Yogurt Paper

International Journal of Biological Macromolecules 263 (2024) 130428
Contents lists available at ScienceDirect
International Journal of Biological Macromolecules

journal homepage: www.elsevier.com/locate/ijbiomac
Review
Perspectives on the yogurt rheology

Didem Sözeri Atik a, b, *, **, Hale İnci Öztürk c, Nihat Akın d
a
Tekirdağ Namık Kemal University, Department of Food Engineering, Tekirdağ, Turkey
b
University of Wisconsin-Madison, Department of Food Science, Madison, WI, USA
c
Konya Food and Agriculture University, Department of Food Engineering, Konya, Turkey
d
Selçuk University, Department of Food Engineering, Konya, Turkey
A R T I C L E I N F O A B S T R A C T
Keywords: The oral processing of yogurt is a dynamic process involving a series of deformation processes. Rheological
Structural properties knowledge is essential to understand the structure and flow properties of yogurt in the mouth and to explore its
Acid gels relationship with sensory perception. Yogurt is rheologically characterized as a non-Newtonian viscoelastic
Mouthfeel
material. The rheological properties of yogurt are affected by many factors, from production to consumption.
Non-Newtonian
Rheological methods
Therefore, rheological measurements are widely used to predict and control the final quality and structure of
Tribology yogurts. Recent studies focus on the elucidation of the effects of cultures and processes used in production, as
well as the design of different formulations to improve the rheological properties of yogurts. Moreover, the
science of tribology, which dominates the surface properties of interacting substances in relative motion to
evaluate the structural sensation in the later stages of eating in addition to the rheological properties that give the
feeling of structure in the early stages of eating, has also become the focus of recent studies. For a detailed
comprehension of the rheological properties of yogurt, this review deals with the factors affecting the rheology of
yogurt, analytical methods used to determine rheological properties, microstructural and rheological charac
terization of yogurt, and tribological evaluations.
1. Introduction through the implementation of several strategies: increasing the milk

solids/protein content, thermal treatment and homogenization of the
Rheology is a field of physics that focuses on the deformation and milk base, and utilization of commercial starter cultures [10].
flow of complex fluids and soft materials, such as foods, when subjected Structural analysis of yogurt includes the evaluation of textural,
to external forces. These forces can either occur naturally, such as rheological, and microstructural properties. The evaluation of gel (set-
gravitational or interaction forces that hold a structure together, or they type yogurt) and liquid (stirred-type yogurt) characteristics of yogurts is
can be intentionally applied during industrial processes, use, or con conducted by textural and rheological analysis methods [11]. In addi
sumption. Subsequently, rheological processes play crucial roles in the tion, in recent times, novel methodologies like tribology have been
fundamental, technical, and sensory aspects of dairy products such as employed to simulate the interaction between the tongue and palate
yogurt, which is among the most well-known and oldest fermented dairy inside the mouth to develop a better understanding of the contributions
products that are consumed worldwide [1,2]. of structure to sensory characteristics [12,13]. Moreover, the micro
Yogurt is a partially solid food that contains a protein network with structural analysis of yogurt comprises two components: an examination
integrated fat globules in its microstructure [3,4]. The microstructure of microstructural images to investigate the detailed structure and
and rheological attributes of yogurt play a crucial role in determining conducting microstructural assays, including those involving EPS (exo
the quality of the product since they are closely associated with desirable polysaccharides produced by bacteria) and those used to determine the
characteristics such as flowability, firmness, and smoothness [5,6]. average diameter of fat globules/particles [11].
Subsequently, the structural features of yogurts play a crucial role in Accordingly, the main objective of this review is to provide a
determining consumer acceptance [7–9]. In the dairy industry, consis comprehensive understanding of the influencing factors of yogurt
tent production of yogurt with the desired structure is accomplished rheology, starting from the ingredients and processing steps to the
* Corresponding author at: Department of Food Science, University of Wisconsin-Madison, Madison, USA.
** Corresponding author at: Department of Food Engineering, Tekirdağ Namık Kemal University, Tekirdağ, Turkey.
E-mail addresses: [email protected], [email protected] (D. Sözeri Atik).
https://doi.org/10.1016/j.ijbiomac.2024.130428
Received 26 November 2023; Received in revised form 21 February 2024; Accepted 22 February 2024
Available online 23 February 2024
0141-8130/© 2024 Elsevier B.V. All rights reserved.
D. Sözeri Atik et al. International Journal of Biological Macromolecules 263 (2024) 130428
techniques for evaluating the yogurt rheology and structure. For this [20,21]. In this context, milk is evaporated or ultrafiltered to increase
purpose, the review begins by providing a concise overview of the fac the total solid content to provide the desired structure in the final
tors that affect yogurt rheology. Then, it discusses the analytical product. In addition, fat [22], skimmed milk powder [23], and various
methods that are used to understand the rheological characteristics of protein concentrates [24] are used to increase the total solid content of
yogurt. The last section offers important information regarding micro yogurt milk. They are aimed to improve the rheological properties of
structure, as well as current details on tribology perspectives of yogurt. yogurt by reducing syneresis with these approaches.
The protein content of yogurt milk plays an important role in
2. The factors that affect yogurt rheology rheology since yogurt has a gel structure consisting of protein nets.
Casein is the main protein found in milk. As a result of colloidal calcium
From a rheological point of view, milk, a Newtonian liquid, turns phosphate dissolution (maximum dissolution pH 5.0) in casein micelles
into yogurt, a non-Newtonian liquid, at the end of the fermentation [14]. due to decreasing pH values in milk, caseins become irregular and form a
The gel structure of yogurt occurs as a result of the formation of protein gel network structure [25]. The three-dimensional network structure
aggregates depending on the instabilities of the casein micelles by the formed between the protein forms the acid gel structure. This network
developing acidic conditions [15]. For this reason, the three- structure can be modified by protein and fat content. To increase protein
dimensional structure formed is defined as an acidic gel. The rheolog interaction, extra protein supplementation to yogurt milk will be a
ical properties of yogurt are affected by various factors, from the starter healing approach [26]. For this purpose, studies have focused on forti
cultures used to storage conditions [16]. In this context, the critical fying yogurt milk with whey components and sodium caseinate [24].
parameters affecting the rheological properties are presented by cate Hashim et al. [27] have reported that whey protein isolate improves
gorizing in Fig. 1. This section evaluates each parameter separately. rheological properties by increasing the water-holding capacity of yo
gurts. Besides, it was determined that the addition of whey protein to
milk improves the rheological properties since the cross-links estab
2.1. Yogurt milk composition lished between caseins and whey proteins create a denser protein
network [28]. A similar situation has been reported for the addition of
Total solids content is known to be important in yogurt rheology sodium caseinate [29]. In general, literature studies show that whey
[17]. Wu et al. [18] have reported that increasing total solids content protein or sodium caseinate is added to improve the rheological prop
increases the apparent viscosity of yogurt. Penna, Converti, and De erties of low-fat and reduced-fat yogurts [30].
Oliveira [19] examined the rheological properties of stirred yogurts with Milk fat contributes to sensory properties such as mouthfeel, texture,
a total solid content ranging from 9.3 to 22.7% and observed that the aroma, and lubricity in yogurt [14]. The fat globules attached to the
consistency index (K) increased, and the flow behavior index (n) proteins, which are formed by the ability of milk proteins to coat the
decreased with increasing dry matter ratios. For yogurt, the increase in surface of the fat globules, help the formation of the yogurt network
the consistency index can be considered as an improvement in the [31]. Therefore, the covering of homogenized fat globules with casein
rheological properties. It has been reported that increased milk total dry facilitates protein-protein interactions, and this way of trapping the fat
matter content between 12 and 20% improves the consistency of yogurt in the protein network increases the strength of the protein gel [32]. In
[16]. From the various studies, it is seen that the dry matter of yogurt rheological measurements, it has been reported that the elastic modulus
milk is standardized as at least 14% to obtain the desired structure
Fig. 1. Variable factors contributing to yogurt rheology (created with BioRender).
2
(G′) is related to the amount of oil, and the increased amount of oil researchers observed a coagulum in the form of dispersed casein flakes
causes higher G′ values, which indicates a more rigid structure [14]. with a fragile and heterogeneous structure in camel milk yogurts.
Accordingly, low-fat content results in a poor gel structure in yogurts. Because of these properties, it has been suggested that camel milk should
Lussier et al. [22] investigated the effect of milk fat at rates ranging from be used in the production of drinking yogurt rather than yogurt [51].
0 to 3.9% on the rheological properties of yogurt. These researchers Recently, due to some health advantages (lactose- and cholesterol-
detected a stronger, more viscous gel structure with increasing oil con free) and increasing consumer demand, the replacement of original
tent. Consumers have recently shown a demand for low-fat yogurts due milk by plant milk in various fermented milk products has gained mo
to health concerns [33]. Reducing the fat in yogurt causes structural mentum. In this context, yogurt has often been the subject of studies as a
defects by causing low viscosity. As mentioned before, whey proteins fermented product in which several vegetable milk species are experi
have the potential to be used as a fat substitute to overcome this situa enced [41]. Mechanisms of caseins and vegetable proteins to form
tion. Moreover, food ingredients such as inulin [34], glucomannan [35], yogurt gel are similar [15]. This mechanism can be explained by (i) the
various hydrocolloids [36] can be added to the formulation of yogurt use of sugar by lactic acid bacteria to form lactic acid, (ii) triggering the
milk, which strengthen the protein network and increase the apparent formation of protein aggregates with a decrease in net load and repulsive
viscosity. Renan et al. [37] determined that yogurt gels with high lactic forces depending on the decreasing pH values, and (iii) the formation of
acid content showed higher G′ and breaking strength. a three-dimensional gel structure [52]. However, it has been reported
Yogurt bacteria break down the milk sugar lactose during milk that grainy structure, low consistency, and high serum separation occur
fermentation to form lactic acid. The resulting lactic acid causes the in yogurts due to poor gelling and solubility properties of vegetable
development of acidity in yogurt. This triggers the separation of serum proteins other than soy protein [15]. Moreover, the absence of lactose,
from yogurt [38]. However, since serum separation is considered a which is required for the development of lactic acid bacteria, in vege
physical defect in yogurt, it should be kept as low as possible. This sit table milk requires the addition of an additional sugar source to the
uation can be controlled by minimizing the development of acidity yogurt formulation [53]. Zhao, Bhandari, Gaiani, and Prakash [54] re
during the storage of yogurts. ported that a better gel structure (higher G′) was obtained by increasing
the protein and sugar concentration in plant-based yogurts. In addition,
2.2. Milk source these researchers determined that the increased fat content increased
the G′ value of the yogurt gel since the fat embedded in the protein
Depending on the compositional changes described above, the milk matrix served as a filler. Besides these, other ingredients in plant milk
source used in yogurt production affects the rheological properties of the can affect the gel texture of yogurt. In this regard, starch content is
final yogurt. Generally, bovine milk is used in yogurt production [14]. In substantial. Brückner-Gühmann, Banovic, and Drusch [55] reported that
addition, sheep, goat, buffalo, and camel milk or their mixtures with starch increased gel hardness (G′) and water-holding capacity by acting
cow’s milk are optionally used in yogurt production [39,40]. Due to the as a filler in the yogurt protein network. Further studies are needed to
increasing demand for plant-based foods based on the lifestyle and determine the effect of other components in the matrix of vegetable milk
health concerns of consumers, vegetable milk has also been used as an types on the yogurt gel network structure. It is clear that the low rheo
alternative in yogurt production [41]. logical properties of plant-based yogurts will be improved by changing
It has been reported that the casein content of milk has a significant the yogurt formulation. In the meantime, focusing on studies to increase
effect on the rheological properties of the gel structure [42]. The lower the solubility of vegetable proteins will add value to these products.
αs-casein content in goat milk compared to cow and sheep milk results in
lower viscosity in acid gels, showing weaker structural properties [43]. 2.3. Starter cultures
Nguyen, Afsar, and Day [44] reported that sheep milk with a high total
solids content provided a denser protein network and gel firmness to the Yogurt is a fermented milk product produced by Streptococcus (S.)
yogurt gel. Additionally, the high casein and fat content of sheep milk thermophilus and Lactobacillus delburieckii subsp. bulgaricus (L. bulgaricus)
compared to other milk makes the rheological properties of yogurt among the lactic acid bacteria [16]. Besides these, yogurt is also a good
produced from sheep milk superior [45]. Nguyen et al. [44] reported vehicle for delivering probiotic bacteria in the food matrix to consumers
that sheep milk gel had a much higher storage modulus (G′) than goat [56]. The selection of the appropriate starter culture in yogurt produc
milk gel at the end of the fermentation of yogurt milk, showing a tion is essential for the aroma, acidification, texture, and stability of the
stronger gel structure. final product [14]. Many strains of S. thermophilus and L. bulgaricus are
Al-Bedrani, Alkaisy, and Rahi [46] determined the best rheological used commercially in yogurt production [57]. However, in yogurt pro
properties (dense protein network interaction, reduced serum separa duction, the selection of strains capable of producing acetaldehyde and
tion) in yogurts produced from buffalo and sheep milk as compared to exopolysaccharide (EPS) are important in terms of aroma and textural
cow and goat ones due to high protein and dry matter contents. In properties, respectively. Although L. bulgaricus has been shown to be
another study, Jumah, Shaker, and Abu-Jdayil [47] observed the highest responsible for acetaldehyde production, there are also contrary opin
viscosity value in yogurts produced using sheep milk depending on total ions [58]. This shows that acetaldehyde production varies according to
solids and protein content, followed by goat, cow, and camel milk yo the strain type. Similarly, it has been reported that EPS production also
gurts. Furthermore, these researchers did not detect a significant change varies depending on the strain, and some strains of both L. bulgaricus and
in the viscosity of yogurt produced from camel milk during gelation. In a S. thermophilus have high EPS-producing ability [59].
different study, the gel strength of yogurts produced by mixing cow milk Polysaccharides play an active role in consistency and viscosity,
with camel milk has been reported to decrease [39]. This may be due to especially in stirred-type and reduced-fat yogurts [60]. EPSs produced
the lower κ-casein content and the deficiency of β-lactoglobulin in camel by yogurt bacteria in milk media contain mostly galactose and glucose,
milk [48]. It has been reported that the interactions of κ-casein and together with small amounts of mannose, fructose, rhamnose, arabinose,
β-lactoglobulin among themselves and with other milk proteins are xylose, or N-acetylgalactosamine, individually or in combination [61].
important in the formation of yogurt gel [49]. Accordingly, the lack or Although significant differences can be observed between various
absence of these interactions may be an inhibitory force in camel milk strains of yogurt bacteria in the amount of polysaccharide produced, this
coagulation. The low κ-casein content of camel milk has been reported variation has been reported to present no effect on the apparent viscosity
to cause problems in the production of camel milk yogurt [14]. Attia, of the final product [62]. Therefore, polysaccharide type may be more
Kherouatou, and Dhouib [50] reported that there was lower lactic acid effective on the rheological properties of yogurt rather than the quantity
fermentation in camel milk due to the culture activity, and the coagulum of EPS.
formed in camel milk was quite different from cow milk. Besides, these It has been reported that the mixed culturing of EPS-producing and
3
non-producing strains promotes EPS biosynthesis and provides the op Table 1
portunity to produce EPS of different structures and types [59]. Thus, Effects of process conditions on yogurt rheology.
this mixed culture approach has come to the fore to improve the rheo Process Changes Reference
logical properties of yogurts [63]. In addition, the use of EPS-producing
Set-type yogurts
probiotic strains in yogurt production aims to improve both the rheo Increased fermentation Increase in G′ Haque, Richardson, and
logical properties and the health properties of the final product (P. [64]). temperature (37-46̊ C) Morris [79]
Zhao and Liang [65] observed higher apparent viscosity, cohesiveness, Increased fermentation Increased viscosity, Paz-Díaz, Mora-Vergara,
and consistency in yogurts produced with EPS producer probiotic time reduced syneresis Navarro-Arana, Navarro,
and Pacheco-Valderrama
L. plantarum MC5 compared to yogurts produced with commercial [80]
yogurt cultures. It has been reported that EPS occurring in yogurts Storage temperature for No impact on G′ Renan et al. [37]
contributes to adhesiveness rather than firmness and flexibility, which stirred-type yogurts
are affected by the protein matrix [14]. In other respects, it has been Increased Reduced gel firmness and Nguyen et al. [76]
homogenization storage modulus
suggested that the use of EPS-producing strains in yogurt production
pressure (80-160 bar)
causes lower acetaldehyde content compared to non-EPS-producers Cooling temperature 0 ◦ C showed a more Oraç and Akın [81]
[66]. Regarding this, the combined usage of EPS-producing and non- (− 10 ◦ C, − 5 ◦ C, 0 ◦ C, homogeneous protein
EPS-producing strains will be a solution-oriented approach in terms of 4 ◦ C) matrix and gel firmness
both structure and flavor development in yogurt.
Stirred-type yogurts
2.4. Yogurt type (set or stirred) Increased fermentation Increase in G′ Haque, Richardson, and
temperature (37-46̊ C) Morris [79]
Its combination with heat
Yogurts can be divided into two categories: set-type and stirred-type. High hydrostatic
treatment (85 ◦ C for 30 Penna, Gurram, and
pressure (676 MPa for
Set-type yogurts are coagulated and fermented directly in the final min) increased gel Barbosa-Cánovas [82]
5 min)
package without any interference while following the fermentation in a firmness
large tank, and the stirred-type yogurts are mixed, pumped, packaged, High-intensity
Increased viscosity,
thermosonication Ragab et al. [83]
and stored [57]. The two primary types of yogurts, set-type, and stirred- pretreatment of milk
reduced syneresis
type, exhibit distinct rheological characteristics. Set-type yogurt is Nöbel, Protte,
Sonication (35 kHz)
characterized by a firm and gel-like consistency [67]. The gel network in Larger casein micelles Körzendörfer, Hitzmann,
during fermentation
set-type yogurt is undisturbed. Due to their firm structure, set-type yo and Hinrichs [84]
Tubular heat exchanger
gurts typically exhibit lower whey separation compared to stirred-type Firmer yogurt gel Lussier et al. [22]
for cooling step
yogurts [68]. On the other hand, stirred-type yogurts have lower vis Plate heat exchanger for
cosity and firmness and smoother texture compared to set-type yogurts Reduced syneresis Lussier et al. [22]
cooling step
[69]. After fermentation, the yogurt gel is broken and stirred to achieve
a homogeneous consistency. The gel network in stirred-type yogurt is
disrupted due to the stirring process, leading to a different mouthfeel 72 ◦ C/30 s, 85 ◦ C/5 min, and 95 ◦ C/5 min, Hovjecki, Miloradovic, Rac,
compared to set-type yogurts. It has been determined that the viscosity Pudja, and Miocinovic [78] observed that 85 ◦ C/5 min treatment gave
of stirred-type yogurts decreases with mixing or pumping processes but the firmer yogurt gel structure due to the formation of larger casein
increases again during storage [4]. Moreover, the stirring process causes micelle structure.
higher syneresis than set-type yogurts due to the disruption of the gel Fermentation conditions such as temperature and duration can in
network during this process [70]. For enhancing the viscosity and gel fluence the rheological properties of yogurt. Haque et al. [79] reported
strength of stirred-type of yogurts, microbial transglutaminase [67], that the G′ value increased with increasing fermentation temperature.
increased protein concentration [71], hydrocolloid (such as pectin, guar Lee and Lucey [85] determined that the lower fermentation temperature
gum, gelatin) addition [72], and optimized fermentation conditions combined with the high preheating process applied to milk resulted in a
[73] have been used. In addition, stirred yogurts are also produced with higher increase in storage modulus and apparent viscosity values.
fruit supplementation. Stirred yogurts containing fruits exhibit distinct Therefore, the effect of high preheating at high temperatures on
viscoelastic properties. The presence of fruit particles in yogurts can rheology seems to be more effective compared to the fermentation
increase the storage modulus (G′), indicating a stronger gel structure temperature. In this case, from both economic and technological views,
[74]. production processes should be designed by considering the relationship
of all parameters rather than just one parameter. Long fermentation time
2.5. Processing conditions has been reported to improve yogurt viscosity [80]. Similarly, [86] re
ported that the extended fermentation time resulted in a thicker gel
Various production parameters, including the heat treatment con structure with high water-holding capacity due to the tight arrangement
ditions of milk, homogenization, and fermentation conditions, influence of the protein network structure. For these reasons and depending on the
the rheological properties of yogurt. The effects of processing factors on culture activity, the fermentation time in commercial yogurts ranges
yogurt rheology are summarized in Table 1. Milk homogenization has a from 4 to 12 h.
significant impact on the rheological properties of yogurt. Homogeni
zation breaks down fat globules in milk, leading to increased firmness,
viscosity, and water-holding capacity in yogurt [75]. However, Nguyen, 2.6. Storage
Ong, Kentish, and Gras [76] reported that increasing homogenization
pressures decreased gel firmness. Therefore, by modulating the ho Yogurt is a perishable cultured dairy product with a shelf life of up to
mogenization parameters, manufacturers can achieve the desired 30 days under cold storage conditions [16]. Textural properties, which
rheological properties in yogurt, enhancing its quality and consumer are important quality criteria for consumers, may change during stor
acceptability. In yogurt production, the heat treatment of milk before age. Renan et al. [37] have reported that during cold storage of yogurts,
fermentation can denature whey proteins, allowing them to participate some changes, including the rearrangement of casein networks, occur,
in gel structure and rheological properties. Jørgensen et al. [77] have which causes an increase in firmness and viscosity. Lussier et al. [22]
reported that increased whey protein denaturation increases the storage found that the firmness of yogurt increased significantly, especially
modulus and yields firmer gels. Among heat treatment conditions of between the 7th and 21st days of storage time. Besides, these researchers
4
observed that the apparent viscosity increased with advancing storage completely reversible once shear is eliminated [91]. The time-dependent
time. This may have been caused by the rearrangement of the protein flow behavior properties of yogurts were determined using a hysteresis
network structure due to post-acidification and EPS produced by yogurt loop in the flow curves and shear stress decay [100]. The presence of
bacteria [87] or the reduction of hydrophobic interactions [88] during hysteresis in the flow curve (shear stress versus shear rate) indicates that
storage. Technically, these changes observed in protein networks during shearing at a high shear rate alters the structure of the material. During
storage in yogurt are called rebodying. Renan et al. [37] have reported the shearing cycle, the area of the hysteresis loop relates to the degree of
that storage temperature (4-20 ◦ C) in stirred-type yogurts does not affect structural degradation [101,102], and the flow curves of the hysteresis
the rheological properties if over-acidification is prevented. loop are fitted to flow behavior models, such as the Herschel Bulkley and
Power Law models [103]. On the other hand, in the shear decay test,
3. Analytic methods on the measurement of rheological samples are subjected to a constant shear rate over time, and the shear
characteristics in yogurt stress is recorded as the response [89].
Small amplitude oscillatory shear (SAOS) and large amplitude
Understanding the mechanical and rheological properties of oscillatory shear (LAOS) are the two types of dynamic oscillatory shear
different dairy products is essential for the design of flow procedures for tests. The primary distinction between them is that SAOS tests are
quality control, the prediction of storage and stability evaluations, and conducted in the linear viscoelastic region and are supported by a solid
the development of texture. Also, the selection of an instrumental theoretical foundation, whereas LAOS testing is conducted outside of
technique for determining the rheological characteristics of a dairy this region [104]. Fig. 2 illustrates the strain sweep test conducted at a
product depends on the properties of the product (i.e., whether it is constant frequency. The sweep test has the capability to ascertain both
liquid, semi-solid, or solid) and the goal of the measurement [89]. The the linear and nonlinear viscoelastic area [104,105]. Various rheological
following methods can typically be used to evaluate the structural parameters of yogurts have been determined during and after the gel
properties of yogurt: viscometry, penetrometry, small-amplitude oscil formation process (or fermentation) using oscillatory tests carried out
latory rheology, and large-amplitude oscillatory shear [90]. over time at constant strain or stress, frequency, and temperature
In the food industry, viscometers are frequently used to describe the without causing disruption to the structure of yogurt gel network. It is
flow properties of yogurt samples [90]. Due to the fact that a viscometer possible to differentiate the structure of ‘strong’ and ‘weak’ gels using
with a spindle only measures an “apparent” viscosity at one spindle small deformation oscillatory tests. Calculating storage modulus (G′) and
speed that is empirically chosen as a “consistent” reading after some loss modulus (G″) in the linear viscoelastic region and phase angle
shearing period, it is impossible to transform the empirical results into characterizes the viscoelasticity and structure of yogurt gels [91,104].
more fundamental data of stress and shear rate when using this mea Using frequency sweeps, one may determine the sort of gel generated
surement method [90]. In addition, because these experiments are often in the sample. Yogurt can be considered a physical gel because the dy
conducted at only one or two rotating speeds, the results can result in a namic moduli rise with frequency and G′ > G″ throughout the entire test
lack of understanding of yogurt’s flow behaviors, which can lead to [91,107]. The physical properties of yogurt protein gels are influenced
inaccurate observations regarding its processing and sensory behaviors. by a balance between attractive forces like hydrophobic attractions and
Flow behaviors in yogurt can be highly complex and must be examined various types of cross-links and repulsive forces such as electrostatic
at a variety of shear rates for a comprehensive knowledge of the flow repulsions, which are key in defining the structure of gel [74].
behavior profile [91]. Large amplitude oscillatory shear (LAOS) has developed as an
Penetrometer is also used to determine the gel firmness of yogurt appropriate method for assessing non-linear rheological properties due
samples, and results are revealed as the displacement of penetration at a to the fact that amplitude and frequency can be separately controlled to
constant speed [92]. Therefore, the penetrometer reading is inversely alter both the strength and timescale. LAOS can simulate a variety of
associated with the firmness of samples [93]. However, a penetrometer mechanical situations, including food manufacturing, expanding the
is a unidimensional instrument that mainly evaluates the firmness of scope of food rheology experiments from “theoretical estimation” to
samples and is frequently an unreliable indicator of other complicated
texture characteristics [94].
Rheological measurements are regarded as the response at the
macroscopic level to the microscopic features of food [95]. These ex
periments ensure complete information about the structure of yogurt,
but to perform these tests, it is essential to retain the structure of the
yogurt when preparing and loading samples. This might be challenging
for set-type yogurts, as taking them from the containers typically causes
the gel to break [91,96], because casein strands and whey protein ag
gregates form a particulate, heterogeneous, and three-dimensional
network in yogurt [97]. Generally, flow tests, small amplitude oscilla
tory tests, and large amplitude oscillatory shear tests are used as
different rheological methods to characterize the structure-related
properties of yogurt samples [98].
The flow properties of yogurts are characterized by non-Newtonian
behavior, and all yogurts can be characterized as shear-thinning sub
stances whose viscosity decreases with increasing shear rate [91]. Since
the shear rate affects the viscosity of non-Newtonian substances, the
word “apparent viscosity” is used to describe viscosity more precisely. It
refers to the viscosity at a specific shear rate. When evaluating the
rheological properties of semi-solid foods, such as yogurt, the apparent
viscosity at a shear rate of 50 s− 1 (η50) is commonly employed. This
value corresponds to the shear rate produced by the tongue to the palate
in the mouth [99]. Additionally, yogurt, as a semi-solid dairy product, Fig. 2. The schematic representation of the strain sweep test at a constant
has a yield stress value, which is the minimum stress to ensure the frequency.
sample flows. Moreover, yogurt products exhibit thixotropy that is not The figure is reproduced with permission from Hyun et al. [106].
5
“close to real measurement” [106]. Furthermore, Lissajous curves, sample preparation because only one dye is utilized [119]. Besides
Fourier transform rheology, stress decomposition, computation of these, scanning (SEM) or transmission electron microscopy (TEM) per
viscoelastic moduli, decomposition into characteristic waveforms, and mits viewing of the fractal cluster level and major component particles;
analysis of Fourier transform rheology parameters are some methods for however, the sample preparation technique may cause errors and must
analyzing LAOS data [108]. In a recent study by Erturk, Bonilla, and be carried out with care ([98]; Miloslav [125,126]). However, cryo-SEM
Kokini [109], the LAOS rheological behavior of yogurt samples, which is regarded as the optimal technology for analyzing dairy products with
have different fat percentages, was evaluated to provide further insights a high water and fat content, such as yogurt [127,128]. In the past,
into the distinctive structure of yogurt. It was found that the structure techniques, such as gray-level co-occurrence matrix (GLCM) analysis
and deformation properties of the samples are linked to their structure. and fractal analysis, have been effectively employed to examine the
Also, the outcomes of the study would have the potential to facilitate the microstructure of protein networks in yogurts [129,130]. On the other
development of more regulated designs for yogurt products and pro hand, when evaluating the automated objective description of yogurt
duction processes, accommodating the desired modifications in formu microstructure, the findings of Skytte et al. [120] indicate that utilizing
lation and ingredients as desired by consumers. texton-based image descriptors may be advantageous compared to
traditional descriptors like fractal analysis and GLCM.
4. Yogurt microstructure, rheology, and texture The microscopy techniques facilitate the analysis of foods at a
microscopic scale, resulting in a greater understanding of how their
The microstructure of foods is essential for comprehending the microstructure influences their rheological properties (Fig. 3). For this
spatial configuration of food components and their interconnections at purpose, various models have been proposed to characterize the
sub-100 μm scales, and it gives descriptive data about their physical microstructure of yogurt, as the rheological properties of yogurt are the
state [110,111]. Therefore, the microstructure and rheology of yogurts macroscopic representation of the characteristics seen at the micro
are crucial attributes due to their association with processing conditions, scopic level of food [95,131]. For instance, a micro-rheological model
nutritional bioavailability, and functional characteristics [5,112]. was employed to elucidate the viscosity of yogurt by considering its
Yogurt microstructure refers to the organization of protein, fat, and microstructure, including the features of microgels, such as elasticity,
other substances in a three-dimensional gel network that is generated network rigidity, and porosity, as well as the ability of protein aggre
and maintained throughout fermentation and has a substantial impact gates to undergo interactions, breakage, and reforms [132]. Also,
on the rheology and texture of the sample [113]. Furthermore, the acid structural-kinetic models, which establish a relationship between time-
gels are composed of a coarse network of casein particles accompanied dependent rheological properties and alterations in the underlying of
by clusters, chains, and strands. The network has holes or empty areas microstructure, have been developed and used to explain the structure
that limit the aqueous phase. The presence of fat globules in yogurt of acid milk gels [133,134].
products makes it difficult to see pores and strands. On the other hand, According to the International Standard Organization, food texture
large casein clusters can be seen in stirred yogurt, which was likely refers to the mechanical, tactile, visual, and auditory aspects of a
produced by shearing and collisions that occurred during mixing product that may be perceived by our senses, including its rheological
[74,85]. Many parameters of the process can greatly influence the (flow and deformation) and structural (shape and surface) characteris
microstructure of yogurt gel, such as ingredient composition and tem tics [142,143]. The physical characteristics of yogurt gels, such as their
perature (Table 2). Changes in the appearance of microscope images, stiffness and permeability, as well as the rearrangement of protein
which are often defined by the amount of protein interconnection, size particles within the gel network and the breakdown of structure in
distribution of pores, and tortuosity, can reveal these impacts [114]. stirred-type yogurts, are crucial elements that affect the physical and
To evaluate the microstructure of yogurt gel, a wide number of structural characteristics of yogurts [74]. Consequently, obtaining
methods are available, providing the opportunity for various scales of knowledge about the mechanisms underlying the generation of texture
observation from μ to a few cm [68,98,115]. For instance, laser in yogurts and the influence of processing conditions on texture devel
diffraction spectrometry and sieving technologies provide accessibility opment is crucial in enhancing the overall quality of yogurt [90]. For
to a wide variety of particle sizes and sample dispersion with statistically instance, the dimensions and dispersion of particles in yogurt, such as fat
precise results. On the other hand, observation of the structure and globules and protein, play a role in determining its texture. Rheological
counting approaches utilizing microscopy and image analysis have a approaches can help in the characterization of these particles and
lesser statistical accuracy but offer high-resolution features and a great facilitate the comprehension of their influence on the overall texture
deal of structure-related information [98,116,117]. The laser diffraction [24]. Consequently, the impact of milk base composition, starter culture,
spectrometry technique depends on the ability of suspended particulates and process technique on yogurt texture varies in terms of range and
to scatter a given beam of light according to their sizes, refractive significance and is contingent upon the methodologies employed for
characteristics, and dispersant refractive qualities [117,118]. texture evaluation. Indeed, several textural characteristics can be
The latest developments in microscopy offer new possibilities for assessed using sensory analysis, physical tests, such as viscoelasticity,
investigating the microstructure of foods [119]. For instance, confocal viscosity, and water holding capacity (WHC), or microscopy techniques
scanning laser microscopy (CLSM) has become the standard measure [4].
ment method for fermented dairy products since the microstructure of
the protein network in yogurts determines crucial physical qualities. 5. Tribology perspectives
Because using CLSM to image this protein structure has yielded favor
able findings [120]. In addition, other light microscopy techniques have Tribology is the analysis of the behavior of lubrication, friction, and
also been used to observe the microstructure of dairy products, such as wear between two surfaces that are sliding [144]. Although standard
fluorescence microscopy, polarized light microscopy, and bright field rheological analyses make it relatively simple to evaluate the flow
light microscopy [121]. Moreover, coherent anti-stokes Raman scat profiles, yield stresses, and fracture behavior of solids, there are other
tering (CARS) microscopy has been applied to the scanning of the features that regular rheological testing cannot quantify. In the final
structure of various foods and permits label-free images, as the approach steps of mastication, when food is ready for swallowing, mechanical
is tailored to a particular vibrational mode of the component of interest parameters correlate poorly with food texture. One of the primary goals
[122,123]. In addition, the two-photon excitation (2 PE) microscopy of food tribological experiments is to establish a correlation between
enables the imaging of deep tissue [124]. However, the 2 PE method can food texture and mechanical friction data [104]. As a result, Chen and
be conducted concurrently with CARS microscopy, and it enables the Stokes [145] reported that tribology and rheology can present distinct
acquisition of photos with the same quality as CLSM with minimal parts of oral behavior and associate with particular sensory qualities or
6
Table 2
The effects of different factors on the microstructural and rheological properties of yogurts.
Yogurt Instrument for Microstructure observation Rheological effect Reference
microstructural
analyses
Set-type yogurts
Low-fat yogurt samples from cow, CLSM and cryo- Goat yogurt showed a porous structure and The weaker structure of the goat yogurt is Nguyen et al. [44]
goat, and sheep milk SEM low-dense protein network; it is followed proven by the storage modulus value,
by cow yogurt. Sheep yogurt samples which was substantially lower after the
exhibited the most compact and fermentation than that of cow and sheep
interconnected protein structure. yogurt samples.
Buffalo yogurt is produced at CLSM and cryo- The microstructure of yogurt fermented at The fermentation temperature did not Nguyen, Ong, Kentish,
different fermentation SEM 37 ◦ C was compact, but the microstructure significantly alter the flow behavior index and Gras [136]
temperatures (37, 40, and 43 ◦ C) of yogurt fermented at 43 ◦ C was more and consistency coefficient of the samples
porous. during storage.
Buffalo yogurt produced from milk CLSM and cryo- Two treatments resulted in a decrease in The homogenization process provided a Nguyen et al. [76]
which homogenized at different SEM the dimension of fat globules, an increase yogurt gel with a higher storage modulus
pressure values (80 or 160 bar) in the particular surface area of fat, and the (G′), firmness, and flow behavior index
development of fat clusters and protein with a lower hysteresis area. However, 160
coating. bar homogenization pressure led to a
decrease in gel firmness and G′ compared to
a pressure of 80 bar.
Low-fat, set yogurt produced with Cryo-SEM Control yogurt has a more homogenous A high value of G′ was observed in yogurts, Prasanna et al. [64]
two different exopolysaccharide microstructure with a compact form. including EPS-producing strains, especially
(EPS) producing Bifidobacterium Yogurts with Bifidobacterium longum subsp. in CCUG yogurts.
strains infantis CCUG 52486 had a well-defined
porous pattern with higher pore diameters.
Yogurts produced from a mixture of SEM The addition of camel milk to bovine milk A decline in gel strength with the addition Kamal-Eldin et al. [39]
camel milk and bovine milk produced casein micelles that were bigger of camel milk led to a reduction in viscosity
and less recognized. The pure bovine and G′, G″.
yogurt produced small and distinct
particles, but the addition of camel milk
produced aggregates that were bigger, less
rigid, and with less visible inter-particle
gaps.
Yogurt manufactured by heating Fluorescence No significant variations in microstructure The G′ profiles of yogurt samples produced Ozcan, Horne, and
milk to varying pH levels (6.2, microscopy were identified as a result of the various from milk heated to a pH of 6.7 were Lucey [137]
6.7, and 7.2) treatments. greater than those of the remaining
samples.
Stirred-type yogurts
Stirred yogurt from cow milk with The combination of Compared to the hydrodynamic cavitation The pressure homogenization of the milk Gregersen et al. [119]
different fat homogenization CARS and 2 PE process, pressure homogenization resulted base produced a substantially stronger gel
levels and denaturation of whey in a greater degree of milk fat globule structure (higher G′ value) than the
protein (MFG) integration into the protein hydrodynamic cavitation process when
network. milk was not exposed to an extra heat
treatment.
Low-fat, stirred yogurt with CLSM The substitution of fat with MWP resulted The apparent viscosity values of some Torres et al. [30]
microparticulated whey protein in a distinct protein network. In general, an yogurt samples increased as the amount of
(MWP) (as fat replacer) increase in MWP concentration produced a additional whey protein microparticles was
more compact structure composed of big increased. Also, it helped to consist of a
aggregates. stronger gel structure depending on the
kind of microparticulate component used.
Fat-free, stirred yogurts with CLSM Whey protein (WP) addition to yogurt The yogurts produced with a high Laiho et al. [97]
different casein to whey protein resulted in forming larger protein clusters, concentration of WP displayed greater
ratio whereas control yogurt showed a more yield stress and a stronger gel structure.
spaced structure.
Stirred yogurt from cow milk with SEM In contrast to microcapsule yogurts, the The storage modulus, apparent viscosity, Li et al. [138]
synbiotic microcapsules control sample exhibited a continuous and complex modulus values of yogurts
branching network with a greater devoid containing microcapsules were found to be
area, casein aggregates and pores, and firm higher than control samples.
texture.
Yogurts produced with the CLSM The control sample was characterized by The application of micro- and nano-bubbles Babu, Liu, and
application of micro- and nano- its heterogeneous microstructure, which dramatically reduced the viscosity of Amamcharla [139]
bubbles consisted of dense gel clusters of protein yogurt samples.
particles. Yogurt samples treated with
micro- and nano-bubbles exhibited less
dense protein aggregates.
Yogurts produced from raw, SEM Comparatively, the UHT yogurt included Viscosity values of yogurt samples Célia et al. [140]
pasteurized, or UHT milk fewer empty areas than the other produced from UHT and pasteurized milk
treatments. were found to be higher than the samples
manufactured with refrigerated raw milk.
Non-fat stirred yogurt with Optical microscopy Separately from the whey protein level, As whey protein content was decreased, Gilbert et al. [71]
different milk smoothing raising the smoothing temperature resulted yogurt stiffness fell as well. Although
temperatures and protein in larger microgels (dense protein firmness increased as smoothing
composition aggregates). temperature increased, flow characteristics
remained unaffected.
(continued on next page)
7
Table 2 (continued )
Yogurt Instrument for Microstructure observation Rheological effect Reference
microstructural
analyses
Non-fat, low-fat, and high-fat SEM Brand 1(fat-containing samples have a The zero-shear viscosity of Brand 1 samples Erturk et al. [109]
yogurt from two commercial cryo-SEM higher amount of fat compared to Brand 2) was found to be higher than that of Brand 2
brands had a casein and whey protein network samples, which is affected by molecular
that was denser and finer than Brand 2, entanglements.
which had a structure that was almost
similar to a honeycomb.
Fat-free yogurt is produced with a TEM Images captured by transmission electron The chilled yogurt samples had higher Trejo, Corzo-Martínez,
cold step during the fermentation microscopy reveal the migration of casein apparent viscosity and solid-like behavior. Wilkinson,
(a reduction to 4 ◦ C) in the proteins away from the casein micelle as a Higginbotham, and
temperature for 30–120 min result of the decrease in temperature of Harte [141]
acidified milk (pH 5.2) during
fermentation. This results in yogurt gels
composed of clusters of polydispersed
casein gel.
Fig. 3. Microscopy methods to observe the microstructure of yogurt [135].
various stages of the eating process. spaces between aggregates [97]. Furthermore, in previous studies, it has
Several tools have been used to assess the tribological properties of been reported that tribological properties and particle size of yogurt
foods. The Mini-Traction Machine, rheometers with customized at samples have a strong correlation with the creaminess results of sensory
tachments, and uniaxial compression apparatus with appropriate ac analyses. On the other hand, the only sensory descriptions that linked
cessories are common devices [146]. In tribology, a Stribeck curve is effectively with particle size were lumpy, grainy, and smooth [97,147].
utilized to determine the behavior of food in the mouth cavity by plot As a result, there was not found no general technique to predict the
ting the coefficient of friction against the sliding speed. This curve sensory properties of stirred yogurts [98].
consists of three regimes: boundary, mixed, and hydrodynamic [146]. Interestingly, in the study of Nguyen, Kravchuk, Bhandari, and
The shape, size, deformability, and rigidity of the particles within the Prakash [148], the friction curves obtained from tribological analyses of
food matrix and the lubricant features of the continuous phase have a yogurt samples presented a different pattern than a regular Stribeck
significant effect on the tribological results [13,98]. Subsequently, all curve. A four-zone model has been proposed to explain the friction
ingredients of yogurt can influence its friction behavior with the behavior of yogurt samples due to their complex internal structure. It
particular impacts determined by the amount of addition, the relation has been reported that three tribological parameters (v3: entrainment
ship of the ingredients among them in the yogurt matrix, and the asso speed at the transition point between zones 3 and 4; s2: the slope of zone
ciation of ingredients with slipping surfaces [144]. 2; s4: the slope of zone 4) present a correlation with sensory character
In the study of Laiho et al. [97], different casein-to-whey protein istics, such as smoothness-lumpiness (v3), stickiness and oily coating
ratio is used to produce fat-free yogurts. According to the tribological (s2), and thickness (s4). According to their results, skim (0.1% fat) yogurt
results, the predominant factor in the lubrication of control yogurt was samples had a substantially higher friction coefficient than full-fat (3.8%
the stable, uniform protein network generated by whey proteins linked fat) samples. Furthermore, full-fat (3.8%) yogurt samples showed better
to casein micelles. Nevertheless, the serum phase of the yogurt exerted lubrication features at 35 ◦ C compared to samples with various fat ratios
an important effect on friction as the percentage of whey protein was (0.1% and 1.3%), and gelatin was considered the best hydrocolloid to
increased and the protein network structure altered to have larger improve lubrication properties of skim yogurt rather than xanthan gum,
8
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International Journal of Biological Macromolecules 263 (2024) 130428

Contents lists available at ScienceDirect

International Journal of Biological Macromolecules

Perspectives on the yogurt rheology

1. Introduction through the implementation of several strategies: increasing the milk

Fig. 1. Variable factors contributing to yogurt rheology (created with BioRender).

Fig. 3. Microscopy methods to observe the microstructure of yogurt [135].

carrageenan, and modified starch. The difference in the friction References

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