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McGillUniversity

GreenlandLivestockResearchStation&MinistryofAgricultureFoodandVetServicesLaboratory,Barbados

AEBI427BarbadosInterdisciplinaryProject

August14th,2019

JournalofSmallRuminants

‘HeatandHealthyMilk:’

DeterminingtheoptimalpasteurizationmethodofGoatandBlackbellySheepMilkfor

labellingrecommendationsattheGreenlandLivestockResearchInstitute,Barbados

Authors:

TariniGupta(tarini.gupta@mail.mcgill.ca)

MyriamDriss(myriam.driss@mail.mcgill.ca)

ElainaGrimm(elaina.grimm@mail.mcgill.ca)

Mentors:

Mr.JohnVaghaun(jv_vaughan@hotmail.com)

Mrs.PamellaWhitehall(pwhitehall@agriculture.gov.bb)

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TableofContents:

1. Abstract

2. Objectives

3. Hypothesis

4. LiteratureReview

4.1CurrentHealthTrendsandtheMilkIndustryinBarbados

a. GoatandSheepMilkindevelopingcountries

b. CurrenthealthtrendsinBarbados

c. MilkindustryinBarbados

d. Thedemandformilkworldwide

4.2ThestateoffoodsafetyinBarbados

a. Microbesinmilk

b. PasteurisationtechniquesinBarbados

c. Healthimplicationsofcontaminatedmilkonsusceptiblegroups

4.3NutrientcompositionofGoatMilk

a. Lipidsingoatmilk

b. Proteinsingoatmilk

c. AllergenCingoatmilk

d. Sugarsingoatmilk

4.4AgriculturalimportanceandsignificanceofBlackbellySheepinBarbados

a. UsageofSheepMilk

b. BlackbellysheepinBarbados

4.5NutrientcompositionofSheepMilk

a. LipidsinSheepMilk

b. ProteinsinSheepMilk

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c. MicronutrientsinSheepMilk

4.6OptimalFeeds

a. Optimalfeedsforsmallruminants

b. Goatfeedingsystem

c. Sheepfeedingsystem

d. FeedingsystematGreenlandLivestockandResearchInstitute

4.7MilksafetyandpasteurisationtechniquesinBarbados

a. Milksafetystandards

b. Milkstudyofpathogens

c. Waterasapotentialsourceofcontamination

d. QualityassuranceofRawmilk:

e. QualityassuranceatGreenland:

4.8Treatmentofmilk

a. Coldstorageinfluencesonmicrobialcompositionofrawgoatmilk:

b. Heattreatmentofmilk

5.MaterialsandMethodology

6.Results

7.Discussion

8Potentialsourcesoferror

9.Conclusion

10.Acknowledgements

11.Appendix

12.References

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1.Abstract

Despitetheoverallpopularityofpasteurizedmilk,manypeople,especiallyinruralareas,

drinkunpasteurizedmilk.ThisisthecaseformanypeopleinBarbadoswhodrinkthe

unpasteurizedmilksoldattheGreenlandLivestockResearchInstitute.Theconsumptionof

unsafeunpasteurizedmilkcanbeaseverehealthhazardtomanypeople-especiallysensitive

populations.

Thepurposeofthisexperimentwastodeterminetheoptimalconditionsfortheat-home

heattreatmentofgoatandsheepmilk(filteredandunfiltered)inordertocreatelabel

recommendationsforthemilksoldatGreenland.Wehypothesizedthatthemilkboiledatour

highertemperaturewouldyieldalowercoliformcountandthatthemilkthatisunfilteredwill

haveagreatermicrobialload.Theresultsindicatedthatsalmonellawasabsentinallmilk

samples.Therewas,however,highcoliformcountsinthefilteredgoat’smilkandnoneinthe

unfilteredmilk.Thissuggeststhatthereisanissuewiththefiltrationsystemandcleaning

protocols.Thecoliformswereeradicatedatthehigherlevelsofpasteurization(73.8degrees

celsiusfor10s).

Inordertosafeguardthesafetyoftheircustomers,Greenlandneedstoprintlabelson

theirmilkbottlesindicatingproperhomepasteurizationprotocols(i.ethatyieldedbythis

experiment)andimplementnewcleaningstrategiestoreducethebacterialload.

2.ObjectivesTheprimarypurposeofthisexperimentistodeterminetheoptimaltemperatureandtime

forpasteurisinggoatandsheepmilk(i.e.theleastlabourintensivemethodforconsumersto

achieveasafecoliformcountaccordingtoEUGuidelinesforsafemilk)andtocreatelabel

recommendationsforGreenland’sunpasteurizedgoatmilkandpotentialsaleofblackbellysheep

milk.

ThisexperimentalsoobservesthefilteredandunfilteredmilkingprocessesatGreenland,

placingemphasisontheirsanitationmeasures,sterilisationofimplements,storageofequipment

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andmilk,andthesourceofrawinputssuchaswaterandcleaningchemicals.Thisinformation

willformthebasisofrecommendationsmadetoGreenlandtoreducethemicrobialloadpresent

intheirrawmilk.

3.HypothesisSincehighheatisabletodestroycertainstrainsofbacteriathataredetrimentaltohuman

healthsuchasSalmonella,Staphylococcusaureus,andothercoliforms,heatingthemilkfor10s

asopposedto15swillbringthebacteriainthemilktoasafeconsumptionlevelwhilereducing

thelikelihoodofthemilk’smacronutrients(i.e.sugars,fatsandproteins)andheatlabile

micronutrientssuchasCobalamin(VitaminB12)andThiamin(VitaminB1)undergoing

structuralchangeanddamage.

Moreover,themilkwillbeheatedattwodifferenttemperatures:45°Cand73.8°C.Though

heatingthemilkforalongertimeatahighertemperaturemayleadtoalowertotalcoliform

count,themacronutrientsandheatlabilemicronutrientswillbemorelikelytobedamaged,

reducingthemilk’snutrientdensity.Therefore,itishypothesizedthatboilingthemilkfora

shorterperiodoftimeatthehighertemperaturewillallowthemacronutrientstostayintactwhile

effectivelykillingoffthebacteria.Additionally,thisexperimentdoesn’tpredicttheretobeany

differencesinidealpasteurisationtimesandtemperaturesforgoatandsheepmilk.

Lastly,itishypothesisedthatthemicrobialloadpresentintherawgoatandsheepmilkwill

exceedthatsuggestedbyEU'sstandardsforsafemilk(seeAppendix)assafedrinkingmilk.

Sincethisexperimentaldesignlooksatthreeprimarygroups:i.Goatfiltered,ii.Goatunfiltered

andiii.Sheepunfiltered,itisnoteworthytotakeintoaccounthowthemilkingsystems(i.e.via

filterorviahand)mayaffectthemicrobialloadpresentinthemilksamples.Itishypothesized

thatthenon-mechanicalsystemofmilkingviahandwillpresentagreateropportunityforentry

ofexternalsourcesofcontaminationinthemilkduetohumanproximitycausingunfilteredgoat

andsheepmilktohavehighertotalcoliformcountsthanfilteredgoatmilk.

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4.LiteratureReview4.1CurrentHealthTrendsandtheMilkIndustryinBarbados

a. GoatandSheepMilkindevelopingcountries

AccordingtotheFAO,cow,buffalo,andgoat’smilksarethethreemostcommonly

producedtypesofmilk,withcow’smilkbeingthemostproduced,studied,andconsumed

worldwide(BonfimdaSilva,2019).Themajorityoftheworld’spopulationlivesindeveloping

countries,particularlyinAsia.Populationgrowthandeconomicgrowthanddevelopmentarethe

maindriversofincreaseddemandfordairyproducts.Milkconsumptionhasrisenby10to20

milliontonsperyer-or1.2-1.3%or80millionmorepeoplearedrinkingmilkonadailybasis

(FAO,2008).Asaresult,dairymilkproductsproductionisincreasingexponentiallyworldwide-

risingby13%from2002-2007.Worldwide,goatandsheepmilkconsumptionisat28million

tons/yearwithworldwidepopulationofgoatsat1.05billionin2017andsheepisat1.2billion

in2017(Cannas,2019).

Theriseinpopularityofgoatandsheep’smilkisduetothemanyeconomic,social,and

nutritionalbenefitsthattheyprovidetopeople’ssubsistenceandlivelihoodsstrategiesin

developingcountries(Guo,2003).Thoughmilksafetyisaglobalconcern,itisabiggerconcern

indevelopingcountriesasthelackofadequatetechnologiesandproperinfrastructurecanleadto

spoilagelossesanddecreasedyields(Yamazi,2013).Therefore,acquiringknowledgeonproper

animalproduction,hygiene,andtechnologicalaspectsofthemanufacturingprocesscanaid

livelihoodconservationandpreventmilkspoilage(BonfimdaSilva,2019).

Moreover,theproductionofmilkfromgoatsandsheepcanalsomeetthedifferent

nutritionaldemandsofdifferentpopulationgroupswhoaremoresensitivetocow’smilk,suchas

theelderly,infants,andthosesufferingfromgastro-intestinalissuesorallergies(Verruck,2018).

Theproductionofnon-cowmilksenablestheopeningofnewmarketssuchasspecialtyand

valueaddedgoodslikecheese,fermentedmilk,yogurtandsoaps(Verruck,2018).Goatand

sheepmilkarethusabletopenetratethreedifferentmarkets:homeconsumption,specialityor

nicheinterests,andmedicalneeds(Park,2010).

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b.CurrenthealthtrendsinBarbados

Barbadosiscategorisedasasmallislanddevelopingstate,andpossesssimilartrendsto

thedevelopedworldwhenitcomestohealth.AccordingtotheGlobalBurdenofDiseases,

Injuries,andRiskFactorsstudy,mortalityratesaredominatedbynoncommunicablediseases

moresothaninfectiousones(Evaluation,2010).DespiteBarbados’highprevalenceofobesity,

overallmortalityrateshavebeendeclining.Theleadingcausesofmortalityare:ischemicheart

disease,diabetes,andcerebrovasculardiseases(allofwhichcanbelinkedtoobesityissues).For

thesediseases,thehighestriskfactorsareahighbody-massindex,dietaryrisks,andhighblood

pressure(Evaluation,2010).Ineffortstoreducetheseriskfactorshasledtopeoplesearchingfor

healthyalternativestomanyfoods,includingcow’smilk.Approximately10%and20%ofthe

populationover20yearsofagesuffersfromdiabetesandhypertension,respectively,with

prevalencesmorethandoublingatolderages(S.Sharma,XCao,2009).Themoststriking

epidemicamongtheCaribbeanpopulationisthehighprevalenceofoverweight[bodymass

index(BMI)>25kgm−2]andobesity(BMI>30kgm−2).Approximatelyhalfoftheadult

Caribbeanpopulationisoverweight,and25%ofadultCaribbeanwomenareobese(S.Sharma,

XCao,2009).

c.MilkindustryinBarbados

ThemilkindustryinBarbadosiswelldevelopedincomparisontootherdeveloping

nations,anditthushasthepotentialtoallowtheislandtobeselfsufficientinitsmilkproduction

(Baynes,etal.1999).In1997,freshmilkproductioninBarbadoswasapproximately6.3million

liters;however,thiswasnotenoughtosatisfylocaldemand,andpowderedmilkhadtobe

importedthen,asitdoesnow,tomeetthedemand(Baynes,etal.1999).A1999studycollected

milksamplesfrom25dairyfarmsinBarbadostotestthepresenceofantibacterialresiduesin

BarbadianmilksinceantibioticsareusedcommonlyinBarbadostotreatmastitisinanimals

(Baynes,etal.1999).Alongwiththepresenceofantibioticresidues,theoccurrenceof

StaphylococcusaureusandStreptococcusagalactiaeandthepHofthemilkweretested.The

studyfoundS.aureusin44percentofthemilk,S.agalactiaein16percentofthemilk,andthat

thepHofmilkinBarbadosrangedfrom6.5to6.8,whichiswithintherangeofacceptablepH

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values(Baynes,etal.1999).Additionally,nomicrobesorresiduesweredetectedinmilksamples

thathadundergoneultrahigh-temperaturepasteurization(Baynes,etal.1999).Thisdatashows

howmilkservesasanenvironmentinwhichbacteriacangrow(anddoesgrowinBarbados,as

wellasotherplaces),anditalsosuggeststheefficacyofpasteurizationinensuringthesafetyof

milkconsumption.

d.Thedemandformilkworldwide

Theworld’spopulationisincreasing.Withthis,theconsumption,production,and

demandformilk-whichischeap,energydense,nutrientdense,andaccessibletoruralandurban

facetsoflife-isontherise.Withthis,isariseinlivestockproduction(Speedy,2003).This

increaseinlivestockproductionisparticularlyacuteindevelopingcountrieswherethe

productionofpoultry,pigs,eggs,andmilkseesthelargestgrowth(Speedy,2003).Barbados,too,

isseeinganincreaseinitsproductionandconsumptionofanimalproducts,includingmilk

(Speedy,2003).Theincreaseinlivestockproduction,includingtheproductionofmilk,in

Barbadosanddevelopingnationsgenerallyisbeneficialasitimproveshealthconditionsby

providingkeyproteinsandmicronutrientstothedietandmakesproductiveuseofavailablefarm

resourcesthatmightotherwisenotbeused(Speedy,2003).Thisincreaseinmilkproductionis

notonlybeneficialforthestateoffoodsecurityofBarbadosbutalsohelpsreducethecountry’s

relianceonimportedmilkpowdertomeetthedairydemand(Speedy,2003).

4.2StateoffoodsafetyinBarbados

a. Microbesinmilk Theimportanceoffoodforhumanhealthiswidelyrecognised.Eatingoptimalquantities

ofnutritiousandsafefoodisapreconditionforahealthylife.Thesafetyoffoodsuchasmilk

thatisfromanimaloriginisimportantasalargemajorityoffoodborneillnessesarederivedfrom

poultry.Farmanimalsthusplayacentralfactorinhumanhealth(Buncic,2006).

Variousmicrobialgroupscancontaminatemilk,whichcanimpactthemilk’squality,

safety,andevenbeneficialfeatures(Nero,A.2019).Whiletheemphasisofmilksafetyisonthe

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eliminationofpathogeniccoliforms,itisinterestingtonotethatmilkcontainscertainmicrobes

thatmaybebeneficialtohealth.Forexample,somelacticacidbacteria(LAB)areableto

producebacteriocins,whichareantimicrobialpeptidessynthesizedintheribosomes(Cotteret

al.,2005,Molloyetal.,2011).Theycanproduceawidevarietyofbacteriocinswhicheachdiffer

inaminoacidcomposition,biosynthesis,transport,andmodeofactionandprovideprobiotic

benefitssuchasenhancedgutmicrobiotaandinturnenhanceddigestionandnutrientabsorption

(Cotteretal.,2005,Molloyetal.,2011).

Whilethebeneficialmicrobesarenaturallyoccurringinmilk,mostpathogenic

microorganismsmayentermilkintwodifferentways:eitherfromdiseasesthataffectthedairy

herd,suchastheetiologicagentsexcretedbymilk(MycobacteriumbovisandBrucellaabortus)

orthroughunhygienicmilkingandhandlingpractices.PathogenssuchasStaphylococcusaureus,

Campylobacterspp.,Salmonella,Listeriamonocytogenes,Shigatoxin-producingE.colimaybe

introducedintotheequipment,floor,soil,water,andfecesandcontaminatemilk(M.Perin,G.

Pereira,2017).

b.PasteurisationtechniquesinBarbadosThedairysectorwasoneofthefirstfoodsectorstointroduceprocessingstepsaswellas

chemicalandmicrobialcriteriaalongtheprocessingchaininordertomonitorandsafeguardthe

qualityoftheprocesseddairyproducts.Theeliminationofpathogensthroughpasteurisationhas

beenthemainfocusoftheaforementionedsafetyoperationsinthedairyindustry.Commercial

milkprocessingplantssuchasPinehillDairyinBarbadosexecuteUltraHighTemperature

(UHT)pasteurisationi.e.coolingthroughtheentiremilkchaintopreventoutgrowthofspoilage

bacteria,ultrahightemperaturetreatment(UHT)andindustrialsterilisationtoproduce

commercialsterility.Recently,themostcommonlyusedheattreatmentsarehigh

temperature/shorttime(HTST)pasteurisation,extendedshelf-life(ESL)pasteurisationand

commercialsterility(Gonzales-Barron,Gonçalves-Tenório,2017).Bydefinition,milkthatis

pasteurizedhasreceivedaheattreatmentsufficienttominimisehealthhazardsarisingfrom

pathogenicmicroorganismsassociatedwithrawmilkaswellasadversechemical,physicalor

organolepticchanges(Gonzales-Barron,Gonçalves-Tenório,2017).

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InBarbados,PinehillDairy’sultrapasteurisedmilkdominatesthedairymarketformilk

consumers,yettheconsumptionofunpasteurisedmilkissignificant.AccordingtoGreenland

LivestockandResearchInstitute,thehighestpercentageofmilkconsumersofrawgoat’smilk

fromGreenlandaretheelderly(60yearsandabove).Moreover,theInstitutesurveyedgoatmilk

consumersregardinghowtheytreatrawmilkpriortodrinking;ifanything,itwasfoundthati.

mostconsumersdonotcarrycoolerstopreventmicrobialspoilagewhiletransportingthemilkto

theirhomes,ii.mostelderlypeoplewhobuythemilkareunawareofthehealthhazards

associatedwithconsumptionofrawmilkandiii.alargeproportionofthemiddle-agedand

youngmilkconsumersreportthattheydon’tpasteurisethemilkatallpriortoconsumptionas

theybelievethatthenutritionalstatusofthemilkwilldwindlewithheattreatment.Thelackof

awarenessaroundthehealthrisksassociatedwithrawmilkmayleadtotheingestionof

pathogeniccoliformssuchasE.coli,Salmonella,Staphylococcusaureusetc.

Inordertosafeguardagainstpossibleillnessassociatedwithrawmilkconsumptionat

home,milkmustundergoaformofheattreatment.Itisespeciallyimportanttopasteurisemilk

forpeoplesusceptibletofoodborneillnessessuchaspregnantwomen,youngchildren,the

elderly,thoseinflictedwithHIV/AIDSandotherimmunesystemdiseases(Gonzales-Barron,

Gonçalves-Tenório,2017).Yet,asGreenlandreported,thelargestproportionofrawgoatmilk

consumersaretheelderly.ThisexperimentfollowsOregonStateUniversityExtensionService’s

HomePasteurisationofRawMilkGuidelinewhichsuggestsusingthedoubleboilermethodto

heatmilkforthebestqualitypasteurisationi.e.onethatwillensurethelowestfinalcoliform

count.TheuseofindirectheatalsoensuresleastdamagetoheatlabilenutrientssuchasVitamin

B12andmilkproteinslikeα-lactalbuminandβ-lactoglobulin(Gonzales-Barron,

Gonçalves-Tenório,2017).

b.HealthimplicationsofcontaminatedmilkonsusceptiblegroupsMesophilicorganismsthriveintemperaturesbetween25°Cto40°C(L.A,Nero,2015).

Thesearealsooptimalconditionsforspoilageandpathogenicbacteria.Mesophilicbacteria

commonlyassociatedwithrawmilkareMicrococcus,Staphylococcus,Enterococcus,

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Escherichia,Serratia,Acinetobacter,Flavobacterium,Pseudomonasetc.amongothers(L.A,

Nero,2015).Itiseasierforthesepathogenstothriveinthewarmconditionsofthetropics.

Moreover,multiplerecentepidemiologicalstudiesshowthatrawmilkcaneasilybe

contaminatedbypathogens,thatmayhavenegativeshorttermandlongtermimplicationson

health(P.Oliveretal.,2009).Yet,thenumberofpeopleconsumingrawmilkisontherise(P.

Oliveretal.,2009).

E.coli0157isacommonpathogenlinkedtohealthhazards.Itistheleadingcauseof

diarrheainchildrenfromlesserdevelopedcountries(Estrada-Garcia,Hodges,2013).Itmay

causetoxico-infections,producingentericand/orsystemicillnesses.Furthermore,itcanhave

severeconsequencessuchashemorrhagiccolitis,hemolyticuraemicsyndrome(HUS)and

thromboticthrombocytopenicpurpura(Estrada-Garcia,Hodges,2013).Thegastrointestinaltract

offarmanimals,especiallycattleisacarrieroftheshigatoxinproducingE.coli(L.A,Nero,

2015).Itisestimatedthatupto10%ofpatientswithShigatoxinproducingE.coli(STEC)

infectionmaydevelopHUS,withacase-fatalityraterangingfrom3to5%(L.A,Nero,2015).

Overall,HUSisthemostcommoncauseofacuterenalfailureinyoungchildren.Itcancause

neurologicalcomplications(suchasseizure,strokeandcoma)in25%ofHemolyticuremic

syndrome(HUS)patientsandchronicrenalsequelae,usuallymild,inaround50%ofsurvivors

(WHO,2018).

Moreover,drinkingrawgoatmilkandcheesehavebeenassociatedwithsalmonellosis

outbreaksinhumans.InalargeoutbreakinFrancein1993,consumptionofcheesefrom

unpasteurizedgoatmilkledtoaSalmonellaParatyphiBoutbreak;thebacteriawasisolatedfrom

milkattheprocessingplanton2of5occasionsandwasfoundinthemilkfromonly1outof40

farmsthatsuppliedtheplant(DeBuyseretal.,2001).Salmonellatyphi,astestedinthis

experiment,ispassedtohumansmainlybyingestionofcontaminatedfood,mainlybeef,pork,

poultry,dairyproducts,eggs,andfreshproduce(Hald,2013).Salmonellapenetratestheintestinal

epithelium,possiblyproliferatinginmacrophages,passintolymphnodes,liverorspleenand

whichcausessepticaemia.Furthermore,itisoneoftheleadingcausesofgastroenteritisand

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bacteremiaworldwide.Symptomsincludefever,aprolongedperiodoffatigue,andthe

developmentofgastrointestinalillnesswithacutediarrheaasthemainsymptom(Hald,2013).

Long-termirritablebowelsyndrome(IBS)isanoutcomeofsalmonellosis,whichisresponsible

forconsiderablemorbidityandhealthcarecosts(Hald,2013).Italsocauseslongtermreactive

arthritiswhichcompromisesone’squalityoflifeduetothelimitationofone’sjoints.Theelderly

aremoresusceptibletothenegativeconsequencesoftheaforementionedillnessescausedby

Salmonella.InthecaseofBarbados,whichhasalargeelderlypopulation,rawmilk

contaminatedwithSalmonellaposesamajorthreat.

4.3NutrientcompositionofGoat’sMilkFig1.1:Tabledepictingmacronutrientcomposition,energydensityandcalciumcontentin1cup(242mL)servingofcow,goatandsheepmilk:

MilkType Energy(kcal/234mL)

Sugars(g/234mL)

Protein(g/234mL)

Fat(g/234mL) Calcium(mg/234mL)

Cow 149 12 8 7.9(4.9ofwhichsaturated)

290(29%DV)

Goat 168 11 9 10(7ofwhichsaturated)

327(33%DV)

Sheep 265 13.1 14.7 17.2(11.3ofwhichsaturated)

473(47%DV)

Whilegoatmilkandcowmilkaresimilarinnatureandstructure,theypossesssome

slightdifferenceswhichcanimpactthefinalproductsofthedifferenttypesofmilkaswellas

theirdigestibilityandnutritionalvalue(aminoacidcomposition,milkproteins’secondary

structure,andotherchemicalpropertiesofthemilk)(Vanderpas,2003).Goatmilkismore

digestible,alkaline,andhasagreaterbuffercapacitythancow’smilkwhichmakesitamore

nutritiousdietarystaple-especiallyforyoungerorolderpopulations(Vanderpas,2003).Goat’s

milkconsumptionhasbeenproventoreducetheriskofcardiovasculardisease,inhibitcancer

formation,andimproveimmunefunction(Verruck,2018).Additionally,goat’smilkcontains

“bioactivepeptides,conjugatedlinoleicacidsandoligosaccharides–whichaidsinthe

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processingofthemilk”(Verruck,2018).Thesebeneficialpropertiespresentingoat’smilkare

especiallyhelpfultothosesufferingfrommilkallergies,gastro-intestinalproblems,infants,and

theelderly(Vanderpas,2003).Thepresenceofthesehealthbenefitscanaidinthedevelopment

andmaintenanceofmetabolic,immunological,andphysiologicalprocessesinhumans(Verruck,

2018).

a. Lipidsingoatmilk

Thelipidsingoat’smilkareprimarilycomposedoftriglycerides(98%oftotallipids)

withasmallamountofphospholipids(1%)andfattyacids(1%)(Verruck,2018).Thelipidsare

relativelysmallbutnumerousfatglobulesthatdonotaggregatenaturally.Thefattyacid

compositionofgoat’smilkiscomposedofshortchains–whichaidsindigestibility(Guo,2003).

Theseshortchainsareformedthroughthepolymerizationofacetateproducedbyrumenbacteria

(Verruck,2018).Thefattyacidsarecomposedofconjugatedlinoleicacid–whichisanessential

fattyacidthatislinkedtoadecreasedriskofcardiovasculardiseaseandincreasedbrainfunction

(Verruck,2018).

b.Proteinsingoatmilk

Proteinsandaminoacidsingoat’smilkarecomposedofcaseinsandwheyprotein(Guo,

2003).Caseinproteinsaresmallerandmoreabundantingoat’smilkascomparedtocow’swhich

againaidsindigestibility(Guo,2003).Theproteinfragmentsoraminoacidsingoat’smilkalso

bindbetterandexertcertainbioactivepropertiesthataidinconservinghumanhealth–thisisnot

observedincow’smilk(Verruck,2018).Moreover,goat’smilkcontainshigheramountsof14of

the18essentialaminoacids.Oftheremainingfour,tryptophan,asparticacid,glutamicacidand

glycine,onlytryptophanisanessentialaminoacid(Verruck,2018).

c.Allergeniccapacity

Bovinemilkallergiesoccurduetopoordigestibilityofthebovineproteins(Verruck,

2018).Thus,goatmilk’shighdegreeofdigestibilitydecreasesitsallergeniccapacity.

Additionally,immunoglobulinG1andhistaminelevelsarelowerwhengoat’smilkisgivento

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milk-sensitizedmice(Verruck,2018).Moreover,alpha-s1-caseinprotein,themajorproteinand

allergenincow'smilk,isabsentingoat’smilk(Verruck,2018).

d.Sugarsingoatmilk

lactoseislessprevalentingoat’smilkthanincow’s-9gramspercupcomparedto12

gramsincow'smilk(Guo,2003).Additionally,goat’smilkseemstobeformedbyseveral

polypeptidesthatcontainseveralnutritionalbenefits,suchaspromotingthegutmicroflora

(Verruck,2018).Thebetterdigestibilityandhealthbenefitsofthegoat’soligosaccharidesare

mostlikelyduetothefactthattheyarebiologicallysimilartothosefoundinbreastmilk

(Verruck,2018)whichmakesthemmoreidentifiabletoinfantsandthehumanbodyisbetterable

touseandmakesenseofthesepolypeptides.Moreover,recentstudiesindicatethat

oligosaccharidesincaprinemilkrangedfrom250-300mg/mLwhichis4-5timeshigherthanthat

inbovinemilk(W.Park,F.W.Haenlein,2013)..Milkoligosaccharidesareespeciallybeneficial

forinfantsastheyareaneasytodigestsourceoffiberandhaveantimicrobialproperties.

Additionally,sialicacidfoundinoligosaccharideshaveshownimprovementsindevelopmentof

infantsbrains(W.Park,F.W.Haenlein,2013).

Goat’smilk,likecow’smilk,isalsoconsideredtobeafunctionalfood(afoodthat

providesadditionalhealthbenefitsbeyonditsnutritionalvalue)sincegoat’smilkcanbe

fermentedintocheeseandyogurt,twofoodsthatarerichinbioavailableprobioticsto

humans(Verruck,2018).Probioticsaidinrestoringthebalancebetweenthegoodandbad

bacteriapresentintheintestinalflora,maintainingahealthygutmicroflora(Verruck,2018).

4.4AgriculturalimportanceandsignificanceofBlackbellySheepinBarbados

a. UsageofSheepMilk

Raisingdairysheeprequiresahighdegreeofmanagementskill.Itinvolvestwo

productionsystems–oneforthesheepandoneforthemilk(Hale,2006).Marketingsheep’s

milkalsopossessuniquechallenges,oneswhichmaybeamplifiedinBarbados.IntheUnited

States,sheepmilkcheesewasnotcommonlysolduntil20yearsago,anditisstilllargely

consideredanichemarket(Hale,2006).However,themarketisevolvingandnowsheep’smilk

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isbeingtransformedintootherproductssuchasyogurt,icecream,soapandcheesessuchas

Manchegothatconsumershavecometolove(Hale,2006).InBarbados,however,thereare

culturalandknowledgebarrierstothedrinkingofsheepmilk.Accordingtoboththeemployees

atGreenlandandtheemployeesattheMinistryofAgriculture’sVeterinaryServicesLab,the

consumptionofsheep’smilkisnotsomethingonewouldseeasadesirableoption.Thislackof

culturalacceptancecansimilarlybeseeninNorthAmericancountries,despitethechanging

landscapethere.

b.BarbadosBlackbellySheep

TheBlackbellysheepprovestobeahighlyprolificandimportantbreedforthe

Barbadianlandscape.TheBlackbellysheephastraditionallybeenusedasasourceofmoneyand

meatforpeasantfarmersinBarbados,andthebreedcontinuestobeimportantthankstoits

specialcharacteristics(Rastogi,R.K.,etal.).Forone,itisspecificallyadaptedtotheproduction

systemsinthetropics.Tropicalconditionsarecharacterizedbyalongdryseasonthatcanbe

pronetodroughtsandpoorpasturequalityandthereforeseasonalweightloss(Almeida,2017).

Additionally,pasturesarecomposedessentiallyoffast-growinggrasses,highinfibreandlowon

protein,andinthecaseoftheCaribbean,thebestsoilsweretraditionallyusedforsugarcane

plantationsandanimalproductionwasdoneessentiallyonroadsidesandpoorerterrains

(Almeida,2017).Finally,animalsaresubjectedtodifferenttropicaldiseasesandparasitosisthat

hinderanimalproductionintheregion(Almeida,2017).TheBarbadosBlackbellysheepdeals

wellwiththeseconditionsastheyarehighlytoleranttoparasiticinfestation,havegood

motheringinstincts,andhave“lowfatonthelongissimusdorsi(rib-eye)muscle”(McClean,et

al.2011).Thesetraitsareimportantastheyare“characteristicsofhighprolificacy”(McClean,et

al.2011).TheBarbadosBlackbellysheepisanextremelyadaptableanimalthatisabletothrive

inthetropicalanddifficultconditionsofBarbados(Almeida,2017).

Ina2014study,thepreferencesof78farmersinMacedoniaandNorthernGreecewere

testedtodeterminewhatthegoalsofsheepbreedingprogramsare.Thegoalsofbreeding

programsareimportantastheygivefarmersthecharactericsheepthatwillmaximizetheir

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profitsandtheirutility(RagkosandAbas,2014).Theresultsshowedthatfarmershaveahigh

variabilityinwhattraitstheyarelookingforintheirsheepdependingonthefarmers’needsand

thecharacteristicsofthefarmsuchasacreageandherdsize(RagkosandAbas2014).For

example,longerlactationperiodsandhighfertilityratesaredesiredbyfarmerswithlargeherds

asopposedtothosewithsmallherds(RagkosandAbas2014).Extensivefarmers,whouse

animalswithshortlactationperiodsastheyusetoomuchlandtoworkyearroundefficiently,

wantsheepwithhighermilkproductionsothattheycangetasmuchmilkoutoftheirlimited

seasonaspossible(RagkosandAbas,2014).Sheepwithlowfatdepositsarepreferredbythe

majorityoffarmers-thoughnotallsincedesireisvariablewhichisexemplifiedbythefactthat

highlandsheepfarmersactuallyprefersheepwithfatcoverage(RagkosandAbas,2014).Higher

milkproductionrates,however,aretypicallyseenasapositivefeature(RagkosandAbas,

2014).Bythenatureofthevariabilityoffarmers’preferences,diseaseresistanceandlowfaton

themusclearenotuniversallydesired,butinthetropicalconditionsofBarbados,theseareseen

aspositivefeaturesasdiseaseincidenceismuchhigherintropicalclimates(RagkosandAbas,

2014).TheBarbadosBlackbellySheepisimportantinBarbadianagricultureasthetraitsthat

allowthemtobeproductiveintheBarbadianenvironmentaresoughtafterandcarefully

preservedbyBarbadiansubsistencefarmerswhoneedproductivesheep(McClean,etal.2011).

TheBarbadosBlackbellysheepisprimarilyraisedformeat,butthesesheepareprolific

producersofmilkandcouldbeapotentialnewsourceofmilkfortheislandoncemoreisknown

aboutsheep’smilkanditismorecommonlyconsumed.

TheBarbadosBlackbellysheepisimportanttotheagriculturallandscapeofBarbadosas

itiswellsuitedtotheenvironmentandcanthusberaisedwellandutilizedwidelyinBarbados,

butthesheepbreadisalsogainingpopularityamongothernationsbecauseofitspositive

characteristics.TheBlackbellysheep’sresistancetomajordiseasesandinternalparasites,

coupledwiththefactthatthebreedhasahighprolificacyrateandcanbirthcontinuously

throughouttheyearwithoutseasonalrestriction,makethesheepapopularbreedtoimport

(Rastogi,R.K.,etal.).TheBarbadosBlackbellysheephasbeenexportedtomanyCaribbean

islandssuchasSt.Lucia,Antigua,TrinidadandTobata,andGuyanaaswellascountriesfurther

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awayfromBarbadossuchasVenezuela,Mexico,Panama,Taiwan,Canada,andtheUnited

States(Rastogi,R.K.,etal.).Forexample,thereisnowasizablepopulationofsheepinTexas

thatarecrossesoftheBarbadosBlackbellyandtheRambouilletsheep(Rastogi,R.K.,etal.).

ExportsoftheBlackbellysheepwerestoppedfrom1974to1976bytheBarbadiangovernment

inanefforttobuildupnationalnumbers,butexportoftheanimalspickedupafterthebanwhen

theirexportvaluewasseenandtheBarbadosSheepFarmersAssociationwasformedin1975

(WilliamsandYoussef).TheexportoftheBlackbellysheepinturnincreasesitsimportanceto

Barbadosasitalsoservesasasourceofadditionalrevenue.

4.5NutrientcompositionofBlackbellySheepMilk

Sheepmilkhasahigherspecificgravity,viscosity,andrefractiveindexthancow’smilk

sinceithasagreatertotalsolidscontentinthemilk,andthiscangreatlyinfluencethetasteand

textureoffinalproducts-suchasthefirmnessofayogurt(BonfimdaSilva,2019).The

macronutrientprofileofsheep’smilkissimilartothatofgoat’smilk.However,itdoescontain

higherlevelsoflactoseandlowerlevelsoffatandprotein(BonfimdaSilva,2019).Similarlyto

goat’smilk,thelipidsarepresentintheformofsmallandnumerousglobules.Theirsmallsize

canaidindigestibilityandefficacyoflipidmetabolismwhenconsumed(BonfimdaSilva,

2019).Additionally,thefattyacidprofileiscomposedofshorterchainswhicharealsoeasierto

digest(BonfimdaSilva,2019).Nutritionallyspeaking,sheep’smilkalsohashighlevelsof

VitaminsAandBbutlowlevelsofVitaminEandfolicacids(BonfimdaSilva,2019).

a. LipidsinSheepMilk

Theaveragefatglobulesinsheepmilkarethesmallestamongstsmallruminants,makingit

moreefficientintermsofdigestionandlipidmetabolism(W.Park,F.W.Haenlein,2013).

Moreover,sheep’smilkhasahigherproportionofshortchaintriglycerides(SCTG)andalower

proportionoflongchaintriglycerides(LCTG)asopposedtocow’smilk.Thishasimplications

onthephysicochemicalpropertiesofsheepmilk-itislessenergydense,hasalowermeltingand

boilingpointthancow’smilk.Themedicinalandnutritionalvaluesofthesefattyacidsareunder

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research,thusfarhavingdemonstratedbenefitsindifferentdiseases(W.Park,F.W.Haenlein,

2013).

Additionally,oleicacid,thesecondmostpredominantfattyacidinsheep’smilk,isananti

atherogenicagentanddecreasesbloodLDLcholesterollevelwhichislinkedwithahigher

incidenceofcardiovasculardisease(CVD)(W.Park,F.W.Haenlein,2013).Sincetheleading

causeofmortalityinBarbadosisischemicheartdisease(Evaluation,2010),sheep’smilk

consumptionmaybeabeneficialdietaryadditioninordertoimprovebloodlipidprofile.

Anotherinterestingconsiderationregardinglipidcompositioninsheepmilkisit’s

conjugatedlinoleicacid(CLA)content,whichishigherthanthatingoatandcow’smilk(W.

Park,F.W.Haenlein,2013).CLAcompeteswitharachidonicacidinabiochemicalpathwayand

thusreducestheconcentrationofpro-inflammatoryagentslikethromboxaneandprostaglandins.

Besidesbeinganti-inflammatoryandthusreducingtheriskofCVD,CLAisassociatedwithlean

bodymassenhancingpropertieswhichisofimportanceinreducingtheriskofobesity(W.Park,

F.W.Haenlein,2013);noteworthyinBarbados.

b.Proteinsinsheepmilk

Enzymatichydrolysisofmilkproteinscanreleasebioactivepeptideswhichaabletoexert

specificbiologicalactivities,suchasantihypertensive,antimicrobial,opioid,antioxidant,

immunomodulatory,ormineralbinding(Mine,Shahidi,2005).Sheepderivedbioactivepeptides

areknowntoprovideanonimmunediseasedefenseandcontrolofmicrobialinfections.

Angiotensin-ConvertingEnzyme(ACE)peptidesareasignificantclassofsheepmilkproteins.

Theyarewelldocumentedforpotentialbeneficialeffectsinthetreatmentofhypertension,which

isoneoftheleadingcausesofmortalityinBarbados(refertosection2.1).Recentstudiesfound

thatovinecaseinfractionsandtheirhydrolysatesareabletoexertantioxidantactivity(W.Park,

2009).Antioxidantsfightfreeradicalsgeneratedinthebodyandthereforereducetheriskof

RNA,DNAorproteindamageandfaultymetabolismandtherebyreducetheriskofvarious

cancers.

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Additionally,thetotalproteincontentinsheepmilk-14.7g/cupissignificantlyhigherthan

thatfoundincowandgoatmilk-8g/cupand9g/cuprespectively(refertoFig1.1).Achildis

almosttwotimesmorelikelytomeettheirproteinnutritionalneeds(RDA)bydrinkingoneglass

ofsheep’smilkasopposedtogoatorcowmilk.

c.MicronutrientsinSheepMilk

Fig1.2:depictingmeanandmediandailyenergyintakeofBarbadianchildren(Gaskinetal.,2019).

Thelevelsofcalcium,Phosphorus,Magnesium,Zinc,IronandCopperarehigherin

sheepthanincowmilk(W.Park,2009).A1cupservingofsheepmilkcontains47%ofthe

Calciumrecommendeddailyallowance(RDA)whichgoatandcowmilkprovide33%and27%

oftheCalciumRDArespectively.ThisisofspecialimportanceinBarbadosasmostofthe

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populationfallsbelowtheRDAforCalcium(Fig1.2),whichmayposenegativehealth

implicationssuchasahigherriskofosteoporosisorhipfracturesduetoinadequatebone

mineralisation.

4.6Optimalfeedsforsmallruminants

a. Optimalfeedsforsmallruminants

Smallruminantssuchassheepandgoatspresentuniquefeedingchallengesduetotheir

widecapacitytoadapttoavarietyofnutritionalandenvironmentalcontexts(Cannas,2019)–

theseruminants(sheepandgoats)areraisedthroughavarietyof“farmingandfeedingsystems,

diverseecologicalbackgrounds,breeds,populationsandcrosses”(Cannas,2019)duetotheir

largecapacityforadaptation.Howeverduetothislargecapacityforadaptation,theproduction

efficacyofthesheepandgoatcanbedifficulttopredictinthelongrun(Cannas,2019).Feed

recommendationsareusuallybasedaroundcattle,andalthoughallthreeruminants(cows,goats

andsheep)sharemanysimilarities,theyarealsoverydifferentintermsoffeedingstrategiesand

thephysiologicalfunctionsthattheyprovide(milk,meatorwool)(Cannas,2019).Themost

importantdifferencebetweenthemisbodysizeandweightwhichareproportionallyrelatedto

theenergyrequirementsthattheywouldneed.Sincegoatsandsheeparesmallerthancows,they

have“highermaintenancerequirementsandalowerratiobetweenreticulorumenvolumeand

energyrequirementsthencattle”(Cannas,2019).Thisisbecausetheyhavelessrumenavailable

andsotheyneedtoeatmoretosatisfymaintenancerequirements.Additionally,goatsalsohave

moreselectivefeedingrequirementswhichwillcausethelowerrumentofill(Cannas,2019).

Sheepandgoatalsoneedtospendmoretimeeatingandchewingtheirfood–inorderforthe

foodtobegrounddowntotheproperparticlesizethatcanbepassedthroughtherumen(Cannas,

2019).Productionefficiencyreferstothemaximizationofobtainedproductoroutputs(suchas

milk)relatedtoinputs(suchasdrymatterintake,nutrientsandenergy).Inputsarelostwhen

thereareinefficienciesinthesystem–suchassurplusoffeedorextensiveenvironmental

impacts(Cannas,2019).Whenresidualfeedintakeisreduced,thereislessspendingonfeeding

costsandtheenvironmentalimpactoftheanimalsisalsoreduced(intheformofemissions

reductions)(Cannas,2019).Farmandanimalconditionscanaffectresidualfeedintake(breed,

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age,bodycomposition,digestion,etc)(Cannas,2019).Feedconversionsandoptimizingthe

amountoffeedusedforadequateconversiontomilkisimportantinreducingmethaneemissions

(Cannas,2019).Forlambsthiscanbeachievedbyfeedingapasturecomposedofrye-grasswith

increasinglevelsofdrymatterintake(Cannas,2019).Thiswouldincreasetheenergythatis

accessiblethroughgrainandleadtoareductioninemissions.Additionally,accountingforthe

amountofemissionseachanimalproducescanserveasaproxyfortheanimals’efficiency–less

gasindicatesgreatermetabolicefficiency(Cannas,2019).

b.Goatfeedingsystem

Indevelopingcountries–thegoatfeedingsystemsareoverallcharacterizedbylowinput

ofpoorqualityfeedsandpasturesandleadtooveralllowproductivity(Nampanzira,2015).This

isespeciallytrueincountriesthathaveaveryrainyanddryseasonwhichcanaffectthe

availabilityandqualityoffeed(Nampanzira,2015).Therefore,itisimportanttoimprovethe

feedingandnutritionoftheanimalsinordertoenhancegoatproductivity.Sincelandcanbe

scarce–thiscanlimittheamountofqualitypasturesavailableforthegoatsandthiswillcallfor

improvingfeedtechnologies(Nampanzira,2015).Oneexampleofthisisaddingleguminous

trees.Thiscanimprovedailyweightgainandthenutritionvaluesofthepasture,andultimately

increasetheproductivityoftheanimals(Nampanzira,2015).Feedavailabilityisalsoseasonally

dependant.Inthewetseason–goatsareabletohaveanabundanceoffeed(Nampanzira,2015).

However,inthedryseason–theirfeedneedstobesupplementedsincemostforagehasdriedout

andislackingincrudeprotein(Nampanzira,2015).

c.Sheepfeedingsystem

Inonestudydoneonhairsheep–grassesandforagesweresupplementedtoassess

whetherornotthiswouldimpacttheproductivityoftheanimals(Macías-Cruz,2009).Thiswas

doneduetotheseasonalandannualvariationsinforagequalityandamounts.Aforage

supplementswasfreshorangepulpderivedfromalocaljuicefactory(Macías-Cruz,2009).This

wasanidealalternativebecauseitwaslow-costandcouldreplacethetraditionalgrainand

forageinthelambs’diet.Thefreshorangepulpissimilartograinsandiseasilydigestedbythe

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hairsheep(Macías-Cruz,2009).Additionallyfreshjuicepulpreleasespectinwhichisacarbthat

canbefermentedintherumenandreleaseenergyforrapidmicrobialgrowth(Macías-Cruz,

2009).Itisalsoasolublefiber–whichmeansthatlesslactateisproduced,andthisleadstoa

healthierrumen.Unfortunately,citruspulpishighlyperishableandrequiresheavymonitoring

overtime(Macías-Cruz,2009).Thecitruspulpwasfoundtobereadilyfermentableandabetter

sourceoffiberthenthedryforagesbecausecitruspulphadahigherpectintolignincontent.The

FOP(freshorangepulp)alsoleadtoamorefatteningdietwhichwouldaidinweightgainofthe

animals(Macías-Cruz,2009).Additionally,theFOPalsostimulatedchewing–whichaidsin

ruminationandtheenergydensityofthedietoveralldecreasesasforagelevelsincrease

(Macías-Cruz,2009).Thedownsidetousingcitruswasthatitwouldneedtobecoupledwithan

otherforagesourcetoactivatethepositiveeffectsondigestion(Macías-Cruz,2009).Citrusalso

containsphenolsandtanninswhichcanreducepalatability-reinforcingthefactthattheFOP

needstobeusedinconjunctionwithanotherforagesource(Macías-Cruz,2009).

d.FeedingsystemsatGreenlandLivestockandResearchInstitute

AtGreenlandboththesheepandgoatsarefedthesameimportedfeedwhichislocally

mixed.Thefeedisalsoenrichedwithsoyaandcorninordertoincreasetheproteinandquality

ofthefeeds.Additionally,thegoatsareletouttopasturetograzethelocalgrassonadailybasis

andareabletoroamfreeaftermilkingforafewhoursadayastheyhaveextensivepastureland

tograzeon.Greenlandisalsolocatedinthenortheastpartoftheislandwhichisnotasbadly

affectedbydroughtconditionsassomeotherpartsoftheisland.Thispreferablelocationallows

forgrazingtobedoneduringmostpartsoftheyear.However,thepasturegrassesareinsufficient

asfeedisimportedtoprovideadditionalnutritionalsupporttotheanimals.Thesheeponthe

otherhandareexclusivelyfedtheimportedmixedfeedandonlygoouttopastureforbreeding.

Thismaybebecausethesheepareprimarilyraisedformeatconsumptionandthereforemay

needadditionalcaloricintakeandamorefatteningfeed.Thefeedforboththeanimalsisattimes

enrichedwithlocalsupplementssuchasorangejuicepulpinordertoincreasethequantityof

outputsfromtheanimals.

4.7MilksafetyandpasteurisationtechniquesinBarbados

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a. Milksafetystandards

Themicrobialstatusofrawmilkdeterminesitsqualityandchemicalcomposition

(Robinson,2008).Countsandtypesofmicroorganismsinfluencethekeepingqualityofraw

milkandtheefficacyofheattreatments.Milkdeliveredfromthefarmtoprocessingplants

containsmicrobesinvaryingamountsdeterminedbythehealthofcowornonbovinecattle,their

environment,feed,milkingequipment,watersupply,labour,coolingandtransportconditions

(Haenlein,2013).Themicrobialstatusofrawmilkintendedforhumanconsumptionis

determinedbylaw(EurpeanUnion,2006).BarbadosusestheEuropeanUnionsafetystandards

fordeterminingacceptablemilkforconsumption(Appendix1).Thisexperimenttoousesmilk

safetystandardsestablishedbytheEUinordertoassessmilksafetyofrawandpasteurisedGoat

andBlackbellySheepmilkattheGreenlandLivestockandResearchCentre.Moreover,amongst

theaforementionedcausesofmilkcontaminationinbovineandnonbovineruminants,amalistic

udderandmilkingequipmentarethemostserious,sincetheycanincreasethebacterialcountof

rawmilkbyseveralcfupermilliliter(Haenlein,2013).

b.Milkstudyofpathogens

Rawmilkisarichmediumforbacterialandpathogenicgrowththatcanleadtoseveral

foodbornediseases.Pathogenicdevelopmentoncheeseandmilkcanbehazardoustothehealth

ofconsumers,especiallytochildrenortheelderlypopulation(Foscino,2001).

MilkcontaminationcanoccurthroughmanypathwaysForone,itcanpassthroughthe

bloodduetoaninfectionintheudderorduringandafterthemilkingprocessduetopoor

handling(Gonzales-Barron,2017).Themainpathogenicbacteriafoundincow,sheep,andgoat’s

milkareBacilluscereus,Campylobacter,Staphylococcusaureus,E.coli,Salmonella,etc.

Althoughmanysheepandgoat’smilkundergoesfurtherprocessingtobeturnedintocheeseand

yoghurt,theheatstablepathogenscanwithstandtheseprocesses(Gonzales-Barron,2017).

InastudydonebyGonzales-Baron,theincidenceofL.monocytogenes,Salmonella,and

S.aureusingoatandsheepcheeseswasassessed.ResultsindicatedthatSalmonellahadthe

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lowestrateofcontaminationinbothsheepandgoatmilk,whileListeriahadahigherlevelof

contaminationinbothrawmilks.Itwasanalysedthattheroutesofcontaminationforboth

bacteriawerethroughpoorqualityfeeds,crosscontaminationoffeedandmanure,poorquality

housing,contaminatedmilkingequipmentandmastitis(Gonzales-Barron,2017).Additionally,

E.coliwasthesecondmostfrequentcontaminantwithahighprevalenceinrawsheepandgoat

milk.

Theequipmentusedindevelopingcountriesissometimesmorerudimentaryandless

sophisticated;thereismorehandmilking,longermilkingtimes,andpoormilkingfacilities

(Gonzales-Barron,2017).Additionally,cheesemanufacturedonthesamefarmwherethemilkis

producedcanbeexpectedtohaveahigherincidenceofpathogeniccontaminationandtherefore

wouldrequirestricterpreventativeandsterilizationpractices(Gonzales-Barron,2017).

InastudydoneinItaly,themicrobialcontentofrawgoatmilkintendedforcheese

makingwasassessed(Foscino,2001).Themainmicrobialgroups(Salmonella,Listeria,andE.

Coli)wereanalyzed.FoscinodidnotdetecteitherSalmonellaorListeriabuddiddetectone

incidenceofE.Coli.ThemostprevalentcontaminantdetectedwasS.aureus.Sinceheat

treatmentcanbedifficulttocarryout(duetoproductioncapacitiesandeconomiesofscale),

improvingthehygienicconditionsoftheanimals,milkingstages,andcheesemakingprocessis

essentialtocontrollingcontaminationandhazardousproducts(Foscino,2001).Themain

microorganismsthatwereregularlyfoundwereyeastandcoliforms(Foscino,2001)Theorigin

ofthefarmwasabigfactorindeterminingthemicrobialcompositionofgoatmilk(thisisdueto

differencesinfeeding,hygienicconditions,practicesonfarmandlocationofthefarm).The

amountofcoliformswasproportionallyrelatedtothehygienicpracticesdoneduringthe

productionandcoolingofthemilk(thosewithlittletonorefrigerationhadahighercoliform

count)(Foscino,2001).

c.waterasapossiblesourceofcontamination:

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Accesstocleanandsafedrinkingwaterisamajorhealthconcernformanydeveloping

regionsintheworld.Poorwaterqualitycancausemanygastro-intestinalandinfectiousdiseases

whenconsumed(Amenu,2016).Exposuretothesepathogenscanresultthroughtwochannels-

eitherdirectconsumptionofthewaterorindirectconsumptionofwaterwhenitisusedinthe

cleaning,processingandhandlingoffoodproducts-thisisespeciallycrucialordangerouswhen

non-potablewaterisusedintheproductionofrawmilkwhichisanexcellentmediaforbacterial

proliferation(Amenu,2016).

Hygienicpracticesareofutmostimportancewhenitcomestominimizingmicrobial

contamination(Amenu,2016)andensuringthequalityofthemilk.However,whena

contaminatedwatersourceisusedthesehygienicpracticescanbenulledoutifthewaterthatis

usedtocleantheequipmentandtheanimalsisnotsanitaryand/ornotregularlycheckedasa

sourceofpathogens.AsemphasizedbyKebedeAmenu’sstudy:“thewaterusedduringhandling

andprocessingofmilkproductscanbeapotentialsourceofmicrobialcontaminationwith

possiblenegativeconsequencestofoodsafety”.Thisindicatesthatthequalityofthewaterisjust

asimportantastheotherrawinputs-suchasthemilkinproduction.

Itisrecommendedthatthewaterthatisusedincleaningtheequipmentandmachinesand

intheprocessingofthefinaloutputs(themilk)shouldbeheldtoa“drinkable”standard-

thereforesafeforhumanconsumptionevenwhenitisjustusedforcleaningpurposes(Amenu,

2016).Thiscanposeachallengetomanysmallholderfarmerswhoareunabletoverifyorobtain

safeanddrinkablewaterthereforeincreasingthepathwaystocontaminationoftheirproducts

anddecreasingthefoodsafetyoftheirfinalproducts(Amenu,2016).

d.QualityassuranceofRawmilk:

GoodqualityGradeAmilkneedstobeabletofulfillcertainrequirementsputinplaceby

theEuropeanUnion’sIDF(internationaldairyregulations)guidelinesinordertomaintainthe

qualityofthemilkandtoinsurethatitissafeforsaleandconsumption(Park,2010).These

requirementsarethatthemilkissafetodrink;hasgoodflavor;isfreeofspoilagebacteriaand

somaticbodycells;anditscompositionisregulated(Park,2010).Therearecertainmethodsput

inplacesuchaspasteurization-aprocessthatisabletoremovepathogens-toensurethesafety

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ofthemilk(Park,2010).However,thiscanbeacostlyandtimeconsumingpracticeformost

smallfarmstoadopt.Therefore,sincethequalityofmilkishighlyimpactedbymanyfactorsthat

existonthefarm–thesefactors(feeding,handlingandmilkingpracticesforexample)needtobe

safeguarded,regulatedandcontrolledforbiosafetyduetotheirabilitytoaffectthequalityand

perishabilityofmilkdowntheline(Park,2010).

Moreover,goodqualitymilkneedstobefreeofpathogensormicroorganismswhichcan

damagethemilk–thisincludesforeignsubstancessuchasantibioticsandpesticideresidues

(Park,2010).Themainparameterswhichareroutinelymonitoredandregulatedarenutritious

constituentsinmilk;somaticcellcountandbacterialcount(whichislinkedtothehygienelevels

ofthefacility);pesticideresiduecontent;flavor,tasteandappearance(Park,2010).

Whenafarmisabletoproperlycareforitsanimalsandimplementgoodmanagement

practicessuchaspromotinghealthylactationandsanitarymilkingproceduresandequipment,

havingqualityfeedfortheanimalscanheavilyinfluencethetasteandqualityoftheproduct

(Park,2010).Goodqualitymilkbeginswithagoodbalanceddiet,goodcleaningoftheliving

andmilkingquartersoftheanimals,andproperventilationbeforeandduringmilking(Park,

2010).Workersshouldalsopracticepropercleaningpracticesofthemilkingequipmentand

routinelycheckforthecleanlinessandfunctionalityofthemilkingequipment(Park,2010).

Moreover,thefundamentaldifferencesinmilksecretionsystemsbetweendairyspecies

influencethepathogenicsusceptibilityofthemilktheyproduce.Goatssecretemilkthroughthe

apocrineprocesswhilecowsandsheepsecretemilkthroughthemerocrineprocess.This

contributestogoat’smilkhavingahighersomaticcellcountcomparedtocow’smilk.Yet,the

somaticcellcountregulationsformilkqualitystandardsareenforcedforbothspecies.Though

Goatmilkhasahighersomaticcellcountthancow’smilkduetotheapocrinesystemmilk

production,thisisn’tanindicatorforleukocytecount(indicatorofinfection)(Park,2010).Due

tothisproblem,goatproducershaveactivelypursuedtheproblemofSCClegalthresholds.

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Basedonthis,thisexperimentassumesahighercoliformcountinrawmilkforgoat’smilk,as

opposedtosheepmilk.

d.QualityassuranceatGreenland:

Greenlandtriestoimplementtheseprinciplesandpreventdiseaseintheirmilkbymaking

suretheiranimalsareingoodhealthandbycontinuouslycleaningeverythinginvolvedinthe

milkingprocess.Thegoats’uddersandfeetarecleanedwithiodinebeforemilking,andthepart

ofthemilkingmachinethatmimicsahandautomaticallycleansitselfaftereveryuse.An

alkalineacidispumpedthroughthefiltrationsystemeverydaytoremoveanyfatbuildupthat

mayhaveoccurredduringmilking.Everytwoweeks,themachineisgivenamorethorough

cleaningandistakenapartsothatindividualcontactsurfacescanbecleanedwithchlorineand

hydrochloricacid.Greenland’scurrentcleaningpracticeswereprescribedtothembyanoutside

companytheyconsulted.Themachinecamewithacleaningliquid,buttheemployeesat

Greenlanddidnotfindthatitworkedasitdidnotsuccessfullyremovefatsolidsorcalcium

buildupfromthepiping.Thecompanytheybroughtingavethemnewprocedurestofollow,and

thoseproceduresaretheircurrentprotocol.

4.8Treatmentofmilk

a. Coldstorageinfluencesonmicrobialcompositionofrawgoatmilk:

Standardsandspecificationsforpropermanagementofmilkarecrucialinavoiding

negativeeffectsonthemilk’squality(Yamazi,2013).Topreservethequalityofthemilk,it

shouldbecooledinarefrigeratoraftermilkinguntilconsumed.Despitetheslowedgrowthof

microorganismsthatcomeswithcooling,itisstilleasyforpsychotropicstocreatespoilage

enzymeswhichmaydirectlyimpactthequalityofthemilkanditsproducts(Yamazi,2013).The

enzymesproduceddamagemilkqualitybyinterferingwithcoagulationofmilkduringcheese

production.Thiscanbeavoidedwhencareofanimalsandpropermilkingpracticesare

safeguarded(Yamazi,2013).

b.Heattreatmentofmilk:

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Heatingmilkpromotesaggregationanddecreasesacidificationwhichallowsforthe

correctassociationsofproteinsinfermentedmilktoappear(BonfimdaSilva,2019).Milk

heatedto65°Candthenrapidlycooled(whichisthestandardprotocolforathomeboiling

pasteurization)causesatemporarydecreaseandthenanincreaseinmilkviscosity.Thisrenders

theproteinsofthemilkinsoluble(BonfimdaSilva,2019).However,thesechangestothe

textureprofileoccuronamicrolevelandareonlydetectableinmilkthatisexposedtohighheat

orsuperheated,suchascondensedmilk(BonfimdaSilva,2019).Thereisnonoticeablechange

oftexturewhensheepmilkisheated(totemperaturesof90-95°Cfor5-10minutes)sinceithas

ahightotalsolidscontent(BonfimdaSilva,2019).

Pasteurizationisacommonpracticeforkillingharmfulandpathogenicorganismsinraw

milk.Pasteurizationcanimproveshelflifeaswellastransportationeaseandtime(Bonfimda

Silva,2019).Controllingforcrosscontaminationbefore,during,andafterpasteurizationis

importantinensuringthefoodsafetyofthemilk(BonfimdaSilva,2019).However,goatmilkis

sensitivewhenitcomestoheattreatment,morespecificallytoUHT(ultra-hightemperature)

methodsofpasteurization(BonfimdaSilva,2019).Differentformsofpasteurizationhavebeen

recommendedfordairyfarmerssinceUHTcandestabilizeandchangethetasteprofileofthe

milk(BonfimdaSilva,2019).

Theimportanceoftheuseofheattotreatfoodhasbeenknownforalongtime-almost

allofhumanhistory,buttheprocessofmilkpasteurizationhasonlyrelativelyrecentlybeen

refinedandpopularized(Steele,2000).Heatwasusedinthepreservationofvinegarin1782by

Scheele,aSwedishscientist,butitwasn’tuntil1824thatProfessorWillamDeweesofthe

UniversityofPennsylvaniarecommendedfeedingbabieswithmilkthathadbeenheatedand

cooled(Steele,2000).However,thepasteurizationprocesslargelyinplacenowislargelythe

resultoftheworkofFrenchscientistLouisPasteur(Steele,2000).Histechniqueofparboiling,

orunderboiling,waspioneeredduringhiseffortstosolvetheproblemofwinespoilage(Steele,

2000).Thisparboilingtechniqueinvolvesheatingtheliquidtoalevelsufficientenoughtokill

thespoilageorganismswhileretainingthequalityoftheliquid.Pasteurperfectedthismethod

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withwinein1864,andhelaterappliedittobeer(Steele,2000).ThereisnoevidenceofPasteur

havingusedthismethodonmilk,buthisworksetthebasisforwhatmilkpasteurizationwould

become(Steele,2000).Inthelate19thcentury,NathanStrausadoptedtheissueofmilk

sterilizationasaphilanthropicmissionashewasdiscouragedbythenumberofinfantdeathshe

sawresultingfromtheconsumptionofrawmilk.Strausworkedtirelesslytopopularizethe

processofmilkpasteurizationboththroughouttheUnitedStatesandinEurope(Steele,2000).

Thereareafewdifferentmethodsofpasteurization.Flashpasteurizationinvolvesheating

milktotemperaturesbelowitsboilingpoint,usuallybetween70and80degreescelsius,forjust

asecondbeforecoolingitbackdown(Steele,2000).Thisformofpasteurizationwasoneofthe

earliestformsofcommercialpasteurization,andvatorholderpasteurization,whichinvolves

holdingmilkataslightlylowertemperature(60degreescelsius)foralongerperiodoftime,

followedasthemorepopularandeffectivemethodofcommercialpasteurization(Steele,2000).

Amethodofpasteurizationthatcanbedoneonthehouseholdleveliscookingorboiling:a

methodwhichpeoplehaveturnedtoforalongtime(Steele,2000).Inthisexperiment,emphasis

hasbeenmadeonhomepasteurisingi.e.stovetopheatingofmilkthroughadoubleboiling

methodwherethemilkisindirectlyheatedthroughheatconductionfromboilingwater(Steele,

2000).

Currently,noneofthemilkthatisproducedatGreenlandispasteurized.Thismeansthat

manypeoplewhobuyfromGreenlandconsumerawmilkorhomeboiledmilk.This

consumptionofrawmilkcanleadtodisease,andthereareriskstodrinkingboiledmilkifthe

boilingisnotdoneproperly.PinehillDairy,ownedbyinternationalbeverage-giantABInbev,

usesultra-highpasteurizationonallofthemilkthatpassesthroughitsfacility.Pinehillreceives

milkfromavarietyoflocalfarmers,teststhemilktomakesureitisuptoquality(notwatered

downandcontainsthecorrectamountofcream),andpasteurizesitbeforemakingitintooneof

theirsignatureproducts.Pinehillsuppliesalargeamountofpasteurizedmilktotheisland,but

thepresenceofunpasteurizedmilkinBarbadosisnotnegligibleasGreenlandandothersmall

farmsselltheirproductswithoutpasteurization.

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5.MaterialsandMethodology

InthisExperiment,threetypesofmilkweretested:i.blackbellysheepunfiltered,ii.goat

unfilteredandiii.goatfiltered.AsthereisnoequipmenttomilkblackbellysheepatGreenland

currently,thisexperimentlacksasheepfilteredtreatmentgroup.Milkfromtwotothreesheep

waspooledtogetherforsampling.GoatMilkwasderivedfrommarkedgoatsbelongingto

Seeaanen,TogenburgeandAlpinebreeds.Themilksweresampledonthreedifferentdaysovera

periodofthreeweeks.

Inthefirstround,rawgoatandsheepmilkwassampledfortotalcoliformsandSalmonella

spp.Duringthesecondround,alowerlevelpasteurisationwasconductedat45°Cfor10sand

15sinadditiontoanotherroundofrawmilktesting.Duringroundthree,ahigherlevel

pasteurisationwasexecutedat73.8°Cfor10sand15sinadditiontoathirdandfinalroundof

rawmilktesting.AnegativeandpositivecontrolfortotalcoliformsandSalmonellaspp.Was

executedoneachdayoftesting.

FigA:Avisualrepresentationoftheexperimentalprotocolfollowedduringthreeroundsoftestingmilksamples.Raw,lowerlevelpasteurisedandhigherlevelpasteurisedgoatfiltered,goatunfilteredandsheepunfilteredmilkweretestedfortotalcoliformcounts.Rawgoatfiltered,goatunfilteredandsheepunfilteredmilksamplesweretestedforthepresenceofSalmonellaateachround.

Totalcoliformcounts:

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5.1PreparingVioletRedBileAgar(VRBA)

ReagentsandApparatus:

Yeastextract-3.0g,Peptoneorgelysate-7.0g,NaCl-5.0g,BilesaltsorbilesaltsNo.

3-1.5g,Lactose-10.0g,Neutralred-0.03g,Crystalviolet-0.002g,Agar-15.0g,Distilled

water-1.0liter

Methodofpreparation:

Suspendingredientsindistilledwaterandletstandforafewmins.Mixthoroughlyand

adjusttopH7.4±0.2.Heatwithagitationandboilfor2min.Donotsterilize.Beforeuse,cool

to45°Canduseasaplatingmedium.Aftersolidification,addacoverlayerabovetheagarof

approximately3.0to4.0mltopreventsurfacegrowthandspreadingofcolonies.

5.2SolidMediummethod:coliforms

Preparevioletredbileagar(VRBA)according1.1.Coolto48°Cbeforeuse.Prepare,

homogenize,anddecimallydilutesamples.Place1mLofthemilksampleintopeptonebroth

usingafinnpipette.Securethejarshutandswirl30times.Labelthisasthe10-1dilution.Place

1mLfromthisjarintoanotherpeptonebrothjar.Closeit,swirlandlabelas10-2Carrythisout

until10-5forrawmilksamplesandtill10-3forheatedmilksamples.Then,Labelpetridishes

withthemilksampleandheatingtime.Transfertwo1mLaliquotsofeachdilutiontotwopetri

dishestocreatetwoduplicatesforeachsample.Afterthis,pour10mLVRBAtemperedto48°C

intoplates,swirlplatestomix,andletsolidify.Topreventsurfacegrowthandspreadingof

colonies,overlaywith5mLVRBA,andletsolidify.Aftersolidification,invertplatesand

incubate18-24hat32°C(2).Finally,examineplatesundermagnifyinglensandwith

illumination.Countpurple-redcoloniesthatare0.5mmorlargerindiameterandsurroundedby

zoneofprecipitatedbileacids.Platesshouldhave25-250colonies.Lastly,toconfirmthatthe

coloniesarecoliforms,pickatleast10representativecoloniesandtransfereachtoatubeof

BGLBbroth.Incubatetubesat35°C.Examineat24hand48hforgasproduction.

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Salmonellaspp.Test:

5.3PreparingmediaforSalmonellatesting

ReagentsandApparatus

Blenderandsterileblenderjars,Sterile,16oz(500ml)wide-mouth,screw-capjars,

sterile500mlErlenmeyerflasks,sterile250mlbeakers,sterileglassorpaperfunnelsof

appropriatesize,and,optionally,containersofappropriatecapacitytoaccommodatecomposited

samples,Sterile,bentglassorplasticspreaderrods,Balance,withweights;2000gcapacity,

sensitivityof0.1g,Balance,withweights;120gcapacity,sensitivityof5mg,Incubator,35±2

°C,Refrigeratedincubatororlaboratoryrefrigerator,4±2°C,Waterbath,49±1°C,Waterbath,

circulating,thermostatically-controlled,43±0.2°C,Waterbath,circulating,

thermostatically-controlled,42±0.2°C,Sterilespoonsorotherappropriateinstrumentsfor

transferringfoodsamples,Sterileculturedishes,15×100mm,glassorplastic,Sterilepipettes,1

ml,with0.01mlgraduations;5and10ml,with0.1mlgraduations,Inoculatingneedleand

inoculatingloop(about3mmidor105l),nichrome,platinum-iridium,chromelwire,orsterile

plastic,Steriletestorculturetubes,16×150mmand20×150mm;serologicaltubes,10×75

mmor13×100mm,Testorculturetuberacks,Vortexmixer,Sterileshears,largescissors,

scalpel,andforceps,Lamp(forobservingserologicalreactions),FisherorBunsenburner,pH

testpaper(pHrange6-8)withmaximumgraduationsof0.4pHunitspercolorchange,pHmeter,

Plasticbags,28×37cm,sterile,withresealabletape.(Items23-24areneededintheanalysisof

froglegsandrabbitcarcasses.),Plasticbeakers,4liter,autoclavable,forholdingplasticbag

duringshakingandincubation,Sponges,non-bactericidal(Nascocat#B01299WA),or

equivalent,Swabs,non-bactericidal,cotton-tipped.

5.4:preparingbufferedpeptonewater(BPW)

ReagentsandApparatus:

Peptone-10g,Sodiumchloride-5g,Disodiumphosphate-3.5g,Mono-potassiumPhosphate-

1.5g,Distilledwater-1L

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Methodofpreparation:

Autoclaveat121°Cfor15min.FinalpH7.2±0.2.

5.5IsolationofSalmonella:

Add25mLofmilksamplein225mLbufferedpeptonewater(aspreparedin1.4)usinga

50mLpipetteandpipettefiller.Tightenlidandgentlyshakeincubatedsample.Ifthesampleis

slightlycloudypostincubation,thismaybeanindicationofthepresenceofSalmonellaspp.

Regardless,thenextstepshouldbeexecuted.Then,transfer0.1mlmixtureto10ml

Rappaport-Vassiliadis(RV)mediumandanother1mlmixtureto10mltetrathionate(TT)broth.

Vortex.Aftervortexing,incubateRVmedium24±2hat42±0.2°C(circulating,

thermostaticallycontrolled,waterbath).IncubateTTbroth24±2hat35±2.0°C.Vortexand

streak3mmloopful(10µl)incubatedTTbrothonbismuthsulfite(BS)agar,xyloselysine

desoxycholate(XLD)agar,andHektoenenteric(HE)agar.PrepareBSplatesthedaybefore

streakingandstoreindarkatroomtemperatureuntilstreaked.Repeatwith3mmloopful(10µl)

ofRVmedium.Useinoculatinglooptostreakcoloniesofbacteriaontoeachquadrantofthepetri

dish.Ensuresterilitybyflamingtheinoculatingloopeachtime.Incubateplates24±2hat35°C.

Finally,examineplatesforpresenceofcoloniesthatmaybeSalmonella.

5.6:TypicalSalmonellacolonymorphology

Pick2ormorecoloniesofSalmonellafromeachselectiveagarplateafter24±2hincubation.

TypicalSalmonellacoloniesareasfollows:

IftypicalcoloniesarepresentontheBSagarafter24±2hincubation,thenpick2ormore

colonies.IrrespectiveofwhetherornotBSagarplatesarepickedat24±2h,reincubateBSagar

platesanadditional24±2h.After48±2hincubation,pick2ormoretypicalcolonies,if

present,fromtheBSagarplates,onlyifcoloniespickedfromtheBSagarplatesincubatedfor24

±2hgiveatypicalreactionsintriplesugarironagar(TSI)andlysineironagar(LIA)thatresult

inculturebeingdiscardedasnotbeingSalmonella.

6.Results

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SectionA:Coliforms

TheEUGuide’sStandardforSafeDrinkingmilk(seeAppendix)isusedasareference

fordeterminingacceptablelevelsofmicrobesinmilksamplesinthisexperiment.Thevalues

highlightedinFig1.1-1.3depictvalidmicrobialcountsbasedonthegeneralacceptablerangeof

countablecoloniesonastandardagarplatei.e.25-250formostbacteria(Peeler,J.T.,2001).

Fig1.1:depictingno.ofcoloniescountedperagarplate(n,n)forunheated,Raw(nopasteurisation)

samplesofFilteredGoat,UnfilteredGoatandUnfilteredBlackBellySheepmilksamples;Round1.

MilkCondition TypeofMilk No.ofcoloniesatnDilution

10-1 10-2 10-3 10-4 10-5

Raw,unheated

Filteredgoat 31,41 7,3 2,0 0,0 0,0

Unfilteredgoat 0,0 4,8 1,0 0,0 0,0

Unfilteredsheep 0,0 0,0 0,0 0,0 0,0

Fig1.2:depictingno.ofcoloniescountedperagarplate(n,n)forRawunheated,lowerlevel

pasteurisationat45°Cfor10s)andlowerlevelpasteurisationat45°Cfor15s)forfilteredgoat,unfiltered

goatandunfilteredBlackBellySheepmilksamples;Round2.

MilkCondition TypeofMilk No.ofcoloniesatnDilution

10-1 10-2 10-3 10-4 10-5

Raw,unheated

Filteredgoat 60,42 0,0 0,0 0,0 0,0

Unfilteredgoat 0,0 4,8 1,0 0,0 0,0

Unfilteredsheep 0,0 0,0 0,0 0,0 0,0

Lowerlevelpasteurisationat45°C

for10s)

Filteredgoat 61,39 0,0 0,0 N/A N/A

Unfilteredgoat 0,0 4,6 0,0 N/A N/A

Unfilteredsheep 0,0 0,0 0,0 N/A N/A

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Lowerlevelpasteurisationat45°C

for15s)

Filteredgoat 31,31 0,0 0,0 N/A N/A

Unfilteredgoat 0,0 7,1 0,0 N/A N/A

Unfilteredsheep 0,0 0,0 0,0 N/A N/A

Fig1.3:depictingno.ofcoloniescountedperagarplate(n,n)forrawunheated,higherlevel

pasteurisationat73.8°Cfor10s)andhigherlevelpasteurisationat73.8°Cfor15s)forfilteredgoat,

unfilteredgoatandunfilteredBlackBellySheepmilksamples;Round3.

MilkCondition TypeofMilk No.ofcoloniesatnDilution

10-1 10-2 10-3 10-4 10-5

Raw,unheated

Filteredgoat 114,101 11,15 1,2 0,0 0,0

Unfilteredgoat 0,0 0,0 0,0 0,0 0,0

Unfilteredsheep 0,0 0,0 0,0 0,0 0,0

Higherlevelpasteurisationat73.8°C

for10s)

Filteredgoat 0,0 0,0 0,0 N/A N/A

Unfilteredgoat 0,0 0,0 0,0 N/A N/A

Unfilteredsheep 0,0 0,0 0,0 N/A N/A

higherlevelpasteurisationat73.8°C

for15s)

Filteredgoat 0,0 0,0 0,0 N/A N/A

Unfilteredgoat 0,0 0,0 0,0 N/A N/A

Unfilteredsheep 1,1 0,0 0,0 N/A N/A

Fig1.4:depictingcoliformconfirmationtestsresultsforcoliformsonpetridishesthatshowedgrowthduringRound2andRound3:RawFilteredGoatmilkonMondayandTuesday,FilteredGoatMilkpasteurisedunderlowheatconditions(45°C,10s)andFilteredGoatMilkpasteurisedunderlowheatconditions(45°C,15s).Positive(+)isanalysedqualitativelybyacloudygreenappearanceandthepresenceofgas.Negative(-)isindicatedbyacleargreengaslesssolution.NocoliformconfirmationtestwasexecutedforRound1duetoaforementionedlimitations.

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Colonyn ColiformConfirmationTestResults

FilteredGoat,Raw(Round2)

FilteredGoat,Raw(Round3)

FilteredGoat,45°Cfor10s

Filteredgoat,45C°for15s

1 + + + +

2 + + + +

3 + + + +

4 + + + +

5 + + + +

6 + + + +

7 + + + +

8 + + + +

9 + + - +

10 + + - -

Percentageoftruecoliforms(%)

100% 100% 80% 90%

SamplecalculationsforEnumerationofColiforms:1.7.Samplesthattestedpositive(+)for100%truecoliformsasdepictedinFig1.4wereenumerated

usingstandardCFUcountmethod:

CFU=(no.ofcoloniesxdilutionfactor)/(volumeofcultureplate)

No.ofcolonies=averageofcolonies=(ncoloniesfromsample1+ ncoloniesfromsample2)/(2)

No.ofcolonies=(60+42)/(2)=51

E.g.forFilteredGoat,Raw(Round2):

CFU=(51x101)/1mL

=510colonies/mL

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1.8Samplesthattestedbelow100%fortruecoliformsasdepictedinFig1.4areenumeratedby

alternativelyi.e.including%confirmedcoliforms.

CFU=(no.ofcoloniesxdilutionx%confirmedcolonies)/(volumeofcultureplate)

E.g1.FilteredGoat,45Cfor10s(lowerheatpasteurisation):

CFU=(50x101x0.80)/(1mL)

=400colonies/mL

Fig1.5:depictingtrueCFUcountsforcoliformpositivesamplesi.e.FilteredGoat,Raw(Round1),

FilteredGoat,Raw(Round2),FilteredGoat,Raw(Round3),FilteredGoat,45°Cfor10s-lowerlevel

pasteurisationandFilteredGoat,45°Cfor15s-lowerlevelpasteurisation.

MilkTreatment ColonyFormingUnit(Colonies/mL)

FilteredGoat,Raw(Round1) 360

FilteredGoat,Raw(Round2) 510

FilteredGoat,Raw(Round3) 1075

FilteredGoat,45Cfor10s-lowerlevelpasteurisation

400

FilteredGoat,45Cfor15s-lowerlevelpasteurisation

279

SectionB:Salmonellaspp.

Accordingto1.6,whichdescribestheprotocoladheredtofordeterminingthepresenceof

Salmonellainmilksamples,ItwasdeterminedthatSalmonellawasabsentinallmilksamples

tested.Referto1c,Appendix.

7.Discussion TheresultsofbacterialdistributioninthesamplesarepresentedinSection1.6.Alltheraw

filteredgoat’smilkshadhighbacterialloadswhichaveragerangedfrom360-1000c.f.u./mlafter

incubation,plating,andenumerationusingtheCFUformulatoestablishnumberofcoliformsper

unitmilkduringthethreeroundsoftesting.Theterm“totalcoliforms”referstoawiderangeof

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gram-negative,rod-shapedbacteriathatsharesimilarfeatures.Thisincludesthermotolerant

coliforms,bacteriathatmaybeoffaecaloriginandsomethatmaybeisolatedfrom

environmentalsourcessuchasmilkingfilters.Asthemicrobialloadwashighestintheraw

filteredgoat’smilk,itisassumedthatenvironmentalcontaminantsthroughthefilter,

implements,handlingequipmentorthesourceofthewaterandchemicalstocleanandsterilise

thefiltrationsystemrespectivelymayhavepresentedanentrypathwayforpathogensintothe

milk.

Moreover,themicrobialloadpermLfoundinunheatedgoat’smilkvariedconsiderable

duringeachroundoftesting.Thecoliformcountinrawgoat’smilkatround2was41.7%thanit

wasatround1,andthecoliformcountinunheatedgoat’smilkatround3was199%higher

comparedtothecoliformcountinround2.Takenthat,basedontheresults,itisassumedthatthe

sourceofcontaminationatGreenlandisthefilter,theincreasingtrendofmicrobialloadover

threeconsecutiveroundsoftestingmayindicatethatthefilterwasnotbeingcleanedadequately,

leadingtoabuildupandproliferationofmicrobesinthefilterorbulktank.Theuncleanlinessof

thebulktankandfiltrationsystemmaybearesultofalackofcleaning,alackofthoroughness

ofcleaning,orthedifficultyofkeepingthingsbacteriafreeinthehotandhumidtropical

environmentofBarbados.

Itisalsonoteworthythatnotallcounteddataindicatingmicrobialpresenceinmilksamples

isrepresentativeoftruecoliforms.Thus,toconfirmthetruenumberofcoliformsacoliform

confirmationtestwasexecuted.10colonieswererandomlypickedfromcoliformcontaining

agarplatesandplacedintoindividualBGLBbrothcontainingtesttubes.Ifgaswasproduced

postincubation,thecolonyrepresentedatruecoliform.Thispercentageoftruecoliformsinthe

randomsampletestwasusedtoadjusttheformulausedtoenumeratethenumberofcoliforms

permLofmilk,asexecutedinFig1.5.Thus,thedataanalysedinthisexperimentisbasedon

truecoliforms.Additionally,somemilksamples,suchastherawfilteredgoatmilkatthe10-2

dilutioninroundone(Fig1.1),containthepresenceofmicrobesatcounts(i.e.7,3)thatfall

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belowtheacceptablecountablerangeof25-250(Peeler,J.T.,2001).Themathtoundothe

dilutionfactorputsthenumberofcoliformsaroundtheoneobtainedfromthe10-1

Dilution.However,the10-1dilutionisusedsinceonlyplateswithcountsbetween25and300are

used.

Thehypothesisofthisexperimentstatesthatthehigherlevelofpasteurisationat73.8°Cfor

10swouldformtheidealpasteurisationcondition,ensuringthatthemicrobialloadingoatand

sheepmilkwouldbebroughtdowntoalevelsafeforhumanconsumption.AsdepictedinFig

1.5,thereweremicrobespresentinmilksamplesafteritwasheatedatthelowerlevelof

pasteurizationat45°Cat10sand15swithcolonyformingunitsof400colonies/mLand279

colonies/mLrespectively.Fromtheseresults,itcanbeinferredthatthoughtherewasa30.25%

reductioninmicrobialloadwhentheheatingtimewasincreasedfrom10sto15s,thetotal

coliformcountateachtimewashigherthanthatpresentafterheatingthemilkatthehigher

temperaturei.e.73.8°C.

Moreover,whiletheemphasisofthisexperimentisbringingdownmicrobialloadinraw

milk,itisbasedontheassumptionthatallrawmilkcontainsacertainlevelofnaturally

occurringmicrobesduetothephysiologyofsmallruminantssuchasgoatsandblackbelly

sheep,thevariousgatewaysfortheentryofmicrobespresentedbytheprocessofmilking-

manuallyormechanically.Thus,theabsenceofanysignificantcoliformsinrawunfilteredgoat

andblackbellysheepmilkinallthreeroundsoftestingwaspeculiar.AccordingtotheMinistry

ofAgricultureFoodandVeterinaryServicesLab,itwasestablishedthatitisrareforrawmilk,

especiallyinthetropicalhumidenvironmentofBarbados,tobefreeofanymicrobes.Forone,

thismaybeduetotheprolificnatureofblackbellysheepandgoatatGreenland.AtGreenland,

goatandblackbellysheeprearingdonotinvolvetheinjectionofhormonestoregulate(increase

thefrequencyof)lactatingcyclesasthespeciesofsmallruminantsaresturdyandprolificinthe

Barbadiansetting.Antibioticresiduesinmilkarecausedbythedirecttreatmentofdairycattle

withantibioticsinfeed.AretrospectiveobservationalstudyconductedatWisconsindairyfarms

foundapositivecorrelationbetweenantibioticresiduesandhighsomaticcellcounts(SCC)in

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milk(P.L.Ruegg,T.J.Tabone,2000).Thismayformpartoftheexplanationforrawgoatand

sheepunfilteredmilkatGreenlandbeingvirtuallyfreeofanycoliforms.

Secondly,Greenland’smanualmilkingmeasuresmaybehygienicandposeminimal

gatewaysofopportunityforentryofmicrobes.AccordingtoGreenlandResearchandLivestock

Institute,themeasurestakentomaintainsterilityofthemanualmilkingsystemincludetheusage

ofgloves,regularchangingofgloves,andthesanitisingofmilkingcontainerswithchemicals.

Moreover,themilkisstoredatrefrigeratortemperatureof4°Crightaftermilkingtoprevent

thermophilicmicrobesfromproliferatingandcausingmilkspoilage.

AlloftheSalmonellatestscamebacknegativeindicatingtheabsenceofSalmonellain

Greenland’sgoatandsheepmilk.However,intheunfilteredrawgoalmilksamplefromJuly

16th,therewasblackgrowthonthepetridishthatwarrantedfurthertestingtodetermineifitwas

Salmonellaornot.Whentheblackgrowthwasplacedontriplesugarironagar(TSI)andlysine

ironagar(LIA)(whicharebothagarsthatallowforthegrowthofhydrogensulfideproducing

bacteria-ofwhichSalmonellaisone),someblackhydrogensulfidewasproduced.The

colorationoftheagarafterincubation,however,didnotchangeinawaythatindicatedthe

presenceofSalmonella(referto2aand2bintheappendix).Thisleavesopenthepossibilitythat

themilkcontainseitherahydrogensulfideproducingCirtobacterorE.colispecies.Further

testingshouldbeconductedonthemilkatGreenland,andspecificallyonthegoatsnumbered

9011,00922,and00889fromwhichtherawmilksamplesweretaken.

8.PotentialSourcesoferror

Tocollectthemilksamplesfortesting,thesampleshadtobetransportedfromGreenland

toaMinistryofAgriculturebuildinginSpeightstownwhereicewascollectedbeforebeing

transportedpastBridgetowntotheMinistryofAgriculture’sVeterinaryServicesLab.The

sampleswerenotoniceforthetenminutesittooktogetfromGreenlandtotheMinistryof

Agriculturebuilding.Whilethisisashorttimeforthesamplestonothavebeencooled,thistime

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gapincoolingcouldbeapotentialsourceoferrorinthatitallowedmorecoloniestogrowthan

hadthesamplesbeenrefrigeratedsinceimmediatelyaftermilking.

Wewerealsounabletoconductanumberofsamplesthatwouldhavemadestatistical

analysispossibleforthisexperiment.Wehadplannedonconductingthreesamplesaweekovera

seriesofthreeweekswhichwouldhavegivenusninetotalsamples.However,wewereonlyable

tocollectsevensamplessincewedidnotreceiveasufficientamountofmilkthefirstweekwe

wentintothelab.Additionally,wetestedalowerlevelofpasteurizationthesecondweekthat

resultedinusnothavingrepeatresultsfromthehigherlevelpasteurizationweconducted.We

wouldhavelikedtohavemadeupforthisbycontinuingourtrialsforanextendedperiodof

time,butunfortunatelywedidnothaveaccesstothelabafterthethirdweek.Thisresultsinour

databeingpreliminaryandinthepossibilitythatourcountsareoutliersandnotanormal

reactiontothelevelsofpasteurizationweconductedsincewehavenonothingtocompareour

trialsto.

AnotherfactortoconsideristhepotentialofthemilkcontainersatGreenlandto

contaminatethemilksamples.Themilksamplesweregiventousinemptiedandcleanedout

halfgallonmilkjugs.Uponarrivaltothelab,themilkhadtobepipettedoutofthemilkjugsinto

sterileglassestoavoidtouchingtherimofthemilkjugandanypotentialcontaminantsthere.

Despitethefactthattheseprecautionsweretaken,thereisstillthepossibilitythatmorethanjust

therimofthemilkjugscouldserveasasourceofcontaminationandthatbysimplyputtingthe

milkintothejugsthemilkwasbeingcontaminated.Thiswouldmeantheresultsdidnot

accuratelyreflectthelevelofbacteriainthemilkbutinthemilkandthejugaswell.

Anotherstudyresearchedtheeffectofboilingmilkonmicrobialcontentofcowand

buffalomilk.Theresearchwascarriedoutwithasimilargoaltoourwork–to“determinethe

properboilingperiodwhichresultsintherequiredsafetythroughapropershelflifewith

minimumqualitydeterioration”(Metwally,2011).Theexperimentlookedatcow’sand

buffaloes’milkwhileourslookedatgoatandsheep’smilk.However,theystillheatedthemilkat

100degC,80degCand90degCfor30seconds,1minute,5and10minutesandthencooledin

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anicebath.Theresultsofthisexperimentwasthattherewasasubsequentdecreaseinmicrobial

contentfromtheuntreatedmilktothesamplethatwasheatedat80degC,however,therewere

smallerimprovementswhenthemilkwasheatedforlonger.Boilingthemilk,broughtthe

lethalitylevelofthemilkdowntoacceptablegradeAqualitymilksuitablefordrinking.Thisis

similartoourfindings–wherethenumberofcoloniesdecreasedthemorethemilkwasheated.

Additionally,thecontaminatedmilkcamefromthebulktankmilkandnotfromthe

animalsthemselves.ResearchfromtheFAOindicatesthat“themilkcontactsurfacesofmilking

andcoolingequipmentareamainsourceofmilkcontaminationandfrequentlytheprincipal

causeofconsistentlyhighbacterialcounts.Simple,inexpensivecleaninganddisinfecting

routinescanvirtuallyeliminatethissourceofcontamination”(FAO).Thisindicatesthatthe

surfacesthatthemilkcomesintocontactwith–thebulktankorthemilkingequipmentisnot

gettingcleanedaseffectivelyasitshouldandthesourceoftheproblemistheequipmentitself

andnottheanimals.TheFAOalsoindicatesthatthe“waterforDairyusemustbeeitheran

approved,pipedsupplyorchlorinated(50ppm)beforeuse.Inhardwaterareas,milkingand

ancillaryequipmentmustbede-scaledperiodically”(FAO).Thissuggeststhatevenifthe

cleaningmethodsusedatGreenlandareeffective,theireffortscouldbeundoneiftheyareusing

acontaminatedwatersource.Thewaterthatisusedincleaningthemilkingequipmentwould

thereforeneedtobeheldatthesamewatersafetystandardsthatdrinkable/potablewaterisheld

at.Thisistoensurethatthewaterthatisbeingusedtocleanthefiltrationsystemandmilking

equipmentisnotleadingtoanincreasedmicrobialloadinthefinalproducts.Therefore,ifthe

cleaningprotocolsandchemicalsareproperlyused-however,contaminationpersistsitwouldbe

worthwhileforGreenlandtochecktheirwater(aswellaswaterstorageareas)forwaterborne

pathogens(E.coli)aswellastreatingthewatersource(throughasimilarhomepasteurization

technique)beforebeingused.Additionally,properhandlingofthewaterwouldalsoneedtobe

ensuredtosafeguardagainstincreasedintroductionofpathogensduetounhygienicpractices

implementedduringwatercollectionandstorage(Amenu,2016).

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9.Conclusion

ThisexperimenttaughtusmanythingsaboutBarbadosandaboutdesigningand

conductinganexperiment.Firstly,welearnedabouttheinformationgapbetweentheproducers

andconsumers-especiallyaroundpasteurizingmilk.Producersknowthatboilingthemilkisa

crucialsteptotakebeforeconsumption-however,manyconsumersareunawarethatthisis

importanttodoorthehealthrisksthatareassociatedwithrawmilkconsumption,whichis

becausemanyconsumersareunawarethatmilkistheperfectmediumforbacterialgrowthand

proliferation.Thereisalsoabiginformationgapinthepopulation.Mostelderlypeoplearemore

likelytodrinkmilkrawandfromthefarmerthentheyaretobuypasteurizedmilkatthestore.

ThiscouldbeduetogenerationalandculturaldifferencesaswellasalackoftrustintheUHT

pasteurizationprocess-believingthatwhenmilkispasteurizeditisconsidered“dead”milk.

Wealsolearnedaboutthestrongculturalpreferencesanddisliketowardssheepmilk,cheeseand

yogurtconsumption.ManyofthelabtechniciansattheVeterinaryServicesLabandtheworkers

atGreenlandhaveindicatedthatsheepmilkissomethingthateithertheypersonallywouldnever

consumenorwouldanyonetheyknow-especiallynottheelderly.However,someyoungpeople

maybemoreopentotryingit.

ConductinganexperimentinBarbadoshasbeenauniqueexperiencesincethisresearch

requiredalotofpatience,coordination,delaysandmisunderstanding.Giventhatwedidnothave

anestablishedtopicandhadtobuildtheexperimentourselves-weultimatelyneededtonavigate

ourwayaroundtheinfrastructureofBarbados-suchascontactingagovernmentlabtobeableto

testsamples-whichdidnotinformusthattheywouldbegoingonvacationtherebycutting

downlabtimeforus;equipmentmalfunctioningatGreenlandwithlittlerecommendationsor

helpfindingalternatives.Throughtheseinconvenienceswelearnedthatconductingexperiments

takestimeandpatiencetomakesureeverythingisgoingsmoothly.

SinceBarbadosisatropicalcountry-welearnedhowcrucialitistohaveproperpreservation

techniquesduetohotandhumidenvironment,andthereforeitisimportanttobeabletoreduce

contaminationsources.

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WerecommendthatthestaffatGreenlandtofigureoutwherethesourceofthe

contaminationforthemilkinthebulktankiscomingfrom-whetheritisfromananimalin

particular,aspecificelementoftheequipmentthatiscontaminated(thebulktank,ortheother

milkingequipment),orthestaff-improperlywashingtheirhandsandnotwearinggloves.

Additionally,untilthecoliformcountisreducedtoanacceptablelevel(lessthan100CFU

accordingtotheEU’sstandard),routinemicrobialtestingshouldbedoneinordertoensurethe

qualityofthemilkthattheyareselling.

WealsorecommendthatGreenlandprintlabelsontheirbottlesindicatingtheidealmilk

boilingtimeandtemperature(boilingthemilkinadoubleboileruntilitreaches78degrees

Celsius).Wealsorecommendthattheselabelsindicatethatboilingthemilkdoesnotimpactthe

qualityofitsnutrients.Additionally,thelabelshouldincludetheassociatedhealthriskswith

drinkingunpasteurizedmilksuchas(gastrointestinalandstomachproblemsaswellaslongterm

risksandcomplicationswhichcanbeespeciallyharmfultosensitivegroupssuchastheelderly,

youngchildrenandpregnant/breastfeedingwomen).

AnotherrecommendationsforthestaffatGreenlandistoinvestigatenewcleaning

protocols.Themanufacturerrecommendedprotocolswereconsidereddefectivewhichleadto

theimplementationofnewones;however,thesenewprotocolsareeithernotfollowedproperly

ortheyhavealsobecomedefective.Therefore,werecommendthatGreenlandimplementand

enforceaneasytofollowstandardoperatingprotocolthatthestaffcanfollowonaweeklyand

dailybasistoensurethatthemachinesareactuallycleaned.Webelievethatthemachinesat

Greenlandarenotproperlytakencareof.Thelacticheckmachine–whichisusedeverythree

monthstochecktheproteincontentofthemilk-wasdefectivewhenwewenttosampleour

milksamplesfortotalfatandproteincontentofthegoatandsheepmilk.Additionally,residue

andbuildupcameoutofthemachinewhenattemptstocleanitweremademakingitappearasif

themilkisnotregularlycheckedpertheirrecommendations(every3months).Sincethis

machinewasnotwelltakencareof–itisfairtowonderwhethertherestoftheirmachinesare

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properlycleanedonabi-weeklybasisasdescribed.Apropercleaningscheduleandoversightis

acrucialsteptoensuringthequalityofthemilk.

Futureexperimentswouldideallybecenteredaroundtestingthenutritivecontentofthe

milksbeforeandafterboiling-suchasthefatandproteincontentandcomposition.Ifissues

withmilkqualitypersist,testingdifferentcleaningproductsormethodsthateffectivelyreduce

contaminationcanalsobedone.Additionally,thewaterthatGeenlandusescouldbelookedatas

apossiblecontaminationsource,andsocouldthetreatmentoftheanimalsthere.Additionally,

sinceBlackbellysheepmilkisnotaculturallypopulardrink-itcouldbeinterestingtofinda

waytopromoteitanditshealthbenefitsamongstlocalpopulationsoramongstthetourism

sectors.

10.Acknowledgements

WewouldliketothankJohnVaughnandDebraRollinsoftheGreenlandLivestock

ResearchStationoftheMinistryofAgriculture,Food,FisheriesandWaterResource

Managementforhelpingusconceptualizethisprojectandforprovidinguswithmilksamples

andculturalinformationaboutmilkinBarbados.WewouldliketothankCoreyLowe,adairy

technicianatGreenland,whocollectedallofthemilksamplesandwalkedusthroughthe

proceduresGreenlandusestocleanitsequipment.WealsoextendabigthankyoutoDavid

Elcock,PamelaWhitehall,KerrilynPilgrim,andMaxineMorrisoftheMinistryofAgriculture’s

VeterinaryServicesLaboratoryforallowingustousetheirlaboratorytoconductourcoliform

andsalmonellatestingandforgivingustheknowledgeofhowtocarryoutthoseprocedures.

ThankyoutoDavidElcock,thecoordinatoroftheVeterinaryServicesLab,forallowingusto

usethefacilityandforcoordinatingtherequiredpermissionforustoworkthere.

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11.Appendix1a.VRBAenrichmentmediashowinggrowthofcoliformcoloniesinagar.Coloniesare

characterisedbypinkregularcirclessurroundedbyathinfuzzypinkhalo.

1b.Petridishesgroupedaccordingtodilutionandmilkconditionbeforecountingcoloniesof

coliforms.

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1c.PetridishwithRappaport-Vassiliadis(RV)andtetrathionate(TT)mediatotestforthe

presenceofsalmonella.Onthetopleftquadrant,ablackstriatedcolonyindicatedthat

Salmonellaspp.maybepresentintherawgoatunfilteredsamplefromRound3oftesting.The

FoodandVetserviceslabexecutedanothertesttoconfirmifthecolonieswereSalmonella.

Resultswerenegative;theymay,however,indicatethepresenceofaCitrobacterspeciesofan

H2SproducingE.coli.Additionally,coliformsgrewontheseplatesaswell.

2a.AnegativesalmonellareactionfortherawgoatunfilteredsampletakenonJuly16th2019.

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2b. Thepositivecontrolreactionandcolorationof

Salmonella-notablydifferentfromthe

negativereactionaboveinthatthepurpletube

hasnoyellowbuttlikethetubeaboveandin

thatthereispinkinthesecondtubewhilethere

isnopinkinthesametubeabove.

TabledepictingtheEUstandardlevelsof

acceptablebacterialprofilesusedbythe

BarbadosMinistryofAgriculture’s

VeterinaryServicesLaboratory

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