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Thermal Inactivation of Escherichia Coli in Camel Milk

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Thermal Inactivation of Escherichia Coli in Camel Milk 1708 Journalof Food Protection, Vol. 66, No. 9, 2003, Pages 1708– 1711 Copyright q,InternationalAssociation for FoodProtection ResearchNote Thermal Inactivationof Escherichia coli inCamel Milk SHLOMO SELA, * RIKY PINTO, UZI MERIN, AND BARUCH ROSEN Departmentof Food Science, AgriculturalResearch Organization,The Volcani Center, P.O. Box 6, Bet Dagan50250, Israel MS02-431:Received 26November 2002/ Accepted 28February 2003 Downloaded from http://meridian.allenpress.com/jfp/article-pdf/66/9/1708/1676688/0362-028x-66_9_1708.pdf by guest on 28 September 2021 ABSTRACT Camelssubsist and produce milk in desert pastures not utilized by other domesticated herbivores. Developing the camel milkindustr ycanimprove the economy of desert inhabitants. T ocomplywith sanitary ordinances, camel milk is pasteurized byprocedures speci ed for bovine milk. It iswidelyaccepted that milk composition might affect bacterial thermal death time (TDT). Cameland bovine milks markedly differ in their chemical composition, yet data regarding TDT valuesof bacteriain camelmilk is missing. As a rststep toward developing speci c heattreatments appropriate for camel milk, TDT curvesof Escherichiacoli inarti cially contaminated camel and cow milks have been compared. Heating the milks to temperatures rangingfrom 58 to 65 8Cyieldssimilar thermal death curves and derived D- and z-values.These ndingssuggest that, in this temperaturerange, E. coli mightbehave similarly in bovine and camel milk. Additional TDT studiesof various pathogenic speciesin camel milk are required before establishing pasteurization conditions of camel milk. Camelssubsist and produce milk in desert pastures not femalesmaintained in the Central Arava V alleyin southernIsrael utilizedby other domesticated herbivores. Developing the betweenJanuary and August 2002. Milking was done twice a day, camelmilk industry can help in improvingthe economy of morningand evening. A sampleof the pooled, mixed milk was desertinhabitants. Recently, in the south of Israel, camel cooledimmediately postmilking and kept at 4 8C.Every4weeks, growersare commercializing camel milk production. The samplesof themorning milk were transferred to the laboratory in marketingof camel milk imposes on it legal regulations; arefrigeratedcontainer immersed in crushed ice within 4 hafter amongthese are compliance with sanitary indices, such as milking.The microbial quality of raw camel milk brought to the totalbacteria and coliform counts. At present,those indices laboratorywas assayed on standard plate count agar (PCA; La- areenforced by ordinancescontrolling the quality and safe- boratoriosConda, Spain) for total aerobic bacteria count, and vi- olet-redbile agar (VRBA; LaboratoriosConda, Spain) for coli- tyof bovine milk in terms of microbialindices. T ocomply form counts (9). PCA plateswere incubated at 32 8Cfor48 h,and withthe ordinances, camel milk is pasteurized by proce- VRBA plateswere incubated at 32 8C for 24 h. duresspeci ed for bovinemilk. Lately, a few lotsof pas- Thecomposition of the camel milk from the supplying herd teurizedcamel milk were rejectedby the authorities be- wasanalyzed by standard methods (9) atthe Dair yScienceLab- causeof noncompliance with microbial standards. It was oratory(DSL), TheV olcaniCenter ,Israel.The composition of necessaryto know whether this was becauseof improper bovinemilk was analyzed by the DSL andby the central testing dairymanagement, faulty pasteurization, postpasteurization laboratoryof the Israeli Cattle Breeders Association, Caesarea, contamination,or anothercause. A lackof dataon the ther- Israel. maltreatment of camel milk was soonapparent. No valid Theraw milk was pasteurized by heating 50-ml aliquots in reason,scienti c orlegal-formal,could be foundwhy camel glassbottles at 65 8Cfor30 min. The bottles were then cooled to milkshould be pasteurizedin conditions applying to bovine 48Candkept refrigerated. All thermal death studies were per- milk.It is well established that camel milk differs inits formedon the pasteurized camel milk within a week.Pasteurized compositionfrom thatof bovine milk (3, 13) and that the bovinemilk was used as acontrol.The bovine milk was collected compositionof the suspending medium affects the fate of fromnumerous producers, standardized, pasteurized, and pro- heat-treatedbacteria (16). cessedby a largecommercial dairy. The milk was used within Acquiringknowledge on the time-temperature param- theexpiration date given by the manufacturer . etersapplicable to camel milk pasteurization is, therefore, anessential step in the commercialization of camel milk. Bacterialstrain. Theorganism used in all the TDT experi- ments was E. coli TG1 (AmershamBiosciences, UK) towhich Thefollowing study presents data on thethermal death time pMalCplasmid, carr yingampicillin resistance gene was intro- (TDT) of Escherichiacoli incamel milk. duced.Bacteria were grown in 4 mlof tryptic soy broth (TSB) MATERIALS AND METHODS supplementedwith 100 mg/mlampicillin for 20 hat37 8C, washed oncewith double-distilled water and resuspended in 10 mMphos- Milk. Camel (Camelusdromedarius )milkwas obtained from theInternational Camel Center herd, which included 20 milking phatebuffer (pH 7.2). Cold camel or bovine (3.6 ml) was inoc- ulated with E. coli (0.4ml) to yield a nalconcentration of ;108 *Authorfor correspondence. T el: 1972-3968-3750;Fax: 1972-3960- CFU/ml.Aliquots (100 ml) of E. coli-containingmilk in 0.2-ml 4428;E-mail: [email protected]. thin-walledThermo-tubes (ABgene, Surrey, UK) wereused in all J.FoodProt., Vol. 66, No. 9 THERMALINACTIVATION OF E. COLI INCAMELMILK 1709 TABLE 1. Bacterialcounts in raw camel milk raw camelmilk (65 8C,30 min), total counts were reduced 2 3 Milk Totalbacteria Coliformcount to 10 to 10 CFU/ml,while coliform counts were reduced sample (CFU/ml)a (CFU/ml) to ,1CFU/ml(data not shown). Universallyspeci ed conditionsfor milkpasteurization 5 4 1 1.41 3 10 6.80 3 10 were originallydesigned to eliminate Mycobacteriumtu- 2 2.44 3 104 8.00 3 103 berculosis andare also useful to eliminateother pathogenic 3 6.20 3 105 1.17 3 105 4 1.20 3 108 4.00 3 107 bacteria,such as Listeriamonocytogenes, Yersinia entero- 5 2.50 3 106 1.40 3 103 colitica, and Mycobacteriumparatuberculosis, as well as 6 5.50 3 106 9.00 3 102 pathogenicstrains of E.coli(6, 15). However,thepractical 7 8.00 3 104 5.00 3 103 dailyindices that assure milk supply safety rely on total 8 5.30 3 108 9.00 3 107 bacteriaand coliform counts (9). Theemergence pathogens, 9 4.70 3 106 8.50 3 105 such as E. coli O157:H7in milk supplies emphasizes the Downloaded from http://meridian.allenpress.com/jfp/article-pdf/66/9/1708/1676688/0362-028x-66_9_1708.pdf by guest on 28 September 2021 a U.S. Foodand Drug Administration grade ‘ ‘A’’pasteurizedmilk importanceof these tests. ordinance (12). ‘‘Individualproduce rmilknot to exceed Amajorexpected source of variation in our experi- 100,000/mlprior to commingling.’ ’ ‘‘Notto exceed 300,000/ ml mentswas thecamel milk. Changes in camel milk com- ascommingled prior to pasteurization.’ ’ positionduring the experiment could have modi ed theef- fectsof the heat applied to microorganisms suspended in it.The genetic history of the milking dames of the Inter- TDT experiments.The tubes were heat treated in a thermocycler nationalCamel Center herd, as it affects their milk, is un- equippedwith Peltier technology and a 96-wellsilver block (Biometra,Germany). The machine was programmed to hold sam- known.Effects of sire genetics on camel milk and the ples at 48Cfor5 min,heat them to the selected time-temperature breedinghistory of the individual dames are obscure. Lac- combination,and instantaneously cool them to 4 8C. tatingdames are affectedby internalfactors, such as sexual Thenumber of viable E. coli ineachsample was determined cyclesand individual metabolic differences and by external byplating 100 mlofserial dilutions of samples on both PCA factors, includingclimate,h uman-inducedstress ,and supplementedwith 100 mg/mlampicillin (PCA-amp) and VRBA. changesin feed and water (10). Feedsupplied to the herd Calculationof the D- and z-values. Tocalculatethe decimal was notwell controlled, and feed-milk relationships of the reductiontime ( D-value),the log 10 ofthe CFU representingor- camelare not clear .Indeed,the camel milk compositional ganismssurviving after heating was plotted against the measured studiesprove its expected heterogeneity (T able2). T oas- exposuretime. All calculations were done on that portion of the surethe veracity of theexperimental system and to separate TDT curvethat demonstrated a perceptiblecontinuous decline in artifactsfrom the‘ ‘true’’ results,it was desiredto constantly thecounted CFU. Linearregression was used to correlate the data challengethe experimental system by a controlthat had to points.The reciprocal of thegenerated slope of theregression line beconstant, homogenous, and similar to the tested system. wasused to state the D-value (11). Thelinear regression of the Itwas decidedthat a syntheticmedium would not do. The log10 of the D-valuesplotted against temperature was used to de- rive the z-valuefor degrees centigrade (1, 11). Statisticalanalysis Israelidairy cows have been genetically ‘ ‘standardized’’ for wasperformed by Microsoft Excel. over70 years. Cows are maintained in enclosed barns and arefed carefully composed feed designed to affect the com- RESULTS AND DISCUSSION positionand quantity of theirmilk. Consequently, the com- Microbialquality of raw camel milk. Bacterial positionof the Israeli bovine milk is uniform
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