LWT - Food Science and Technology 62 (2015) 682e688

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LWT - Food Science and Technology

journal homepage: www.elsevier.com/locate/lwt

Beneficial microorganisms viability and sensory acceptance of a

potentially synbiotic dairy-based tomato spread
Cristina Dini Staliano a, Rafael Chacon Ruiz Martinez b, S.M.I. Saad a, *
a
Department of Biochemical and Pharmaceutical Technology, School of Pharmaceutical Sciences, University of Sa~o Paulo, Av. Prof. Lineu Prestes,
580, B16, 05508-000 Sa~o Paulo, SP, Brazil
b ~o Paulo, Av. Prof. Lineu Prestes, 580, B14,
Department of Food and Experimental Nutrition, School of Pharmaceutical Sciences, University of Sa
05508-000 Sa~o Paulo, SP, Brazil

a r t i c l e i n f o a b s t r a c t

Article history: Nowadays, important features for the development of new food products include their convenience,
Received 31 July 2014 taste, and ability to promote beneficial effects to human health. This study aimed to develop a potentially
Received in revised form synbiotic dairy-based tomato spread and to evaluate the viability of the microorganisms employed, as
22 November 2014
well as the sensory acceptability of the product throughout its storage for 28 days. Three different trials
Accepted 12 December 2014
Available online 19 December 2014
were studied, all containing the prebiotic inulin and the starter culture Streptococcus thermophilus ST-M6:
T1 (control); T2 with the probiotic strains Lactobacillus acidophilus NCFM and Bifidobacterium animalis
subsp. lactis Bb-12, and T3 with Bb-12 and the bacteriocinogenic and potentially probiotic strain
Keywords:
Probiotic
Lactobacillus sakei subsp. sakei 2a. The product was stored at 4
C and analysed after 1, 7, 14, 21, and 28
Inulin days of storage. For all trials, the pH decreased throughout storage, the viability of all strains was high
Lactobacillus (7.9 log cfu/g), and sensory evaluation with consumers showed mean scores above 7 during the storage
Bifidobacterium period evaluated (up to 21 days), which demonstrates the product has potential as a functional food.
Dairy-based spread © 2014 Elsevier Ltd. All rights reserved.

1. Introduction
The recent trend in the development of new food products is
their convenience, taste, and human health promoting function-
ality. A functional synbiotic product combines probiotic and pre-
biotic ingredients, which may improve the gastrointestinal tract
(GIT) function. Probiotics are defined as live microorganisms, which
when administered in adequate amounts confer a health benefit on
the host (FAO/WHO, 2001). Prebiotics are selectively fermented
ingredients that promote specific changes in the composition and/
or activity of the GIT microbiota and confer benefits to the host
(Gibson, Probert, Van Loo, Rastall, & Roberfroid, 2004).
Lactobacillus acidophilus NCFM and Bifidobacterium animalis
subsp. lactis Bb-12 are commercial probiotic cultures used in a va-
riety of food products and dietetic supplements. Dairy products are
considered good vehicles for probiotics delivery and, among them
cheese is the most appropriate product, due to its buffering ability.
In general, cheeses show higher pH when compared to fermented
milks and, therefore, are able to protect the microorganisms against
* Corresponding author. Tel.: þ55 11 3091 2378; fax: þ55 11 3815 6386.
E-mail address: susaad@usp.br (S.M.I. Saad).
the harsh conditions found in the GIT, helping to maintain high cell
viability rates (Cruz, Buriti, Souza, Faria, & Saad, 2009). Tomato
(Lycopersicon esculentum) is an interesting natural ingredient to be
added to a dairy-based product. It contains different antioxidants,
including lycopene, b-carotene, and vitamin C. Lycopene has been
shown to be useful to prevent different types of cancer by reducing
the side effects of free radicals and unstable molecules, which could
damage the body's healthy cells (Lugasi, Ho  va
ri, Biro
, Brandt, &
Helyes, 2004).
Some probiotic and non-probiotic lactic acid bacteria (LAB)
strains are able to produce antimicrobial compounds, known as
bacteriocins (Nishie, Nagao, & Sonomoto, 2012). Bacteriocins are
ribosomally synthesized antimicrobial peptides and a promising
technological alternative for preventing the growth of spoilage and
pathogenic microorganisms in foods, a process called bio-
preservation (Snyder & Worobo, 2014). Lactobacillus sakei subsp.
sakei 2a is an aerotolerant LAB isolated from a Brazilian fresh
sausage (De Martinis & Franco, 1998) and there is only one study
available in the scientific literature about its application in a dairy
product (Villarreal et al., 2013). Lb. sakei subsp. sakei 2a is able to
produce bacteriocins (Carvalho et al., 2010) and its probiotic po-
tential was shown in gnotobiotic mice challenged with the food-
borne pathogen Listeria monocytogenes (Bambirra et al., 2007).

http://dx.doi.org/10.1016/j.lwt.2014.12.030
0023-6438/© 2014 Elsevier Ltd. All rights reserved.
 C.D. Staliano et al. / LWT - Food Science and Technology 62 (2015) 682e688 683

Therefore, this study aimed to develop a particularly innovative cheese; salt; olive oil; commercial sucrose; stabilisers guar gum,
synbiotic dairy-based tomato spread and to evaluate the viability of xantan gum, carrageenan gum, prebiotic fibre inulin, and the whey
the microorganisms employed, as well as the sensory acceptability obtained during cheese production. After the mixture step, a
of the product throughout its refrigerated storage for up to 28 days. smooth salmon cream was formed. Samples containing approxi-
mately 40 g of the food product were packaged in individual plastic
2. Material and methods pots for food products (68 mm diameter, 32 mm high, 55 mL total
volume, Tries Aditivos Pl ~o Paulo, Brazil). The plastic pots
asticos, Sa
2.1. Experimental design and microbial cultures were sealed with aluminium covers, and stored at 4 ± 1
C for up to
28 days. Table 2 shows the dairy-based spread overall formulation.
Experimental design was randomized and consisted of three
combinations (trials) (Table 1). Every trial was manufactured as 2.3. Determination of pH
three repetitions (triplicates) and samples from the food product
were obtained at 1, 7, 14, 21, and 28 days of storage for further The pH values of the dairy-based tomato spreads were deter-
analysis. mined for triplicate samples with a Thermo Orion Three Stars pH
The strains Streptococcus thermophilus ST-M6, B. animalis subsp. metre (Thermofisher Scientific, Waltham, MA, USA) equipped with
lactis Bb-12 (Christian Hansen, Hoersholm, Denmark), and Lb. aci- a penetration electrode model 2AO4 (Analyser, Sa ~o Paulo, Brazil)
dophilus NCFM (Danisco, Dange , France) used in the present study after 1, 7, 14, 21, and 28 days of storage at 4 ± 1
C and also during
consisted of freeze-dried commercial cultures powders for direct the steps of the quark cheese preparation.
vat inoculation (DVS) and were added at 0.01 g/100 mL, 0.03 g/
100 mL, and 0.05 g/100 mL, respectively. Lb. sakei subsp. sakei 2a
2.4. Microbiological analysis
was kept in De Man Rogosa Sharpe (MRS) broth (Oxoid, Basing-
stoke, UK) added of glycerol (20 mL/100 mL) and stored at e 80
C.
Microbial counts of Lb. acidophilus NCFM, B. animalis subsp. lactis
The microorganism was grown in MRS broth at 30
C for 24 h, re-
Bb-12, Lb. sakei subsp. sakei 2a, and St. thermophilus were con-
cultivated (inoculum of 4 mL/100 mL) at 30
C for approximately
ducted throughout the food product storage (after 1, 7, 14, 21, and
16 h, centrifuged (5000  g for 10 min) and finally washed twice
28 days) for all trials. For the microbiological analysis, portions of
(NaCl 0.9 g/100 mL) to obtain the pellet. This washed pellet was
25 g of the food product were transferred to aseptic bags (Nasco,
inoculated directly in the 18 L of milk employed for the cheese
Whirl-Pak®, USA), added of 225 mL of 0.1 g/100 mL peptone (Oxoid)
production.
water, blended in a Bag Mixer 400 device (Interscience, St. Nom,
France), and then submitted to further ten-fold serial dilutions
2.2. Dairy-based tomato spread production using the same diluent.
St. thermophilus ST-M6 was counted by pour-plating 1 mL of
Quark cheese was produced with 18 L of skimmed pasteurised each dilution in agar M17 (Oxoid) supplemented with 10 g/100 mL
milk (Salute, Descalvado, Brazil; high temperature short time of lactose (Oxoid), followed by aerobic incubation at 37
C for 48 h
[HTST] process), which was heated to 37e38
C and inoculated (Richter & Vedamuthu, 2001). Lb. acidophilus NCFM was enumer-
with the appropriate microbial cultures, according to the experi- ated by pour-plating 1 mL of each dilution in formulated MRS agar,
mental design shown in Table 1 and described above. Next, calcium modified by the replacement of glucose by maltose, followed by
chloride (0.25 g/L) was added in all trials. As soon as the pH reached aerobic incubation at 37
C for 48 h (International Dairy Federation,
values in the range of 6.3e6.5, commercial rennet Ha-la (100% 1995). B. animalis subsp. lactis Bb-12 populations were determined
chymosin identical to the bovine enzyme; Christian Hansen, by pour-plating 1 mL of each dilution in MRS agar (Oxoid)
Valinhos, Brazil; 5 mg/L) was added to the cheese-milk, which was
allowed to set again for approximately 1 h, until a curd was formed
and the pH reached values between 5.6 and 5.8. The gel formed was Table 2
cut into cubes (20  20  20 mm), placed in sterilised cotton Ingredients and formulation of the dairy-based spreads studied.
cheesecloth, and allowed to drain at 15
C for 15 h. After draining, Ingredients Amount (g/100 g)
the cheese-base was placed in sterilised beakers, covered with a
1. Quark cheese 50.00
PVC film and stored at 4 ± 1
C, until the final mixture took place. 2. Tomato pulp 14.00
For the final mixture, an industrial blender (Lucre, Catanduva, 3. Milk cream 13.00
Brazil, 15 L capacity) was employed for mixing the following in- 4. Dried tomato 11.00
5. Inulin 4.00
gredients: quark cheese; pasteurized tomato pulp; dried tomato;
6. Whey 2.60
commercial sterilized milk cream; dried basil; grated parmesan 7. Parmesan cheese 1.51
8. Olive oil 1.50
9. Sugar 1.00
Table 1 10. Salt 0.80
Description of the three different trials (T1, T2, and T3) of the dairy-based tomato 11. Guar gum 0.22
spread studied, according to the microbial cultures employed. 12. Basil 0.15
13. Xantan gum 0.11
Trials St. thermophilus B. animalis subsp. Lb. acidophilus Lb. sakei subsp.
14. Carrageenan gum 0.11
ST-M6a lactis Bb-12b NCFMc sakei 2ad
Total 100.00
T1* þ e e e
2. Pasteurized tomato pulp (5.8
Brix, Pomodoro, Goia ^nia, Brazil); 3.
T2 þ þ þ e
Commercial sterilized milk cream (25% lipids, Nestle , Araçatuba, Brazil);
T3 þ þ e þ
4. Dried tomato (Agro Industrial Itaguaçú, Itaguaçú da Bahia, Brazil); 5.
*
control; þ ¼ addition; - ¼ no addition. Prebiotic inulin (GR, Orafti, Oreye, Belgium); 7. Grated parmesan cheese
a
Streptococcus thermophilus ST-M6 (Christian Hansen, Hoersholm, Denmark). (Faixa Azul, Vigor, S~ ao Paulo, Brazil); 8. Olive oil (Gallo, Mairinque,
b
Bifidobacterium animalis subsp. lactis Bb-12 (Christian Hansen). Brazil); 9. Commercial sucrose (Uni~ ao, Limeira, Brazil); 10. Salt (Cisne,
c
Lactobacillus acidophilus NCFM (Danisco, Dange , France). Cabo Frio, Brazil); 11. Guar gum (Grindsted® 250, Danisco, Cotia, Brazil);
d
Lactobacillus sakei subsp. sakei 2a (Laboratory of Food Microbiology, School of 12. Dried basil (Fuchs, Itupeva, Brazil); 13. Xantan gum (Grindsted® 80,
Pharmaceutical Sciences/University of Sa ~o Paulo, Sa
~o Paulo, Brazil). Danisco), and 14. Carrageenan gum (Grindsted® CY500, Danisco).
684 C.D. Staliano et al. / LWT - Food Science and Technology 62 (2015) 682e688
supplemented with both sodium propionate (3 g/L) and lithium
chloride (2 g/L) (MRS-LP agar), followed by anaerobic incubation
(Anaerogen Anaerobic System, Oxoid) at 37
C for 72 h (Vinderola &
Reinheimer, 1999). Lb. sakei subsp. sakei 2a was counted by pour-
plating 1 mL of each dilution in MRS agar (Oxoid), followed by
aerobic incubation at 30
C for 48 h (De Martinis & Franco, 1998).
2.5. Sensory evaluation
According to the experimental design (Table 1), three trials (T1,
T2, and T3) were evaluated during storage. Sensory evaluation was
conducted with consumers (“untrained panellists”) after 7, 14, and
21 days of storage of dairy-based spreads at 4 ± 1
C. In total, 304
healthy adult consumers participated in the study. In order to
guarantee an adequate statistical analysis, the minimum number of
different consumers required for every section was estimated as 50.
For consumers' selection, the inclusion criteria were people aged
between 18 and 60 (belonging to both genders) and that had
tomato-based products consuming habits. The exclusion criteria
consisted of individuals with symptoms of allergy, food intolerance
or chronic disease, and also under medical treatment, sick (flu and
cold) or indisposed. The study was approved by the Ethics Research
Committee from the School of Pharmaceutical Sciences of the
University of S~ao Paulo.
For all nine sensory analysis sections conducted, product sam-
ples of approximately 20 g were served in plastic vials identified
with a three-digit random code number. The samples were stored
in a refrigerator (4 ± 1
C), and immediately taken from there to be
offered monadically to the consumers, who evaluated the product
through an acceptability test using a nine-point structured hedonic
scale [ranging from “like extremely” (9 points), “neither like nor
dislike” (5 points) to “dislike extremely” (1 point)] (Lawless &
Heymann, 1999). Most of the consumers participated in more
than one session and were instructed to report the sensory attri-
butes related to flavour, texture, appearance, colour, and aroma
aspects that they liked and disliked most in the samples, and they
were free to mention none or more than one attribute.
2.6. Statistical analysis
The results obtained from the different evaluations performed
in the present study were assessed using the Levene test, in order to
verify if data presented a normal distribution. ANOVA followed by
post-hoc Tukey test, set at a 5% significance level, was used when a
homogeneous variance was observed. Results displaying non-
homogeneous variance (comparison of the trials for different
time-periods studied) were evaluated using the KruskalleWallis
test for non-parametric analyses of variance, followed by Dunn's
multiple comparison. For the comparison of sampling days within a
same trial studied, the Friedman test with Bonferroni correction
was used. ANOVA for repeated measures was applied for the
evaluation of the results obtained from the sensory analysis,
whereas the Student's t-test for independent samples, set at level of
5% of significance, was used to verify differences between genders
in the product acceptability (Montgomery, 2005). Statistical soft-
wares XLSTAT 2011 (Addinsoft, USA) and MINITAB 14 (MINITAB
Inc., USA) were used.
3. Results and discussion
3.1. Dairy-based tomato spreads pH behaviour throughout storage
The results for pH values assessed in trials T1 and T3 were
similar and a significant reduction was observed after 7 and 21 days
of storage, respectively, when compared to day 1 (Table 3). Trial T2
Table 3
Values (mean ± standard deviation) of pH determined for the different trials of the
dairy-based tomato spread after 1, 7, 14, 21, and 28 days of storage at 4 ± 1
C (see
Table 1 for description of trials T1eT3).
Time (days) Trials
T1 T2 T3
1(np) 5.01 ± 0.13Aa 4.88 ± 0.06Ba 4.96 ± 0.02Aa
7 4.87 ± 0.08Ab 4.76 ± 0.07Bab 4.90 ± 0.03Aab
14 4.88 ± 0.10Ab 4.77 ± 0.09Bab 4.88 ± 0.08Aab
21 4.88 ± 0.10Ab 4.72 ± 0.05Bb 4.84 ± 0.09Abc
28 4.85 ± 0.03Ab 4.67 ± 0.10Cb 4.76 ± 0.06Bc
A,B
Different superscript capital letters in a line mean significant differences
(P < 0.05) among trials for a same day.
a,b,c
Different superscript lower case letters in a column mean significant differences
(P < 0.05) among storage days for the same trial.
(np)
Non parametric test was applied for the ready product (at the 1st day of storage).
showed lower pH values (P < 0.05) throughout the entire storage
period, when compared to T1 and T3. The addition of Lb. acidophilus
NCFM and B. animalis Bb-12 (T2) probably contributed to signifi-
cantly reduce the food product pH throughout storage, since higher
values were determined for samples containing only the starter
culture ST-M6 (T1) or ST-M6 plus Lb. sakei 2a and Bb-12 (T3). This
reduction in pH along storage observed for all trials is expected,
since a fermentative process occurs during the product
manufacturing process, in which lactose is metabolized by the
microbial cultures into lactic acid. Furthermore, the continuous
decrease in pH values of cheeses is a natural process also observed
during storage, due to the constant production of organic acids by
the starter and the probiotic cultures employed and may influence
upon the food product sensorial and technological features (Buriti,
Rocha, & Saad, 2005; Goncu & Alpkent, 2005). Similarly, Goncu and
Alpkent (2005) reported a decrease in pH of cheeses made with
yoghurt starter cultures (Lb. delbrueckii spp. bulgaricus and St.
thermophilus), cheese cultures (type O lactic culture - Lactococcus
lactis subsp. lactis and Lc. lactis subsp. cremoris) or kefir cultures
(mix of Lactobacillus spp., Lactococcus spp., Leuconostoc spp., Ace-
tobacteria spp., and yeasts) stored at 5
C during a ripening period
of 120 days. The pH decrease was also reported in Minas fresh
cheese supplemented with the probiotic Lb. acidophilus La-5 cul-
ture in co-culture with type O lactic culture (Buriti et al., 2005). It is
known that the low pH as well as other harsh features found in
some types of cheeses may negatively impact the survival of starter
and probiotic strains (Stanton et al., 1998).
3.2. Behaviour of microbial cultures in the tomato dairy-based
products throughout storage
As shown in Table 4, St. thermophilus ST-M6 populations deter-
mined in all trials were high (ca. 9 log cfu/g) and stable during the
product storage. The highest counts were observed after 21 days of
storage in T3 (9.26 log cfu/g). Lb. sakei 2a populations were also
high, as observed in T3, and remained constant throughout the food
product storage (above 8.0 log cfu/g). Lb. acidophilus NCFM counts
in T2 were significantly higher (P < 0.05) on day 7 (8.61 log cfu/g) in
comparison with those obtained on day 28 (7.93 log cfu/g).
Furthermore, probiotic Bb-12 populations were significantly higher
in T3 (P < 0.05) than those in T2, even though differences between
the two trials were below 0.5 log cfu/g, therefore of little microbi-
ological significance, and remained constant (above 8 log cfu/g)
during the storage period evaluated. The results obtained for the
viability of the probiotic cultures in the spread are in accordance
with the standards established by the Brazilian legislation, which
requires at least 8 - 9 log cfu of the microorganisms per daily
serving portion in the ready-to-eat product during its entire shelf
 C.D. Staliano et al. / LWT - Food Science and Technology 62 (2015) 682e688 685

Table 4 8e9 log cfu/mL, and also displayed high acid production,
Viability (mean values shown as log cfu/g ± standard deviation) of St. thermophilus decreasing the pH. Yoon, Woodams, and Hang (2004) studied to-
ST-M6, Lb. acidophilus NCFM, B. animalis subsp. lactis Bb-12, and Lb. sakei subsp. sakei
2a throughout storage (4 ± 1
C) assessed for the different trials of the dairy-based
mato juice fermented with Lb. acidophilus LA39 at 30
C for 72 h and
spreads evaluated (see Table 1 for description of trials T1eT3). then stored at 4
C for four weeks. The authors reported high counts
and a fast growth rate of Lb. acidophilus, achieving 9 log cfu/g and
Microorganism Time Populations of microorganisms (log cfu/g)
(days)
remaining stable, being able to survive under the adverse condi-
Trials tions of low pH and high acidity.
T1 T2 T3 Even though tomato was shown to positively influence the
St. thermophilus 1 9.35 ± 0.05Aa 9.12 ± 0.13Ba 9.17 ± 0.10Ba viability of microorganisms in the current study, different out-
ST-M6 7 9.22 ± 0.07Aab 9.21 ± 0.17Aa 9.26 ± 0.09Aa comes were observed for other vegetable-origin ingredients in
14 9.12 ± 0.07ABab 9.07 ± 0.11Ba 9.20 ± 0.13Aa probiotic products. Espírito Santo, Perego, Converti, and Oliveira
21 9.03 ± 0.18Bab 9.04 ± 0.24Ba 9.26 ± 0.17Aa (2012) added passion fruit peel powder in skim yoghurt supple-
28 8.95 ± 0.36Bb 9.00 ± 0.50Ba 9.24 ± 0.09Aa
Lb. acidophilus 1 e 8.50 ± 0.16ab e
mented with Lb. acidophilus NCFM. The authors observed that the
NCFM 7 e 8.61 ± 0.23a e passion fruit led to an inhibitory effect of the probiotic strain, since
14 e 8.30 ± 0.36ab e populations were significantly reduced throughout 28 days of
21 e 8.16 ± 0.46ab e storage. Also, in a study with milk-based mousses (Buriti, Komatsu,
28 e 7.93 ± 0.93b e
& Saad, 2007b), the addition of passion fruit as concentrated juice
B. animalis subsp. 1 e 8.43 ± 0.19Ba 8.62 ± 0.11Aa
lactis Bb-12 7 e 8.41 ± 0.04Bab 8.60 ± 0.16Aa or pasteurized frozen pulp reduced Lb. acidophilus La-5 viability in
14 e 8.34 ± 0.06Babc 8.56 ± 0.09Aa 4.7 log cycles after 21 days of storage at 4
C. Different results were
21 e 8.25 ± 0.07Bc 8.58 ± 0.06Aa reported for fermented soy products, in which the addition of
28 e 8.26 ± 0.11Bbc 8.54 ± 0.12Aa mango and guava pulps did not affect Lb. acidophilus La-5 and
Lb. sakei subsp. 1 e e 8.18 ± 0.20a
Bifidobacterium Bb-12 viability (Bedani, Vieira, Rossi, & Saad, 2014).
sakei 2a 7 e e 8.17 ± 0.20a
14 e e 8.10 ± 0.14a Lb. acidophilus NCFM and Lb. sakei 2a showed high and stable
21 e e 8.16 ± 0.08a viability in the dairy matrix studied throughout storage of T2 and
28 e e 8.11 ± 0.11a T3, respectively. Both LAB displayed a good interaction with the
A,B
Superscript capital letters, in a line, mean significant differences (P < 0.05) among probiotic strain Bb-12, as observed by the high counts of the
trials for a same day studied. microorganism (around 8 log cfu/g).
a,b,c
Superscript lower case letters, in a column, mean significant differences The pH values determined in T2 were statistically lower
(P < 0.05) among days for the same trial and microorganism evaluated.
(P < 0.05) than those assessed for T3 and T1 (Table 3), probably due
to the higher ability of NCFM to degrade lactose present in milk into
lactic acid. In reality, Lb. sakei strains are frequently used as starter
life (ANVISA, 2008). It is worthy to mention that, for similar types of cultures in fermented meat products, including sausages, due to
spread, Brazilian standards recommend a daily serving portion of their ability to quickly ferment carbohydrates (Champomier-
10 g (ANVISA, 2003). Therefore, the tomato-based dairy spread was Verge s, Chaillou, Cornet, & Zagorec, 2001). Lb. sakei 2a is a poten-
shown to be a good vehicle for the delivery of LAB and Bb-12. tially probiotic strain that was originally isolated from a meat
St. thermophilus easily adapted to the dairy product, since it is a product with very different intrinsic characteristics. However, we
homofermentative LAB commonly used as starter culture in yo- can clearly infer that the microorganism showed a good adaptation
ghurts (together with Lb. delbrueckii subsp. bulgaricus) and fer- to the dairy food matrix tested. Likely, the promising results ob-
mented milk (Saad, Komatsu, Granato, Branco, & Buriti, 2011). The tained from this study suggest that the microorganism could also
addition of St. thermophilus ST-M6 was an interesting approach, be applied as a starter culture in fermented dairy products. Even
since it remained viable during the product storage and it is
required for milk acidification to promote coagulation through the
action of chymosin on the k-casein steric stabilizing layer of the
casein micellae, which is needed to obtain cheese (Cruz et al.,
2009).
There are many studies using St. thermophilus as a starter culture
and supplemented with probiotics. Among them, Buriti, Cardarelli,
and Saad (2007) used St. thermophilus in co-culture with Lb. para-
casei to produce potentially probiotic and synbiotic fresh cream
cheeses. The authors reported St. thermophilus populations above
9.5 log cfu/g for both kinds of cheeses during storage for up to 21
days. In another study, St. thermophilus and Lb. acidophilus NCFM
were added in co-culture in yoghurt stored during 28 days at 4
C,
and the authors observed an adequate probiotic viability
(0.5 log cfu/g reduction) (Ng, Yeung, & Tong, 2011).
As shown by our data, it is important to stress out that the as-
sociation of milk and tomato in the dairy-based spread was bene-
ficial for an adequate adaptation and high viability rates of Lb.
acidophilus NCFM. Similarly, some studies have also demonstrated a
positive adaptation of Lactobacillus spp., particularly Lb. acidophilus,
to the dairy matrix of food products with tomato. Babu, Mital, and
Grag (1992) evaluated the addition of tomato juice in skimmed
Fig. 1. Sensory scores attributed by female ( ) and male ( ) consumers for the
milk inoculated with different strains of L. acidophilus. After incu- different trials of the dairy-based tomato spreads stored for 7, 14, and 21 days at
bation at 37
C for 16e18 h, the authors observed that the micro- 4 ± 1
C. Bars mean the average values given by each gender displaying two units of
organism was able to grow, exhibiting final populations of standard deviation (see Table 1 for description of trials T1eT3).
686 C.D. Staliano et al. / LWT - Food Science and Technology 62 (2015) 682e688

though Lb sakei 2a is believed to produce bacteriocins in the food
matrix tested, which apparently did not affect the high viability of
ST-M6 and Bb-12 throughout storage, a more detailed inquiry is
currently being undertaken in our laboratory to verify this
possibility.
Several studies evaluated the use of Lactobacillus in co-culture
with Bifidobacterium in food systems. Pereira, Souza, Behrens, and
Saad (2010) added Lb. acidophilus La-5 and B. animalis subsp. lac-
tis BL04 to petit-suisse cheese formulations and observed pop-
ulations above, respectively, 7 log cfu/g and 8 log cfu/g, throughout
28 days of refrigerated storage. As demonstrated in the present
study, Bb-12 viability was also high (Table 4) in T2 and T3.
Even though all the products in this study included the prebiotic
fibre inulin in their formulation, it is possible that the compound
contributed for the high viability of the microbial populations
determined during the dairy product storage period, as observed by
Oliveira, Perego, Oliveira, and Converti (2011). The authors
demonstrated that the supplementation of inulin in skimmed milk
fermented with a cocktail containing Lb. acidophilus, Lb. rhamnosus,
Lb. bulgaricus, Bifidobacterium lactis, and St. thermophilus led to
significantly lower (17e48%) generation times compared to those
obtained in pure skim milk. The authors attributed these results to
a positive prebiotic effect, which improved the probiotic strains
growth rates.
3.3. Sensory evaluation of the dairy-based tomato spreads during
storage
In the present study, an affective sensory test was included to
determine if consumers liked or not (and at what intensity) the

A 45.0%

40.0%

35.0%

30.0%
Frequency

25.0%

20.0%

15.0%

10.0%

5.0%

0.0%
1 2 3 4 5 6 7 8 9
Scores

B 45.0%
40.0%
35.0%

30.0%
Frequency

25.0%
20.0%
15.0%
10.0%
5.0%
0.0%
1 2 3 4 5 6 7 8 9
Scores

C 45.0%
40.0%
35.0%
30.0%
Frequency

25.0%
20.0%
15.0%
10.0%
5.0%
0.0%
1 2 3 4 5 6 7 8 9

Fig. 2. Frequency (expressed as percentage) of scores (ranging from 1 to 9) attributed by consumers for the different trials ( ) of the dairy-based tomato spreads stored
for 7 (A), 14 (B), and 21 (C) days at 4 ± 1
C (see Table 1 for description of trials T1eT3).
 C.D. Staliano et al. / LWT - Food Science and Technology 62 (2015) 682e688
innovative food product developed. According to Cruz, Cadena,
Farias, and Bolini (2011), this is considered a chief assessment in
order to determine the potential for commercialization of new food
products supplemented with pro- and/or prebiotics, which is the
case of our potentially synbiotic dairy-based tomato spread.
Fig. 1 summarizes the results obtained for the sensory accept-
ability test conducted for T1, T2, and T3. The mean overall scores
were given by different genders of consumers, of which 55.26%
were women and 44.74% men. There was not a significant differ-
ence (P > 0.05) among the scores given by the consumers for any
product during the different periods evaluated for the three trials.
The mean scores attributed for the samples ranged from 6.04
(minimum value given by a male consumer for T1 on day 14) to 8.30
(maximum rate given by a male consumer for T2 on day 14). Also,
there was not a remarkable difference between the minimum and
the maximum average scores judged by the consumers, which is a
positive outcome and demonstrates that the sensory characteristics
of the dairy-based tomato spread were satisfactory. The mean
scores for all samples were higher among the female consumers in
seven out of nine sections, in comparison with those given by the
male consumers; however, no significant difference (P > 0.05) was
observed among them. The mean scores were at least 7 (liked
regularly) considering all trials, with a high frequency of scores 7, 8,
and 9 (Fig. 2), which demonstrates that the innovative product was
well accepted by consumers since the product global impression
was quite good and adequate balance of the ingredients, colour, and
texture were features frequently mentioned by them. Taking into
account that the development of new functional products has
resulted in an exponential growth of the probiotic market in the
last five years, our positive findings indicate good perspectives for
the dairy-based tomato spread commercialization.
Similar positive results were observed by Tsen, Lin, Huang, and
King (2008), who developed a tomato juice fermented with Lb.
acidophilus immobilized in k-carrageenan. The authors observed a
higher sensorial acceptability, when compared to the fermented
juice containing free cells of the microorganism. Moreover, the
scores related to palatability were higher at the beginning of stor-
age at 4
C, remaining constant up to 10 weeks. Antunes et al.
(2009) evaluated the acceptability of a probiotic buttermilk con-
taining Bb-12, with different flavours (strawberry, vanilla, mint,
soursop, and cupuassu), containing either sucrose or sucralose. The
authors observed scores ranging from 6 to 8 for all samples and
concluded the results were all satisfactory.
The role played by the starter cultures is important since they
may contribute for the development of flavour in cheeses during
storage, as a consequence of their proteolytic action and carbohy-
drates metabolism (Candioti et al., 2002). Moreover, the addition of
Bb-12 together with Lb. acidophilus NCFM or Lb. sakei 2a in the
dairy-based spread tested in here did not impair its overall
acceptance.
These results are important to improve the current worldwide
scenario, where a higher development and consumption of func-
tional foods is being observed, together with the concern of con-
sumers for healthier eating habits and food re-education associated
with health and welfare. Also, the manufacturers of functional food
products should be aware of the importance to provide full de-
scriptions of the probiotic strain(s) and prebiotic(s) employed on
the food products' label in order to claim the accurate healthy
benefits associated with their regular consumption.
4. Conclusion
The present study showed that the innovative functional dairy-
based tomato spread containing prebiotic fibre and probiotic and
potentially probiotic microorganisms plus the antioxidant lycopene
687
(present in the dried tomato and in tomato pulp added as in-
gredients) was technologically feasible and exhibited high viability
results for the beneficial microorganisms Lb. acidophilus NCFM and
Bb-12 during the entire period of storage tested. Additionally, Lb.
sakei subsp. sakei 2a, a bacteriocinogenic and potentially probiotic
strain, showed high counts throughout the product storage and
could, therefore, be adequately incorporated into the product dairy
matrix. Sensory evaluation revealed mean scores higher than 7
(liked regularly) for all trials, which demonstrated the product was
well accepted by consumers. Moreover, future researches should be
done in order to investigate the synbiotic potential of the product
through in vitro tests and clinical trials.
Acknowledgements
The authors thank “Fundaç~ ao de Amparo a  Pesquisa do Estado
de Sa~o Paulo” (FAPESP) (Projects # 2009/13347-3, 2009/52600-6,
and 2010/52267-2) and “Coordenaça ~o de Aperfeiçoamento de
Pessoal de Nível Superior” (CAPES) for financial support and fel-
lowships. The authors also thank Danisco, Orafti, and Salute com-
panies for the donation of some material sources, and Prof.
Bernadette D. G. M. Franco for providing the Lb. sakei subsp. sakei 2a
strain.
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