Considera*ons for Limi*ng and Preven*ng Microbial Contamina*on in Whole Grain Cereal Based Products

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Considera*ons for Limi*ng and Preven*ng Microbial Contamina*on in Whole Grain Cereal Based Products Dr. Andreia Bianchini The Food Processing Center Food Science and Technology Department University of Nebraska - Lincoln

Why Worry About Safety? ü Foodborne diseases/outbreaks v Worldwide over 1 billion cases occur per year v Over 5 million children die each year from foodborne diseases v In the US 9.4 million cases are eskmated to occur annually ü Economic losses associated with contaminated product: v ProducKon Losses v Product Recall v Product Liability v Loss of Business ü It is the right thing to do!

Wheat Flour: A Food Safety Risk Factor? Historically: Flour and grain based products have always been considered a safe product, regarding pathogens! More recently, however ü Foodborne outbreaks associated with flour: v 2005 - United States (Salmonella) v 2008 - South Island of New Zealand (Salmonella) v 2009 - United States (Escherichia coli O157:H7)

Mycotoxins of Greatest Concern in Grains Mycotoxins Molds Aflatoxins Aspergillus flavus, A. parasi*cus, A. nomius Ochratoxin Aspergillus ochraceus, A. niger Penicillium verrucosum Fumonisins Fusarium ver*cillioides (moniliforme) F. proliferatum, F. subglu*nans Deoxynivalenol Fusarium graminearum, F. culmorum (DON, Vomitoxin) F. pseudograminearum Zearalenone Fusarium graminearum, F. culmorum F. crookwellense

Challenges for the Grain Industry Microbiological quality of wheat grains Microbiological quality of milling- end products Trends in grain based foods Trends in grain processing Consumer eakng habits

Microbiological Quality and Safety of Grains Raw agricultural commodikes: ü Subject to contaminakon from the environment ü Broad diversity of microbial contaminants Microbial Test Standard* Flour** Aerobic plate count <10 5 1.1 10 5 Enterobacteriaceae/ Coliforms <100 150 E. coli (total) <100 12.8% posikve Salmonella Not detected in 25g 1.3% posikve *Ready- to- eat standards for pasta, pizza, meals, tarts, pies, cakes & pastries without dairy cream at the point of sale in UK **Data from 4,000 wheat flour samples from the US in 1993

Microbiological Quality and Safety of Grains FluctuaKon in microbial populakons ü ProducKon prackces, meteorological condinons and method of harveskng Panhandle South Central Southeast Weather Related Data 2011-2012 growing season precipitakon for Nebraska Source: USDA NASS Nebraska Field Office Districts Panhandle South Central Southeast PrecipitaKon* 8.96 15.90 18.96 Temperature* 70.2 F 70.5 F 65.7 F *Average precipitakon and temperature during the growing season

Microbiological Quality and Safety of Grains FluctuaKon in microbial populakons ü ProducKon prackces, meteorological condinons and method of harveskng Microbiological Profile* 6.0 5.0 Log CFU / g 4.0 3.0 2.0 TPC Yeast Mold Enterobactereaceae 1.0 0.0 Panhandle South Central Southeast

Microbiological Quality and Safety of Grains The majority of the microorganisms are concentrated towards the surface of the kernel ü Some species can occupy the inner part of the kernel Laca et al., 2006 DistribuKon of microbial contaminakon within cereal grains.

Microbiological Quality and Safety of Grains Bianchini et al., 2015. DON Occurrence in Grains: A North American PerspecKve.

Microbiological Quality and Safety of Grains Bianchini et al., 2015. DON Occurrence in Grains: A North American PerspecKve.

Control Measures ü Trend/Surveillance data ü Analysis of incoming grain or Supplier Approval Program ü Awareness of the current weather issues locally and globally Informed decision about where to source grain in a global market.

Cleaning

Cleaning APCs (log CFU/g) in durum wheat samples (2001 crop season): District Dirty a Clean b Montana 7.2 6.1 NW North Dakota 8.4 8.6 NC North Dakota 8.5 8.2 NE North Dakota 8.4 8.3 SW North Dakota 8.2 5.3 SE North Dakota 8.4 6.9 Region c 8.2 ± 1.3 7.2 ± 0.5 a Sample as collected. b Seived grain. c Average of 6 district values ± standard deviakon. Manthey et al., 2004.

Cleaning Effect on mycotoxins: ü Remove moldy kernels, broken kernels, and fine materials ü Remove scab infested wheat and barley: DON 5.5 19% ü In corn: Fumonisin by 26-69% ü Moldy kernel, seed or nut: Aflatoxin (40-80%)

Cleaning Bianchini et al., 2015. DON Occurrence in Grains: A North American PerspecKve.

Washing Treatment Microbial load in durum wheat aver different washing treatments: Bacteria (log CFU/g) Fungi (log CFU/g) Dry Grain (Control) 4.3a 4.9a DisKlled Water Wash 4.2a 4.9a Ozonated Water Wash (16.5 mg ozone/l) Chlorinated Water Wash (700 mg/l) 3.8b 4.7a 2.4c 4.5a AceKc acid (1%) Wash 0.5d 1.9b AceKc Acid (1%) and Ozonated Water (20.5 mg/l) Wash Dhillon, et al., 2009. 0.2e 1.7b

Tempering

Tempering Effect of ozone during wheat tempering: Flour obtained from sox wheat (Adiyaman Beyaz) Bacteria (log CFU/g) Fungi (log CFU/g) Ibanoglu, 2001. Flour obtained from hard wheat (ArjanNn) Bacteria (log CFU/g) Fungi (log CFU/g) No Treatment 4.6 3.5 5.5 4.1 Water 4.0 3.5 5.1 4.0 Ozonated Water 1.5 mg ozone/l Ozonated Water 11.5 mg ozone/l 3.4 3.0 4.7 3.7 2.5 2.4 4.0 3.1

Tempering Effect of organic acids and salt solukons during hard wheat tempering: 5.0 Aerobic Plate Counts Log10 (CFU/g) reduction 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0% 1.0% 2.5% 5.0% 2.0% 2.5% + 1% 2.5% + 2% 5% + 1% 5% + 2% 0.0 Acetic Acid Citric Acid Lactic Acid Propionic Acid Salt LA 2.5% + Salt LA 5% + Salt AA 2.5% + Salt Organic Acids

Tempering Effect of organic acids and salt solukons during hard wheat tempering: 5.0 Enterobacteriaceae Log10 (CFU/g) reduction 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0% 1.0% 2.5% 5.0% 2.0% 2.5% + 1% 2.5% + 2% 5% + 1% 5% + 2% 0.0 Acetic Acid Citric Acid Lactic Acid Propionic Acid Salt LA 2.5% + Salt LA 5% + Salt AA 2.5% + Salt Organic Acids

Milling

Dry Milling Microbial distribukon during milling of hard wheat: 3.0 Log10 (CFU/g) reduction 2.5 2.0 1.5 1.0 0.5 0.0-0.5 APC Yeast Mold Eb - 1.0-1.5 Bran Shorts First Break Fine Reduction Milling Fraction

Dry Milling FB 1 content in corn and milled frackons. Each bar represents a sample of corn and various frackons from consecukve weeks (Kawa et al., 1997)

Dry Milling Wheat, barley, rice and other cereals: DON Zearalenone Bran frackon Aflatoxin Germ Ochratoxin A

Wet Milling DistribuKon of Fumonisins in Different FracKons of Wet Milled Yellow Corn Fraction FB 1 (ppm) FB 2 (ppm) Fiber 4.2 2.6 Germ 2.2 1.1 Gluten 5.4 4.8 Starch <0.1 <0.1 Steep Water 1.2 2.5 Process Water <0.1 <0.1 Modified from Bennew et al. 1996

Wet Milling Aflatoxins Steep water Zearalenone Gluten fiber Fumonisins Germ Starch: RelaKvely free of mycotoxins

AXer Milling

AXer Milling ReducKon of microbial load in wheat flour by heakng: UpreK et al., 2010 Patent ApplicaKon Heat- treated flour. US 2010/0092639 A1..

AXer Milling ReducKon of Salmonella in corn meal by heakng: VanCauwenberge et al., 1981.

AXer Milling Effect of irradiakon from a 60 Co source on the microbial load of meals: Contaminant Wheat Meal Corn Meal Oat Meal IrradiaKon levels (Kgy) 0 1 10 25 0 1 10 25 0 1 10 25 Escherichia - - - - - - - - - - - - +++ +++ - - - - - - ++- +++ - - - - - - Bacillus +++ +++ - - - - - - +++ +++ - - - - - - +++ +++ - - - - - - Serra*a +++ ++- - - - - - - +++ +- - - - - - - - +++ ++- - - - - - - Enterococcus +++ +++ - - - - - - ++- +- - ++- - - - - - - - - - - - - - - - Clostridium +++ +++ - - - - - - +++ +++ +++ - - - +++ +++ - - - - - - Pseudomonas - - - - - - - - - - - - - - - - - - - - - - - - ++- - - - - - - - - - Micrococcus - - - ++- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - Molds +++ +++ - - - - - - +++ +++ - - - - - - +++ +++ - - - - - - Hanis et al., 1988.

Thermal Processing

Thermal Processing Effect of moisture and temperature on the inackvakon by extrusion of Enterococcus faecium as a surrogate for Salmonella: Minimum survival: 81.1 o C and 28.1% moisture Bianchini et al., 2012. ValidaKon of extrusion as a killing step for Enterococcus faecium in a balanced carbohydrate- protein meal by using a response surface design.

Thermal Processing Corn Flakes Processing Corn Grits Cooking (Steam Pressure Cookers) Hot- Air Drying Flaking ReducKons of 85% in fumonisins may be achieved. RoasKng

Thermal Processing Corn flakes processing: ü Aflatoxin: v Cooking and flaking with and without sugar 64-67% Aflatoxin v Aver toaskng with and without sugar 78-85% Aflatoxin ü

Thermal Processing 175 o C: 16% ReducKon ü Corn Muffin (5 mg/kg FB 1 ) 200 o C: 28% ReducKon (Jackson et al., 1997) Higher Loss at the Surface ü Corn Muffin (5 mg/kg FB 1 ) 204 o C: No ReducKon 232 o C: 48% ReducKon (Castelo et al., 1998)

Thermal Processing Fried TorKlla Chips Corn NixtamalizaKon (Cooking/Steeping) Masa (Rinsed and Ground) SheeKng/Cu{ng/Baking ReducKons vary from 37-78% (Voss et al., 2001) Frying

Thermal Processing ü Aflatoxin v TorKlla: 52% v TorKlla chips: 84% v Corn chips: 79% ü Zearalenone v TorKlla: 59-100% ü Deoxynivalenol v TorKlla: 72-82% Fried TorKlla Chips:

Thermal Processing ü Aflatoxin Extrusion process: v 140-185 C 33-38% ReducKon v >185 C 66% ReducKon ü Zearalenone (120-160 C) 70% ReducKon ü Deoxynivalenol (120 C) 11-27% ReducKon ü Fumonisin (160-200 ) 46-76% ReducKon ü Fumonisin (160 C, Glucose) 93% ReducKon Temperature and screw type are important!

Consumer Behavior ü In the 2009 raw cookie dough outbreak in the US: v 94% of case pakents consumed raw cookie dough v 11% of control consumers reported eakng raw cookie dough v Several pakents bought the dough with the intenkon of eakng it unbaked ü A nakonal survey in the US in 2011 revealed that consumers eat raw products occasionally: v 58% - Cookie dough v 24% - Biscuit v 22% - Pie crust v 11% - Pizza crust

Summary ü Microorganisms and mycotoxins are naturally associated with grains v Keeping them out from the food chain is the best approach ü Several factors influence the level of contaminakon and/ or mycotoxin in the final grain based product v Ingredient storage v Processing choices and parameters o FuncKonally of the flour? o Mycotoxins: True reduckon or chemical conversion? ü Consumer educakon could reduce pathogen risks associated with certain products