commiss decis februari lai method analysi test raw milk heat treat milk eec commiss european commun regard treati establish european econom commun regard council direct eec august health anim health problem affect intra commun trade heat treat milk amend direct eec articl thereof articl commiss lai method analysi test monitor complianc standard relat raw milk heat treat milk raw milk lai method determin plate count cell count freez point presenc antibiot pasteur milk lai method determin absenc pathogen number coliform plate count absenc phosphatas presenc peroxydas absenc antibiot freez point steril milk uht ultra high temperatur milk lai method determin plate count absenc antibiot technic reason opportun step lai refer method analysi test ensur complianc standard continu examin condit routin analyt test method appli await result examin respons member state routin method ensur complianc standard laid direct eec determin refer method analysi test includ determin analyt procedur lai precis criteria ensur uniform interpret result measur provid decis accord opinion stand veterinari committe adopt decis articl refer procedur analysi test raw milk determin freez point enumer micro organ plate count test oc enumer somat cell detect antibiot sulphonamid articl refer procedur analysi test pasteur milk determin freez point determin phosphatas activ determin peroxidas activ enumer micro organ plate count test oc enumer micro organ plate count test oc enumer coliform coloni count oc detect antibiot sulphonamid detect pathogen micro organ articl refer procedur analysi test uht milk steril milk enumer micro organ plate count test oc determin freez point detect antibiot sulphonamid articl refer procedur analysi test precis criteria appli effect collect sampl carri accord rule set annex articl refer procedur analysi test refer articl set annex articl decis reexamin decemb order account develop scientif technic knowledg articl decis address member state brussel februari commiss rai mac sharri member commiss oj oj annex content page ii gener provis ii sampl raw heat treat milk gener provis introduct gener provis respect reagent equip express result precis test report compet author member state enforc laboratori charg sampl test milk respect gener provis reagent water mention made water solut dilut rins purpos distil water deioniz water deminer water equival puriti microbiolog purpos free substanc affect influenc growth micro organ test condit refer made solut dilut indic solut water dilut water meant chemic chemic recogn analyt reagent qualiti equip list equip list equip refer procedur item special item specif analyt balanc analyt balanc mean balanc capabl weigh mg express result result result state analyt report arithmet obtain test satisfi repeat criterion state method repeat criterion satisfi test repeat result declar invalid calcul percentag result calcul percentag mass sampl precis criteria repeat reproduc precis criteria procedur defin repeat absolut differ singl test result obtain procedur ident test materi condit oper apparatu laboratori short interv time li reproduc absolut differ singl test result obtain procedur ident test materi condit oper apparatu laboratori time li valu repeat reproduc criteria relev claus procedur repres confid level interv defin iso edit calcul result recogn collabor trial evalu procedur procedur collabor trial carri case valu repeat reproduc estim collabor trial studi refer plan conduct accord intern guidelin test report test report method analysi result obtain addit give detail procedur method analysi option circumst influenc result obtain test report give inform complet identif sampl ii sampl raw heat treat milk scope field applic procedur specifi refer procedur sampl raw heat treat milk procedur sampl transport storag sampl applic raw milk produc deliveri raw heat treat milk storag transport tank procedur applic sampl heat treat milk direct consumpt gener sampl raw heat treat milk can tank carri skill oper suitabl train undertak sampl milk consid compet author test laboratori instruct sampl personnel sampl techniqu ensur sampl repres conform entir batch consid compet author test laboratori instruct sampl personnel mark sampl ensur unambigu ident sampl sampl equip gener sampl equip made stainless steel suitabl materi adequ strength construct suitabl intend purpos mix sampl plunger agit mix liquid contain suffici area produc adequ mix product caus develop rancid flavour dipper solid handl suffici length enabl sampl depth contain obtain capac dipper ml sampl contain closur glass suitabl metal plastic materi sampl equip includ contain closur construct chang sampl affect result examin surfac sampl equip sampl contain clean dry smooth free crevic corner round sampl equip microbiolog examin sampl equip includ contain addit provis steril sampl equip success sampl clean steril sampl accord instruct requir laid test laboratori compet author integr success sampl preserv sampl techniqu gener irrespect test perform milk mix prior sampl manual mechan mean sampl immedi mix milk agit sampl milk tank time test sampl microbiolog examin volum sampl test requir capac sampl contain fill complet sampl allow proper mix content test avoid churn transport manual sampl sampl milk bucket can obtain agit rapidli move plunger bucket ensur milk properli mix cream adher neck sampl repres consign obtain accord sampl refriger farm milk tank vat mechan manual agit milk suffici homogen obtain volum milk mechan agit mix milk carri manual agit sampl weighbowl essenti milk tip weighbowl adequ mix addit manual mechan agit ensur distribut fat volum consign sampl exce capac weighbowl sampl repres consign obtain accord sampl divid bulk quantiti milk sampl contain repres quantiti contain note quantiti milk sampl relat sampl contain test individu mix portion respres quantiti amount proport quantiti contain sampl sampl bulk proportion amount mix sampl larg vessel storag rail road tank mix milk procedur sampl mix content larg vessel storag road rail tank mechan agit advis extent mix period time milk rest effici procedur mix appli circumst demonstr adequ purpos analysi envisag criterion mix effici influenc similar analyt result sampl part consign outlet tank interv discharg procedur mix milk consid effici differ fat content sampl condit larg vessel bottom discharg outlet discharg point small quantiti milk repres content mix sampl prefer manhol sampl discharg outlet run suffici milk ensur sampl repres mix content larg vessel storag rail road tank carri mechan agit built tank driven electr motor propellor agit driven electr motor manhol agit suspend milk case rail road tanker recircul milk transfer hose attach tanker unload pump insert manhol clean filter compress air case minim air pressur volum prevent develop rancid flavour automat semi automat sampl automat semi automat sampl devic sampl raw milk produc deliveri accord instruct test laboratori compet author equip prior regular interv undergo test prescrib respons author suitabl sampl procedur verifi order establish minimum volum milk collect properli sampl rate carri relat minimum volum sampl abil give repres sampl bulk proper agit automat semi automat sampl equip respons compet nation author prescrib minimum volum milk sampl minimum volum sampl maximum carri analysi perform precaut sampl heat treat milk direct consumpt retail pack sampl heat treat milk direct consumpt retail packag complet seal packag sampl packag machin cold room treatment establish process pasteur milk dai process sampl type heat treat milk pasteur uht treat steril number examin made accord instruct laid test laboratori compet author identif sampl sampl mark identif code readili identifi instruct test laboratori compet author ensur ident sampl transport storag sampl instruct condit transport storag time sampl analysi milk prepar test laboratori type milk procedur analysi instruct laid accord compet nation author instruct point includ transport storag precaut prevent exposur contamin odour direct sunlight contain sampl transpar store dark place sampl raw milk microbiolog analysi transport store oc oc time sampl analysi short event hour compet author accept storag temperatur oc oc time sampl analysi hour sampl pasteur milk microbiolog analysi transport store oc oc time sampl analysi short event hour milk sampl raw milk pasteur milk microbiolog analysi store laboratori refriger time sampl analysi short special precaut analys procedur annex ii content page determin freez point ii determin phosphatas activ iii determin peroxidas activ iv enumer micro organ plate count test oc enumer micro organ plate count test oc vi enumer coliform coloni count oc vii enumer somat cell viii detect antibiot sulphonamid ix detect pathogen micro organ determin freez point scope field applic procedur specifi refer procedur determin freez point raw pasteur uht treat steril milk partli skim milk skim milk apparatu thermistor cryoscop thermostat control bath cool electr refriger thermistor probe replac mercuri glass thermomet type instrument instrument seek maximum freez point plateau freez curv commerci reason set read fix time onset freez freez point curv differ milk milk standard solut calibr refer procedur requir plateau seek instrument fix time instrument routin screen measur freez point estim proport extran water milk provid acid sampl exce lactic acid ml definit freez point milk obtain measur procedur express degre celsiu oc principl test portion milk supercool temperatur depend instrument crystal induc mechan vibrat temperatur rise quickli plateau correspond freez sampl instrument calibr adjust give correct read standard solut procedur sampl milk condit plateau freez point milk degre celsiu apparatu glasswar usual laboratori equip cryoscop cryoscop consist thermostat control cool bath thermistor probe semi conductor resist thermomet circuit galvanomet read sampl agit devic initi freez sampl tube cool bath type cool bath immers type insul bath suitabl cool liquid stir temperatur differ point liquid exce oc temperatur liquid fluctuat oc nomin state manufactur import liquid cool bath maintain constant level surfac sampl tube volum mark cover cool liquid circul type continu stream suitabl cool liquid circul sampl tube temperatur liquid fluctuat oc nomin state manufactur suitabl cool liquid aqueou solut ethan diol ethylen glycol thermistor accompani circuit thermistor glass probe type diamet greater mm lead diamet greater mm time constant thermistor second note high work voltag current dissip constant thermistor temperatur rais oc surround oc maximum toler resist probe work posit cryoscop tip glass bead lie axi sampl tube point mm top tube figur page templat provid enabl user set probe posit note defin resist temperatur characterist thermistor formula temperatur kelvin resist ohm temperatur temperatur coeffici temperatur coeffici constant depend materi make thermistor current practic excess recommend measur read devic principl measur instrument oper principl seek plateau freez point curv plateau part curv temperatur remain constant oc minimum second manual oper resist thermistor balanc mean wheatston bridg similar devic highest qualiti stabl resistor toler greater temperatur coeffici exce amp oc variabl balanc resistor depart linear rang maximum mean adjust resistor calibr purpos measur dial graduat interv greater oc automat oper read devic provid discrimin oc rang oc stabil read devic circuit success indic temperatur vari oc linear circuit error greater oc introduc point rang oc oc instrument correctli oper stir wire wire metal inert milk diamet mm stir test portion stir wire adjust amplitud mount vertic lower end level tip thermistor probe toler mm posit permit stir wire vibrat later suffici amplitud state manufactur mm ensur temperatur test portion remain uniform determin time normal stir oper stir wire strike thermistor probe wall tube devic initi freez devic oper instantan initi freez sampl temperatur test portion rise freez point stir wire purpos procedur increas amplitud vibrat second stir wire strike wall sampl tube sampl tube sampl tube figur made glass mm long mm extern diamet mm intern diamet wall thick tube vari mm tube carri volum mark mm end mm base tube sampl volum ml electr suppli suppli voltag stabil apparatu extern fluctuat exce nomin main suppli fluctuat analyt balanc mark volumetr flask ml capac class dry oven ventil capabl maintain temperatur oc electr furnac ventil capabl maintain temperatur oc dessic reagent borosil glass distil water boil cool oc flask fit carbon dioxid absorpt tube sodium chlorid analyt reagent qualiti fine ground dri hour oc furnac altern dri oven oc hour cool room temperatur effici desicc prepar standard solut weigh amount tabl dry sodium chlorid weigh bottl dissolv distil water transfer quantit ml mark volumetr flask dilut mark water oc store oc stopper polyethylen bottl capac greater ml longer month freez point sodium chlorid solut oc nacl oc standard solut gentli invert rotat bottl time mix content time standard solut agit violent incorpor air sampl standard solut withdrawn bottl pour solut clean dry beaker pipett purpos solut bottl quarter full preserv fungicid thiomers solut older month calibr thermistor cryoscop cryoscop establish temperatur surround air deviat oc temperatur calibr carri cryoscop expos sunlight draught room temperatur oc ensur cryoscop good work condit accord manufactur instruct switch hour prior calibr check posit probe amplitud vibrat stir wire temperatur cool liquid select standard solut tabl close bracket expect freez point milk sampl test differ freez point solut prefer oc design cryoscop circuit thermistor design balanc specif freez point measur rang instrument case standard solut freez point calibr solut facilit calibr procedur manufactur pipett ml standard clean dry sampl tube oper cryoscop note sampl tube calibr made type glass wash rins deminer water time test milk sampl temperatur standard solut similar milk sampl adjust calibr control manufactur cryoscop read equal freez point standard solut repeat procedur standard solut continu altern success read solut adjust calibr control give correct freez point cryoscop readi directli freez point milk sampl applic correct prepar test sampl store sampl temperatur oc remov visibl foreign bodi solid butterfat sampl filter clean dry vessel mix sampl gentli filter inert milk effect laboratori temperatur milk test storag temperatur oc allow reach laboratori temperatur immedi commenc test standard solut milk sampl temperatur determin titrat acid milk time freez point test sampl acid exceed lactic acid ml milk examin uht treat steril milk stand minut open contain examin procedur preliminari check check level temperatur cool liquid accord manufactur instruct thermistor probe empti sampl tube sampl switch cryoscop ensur cool liquid properli stir circul cryoscop switch hour check temperatur cool liquid posit amplitud vibrat stir wire routin calibr check test run measur freez point standard sodium chlorid solut solut freez point oc consecut determin differ oc valu differ freez point standard solut oc recalibr cryoscop cryoscop continu carri routin calibr check cryoscop continu carri routin calibr check hour instruct manusfactur account determin freez point milk gentli invert rotat milk sampl bottl time mix content time sampl agit violent incorpor air pipett ml milk clean dry sampl tube remov excess pipett ensur probe stir wire clean dry wipe carefulli soft clean lintless tissu upward insert sampl tube calibr cryoscop manufactur instruct milk cool freez initi temperatur manufactur oc automat instrument temperatur observ digit read manual instrument precis achiev ensur freez start galvanomet pointer hair line coincid mark reason freez initi temperatur rang abandon test repeat test portion milk repeat sampl freez temperatur portion sampl warm oc held minut melt crystallin fat cool test temperatur test immedi temperatur milk initi freez rise rapidli remain virtual constant time fall freez point correspond highest temperatur reach period record note time temperatur remain constant time interv initi freez attain highest temperatur differ sampl sampl consider shorter water standard sodium chlorid solut milk essenti highest temperatur record measur satisfactorili complet remov tube rins water dry thermistor probe stir soft clean lintless tissu upward make duplic determin portion milk sampl differ freez point obtain greater repeat oc carri duplic determin portion sampl provid determin agre oc valu record calcul final result cool probe instrument place empti sampl tube sampl lower oper head order probe cool design cryoscop case essenti ensur probe adequ cool take measur make dummi determin consist read obtain express result calcul routin calibr check calibr confirm calcul accept duplic freez point valu obtain round decim place sum accept duplic valu odd number round nearest shown freez point oc duplic valu precis repeat oc reproduc oc ii determin phosphatas activ scope field applic procedur specifi refer procedur determin phosphatas activ pasteur milk definit phosphatas activ measur quantiti activ alkalin phosphatas present product express quantiti phenol microgram liber condit mention procedur ml pasteur milk milk phosphatas activ mg ml consid phosphatas neg principl phosphatas activ evalu amount phenol liber disodium phenyl phosphat ad sampl phenol liber react dibromoquinonechlorimid produc dibromoindophenol bluish colour measur colorimetr nm comparison made sampl phosphatas enzym destroi reagent barium borat hydroxid buffer dissolv barium hydroxid ba water make ml dissolv boric acid bo water make ml warm ml solut oc mix solut stir cool rapidli oc adjust ph addit solut filter store solut tightli stopper contain dilut solut equal volum water colour develop buffer dissolv sodium metabor nabo nabo sodium chlorid nacl water make ml colour dilut buffer dilut ml colour develop buffer ml water buffer substrat dissolv disodium phenylphosph dehydr phenol free ml buffer dissolv disodium phenylphosph ml colour develop buffer add drop bqc solut stand room temperatur minut extract colour form ml butan ol stand butan ol separ remov butan ol discard repeat extract solut store refriger dai develop colour extract prepar buffer substrat immedi dilut ml solut ml barium borat hydroxid buffer zinc copper precipit dissolv zinc sulphat znso copper ii sulphat cuso water make ml dibromoquinonechlorimid solut bqc solut dissolv mg dibromoquinonechlorimid bqc br cino ml ethanol store dark colour bottl refriger discard discolour month copper ii sulphat solut dissolv copper ii sulphat cuso water make ml phenol standard solut weigh mg pure anhydr phenol transfer ml volumetr flask add water mix make mark stock solut remain stabl month refriger dilut ml stock solut ml water mix ml mg phenol apparatu glasswar note glasswar stopper sampl tool carefulli clean recommend rins freshli boil distil water steam type plastic stopper phenol contamin permit usual laboratori equip analyt balanc water bath capabl maintain oc spectrophotomet suitabl read wavelength nm test tube mm mm prefer graduat ml pipett glass funnel conveni size cm diamet fold filter cm diamet medium filtrat speed volumetr flask prepar standard solut procedur note avoid influenc direct sunlight determin contamin trace saliva perspir give fals posit result avoid respect special attent pipet procedur prepar test sampl carri analysi directli sampl sampl refriger dai test portion pipett test tube ml test sampl tube control blank determin heat blank minut boil water cover test tube beaker boil water aluminium foil ensur entir tube heat cool rapidli room temperatur treat blank test sampl similar manner rest procedur add ml buffer substrat mix immedi incub sampl water bath minut mix content occasion time heat boil water minut manner cool rapidli room temperatur add ml zinc copper precipit tube mix filter dry filter paper discard ml refilt filtrat complet clear collect ml test tube add ml colour develop buffer add ml bqc solut mix colour develop minut room temperatur measur absorb control blank spectrophotomet wavelength nm repeat determin dilut sampl absorb measur exce absorb standard mg phenol tube measur prepar dilut mix volum test sampl volum part test sampl heat carefulli boil order inactiv phosphatas prepar calibr curv prepar suitabl rang dilut standard start standard phenol control blank mg phenol millilitr pipett ml water ml phenol standard solut test tube add test tube ml copper ii sulphat solut ml colour dilut buffer ml water ml bqc solut mix colour develop minut room temperatur measur absorb control blank spectrophotomet wavelength nm valu absorb obtain quantiti phenol ad calcul regress line procedur squar express result calcul formula calcul absorb read quantiti phenol regress line obtain calcul phosphatas activ express microgram phenol millilitr pasteur milk formula phosphatas activ quantiti phenol microgram obtain dilut factor dilut case dilut factor dilut factor ml test sampl connect precis repeat mg phenol ml reproduc preliminari mg phenol ml dilut appli limit mention refer result obtain dilut sampl iii determin peroxidas activ scope field applic procedur specifi refer procedur determin presenc absenc peroxidas enzym milk control pasteur definit peroxidas posit reaction milk properli pasteur blue colour occur second mix peroxidas neg reaction colour occur second mix principl peroxidas enzym decompos hydrogen peroxid atom oxygen liber oxid colourless phenylenediamin purpl indophenol storch test colour intens proport enzym concentr reagent solut phenylenediamin dissolv phenylenediamin warm oc water make volum ml solut dark brown bottl glass stopper store cool dark place prepar solut phenylenediamin form sediment dai discard solut hydrogen peroxid dilut ml hydrogen peroxid water make volum ml stabil add ml concentr sulphur acid litr solut hydrogen peroxid solut stabl month cool dark place bottl glass stopper prevent contact organ compound procedur introduc ml milk sampl clean test tube suitabl closur add ml phenylenediamin solut add drop hydrogen peroxid solut watch colour product second mix blue colour occur second addit reagent reaction unspecif iv enumer micro organ plate count test oc scope field applic procedur specifi refer procedur enumer micro organ mean coloni count techniqu oc procedur applic raw milk pasteur milk uht treat steril milk preincub oc dai definit design micro organ mean organ form countabl coloni incub aerob condit principl defin volum milk sampl mix cultur medium petri dish incub oc hour coloni count number micro organ ml raw pasteur milk ml preincub uht treat steril milk calcul apparatu glasswar usual laboratori equip apparatu hot air oven capabl oper oc autoclav capabl oper oc incub capabl maintain temperatur oc point ph meter temperatur compens accur ph unit water bath capabl oper oc len magnif len magnif talli counter mixer capabl mix ml milk sampl decim dilut ml diluent work principl eccentr rotat content test tube glasswar test tube suitabl closur suffici capac adequ head space mix ml primari dilut decim dilut flask capac ml tube ml capac hold cultur medium pipett plug cotton wool glass steril synthet materi unbroken tip nomin capac ml outlet diamet mm petri dish clear uncolour glass steril synthet materi bottom dish intern diamet mm intern depth mm minimum bottom irregular interfer count coloni steril glasswar glasswar steril procedur maintain oc hour hot air oven maintain oc minut autoclav autoclav care ensur adequ penetr steam equip steril contain tightli close flask loos lid glasswar steril autoclav dri vent steam pipett steril hot air oven cultur medium milk plate count agar composit yeast extract trypton glucos dextros skim milk powder agar depend gell properti agar water ml skim milk powder free inhibitori substanc verifi compar test skim milk powder free inhibitori substanc prepar suspend dissolv compon order yeast extract trypton glucos final skim milk powder water heat suspens assist procedur add agar heat boil stir continu agar complet dissolv steam minut filter filter paper check ph ph meter adjust ph steril oc solut mol sodium hydroxid hydrochlor acid distribut steril storag cultur medium dispens medium quantiti ml flask ml tube stopper flask tube steril autoclav oc minut check ph medium medium immedi store dark temperatur oc longer month prepar commerci dehydr cultur medium cultur medium prepar commerci dehydr medium follow manufactur instruct add skim milk powder dissolv compon adjust ph oc dispens steril store medium diluent pepton salin solut composit pepton sodium chlorid nacl water ml prepar dissolv compon water heat check ph ph meter adjust ph steril oc solut mol sodium hydroxid hydrochlor acid distribut steril storag diluent dispens diluent test tube quantiti steril tube ml diluent stopper tube steril autoclav oc minut check ph diluent diluent immedi store dark temperatur oc longer month prepar commerci dehydr diluent diluent prepar commerci dehydr tablet powder follow manufactur instruct adjust ph dispens steril store diluent procedur melt medium begin microbiolog examin melt requir amount medium quickli temper medium oc water bath prepar milk sampl mix milk sampl micro organ distribut evenli rapidli invert milk sampl contain time foam avoid foam allow dispers interv mix remov test portion exce minut prepar primari dilut raw pasteur milk transfer steril pipett ml sampl raw milk pasteur milk ml diluent avoid contact pipett diluent temperatur diluent approxim milk sampl mix primari dilut carefulli mixer second primari dilut obtain prepar decim dilut raw pasteur milk transfer steril pipett ml primari dilut ml diluent instruct dilut obtain repeat oper obtain decim dilut number micro organ expect obtain inocul petri dish raw milk transfer steril pipett ml sampl decim dilut dish dilut examin prepar dish dilut chosen pasteur milk transfer steril pipett ml sampl decim dilut dish dilut examin prepar dish dilut chosen uht treat steril milk examin incub dai oc direct annex chapter vii point transfer steril pipett ml milk sampl dish prepar dish pour pour ml medium inocul dish mix immedi pour rotat petri dish suffici obtain evenli dispers coloni incub time end prepar milk sampl mix kind milk test portion dilut medium exce minut solidifi clean cool horizont surfac incub petri dish transfer dish incub incub dish invert stack high stack dish separ wall top incub incub oc hour count coloni count coloni petri dish coloni examin dish subdu light facilit count suitabl len talli counter avoid mistak particl precipit matter dish pinpoint coloni examin doubt object carefulli len higher magnif requir distinguish coloni foreign matter spread coloni consid singl coloni quarter dish overgrown spread coloni count coloni unaffect part dish calcul number entir dish quarter dish overgrown spread coloni discard dish calcul express result raw pasteur milk count dish coloni number micro organ ml raw pasteur milk formula sum coloni count equal volum sampl plate number dish count dilut number dish count dilut dilut factor count obtain count signific figur digit round round figur immedi left pasteur milk dilut coloni dilut coloni number ml number ml number ml count report number micro organ millilitr ml reciproc lowest dilut factor count exceed coloni count calcul estim count multipli reciproc dilut factor report result estim number micro organ ml uht steril milk plate count coloni ml consid longer satisfi requir direct eec precis result internation accept collabor trial enumer micro organ plate count test oc scope field applic procedur specifi refer procedur enumer micro organ mean coloni count techniqu oc pasteur milk incub milk oc dai determin degre contamin pasteur milk psychotroph micro organ capabl multipli milk oc definit design micro organ mean organ form countabl coloni incub aerob condit principl pasteur milk incub oc dai defin volum milk sampl mix cultur medium petri dish incub oc hour coloni count number micro organ ml pasteur milk calcul apparatu glasswar usual laboratori equip apparatu hot air oven capabl oper oc autoclav capabl oper oc incub capabl maintain temperatur oc oc point ph meter temperatur compens acur ph unit water bath capabl oper oc len magnif len magnif talli counter mixer capabl mix ml milk sampl decim dilut ml diluent work principl eccentr rotat content test tube glasswar test tube suitabl closur suffici capac adequ head space mix ml primari dilut decim dilut flask capac ml tube ml capac hold cultur medium pipett plug cotton wool glass steril synthet materi unbroken tip nomin capac ml outlet diamet mm petri dish clear uncolour glass steril synthet materi bottom dish intern diamet mm intern depth mm minimum bottom irregular interfer count coloni steril glasswar glasswar steril procedur maintain oc hour hot air oven maintain oc minut autoclav autoclav care ensur adequ penetr steam equip steril contain tightli close flask loos lid glasswar steril autoclav dri vent steam pipett steril hot air oven cultur medium milk plate count agar composit yeast extract trypton glucos dextros skim milk powder agar depend gell properti agar water ml skim milk powder free inhibitori substanc verifi compar test skim milk powder free inhibitori substanc prepar suspend dissolv compon order yeast extract trypton glucos final skim milk powder water heat suspens assist procedur add agar heat boil stir continu agar complet dissolv steam minut filter filter paper check ph ph meter adjust ph steril oc solut mol sodium hydroxid hydrochlor acid distribut steril storag cultur medium dispens medium quantiti ml flask ml tube stopper flask tube steril autoclav oc minut check ph medium medium immedi store dark temperatur oc longer month prepar commerci dehydr cultur medium cultur medium prepar commerci dehydr medium follow manufactur instruct add skim milk powder dissolv compon adjust ph oc dispens steril store medium diluent pepton salin solut composit pepton sodium chlorid nacl water ml prepar dissolv compon water heat check ph ph meter adjust ph steril oc solut mol sodium hydroxid hydrochlor acid distribut steril storag diluent dispens diluent test tube quantiti steril tube ml diluent stopper tube steril autoclav oc minut check ph diluent diluent immedi store dark temperatur oc longer month prepar commerci dehydr diluent diluent prepar commerci dehydr tablet powder follow manufactur instruct adjust ph dispens steril store diluent procedur melt medium begin microbiolog examin melt requir amount medium quickli temper medium oc waterbath prepar milk sampl incub unopen packag pasteur milk imposs repres sampl ml hour oc incub incub mix milk sampl micro organ distribut evenli rapidli invert milk sampl contain time foam avoid foam allow dispers interv mix remov test portion exce minut prepar primari dilut transfer steril pipett ml sampl ml diluent avoid contact pipett diluent temperatur diluent approxim milk sampl mix primari dilut carefulli mixer second primari dilut obtain prepar decim dilut transfer steril pipett ml primari dilut ml diluent instruct dilut obtain repeat oper obtain decim dilut number micro organ expect obtain inocul petri dish transfer steril pipett ml sampl decim dilut dish dilut examin prepar dish dilut chosen pour pour ml medium inocul dish mix immedi pour rotat petri dish suffici obtain evenli dispers coloni incub time end prepar milk sampl mix dilut medium exce minut solidifi clean cool horizont surfac incub petri dish transfer dish incub incub dish invert stack high stack dish separ wall top incub incub oc hour count coloni count coloni petri dish coloni examin dish subdu light facilit count suitabl len talli counter avoid mistak particl precipit matter dish pinpoint coloni examin doubt object carefulli len higher magnif requir distinguish coloni foreign matter spread coloni consid singl coloni quarter dish overgrown spread coloni count coloni unaffect part dish calcul number entir dish quarter dish overgrown spread coloni discard dish calcul express result count dish coloni number micro organ ml pasteur milk formula sum coloni count equal volum sampl plate number dish count dilut number dish count dilut dilut factor count obtain count signific figur digit round round figur immedi left dilut coloni dilut coloni number ml number ml number ml count report number micro organ millilitr ml reciproc lowest dilut factor count exceed coloni count calcul estim count multipli reciproc dilut factor report result estim number micro organ ml precis result internation accept collabor trial vi enumer coliform coloni count oc scope field applic procedur specifi refer procedur enumer coliform pasteur milk mean coloni count techniqu oc definit design coliform mean bacteria oc form characterist coloni characterist coloni ferment lactos product ga condit principl defin volum milk sampl mix cultur medium petri dish incub oc hour characterist coloni count ident characterist coloni confirm test abil ferment lactos number coliform ml pasteur milk calcul apparatu glasswar usual laboratori equip apparatu hot air oven capabl oper oc autoclav capabl oper oc incub capabl maintain temperatur oc point ph meter temperatur compens accur ph unit water bath capabl oper oc wire needl made platinum iridium nickel chromium glasswar test tube suitabl closur capac ml hold confirmatori medium durham tube dimens test tube flask capac ml hold solid select medium pipett plug cotton wool glass steril synthet materi unbroken tip nomin capac ml outlet diamet mm petri dish clear uncolour glass steril synthet materi bottom dish intern diamet mm intern depth mm minimum bottom irregular interfer count coloni steril glasswar glasswar steril procedur maintain oc hour hot air oven maintain oc minut autoclav autoclav care ensur adequ penetr steam equip steril contain tightli close flask bottl loos lid glasswar steril autoclav dri vent steam pipett steril hot air oven cultur media violet red bile lactos agar vrbl agar solid select medium composit pepton yeast extract lactos sodium chlorid nacl bile salt neutral red crystal violet agar depend gell properti agar water ml prepar suspend dissolv compon water leav stand minut mix vigor check ph ph meter adjust ph boil oc solut mol sodium hydroxid hydrochlor acid bring rapidli boil swirl time time immedi dispens quantiti ml steril flask temper medium water bath oc steril medium check time medium hour prepar brilliant green lactos bile broth confirmatori medium composit pepton lactos dehydr ox bile brilliant green water ml prepar dissolv compon water boil check ph ph meter adjust ph steril oc solut mol sodium hydroxid hydrochlor acid dispens medium quantiti ml test tube durham tube stopper tube steril autoclav oc minut durham tube air bubbl steril check ph medium medium immedi store dark temperatur oc longer month prepar commerci dehydr cultur media cultur media prepar commerci dehydr media follow manufactur instruct adjust ph dispens boil steril store media procedur medium medium vrbl agar prepar milk sampl mix milk sampl micro organ distribut evenli rapidli invert milk sampl contain time foam avoid foam allow dispers interv mix remov test portion exce minut inocul petri dish ml milk sampl inocul transfer steril pipett ml milk sampl dish pour pour vrbl agar inocul dish quantiti ml mix immedi pour rotat petri dish suffici obtain evenli dispers coloni incub time end prepar milk sampl mix test portion medium exce minut prepar steril purpos control uninocul dish ml vrbl agar inocul dish solidifi clean cool horizont surfac medium set complet solidif pour ml vrbl agar surfac inocul medium solidfi incub petri dish transfer dish incub incub dish invert stack high stack dish separ wall top incub incub oc hour count coloni count coloni petri dish coloni count dark red colour coloni diamet mm surround precipit characterist coliform coloni characterist aspect differ colour size format precipit typic coloni carri confirmatori test confirmatori test indic carri confirmatori test suitabl number characterist coloni inocul tube brilliant green lactos bile broth wire needl incub tube oc hour coloni produc ga durham tube confirm coliform calcul express result count dish coloni confirmatori test appli calcul number coloni coliform percentag confirm coliform coloni number coliform ml pasteur milk formula total number coloni coliform connect found examin milk sampl ml number millilitr sampl examin ml count signific figur coloni digit round round figur immedi left count exceed coloni report result estim number coliform ml precis result internation accept collabor trial vii enumer somat cell procedur specifi procedur refer procedur enumer somat cell microscop method fluoro opto electron method microscop method scope field applic procedur specifi refer procedur enumer somat cell raw milk procedur specifi procedur enumer number cell milk sampl calibr check accuraci fluoro opto electron procedur definit procedur somat cell cell leucocyt epitheli cell nuclei stain distinctli methylen blue principl ml milk spread cm slide film dri stain count carri microscop number somat cell count defin area multipli work factor obtain number cell ml reagent analyt grade chemic dye solut composit methylen blue ethanol ml trichloroethan tetrachloroethan ml glacial acet acid ml warn tetrachloroethan poison prepar applic carri fume cupboard prepar mix ethanol trichloroethan tetrachloroethan bottl heat water bath oc add methylen blue mix carefulli cool refriger oc hour add glacial acet acid filter filter pore size micron store dye solut air tight bottl particl sediment form filter apparatu glasswar microscop magnif microsyring ml accuraci slide area mm mm mark film standard slide templat mm mm film level hot plate oc dry slide fan hairdryer dry film water bath capabl oper oc heat milk sampl stage micromet slide rule mm divis procedur milk sampl milk sampl test hour sampl storag temperatur sampl exce oc freez avoid prepar sampl laboratori heat sampl water bath oc mix carefulli cool temperatur microsyring calibr oc pre treatment slide clean slide ethanol dry dust free paper flame cool store box avoid dust prepar film remov ml milk sampl prepar microsyring carefulli clean syring contact milk place syring slide draw outlin shape mm mm fill area evenli dry film level hot plate complet dry film milk sampl prepar examin stain film dip dye solut minut dry complet fan requir dip film tap water surplu dye wash dry store protect dust calibr microscop field depend magnif chosen determin stage micromet slide diamet microscop field count calcul count cell microscop count cell cell nuclei count recogniz count half nucleu visibl microscop field count strip field middl film avoid count strip field select exclus peripher area film care prepar film reliabl result check month count part film count perform count microscop field spread system part film equal repres minimum number cell count microscop count somat cell standard automat mechan count procedur coeffici variat count ident sampl higher electron instrument coeffici variat milk sampl cell ml exce number somat cell count sampl characterist poisson distribut order meet repeat poisson distribut presuppos varianc standard deviat coeffici variat cv cv cv cv cv cv wm indic number particl cell count cv calcul work factor ml milk work factor calcul count strip film length strip count mm breadth strip correspond diamet microscop field determin stage micromet slide work factor work factor diamet microscop field mm determin stage micromet slide number strip count complet count microscop field middl film grid work factor work factor diamet microscop field mm determin stage micromet slide number field count calcul cell content number somat cell count multipli work factor obtain cell ml milk precis coeffici variat exce result internation accept collabor trial fluoro opto electron method scope field applic procedur specifi refer procedur proper calibr se count somat cell raw milk chemic preserv definit procedur somat cell particl minimum intens fluoresc due stain dna nucleu somat cell principl part sampl ml mix buffer solut fluoresc solut part mixtur transfer form thin film rotat disc serv object plane microscop cell produc electr puls amplifi record number somat cell print thousand ml reagent analyt grade chemic state water distil deioniz equival puriti buffer solut composit potassium hydrogen phtalat potassium hydroxid polyethyleneglycol mono tetramethylbutyl phenyl ether triton volum ml ph make water ml prepar individu compon mix airtight storag exce dai fluoresc solut stock solut composit ethidium bromid make water ml prepar ethidium bromid dissolv water storag light proof airtight bottl exce month fluoresc solut work solut ml stock solut mix buffer solut give ml work solut longer dai clean solut composit buffer solut ml polyethyleneglycol mono tetramethylbutyl phenyl ether triton volum ml ammonia volum ml make water ml prepar individu compon mix storag exce dai apparatu glasswar count instrument work fluoresc optic principl note prior instrument calibr relat volum particl count threshold level count made determin calibr apparatu accord manufactur instruct sampl cell content determin microscop procedur water bath circul capabl oper oc test tube seal approxim ml milk sampl sampl store low temperatur test tube sampl chemic preserv count hour milk count low storag temperatur exce oc preserv chemic preserv hour preserv carri sampl chemic preserv sampl achiev addit preserv orthobor acid final concentr orthobor acid sampl exce ml preserv sampl store hour oc potassium dichrom final concentr potassium dichrom exce ml preserv sampl store hour oc sodium azid sampl preserv sodium azid final concentr ml provid sampl cool oc immedi sampl count hour sampl bronopol sampl preserv bronopol final concentr ml provid sampl cool oc immedi sampl count hour sampl sampl preserv orthobor acid preserv hour potassium dichrom note local condit discharg effluent observ sampl preserv potassium dichrom procedur pretreat sampl milk examin store milk hour approxim oc count sampl dai milk advis pretreat result low count sampl pretreat hour potassium dichrom prepar pretreat sampl untreat sampl dai heat water bath approxim oc sampl store room temperatur count cell count count carri count instrument minut end heat immedi count sampl mix order obtain homogen distribut somat cell dilut prepar sampl place automat instrument precis figur repeat reproduc intern collabor trial futur precis data state data nation level estim cell count level ml standard deviat repeat sr cell ml equival coeffici variat standard deviat reproduc sr cell ml equival coeffici variat control accuraci control accuraci perform sampl cell content determin microscop cell count nation refer laboratori viii detect antibiot sulphonamid scope field applic procedur specifi refer procedur detect antibiot sulphonamid raw milk heat treat milk refer procedur includ qualit method procedur initi procedur milk sampl antibiot includ sulphonamid select procedur number similar procedur principl bacillu stearothermophilu var calidolacti attc test organ procedur chosen repres test procedur confirm identif penicillin procedur confirm result qualit procedur identifi penicillin determin concentr penicillin qualit procedur scope field applic procedur specifi qualit detect antibiot sulphonamid raw heat treat milk excess limit laid tabl detect concentr antibiot sulphonamid test sensit neg posit benzylpenicillin ampicillin cloxacillin nafcillin tetracyclin oxytetracyclin chlortetracyclin chloramphenicol dihydrostreptomycin neomycin kanamycin bacitracin erythromycin rifamycin diaphenylsulfon sulphamethazin sulphadimidin benzylpenicillin bacitracin express iu ml antibiot mg ml definit milk antibiot sulfonamid colour medium chang principl milk sampl ad nutrient agar gel ph indic spore bacillu stearothermophilu var calidolacti atcc good sensit sensit inhibit penicillin incub result normal growth acid product organ ph indic colour chang purpl yellow presenc milk substanc inhibitori growth organ ph indic colour remain purpl apparatu glasswar usual laboratori equip apparatu incub capabl maintain temperatur oc water bath capabl oper oc rack tube ampoul pipett singl servic dispos tip suitabl sampl dispens ml forcep tweezer hot air oven capabl oper oc autoclav capabl oper oc ph meter glasswar sampl bottl suitabl closur note rubber stopper deposit inhibitori substanc neck bottl petri dish clear uncolour glass steril synthet materi flat bottom uniform thick minimum intern diamet mm bottl capac ml pipett plug cotton wool glass steril synthet materi nomin capac ml ml glass spatula tube ampoul intern diamet mm cap stopper steril glasswar glasswar steril procedur maintain oc hour hot air oven maintain oc minut autoclav autoclav care ensur adequ penetr steam equip steril contain tightli close flask bottl loos lid glasswar steril autoclav dri vent steam pipett steril hot air oven media solut test organ ingredi media suitabl bacteriolog purpos water glass distil deminer equal puriti substanc inhibitori test organ media nutrient agar composit yeast extract pepton meat extract sodium chlorid agar water ml prepar dissolv compon water bring boil swirl time time adjust ph steril oc dispens ml quantiti test tube make agar slant ml quantiti bottl steril oc minut agar medium composit sodium chlorid agar water ml trimethoprim tetroxoprim solut ml prepar dissolv compon trimethoprim tetroxoprim water bring boil swirl time time add trimethoprim tetroxoprim steril oc minut adjust ph steril oc trimethoprim tetroxoprim solut composit trimethoprim mg tetroxoprim mg ethanol ml ml water ml prepar dissolv trimethoprim tetroxoprim ethanol ml dilut water nutrient composit yeast extract mg glucos mg solubl starch mg bromocresol purpl water ml prepar dissolv nutrient indic water heat steril filtrat nutrient commerci tablet standard penicillin solut prepar solut penicillin mg ml iu ml dissolv crystallin sodium potassium benzyl penicillin steril distil water suitabl stopper steril bottl prepar work solut penicillin make ml penicillin solut ml steril distil water work solut mg iu ml prepar ml standard penicillin solut mg ml iu ml ad ml inhibitor free milk ml work penicillin solut mix offici journal european commun penicillin solut mention prepar dai test carri inhibitor free milk prepar control inhibitor free milk reconstitut skim milk powder previous test found free inhibitori substanc steril distil water altern suffici quantiti fresh bulk milk test found free inhibitori substanc dispens bottl heat hour oc store refriger oc maximum period week test organ bacillu stearothermophilu var calidolactu strain atcc test organ strain ident prepar stock cultur mainten test cultur test cultur slant nutrient agar agar slant surfac streak inocul loop test cultur incub aerob hour oc incub tube seal steril rubber stopper stock cultur obtain month refriger oc test cultur spore suspens ml nutrient agar transfer asept steril petri dish cool room temperatur steril pipett transfer ml steril distil water tube stock cultur wash spore agar slant steril loop spore suspens oc hour steril pipett transfer ml spore suspens cultur plate spread inoculum surfac bent glass rod incub oc hour stock cultur cultur hour subcultur procedur carri interv hour subcultur transfer steril pipett ml distil water cultur plate remov spore surfac suspens glass rod transfer spore suspens bottl ml steril distil water close bottl shake cultur subcultur immedi store refriger oc spore suspens viabl coloni count million ml plate count agar medium incub oc hour spore suspens uniformli turbid flock sediment discard suspens prepar stock cultur prepar test tube ampoul melt agar medium cool oc add part fresh spore suspens part agar medium tube bottl mix transfer ml inocul media calcul give layer mm thick steril tube ampoul close stopper cap melt tip test tube ampoul cool upright posit medium solidifi stand hour test tube ampoul dai month provid cool immedi prepar oc procedur sampl test prefer hour sampl maintain sampl oc meantim test sampl hour maintain deep freez oc minim inactiv penicillin identifi tube ampoul legibl indel remov cap stopper place requir number sampl control examin suitabl rack add microlit nutrient mention tube ampoul mix milk sampl transfer syring ml label tube ampoul clean dispos tip sampl transfer repeat oper duplic standard penicillin solut mg ml iu ml penicillin milk sampl repeat oper duplic control inhibitor free milk milk sampl close tube ampoul place rack tube ampoul water bath oc hour remov rack tube ampoul water bath observ color test medium interpret result purpl color test medium milk sampl control tube ampoul presenc antibiot sulphonamid posit level tabl page sampl color tube ampoul standard penicillin solut remain purpl prove test medium suffici sensit purpl color part test medium irregular color milk sampl tube ampoul inhibitori substanc level tabl page sampl yellow color test medium milk sampl control tube ampoul absenc substanc inhibitori test organ purpl color test tube ampoul includ neg control tube ampoul viabl spore sampl retest freshli prepar test materi confirm result confirm sampl reaction accord method storag milk sampl confirm deep frozen prevent degrad antibiot procedur confirm penicillin determin concentr scope field applic procedur specifi confirmatori test penicillin antibiot penicillin procedur determin penicillin concentr milk sampl posit dubiou reaction sensit antibiot procedur definit milk sampl antibiot includ sulphonamid sampl procedur clear zone mm inhibit disk sampl antibiot includ sulphonamid penicillinas betalactamas ad clear zone clear zone smaller diamet penicillinas inhibitori substanc penicillin penicillin antibiot includ sulphonamid zone inactiv penicillinas inhibitori substanc milk sampl penicillin residu direct eec annex chapter vi semi synthet penicillin sodium cloxacillin partli inactiv penicillinas complet resist identifi penicillin principl disk absorb paper impregn milk examin surfac agar medium inocul bacillu stearothermophilu var calidolacti incub result normal growth organ agar cloudi presenc milk substanc inhibitori growth organ clear zone disk size clear zone depend thing concentr type inhibitori substanc milk apparatu glasswar equip apparatu water bath capabl oper oc glasswar paper disk inhibitor free diamet mm capabl take approxim mg milk prefer store dessic media standard solut penicillinas solut reagent test organ ingredi media suitabl bacteriolog purpos water glass distil deminer equal puriti substanc inhibitori test organ media nutrient agar test medium detect inhibitori substanc composit yeast extract trypton glucos trimethoprim tetroxoprim solut ml agar depend gell qualiti water ml prepar solid compon complet dissolv water heat stir trimethoprim tetroxoprim solut ad ad trimethoprim tetroxoprim solut ph adjust steril oc medium steril minut oc standard penicillin solut milk quantif inhibitori substanc make standard penicillin solut inhibitor free milk concentr mg ml iu ml mg ml iu ml mg ml iu ml mg ml iu ml penicillinas solut dissolv suffici penicillinas betalactamas steril distil water give concentr ml solut prefer divid small portion store oc week note uniform intern standard penicillinas purpos procedur assum unit penicillinas suffici inactiv mg iu penicillin suppli penicillinas unknown strength check assumpt valid chang concentr penicillinas solut correspondingli penicillinas solut commerci disk prepar penicillinas control procedur found amount penicillinas test organ test cultur spore suspens prepar test plate melt test medium detect inhibitori substanc coolto oc add bottl part fresh spore suspens part test medium detect inhibitori substanc densiti coloni inocul test medium mix transfer steril petri dish previous heat oc inocul test medium give layer mm thick petri dish mm intern diamet ml test medium order obtain thick mm transfer petri dish cold horizont surfac previous check spirit level remov lid agar medium solidifi medium solidifi lid replac dish invert minim condens surfac agar medium test plate prepar prefer dai week provid seal polyethylen bag oc immedi prepar order identifi sampl mark bottom test plate procedur prepar sampl sampl give posit dubiou result method retest identifi quantifi penicillin initi milk sampl heat oc minut avoid influenc thermolabil specif inhibitor mix approxim ml heat test milk transfer suitabl steril wide mouth bottl ml penicillinas solut ad milk mix detect inhibitor dip paper disk milk sampl clean dry pair forcep remov excess milk touch disk side sampl bottl place disc flat surfac test plate press gentli forcep disk prepar milk sampl mm mm edg order check sensit disk dip standard penicillin solut randomli milk sampl disk rate number milk sampl disk standard disk test disk agar medium random fashion identifi invert plate incub oc hour incub plate examin front suitabl light sourc clear zone inhibit paper disk clear zone measur zone disk penicillin standard solut mm clear zone disk milk sampl size greater substanc inhibit test organ identif quantif inhibitori substanc procedur perform duplic heat milk sampl sampl treat penicillinas ad penicillinas ml milk sampl prepar penicillinas disk dip sampl test plate procedur perform duplic standard penicillin solut mention averag diamet clear zone inhibit milk sampl penicillinas control standard penicillin solut determin interpret result clear zone disk penicillinas control clear zone disk milk sampl equal greater zone disk standard penicillin solut inhibitori substanc milk sampl correspond sodium potassium benzyl penicillin concentr mg ml averag diamet clear zone disk penicillinas equal averag diamet clear zone disk milk sampl milk inhibitori substanc imposs inactiv penicillinas concentr procedur averag diamet clear zone disk penicillinas smaller averag diamet clear zone disk milk sampl heat accord milk sampl penicillin antibiot includ sulphonamid penicillin semi synthet penicillin identifi penicillinas concentr procedur synthet penicillin sodium cloxacillin inactiv penicillinas condit test classifi inhibitor penicillin note inhibitori substanc penicillin requir identifi suitabl procedur determin penicillin content determin penicillin content perform draw standard curv calcul zone size obtain standard penicillin solut milk draw standard curv linear correl log penicillin concentr diamet inhibitori zone standard curv drawn semilogarithm paper penicillin concentr logarithm ordin inhibitori zone abscissa inhibitori zone calcul averag duplic test diamet inhibitori zone plot standard penicillin concentr standard curv drawn calcul penicillin concentr milk sampl calcul zone diamet equat standard curv accur assai radiu inhibit zone time bigger time radiu disk express result result express content penicillin equal mg ml indic determin concentr content inhibitor penicillin repeat reproduc figur meaning standard includ comparison ix detect pathogen micro organ scope field applic accord direct eec requir annex chapter vii procedur instruct check pasteur milk pathogen micro organ definit examin made bacteria speci frequent involv food born diseas pasteur treatment safeguard presenc thermoresist pathogen milk standard laid annex chapter vii direct coloni count oc oc coliform phosphatas met specif test pathogen suspicion milk connect food poison outbreak procedur procedur frequenc examin laid nation author make issu health certif heat treat milk intra commun trade detect pathogen micro organ appli criteria procedur internation accept result report pathogen micro organ investig result express number ml milk presenc absenc volum pasteur milk requir procedur report describ procedur patent legisl cultur media anti folat substanc observ