352 International Journal of Food Science and Technology 2007, 42, 352–358

Original article
Effect of fat and non-fat dry matter of milk, and starter type, on the
rheological properties of set during the coagulation process

Özlem Gün & Nursel Develi Işıklı*

Department of Food Engineering, University of Mersin, 33342 Çiftlikköy, Mersin, Turkey
(Received 7 January 2005; Accepted in revised form 5 January 2006)

Summary The effect of fat and non-fat dry matter (NFDM) of milk, and starter type, on the rheological properties of
set yoghurt during the coagulation process was studied using a rotational viscosimeter. The pH was
monitored simultaneously with viscosity readings. Recombined milk was cultured with yoghurt starter
culture (YC), consisting of Streptococcus thermophilus and Lactobacillus bulgaricus, or ABT1, consisting of
S. thermophilus, Lactobacillus acidophilus and bifidobacteria, at 40 C until the pH dropped to 4.7. Yoghurt
made with ABT1 had higher viscosity values than yoghurt made with YC. The NFDM content of milk had a
major effect on the viscosity of curd made using YC or ABT1. For YC and ABT1, three stages were
identified on the viscosity–incubation time curves and viscosity behaviour was different in each stage.
Keywords Fat, non-fat dry matter content, probiotic bacteria, rheology, yoghurt.

solids such as skim milk powder, milk protein concentrate

Introduction
Yoghurt has played an important role in the diet of
people. The essential flora of yoghurt consists of Strep-
tococcus thermophilus and Lactobacillus delbrueckii ssp.
bulgaricus (Tammie, 1985). Over the past decade, the use
of probiotic organisms such as Lactobacillus acidophilus
and bifidobacteria has increased in yoghurt and other
fermented dairy products, mainly because of the healthy
image these dairy products have (Sodini et al., 2002).
Probiotic organisms are defined as living microorganisms
that have beneficial effects on health (Sanders &
Kiaenhammer, 2001). The beneficial effects of probiotics
organisms are attributed to their ability to reduce the
enteric pathogens, supplying enzymes to hydrolyse
lactose, and the ability of some to metabolise dietary
cholesterol and improve intestinal peristaltic activity (Jay,
1992; Salminen & Saxelin, 1996; Saxelin, 1996).
The physical structure of yoghurt is a protein
network. After milk is inoculated with a chosen starter
culture, a progressive lowering of pH occurs, which
leads to changes in the casein micelles associations to
form an aggregated network enclosing fat globules and
serum (Walstra et al., 1999).
Improving the physical and textural properties of
yoghurt has been the subject of investigations for several
authors. Studies have investigated the rheological prop-
erties of yoghurt in relation to the addition of non-fat milk
*Correspondent: Fax: +(0324) 361 00 32;
e-mail: nisikli@yahoo.com
and milk protein powder (Modler et al., 1983; Rhom &
Schmid, 1993), fat (Lucey et al., 1998; De Lorenzi et al.,
1995), polysaccharide content (Sanchez et al., 2000) and
processing parameters (Labropoulos et al., 1983). In
addition, strains of starter cultures used, and incubation
temperature, influence the rheological properties of
coagulum network in yoghurt (Hassan et al., 1996).
The rheological properties of yoghurt are indicative of
gel strength (Hess et al., 1997), and can be used to assess
yoghurt quality (Parnelle-Clunies et al., 1986). Viscosity
development is one important quality control charac-
teristic, particularly for products that we expect to be of
particular consistency in relation to appearance, mouth
feel and further processing (Lewis, 1996). A static
rheological measurement following coagulation usually
necessitates destruction of the sample during the test
(Parnelle-Clunies et al., 1988). Observation of gel
microstructure and dynamic measurement during the
coagulation process are particularly important to under-
stand the changes of structure (Parnelle-Clunies et al.,
1988). To obtain a dynamic measurement of the
coagulation process, various methods including meas-
urements of viscosity, light scattering and laser micros-
copy have been used (Banon & Hardy, 1991; Hassan
et al., 1995; Jumah et al., 2001). Prentice (1992) repor-
ted that viscosimeter is often the preferable for meas-
urement of rheological properties of set and stirred
yoghurt. Monitoring of viscosity during coagulation
process of yoghurt is indicative of changes to yoghurt
texture during manufacture (Shaker et al., 2000).

doi:10.1111/j.1365-2621.2006.01257.x
 2007 The Authors. Journal compilation  2007 Institute of Food Science and Technology Trust Fund

The objectives of this present work are to study the
effect of fat and non-fat dry matter (NFDM) concentra-
tion and starter culture type on the pH and viscosity
behaviour of curd during the coagulation process of milk.
Material and methods
Preparation of yoghurt
Low-heat skim milk powder and water were blended at
room temperature to form an NFDM content of 10%,
12% and 14% w/v in final yoghurt milk.
Reconstituted milk was prepared by dissolving low-
_
heat skim milk powder (Pinar Co., Izmir, Turkey) in
distilled water. The powder contained 3.75% w/w
moisture, 1% w/w fat, 34%w/w protein, 52.5% w/w
lactose and 8.75% w/w ash. The reconstituted skim milk
was stirred for 2 h at room temperature. The reconsti-
tuted milk was divided into three parts. The first part
was used as low-fat milk containing 0.1% w/v milk fat.
The second and third parts were blended with fresh
frozen milk fat (Akdeniz Co., Mersin, Turkey) to bring
the fat contents to 1.6% and 3.2% w/v, respectively.
The recombined milk was heated to 50 C and gently
mixed to disperse the fat. The recombined milk was
homogenised with an Ultra Turrax T25 (IKA Werke,
Janke & Kunkel GmbH & Co. KG, Staufen, Germany)
at 20 000 r.p.m. for 3 min. After heating to 85 C for
10 min, the milk was cooled to 43 C and inoculated
with yoghurt starter cultures (YC) or probiotic cultures
(PC) of yoghurt mix. A mixture of S. thermophilus
(Texel, France) and L. bulgaricus (Rhodia, France) (1:1)
was used as YC and ABT1 (Chr. Hansen A/S, Hoers-
holm, Denmark), consisting of S. thermophilus, L.
acidophilus, bifidobacteria, was used as PC. PC and
YC were in freeze-dried direct-to-vat set form.
Each lot of inoculated milk was distributed in 200 and
500 mL cups. Two hundred millilitre portions of the
inoculated milk were incubated at 43 C until pH 4.7
was reached. Incubation time depended on using starter
culture. After fermentation, the yoghurt samples were
cooled and stored at 4 C for 24 h and then viscosity
was measured.
Five hundred millilitre portions of inoculated milk
were incubated at 43 C in a thermostatically controlled
water bath. Inoculated milk was monitored for pH and
viscosity. Experiments were run in duplicate.
Measurement of viscosity and pH
Viscosity changes during acidification of milk were
monitored using a Brookfield DV-E model digital
viscosimeter (Brookfield Engineering Laboratories
Inc., Middleboro, USA) in 500 mL cups. All viscosity
measurements were conducted with no. 4 spindle at
60 r.p.m. Results were recorded as centipoise units.
 2007 The Authors. Journal compilation  2007 Institute of Food Science and Technology Trust Fund
Effect of fat and NFDM of milk on yoghurt Ö. Gün and N. D. Işıklı 353
pH was measured with glass electrode standardised at
43 C over the pH range 7–4. During the coagulation
process, data obtained from viscosimeter and pH meter
were simultaneously recorded every 10 min.
In yoghurt samples (200 mL) stored at 4 C for 24 h,
viscosity was measured using a Brookfield DV-E model
digital viscosimeter, pH was measured with glass elec-
trode standardised at 4 C over the pH range 7–4.
Statistical analysis
Results were submitted to anova procedures using
Statistica Software (1995). For each main effect, a
multiple comparison of mean was performed using the
least significant difference (l.s.d.) test (P < 0.05).
Results and discussion
The viscosity behaviour during the coagulation pro-
cess of each treatment is presented in Figs. 1 and 2. In
yoghurt produced with PC, viscosity was constant
between 0 and 50 or 80 min, after which there was a
rapid increase in the rate and then a decrease. These
results are comparable with the findings of Shaker
et al. (2000) and Parnelle-Clunies et al. (1988).
Yoghurts produced with YC showed similar trends
to PC yoghurts. Hassan et al. (1995) described struc-
ture formation during the coagulation of milk as a
three-stage process, as lag phase, aggregation and
contraction with rearrangement of casein micelles.
Shaker et al. (2000) reported that viscosity–incubation
time curve can be mathematically separated into
three stages corresponding to different viscosity beha-
viour.
In our study, rate of viscosity change in YC yoghurt
was small compared with change in PC yoghurt. This
observation was confirmed by statistical analysis com-
prising anova (Table 1) and l.s.d. test (data not
shown). The l.s.d. test reported that there was a
significant difference between rate of viscosity change
of PC and YC yoghurt samples after 60 min of
coagulation process. The l.s.d. test also showed that
viscosity values of PC yoghurt were higher than that
of YC yoghurt after 60 min of coagulation process.
However, in PC and YC yoghurt samples containing
0.1% fat, there were no significant differences between
viscosity values after 120 min of coagulation process.
Moreover, viscosity value of PC yoghurt was higher
than that of YC yoghurt after 1-day storage at 4 C
(Table 3). This observation could be related to the
production of exopolysaccharide of ABT1 culture used
in producingPC yoghurt. Use of exopolysaccharide for
producing YCs increases the viscosity and texture of
yoghurt, a fact reported by several researchers (Hassan
et al., 1996; Philippe & Mollet, 2001). As the viscosity
reached the peak point, statistical analysis comprising
International Journal of Food Science and Technology 2007
354 Effect of fat and NFDM of milk on yoghurt Ö. Gün and N. D. Işıklı

1200
10% NFDM

1000 12% NFDM

14% NFDM
800
Viscosity (cp)

600

400

200

–200
20
40
60
80

20
40
60
80

20
40
60
80
100
120
140
160
180

100
120
140
160
180

100
120
140
160
180
Time (min)

Time (min)

Time (min) Figure 1 Effect of NFDM and fat content on

viscosity–incubation time curves in yoghurt
0.1% Fat 1.6% Fat 3.2% Fat produced with PC.

160
10% NFDM
12% NFDM
140
14% NFDM
120
Viscosity (cp)

100

80

60

40

20
20
30
40
60
80

20
30
40
60
80

20
30
40
60
80
100
120
140
160

100
120
140
160

100
120
140
160
Time (min)

Time (min)

Time (min)

Figure 2 Effect of NFDM and fat content on

viscosity–incubation time curves in yoghurt
0.1% Fat 1.6% Fat 3.2% Fat produced with YC.

anova (Table 2) and l.s.d. test reported that an
interaction among the NFDM, fat content and starter
culture type had significant effects on the viscosity at
the peak point (Table 2). The highest rate of viscosity
in each type of yoghurt occurred at 14% w/v NFDM
(Fig. 3). In PC yoghurts, an increase in fat level (of
the samples containing 10% and 14% NFDM)
increased the rate of viscosity. In contrast, YC yoghurt
did not show an increase in the rate of viscosity
(Fig. 3).
The pH value decreased in all samples during the
coagulation (Figs. 4 and 5). Fermentation was stopped
when pH value reached 4.7. Fermentation time of
yoghurt using PC (180 min) was longer than the yoghurt
using YC (150 min). A mixed culture composed of
S. thermophilus ST7 and L. bulgaricus LB12 presented a
high interaction factor and permitted a short fermenta-
tion time (Sodini et al., 2002).
During the coagulation process, apparent viscosity
remains constant before pH drops to 5.8 and increases

International Journal of Food Science and Technology 2007  2007 The Authors. Journal compilation  2007 Institute of Food Science and Technology Trust Fund
 Effect of fat and NFDM of milk on yoghurt Ö. Gün and N. D. Işıklı 355

Table 1 Analysis of variance for the response

of dependent variables Viscosity (cP) pH

Source of variation df Mean square Mean square

Starter culture 1 831553.1*** 0.817***

NFDM 2 76287.6*** 0.094***
Fermentation time 13 82781.7*** 7.424***
Fat 2 8944.6*** 0.225***
Starter culture · NFDM 2 29296.8*** 0.017***
Starter culture · fermentation time 13 51148.2*** 0.152***
NFDM · fermentation time 26 17932.8*** 0.026***
Starter culture · fat 2 45476.3*** 0.923***
NFDM · fat 4 3462.8*** 0.076***
Fermentation time · fat 26 20363.2*** 0.023***
Starter culture · NFDM · fermentation time 26 22216.2*** 0.005***
Starter culture · NFDM · fat 4 8895.3*** 0.104***
Starter culture · fermentation time · fat 26 16287.4*** 0.008***
NFDM · fermentation time · fat 52 14679.6*** 0.013***
Starter culture · NFDM · fermentation time · fat 52 12574.7*** 0.012***
Error 252 235.3 0.0002

df ¼ degree of freedom, NFDM ¼ non-fat dry matter.

*** P < 0.001.

Table 2 Analysis of variance for the response of dependent variables at (Prentice, 1992). In our study, viscosity of yoghurt
the peak point of viscosity produced with PC increased dramatically as pH
dropped to 5.1 (or 5.2). When YC was used, viscosity
Viscosity (cP) pH generally increased, as pH dropped to 5 or 5.1. These
Source of variation df Mean square Mean square
results were similar to the finding of Shaker et al. (2000),
and may be concerned with the effect of pH on
Starter culture 1 1690000.0*** 0.213*** microstructure during the coagulation process in milk.
NFDM 2 176287.4*** 0.009*** Previous research by Hassan et al. (1995) reported that
Fat 2 14215.9*** 0.006*** as the pH drops from 6.4 to 5.5, the size and distribution
Starter culture · NFDM 2 82456.3*** 0.014*** of casein micelles is uniform, whereas when the pH
Starter culture · fat 2 55570.6*** 0.025***
drops to 5.3 the casein appear as large micelles, and
NFDM · fat 4 3374.5*** 0.002**
Starter culture · NFDM · Fat 4 14959.0*** 0.001*
large casein aggregates initiate formation of network. In
Error 18 11.5556 0.0004
our study, increase in viscosity at pH 5.2 indicated
coagulation of casein, and the decrease in viscosity at
df ¼ degree of freedom, NFDM ¼ non-fat dry matter. pH range 5.1–4.7 might be explained with rearrange-
*P < 0.05, **P < 0.01, ***P < 0.001. ment of casein aggregated. Parnelle-Clunies et al. (1988)
noted that dissociation of the casein micelles appeared at
pH 5.1 and initial changes in the rate of viscosity
Table 3 Viscosity value of yoghurt samples after 1-day storage at 4 C occurred at the point of micellar dissociation. They also
reported that the level of ionic Ca was close to
Viscosity value (cP)
maximum when micellar dissociation occurred.
10% NFDM 12% NFDM 14% NFDM As the viscosity reached the peak point, an interac-
tion among the NFDM and fat content and starter
Fat (%) Fat (%) Fat (%)
culture type had significant effect on pH (Table 2).
Starter culture 0.1 1.6 3.2 0.1 1.6 3.2 0.1 1.6 3.2 Fig. 6 illustrates that the highest pH values were
displayed using PC, at the peak point of viscosity.
Probiotic culture 1391 1470 1374 2005 2128 2108 2378 2600 2657 This result indicated that starter culture type has a
Yoghurt culture 1040 785 312 1418 1360 1070 1459 1585 1490 significant effect on yoghurt viscosity and pH at the
peak point of viscosity. The statistical analysis reported
that within PC yoghurt, the highest pH value was
after pH reached about 5.0 (viscosity reached to a peak obtained for the 0.1% w/v fat content. At 0.1% fat
point) (Prentice, 1992). At this point, fat globules and content, there was no significant difference between pH
serum are considered to be entrapped in a network values of PC yoghurt containing 10%, 12% and 14%

 2007 The Authors. Journal compilation  2007 Institute of Food Science and Technology Trust Fund International Journal of Food Science and Technology 2007
356 Effect of fat and NFDM of milk on yoghurt Ö. Gün and N. D. Işıklı

1200
YC
1000 h
PC
g
800
Viscosity (cp)

600 f f
f e f
e
400 d

200 c abc
ab a ab bc
c ab bc
0

–200 Figure 3 Effect of fat · NFDM · starter cul-

10 12 14 10 12 14 10 12 14 ture interaction on viscosity at peak point of
0.1% Fat 1.6% Fat 3.2% Fat viscosity. Fat and NFDM with no common
letter differ according to the l.s.d. test
NFDM (%) (P £ 0.05).

7.0
10% NFDM
6.5 12% NFDM
14% NFDM

6.0
pH

5.5

5.0

4.5

4.0
Time (min)
20
40
60
80
100
120
140
160
180
Time (min)
20
40
60
80
100
120
140
160
180
Time (min)
20
40
60
80
100
120
140
160
180

Figure 4 Effect of NFDM and fat content

on pH–incubation time curves in yoghurt
0.1% Fat 1.6% Fat 3.2% Fat produced with PC.
6.4
10% NFDM
6.2 12% NFDM
6.0 14% NFDM

5.8
5.6
pH

5.4
5.2
5.0
4.8
4.6
4.4
20
30
40
60
80
100
120
140
160
20
30
40
60
80
100
120
140
160

20
30
40
60
80
100
120
140
160
Time (min)

Time (min)

Time (min)

Figure 5 Effect of NFDM and fat content on

pH–incubation time curves in yoghurt pro-
0.1% Fat 1.6% Fat 3.2% Fat duced with YC.

International Journal of Food Science and Technology 2007  2007 The Authors. Journal compilation  2007 Institute of Food Science and Technology Trust Fund

5.25
i i
5.20
5.15
5.10
5.05
pH
5.00
4.95
b YC
4.90
a
4.85
4.80
Figure 6 Effect of fat · NFDM · starter cul-
10 12
ture interaction on pH at peak point of visc-
0.1% Fat
osity. Fat and NFDM with no common letter
differ according to the l.s.d. test (P £ 0.05).
w/v NFDM; an increase in fat led to lower pH. This
effect of fat on PC action remains unclear. In YC
yoghurt, an increase in NFDM and fat content led to
higher pH at a peak point of viscosity. The effect of
NFDM on pH was similar to the effect on viscosity.
Within the PC and YC yoghurts, as an NFDM level
was increased from 10% w/v to 14% w/v, generally a
peak viscosity at 14% w/v NFDM was higher than that
at 10% and 12% w/v NFDM while fermentation time
to reach a peak viscosity was shortened. It is known
that the casein content was one of the important
factors in the curd formation and curd structure.
Increase in NFDM content in milk increased the casein
content of yoghurt milk and this attributed to an
increased water-binding capacity in the milk proteins
(Walstra et al., 1999). Therefore, the higher the NFDM
used, the higher viscosity values were obtained.
Conclusion
This study shows that the behaviour of PC and YC is
different, and is related to milk composition. Within
each type of culture, the viscosity of yoghurt milk during
the coagulation process was affected by the milk
content, and was higher for milk with higher NFDM
content. An increase in fat level improved the viscosity
for PC. However, a poor viscosity has been observed as
a result of the high level of fat added for YC. Type of
starter culture affected the rheology nature of yoghurt
during the coagulation process.
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