National Dairy Development Board For Efficient Dairy Plant Operation my Jechnews ay -Augast 1999 Mo. 21 RAPID MICRO-BIOLOGICAL TESTS ‘This bulletin includes technical information, latest development on products, ystems, techniques etc. reported in journals, companies’ leaflets an books and based on studies and experience. The technical information in differen isues ison diferent area of plant operation. eis hoped that the information contained herein, if employed inthe dairy plans, will help in making it's operations more efficent. Your contributions and suggestions will make the bulletin more useful and are welcomed. ‘The theme of information inthis issue is Rapid Microbiological Tests. {Tr may be noted that the information given hee is by no means com plete. Duthis iste 1. Betroduction Tachnologiee Rapid tects for millcandmilk products awe 206‘LINTRODUCTION ‘2. TECHNOLOGIES ‘ecaniy oevie a compte nthe Imeem wordt api aus chage ‘Wi he wore wrens fee food for consumer, the Codex ‘Alimentarie Commision hi ‘ecommesded employing the Hazaré ‘Avalysis Creal Contol Po (GIACCP) Syste i food root Indeed, ic his become esental to Implement HACCP Systm a dairy frocesng oe Teche ine 11,12, Tan 20, Proverive adel comeetve scons Meese tccnsre rode) and tlio to facilitate cost contol by ‘minimialng downgraded product, sework andlor acrapped product and ‘nimizing dowe-tme and cleaning etron proces equpme ‘Speed Is now the vital factor for checking seepule microbial levels, ‘monitoring hypiee and relesting Finished products trom storage ‘Tradiions| mirobiologicl teste Ihomener tke 2103 6ay6 Rap est mets bye, eeore,becote very imporanio fe emloyedin dry pant. laoduton of pid miero-ilogial test meds ry pln in Idi is ‘able frp micoboingeal ets of reduc and oequipment seics fr hei yin canon. apd mirobilogial mets can be ‘powped nour rsd cone |. and tomate waona ethos etc counting eto: ‘ethos ao the mestrement ethods deeting bo-chemicl mpoest of miro Modi aod Automated Traitonal Methodst ‘These metods offer Simplified, more wer Iienly Convenience devices or automatic ‘stems for simpler reprinting rovers ndeouning However ere [Sho benefit the time taken 0 obin ‘ere. Direct Cell Counting Methods: These methods include the Direct pierce iter Techigue (DEFT) ‘casts vetoes wing ese ‘inp and Mow eye “They offer he pel of very rapid alps sed ive erate i 20°30 ns Thay howovereuic cole, atu, skilled operion and subuancal sample preperation, DEFT ‘sige forthe sly ak Tithe been ecefely ena a the ecg is limited wo her Nonaenous fee noeet rete ‘nel, These meted ao have the Palen deel end enumerate thespecific poops of micro-orgmisos ‘tough ew ea routine ase. “Methods based om the Measurement “Metabolic Activity: This can be (ide tbo pt. (1) simple meio or (2) optical methods ‘The simple matte ae based on te ol detrinn geicper an tet ae 2:8 hour. Alina Hinge and ches, ‘hey nck sensi and ety Te Sophisticated methods. include alert ices whic lyon the ably of sotvely metabolizing ‘meo-opsins to prods dezetble ‘ional such a elec conductance, eat riballed carbon oxide ot rest changes Impedance bas fond ‘veranda aplctions and ges ‘erent ore. Biosensors bt ‘bepromiscf snl portable device tat Ine not ee developed wih siickent sens and ead, Methods Detecting Biochemical Componeats of Micro-organlems: ‘These etd inde ATP (eneine ic) Soluminessence several other tecbooogies sich a ELISA (Eeuyme Linked “Immunosorbant Assays) and LAL (Limalus Amoebocye Lyste) tes ‘deetng pects ergaisns aa mater ‘ftoun TheLAL at deste entin of gram nepave tates and ELISA ett paogenc bce Salmoneli ‘They rede vel eeton te by 23 daya by sp the fea ‘eteton and iemifcsion steps. The ‘eeu recat advances have prodoed Sp ue ny inl ocd formas for aed components and no epi ovement, a recent ine, ATP bihminecence technigu has gained wide popelrty fed sceptics All iving el coxtaia [ATP and ATP bioluminescence test Getects ATP using firefly enzyme Tura. The fell wt 6 ted ‘he measrernen of ight greeted by Iseiern/acferaretescon Sa the fresence of ATP to quantify the ‘Sotogia resides preset nthe test sample sey apd nd este and ‘esis ad fod rede, ving teal fn mines, A wide range of ‘pplication iclde porablebyene (Eyicee fp monitor te fn and performance ofeletingprocedies, Tnetremente to tat raw milk and ‘tomate struments 9 ey product (gay ater proce ‘Teemost widely acre an sed rapid Imethods in mlrobiology abroad are Snpedence, ATP Mslmieecece sed BISA, Most of the inetrmetsaasbe for pid microbiologist ‘ilk products ae iter rect cll Counting methods (DEFT, flowcytometry) or methods detec boshemial components of micokes (ATP ELISA) Istrarments te vale {or measuring ttl bacterial cots (1BCx) snr specie microbe come Information ie provided er ely 08 Incruments wich mei TBC. BACTOSCAN: Bactscan FC, fom FOSS, ie aizect cell couating ‘esrument Dated on flow eytomety Thais verysubleopcasysem eich ver av improved sigaoise ratio Increasing the sensivty. The Insane nearer sore wife time in excess of 10,000 hrs. The Sncwmet i ed fr milk kes « 445 ml milk sample, which passer hough eit be throughly mie tn homogenied. The incubation feagnt s aed ndthe mixed SC fort mots, Dring ie, ‘he raaton pe several tines hog ‘Bnemesh ites eas diinepsion ofsomatccel, pte an nt lbs fd thorough sing of bse ik the fluorescent stan (Ethidiom bromide). The mistare then passes ‘hugh fw cll where ser Bee exces igi pt ule er oraisn, Is convened int eleonie pulse and ‘counted. Te result i expresied as {advil base celle pr ml “The intament is availabe in tree sie, with houghpts of $0, 10 an 150 samples pr hau -Asoter version it Bactotcan #000, ‘ich bar foe houghpat capacity ‘han Bretoscan FC. This diet cell coun inetraent tthe Drops (Ging Wace icine. is sed to ‘mea ttl vale comms (TVS) of ‘retrain milk (CELSIS-LUMAC: The Luma Raw Mile Microbial Kit with Blocounter "M1800 M2500 i tse on the ATP technique. Whea an mikaivs atthe (hiya eampe canbe ete in wound 5 minutes, The system measures the ‘ctl coot inte of Reeve Light ‘site (RLU), and then eaibrted in ‘terms of Colony Forming Uais (CFU) 4000 ie also. based on ATP ‘otaminescence ecniqoe The sample ‘eadatobe wine fora peridot oly “boar, Then heres ae obined fromm te asrment ihn minute. Tee inouent marr he ete count se RLU and is calbated to diploy rer a pas or al Another instrament based on ATP Bioluminescence technique. from Cals Lat rte Parteerand Mik ‘Screen Kit wih Bioeounter M180, 1M 2800 oF Autobincounter M400 (Fig), Ate 24h peincabtion period, pastearied milk ample it (etd ine intranet in mines mae lo offer api ing oe Salmonella, Patl-Stk One Step Salmonella Test.Figure I: The Aatebocnnter M4000 CHEM BLOW: The Chemfiow ‘Astosyiem 3 of Pa-based company (Chemaner slo dct cl couing Insrument taden daw cfomeny. The Inseumeet has pk sensei, and ‘hooghpatof sales per hour Even Stout peenrihment, ican delet levee of 100 eran pe i st than 30 mnie Catton sganeme ambe detested wii Irs snd yeasts Sn eas thn 24s In Oeinseument he bed client a tuoescen signal enim the lcer team 0 each als cued irc by serie opal detec. ‘Chenfow 3h ten we formic tes fori, erste foe UT ra doer et BIOCOM: Antero cel counting Inerrment avi is Asteria 1 from Fench Company Bice. Aste ‘Sattereyomety baredecnigo bt Uses an advanced system. Sample insruen sale samples of ior total bacterial counts (TBC) snd ‘pecfecel numbers Aste hae he pct cy ot 400 ane per tour Biocon’ another sii, hough ess vanced, instumeat for mess ‘abate oun of ak Caen ‘apd micobiclogy sym This he futonated version of the DEFT tec BIOIRACE: The Biotrace Dairy Kit snd Microbial Lamineseence Sytem is'bared on ATP bioluminescence Technique fo. detect microbial “uy preduca. The neuen at Nghly sensitive state-of the luminometer and ele reese it the iegral compote. UAT products detest ae preieabred for 48-72 132830 Deg C. The nseunen get ‘el thn 20 misnteasipe YES orNO.‘OTHERINSTRUMENTS: Sane ster Jnstunens sng ATP iolsmineceace ‘ToFin2) capac ofa widerngetiat {oe ay proses. Ten sree for pasearmaon of mil ona ele in milk, insecucides, aflatoxin, Picci qaliy and yee. Pir 2: The Portable Pllunisometer ‘The LUMET weigh only 09 Bas 35 Sterna batery a igh en, Sinpltoread LCD dpa tar only ‘inimtes perform a 3tep ATP tt Rens ean be expressed os RLU, ‘pene on, pl pass ite Bioprobe, with appropiate kis, om HaghesWhitoc,can meno bceral oun (a tem of RLU) rile, ‘ee eam an ter daly pose 4. RAPID HYGIENE ‘MONITORING SYSTEMS ‘There ace a umber f mpi hygienic ‘rontoring iret sale which fe portable, simple and exe wo we Mose thm are ATP bilminercence based. Some of them a briefly senuoae here. BIOTRACE: For ssresing the lemliness of equipment suc, the fnerument of Botace, UK ae the ‘ot popu ee Te inst re Based ‘on ATP bioleminescence tecnigue 1s UniLite Fp) war among the ist direct, portable hygiene sab test inunentstbeintoduced Ipovies ‘estrella of eco Fay {Wes obutandigh weigh tives ‘elnblerehs (psf) wien an be displceé on its LCD (Liguid {Cys Display) seen raha copy ona micropriner Powered ectarge: ‘Bie fullin Date oransAC power, Iti sible owe bots rodcton ‘Unit Lite Kelis an advanced pore Fes 3 “Taesysem al compas sa loging capaiies Rechargeable ary enabler Melty of pean Alta we suomaicalyrcrded Seconds with tbe dt ine, user and Pn et Ree shown a palFig. 3: Biorace Unt end ‘net XCEL cata/a wih stn securiy that ‘heer esto ackaowedge sal Als ‘ean perorm wed snl. ‘To make seabbing simple ad rapid, Bjorace Ha iavotuced Clea (Big, which baseline Sige ‘Sots will expen inset The [remind sah removed ron ts bide the test autace under tert fabbed and the swab returned by ‘eprteingacliponthe Clan Trace ant hing i ly down Ti acts {he tet for measuemest The Cea ‘race eine no he Unite Lite Xeel sample chamber and rest ‘eu, Tet ee un 30 eco ‘Seabeanbescbvaad uot howler swabans ‘CELSISLUMAG: Celsis Lamae's ashe Rinse Water Test Kit can te eed to check onthe opt whether the CIP of processing equipment as een effect cent Tt ives ares inves thf teresa robe de ‘Sinaegonciesing cree ton faa be taken immediatly. A simlar ‘nsrument is Hen Monitoring Ki. ‘These tee ed combo With Fiure 4: Blaroce Clean Trace Lama's Bigomater MISO or M2500 Relative Light Unite (RLU) and ‘sted in Pasi. Te armen te ATP aro Aoter hae gine monting Steen! os Cell SpotCheck ‘Thi ingle shot system ute peated Colour change tecology to sect he ‘pesnce of AP. The cups imple ‘hourchinge ative the ated yer, SpotCheck employe a sovel fated enzyme packaged into asl alaloed, single sho posable pene ape device The deve reno ‘The reals ae ip wa siple Cele change tn ensperent t ark (geen atocurs wii Io 1 ines,depending eee entation ‘Sst Sure si other ATP bed portable inirsent 1 np les an ‘oo gar 3 simple seein sing ‘Aa AIP.e rah tiene wh ‘Swabbiagsluion an then th tet we is evabbe. The swab ppd into 8 ‘ube coining enzyme eeegent and ated fr abet 10 tc Asap placed onthe take and the tube Inserted ino the chaer fhe Sates insramentsavalable fr equipment jpienemonterng Laminaae-san [ATP bused iasrament fam Ameren ‘company Charm is liphweigt, porable system which ser the Pocketswaby a singloass swab with ulin eagen ht ese asoor, ‘empertre fr wpto one month and ves el in 30 secede: Resa a (ven aps and sured a RLUs. ‘The Hy-Lite ATP monitor is from Germat Company Merck, ‘Thi semprises x sampling pe, agai ae Sentinel ener devi: Resa ae read as pusicavion/fl, and the Instant hat memory for 1000, rent Azaheisrument is Blprobe for ‘Haphes Wack Company. Ile aah weight (2 kg) portable ATP based Insouneat. ei ene at Ieel of ‘tra slow a6 10 el pr gem. can be denied sng this ses 100 seen, Sapmese Konic’ simple, epi and atts ystemi¢ Sab N Check, ‘hs ted ren reader ot entre of the hygiene eats of work Srnes. The testa cloinet a tet wich gives a colour change ro ‘wees wo darks parle in he peseace {€ peptide bonds. The rele re ‘hind in shoo 10 mince ‘Aodhersmils isrument ie Check Proton Diveey Lever, Lighining Fg) fom US Company exe ins ATP based hand-held laste with neat ro,‘REFERENCES ‘lorie Clean-Trae, lt MLM O86 ese No.2 Bios Li Bgen ux Biounc. Unb-Lite Xed, leat MLM No 108 Isve No. Biotace Lid, Boge’, UK, (Celis (1897 Sytem Sure Portable Hygiene Monier, net PIC 9012/ E CasLamae, Cambie Lasse HACCP and the Lumae Solution, elt C7002, Ces, Lams, [Endgaa, The Nededands mse. Product Information, Leet CRS/Sch/2120/E Celis Lame, anda, The Nera (dra Chem (1958). Raps eins or ptearntion smell hyn, “Mik inusry Tt, Fb Russel, P1999). Inseumenttion for he dir nbortory, Mik Industry Tn, Mach Rosel (1997 Monitoring hyiee, Mk industry Tt, Tn Russ, P. (1958. New Ingedies for fod profucts, Mik Industry It, Be 10, Ruse, 195). Rapist mee evewed, MK Indasty Int, Ap find his buen 1 ven TD tative Baring ony enterainng 1 ink i forma of his tin eds no need changes | woud ke information nan beset sve on___ ‘Please send your ters to: ‘Regional Director (SR) ‘Naor! Day Develpoen Bod 'PBNo9S06, VI Block, 30 Fee: Root KORAMANGALA BANGALORE 56 095