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Effects of Inulin and Oligofructose on the Rheological Characteristics and Probiotic Culture Survival in Low-Fat Probiotic Ice Cream
A.S. AKALIN AND D. ERIS IR
ABSTRACT: The effects of supplementation of oligofructose or inulin on the rheological characteristics and survival of Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 in low-fat ice cream stored at 18 C for 90 d were studied. Addition of oligofructose or inulin to ice cream mix significantly increased apparent viscosity and overrun and developed the melting properties in ice cream during storage (P < 0.05). However, the highest increase in firmness, the lowest change in melting properties, and the longest 1st dripping time were obtained in probiotic ice cream containing inulin (P < 0.05). Some textural properties have also improved especially by the end of storage. Freezing process caused a significant decrease in the viability of Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 (P < 0.05). Oligofructose significantly improved the viability of L. acidophilus La-5 and B. animalis Bb-12 in ice cream mix (P < 0.05). Although the viable numbers for both bacteria decreased throughout the storage, the minimum level of 106 CFU/g was maintained for B. animalis Bb-12 in only ice cream with oligofructose during storage. Keywords: Bifidobacterium animalis Bb-12, inulin, Lactobacillus acidophilus La-5, low-fat ice cream, oligofructose

M: Food Microbiology & Safety

airy products with incorporated probiotic bacteria are gaining popularity and the probiotics comprise approximately 65% of the world functional food market (Agrawal 2005). The species of bacteria most commonly used in dairy products for probiotic effect are Lactobacillus and Bifidobacterium (Saxelin and others 2005). Standards requiring a minimum of 106 to 107 CFU/g of Lactobacillus acidophilus and/or bifidobacteria in fermented dairy products have been introduced by several food organizations worldwide (Shah 2000). Therefore, it is important to ensure a high survival rate of these bacteria during the product shelf life to maintain consumer confidence in probiotic products (Saxelin and others 1999). Ice cream seems suitable for delivering probiotics in human diet because of its pleasant taste and attractive texture. However, in order to ensure that the product provides an adequate content of microorganisms, cells must survive in freezing and frozen storage. Freezing and thawing cause various degrees of damage to cells, including microorganism death, inhibition of its development, reduction, or interruption of metabolic activity (Davies and Obafemi 1985). Recent studies have focused on the survival of probiotic bacteria in ice cream produced by different techniques such as culturing ice cream mix (Hekmat and McMahon 1992; Davidson and others 2000; Akn 2005; Favaro-Trindade and others 2006), nonfermented ice cream mix (Alamprese and others 2002; Haynes and Playne 2002), or adding fermented milk to regular ice cream mix (Christiansen and others 1996; Hagen and Narvhus 1999). Nondi-

gestible food ingredients or prebiotics that selectively stimulate growth and/or activity of probiotic bacteria have been used to increase the viability of probiotic bacteria in dairy products. On the other hand, by decreasing the fat content in frozen dairy product formulations, quality characteristics on body and texture are affected (Ohmes and others 1998). In this respect, inulin and oligofructose, the best-known prebiotics and also fat replacers, possess several functional and nutritional properties that may be used to formulate innovative healthy foods for todays consumer. Inulin is a term applied to a heterogeneous blend of fructose polymers found widely distributed in nature as plant storage carbohydrates. It has a degree of polymerization (DP) of 2 to 60. Oligofructose is a subgroup of inulin, consisting of polymers within a DP 10. Both inulin and oligofructose are widely used in functional foods throughout the world (Sangeetha and others 2005). Their structure is similar to corn sweeteners, principal carbohydrates used in ice cream technology. Classed as fat replacers, inulin and oligofructose influence the bulk and mouthfeel of the products. Also they are resistant to hydrolysis in both the stomach and small intestine, and are classified as dietary fiber ingredients (Spiegel and others 1994; Niness 1999). The main uses of inulin and oligofructose are as texturizing agents, particularly in low-fat foods such as ice cream (Devereux and others 2003). Some studies have been reported on the functionality of inulin as a fat replacer in reduced fat ice cream (SchallerPovolny and Smith 2001), in yog-ice cream (El-Nagar and others 2002), and in fat-free starch-based dairy dessert (Tarrega and Costell 2006). However, no research has been reported on both functional and prebiotic effects of inulin and oligofructose as a food MS 20070590 Submitted 7/27/2007, Accepted 2/7/2008. Authors are with Ege ingredient in low-fat probiotic ice cream. Thus, our objective was to Univ., Faculty of Agriculture, Dept. of Dairy Technology, 35100, Bornova, compare the effects of inulin or oligofructose supplementation esIzmir, Turkey. Direct inquiries to author Akaln (E-mail: sakalin21@ pecially on the survival of probiotic starter culture and also the rheyahoo.com). ological characteristics of low-fat probiotic ice cream. In addition,

Introduction

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C 2008 Institute of Food Technologists doi: 10.1111/j.1750-3841.2008.00728.x

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Probiotic culture survival in ice cream . . .


Overrun% = (weight of ice cream mix) (weight of ice cream) we aimed to compare the rheological characteristics of regular low 100 (weight of ice cream)1 (Marshall and others 2003). fat ice cream and probiotic ice cream. Apparent viscosities of the mixes were evaluated at 4 C after Materials and Methods 24 h aging using a Brookfield RV Viscometer fitted with spindle no:5 at 20 rpm (Brookfield Engineering Laboratories, Stoughton, Mass., Ingredients and formulation for ice cream U.S.A.). Results were multiplied by RV viscometer factor (2000/N , In the production of ice cream, cows milk was supplied from N = 20 rpm) and given as Pa.s. Firmness of ice cream was deEge Univ., Agricultural Faculty, Menemen Research Farm (Izmir, termined by a Surberlin PNR 6 Penetrometer (Sommer Runge KG, Turkey), pasteurized cream containing 35% milk fat, and non- Berlin, Germany). Penetrations of a conical spindle weighing 91.6 fat milk powder was supplied from Pnar Dairy Industry (Izmir, g ( 0.1 mm) to ice cream at 18 C were measured after 5 s. BeTurkey), and freeze-dried DVS starter cultures of Lactobacillus fore the measurements were taken, penetrations of the probe were acidophilus La-5 and Bifidobacterium animalis subs. lactis Bb- conducted 4 cm from the side of each cup and were repeated twice. 12 were obtained from Chr. Hansen Lab. (Hoersholm, Denmark). Firmness was measured as the depth (in mm) of penetration of conOther ingredients for low-fat ice cream mix included sucrose and ical spindle into the ice cream and then a firmness index (g/mm) corn syrup (G-40) (Cargill, Istanbul, Turkey), stabilizeremulsifier was calculated by dividing the conical spindle weight (91.6 g) to the mixture of Cremodan SE 30 (Danisco AS, Copenhagen, Denmark), depth of penetration (mm). inulin (Fibruline XL, molecular weight: 3300, degree of polymerMelting behavior, expressed as 1st dripping time and melting ization > 20), and oligofructose (Fibrulose F97, molecular weight: properties, was evaluated on ice cream samples stored at 18 C. 1000, degree of polymerization < 20) (Cosucra AS, Fontenoy, Bel- Melting properties were determined by carefully cutting the plastic gium). cups from the ice cream samples (preweighed as 25 g), placing the ice cream onto 1-mm stainless steel screen over a cup, and weighManufacture of ice cream ing the amount of ice cream drained into the cup over a 90-min All ice creams were manufactured in the pilot plant of Dairy period at 20 0.5 C. The time for the 1st drop of melted ice cream Technology Dept., Faculty of Agriculture, Univ. of Ege. Mix formu- was also determined (Christiansen and others 1996). lation was 4% (w/w) milk fat, 12% (w/w) milk solids nonfat, 13% (w/w) sucrose, 0.65% (w/w) stabilizer/emulsifier, 4% (w/w) 42 Dex- Enumeration of probiotic bacteria trose Equivalent corn syrup for regular ice cream (R) and probiotic The count of viable probiotic bacteria was determined after agice cream (P), 4% (w/w) oligofructose for probiotic ice cream with ing of the mix for 24 h at 4 C, and then during the storage days of oligofructose (PO), and 4% (w/w) inulin for probiotic ice cream the samples. One gram of probiotic ice cream sample was diluted with inulin (PI). with 9 mL of sterile 0.1% (w/v) peptone water (Oxoid, Basingstoke, Raw milk and cream were weighed into stainless steel milk Hampshire, U.K.) and mixed uniformly with a vortex mixer. Subsecans. All dry ingredients were mixed into the cold liquid ingre- quent serial dilutions were made and viable cell numbers enumerdients and complete incorporation was ensured. The mixes were ated using the pour plate technique. The counts of L. acidophilus pasteurized at 68 C for 30 min. L. acidophilus La-5 and B. an- La-5 were enumerated on MRS agar (Merck, KGaA 64271, Darmimalis Bb-12 cultures were added to the mixes (0.3%) except for stadt, Germany) incubated aerobically at 37 C for 72 h. (Christhe regular sample (R), after cooling to 40 C, to achieve ap- tiansen and others 1996). B. animalis Bb-12 was enumerated acproximately 108 CFU/g, mixed well, and fermented for approx- cording to the method of Lankaputhra and others (1996) using imately 4 h at 40 C until the desired pH of 5.5 was reached. MRS-NNLP (nalidixic acid, 15 mg/L; neomycin sulphate, 100 mg/L; Hekmat and McMahon (1992) reported that probiotic ice cream lithium chloride, 3 mg/L and paramomycin sulphate, 200 mg/L) was preferred at pH 5.5 regarding overall acceptance by judges. agar. Filter sterilized NNLP was added to the autoclaved MRS base The fermented mixes were then cooled in an ice bath to 5 C. just before pouring (Laroia and Martin 1991). The inoculated plates All mixes were aged at 4 C for 24 h to ensure complete hydra- were incubated anaerobically at 37 C for 72 h using an oxygen tion of all ingredients. Mixes were frozen in random order using free gas mixture of anaerobic jars (Merck). Plates containing 25 to a batch ice cream freezer (4 L capacity, U gur, Nazilli, Turkey) for 250 colonies were enumerated and recorded as logarithm of colony 35 min. The ice cream was packaged into 150-mL plastic cups and forming units (CFU)/g of sample. 50-mL plastic cups (as 25 g for melting behavior), and then placed in a hardening room at 18 C. The experiment was conducted in Statistical analysis triplicate. Each experiment was independently replicated 3 times and all analysis and enumerations were done in duplicate. Analysis of variCompositional analyses ance for each set of data was conducted as a factorial arrangement Total solids in the ice cream was determined by drying the sam- of treatments in a completely randomized block design to deterples for 3.5 h at 100 C and fat contents were analyzed by means of mine whether significant differences existed. For the storage exthe Gerber method (AOAC 1990). The pH values of ice cream sam- periment, the set of data was conducted as a split plot in a ranples were measured with a pH-meter combined with a glass elec- domized complete block design. Each replication was a block; milk trode (Beckman Zeromatic SS-3, Beckman Instruments Inc., Fuller- treatment was the main unit treatment, and days of storage were ton, Calif., U.S.A.). The titratable acidity in ice cream was deter- the subunit treatment. The model equation was Yi jk = + i + mined with N /10 natrium hydroxide in the presence of phenolph- j(i) + k + ( )ik + i jk where , i , j (i ) , k , ( )ik , and i jk reprethalein and expressed as percent lactic acid. sent overall mean effect, effect of milk treatment i, random effect of block j receiving milk treatment i, effect of storage time k, Rheological analyses milk treatment by storage time interaction, and experimental error, Overrun was measured with a comparison of the weight of ice respectively. Data were analyzed using the general linear model procedure of cream mixture before and after freezing. The formula for overrun is the SPSS Win 9.0 program, and Duncans multiple range test was as follows:
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Probiotic culture survival in ice cream . . .


Table 1 --- Results (mean SD, n = 3) of compositional and physical analyses on aged mix and ice cream. Mix Mix or ice cream R P PO PI pH 6.90 0.00 5.52 0.05a 5.52 0.05a 5.47 0.05a
b

Ice cream Total solids (%) 33.38 0.04 33.42 0.02a 33.47 0.11a 33.49 0.05a
a

Apparent viscosity (Pa.s) 1.76 0.0 2.68 0.03b 3.35 0.01c 3.91 0.04d
a

Fat (%) 4.0 0.0 4.0 0.1a 4.1 0.1a 4.1 0.1a
a

Lactic acid (%) 0.14 0.01 0.51 0.01b 0.51 0.01b 0.52 0.02b
a

pH 6.90 0.01 5.45 0.06a 5.45 0.17a 5.35 0.17a


b

Overrun (%) 23.6 4.0a 27.6 1.9ab 31.7 1.3b 50.6 2.5c

R = regular mix or ice cream; P = probiotic mix or ice cream; PO = probiotic mix or ice cream with oligofructose; PI = probiotic mix or ice cream with inulin. a,b,c Means with different letters in the same column are different (P < 0.05).

used to compare means when the effect was significant (P < 0.05). In addition, statistical significance was given in terms of P values, with differences at the 95% confidence interval (P < 0.05) being considered statistically significant (SPSS 1997).

Results and Discussion


ompositional analyses of ice cream samples performed in the 1st day of storage revealed that the targeted total solids and fat levels were achieved (Table 1). As expected, pH and lactic acid contents of regular and probiotic ice cream samples were significantly different (P < 0.05) while similar lactic acid contents and pH values were determined in all probiotic ice creams (P > 0.05). Regular ice cream sample had a mean pH value of 6.90 0.01 and lactic acid percentage of 0.14 0.01. There were significant differences in viscosities among all mixes, including probiotic ice cream mixes, and viscosity increased by addition of oligofructose or inulin to mix (P < 0.05) (Table 1). High apparent viscosity in the probiotic ice cream mix containing oligofructose or inulin can be explained by the interactions of the dietary fiber and liquid components of the probiotic ice cream mix. Ice cream mixes containing carbohydrate-based fat replacers exhibit a viscous behavior because of the capability for imbibing water, which would increase the viscosity of the system (Schmidt and others 1993). The highest mean apparent viscosity of 3905 MPa.s (P < 0.05) was obtained in the probiotic mixes containing inulin (Table 1). Similar to our findings, significantly higher apparent viscosity was obtained by replacing 100% of the 42 DE corn syrup with inulin in a reduced fat ice cream mix (Schaller-Povolny and Smith 2001). The authors reported that higher apparent viscosity resulted from the higher molecular weight of inulin and that a potential interaction between the inulin and milk proteins could also be present in the system. Higher molecular weight of inulin may be related to higher apparent viscosity of the ice cream mix with inulin in our study. Inulin, being highly hygroscopic, would bind water and form a gellike network that, in addition to other components (like corn syrup or emulsifierstabilizer mixture), would modify the rheology of the mix. Similar results in relation to the effect of inulin on viscosity were also reported by El-Nagar and others (2002) and Akn (2005) for yog-ice cream and probiotic-fermented ice cream, respectively. The highest overrun value was also obtained in probiotic ice cream mix containing inulin (P < 0.05), indicating its responsibility for the increased air incorporation (Table 1). The overrun value increased approximately 2 times when inulin was used in the manufacture, in contrast to the findings of Akn (2005) for probioticfermented ice creams. The addition of L. acidophilus La-5 and B. animalis Bb-12 and fermentation of the mix did not significantly affect the overrun values (Table 1). Alamprese and others (2002) also reported that Lactobacillus johnsonii La1 addition did not modify the overrun of ice cream. In the current study, a direct correlation has been determined between firmness and melting behavior. Our results indicated that all probiotic ice creams were found to be firmer than regular ice
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Figure 1 --- Firmness index of ice cream during storage. R = regular ice cream; P = probiotic ice cream; PO = probiotic ice cream with oligofructose; PI = probiotic ice cream with inulin. The error bars represent the standard deviation (n = 3). a,b,c Means with different letters in the same storage day are different (P < 0.05).

cream. Addition of oligofructose or inulin increased the firmness in probiotic ice cream. (P < 0.05) (Figure 1). However, ice cream supplemented with inulin was significantly firmer than other products throughout the storage except the 1st day (P < 0.05). Due to its longer chain length, inulin is less soluble than oligofructose and has the ability to form inulin microcrystals when sheared in water or milk. These crystals interact to form a creamy texture (Niness 1999). In addition, the ability of inulin to bind water molecules and form a particle gel network can improve the firmness of the product (Franck 2002). Although it seems that firmness was improved in all products by extension of storage, significant increases were not found (P > 0.05) except for the 60th day for the P sample, the 60th and 90th days for the PI sample, and the 90th day for the R and PO samples. A slower change in melting properties was observed in probiotic ice creams when compared to control sample during storage (P < 0.05) (Figure 2). Melting properties were also improved by using oligofructose and inulin (P < 0.05). However, the most remarkable improvement in melting behavior was obtained in the product containing inulin (P < 0.05). The change in melting properties decreased in all samples as storage time increased, and the least change was obtained at the 60th and 90th days for the PI sample and the 90th day for the other products (P < 0.05). The 1st dripping time was also longer in probiotic ice creams supplemented with oligofructose and inulin in comparison to the control sample (Figure 3). However, inulin increased the 1st dripping time more than oligofructose, which was found to be statistically significant for all storage days (P < 0.05). Additionally, the times prolonged in all samples as storage time increased while the

Probiotic culture survival in ice cream . . .


longest time was reported at the 60th and the 90th days for the R, P , and PO samples and at the 90th day for the PI sample (P < 0.05). Typically, ice crystal size increases by about 30% to 40% during hardening of ice cream. In the storage, ice recrystallization occurs. The small crystals melt at the same time that large crystals grow. The changes in ice crystals due to the thermodynamic ripening process are enhanced by temperature fluctuations. Small crystals, with a slightly lower melting point, are more sensitive to temperature fluctuations than larger crystals (Marshall and others 2003). Inulin or oligofructose can control ice recrystallization like a stabilizer agent. Therefore, the 1st dripping time of all samples can be improved by these interactions as storage time increased. Addition of inulin led to the lowest change in melting properties and longest 1st dripping time as well as the most increase in firmness (P < 0.05), due probably to the high molecular weight and hygroscopic properties of inulin. The gelling properties of inulin improve the consistency of mix and retard the melting of the product. These observations are consistent with those of El-Nagar and others (2002) who demonstrated that inulin supplementation reduced the melting rate and increased firmness in yog-ice cream. Akn (2005) also reported that addition of inulin retarded the melting time of probiotic-fermented ice cream. This study has verified that the highest values for the apparent viscosity, overrun, and firmness and the most remarkable improvement in the meltdown characteristics were obtained in the mix or ice cream containing probiotics and inulin (P < 0.05). Ice creams containing a high amount of air (high overrun) tend to melt slowly. Air cells act as an insulator (Marshall and others 2003). The viable counts of probiotic bacteria were 7.74 0.51, 8.44 0.16, and 8.24 0.04 log CFU/g for L. acidophilus La-5 and 7.58 0.62, 8.49 0.14, and 8.120.28 log CFU/g for B. animalis Bb-12 in the ice cream mixes P , PO, and PI, respectively. When compared to the control sample, the viable counts for both L. acidophilus La-5 and B. animalis Bb-12 significantly increased in the probiotic ice cream mix by addition of oligofructose (P < 0.05) due to the possible prebiotic effects of oligofructose in the ice cream mix. Fructooligosaccharides (FOS), especially oligofructose, are preferred by bifidobacteria as a source of carbon and energy. Growth rates of bifidobacteria cultivated on either oligofructose or inulin were evaluated and better growth was obtained on oligofructose than inulin (Wang and Gibson 1993; Gibson and Wang 1994). In addition, in vitro fermentation of inulin revealed that molecules with a shorter chain length are fermented quicker than molecules with a longer chain length (Roberfroid and others 1998). Therefore, higher survival of these probiotics in ice cream mix containing oligofructose can be sourced from shorter chain length or lower polymerization degree of oligofructose than inulin. The changes in the viable counts of L. acidophilus La-5 and B. animalis Bb-12 in ice cream samples during storage are presented in Table 2. During freezing of the mix, the counts of both viable bacteria decreased by 1.5 to 2.0 log units, and their numbers in the frozen ice cream were found to be in the range of 5.96 to 6.60 log CFU/g for B. animalis Bb-12 and 5.98 to 6.21 log CFU/g for L. acidophilus La-5. The decline in bacterial counts, as a result of freezing, is most likely due to the freeze injury of cells leading eventually the death of cells. Furthermore, the incorporation of oxygen into the mix may have resulted in an additional decrease in viable cell counts as well as the mechanical stresses of the mixing and freezing process. The counts also significantly decreased (0.3 to 0.9 log CFU/g) throughout the storage (P < 0.05); however, freezing and mixing involved in converting the mix into ice cream had a greater effect on culture viability than storage in ice cream (P < 0.05). A similar finding was reported by Hagen and Narvhus (1999), Alamprese and others (2002), and Haynes and Playne (2002), but not by Hekmat and McMahon (1992). During freezing and storage of ice cream, more or less reduction in the survival of probiotic bacteria was also reported (Hekmat and McMahon 1992; Christiansen and others 1996; Hagen and Narvhus 1999) for different microorganisms, different production technologies and formulations, and pH. On the other hand, Davidson and others (2000) and Alamprese and others (2002) reported that starter culture bacteria in low-fat ice cream did not change significantly during storage. In our study, B. animalis Bb-12 survived better than L. acidophilus La-5 in ice cream over 90 d (Table 2). However, the viable counts of B. animalis Bb-12 were higher than the recommended minimum limit of 106 CFU/g only in ice cream containing oligofructose during storage. In addition, according to the general mean value of storage, the ice cream products supplemented with oligofructose contained higher viable counts of both probiotic bacteria during the storage, possibly depending on the higher viable
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Figure 2 --- Melting properties of ice cream during storage. R = regular ice cream; P = probiotic ice cream; PO = probiotic ice cream with oligofructose; PI = probiotic ice cream with inulin. The error bars represent the standard deviation (n = 3). a,b,c,d Means with different letters in the same storage day are different (P < 0.05).

Figure 3 --- First dripping times of ice cream during storage. R = regular ice cream; P = probiotic ice cream; PO = probiotic ice cream with oligofructose; PI = probiotic ice cream with inulin. The error bars represent the standard deviation (n = 3). a,b,c Means with different letters in the same storage day are different (P < 0.05).

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Probiotic culture survival in ice cream . . .


Table 2 --- Viable counts of L. acidophilus La-5 and B. animalis Bb 12 (mean SD, n = 3) in ice cream during storage (log CFU/g). Ice cream 1st day 5.98 0.25aB 6.21 0.02aB 6.00 0.09aB 6.27 0.19bB 6.60 0.20cB 5.96 0.13aB 30th day 5.53 0.18aAB 5.77 0.11bA 5.47 0.14aA 5.97 0.07bA 6.40 0.17cAB 5.36 0.35aA 60th day 5.02 0.48aA 5.79 0.15bA 5.24 0.10aA 5.93 0.26bA 6.45 0.28cAB 5.51 0.19aAB 90th day 5.13 0.28aA 5.70 0.10bA 5.12 0.46aA 5.94 0.20abA 6.25 0.11bA 5.47 0.55aAB Mean of storage 5.41 0.48a 5.87 0.23b 5.46 0.41a 6.03 0.23b 6.43 0.22c 5.57 0.39a

L. acidophilus La-5 P PO PI B. animalis Bb-12 P PO PI

P = probiotic ice cream; PO = probiotic ice cream with oligofructose; PI = probiotic ice cream with inulin. a,b,c Means with different letters in the same column are different (P < 0.05). AC Means in the same row with different superscripts are signicantly different (P < 0.05).

counts in mixes with oligofructose and more conducive structure of oligofructose to cell viability during storage (P < 0.05). The lowest viable counts of B. animalis Bb-12 obtained in the products with inulin in the first and other days of storage (P < 0.05) can be caused by the higher overrun rate of these samples (Table 2). In general, being strictly anaerobic, Bifidobacterium spp. are more sensitive to oxygen than L. acidophilus (Talwalkar and Kallasapathy 2003).

he best improvement in textural characteristics in terms of firmness, melting properties, and 1st dripping time was obtained in probiotic ice cream with inulin during storage (P < 0.05). Viable starter culture counts reduced (0.3 to 0.9 log CFU/g) during the storage, but at the initial freezing and churning stage of converting mix into ice cream a greater decrease (1.5 to 2.0 log unit) in the counts L. acidophilus La-5 and B. animalis Bb-12 (P < 0.05) was observed. Survival of L. acidophilus La-5 and B. animalis Bb-12 in ice cream was significantly enhanced with oligofructose (P < 0.05) and the recommended minimum limit of 106 CFU/g was maintained for B. animalis Bb-12 in only probiotic ice cream with oligofructose during storage.

Conclusions

Acknowledgments
The authors thank the Ege Univ., Faculty of Agriculture, and Research Fund Council for financial support to this study.

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M: Food Microbiology & Safety

M188

JOURNAL OF FOOD SCIENCEVol. 73, Nr. 4, 2008