preliminary report, independent testing by windsor laboratories

7/25/2019 Preliminary Report, Independent Testing by Windsor Laboratories

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Windsor Laboratories894 Hawthorn Avenue Mechanicsburg, PA 17055

Phone: (717) 796-0537 Fax: (717) 796-0537

Microscopic Evaluation of Agricultural Products Members of the American Oil Chemists Society (AOCS)

July23,2014

EXPERTREPORTOFJAMESV.MAKOWSKI

I. EXECUTIVESUMMARY

ThisReportsummarizesthemicroscopicanalysesIperformedonthefollowingpetfoodstodetermine

theiringredientcontent:

BlueBuffaloLifeProtection AdultChickenandBrownRice(Dog)

BlueBuffaloFreedom AdultGrainFreeChicken(Dog)

BlueBuffaloWilderness AdultChickenRecipe(Dog)

Blue

Buffalo

Basics

Adult

Turkey

&

Potato

(Dog)

BlueBuffaloLongevity LongevityforAdultDogs(Dog)

PurinaProPlanSELECTAdultGrainFreeFormula(Dog)

PurinaONEbeyOndWhiteMeatChicken&WholeBarleyRecipe(Dog)

BlueBuffaloLifeProtection IndoorHealthChicken&BrownRiceRecipe(Cat)

BlueBuffaloFreedom GrainFreeChickenforIndoorCats(Cat)

BlueBuffaloWilderness AdultChickenRecipe(Cat)

BlueBuffaloBasics AdultTurkey&Potato(Cat)

BlueBuffaloLongevity LongevityforAdultCats(Cat)

PurinaONEbeyOndWhiteMeatChicken&WholeOatMealRecipe(Cat)

PurinaONEbeyOndSalmon&WholeBrownRiceRecipe(Cat)

Based on the microscopic analyses of these products, I conclude that several of the Blue Buffalo

productstestedcontainmeasurableamountsofchicken/poultrybyproductmealorgrains.

II. EXPERIENCEANDQUALIFICATIONS

I,JamesV.Makowski,ampresentlyemployedbyMessiahCollege inMechanicsburg,Pennsylvania,as

ProfessorofBiology. I teachcourses including,amongothers,undergraduatecoursesonCellularand

Molecular Biology, Microbiology, and Genetics (all of which include units on microscopic analysis

methods andprocedures) and conduct corresponding research. I also teach a seniorundergraduate

courseinBioethics. IearnedaPh.D.inCurriculum&Instruction/ScienceEducationattheUniversityof

Delaware, aMasterofArts inBiology atWestChester StateUniversity and aBachelorof Science in

Biologyat

Messiah

College.

Iam

the

Owner

and

Microscopist

of

Windsor

Laboratories

of

Mechanicsburg,Pennsylvania,apositionIhaveheldfor28years. Inthiscapacity,Iconductmicroscopic

analysis for clients in the agricultural industryand related industries, including thepet food industry.

AttachedtothisreportasExhibit1ismycurriculumvitaestatingmyqualifications,includingalistofall

publicationsauthoredintheprevious10years.

Iamanauthorand the leadeditorofMICROSCOPICANALYSISOFAGRICULTURALPRODUCTS (4thed.

2010),withNeilVary,MarjorieMcCutcheonandPascalVeys,publishedbytheAmericanOilChemists

PUR_000207

Case: 4:14-cv-00859-RWS Doc. #: 64-1 Filed: 09/18/14 Page: 45 of 150 PageID #: 1139


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Society. Thisbookistheleadingmanualforthemicroscopicanalysisofagriculturalproducts,including

animalproteinproducts,intheUnitedStatesandworldwide. Itisusedasaprimarytrainingresourceon

the subject of microscopic analysis of agricultural products by the United States Food and Drug

Administration (FDA). Ispeakwidelyon theuseofmicroscopy forpurposessuchas thedetectionof

prohibited protein in ruminant feed, including at workshops conducted by the FDA and National

InstitutesofHealth (NIH). Ihavealsotaughtnumerous35dayshortcoursesonusingmicroscopyto

identify components in various agricultural products for the Association of American Feed Control

Officials and the American Association of Cereal Chemists. I have instructed microscopists for the

AmericanSoybean

Association,

including

in

Jamaica

and

Trinidad.

Iam

also

the

Chairperson

of

the

TrainingCommitteefortheAgriculturalMicroscopyDivisionoftheAmericanOilChemistsSociety.

III. SCOPEOFENGAGEMENT

IwasretainedinFebruary2014byNestlPurinaPetCaretoconductmicroscopicanalysisofseveraldog

andcatfoods. Atthetimeofmyretention,IwasaskedtoanalyzeandlistallingredientsthatIwasable

to identify inthedogandcatfoodsprovidedtome. Iwasnotawareofthereasonfortherequested

work until after I completed my analysis. I was not provided with any reasons, hypotheses,

expectations, suspicions, or goals associated with my analysis. After completing my independent

analysis, Iwas informed that thepurposeof the requestedworkwas toprocurean independentand

unbiased analysis and determination ofwhether Blue Buffalo brand dog and cat foods contain any

ingredientsthat

Blue

Buffalos

marketing

materials

claim

that

they

do

not

contain,

including

chicken

and

poultrybyproductmealand,forsomeproducts,grains.

IV. SOURCINGOFANALYZEDPRODUCTS

I was provided and received from Purina by personal delivery on February 27, 2014, the following

commercially available products: (i) 10 sealed, new, unopened bags of the Blue Buffalo products

identifiedabove;and (ii)4 sealed,new,unopenedbagsofPurinadogandcatpet foodsas identified

above from Christopher Purschke at Purina. It ismy understanding that these bagswere obtained

commercially from a PetCo retail store inornearMechanicsburg. OnMarch 5,2014, I receivedby

FederalExpressanadditional14unopenedbags,duplicatesoftheoriginalfoods,fromMarkRogersat

Purina.The

purpose

of

sourcing

duplicate

products

was

to

obtain

samples

from

different

geographic

regions and production batches. I transported the bags to themicrobiology laboratory atMessiah

College,where theywere securely stored pending examination. I personally inspected each bag to

ensurethattheywerenew,unopened,andingoodcondition.

TheunopenedbagswereidentifiedandtaggedonMarch12,2014. Eachcommercialproductpackage

was randomlyanduniquelylabeledusingasimplenumberingmethod(0012014,0022014,0032014,

etc.), and matching labels were applied to product sample bags (Ziploc brand freezer bags). An

approximately 200gram sample of pet food was removed and weighed from each identified

commercialproductpackageandplacedinthecorrespondinglabeledZiplocbag,whichwasthensealed.

Afterallofthepetfoodproductpackagingwaslabeled,andthecorrespondingsampleswereplacedand

sealed

in

the

properly

identified

and

labeled

Ziploc

bags,

the

commercial

product

packaging

containing

theremainderofthepet foodsshippedtomewasreturnedtoa lockedstorage room intheMessiah

Collegemicrobiologylaboratory,whereitremainsasofthedateofthisreport. ItransportedtheZiploc

samplebagstoWindsorLaboratoriesforsecureretentionandanalysis.

V. RESEARCHPROTOCOL(PROCEDURE)

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3

ThissectionprovidesadetaileddescriptionoftheproceduresIfollowedincarryingoutthemicroscopic

analysesoftheproductsamplesidentifiedabove. Agriculturalmicroscopyisanestablished,reliabletool

fordetermining ingredientspresent in agivenproduct,particularlywhere theoriginsof theproduct

and/or its ingredientsareunknown. Byseparating theconstituentsofagivensamplebydensityand

characterizing the particles by shape, color, size, softness/hardness, texture, luster, odor and other

histological andmorphological features using amicroscope, amicroscopist can reliably identify and

quantifythoseconstituents inthesamplebycomparingthemtoknownstandards. Indeed,as Inoted

above,thistechniqueisroutinelyusedbytheFDAandothersinthefoodindustrytoidentifyingredients

infood

products.

The

microscopic

analysis

methodologies

used

to

obtain

the

results

summarized

in

this

Report are the same as those I have used for the past 28 yearswhen performing similar analyses

relatingtononlitigationprojects.

A. PreparationoftheSamplesforAnalysis

IloggedineachsamplebagbycodenumberinmylabnotebookatWindsorLaboratories,assigningeach

toanexaminationboard. Ipreparedandinspectedeachproductsampleindividuallyinordertoensure

thattherewasnoconfusionormistakeregardingthespecificsamplesbeingtested. WhenIcompleted

thepreparation,analysis,andrecordingofresultsforeachuniquelyidentifiedsample,Ireturnedunused

materialstothecorrespondinguniquelyidentifiedsamplebagandthoroughlycleanedtheareaandall

toolsemployed

before

proceeding

to

the

next

sample

bag.

In themajority of cases, the sampleswere comprised of kibble and dark bits. In all such cases, I

personally and manually separated the kibble and dark bit components, and then weighed each

componentusinganOhausE400Danalyticalbalance. Ithencalculatedandrecordedthepercentages

ofkibbleanddarkbitsinmylabnotebook.

Afterweighingeachcomponent (kibbleanddarkbits), Isubdividedthem intocombinedquartersand

oppositequartersforsamplepreparationandanalysis. Thiswasdonebyspreadingeachsampleonto

parchmentpaper,dividing intoquadrants(upper left,upperright,lower leftand lowerright)andthen

samplingthediagonallyoppositequarters. Quarteringinthiswayisanacceptedprocedureforensuring

astatistically

valid

sample.

See

MICROSCOPIC

ANALYSIS

OF

AGRICULTURAL

PRODUCTS

(2010)

at

3.

This

aliquotwas then ground in a grain flourmill to crush the particleswithout causing any damage to

cellular components, and 2 grams of the ground sample were transferred into a Coors porcelain

evaporatingdish. Allequipmentwascleanedthoroughlybeforeeachuse. Theremainderof200gram

sampleswerebagged,sealedand storedatWindsorLaboratories.

B. SampleSeparationbyFlotation:

After transferring representative2gram samples intocleanevaporatingdishes, Iadded6mlofa1:1

carbontetrachloride:chloroformmixturetotheevaporatingdishandstirredthesample thoroughlyto

allowmineral,boneandotherheavycomponentstosettle. Itransferredthematerialthatfloatedtothe

top

by

pouring

the

CCL4:CHCL3,

into

another

evaporating

dish.

The

mineral/bone

fraction,

typically

referred to as the heavy fraction,was allowed to evaporate todryness,which took approximately

thirtyminutes.

Ithenadded2mlofpetroleumethertotheCCL4:CHCL3mixturecontainingthefloatingmaterial. This

altersthedensityoftheliquid,whichcausesalessdensefractionofthesampletofloattothesurface.

Thefloatingmaterialisthenpouredoffontoadryingmedium,leavingbehindamoredensefraction. I

stirredthismixturethoroughly,transferredthefloatingmaterialtofiltermedium,andallowedittodry.

PUR_000209



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Iaddedadditional2mlaliquotsofCCHL3untilnomaterial floated. Thisprocedure resulted in3or4

densitydependentfractionsofingredientsinlightertoheavierorder. Allfractionswereallowedtodry

for at least thirty minutes. This is the accepted method of separating material by density for

microscopicexamination. SeeMICROSCOPICANALYSISOFAGRICULTURALPRODUCTS(2010)at57.

IthenweighedeachfractionontheOhausE400Danalyticalbalancetothenearest0.01g,recordingthe

results inmy labnotebook. Oncea fractionwasweighed, itwastransferredontoanew,clean,black

MasoniteexaminationboardproducedatWindsorLaboratories,onwhichtheparticleswerespreadto

ensureasingle

layer

of

material

for

microscopic

analysis.

C. MicroscopicExaminationoftheProductSamples

Imicroscopicallyexaminedeachfraction,fromheaviestto lightest,using aNikonModel76210stereo

microscopeonabotanicalboomat1020X,which isthestandardrangeforanalysisofparticlesofthis

type. SeeMICROSCOPICANALYSISOFAGRICULTURALPRODUCTS (2010) at23. Foreach fraction, I

listed and recorded all identified components inmy lab notebook and calculated and recorded the

approximatepercentageofeachcomponentusingstandardmethods. SeeMICROSCOPICANALYSISOF

AGRICULTURALPRODUCTS (2010)at57. I identifiedeachcomponentbycomparing it toareference

libraryofknown ingredientsandwithmyknowledgeofparticlecharacteristicsbasedonmyextensive

experiencewith

examination

of

similar

particles.

Iestimated

the

proportion

of

each

component

by

visuallyestimating thepercentageofeach typeofparticleviewed through themicroscope. Thiswas

repeatedforeachfractionandthetotalsforeachcomponentpresentwerecalculated.

In instanceswhere Iwas unable to identify a specificparticlewhendoing this typeof analysiswith

certainty, I transferred the relevantparticle toaNikonAlphaphotYScompoundmicroscope formore

detailed examination. Theseparticleswereplaced on a new, cleanmicroscope slide and a drop of

mountingmedium added. Themountingmediumused is a1:1:1 (v/w/v)mixtureofdistilledwater,

chloralhydrateandglycerol,whichhelps todisplace trappedair and allow some ingredientsof finer

cellular structure to be more readily observable. See MICROSCOPIC ANALYSIS OF AGRICULTURAL

PRODUCTS (2010)at5. I thenobserved theparticleunder the compoundmicroscope (100400x) to

determinecellular

structure

for

identification.

Identification

of

components

at

this

magnification

is

basedoncellularstructureratherthanoncharacteristicsoflargerparticles.

The samples from the examination boardswere retained for a period of onemonth butwere then

discardedbecausesamplesundergoanunacceptabledegreeofdecompositionbeyondthispoint.

VI. RESULTSOFTHEMICROSCOPICANALYSIS

Aspreviouslynoted,theidentitiesofthe28sampleswereblindedduringmyanalysis. OnceIcompleted

myanalysisandrecordedmyresultsinthelaboratorynotebook,thecodedlognumberscorresponding

to

each

Ziploc

bag

sample

were

matched

to

the

corresponding

numbers

placed

on

the

commercial

productpackaging. Atthatpoint,Iaddedthecommercialnamesofsomeoftheexaminedpetfoodsto

theentries in the labnotebook. Upuntil this time, Iwasunawareof thepurposeof the requested

analysisofthesamplesandofanyhypothesesbeingtested.

Ithencomparedthe ingredients identifiedbymymicroscopicanalysistothe labelsofeachdogorcat

food tested. The results ofmy testing and analysis reveal that in some instances, the Blue Buffalo

products that I tested contained poultry byproductmeal, even though the packaging indicated the

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5

contrary. ThiswasespeciallyevidentinthetwosamplesofBlueBuffaloLifeProtectionIndoorChicken

& Brown Rice Formula. In both the original and duplicate samples, I found approximately 2224%

poultrybyproductmealpresent inthekibble. Inboth instances I foundsmallfragmentsofeggshell,

rawfeatherand legscale. Further, inseveralBlueBuffalosamples, Ifoundquantitiesofgrainpresent

wherethecorresponding productswerelabeledasbeinggrainfree. Forillustrativepurposes,photos

of representative poultry byproduct and grain particles are attached in Exhibit 2, a Photographic

Appendixtothisreport. ThetablebelowsummarizestheresultsfortheBlueBuffaloproductsamples.

Discrepanciesbetweenthepackageclaimandmyresultsareboldedandhighlightedinyellow.

BlueBuffalo

Product

Component %PoultryBy

ProductMeal(Two

Samples)

%Riceand/orCorn

(TwoSamples)

PackageClaim

WildernessAdult

ChickenRecipe

(Dog)

Kibble 0%,0% 0%,0% NoPoultryByProductMeal/No

GrainsDarkBits 9%,11% 2.2%,1.5%

WildernessAdult

ChickenRecipe(Cat)

Kibble 0%,0% 0%,1% NoPoultryBy

ProductMeal/No

GrainsDarkBits 8%,5% 2.2%,2.5%

LongevityAdult

Dogs

Kibble 0%,0% N/A NoPoultryBy

ProductMeal

Dark

Bits

2%,

1% N/A

LongevityAdult

Cats

Kibble 0%,0% N/A NoPoultryByProductMealDarkBits 3%,0% N/A

LifeProtection

IndoorHealth

Chicken&Brown

RiceRecipe

Kibble 22%,24% N/A

NoPoultryBy

ProductMealDarkBits 2%,2% N/A

LifeProtection

AdultChicken&

BrownRice(Dog)

Kibble 22%,0% N/ANoPoultryBy


FreedomAdult

Grainfree

Chicken

(Dog)

Kibble 0%,0% 0%,0% NoPoultryBy

ProductMeal/No

GrainsDarkBits 2%, 2% 3%, 1%

FreedomGrain

FreeChickenfor

IndoorCats

Kibble 0%,0% 0%,0% NoPoultryByProductMeal/No

GrainsDarkBits 2%, 1% 2%, 2%

BasicsAdult

Turkey&Potato

(Cat)



BasicsAdult

Turkey&Potato

(Dog)



Thefullresultsofmyanalysesoftheproductsareprovidedonthefollowingpages.

PUR_000211



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Windsor Laboratories894 Hawthorn Avenue Mechanicsburg, PA 17055

Phone: (717) 796-0537 Fax: (717) 796-0537

Microscopic Evaluation of Agricultural Products Members of the American Oil Chemists Society (AOCS)

April21,2014

Sample:BlueBuffalo;WildernessAdultChickenRecipe(Dog);5961000270BIUBMAR0315L121

09:20

EstimatedPercent:

Kibble 96.0

DarkBits 4.0

Sample:0012014(Kibble) EstimatedPercent:

Chicken/TurkeyMeal 36

Chicken 26

TotalFat 15

TotalStarch 12

VegetablePomace 8

TotalChloridesasNaCl 0.5

DehydratedAlfalfaMeal 0.2

pH=6.0

Sample:001

2014

(Dark

Bits)

Estimated

Percent:

Flax(Linseed) 48

TotalFat 16

Starch 8

PoultryByProductMeal 9

DehydratedAlfalfaMeal 8

PeaFiber 4

RiceHulls 2

VegetablePomace 2


Ground

Yellow

Corn

0.2

pH=5.8

PUR_000212



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2.

Sample:BlueBuffaloWildernessAdultChickenRecipe(Cat);5961000140BIUBFEB0415B310620

EstimatedPercent:

Kibble 98.2

DarkBits 1.8


Chicken/TurkeyMeal 36

Chicken 26

TotalFat 15

TotalStarch 12

VegetablePomace 8



pH=6.0

Sample:0022014(DarkBits) EstimatedPercent:

Flax(Linseed) 50

TotalFat 16

Starch 9



PeaFiber 3

RiceHulls 2

VegetablePomace 2

TotalChlorides

as

NaCl

0.6

GroundYellowCorn 0.2

pH=5.8

PUR_000213



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3.

Sample:BlueBuffaloLongevityLongevityforAdultDogs;5961000802BIUBJAN0815L13110:45

EstimatedPercent:

Kibble

92.4DarkBits 7.6


Fish 28

FishMeal 18

GroundRice 16

GroundBarley 15

TotalFat 11

Starch 10

TotalChlorides

as

NaCl

0.5

pH=6.0


Flax(Linseed) 62

TotalFat 12

Starch 8


PeaFiber 5

PoultryBy

Product

Meal

2

RiceHulls 1

VegetablePomace 0.3



CalciumCarbonate Trace(


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9

4.

Sample:BlueBuffaloLongevityLongevityforAdultCats;5961000814BIUBJUN1314L13215:02

EstimatedPercent:

Kibble 93.0

DarkBits

7.0


FishMeal 32

Fish 19

TotalFat 17

GroundRice 16

GroundOats 8

GroundBarley 6


pH=6.0


Flax(Linseed) 50

TotalFat 16


Starch 10

PeaFiber 4


VegetablePomace

2

RiceHulls 1



pH=6.0

PUR_000215



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10

5.

Sample:LifeProtectionIndoorHealthChicken&BrownRiceRecipe;5961000087FEB0915J220439

EstimatedPercent:

Kibble 91.5

DarkBits 8.5


Chicken 28


TotalFat 17

GroundBarley 12

GroundRice 8

GroundOats 5

FishMeal 3

Flax(Linseed) 3

TotalChlorides

as

NaCl

0.7

Carrot 0.2

pH=6.0


Flax(Linseed) 54

TotalFat 17

Starch 10


PeaFiber

5


VegetablePomace 1

RiceHulls 1



pH=6.0

PUR_000216



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11

6.

Sample:BlueBuffaloLifeProtectionAdultChicken&BrownRice(Dog);5961000011BIUBJAN1515

T481:15

EstimatedPercent:

Kibble 95.5

DarkBits

4.5


Chicken 20


TotalFat 15

GroundRice 15

GroundBarley 10

TotalStarch 8

VegetablePomace 6

DehydratedAlfalfa

Meal

2


Flax Trace(


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12

7.

Sample:BlueBuffaloFreedomAdultGrainFeeChicken(Dog);5961000680BIUBMAR1715VS06:20

EstimatedPercent:

Kibble 93.8

DarkBits

6.2


ChickenMeal 32

Chicken 21

TotalFat 16

TotalStarch 10

Peas 8

VegetablePomace 7


Flax(Linseed) 2

TotalChlorides

as

NaCl

0.7

pH=5.2


Flax(Linseed) 57

TotalFat 16

TotalStarch 9


PeaFiber 5

PoultryBy

Product

Meal

2

GroundYellowCorn 2

RiceHulls 1


pH=5.4

PUR_000218



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13

8.

Sample:BlueBuffaloFreedomGrainFeeChickenforIndoorCats;5961000707BIUBJAN1215P1614

33K14

EstimatedPercent:

Kibble

95.7DarkBits 4.3


Chicken 32

ChickenMeal 28

TotalFat 14

TotalStarch 14

PeaFiber 6

Flax(Linseed) 2


VegetablePomace

1


pH=5.7


Flax(Linseed) 53

TotalFat 15


PeaFiber 8

TotalStarch

8


RiceHulls 2

VegetablePomace 1


pH=5.7

PUR_000219



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9.

Sample:BlueBuffaloBasicsAdultTurkey&Potato(Cat);5961000727BIUBMAY0614P173733BO5

EstimatedPercent:

Kibble 96.1

DarkBits

3.9


Chicken 35

ChickenMeal 24

TotalStarch 16

TotalFat 12

PeaFiber 6

Flax(Linseed) 2

VegetablePomace 2


TotalChlorides

as

NaCl

0.60

pH=5.5


Flax(Linseed) 54

TotalFat 16


PeaFiber 7

TotalStarch

7


RiceHulls 1

VegetablePomace 1


pH=5.7

PUR_000220



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15

10.

Sample:BlueBuffaloBasicsAdultTurkey&Potato(Dog);5961000752BIUBMAR0515P104133MO5

EstimatedPercent:

Kibble 96.1

DarkBits

3.9


Chicken 50

ChickenMeal 22

TotalStarch 12

TotalFat 12


VegetablePomace 0.2

RiceHulls 0.2


DicalciumPhosphate

Trace

(


16/46

16

11.

Sample:PurinaONEbeyOndSalmon&WholeBrownRiceRecipe;1780012702328160010110L04

EstimatedPercent:

DarkKibble 58.7

LightKibble

41.3

Sample:0112014(DarkKibble) EstimatedPercent:

ChickenMeal 24

Fish 22

TotalFat 18

GroundRice 15

SoybeanMeal 8

FishMeal 4

GroundBarley 4

GroundOats

2

SoyProteinIsolate 0.5

BeetPulp 0.5


pH=5.8

Sample:0112014(LightKibble) EstimatedPercent:

ChickenMeal 25

Fish

23

TotalFat 16

GroundRice 16

SoybeanMeal 7

FishMeal 5

GroundBarley 3

GroundOats 2


BeetPulp Trace(


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17

12.

Sample:PurinaONEbeyOndWhiteMeatChicken&WholeOatMealRecipe;1780014418

331560012305L05

EstimatedPercent:

DarkKibble

58.7

LightKibble 41.3


Chicken 27

ChickenMeal 22

TotalFat 17

GroundRice 13

SoybeanMeal 11

GroundBarley 6

GroundOats 2

SoyProtein

Isolate

0.5

BeetPulp 0.5


pH=5.7


Chicken 26

ChickenMeal 20

TotalFat

17

GroundRice 12

SoybeanMeal 12

GroundBarley 8

GroundOats 2


BeetPulp 0.3

VegetablePomace 0.1


CalciumCarbonate Trace(


18/46

18

13.

Sample:PurinaProPlanSELECTAdultGrainFreeFormula(Dog);3810015196307110820548Ll5

Kibble(0132014) EstimatedPercent:

CanolaMeal 28

Chicken

26TotalFat 18

TotalStarch 18

FishMeal 4

PeaFiber 3

NaCl 0.6

BeetPulp 0.5

pH=5.7

Sample:PurinaONEbeyOndWhiteMeatChicken&WholeBarleyRecipe(Dog);1780014936

331810850201L04


Chicken 36

GroundRice 22

TotalFat 18

CanolaMeal 12

GroundBarley 10

NaCl 0.58

CalciumCarbonate 0.1

VegetablePomace Trace(


19/46

19

14.

Sample:BlueBuffaloFreedomGrainFeeChickenforIndoorCats;BIUBSEP0514J111635

EstimatedPercent:

Kibble 94.9

DarkBits

5.1


Chicken 33

ChickenMeal 28

TotalFat 13

TotalStarch 15

PeaFiber 5

Flax(Linseed) 2


VegetablePomace 0.5

TotalChlorides

as

NaCl

0.6

pH=5.8


Flax(Linseed) 52

TotalFat 16

TotalStarch 10


PeaFiber 7

RiceHulls

2

VegetablePomace 2



pH=5.7

PUR_000225



20/46

20

15.

Sample:BlueBuffaloLongevityLongevityforAdultCats;BIUBNOV2914L13301:12

EstimatedPercent:

Kibble 90.0

DarkBits

10.0


FishMeal 20

Fish 20

GroundRice 18

TotalFat 16

GroundOats 12

GroundBarley 12


pH=6.1


Flax(Linseed) 54

TotalFat 15


Starch 8

PeaFiber 4

VegetablePomace 4

RiceHulls

2

GroundYellowCorn 1

WheatMiddlings 1


pH=6.3

PUR_000226



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21

16.

Sample:BlueBuffaloLifeProtectionAdultChicken&BrownRice(Dog);BIUBJAN1415T1C16:02

EstimatedPercent:

Kibble 91.0

DarkBits 9.0


Chicken 22

ChickenMeal 20

TotalFat 14

GroundRice 13

GroundBarley 12

TotalStarch 8

VegetablePomace 8


TotalChlorides

as

NaCl

0.51

pH=5.0


Flax(Linseed) 52

TotalFat 16

TotalStarch 10


PeaFiber 6

VegetablePomace

3

GroundYellowCorn 2

RiceHulls 2


pH=5.0

PUR_000227



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22

17.

Sample:PurinaONEbeyOndSalmon&WholeBrownRiceRecipe;400710831313L05BIUBJUL2015

EstimatedPercent:

DarkKibble 54.2

Light

Kibble

45.8


ChickenMeal 26

Fish 21

TotalFat 18

GroundRice 14

SoybeanMeal 7

FishMeal 5

GroundBarley 3

GroundOats

3


BeetPulp 0.5


pH=5.7


ChickenMeal 27

Fish

22

TotalFat 16

GroundRice 15

SoybeanMeal 8

FishMeal 4

GroundBarley 3

GroundOats 3


BeetPulp 0.1


pH=5.8

PUR_000228



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23

18.

Sample:BlueBuffaloBasicsAdultTurkey&Potato(Cat);BIUBSEP08142321933

EstimatedPercent:

Kibble 97.7

DarkBits

2.3


Chicken 35

ChickenMeal 20

TotalFat 16

TotalStarch 15

PeaFiber 7

Flax(Linseed) 2


VegetablePomace 1

TotalChlorides

as

NaCl

0.50

pH=5.5


Flax(Linseed) 55

TotalFat 15


PeaFiber 6

TotalStarch

6


RiceHulls 2

VegetablePomace 0.5


pH=5.7

PUR_000229



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24

19.

Sample:BlueBuffaloWildernessAdultChickenRecipe(Cat);BIUBMAR0215B311446

EstimatedPercent:

Kibble 96.1

DarkBits

3.9


ChickenMeal 32

Chicken 31

TotalFat 18

FishMeal 9

TotalStarch 8

RiceHulls 1



BloodMeal

0.1

pH=5.7


Flax(Linseed) 58

TotalFat 14

Starch 8



PeaFiber

4

RiceHulls 1.5

GroundYellowCorn 1



pH=5.5

PUR_000230



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25

20.

Sample:PurinaONEbeyOndWhiteMeatChicken&WholeBarleyRecipe(Dog);BIUBJAN2015

319710830939L09

Kibble(0212014): EstimatedPercent:

Chicken 35

GroundRice

24

TotalFat 18

CanolaMeal 11

GroundBarley 10

NaCl 0.61


VegetablePomace Trace(


26/46

26

21.

Sample:BlueBuffaloBasicsAdultTurkey&Potato(Dog);BIUBAPR0815P200534RO8

EstimatedPercent:

Kibble 96.9

Dark

Bits

3.1


Chicken 48

ChickenMeal 22

TotalStarch 13

TotalFat 12


VegetablePomace 0.2


DicalciumPhosphate Trace(


27/46

27

22.

Sample:BlueBuffaloFreedomAdultGrainFeeChicken(Dog);BIUBFEB0415P011733L06

EstimatedPercent:

Kibble 95.8

DarkBits

4.2


ChickenMeal 32

Chicken 25

TotalFat 15

TotalStarch 8

VegetablePomace 8

Peas 6


Flax(Linseed) 2

TotalChlorides

as

NaCl

0.7

pH=5.2


Flax(Linseed) 55

TotalFat 16

TotalStarch 10


PeaFiber 3

PoultryBy

Product

Meal

2

VegetablePomace 2

GroundYellowCorn 1


pH=5.0

PUR_000233



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28

23.

Sample:BlueBuffaloLongevityLongevityforAdultDogs;BIUBNOV2614L12301:52

EstimatedPercent:

Kibble 84.0

DarkBits

16.0


Fish 25

GroundRice 20

GroundBarley 18

FishMeal 15

TotalFat 12

TotalStarch 8


Carrots

Trace(


29/46

29

24.

Sample:PurinaONEbeyOndWhiteMeatChicken&WholeOatMealRecipe;BIUBJUL201533471083

0419L05

EstimatedPercent:

DarkKibble

51.1

LightKibble 48.9


Chicken 28

ChickenMeal 18

TotalFat 17

GroundRice 15

SoybeanMeal 12

GroundBarley 7

BeetPulp

0.2

VegetablePomace 0.2




pH=5.7


Chicken

28

ChickenMeal 20

TotalFat 16

GroundRice 12

SoybeanMeal 11

GroundBarley 9

GroundOats 1


BeetPulp 0.3

VegetablePomace 0.2


Calcium

Carbonate

Trace

(


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30

25.

Sample:BlueBuffalo;WildernessAdultChickenRecipe(Dog);BIUBFEB2515L12304:25

EstimatedPercent:

Kibble 92.0

DarkBits

8.0


ChickenMeal 35

Chicken 28

TotalFat 14

TotalStarch 12

VegetablePomace 7

Flax(Linseed) 1.5

DehydratedAlfalfaMal 0.3

TotalChlorides

as

NaCl

0.54

pH=6.0


Flax(Linseed) 50

TotalFat 15



Starch 7

PeaFiber

3

VegetablePomace 2

RiceHulls 1.5


Dirt&Grit Trace(


31/46

31

26.

Sample:PurinaProPlanSELECTAdultGrainFreeFormula(Dog);BIUBSEP2014307110821052L15


CanolaMeal 26

Chicken

25TotalFat 18

TotalStarch 18

FishMeal 6

PeaFiber 4

NaCl 0.54

BeetPulp 0.5

pH=5.8

PUR_000237



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32

27.

Sample:LifeProtectionIndoorHealthChicken&BrownRiceRecipe;BIUBAPR2415B420041

EstimatedPercent:

Kibble 80.3

DarkBits

19.7


Chicken 30


TotalFat 16

GroundBarley 12

GroundRice 7

GroundOats 6

FishMeal 3

TotalChlorides

as

NaCl

0.6

Carrot 0.1

pH=6.0


Flax(Linseed) 52

TotalFat 16

Starch 12


PeaFiber

6


VegetablePomace 1

RiceHulls 1



pH=6.0

PUR_000238



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33

PUR_000239



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EXHIBIT1

CURRICULUMVITAE

PUR_000240



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1

Messiah College-Department of Natural Sciences

Grantham, PA 17027

(717) 766-2511

Curriculum Vitae

JAMES V. MAKOWSKI

Home Address: 894 Hawthorn Avenue

Mechanicsburg, PA 17055

Office Address:

Department of Natural SciencesMessiah College

Grantham, PA 17027

Education:

1976 B.A. Messiah College (Biology)

1978-79 Trenton State University, NSF Fellowship

1980 M.A. West Chester State University (Biology)

1991 Ph.D. University of Delaware (Curriculum & Instruction/ Science

Education)

Teaching Experience:

1975-76 Messiah College, Teaching Assistant

1976-81 Central Bucks School District, Senior High School Biology

1981-86 Messiah College, Instructor in Biology

1986-1992 Messiah College, Assistant Professor of Biology

1993-1999 Messiah College, Associate Professor of Biology

2000- Messiah College, Professor of Biology

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2

While at Messiah College, I have taught the following courses in the Department

of Biological Sciences:

BIO 150 Biology I (First semester Biology majors)

BIO 102 Bioscience (Non-majors Biology)

BIO 106 Life Science for Elementary Education (Biological principles forBIO 114 Heredity & Human Affairs (Ethics based genetics course for non-

BIO 281 Genetics (Introductory genetics for majors)

BIO 313 Developmental Biology (for majors)

BIO 347 Comparative Anatomy (for majors)

BIO 381 Microbiology (for biology majors)

BIO 382 Microbiology for Health Professionals (for nursing & pre-med majors)

NSC 496 Science Seminar (Capstone course for senior majors)

IDS 101 First Year Seminar: Science & the Media

IDS 101 First Year Seminar: Science in Science Fiction

IDS 101 First Year Seminar: Science Fiction & Society

IDST 300 Genetics & SocietyBIOL189 Genetics for Health Professionals

BIOL160 Introduction to Cellular & Molecular Biology

BIOL 260 Genetics & Development

BIOL 317 Bioethics

BIOL/CHEM 495 Capstone

I also am responsible for teaching BIO 407 & CHE 407 Secondary Subject

Methods: Science for students majoring in biology and chemistry who wish to

become secondary teaching certification. In addition, I supervise these students

during their professional semester of student teaching.

Additional Professional Experience:

1986-87 Hershey Foods Corporation. Consultant in Food Microbiology,

1988 Polyclinic Medical Center, Faculty Intern to Medical Microbiology

Laboratory, 1988

1989 Quaker Oats Corporation, Faculty Intern in Quality Assurance

Lab, Shiremanstown, PA, 1989. Responsible for the development

of several new procedures for quality assurance laboratory.

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3

1987-present Owner, Windsor Laboratories a private consulting laboratory for

forensic microscopy of animal feed and human food, as well

unknown particles, etc.

Awards:

1991 Robert T Stegnar award in Science Education. Recognizes the

graduate student who has made the most significant contributions

to the advancement of science education. The University of

Delaware, May, 1991.

Publications:

Makowski, J. (1979). Nucleosomes: Intracellular chromatin packaging units.Unpublished Research Article, West Chester University, 1979.

Makowski, J. (1980). Autecology of the White-Tailed Deer (Odocoileus

virginianus). Unpublished research article, West Chester University,

1980.

Makowski, J. (1983). Concepts of Biology Laboratory Manual, Messiah College,

1983.

Makowski, J. (1988). Problem-solving in genetics: A review. Unpublished

Makowski, J. (1991). High school teachers' conceptions of genetics and their

Makowski, J. (1992). The influence of curriculum content knowledge of genetics

instruction. Journal of Research in Science Teaching, Vol 31, 2, February,

1994.

Makowski, J. (1992). A Microbiologist's View of Feed Microbiology.

Proceedings of the American Association of Feed Microscopists, Vol. 39,

1.

Makowski, J. (1996). The Use of Polarized Light Microscopy. Proceeding ofthe American Association of Feed Microscopists, Vol. 43, 2

PUR_000243



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4

Makowski, J. (1998). Feed Microscopy: An Undervalued Quality Assurance

Technique. Inform(9), 11, November, 1999.

Makowski, J. (2000). Electrophoretic Separation of Mammalian Hair Proteins as

a Means of Species Identification of Meat & Bone Meal Sources inAnimal Feeds. Inform(11), April 2000.

Makowski, J. (2006). Microscopy Techniques for the Identification of Animal

Protein Products. Inform.(17), September 2006

Makowski, J., Ed., (2011). Microscopic Analysis of Agricultural Products, 4th

Ed., Champagne, IL, AOCS Press.

Presentations:

"The Effects of Varied Instructional Strategies on Student Problem-SolvingBehaviors." Paper presented at the Annual Meeting of the Eastern

"Inhibition of Salmonella sp. in Extruded Feeds." A Paper presented at the

Annual Meeting of the American Association of Feed Microscopists,

"Hands-on Science Activities for Elementary School Students." Capital Area

Intermediate Unit Regional In-Service day, October, 1991.

"Classroom-Tested Recommendations for Improving High School Genetics

Instruction." Capital Area Intermediate Unit In-Service day, October,

"The Influence of Curriculum Content Knowledge on Genetics Instruction." A

Paper presented at the Fourth Meeting of the International Consortium

"A Microbiologist's View of Feed Microscopy." A paper presented at the 42nd

Annual Meeting of the American Association of Feed Microscopists, St. Louis,

MO, June 1992.

Identification of Feed Ingredients Using Stereo- and Compound Microscopy,

July, 1995, The University of the West Indies Veterinary College, Kingston,

Jamaica

Microscopic Identification of Antibiotics, Vitamins, and Minerals,@ July, 1996,

The University of Mexico Veterinary College, Mexico City, Mexico.

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5

"Polarized Light Microscopy and Its Applications to Feed Microscopy." A paper

presented at the 46th Annual Meeting of the American Association of Feed

Microscopists, Iowa City, IA, June, 1996.

The Role of Feed Microscopy in Feed Management,@ May, 1997, The AnnualMeeting of The American Oil Chemists Society, Seattle, WA

Differential Mammalian Hair Morphology as a Means of Identifying Meat &

Bone Meal Sources in Animal Feeds,@ May, 1998, The Annual Meeting of the

American Oil Chemists Society, Chicago, IL

Identifying Animal Protein Products in Animal Feeds, May, 2001, The Annual

Meeting of the American Oil Chemists Society, Minneapolis, MN

Identification of Feed Ingredients Endemic to Caribbean Countries, June, 2002,

The University of the West Indies, Kingston Jamaica

Identification of Feed Ingredients Endemic to Caribbean Countries, June, 2002,

The University of the West Indies, Port of Spain, Trinidad.

Feed Microscopy: Basic and Advanced Techniques, June 2003, Purdue University

Department of Veterinary Science, W. Lafayette, IN

Methods for Identification of Mammalian Proteins using Stereo and Compound

Microscopy, May 2005, American Oil Chemists Annual Meeting, St. Louis, MO

Transmissable Spongiform Encephalopathies (TSEs) in North American Elk, May2006, American Oil Chemists Annual Meeting, Chicago, IL

Chronic Wasting Disease and Potential Transmission to Humans, May, 2007,

American Oil Chemists Annual Meeting, Quebec City, Quebec

Current Policies Regarding the Ruminant Feeding Ban in Response to BSE, May,

2007, American Oil Chemists Annual Meeting, Quebec City, Quebec

Chronic Wasting Disease and Potential transmission to Humans, May 2009,

American Oil Chemists Annual Meeting, Orlando.

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6

Microscopy as a Means of Identification of Banned Animal Proteins in

Agricultural Products. June 2011, U.S. Food and Drug Administration, Bethesda,

MD.

Membership in Professional Societies:

American Genetic Association

American Association of Agricultural Microscopists

American Scientific Affiliation

American Oil Chemists Society

American Society of Microbiologists

Central Pennsylvania Microbiology AssociationEastern Educational Research Association

National Association of Researchers in Science Teaching

National Association of College Science Teachers

Offices Held:

Vice President, American Association of Agricultural Microscopists, 1995-96

President, American Association of Agricultural Microscopists, 1996-98

Division Council Representative, American Oil Chemists Association, 1996-98

Chair, Training Committee, AOCS Microscopy Division, 1996-present.

Member at Large, Agricultural Microscopy Division, American Oil ChemistsSociety, 2002-present

Chair, Agricultural Microscopy Check Sample Program, American Oil Chemists

Society 2008-2012

Committees:

1982-85 Faculty Growth and Development Team

1983-84 Faculty Growth and Development Team Secretary

1983- Teacher Education Committee

1992-98 Committee on Assessment of Student Outcomes

1992-94 General Education Committee1995-96 Committee on Review of Summer Academic Offerings

1998-2000 Ranked Faculty Affairs Committee

PUR_000246



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7

2001-2004 Senator, School of Health & Natural Sciences

2005-2008 Senator, School of Health & Natural Sciences

2006-2007 Natural Sciences Laboratory Sciences Curriculum Committee

2008-2009 Term Tenure & Promotion Committee

2011-2012 Assessment of 3rd

and 4th

Year Programs in Biological Sciences

Miscellaneous:

Taught the Microbiology component of the Basic Technical Training Course,

Pennsylvania Department of Environmental Resources (DER), October, 1990.

Attended Chautauqua Short Course in Recombinant DNA Methodologies,

California State University, San Francisco, CA, June, 1991.

Taught an Advanced Microscopy Techniques Short Course for the American

Association of Feed Microscopists, Dayton, OH, June, 1994


Association of Feed Microscopists, San Antonio, TX, June, 1995

Taught a combined Basic and Advanced Short Course in Microscopy

Techniques, the University of the West Indies, July, 1995.


Association of Feed Microscopists, Iowa City, IA, June, 1996

Taught an Advanced course in chemical spot testing for antibiotics, vitamins,

and minerals, the University of Mexico, Mexico City, Mexico, June, 1996

Served on a validation-standards setting panel for new teacher licensure by

Educational Testing Services for the Pennsylvania Department of Education, in

Camp Hill, PA, April 1-2, 1997.

Taught an Advance course in microscope techniques, with particular attention to

the use of polarized light microscopy. The American Oil Chemists Association,

Seattle, WA, May, 1997.

Taught an advanced course in ingredient identification for the American

Association of Feed Microscopists, Nashville, TN, June, 1997.

PUR_000247



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8

Hair Morphology as a Means of Identification of Mammalian Species in

Ruminant Feeds. May, 1998, The Annual Meeting of the American Oil Chemists

Society, Chicago, IL

A Short Course on the Microscopy of Marine, Poultry and Meat Products. May,

1999, The Annual Meeting of the American Oil Chemists Society, Orlando, FL

Taught a Short Course on Feed Microscopy, with Emphasis on Animal Protein

Products. November, 2001, Northern Crops Institute, North Dakota State

University, Fargo, ND.

Primary Instructor for the Advanced Agricultural Microscopy Short Course, May,

2007, Quebec City, Quebec

Taught Intensive Short course in Forensic Microscopy, University of the West

Indies, Kingston, Jamaica and Port of Spain, Trinidad, 2008

Taught Intensive Course on Agricultural Microscopy with Emphasis on Animal

Protein Products. June, 2011, Cincinnati, OH.

Alizarin Red as a means of Identifying Muscle Tissue in Ruminant Feeds. May,

2014, The Annual Meeting of the American Oil Chemists Society, San Antonio,

TX.

Married, three children

PUR_000248



43/46

EXHIBIT2

PHOTOGRAPHICAPPENDIX

Allof

the

images

below

were

taken

through

aNikon

SMZ

745T

Stereo

Microscope

and

captured

with

a

ZeissAxiocamERc5scameraat20X magnification,andweretakenoftheactualBlueBuffaloproduct

samplesanalyzedintheresearchdescribedinmyreport.

Figure1. Chickenorpoultrylegscale,fromBlueBuffaloLifeProtectionIndoorHealthChicken&

BrownRiceRecipe(kibble),samplecode0052014.

PUR_000249



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Figure2. Rawchickenorpoultryfeather,fromBlueBuffaloLifeProtectionIndoorHealthChicken&

BrownRiceRecipe(kibble),samplecode0052014.

PUR_000250



45/46

Figure3. Chickenorpoultryeggshellfragment,fromBlueBuffaloLifeProtectionIndoorHealth

Chicken&BrownRiceRecipe(kibble),samplecode0052014.

PUR_000251



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Figure4. Ricehullparticle,fromBlueBuffaloFreedomAdultGrainFreeChicken(Dog)(darkbits),

samplecode0072014.


preliminary report, independent testing by windsor laboratories

Documents