals, animal feeds, vegetable

refuse

36 504

DioxinFats, Fish meal, minerals and

mineral premixes, binders, citrus pulp

8 112

Heavy MetalsFish meal, minerals and mineral

premixes

5 56

Prohibited Mammalian Protein

29 406

Total 49 1078


OTSC Risk Management Approach






satisfy customers








OTSC Factoids2009 2010 2011

Total Budget $5,478,709 $5,488,347 $5,370,823

Feed Tonnage 17,487,064 16,815,374 -

Total Personnel 55 FTE 54 FTE 53 FTE

Feed Reserve 46% 44% 45%

Estimated market value of Texas fertilizer, animal feed and pet food ~ $10 billionOTSC receives about $500,000 in grants each year from FDA and USDAReasons for personnel reduction: increased efficiency gained from automation and new instrumentation


Risk Communication

Risk Assessment

Scientific inputs

Risk Management

Decisions involvingPolicy and values

Generic components of risk analysis

Source: FAO -Food Safety Risk Analysis


Fungal Toxins

Fungus ToxinAspergillus flavus & A. parasiticus

aflatoxin

Fusarium species tricothecenesF. Verticillioides fumonisinsF. Graminearum zearalenonePenicillium verrucosum

ochratoxins

Claviceps ergot


Mycotoxicoses of AnimalsToxin General Effect Susceptable Species

Aflatoxin Liver damage including cancer, decreased milk & egg product,

Young animals, dogs, fish, poultry, dairy cattle

Fumonisin Equine leukoencephalo- malacia, porcine pulmonary edema

Horses, rabbits, swine, breeding animals

Tricothecenes Necrosis and hemorrhage in digestive tract, depressed blood regeneration

Monogastric animals pigs, chickens and turkeys, then ruminants

Ochratoxin Kidney damage, liver damage, and intestinal necrosis and hemorrhage

Swine, turkeys and chickens decreased productivity

Mycotoxin: CAST report No 139 url

[email protected]


Aflatoxin action levelsppb Product Description

20 Corn, peanut products, cottonseed meal, and other animal feeds and feed ingredients intended for dairy animals, for animal species or uses not specified above, or when the intended use is not known

20 Corn, peanut products, and other animal feeds and feed ingredients, but excluding cottonseed meal, intended for immature animals

100 Corn and peanut products intended for breeding beef cattle, breeding swine, or mature poultry

200 Corn or peanut products intended for finishing swine of 100 pounds or greater

300 Corn and peanut products intended for finishing (i.e., feedlot) beef cattle


Fumonisin advisory levelsppm Product Description

5 Equids and rabbits (no more than 20% of diet)**

10 All other species or classes of livestock and pet animals (no more than 50% of diet)**

20 Swine and catfish (no more than 50% of diet)**

30 Breeding ruminants, breeding poultry and breeding mink* (no more than 50% of diet)

60 Ruminants >3 months old being raised for slaughter and mink being raised for pelt production (no more than 50% of diet)**

100 Poultry being raised for slaughter (no more than 50% of diet)**


DON advisory levels for animalsppm Product Description

1 finished wheat products, e.g. flour, bran, and germ, that may potentially be consumed by humans. FDA is not stating an advisory level for wheat intended for milling because normal manufacturing practices and additional technology available to millers can substantially reduce DON levels in the finished wheat product from those found in the original raw wheat. Because there is significant variability in manufacturing processes, an advisory level for raw wheat is not practical.

10 grains and grain by-products (on an 88% dry matter basis) and 30 ppm in distillers grains and brewers grains (on an 88% dry matter basis) destined for ruminating beef and feedlot cattle older than 4 months and ruminating dairy cattle older than 4 months, with the added recommendations that the total ration1 for ruminating beef and feedlot cattle older than 4 months not exceed 10 ppm DON, and the total ration for ruminating dairy cattle older than 4 months not exceed 5 ppm DON. For chickens, 10 ppm DON with the added recommendation that these ingredients not exceed 50% of the diet of chickens.

5 Grains and grain by-products destined for swine with the added recommendation that these ingredients not exceed 20 percent of their diet.

5 Grains and grain by-products destined for all other animals with the added recommendation that these ingredients not exceed 40 percent of their diet.

June 29, 2010 Nonbinding recommendationshttp://www.fda.gov/downloads/Food/GuidanceComplianceRegulatoryInformation/GuidanceDocuments/NaturalToxins/UCM217558.pdf


Preharvest Research Objective Priority Priority Stakeholder Issues/NeedsIdentification and characterization of mycotoxigenic fungi

medium Identify atoxigenic fungi strains

identify resistance high medium Need atoxigenic products Exploit fungal genomics non-toxigenic strains high Improved sampling of corn and sorghumManagement practices for mycotoxin high Management of crop to reduce mycotoxins Build a database low high Need improved host resistance develop an early warning system low high More rapid detection technology practices to reduce mycotoxins high med-low Improved remediation technologyHost plant resistance, characterization and selection of plant traits

high federal insurance program (planting late is still allowed, shouldn't be after March 15)

characterize commercial corn hybrids high Policy issues related to insurance develop corn germplasm high Insurance allows farmer to keep grain

peanut breeding to develop resistance high Lack of incentives to minimize practices to reduce aflatoxinFeed lots not regulated

Sampling and detection for mycotoxins high Loop holes in law

individual kernel mycotoxin high Lack of investment by commercial seed companies in host resistance to mycotoxins in TX

sampling designs and probability high Economics of developing resistance (2%) seed sales and 1/3 has problem

individual kernel spectral analysis high Lack of uniformity between states on surveilllanceDetoxification using binding agents and enterosorbent clays

medium high Dairy feed problem with corn shipped from midwest

mechanisms for binding Feed manufacturers having trouble getting low mycotoxin grain from midwest

Verify effectiveness high Need public corn breeders working on aflatoxin

criteria for approving binding agents

Storage management to control mycotoxins build an educational outreach program high establishing a faculty appointment lowRegulatory approaches to mitigating mycotoxin hazards in TX

high

targeted surveillance verification of sampling, testing, records monitor mycotoxin nationally

Post Harvest Research Objective


SAMPLING: Office of the Texas State Chemist Field Procedures

Samples must be obtained by a procedure which yields a representative sample using procedures of AOAC International or procedures that are determined dependable through research and/or investigation. Refer to Section 965.16 of the 15th

Edition of AOAC “Sampling of Animal Feed – Procedure.”


Risk Management Agency: Loss Adjustment Manual


% CV = sx

x 100

s = s2

( )S

1n -1

x

n2

i=1

n ii 1

h

= −

⎡

⎣

⎢⎢⎢⎢

⎤

⎦

⎥⎥⎥⎥

∑∑=xi

2

2

x=x x x ... x

nx

n

where: x = mean x x x ... individually measured value (x n = number of measurements

1 2 3 n i

1 2 3 1

+ + + +=

+ + =

∑

)

Mean

Variance

Standard Deviation

Coefficient ofVariation

1 + 2 + 33

1= 1 [14 - (6)2 ]2 3

1 = √1

50% = 1 x 1002

Sampling Statistics


Sampling Statisticss2

c(s) = aCb where a and b are constants and C is the estimated concentrationUsing regression analysis, the relationship between sampling variance and aflatoxin concentration is

s2c(s) =11.361C0.98

100 ppb aflatoxin

s2c(s)

= 11.361 x 91.2011 = 1036Standard deviation sqrt

of the variance = 32

Coefficient of variance = s ÷

μ

x 100 = 32

Johansson et al. 2000. Testing shelled corn for aflatoxin, Part I: Estimation of variance components AOACI 83(5)1264-1269


Sampling Statistics: sample sizes2

c(s) = (1.13/ns) x 11.361C0.98

where ns is the sample size in kgExample 1: 1.13 kg (2.5 lbs)100 ppb aflatoxin s2

c(s) = 1 x 11.361 x 91.2011 = 1036cv = 32

Example 2: 4.54 kg (10 lbs)s2

c(s) = (1.13/4.54) x 11.361 x 91.2011 = 256.859cv = 16

Example 3: 9.08 kg (20 lbs)cv = 11.35


Chain of custody from field to lab


Aflatoxin Analysis in Corn:

AOAC 977.16 - Sampling for Aflatoxins(Preparation for Sample Procedure)


Sampling and Sample Preparation10 pound sample collected

Entire sample ground and subdivided by Romer Mill

5 pound portion further ground by RetschMill with 1.5 mm screen

Sample passes through a number 20 sieve


Sample Grinding


Confirmation Analysis (HPLC)50 grams extracted with 70 % methanol

Aflatoxins first analyzed using Neogen ELISA test kit

Aflatoxins quantitated by PHRED (Photochemical Reactor for Enhanced Detection) fluorescence after separation on high performance liquid chromatography


OTSC Aflatoxin Analyses


Analysis Accuracy and Reproducibility Contested Samples from 2010 Corn CropOTSC TVMDL Eurofins Avg Dev RSD

648 550 660 46 0.071305 260 350 30 0.0985.5 5.3 6 0 0.048110 82 103 11 0.099574 586 750 76 0.132372 317 325 23 0.061

1340 1093 1157 96 0.07125 23 26 1 0.044

0.078

N = 24


Analysis Accuracy and Reproducibility HPLC with Aflatoxin Test Kits

A B CELISA Avg 47 329 577

RSD 0.06 0.09 0.08

Aflatest Avg 47 375 585RSD 0.07 0.04 0.08

HPLC Avg 52 378 580RSD 0.06 0.05 0.06

N = 90


Analysis Accuracy and Reproducibility Gulf Country vs OTSC

GC OTSC OTSC-GC RSD

Greenville 23 417 487 70 0.144Sherman 13 481 530 49 0.093

Greenville 487 166 0.34Sherman 530 140 0.26

Composite sample (ppb)

Individual Sample (ppb)

Facility Sample No

N = 72


Mean STD RSDGreenville 515 139 0.33Sherman 449 308 0.34

Probing sample (ppb)

Comparison between one probe and 10 probesperformed by OTSC field staff at two locations

N = 144


Post Harvest BMPs

Collect a 10 pound sample from incoming grainGrind the entire grain sample using a

Romer or Viking mill before sample reductionReduce sample using a riffler to retain

representative property of the sampleFine grind proportion for analysis to pass

through a 20 mesh sieveUse testing methods approved by either

USDA or AOACI


Impact of Particle Size on Particle Number

Percent through a 20 mesh sieve

Average Particle Size

Standard Deviation

Average Particle Number per gram

55 771 1.65 9,903

65 702 1.63 12,442

75 590 1.67 23,303

85 500 1.71 42,929

50 gram sample with 45% on top of the 20 mesh sieve = 495,150

total particles50 gram sample with 15% on top of the 20 mesh sieve = 2,146,450 total particles


Post Harvest BMPs

Store grain over the action level separately Test incoming unit trainsTest incoming grain from farm storageDo not commingle grain exceeding the

action level with grain below the action level during reclaim Correctly label grain exceeding the action

levelhttp://mycotoxinbmps.tamu.edu/


Sample Reduction for Mycotoxin Analysis


Approved Test KitsCompany Test Kit Range

ppbIdentified Commodities

NeogenCorp

Veratox-AST 5 – 300 Corn, Corn Meal, Corn Germ Meal, Corn Gluten Meal, Corn Meal, Corn/Soy Blend, Milled Rice, Popcorn, Sorghum, Soybeans, Wheat

R- Biopharm

Ridascreen FAST

5 – 300 Corn, Milled Rice, Popcorn, Corn Meal, Cracked Corn, Wheat, Soybean, Corn Germ Meal, Corn Gluten Meal, Corn Soy Blend, Sorghum

Romer FluoroQuant 5 – 300 …

Strategic Diagnostics , Inc.

Myco√ 5 – 300 …

VICAM Aflatest 5 – 300 …

http://mycotoxinbmps.tamu.edu


OTSC Test Kit ValidationGIPSA performance verification on naturally contaminated samples

Three operators will conduct all analyses. All operators shall receive up to two days of training in the use of each test kitIf an observation is a suspected outlier, the available documentation will be reviewed to identify the source of the error and make corrections where possible.


Mycotoxin MitigationAnhydrous ammonia

Procedures in the Texas Administrative Code Chapter 61, Commercial Feed Rules §.61.65

Sequestering agentsThe Office and OTSC Advisory Committee evaluating possible rule change to approve clay bindersPermit, Recordkeeping, Label, Customer formula feed, no change in action levels

OzonationOzone is a powerful oxidant capable of reaction with numerous chemical groups. After 72 hours of ozonation AFB1 level decreased 92.7%. J. M. KING, A. Moustafa, and J. N. Losso. LSU


One Sample StrategyAt the request of stakeholders during the January 11,

2010 meeting in College Station, TX, the director of OTSC began to explore the concept of using a single sample for managing aflatoxin risk at the first commercial collection point.

Firms that choose to participate in the voluntary “One Sample” program will use validated aflatoxin test kits and follow a prescribed sampling pattern, sample size, and amount of material ground tested for aflatoxin, demonstrate performance to analyze aflatoxin using OTSC check samples, use control samples, and follow record keeping procedures. It is the intention of the Service to accept commercial grain elevator results for those firms choosing to participate in voluntary “One Sample” program.