LWT - Food Science and Technology 90 (2018) 620–626

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LWT - Food Science and Technology

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Fortification of set-type yoghurts with Elaeagnus angustifolia L. flours: Effects T

on physicochemical, textural, and microstructural characteristics
Hale İnci Öztürka,∗, Sümeyye Aydına, Didem Sözerib, Talha Demircia, Durmuş Sertc, Nihat Akına
a
Department of Food Engineering, University of Selcuk, Konya 42050, Turkey
b
Department of Food Engineering, University of Namık Kemal, Tekirdag 50860, Turkey
c
Department of Food Engineering, University of Necmettin Erbakan, Konya 42060, Turkey

A R T I C L E I N F O A B S T R A C T

Keywords: The effect of peeled oleaster (Elaeagnus angustifolia L.) flour (PO) and unpeeled oleaster flour (UPO) in different
Yoghurt levels (1% and 2%) on the quality parameters of set type yoghurt was investigated throughout 28 days of
Fortification storage. Acidification kinetics, water holding capacity, syneresis level, textural characteristics and micro-
Acidification structure were firstly measured. Additionally, influence of oleaster flours on DPPH and ABTS+ radical scaven-
Texture
ging activity and total phenolic content of yoghurt samples was also assessed. Besides, all the products were
Microstructure
subjected to sensorial preference test. Enrichment with 2% PO and UPO reduced fermentation time by 31 and
37 min, respectively. Addition of PO and UPO increased cohesiveness and the viscosity index, furthermore,
syneresis decreased. Oleaster supplementation to yoghurt significantly increased scavenging activities of DPPH
and ABTS+ radicals and the highest activities were determined in yoghurts fortified with 2% UPO. The results
demonstrated that yoghurt with reduced syneresis, improved functional properties, and enhanced some textural
characteristics can be achieved by a 2% UPO addition. Also, surprisingly yoghurts containing 2% UPO had
similar consumer overall preference scores comparing to plain yoghurt.

1. Introduction
Yoghurt is a fermented dairy product made by the activity of
Streptococcus thermophilus (S. thermophilus) and Lactobacillus delbrueckii
ssp. bulgaricus (L. bulgaricus), and it is widely consumed around the
world (Akın, 2006). Textural properties of yoghurt are very important
for consumer acceptability, product development, and quality assess-
ment (Benezech & Maingonnat, 1994). Yoghurt gel formation is caused
by the aggregation of casein micelles after decreased pH in the milk due
to lactic acid production. The three-dimensional protein network is
directed by the total solids content, acidity production rate, and pro-
teolytic activity of the starter bacteria (Lee & Lucey, 2004). Yoghurt gel
structure is principally associated with the shape and strength of the
protein network structure. Increasing the interconnections between
milk proteins by reducing syneresis is an important approach in the
development of the textural quality of any yoghurt (Shi, Han, & Zhao,
2016).
Some studies have been performed to enhance yoghurt quality such
as reducing syneresis, enhancing viscosity and textural characteristics
by adding whey proteins (Laiho, Williams, Poelman, Appelqvist, &
Logan, 2017), fiber-rich fruit peel powder (do Espírito Santo, Perego,
Converti, & Oliveira, 2012), seed mucilage or guar gum (Mudgil, Barak,
& Khatkar, 2016), exopolysaccharides (Zhang, Folkenberg, Qvist, &
Ipsen, 2015), and hydrocolloids such as gelatin (Shi et al., 2016).
Elaeagnus angustifolia L. which is called oleaster or Russian olive
pertains to the genus Elaeagnus of Elaeagnaceae family. E. angustifolia L.
has small reddish-brown elliptic fruits and grows in a wide geographical
area including Asia, Europe and North America (Fonia, White, & White,
2009). Oleaster fruits have high nutritional value due to the comprising
of proteins, carbohydrates, vitamins, minerals, phenolic compounds,
antioxidants, and fibers (Abizov, Tolkachev, Mal'tsev, & Abizova,
2008). Oleaster fruit consists of 27.1% glucose, 22.3% fructose, 12%
protein, 4.65% ascorbic acid and 1% ash (Akbolat, Ertekin, Menges,
Guzel, & Ekinci, 2008; Ayaz & Bertoft, 2001). Also, phytochemical re-
searches have shown that oleaster fruit is rich in flavonoids, phenolic
acids and vitamins (Abizov et al., 2008; Ayaz & Bertoft, 2001), thus,
oleaster fruits showed high antioxidant activity (Faramarz, Dehghan, &
Jahanban-Esfahlan, 2015). Çakmakçı et al. (2015) determined an in-
crease in total phenolic content and antioxidant activity of ice cream
fortified with oleaster flour. Besides, Sahan et al. (2015) noted that
oleaster flour with high fiber content (20–30%) showed high water
absorption potential and an enhancing effect on emulsion stability.

∗
Corresponding author. Department of Food Engineering, Faculty of Agriculture, University of Selcuk, Konya 42050, Turkey.
E-mail address: haleinciozturk@selcuk.edu.tr (H.İ. Öztürk).

https://doi.org/10.1016/j.lwt.2018.01.012
Received 29 October 2017; Received in revised form 13 December 2017; Accepted 4 January 2018
Available online 08 January 2018
0023-6438/ © 2018 Elsevier Ltd. All rights reserved.
H.İ. Öztürk et al.
Although there were some previous studies regarding many new
ingredients to develop yoghurt quality parameters and add beneficial
properties, we met no studies about the incorporation of oleaster as an
ingredient in the yoghurt formulation. On the basis of this background,
the present research was undertaken to examine the influence of
oleaster flour supplementation on the textural, microstructural, physi-
cochemical and sensory attributes of yoghurt. Moreover, the effects of
oleaster fortification on acidification kinetics, total phenolic contents
and antioxidant activity were evaluated.
2. Materials and methods
2.1. Experimental materials
Raw cow milk was collected from Selcuk University Dairy Farm,
Konya, Turkey and medium-heat skim milk powder (34.5% protein,
3.5% moisture, 7.2% ash, 55% lactose, pH:6.55 and 0.112% titratable
acidity) was provided from ENKA Dairy Product, Konya, Turkey.
Commercial freeze-dried yoghurt starter culture YF-L901 consisting of
S. thermophilus and L. bulgaricus supplied from Chr's Hansen-Peyma
(Istanbul) was used in yoghurt production.
Two different supplements, including PO and UPO, were prepared.
In the production of the PO, peeled and seedless oleaster fruits were
kept in hot air oven at 30 °C over the night and then were ground in a
hammer mill (Falling Number-3100 Laboratory Mill, Huddinge,
Sweden). The same operations except peeling process were also carried
out to obtain UPO.
2.2. Set-type yoghurt processing and sample treatments
Dry matter of milk was adjusted to 16% by adding medium-heat
skim milk powder and it was then divided into 5 experimental groups.
PO and UPO were added separately at 1% and 2% ratios to experi-
mental groups except control sample. Each experimental mix was pas-
teurized at 90 °C for 10 min and then rapidly cooled to 42 °C. The yo-
ghurt mixes were inoculated with 2% starter culture and they were then
put into 150 mL sterile plastic containers. All containers incubated at
42 °C until the pH reached 4.5 and were stored at 4 °C for 28 days.
Yoghurt samples were identified as follows: with peeled oleaster flour
(YPO) and with unpeeled oleaster flour (YUPO).
2.3. Monitoring of acidification kinetics
The changes in pH values during fermentation were monitored until
the pH 4.5. Four parameters were utilized to characterize the process
kinetics: the maximum acidification rate (Vmax, 10−3 upH min−1), the
time at which the maximum acidification rate (TVmax, h), the time re-
quired to reach pH 5.0 (TpH 5.0, h), and the time required to complete
fermentation (TpH 4.5, h) (Marafon, Sumi, Alcantara, Tamime, & De
Oliveira, 2011).
2.4. Assessment of physicochemical characteristics
Total solids, ash, fat and color characteristics of yoghurt samples
were determined according to the methods described by Bradley et al.
(1992) on day 14. Furthermore, titratable acidity, pH (Bradley et al.,
1992), water holding capacity and syneresis (Isanga & Zhang, 2009)
were observed at 1, 14, and 28 days of cold storage.
2.5. Extraction for antioxidant and total phenolic content analyses
For extraction, 5 g of yoghurt samples were mixed with 25 mL of
75% methanol solution. Thereafter the mixes were homogenized by
ultra-turrax homogenizer (IKA T-25, Germany) and subjected to cen-
trifugation (Nüve NF 800R, Turkey) at 7200 rpm for 10 min at 4 °C.
Finally, the acquired supernatants were filtered by Whatman No.1 and
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LWT - Food Science and Technology 90 (2018) 620–626
the extracts were stored at 4 °C for analysis.
2.6. Assessment of antioxidant activity with radical scavenging methods
Antioxidant activities of yoghurt samples were determined by ABTS
(2,2′-Azino-bis-3-ethylbenzothiazoline-6-sulfonic) (Re et al., 1999) and
DPPH (1,1-diphenyl-2-picrylhydrazyl) (Brand-Williams, Cuvelier, &
Berset, 1995) radical scavenging methods. ABTS radical scavenging
activity was detected at 734 nm and the measured absorbances were
converted to Trolox equivalent antioxidant capacity (TEAC). The results
of ABTS radical scavenging activity were expressed as μM Trolox
equivalent per gram of yoghurt samples. DPPH radical scavenging ac-
tivity was examined at 517 nm and the results were given as % in-
hibition.
2.7. Measurement of total phenolic content
The total phenolic content (TPC) was determined by using the
method of Tseng and Zhao (2013). Reaction absorbance was measured
at 765 nm. The results, that were calculated based on the gallic acid
curve, were expressed as microgram gallic acid equivalent (GAE) per
gram of yoghurt samples.
2.8. Texture profile analysis and viscosity index
Firmness, consistency, cohesiveness and viscosity index values were
obtained by TA-XT2 Texture Analyzer (Stable Micro Systems, England)
equipped with a 500 N compression load cell and operating at 1 mm s−1
head speed. The probe was a 25-mm acrylic cylinder moved speed of
5 mm s−1 and test speed of 1 mm s−1 through 10 mm within the
sample. Textural characteristics were examined over cold storage and
the results were given as the mean of three measurements.
2.9. Microstructure analysis
After two weeks of storage, the confocal laser scanning microscopy
(CLSM) analysis was carried out using an inverted confocal laser
scanning microscope (A1/A1R, Nikon, Japan) equipped with Nikon
Plan Fluor, PA:0.30 objective lens. For CLSM observation, yoghurt
samples were stained with Rhodamine B (Fluka, Sigma-Aldrich,
Missouri, USA) in water solution (0.2%, w/v) for 30 min at room tem-
perature. CLSM observation was carried out in a dark and an Ar laser
line (488 nm) was employed as a light source to excite fluorescent dyes
Rhodamine B. For the yoghurt analysis, at least two CLSM images were
taken for each yoghurt treatment in each trial. Two trials of yoghurt
were carried out and hence a minimum of four images were collected
for each treatment.
2.10. Sensory evaluation
Sensory acceptability test of yoghurt samples was assessed by a
trained panel of seven members using five-point hedonic scale (1: un-
acceptable; 5: excellent) (Karagul-Yuceer & Drake, 2006). Panelists
appraised flavor, color and appearance, body and texture, odour, and
general acceptability of yoghurt samples on the 14th day of storage.
2.11. Statistical analysis
The results were analyzed with one-way ANOVA to identify sig-
nificant differences between means of samples and storage days. All
data were expressed as a mean ± standard deviation of the three re-
plicates. The means of results were compared by the Tukey test with a
confidence interval set at 95%.
H.İ. Öztürk et al. LWT - Food Science and Technology 90 (2018) 620–626

Table 1 crust in ice cream samples. When compared to color values of YPO and
Kinetic parameters of acidification of yoghurt samples. YUPO, it was observed that oleaster peel diminished the L* values and
did not change the a* values.
Samples Vmax (10−3 upH TVmax (h) TpH 5.0 (h) TpH 4.5 (h)
min−1) The TPCs of yoghurt samples were determined between 3.25 and
9.76 μg GAE∗ g−1 (Table 2). Addition of oleaster flours increased the
Control 21.31 ± 0.02a 2.52 ± 0.03a 3.15 ± 0.01a 4.40 ± 0.02a TPCs of yoghurt samples. TPC increased significantly by increasing
1% YPO 16.08 ± 0.14d 2.17 ± 0.02b 2.43 ± 0.03b 4.14 ± 0.01b
amount of added PO and UPO (P < .05) and the highest TPC was
2% YPO 15.10 ± 0.17e 1.43 ± 0.03c 2.32 ± 0.03c 4.09 ± 0.04bc
1% YUPO 20.17 ± 0.29b 2.54 ± 0.01a 2.39 ± 0.01b 4.30 ± 0.05a determined in 2% YUPO. Relatively higher phenolic content in 2%
2% YUPO 17.00 ± 0.50c 2.15 ± 0.05b 2.22 ± 0.03d 4.03 ± 0.06c YUPO than those of YPO and control is in agreement with the report of
Çakmakçı et al. (2015) who found that the TPC of sample enriched with
YPO: yoghurt with peeled oleaster flour; YUPO: yoghurt with unpeeled oleaster flour. oleaster crust was higher than the sample containing oleaster flour.
Vmax: maximum acidification rate; Tmax: time to reach Vmax; TpH 5.0: time to obtain pH 5.0;
The scavenging activities of ABTS and DPPH radicals of yoghurt
TpH 4.5: time to obtain pH 4.5.
samples were increased by addition of the PO and UPO (Table 2). The
Results are expressed as the mean ± standard deviation (n = 3).
Different letters in the same column are significantly different (P < .05). increases in antioxidant activity were parallel to the increase in the
added flour rate. Faramarz et al. (2015) reported that oleaster peel
3. Results and discussion showed higher DPPH inhibition than oleaster flesh. In accordance with
this report, the highest DPPH inhibition was observed in 2% YUPO and
3.1. Acidification kinetics during fermentation the addition of 2% UPO increased the DPPH radical scavenging activity
about 5-fold compared to the control yoghurt. Likewise, the highest
The results of the fermentation kinetics of enriched and control ABTS radical scavenging activity was also determined in 2% YUPO.
yoghurts are demonstrated in Table 1. The maximum acidification rates Higher phenolic content in UPO than that of PO due to the presence of
(Vmax) were determined between 15.10 and 21.31 10−3 pH units per polyphenols such as coniferyl alcohol, (E)-isoeugenol in oleaster peel
min. Vmax was significantly reduced by oleaster flour supplementation may have been responsible for higher antioxidant activity of YUPO
(P < .05). Several studies reported that oleaster fruits contained or- (Farzaei, Bahramsoltani, Abbasabadi, & Rahimi, 2015).
ganic acids such as citric acid and malic acid, phenolic compounds and
proteins (Sahan et al., 2015; Çakmakçı et al., 2015). do Espírito Santo 3.3. Post acidification and titratable acidity
et al. (2012) have been reported that these components with buffering
capacity can reduce Vmax. The results of titratable acidity and post acidification during cold
The time to reach maximum acidification rate (TVmax) was de- storage are demonstrated in Table 3. The yoghurt samples, except
creased by the addition of oleaster flours except 1% UPO (P < .05). merely 2% YUPO, had the same behavior during storage regarding the
Furthermore, 2% YPO presented the lowest TVmax of the yoghurt sam- maintenance of the pH until day 14. Likewise, several workers have
ples. The fortification with PO and UPO also reduced the time to reach reported that stable pH trend from 1st day to 14th day of storage in
pH 5.0 (TpH 5.0) and the shortest TpH 5.0 was determined in 2% YUPO. In consistent with our findings (Comunian et al., 2017; Guven, Yasar,
1% YPO, 2% YPO and 2% YUPO, fermentation time (TpH 4.5) was re- Karaca, & Hayaloglu, 2005). In the present study, following 14th day,
duced by 5.91, 7.05 and 8.41%, respectively, compared to plain yo- the pH values of control and UPO supplemented yoghurts decreased till
ghurt. The oleaster flour supplementations, excluding 1% UPO, statis- the end of the storage, whereas pH of yoghurts enriched PO remained
tically accelerated the fermentation (P < .05). TpH 4.5 values of stable. Paseephol and Sherkat (2009) were found same results when
yoghurts enriched with PO and UPO were found to be shorter than that produced yoghurt with Jerusalem artichoke inulin throughout 28 days
of yoghurt added with passion fruit peel powder (do Espírito Santo of storage in parallel with our steady pH trend of YPO. However,
et al., 2012). The decrease in fermentation time may have been caused Comunian et al. (2017) found a decline in pH values after stable trend
by 20.67–30.65% dietary fiber, 27.1% fructose and 22.3% glucose up to 14th day when experienced yoghurt containing free and en-
found in oleaster fruit (Ayaz & Bertoft, 2001; Sahan et al., 2015). Our capsulated echium oil, phytosterol and sinapic acid similar to our re-
results are supported by the report of Sah, Vasiljevic, McKechnie, and sults about UPO supplemented yoghurts. These difference of pH ten-
Donkor (2016) who determined that dietary fiber ensured additional dency between two treatments in the current study may be due to the
carbohydrate sources for yoghurt bacteria. The shortening in fermen- benefit of added UPO and PO for yoghurt starter cultures throughout
tation time can be advantageous to yoghurt manufacturers in terms of the shelf-life. Indeed, the counts of L. bulgaricus had higher (P < .05)
reducing the cost of production. increments in 1% and 2% YPO compared to those in 1% and 2% YUPO
from the first day up to 28th day (data not shown).
The titratable acidity values for all trials exhibited increase with
3.2. Physicochemical attributes, total phenolic content and antioxidative time from the initial day to end day of storage but the rate of increase
activity was markedly higher (P < .05) in the presence of PO comparing to the
other yoghurt samples. A similar behavior of reduction in titratable
Physicochemical characteristics of yoghurt samples are shown in acidity from the first day to 28th day are observed by Agil et al. (2013)
Table 2. The total solids, fat and ash contents of the yoghurt samples who reported sharper increase throughout the shelf-life in yoghurts
ranged from 15.34 to 17.26%, 3.00–3.50% and 1.29–1.39%, respec- containing lentil. The higher titratable acidity values during storage of
tively. Total solids concentration was found as the lowest in plain yo- PO enriched yoghurts can be explained by the greater use of this peeled
ghurt, while it was highest in yoghurts enriched with 2% oleaster oleaster flour by starter cultures compared to other samples. This case
flours. Guggisberg, Cuthbert-Steven, Piccinali, Bütikofer, and Eberhard was not surprising because the percentage of nutrients that can be used
(2009) stated that total solids increased with increasing concentration by the starter cultures is higher in peeled oleaster enrichments than that
of additive in yoghurt formulation. The brightness value (L*) was de- of unpeeled oleaster because of the presence and the inability of the
creased by the addition of 2% PO, 1% UPO and 2% UPO, while the peel to be used by bacteria.
lowest greenness value (a*) was detected in 2% YPO and 2% YUPO.
Moreover, the yellowness value (b*) increased with the addition of 2% 3.4. Water holding capacity and syneresis
PO and 2% UPO. These results are consistent with the findings of
Çakmakçı et al. (2015) who pointed out that greenness values de- No substantial difference was found between the water holding
creased, while yellowness values increased by adding oleaster flour and capacity (WHC) of yoghurt samples on the first day of cold storage and

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H.İ. Öztürk et al. LWT - Food Science and Technology 90 (2018) 620–626

Table 2
Physicochemical characteristics, antioxidant capacity and total phenolic contents of yoghurt samples.a

Control 1% YPO 2% YPO 1% YUPO 2% YUPO

Fat (%) 3.20 ± 0.05bc 3.00 ± 0.07c 3.40 ± 0.28ab 3.20 ± 0.04bc 3.50 ± 0.14a
Ash (%) 1.29 ± 0.00b 1.34 ± 0.01a 1.38 ± 0.01a 1.38 ± 0.03a 1.39 ± 0.02a
Total solids (%) 15.34 ± 0.08c 16.05 ± 0.14b 17.21 ± 0.00a 16.10 ± 0.23b 17.26 ± 0.16a
L* 88.87 ± 1.27a 88.23 ± 0.37ab 87.11 ± 0.27bc 86.84 ± 0.54c 86.49 ± 0.43c
a* −4.10 ± 0.28d −2.85 ± 0.13c −1.88 ± 0.11a −2.30 ± 0.22b −1.76 ± 0.26a
b* 8.12 ± 1.87b 9.64 ± 0.48ab 10.07 ± 0.17a 8.95 ± 0.62ab 10.02 ± 0.41a
DPPH (% Inhibition) 1.22 ± 0.01e 2.31 ± 0.01d 2.93 ± 0.04b 2.56 ± 0.08c 6.09 ± 0.07a
ABTS (μM Trolox g−1) 192.49 ± 1.90d 240.11 ± 3.55c 252.69 ± 0.55b 252.98 ± 5.76b 269.65 ± 7.23a
TPC (μg GAE g−1) 3.25 ± 0.02e 4.15 ± 0.05d 4.95 ± 0.03c 7.45 ± 0.03b 9.76 ± 0.04a

YPO: yoghurt with peeled oleaster flour; YUPO: yoghurt with unpeeled oleaster flour.
TPC: total phenolic content; GAE: gallic acid equivalent.
Results are expressed as the mean ± standard deviation (n = 3).
Different letters in the same row are significantly different (P < .05).
a
The results were given for 14 days.

Table 3 lowest syneresis value was determined in 2% YUPO. The syneresis va-
Changes in post acidification and titratable acidity during cold storage. lues of YPO and YUPO diminished with progress of storage days, while
no change in the syneresis value of the control sample was observed
Samples d1 d14 d28
during the storage. The decrement in syneresis was possibly a con-
pH Control 4.27 ± 0.01a, b
4.27 ± 0.01a, a
4.09 ± 0.01b, b sequence of water binding abilities of PO and UPO in accordance with
1% YPO 4.08 ± 0.00a, e 4.11 ± 0.01a, c 4.10 ± 0.01a, b the findings of Sahan et al. (2015).
2% YPO 4.13 ± 0.00a, d 4.12 ± 0.01a, c 4.13 ± 0.00a, a
1% YUPO 4.25 ± 0.01a, c 4.26 ± 0.02a, a 4.13 ± 0.01b, a
2% YUPO 4.34 ± 0.01a, a 4.21 ± 0.01b, b 4.14 ± 0.01c, a 3.5. Textural characteristics
Titratable Control 1.58 ± 0.02b, a 1.60 ± 0.01b, b 1.73 ± 0.07a, c
acidity 1% YPO 1.58 ± 0.01c, a 1.95 ± 0.01b, a 2.13 ± 0.06a, a
(% lactic 2% YPO 1.60 ± 0.01c, a 1.92 ± 0.01b, a 1.97 ± 0.00a, b The firmness values of yoghurt samples and the changes during cold
acid) 1% YUPO 1.57 ± 0.01b, a 1.57 ± 0.01b, b 1.67 ± 0.00a, c storage are shown in Fig. 1. The highest firmness value was detected in
2% YUPO 1.51 ± 0.00c, b 1.60 ± 0.02b, b 1.69 ± 0.02a, c the control sample (P < .05) on first day of cold storage and a decrease
in firmness was determined depending on the addition of PO and UPO.
YPO: yoghurt with peeled oleaster flour; YUPO: yoghurt with unpeeled oleaster flour.
Damin, Minowa, Alcantara, and Oliveira (2008) notified that long fer-
Results are expressed as the mean ± standard deviation (n = 3).
Different letters in the same row and coloumn are significantly different (P < .05).
mentation time improved the firmness of yoghurt gel, therefore, control
First letters show differences between the storage times and second letters present the sample with longer fermentation time compared to YPO and YUPO
differences between the yoghurt samples. demonstrated greater gel firmness. Besides, Paseephol, Small, and
Sherkat (2008) reported that the fibers were caused to form a soft gel
the WHC values ranged from 58.25 to 59.95% (Table 4). An increase in due to their dispersion in casein micelles and thereby their presence in
WHCs of 2% YPO and 2% YUPO was determined at the end of the the protein matrix. This may explain that the firmness values of yoghurt
storage. This could be associated with dietary fiber content in PO and samples containing PO and UPO were lower than the plain yoghurt. In
UPO. In a previous study, it was reported that dietary fiber absorbed this study, gel firmness of YPO and YUPO increased at the end of the
serum released from yoghurt gel structure (García-Pérez et al., 2005). storage in comparison with the first day, however, gel firmness of
Sahan et al. (2015) informed that oleaster flour presented high water control sample decreased. The enhancement in firmness of YPO and
absorption capacity. YUPO may be related to ascending water absorption ability of oleaster
The addition of PO and UPO significantly affected syneresis flours.
(P < .05). On the first day of storage, the syneresis values of yoghurt The addition of PO and UPO declined the consistency on the first
samples were between 13.57 and 19.72% (Table 4). Whey separation day of the cold storage (Fig. 1). A decrease in consistency was observed
was significantly decreased with PO and UPO fortifications and the in YPO and YUPO with increasing supplement rate (P < .05). This
result accords with the findings of Guven et al. (2005) who reported

Table 4
Water holding capacity and syneresis of yoghurt samples during cold storage.

Samples d1 d14 d28

Syneresis (%) Control 19.72 ± 0.03a, a 18.58 ± 1.19a, a 18.76 ± 0.84a, a

1% YPO 17.92 ± 0.02a, b 13.52 ± 0.56b, b 13.00 ± 0.48b, b
2% YPO 15.40 ± 0.10a, d 15.04 ± 0.04a, ab 11.82 ± 0.94b, bc
1% YUPO 16.84 ± 0.68a, c 16.86 ± 3.66a, ab 9.76 ± 1.56b, c
2% YUPO 13.57 ± 0.27a, e 12.86 ± 1.18a, b 9.56 ± 1.00b, c
Water holding capacity (%) Control 58.25 ± 0.75a, a 60.55 ± 1.15a, c 60.80 ± 1.80a, ab
1% YPO 59.10 ± 4.90a, a 60.20 ± 0.40a, c 59.35 ± 0.05a, b
2% YPO 58.75 ± 0.45b, a 62.25 ± 0.35a, b 62.30 ± 0.30a, a
1% YUPO 59.95 ± 1.55b, a 62.75 ± 0.05a, b 61.90 ± 0.30ab, a
2% YUPO 58.75 ± 0.25c, a 64.70 ± 0.30a, a 62.61 ± 0.03b, a

YPO: yoghurt with peeled oleaster flour; YUPO: yoghurt with unpeeled oleaster flour.
Results are expressed as the mean ± standard deviation (n = 3).
Different letters in the same row and coloumn are significantly different (P < .05).
First letters show differences between the storage times and second letters present the differences between the yoghurt samples.

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H.İ. Öztürk et al.
Fig. 1. Textural profile of yoghurt samples during cold storage. : control yoghurt; : yoghurt with 1% peeled oleaster flour (1% YPO); : yoghurt with 2% peeled oleaster flour (2%
YPO); : yoghurt with 1% unpeeled oleaster flour (1% YUPO); : yoghurt with 2% unpeeled oleaster flour (2% YUPO). The error bars represent standard deviation of means (n=3).
Lower-case letters 3 present the differences between the samples in the same storage time and upper-case letters show differences between the storage times 4 of samples (P < 0.05).
that the addition of over 1% concentration of inulin reduced the con-
sistency of yoghurts. The higher polysaccharide content in 2% con-
centrations may have resulted in the formation of polysaccharide-pro-
tein interactions that are weaker than protein-protein bonds
(Ramchandran & Shah, 2009), therefore, the consistency may have
reduced in yoghurts samples containing 2% concentration of PO and
UPO. Consistency of yoghurt samples showed fluctuation with pro-
gressive time. However, consistency values of 2% YPO, 1% YUPO and
2% YUPO increased at the end of storage compared to the first day. This
finding can be explained by ascended gel firmness during storage and is
in contrast with the report of Aportela-Palacios, Sosa-Morales, and
Velez-Ruiz (2005) who found out a decrease in consistency index of
yoghurts enriched with wheat fiber throughout cold storage.
It was determined that cohesiveness was increased by addition of
PO and UPO on the first day of storage and the highest value was de-
termined in 2% YUPO (P < .05). do Espírito Santo et al. (2012) in-
dicated that the fibers improved the cohesiveness of yoghurts. Ad-
ditionally, Sahan et al. (2015) reported that UPO presented better water
absorption capacity than PO. Therefore, UPO with high water absorp-
tion capacity in the protein matrix may have provided better cohe-
siveness. Relative to the beginning of the storage, cohesiveness values
of yoghurt samples enriched with PO and UPO decreased at the end of
the storage, but they were still higher than the control sample.
The viscosity index of yoghurt samples ranged between 164.73 and
430. 61 g s on the first day of storage (Fig. 1). An escalation in viscosity
index was realized in parallel with the additive concentration used and
the highest viscosity index was determined in 2% YUPO (P < .05).
This could be related with hydroxyl groups in fiber which provides
water interactions by hydrogen bonds and form viscose solution by
absorbing water (Rosell, Santos, & Collar, 2009). Similarly, Aportela-
Palacios et al. (2005) determined that the viscosity of fortified yoghurts
rose out with increased fiber concentration. A reduction in viscosity
index values of all samples was determined after 14 days of storage
because of stability loss (Aportela-Palacios et al., 2005).
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LWT - Food Science and Technology 90 (2018) 620–626
3.6. Microstructure
Microstructure images of yoghurt samples observed with confocal
laser scanning microscopy are shown in Fig. 2. The microstructures of
control sample, 1% YUPO, and 2% YUPO contained a dense protein
network in which the casein micelles formed large aggregates. 1% YPO
also contained large protein aggregates and its protein network was
more heterogeneous by consisting of more serum channels. Moreover,
protein matrix of 1% YPO had a great number of larger serum pores
than those of control, 1% YUPO, and 2% YUPO, Lee and Lucey (2003)
notified that large serum pores caused weak gel structure. Casein mi-
celles in 2% YPO formed the protein network by thin strands and
consequently, the protein structures of 2% YPO presented more open
structure due to the noncompact interactions between casein micelles.
The addition of 2% UPO resulted in a more interconnected network
with smaller serum pores as compared with control. This result was
supported by lower whey separation of 2% YUPO than plain yoghurt.
Additionally, 2% YPO had a more homogeneously dispersed serum
pores with smaller diameter compared to control sample. Sodini,
Remeuf, Haddad, and Corrieu (2004) reported that the reduction in
serum pore size increases the density of protein matrix. This hypothesis
can explain why the viscosity values of yoghurt samples containing 2%
UPO and 2% PO were higher than control.
Although the sizes of serum pores of control sample and 1% YUPO
were approximately similar, the serum phase dispersed more uniformly
in 1% YUPO. It can be attributed to proteins in oleaster fruit which may
have promoted the formation of homogeneous pore structure by in-
creasing hydrophobic interactions in the protein network. 2% YUPO
had larger casein aggregates and more compact network than that of
2% YPO. This microstructural difference may be due to the higher
water binding activity and lower starch content of UPO than PO, that
was reported by Sahan et al. (2015). Sandoval-Castilla, Lobato-Calleros,
Aguirre-Mandujano, and Vernon-Carter (2004) has associated the for-
mation of open structure with starch molecules entering the protein
network. Accordingly, the enhanced interactions between casein
H.İ. Öztürk et al. LWT - Food Science and Technology 90 (2018) 620–626

Fig. 2. Demonstration of the differences in the protein

network of yoghurt samples by confocal laser scanning
microscopy images (on day 14). A: yoghurt with 1%
unpeeled oleaster flour (1% YUPO); B: yoghurt with
1% peeled oleaster flour (1% YPO); C: yoghurt with
2% unpeeled oleaster flour (2% YUPO); D: yoghurt
with 2% peeled oleaster flour (2% YPO); E: control
yoghurt. Scale bar: 10 μm. Red areas indicate protein
network and dark areas show serum phase. (For in-
terpretation of the references to colour in this figure
legend, the reader is referred to the web version of this
article.)

micelles in YUPO owing to low starch content may have been led to the
formation of larger protein aggregates.
3.7. Sensory attributes
Color and appearance, odour, body and texture scores of all yoghurt
samples with oleaster flours were found to be lower than plain yoghurt
(Table 5). Similar results were reported previously for yoghurts en-
riched with some fibers (Guven et al., 2005; Hashim, Khalil, & Afifi,
2009). The changes in the color of the yoghurts (lower brightness, more
redness and yellowness) resulted in decrease in the scores for color and
appearance. No statistical differences were observed between control

Table 5
Sensorial attributes of yoghurt samples.a

Control 1% PO 2% PO 1% UPO 2% UPO

Color and Appearance 5.00 ± 0.00a 4.17 ± 0.98ab 3.83 ± 0.98ab 4.33 ± 0.52ab 3.67 ± 0.52b
Odour 4.67 ± 0.52a 3.17 ± 0.98b 3.50 ± 0.84ab 4.17 ± 0.75ab 4.50 ± 0.55ab
Flavor 4.33 ± 0.52a 2.50 ± 0.84b 3.33 ± 0.52ab 4.17 ± 0.98a 4.33 ± 0.82a
Body and Texture 4.50 ± 0.55a 3.83 ± 0.75ab 3.50 ± 0.55b 4.17 ± 0.41ab 4.17 ± 0.41ab
General Acceptability 4.67 ± 0.52a 3.00 ± 0.89c 3.33 ± 0.52bc 4.33 ± 0.52ab 4.50 ± 0.55a

YPO: yoghurt with peeled oleaster flour; YUPO: yoghurt with unpeeled oleaster flour.
Different letters in the same row are significantly different (P < .05).
a
The results were given for 14 days.

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H.İ. Öztürk et al.
sample and 2% YUPO with regards to flavor and general acceptability
scores, while these scores were lower in 1% YPO and 2% YPO. The
addition of 2% UPO supported the maintenance of sensory properties
similar to those of conventional yoghurt. However, this result is in
contrast with the reports of previous studies in which flavor and general
acceptability scores were diminished by fiber addition to yoghurt
(Hashim et al., 2009; Staffolo, Bertola, & Martino, 2004).
4. Conclusion
Based on the findings obtained from this study, 2% PO and UPO
supplementation to yoghurts statistically accelerated the fermentation
time and improved the water holding capacity during cold storage
thereby decreasing syneresis. Enrichment with PO and UPO enhanced
cohesiveness and viscosity index of set-type yoghurts, however, it re-
duced firmness and consistency compared to control sample. 2% YUPO
represented better microstructure with regards to smaller serum pores
and thus the increased interactions between protein aggregates com-
pared to control. Unexpectedly, yoghurts fortified 2% UPO had good
general acceptability scores similar to plain yoghurt. Based upon higher
total phenolic content and antioxidant activity, shorter fermentation
time, higher water holding capacity and viscosity index, better micro-
structural characteristics, incorporation of 2% unpeeled oleaster flour
into yoghurt can be recommended to developing a satisfactory alter-
native for further new product studies and improving functionality of
yoghurt.
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