Journal of Food Engineering 99 (2010) 128–135

Contents lists available at ScienceDirect

Journal of Food Engineering

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

Rheological and physico-chemical characterization of prebiotic microfiltered

fermented milk
Janaina Debon a, Elane Schwinden Prudêncio b,*, José Carlos Cunha Petrus a
a
Departamento de Engenharia Química e Engenharia de Alimentos, Centro Tecnológico, Universidade Federal de Santa Catarina, Trindade, 88040-970, Florianópolis, SC, Brazil
b
Departamento de Ciência e Tecnologia de Alimentos, Centro de Ciências Agrárias, Universidade Federal de Santa Catarina, Rod. Admar Gonzaga, 1346,
Itacorubi, 88034-001, Florianópolis, SC, Brazil

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

Article history: Microfiltered cow’s milk samples were fermented with (prebiotic) and without inulin and evaluated
Received 2 September 2009 regarding the physico-chemical composition (day 1 of storage) and rheological (days 1, 7, 14 and 21)
Received in revised form 20 January 2010 evaluation, at temperatures of 4.0 ± 0.1 °C and 6.0 ± 0.1 °C. The addition of inulin increased the total solids
Accepted 8 February 2010
content of the prebiotic fermented milk, increasing its acidity. The Power Law and Mizhari and Berk mod-
Available online 16 February 2010
els were applied successfully to describe the flow of the fermented milks, which had characteristics of
shear thinning and non-Newtonian fluid behavior. As with the addition of inulin, the storage time con-
Keywords:
tributed to an increase in the apparent viscosity of the product. The prebiotic fermented milk had greater
Microfiltration
Fermented milk
thixotropy and the hysteresis was lower with an increase in the temperature.
Prebiotic Ó 2010 Elsevier Ltd. All rights reserved.
Inulin
Rheology

1. Introduction and Meyer, 2006). Inulin is a non-digestible ingredient which

One of the applications of microfiltration in dairy product plants
is the retention of the milk bacteria and spores (Lawrence et al.,
2008; Coutinho et al., 2009), aiming to obtain a permeate (Saboya
and Maubois, 2000) which can be employed in the production of
milk derivatives, such as fermented milks.
The use of microfiltration to replace traditional thermal pro-
cesses, such as pasteurization, can reduce changes in the chemical
properties of the milk, such as the protein denaturation (Tziboula
et al., 1998; Mistry and Moubois, 2004). However, it is fundamen-
tal to skim the milk before microfiltration, so that good permeate
flux can be obtained. Since the fat globules are larger than the
microfiltration membrane pores, they are rejected and tend to
form a gel layer on its surface. This phenomenon leads to an impor-
tant reduction in permeate flux, besides making the cleaning of the
membrane difficult (Saboya and Maubois, 2000). Therefore, the
production of fermented products from microfiltered milk, without
fat and cold pasteurized, may result in a product with different
properties, mainly regarding the rheological behavior.
Modifications in the rheological properties of fermented milks,
can be observed when functional food ingredients are incorpo-
rated, for instance, inulin (Hauly and Moscatto, 2002; Tungland
* Corresponding author. Tel.: +55 48 37215366; fax: +55 48 37219943.
E-mail address: elane@cca.ufsc.br (E.S. Prudêncio).
beneficially affects humans, since it selectively stimulates the
growth and/or activity of one or a limited number of bacteria in
the colon (Gibson and Roberfroid, 1995). Furthermore, inulin is sta-
ble with moderate acidity (Martínez-Villaluenga et al., 2005), its
use being recommended in the production of fermented milk
derivatives. Technologically, inulin has been applied as a fat substi-
tute in some foods with the advantage of not increasing its calorific
value and not giving a residual taste to the product (Flamm et al.,
2001; Roberfroid, 2005). Besides the selective stimulation of bacte-
ria, Gibson et al. (2004) reported that these ingredients are classi-
fied as prebiotic because they resist the processes of digestion,
absorption and adsorption in the gastrointestinal tract.
Recent studies have shown that the changes in foods with inulin
addition favorably affect the acceptance of the product by the con-
sumer (Mohameed et al., 2004). These modifications also influence
the engineering calculations of the process, such as the flow, selec-
tion of pumps, determination of load loss in the tubes, etc. (Holds-
worth, 1993; Steffe, 1996; Tabilo-Munizaga and Barbosa-Cánovas,
2005), as well as in the sizing of equipment (Maskan and Gogus,
2000; Abu-Jdayil and Mohameed, 2002).
As verified by Ziemer and Gibson (1998), information related to
the behavior of prebiotics in fermented and stored milk products is
practically inexistent. Thus, the objective of this study was to use
the microfiltrate of cow’s milk, with acceptable microbiological
characteristics, to produce a prebiotic fermented milk product,

0260-8774/$ - see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.jfoodeng.2010.02.008
 J. Debon et al. / Journal of Food Engineering 99 (2010) 128–135
Nomenclature
g apparent viscosity
r shear stress, Pa
K consistency index, Pa s1
KM consistency index, Pa1/2 sn
with suitable physico-chemical composition. The influence of the
inulin addition on the rheological properties during 21 days of
storage was evaluated.
2. Material and methods
2.1. Material
Raw skimmed milk, inulin (RaftilineÒ HP-Gel, Beneo, Orafti, Or-
eye, Belgium), thermophylic milk culture (YC-X11 Yo FlexÒ, Chr.
Hansen) composed of Lactobacillus delbrueckii ssp. bulgaricus and
Streptococcus salivarius ssp. thermophilus, and saccharose were
used. All reagents were of a high analytical grade.
2.2. Microfiltration (MF)
The raw skimmed milk was microfiltered in a pilot unit, using
an organic poly(imide) membrane, hollow fiber type, manufac-
tured by PAM Membranas Seletivas, Rio de Janeiro, Brazil, with
an average pore diameter of the order of 1.4 lm, and filter area
of 0.4 m2. The operating parameters controlled during the microfil-
tration process were temperature of 45 ± 1 °C, pressure of 3 bar
and tangential velocity of 0.8 m s1, up to a VRF (Volumetric
Reduction Factor) of 4. During the microfiltration an average per-
meate flux of 41.27 L h1 m2 was obtained. After each processing
the pilot unit and membrane were cleaned and hygienized, accord-
ing to the manufacturer’s instructions.
2.3. Production of fermented milks
For the production of the fermented milk from the microfiltrate,
the method proposed by Almeida et al. (2001) was adapted. The
milk, heated and maintained at 45 ± 1 °C, was microfiltered and
used to produce the prebiotic fermented milk, with the addition
of 5% inulin, as recommended by Jelen and Lutz (1998), Niness
(1999) and Roberfroid (1999). Fermented milk, without inulin
addition, was also produced as a control. To both products 8% of
saccharose was added, as well as the thermophylic milk culture,
inoculated at a temperature of 42 ± 1 °C. The products were fer-
mented up to pH 4.5 (±0.2) and immediately cooled to 10 °C, gently
stirred and was put into polypropylene cups sealed and stored 5 °C
(±1 °C), until the carrying out of the physico-chemical analysis on
day 1 of storage, and rheological analysis on days 1, 7, 14 and 21.
2.4. Microbiological analysis
The counts of mesophilic and psychrophilic bacteria were taken
according to the methodology described in APHA (2001). The re-
sults were expressed as colony-forming units per millilitre (cfu/
mL) of product. This analysis was carried out in triplicate.
2.5. Physico-chemical analysis
The raw skimmed milk, the microfiltrate and fermented milks
(prebiotic and control) were analyzed for total solids content (%
m/m), through the drying of the samples until constant weight
(925.23) (AOAC, 2005). The acidity (% lactic acid) of the fermented
129
KOM square root of the yield stress, Pa1/2
c_ shear rate, s1
N flow behavior index
nM flow behavior index
milks was determined according to the method of Instituto Adolfo
Lutz (IAL, 2005), while the pH values were obtained using a pH me-
ter (±0.01) (MP220, Metler-Toledo, Greinfensee, Switzerland). All
of these analyses were carried out in triplicate.
2.6. Rheological measurements
The measurements were carried out using a Thermo Haake DC
10 rotational viscosimeter (model VT 550, Thermo Haake, Kar-
lsruhe, Germany), with concentric cylinders (NV ST 807-0713 CE
and NV 807-0702), and collected using the software program Pro
RheowinÒ (version 2.93, Haake). The flow curves were generated
by shear rate increased linear from 10.82 s1 to 221.80 s1 in the
first 20 min (upward curve) and returned to 10.82 s1 in the follow-
ing 20 min (downward curve), under controlled temperature
4.0 ± 0.1 °C and 6.0 ± 0.1 °C, through water circulation in a temper-
ature controlled bath (Phoenix P1, Thermo Haake, Karlsruhe, Ger-
man) and coupled to the equipment. The rotational speed was
increased from 2 rpm to 41 rpm, at 2 rpm per minute. The flow
behavior was described by the Power Law model (r = K(c_ )n, where
r is shear stress (Pa), c_ is shear rate (s1), K is consistency index
(Pa s1) and n is flow behavior index) and by the Mizrahi and Berk
model (r½ = KOM + KMðc_ ÞnM nM , where r is shear stress (Pa), KOM is
square root of the yield stress (Pa1/2), KM is consistency index
(Pa1/2 sn), c_ is shear rate (s1), and nM is flow behavior index). Vis-
cosity values on the downward (viscosity/shear rate) curves at a
rate of 50 s1 were considered as the apparent viscosity (g) of the
fermented milk samples, which according to Bourne (2002), repre-
sent the approximate viscosity perceived on the palate. The thixo-
tropic behavior of the fermented milks was evaluated by
calculating the hysteresis loop area between the upward and down-
ward flows curves. In order to determine the thixotropy and appar-
ent viscosity, the measurements were carried out at controlled
temperatures (4.0 ± 0.1 °C and 6.0 ± 0.1 °C). The rheological param-
eters K, KM, KOM, n, nM, g and the flow curves (shear stress versus
shear rate) were obtained considering an initial shear rate of 40
s1. All of the values were obtained in duplicate.
2.7. Statistical analysis
The data were expressed as means and standard deviation. One-
way analyses of variance (ANOVA) and Tuckey’s studentized range
(5% significance) were carried out to test any significant differences
between the results. The data were obtained using the software
STATISTICA version 6.0 (2001) (StatSoft Inc., Tulsa, OK, USA).
3. Results and discussion
3.1. Microbiological analysis
The microfiltrate had a mesophile count of 3 log cfu mL1 and
psychrophile count of 2 log cfu mL1. These values were lower
and similar to those obtained by Zocche et al. (2002), for the mes-
ophile (between 3 and 4 log cfu mL1) and psychrophile (between
2 and 3 log cfu mL1) counts, respectively, while Pietrowski et al.
(2008) reported mesophile counts of between 2 and 4 log cfu mL1,
in pasteurized milks.
130 J. Debon et al. / Journal of Food Engineering 99 (2010) 128–135

Figueiredo and Porto (2002) reported that high mesophilic bac- 1600
teria counts lead to a greater lactic acid production, resulting in Control
more acidic yogurts, adversely affecting their physical, chemical, 1400
(a) Prebiotic
sensorial and rheological qualities. Values for psychrophilic micro-
1200
organism counts of 4 log cfu mL1 (Adams et al., 1975), between 5
and 6 log cfu mL1 (Muir, 1996) and 6 and 7 log cfu mL1 (Schröder 1000
and Bland, 1983), result in the production of thermostable en-

Viscosity (mPa.s)
zymes (proteolitics and lipolitics) which can degrade the milk 800
components, produce disagreeable odor and taste, and influence
the coagulation and the yield of milk derivatives. Also, Figueiredo 600
and Porto (2002) reported that microbial enzymes reduce the yo-
400
gurt quality, influencing its whey retention capacity by the protein
network and also its viscosity.
200
Smithwell and Kailasapathy (1995) verified that milk contami-
nation by psychrophiles is a problem which affects the whole dairy 0
industry, being considered the most important factor in the control 0 30 60 90 120 150 180 210 240
of the maintenance of milk quality aiming to increase its shelf life. Shear rate (s -1)
Finally, the low counts of mesophilic and psychrophilic micro-
organisms observed in the microfiltrate demonstrate that it has 1600
suitable microbiological quality, and can thus be employed in the
Control
production of prebiotic fermented milk. 1400
(b) Prebiotic

3.2. Physico-chemical analysis 1200

1000
The average values for the physico-chemical properties of raw
Viscosity (mPa.s)

skim milk, microfiltrate and fermented milks are shown in Table 1.
As expected, the addition of inulin to prebiotic fermented milk
resulted in an increase (p < 0.05) of total solids content. The same
behavior was observed by Akalin et al. (2007) and Guggisberg
et al. (2009) for yogurt supplemented with inulin, and also by Cas-
tro et al. (2008) and Villegas et al. (2009) in fermented milk with
oligofructose and inulin addition, respectively. As verified by
Wolfschoon-Pombo et al. (1983), in yogurts, a relation between
the increase in total solids and acidity (p < 0.05) was also observed
in prebiotic fermented milk. According to Martínez-Villaluenga
et al. (2005) these results may also be related to the use of inulin
to initiate the cultures during the fermentation, which would lead
to acid formation.
No differences were observed between the pH values (p < 0.05)
of the fermented milks (control and prebiotic). Guven et al. (2005)
and Guggisberg et al. (2009) also reported that the addition of inu-
lin did not alter the pH of fermented milks. The pH values obtained
were in agreement with those suggested by Tamime and Robinson
(2007) and Gallardo-Escamilla et al. (2007), who recommend as
desirable the obtainment of fermented milks with pH between
4.2 (prebiotic) and 4.8 (control). Tamime and Robinson (2007) re-
lated that the obtainment of fermented milks in this pH range
would results in a better rearrangement and aggregation of casein
particles, contributing according to Gallardo-Escamilla et al. (2007)
to the formation of a more stable gel, avoiding the separation of
phases, i.e., syneresis.
3.3. Rheological measurements
The apparent viscosity of the control and prebiotic fermented
milk samples, decreased with an increase in the shear rate, indicat-
800
600
400
200
0
0 30 60 90 120 150 180 210 240
Shear rate (s -1)
Fig. 1. Apparent viscosity versus shear rate at temperatures of (a) 4.0 ± 0.1 °C and
(b) 6.0 ± 0.1 °C for control and prebiotic fermented milks.
ing a non-Newtonian fluid behavior (Fig. 1a and b). The decrease in
the apparent viscosity of the fermented milks with the increase in
shear rate at the temperatures evaluated indicates that the fluid
has shear thinning characteristics, being in agreement with results
obtained by Donkor et al. (2007) and Guggisberg et al. (2009) for
yogurt with inulin addition; by Castro et al. (2008) for a symbiotic
lactic beverage; and by Cunha et al. (2008) for probiotic fermented
milks. The reduction in the apparent viscosity with the increase in
shear rate, in these types of products, may be related to several fac-
tors. Lucey (2002) suggests that this results from the destruction of
weak bonds and a decrease in the sum of the electrostatic repul-
sion and hydrophobic interaction between the molecules of the
gel. Karazhiyan et al. (2009) reported that the increase in shear rate
and decrease in viscosity occur due to a breaking of the macromo-
lecular structures in solution, due to the shear force. Alparslan and
Hayta (2002) and Castro (2003) reported that the hydrodynamic
forces, at the beginning of shearing, are more intense, causing

Table 1
Results for the physico-chemical analysis of the raw skimmed milk ± standard deviation, of the microfiltrate and of the fermented milks (control and prebiotic).

Total solids (% m/m)* Acidity (% lactic acid)* pH*

Raw skim milk 8.80 ± 0.02 0.19 ± 0.01 6.66 ± 0.00
Microfiltrate 8.41 ± 0.03 0.18 ± 0.01 6.67 ± 0.00
Control fermented milk 14.50a ± 0.03 1.67a ± 0.01 4.27a ± 0.01
Prebiotic fermented milk 18.39b ± 0.08 1.71b ± 0.01 4.28a ± 0.02
*
Results expressed as mean ± standard deviation. Different letters in the same column indicate significant differences between samples (p < 0.05).
 J. Debon et al. / Journal of Food Engineering 99 (2010) 128–135 131

Table 2
Rheological parameters obtained using the Power Law model (g = K(c_ )n1) for control and prebiotic fermented milks at temperatures of 4.0 ± 0.1 °C and 6.0 ± 0.1 °C, on storage
days 1, 7, 14 and 21.

Sample T (°C) Day K* (Pa sn) n** g*** (mPa s) R****

Aa
Control 4 1 0.213 0.650 54.21 0.996
7 0.197 0.696 59.97Ab 0.997
14 0.185 0.704 58.14Ab 0.996
21 0.228 0.655 59.21Ab 0.995
Control 6 1 0.215 0.634 51.29Aa 0.996
7 0.215 0.664 57.77Ab 0.996
14 0.219 0.657 57.11Ab 0.995
21 0.222 0.646 55.13Ab 0.995

Prebiotic 4 1 0.132 0.816 64.41Ba 0.994

7 0.139 0.809 65.44Ba 0.995
14 0.190 0.758 73.92Bb 0.996
21 0.187 0.753 71.13Bb 0.994
Prebiotic 6 1 0.117 0.821 58.29Ba 0.995
7 0.136 0.805 63.73Ba 0.995
14 0.209 0.732 72.94Bb 0.994
21 0.219 0.706 71.96Bb 0.994
A,B
Values followed by different letters in the same column indicate significant differences (significance level of 5%) between the formulations on the same storage day and in
the same temperature. a,b,cValues followed by different letters in the same column indicate significant differences (significance level of 5%) between the storage days for same
sample and temperature.
*
Consistency index (K).
**
Flow behavior index (n).
***
Apparent viscosity (g) a c_ de 50 s1.
****
Correlation coefficient (R).

Table 3
Rheological parameters obtained using the Mizrahi and Berk model (g = ((KOM + KM(c_ )nM)2)/c_ ) for control and prebiotic fermented milks at temperatures of 4.0 ± 0.1 °C and
6.0 ± 0.1 °C, on storage days 1, 7, 14 and 21.

Sample T (°C) Day KOM* (Pa) KM** (Pa s) nM*** g**** (mPa s) R*****
Aa
Control 4 1 0.852 0.091 0.551 55.50 0.998
7 0.924 0.080 0.593 61.77Ab 0.999
14 0.985 0.063 0.638 60.24Ab 0.998
21 1.101 0.048 0.662 61.33Ab 0.999
Control 6 1 0.92 0.072 0.583 52.65Aa 0.998
7 1.020 0.061 0.629 59.66Ab 0.998
14 1.019 0.059 0.629 58.62Ab 0.998
21 1.017 0.057 0.627 56.86Ab 0.998

Prebiotic 4 1 1.257 0.023 0.838 69.30Ba 0.999

7 1.207 0.029 0.794 69.84Ba 0.999
14 1.436 0.018 0.867 79.30Bb 0.999
21 1.295 0.030 0.782 75.35Bb 0.998
Prebiotic 6 1 1.149 0.026 0.808 62.48Ba 0.999
7 1.131 0.037 0.755 67.57Ba 0.998
14 1.345 0.028 0.791 77.18Bb 0.998
21 1.393 0.024 0.803 75.86Bb 0.999
A,B
Values followed by different letters in the same column indicate significant differences (significance level of 5%) between the formulations on the same storage day and in
the same temperature. a.b.cValues followed by different letters in the same column indicate significant differences (significance level of 5%) between the storage days for same
sample and temperature.
*
Square root of the yield stress (KOM).
**
Consistency index (KM).
***
Flow behavior index (nM).
****
Apparent viscosity (g) a c_ de 50 s1.
*****
Correlation coefficient (R).

greater rupture. Over time, these forces generate system stretching
allowing the alignment with the flow and, consequently, a reduc-
tion in the viscosity values.
In Fig. 1a and b, at the two temperatures evaluated, it can be
verified that the prebiotic fermented milk, had a higher initial vis-
cosity, with a steep decrease with the increase in the shear rate.
This behavior was to be expected since, according to Guggisberg
et al. (2009) the use of inulin at concentrations greater than 1% re-
sults in more viscous products. Toneli et al. (2007), besides noting
the ability of inulin to form a gel, reported that it can also increase
the viscosity of yogurts. Moreover, González-Tomás et al. (2008)
verified that despite being more thermally stable than other types
of fibers, inulin tends to form microcrystals in milk, which retain a
great quantity of water, resulting in a more viscous gel. This behav-
ior is associated by Silva (1996) with the capacity of one molecule
of inulin to bind to two molecules of water.
The rheological parameters evaluated of the fermented milks
evaluated during 21 days of storage are given in Tables 2 and 3.
The values for the correlation coefficients (R), for the Power Law
and Mizrahi and Berk models, were equal to or greater than
132 J. Debon et al. / Journal of Food Engineering 99 (2010) 128–135

12 12
(a) (b)
10 10
Shear stress (Pa)

Shear stress (Pa)

8 8

6 6

4 4 Control T = 4°C
Control T = 4°C
Control T = 6°C Control T = 6°C
Prebiotic T = 4°C Prebiotic T = 4°C
2 2
Prebiotic T = 6°C Prebiotic T = 6°C

40 60 80 100 120 140 160 180 200 220 240 40 60 80 100 120 140 160 180 200 220 240

Shear rate (s-1) Shear rate (s- 1)

12 12
(c) (d)
10 10

Shear stress (Pa)

Shear stress (Pa)

8 8

6 6

4 4 Control T = 4°C
Control T = 4°C
Control T = 6°C Control T = 6°C
Prebiotic T = 4°C Prebiotic T = 6°C
2 2
Prebiotic T = 6°C Prebiotic T = 6°C

40 60 80 100 120 140 160 180 200 220 240 40 60 80 100 120 140 160 180 200 220 240

Shear rate (s- 1) Shear rate (s- 1)

Fig. 2. Flow curves, shear stress versus shear rate, for control and prebiotic fermented milks at temperatures of 4.0 ± 0.1 °C and 6.0 ± 0.1 °C, on storage days 1 (a), 7 (b), 14 (c)
and 21 (d).

0.994. Thus, it was verified that both of the models were appropri-
ately employed, that is, they described the rheological behavior of
the fermented milks, for the different storage times and tempera-
tures used.
At the storage times and temperatures evaluated, the flow
behavior index (n) of the fermented milks, obtained through the
both models (Tables 2 and 3), showed characteristics of a shear
thinning fluid (n < 1), thus confirming the non-Newtonian
behavior.
As verified by Fuchs et al. (2006) in the obtainment of a yogurt
supplement with inulin; by Bortolozo and Quadros (2007) in the
application of inulin and sucralose in yogurts; and by Donkor
et al. (2007) and Guggisberg et al. (2009) in yogurts with inulin
added, the prebiotic fermented milk developed in this study had
a greater apparent viscosity (p < 0.05) than the control, during
the 21 days of storage, at the temperatures evaluated (Tables 2
and 3). This increase can be explained through the work of Akin
et al. (2007) and Soukoulis et al. (2009) who reported that inulin
(soluble fiber) contributes to a modification in the physical proper-
ties of foods, due to its capacity to retain water, aiding the forma-
tion of viscous gels. Gonzalez-Tomás et al. (2008) reported that
these rheological changes caused by the inulin are due to its struc-
ture being composed of a network of microcrystals, formed by
small aggregates, able to retain water. Schaller-Povolny and Smith
(2001) reported that when inulin binds to the water molecules it is
observed that these also interact with the milk protein, leading to
an increase in the molar mass, which results in an increase in the
viscosity. Furthermore, Kip et al. (2005) and Villegas and Costell
(2007) verified that the addition of greater quantities of inulin to
yogurts also increased the apparent viscosity of the product, con-
firming the findings of Brink et al. (2007) and Teles and Flôres
(2007) who associated this increase with a greater total solids con-
tent, as verified in this study (Table 1).
The storage time of the fermented milks (control and prebiotic)
influenced significantly their apparent viscosity (Tables 2 and 3). In
the control fermented milk the difference in the apparent viscosity
(p < 0.05) was observed from day 7, while for the prebiotic fer-
mented milk the increase (p < 0.05) was verified from day 14. How-
ever, from these periods up to day 21 of storage the g values
remained constant (p > 0.05). The same behavior was observed
by Cunha-Neto et al. (2005) and Cueva et al. (2008) for yogurts
stored for 30 and 21 days, respectively. In a similar way to that ver-
ified by Donkor et al. (2007) for yogurts with (between 0.5% and
1.5%) or without inulin, stored for 28 days, in the fermented milks
(control and prebiotic) an increase (p < 0.05) in the viscosity was
also observed with the period of storage.
Abu-Jdayil and Mohameed (2002) also observed an increase in
the viscosity of labneh with a storage time of 14 days. According
to Ozer et al. (1998) the greater number of interactions and, conse-
quently, the rearrangement of proteins would be responsible for
the increase in viscosity. Since Ross-Murphy (1990) states that this
behavior is common in acid gels during storage.
The rheological evaluation carried out for 21 days of storage
showed that the temperature variation of 4.0 ± 0.1 °C to
6.0 ± 0.1 °C reduced the apparent viscosity (p < 0.05) of the fer-
mented milks (control and prebiotic). As predicted, this behavior

400 383.8a
a
350
338.1
Hysteresis loop area (unit)
300.9b
300
251.2a
250
b
236.2
200
150
100
50
0
1 2 3 4
Storage (days)
Fig. 3. Hysteresis area for control fermented milk, at temperatures of 4.0 ± 0.1 °C
(h) and 6.0 ± 0.1 °C ( ) on storage days 1, 7, 14 and 21.
is in agreement with Machado (2002) who verified the relation be-
tween the temperature and the viscosity reduction. The decrease in
apparent viscosity with an increase in temperature, according to
Cruz et al. (2008), is related to the increase in thermal energy,
which reduces the intermolecular interaction, while Hassan and
Hobani (1998) and Cruz et al. (2008) state that this low interaction
results in smaller sized particles, facilitating the flow and reducing
the viscosity of the product. Karazhiyan et al. (2009) reported that
this behavior may also occur due to the increase in the mobility of
the macromolecules, indicating less resistance to flow.
Through the rheograms shown in Fig. 2a–d it can be observed
that both the control and prebiotic fermented milks show hystere-
sis, indicating, in the same way as Tárrega et al. (2004) and Toneli
et al. (2005), that this is dependent on storage time. Toneli et al.
(2005) reported that the formation of a hysteresis curve reflects
changes in the rheological behavior of the product.
The same behavior observed in this study for prebiotic fer-
mented milk, which had a higher viscosity and, thus, a greater hys-
teresis area (p < 0.05), was reported by Hernández (1996), as
shown in Fig. 2a–d. This demonstrates, according to Holdsworth
(1993), a greater correlation with the thixotropy. Moreover, Her-
nández (1996) states that a thixotropic fluid of higher viscosity
must have a greater hysteresis area than a fluid with lower viscos-
ity, even when an increase occurs in the breaking of the structure
of the less viscous product. The increase in thixotropy as a conse-
quence of higher viscosity was also observed by Tárrega and Cos-
tell (2006) for milk desserts with 6% of inulin and by Domagala
(2008) for yogurts produced with milks form different species.
According to Steffe (1996) and Oliveira et al. (2002) the thixotropy,
a phenomenon commonly detected in fragile agglomerated parti-
cles, such as those of fermented milks, occurs when these are sub-
mitted to a shear force. In this case, the tridimensional structure,
initially formed in the fermentation process, is lost. However, it
can be practically regained, after a period of rest.
The greater hysteresis observed for the fermented milk with
inulin addition may also be related to the amount of total solids,
as shown in Table 1. Mohameed et al. (2004) also associated small
alterations in the concentration of solids to changes in the rheolog-
ical properties of yogurts.
Taking into consideration the storage times and temperatures
evaluated in the rheological study (4 °C and 6 °C) (Fig. 2a–d), it
can be verified that the use of energy is required in order to destroy
the product structure, as defined by González-Tomás et al. (2008)
for products with greater hysteresis, such as prebiotic fermented
milk. During 21 days of storage, the temperature had a significant
effect on the hysteresis of the fermented milks, observing that the
change of 2 °C (from 4 °C to 6 °C) carried out during the rheological
J. Debon et al. / Journal of Food Engineering 99 (2010) 128–135 133
1000 962.9a
919.8a
a
855.3a 852.4a
307.1 823.8b
301.3b 790.0b
Hysteresis loop area (unit)
283.2b 800 746.1b
647.1b
600
400
200
0
1 2 3 4
Storage(days)
Fig. 4. Hysteresis area for prebiotic fermented milk, at temperatures of 4.0 ± 0.1 °C
650 (h) and 6.0 ± 0.1 °C ( ) on storage days 1, 7, 14 and 21.
evaluation, decreased the hysteresis (Figs. 3 and 4). Thus, the de-
crease in the hysteresis (p < 0.05) of the fermented milks with
the increase in temperature could be related to the decrease in
viscosity.
4. Conclusions
The microfiltrate showed low counts for mesophilic and psy-
chrophilic bacteria, being microbiologically appropriate for the
production of prebiotic fermented milk. It was verified that the
addition of inulin increased the total solids content which may
have led to an increase in the acidity of the prebiotic fermented
milk.
In the rheological evaluation carried out for 21 days of product
storage, it was observed that for an increase in the temperature
from 4 °C to 6 °C, the Power Law and Mizrahi and Berk models
were applied successfully to describe the behavior of the flow of
the prebiotic fermented microfiltered milk (with inulin), which
showed a non-Newtonian fluid behavior and the characteristics
of a shear thinning fluid. Furthermore, the addition of inulin to
the fermented milk contributed to an increase in the apparent vis-
cosity when compared with the product without inulin. However,
for the fermented milks the storage time resulted in an increase in
viscosity, while for the variation in temperature investigated the
viscosity decreased. Thixotropic behavior was verified for the fer-
mented milks, however, although the addition of inulin increased
the hysteresis, this was lower with the increase in temperature.
Acknowledgements
The authors express their sincere thanks to the Federal Univer-
sity of Santa Catarina, and also to Beneo™ Orafti and to Victória
Indústria e Comércio de Alimentos Ltda for providing the inulin
and milk samples, respectively.
References
Abu-Jdayil, B., Mohameed, H., 2002. Experimental and modeling studies of the flow
properties of concentrated yogurt as affected by the storage time. Journal of
Food Engineering 52, 359–365.
Adams, D.M., Barach, J.T., Speck, M.L., 1975. Heat resistent proteases produced in
milk by psycrotrophic bactéria of dairy origin. Journal of Dairy Science 58 (6),
828–834.
Akalin, A.S., Tokusoglu, Ö., Gönç, S., Aycan, S., 2007. Occurrence of conjugated
linoleic acid in probiotic yoghurts supplemented with fructooligosaccharide.
International Dairy Journal 17, 1089–1095.
134 J. Debon et al. / Journal of Food Engineering 99 (2010) 128–135
Akin, M.B., Akin, M.S., Kirmaci, Z., 2007. Effects of inulin and sugar levels on the
viability of yogurt and probiotic bacteria and the physical and sensory
characteristics an probiotic ice-cream. Food Chemistry 104, 93–99.
Almeida, K.E., Bonassi, A., Roça, R.O., 2001. Características Físicas e Químicas de
Bebidas Lácteas Fermentadas e Preparadas com Soro de Queijo Minas Frescal.
Ciência e Tecnologia de Alimentos 21 (2), 187–192.
Alparslan, M., Hayta, M., 2002. Rheological and sensory properties of pekmez (grape
molasses)/tahin (sesame paste) blends. Journal of Food Engineering 54 (1), 89–
93.
AOAC, 2005. Official Methods of Analysis, 18th ed. Association Official Analytical
Chemists, Maryland, DC.
APHA, 2001. Compendium of Methods of the Microbiological Examination of Foods,
fourth ed. American Public Health Association, Washington, DC.
Bortolozo, E.Q., Quadros, M.H.R., 2007. Aplicação de inulina e sucralose em iogurte.
Revista Brasileira de Tecnologia Agroindustrial 1 (1), 37–47.
Bourne, M.C., 2002. Food Texture and Viscosity: Concept and Measurement, second
ed. Academic Press, San Diego. 427p.
Brink, J., Langton, M., Stding, M., Hermansson, A., 2007. Simultaneous analysis of the
structural and mechanical changes during large deformation of whey protein
isolate/gelatin gels at the macro and micro levels. Food Hydrocolloids 21 (3),
409–419.
Castro, A.G., 2003. A química e a reologia no processamento de alimentos. Ciência e
Técnica. Instituto Piaget, Lisboa. pp. 37–61.
Castro, F.P., Cunha, T.M., Barreto, P.L.M., Amboni, R.D.M.C., Prudêncio, E.S., 2008.
Effect of oligofructose incorporation on the properties of fermented probiotic
lactic beverages. International Journal of Dairy Technology 62, 74–82.
Coutinho, C.M., Chiu, M.C., Basso, R.C., Ribeiro, A.P.B., Gonçalves, L.A.G., Viotto, L.A.,
2009. State of art of the application of membrane technology to vegetable oils: a
review. Food Research International 42, 536–550.
Cruz, S.A., Farah, M., Zanin, M., Bretãs, R.E.S., 2008. Avaliação das Propriedades
Reológicas de Blendas de PEAD Virgem/PEAD Reciclado. Polímeros: Ciência e
Tecnologia 18 (2), 144–151.
Cueva, O., Kayanush, J., Aryana, K.J., 2008. Quality attributes of a heart healthy
yogurt. LWT – Food Science and Technology 41, 537–544.
Cunha, T.M., Castro, F.P., Barreto, P.L.M., Benedet, H.D., Prudêncio, E.S., 2008.
Avaliação físico-química, microbiológica e reológica de bebida láctea
fermentada e leite fermentado adicionados de probióticos. Semina: Ciências
agrárias 29 (1), 103–116.
Cunha-Neto, O.C., Oliveira, C.A.F., Hotta, R.M., Sobral, P.J.A., 2005. Avaliação físico-
química e sensorial do iogurte natural produzido com leite de búfala contendo
diferentes níveis de gordura. Ciência e Tecnologia de Alimentos 25 (3), 448–453.
Domagala, J., 2008. Sensory evaluation and rheological properties of yoghurts
prepared from goat, cow and sheep milk. Food Science and Technology 11 (3).
Donkor, O.N., Nilmini, S.L.I., Stolic, P., Vasiljevic, T., Shah, N.P., 2007. Survival and
activity of selected probiotic organisms in set-type yoghurt during cold storage.
International Dairy Journal 17, 657–665.
Figueiredo, M.G., Porto, E., 2002. Avaliação do Impacto da Qualidade da Matéria-
Prima no Processamento Industrial do Iogurte Natural. Indústria de Latcicínios 7
(41), 76–80.
Flamm, G., Glinsmann, W., Kritchvsky, D., Prosky, L., Roberfroid, M., 2001. Inulin and
oligofructose as dietary fiber: a review of the evidence. Critical Reviews in Food
Science and Nutrition 41, 353–362.
Fuchs, R.H.B., Tanamati, A.A.C., Antonioli, C.M., Gasparello, E.A., Doneda, I., 2006.
Utilização de Lactobacillus casei e cultura iniciadora na obtenção de iogurte
suplementado com inulina e oligofructose. Boletim CEPPA 24 (1), 83–98.
Gallardo-Escamilla, F.J., Kelly, A.L., Delahunty, C.M., 2007. Mouthfeel and flavour of
fermented whey with added hydrocolloids. International Dairy Journal 17, 308–
315.
Gibson, G.R., Roberfroid, M.B., 1995. Dietary modulation of the human colonic
microbiota: introducing the concept of prebiotics. The Journal of Nutrition 125
(6), 1401–1412.
Gibson, G.R., Probert, H.M., Van Loo, J., Rastall, R.A., Roberfroid, M.B., 2004. Dietary
modulation of the human colonic microbiota: updating the concept of
prebiotics. Nutrition Research Review 17, 259–275.
González-Tomás, L., Bayarri, S., Taylor, A.J., Costell, E., 2008. Rheology, flavour
release and perception of low-fat dairy desserts. International Dairy Journal 18,
858–866.
Gonzalez-Tomás, L., Coll-Marqués, J., Costell, E., 2008. Viscoelasticity of inulin-
starch-based dairy systems. Influence of inulin average chain length. Food
Hydrocolloids 22, 1372–1380.
Guggisberg, D., Cuthbert-Steven, J., Piccinali, P., Bütikofer, U., Eberhard, P., 2009.
Rheological, microstructural and sensory characterization of low-fat and whole
milk set yoghurt as influenced by inulin addition. International Dairy Journal
19, 107–115.
Guven, M., Yasar, K., Karaca, O.B., Hayaloglu, A.A., 2005. The effect of inulin as a fat
replacer on the quality of set-type low-fat yogurt manufacture. International
Journal of Dairy Technology 58, 180–184.
Hassan, B.H., Hobani, A.I., 1998. Flow properties of roselle (Hibiscus sabdariffa L.)
extract. Journal of Food Engineering 35 (4), 459–470.
Hauly, M.C.O., Moscatto, J.A., 2002. Inulina e Oligofrutose: uma revisão sobre
propriedades funcionais, efeito prebiotic e importância na indústria de
alimentos. Revista Semina: Ciências Exatas e Tecnologia de Alimentos 23 (1),
23–26.
Hernández, M.J., 1996. Caracterización reológica de hidrogeles de MCC-
NaCMC + almidón. Tixotropía y sinergismo. Ph.D. Thesis. Universitat de
València, Valência, Spain.
Holdsworth, S.D., 1993. Rheological models used for the prediction of flow
properties of food products. Transactions in Chemical Engineering 71, 139–179.
IAL, 2005. Normas Analíticas do Instituto Adolfo Lutz, 4° ed. Instituto Adolfo Lutz,
São Paulo, DC.
Jelen, P., Lutz, S., 1998. Functional milk and dairy products. In: Mazza, G. (Ed.),
Functional Foods: Biochemical and Processing Aspects. Technomic Publishing,
Lancaster, pp. 357–381.
Karazhiyan, H., Razavi, S.M.A., Phillips, G.O., Fang, Y., Al-Assaf, S., Nishinari, K.,
Farhoosh, R., 2009. Rheological properties of Lepidium Sativum seed extract as a
function of concentration, temperature and time. Food hydrocolloids 23, 2062–
2068.
Kip, P., Meyer, D., Jellema, R.H., 2005. Inulins improve sensoric and textural
properties of low-fat yoghurts. International Dairy Journal 16 (9), 1098–1103.
Lawrence, N.D., Kentish, S.E., O’Connor, A.J., Barber, A.R., Stevens, G.W., 2008.
Microfiltration of skim milk using polymeric membranes for casein concentrate
manufacture. Separation and Purification Technology 60, 127–244.
Lucey, J.A., 2002. Formation and physical properties of milk protein gels. Journal of
Dairy Science 85 (2), 281–294.
Machado, J.C.V., 2002. Reologia e escoamento de fluidos: ênfase na indústria de
petróleo. Editora Interferência Ltda, Rio de Janeiro. pp. 3–109.
Martínez-Villaluenga, C., Frías, J., Gómez, R., Vidal-Valverde, C., 2005. Raffinose
family oligosaccharides from lupin seeds as prebiotics. Application in dairy
products. Journal of Food Protection 68, 1246–1252.
Maskan, M., Gogus, F., 2000. Effect of sugar on the rheological properties of
sunflower oilwater emulsion. Journal of Food Engineering 43, 173–177.
Mistry, V.V., Moubois, J.L., 2004. Application of membrane separation technology to
cheese production. In: Fox, P.F., McSweeney, P.H.L., Cogan, T.M., Guinee, P.T.
(Eds.), Cheese: Chemistry, Physics and Microbiology, third ed. Chapman and
Hall Ltd., pp. 261–284.
Mohameed, H.A., Abu-Jdayil, B., Al-Shawabkeh, A., 2004. Effect of solids
concentration on the rheology of labneh (concentrated yogurt) produced from
sheep milk. Journal of Food Engineering 61, 347–352.
Muir, D.D., 1996. The shelf life of dairy products: factors influencing raw milk and
fresh products. Journal of the Society of Dairy Technology 49 (1), 24–32.
Niness, K.R., 1999. Inulin and oligofructose: what are they? Journal of Nutrition 129
(Suppl.), 1402–1406.
Oliveira, M.N., Sodini, I., Remeuf, F., Tissier, J.P., Corrieu, G., 2002. Manufacture of
fermented lactic beverages containing probiotic cultures. Journal of Food
Science 67, 2336–2341.
Ozer, B.H., Robinson, R.K., Grandison, A.S., Bell, A.E., 1998. Gelation properties of
Milk concentrated by different techniques. International Dairy Journal 8, 793–
799.
Pietrowski, G.A.M., Ott, A.P., Siqueira, C.R., Silveira, J.F., Bayer, K.H., Carvalho, T.,
2008. Avaliação da qualidade microbiológica de leite pasteurizado tipo C
comercializado na cidade de Ponta Grossa-PR. In: VI Semana de Tecnologia em
Alimentos 2 (36), Ponta Grossa/PR.
Roberfroid, M.B., 1999. Concept in functional foods: the case of inulin and
oligofructose. Journal of Nutrition 129 (Suppl.), 1398–1401.
Roberfroid, M.B., 2005. Introducing inulin-type fructans. British Journal of Nutrition
93 (suppl. 1), 13–25.
Ross-Murphy, S.B., 1990. In: Dickinson, E. (Ed.), Food Polymer, Gels and Colloids.
University of Leeds, United Kingdom, pp. 357–368.
Saboya, L.V., Maubois, J.L., 2000. Current developments of microfiltration
technology in the dairy industry. Lait 80, 541–553.
Schaller-Povolny, L.A., Smith, D.E., 2001. Viscosity and freezing point of a reduced
fat ice cream mix as related to inulin content. Milchwissenschaft–Milk Science
International 56, 25–29.
Schröder, M.J.A., Bland, M.A., 1983. Post-pasteurization contamination and shelf life
of HTST – pasteurized milk when filled in a Liqui-PakÒ conventional or Model
820A cartoning machine. Journal of the Society of Dairy Technology 16 (2), 43–
49.
Silva, R.F., 1996. Use of inulin as a natural texture modifier. Cereal Foods World 41
(10), 792–794.
Smithwell, N., Kailasapathy, K., 1995. Psychrotrophic bacteria in pasteurized milk:
problems with shelf life. Journal of Food Technology 50, 28–31.
Soukoulis, C., Lebesi, D., Tzia, C., 2009. Enrichment of ice cream with dietary fibre:
effects on rheological properties, ice crystallisation and glass transition
phenomena. Food Chemistry 115, 665–671.
Statsoft Inc., 2001. STATISTICA (Data Analysis System) Version 6.0. StatSoft Inc.,
Tulsa, OK
Steffe, J.F., 1996. Rheological Methods in Food Process Engineering, second ed.
Freeman Press, East Lansing, 412p.
Tabilo-Munizaga, G., Barbosa-Cánovas, G.V., 2005. Rheology for the food industry.
Journal of Food Engineering 67, 147–156.
Tamime, A.Y., Robinson, R.K., 2007. Yoghurt Science and Technology, third ed.
Woodhead Publishing Limited, Cambridge, 791p
Tárrega, A., Duran, L., Costell, E., 2004. Flow behavior of semi-solid dairy desserts.
Effect of temperature. International Dairy Journal 14, 345–353.
Tárrega, A., Costell, E., 2006. Effect of inulin addition on rheological and sensory
properties of fat-free starch-based dairy desserts. International Dairy Journal
16, 1104–1112.
Teles, C.D., Flôres, S.H., 2007. Influência da adição de espessantes e leite em pó nas
características reológicas do iogurte desnatado. Boletim CEPPA 25 (2), 247–256.
Toneli, J.T.C.L., Mürr, F.E.X., Park, K.L., 2005. Revisão: Estudo da reologia de
polissacarídeos utilizados na indústria de alimentos. Revista Brasileira de
Produtos Agroindustriais 7 (2), 181–204.
 J. Debon et al. / Journal of Food Engineering 99 (2010) 128–135
Toneli, J.T.C.L., Mürr, F.E.X., Martinelli, P., Fabbro, I.M.D., Park, K.J., 2007.
Optimization of a physical concentration process for inulin. Journal of Food
Engineering 80, 832–838.
Tungland, B.C., Meyer, D., 2006. Nondigestible oligo- and polysaccharides (dietary
fiber): their physiology and role in human health and food. Food Science and
Food Safety 1, 73–92.
Tziboula, A., Steele, W., West, I., Muir, D.D., 1998. Microfiltration of milk with
ceramic membranes: influence on casein composition and heat stability.
Milchwissenschaft 53, 8–11.
Villegas, B., Costell, E., 2007. Flow behavior of inulin-milk beverages. Influence of
inulin average chain length and of milk fat content. International Dairy Journal
17, 776–781.
135
Villegas, B., Tárrega, A., Carbonell, I., Costell, E., 2009. Optimising acceptability of new
prebiotic low-fat milk beverages. Food Quality and Preference 21, 234–242.
Ziemer, C.J., Gibson, G.R., 1998. An overview of probiotics, prebiotics and synbiotcs
in the functional food concept: perspectives and future strategies. International
Dairy Journal 8, 473–479.
Zocche, F., Bersot, L.S., Barcellos, V.C., Paranhos, J.K., Rosa, S.T.M., Raymundo, N.K.,
2002. Qualidade microbiológica e físico-química do leite pasteurizado
produzido na região oeste do Paraná. Archives of Veterinary Science 7 (2),
59–67.
Wolfschoon-Pombo, A.F., Granzinolli, G.G.M., Fernandes, R.M., 1983. Sólidos totais
do leite, acidez, pH e viscosidade do iogurte. Revista do Instituto de Laticínios
Cândido Tostes 38, 19–24.