Functional Dairy Food Products

**81**

**Chapter 6**

**Abstract**

**1. Introduction**

Development of

Functional Cheeses with

*Diana Palatnik, Noelia Rinaldoni, Diego Corrales,* 

this development could represent an innovation for dairy industry.

**Keywords:** functional cheese, fructans, microfiltration membranes

favorable amino acid profile and good digestibility.

Cheeses are high nutritional value foods and with great market demand, outstanding among other products as it has high biological value proteins with a very

Argentine is the seventh largest producer of cheese in the world in a relatively stable market, where consumption per capita is 12 kilos per year. It is reported that half of the country's milk is destined for cheese processing. It is about 500 thousand tons and is distributed 50% to make soft cheeses, 35% for semihard, and 15% for hard. Between 70 and 75% of the production is commercialized in the domestic market: approximately 13,000 dairies supply about 870 establishments that make cheeses . In Latin America, Argentina is the country with the highest consumption of cheeses. However, cheese has restrictions on its consumption due to its high content of calories and highly saturated fats and therefore with a potential risk to health.

*María L. Rolon, Haydée Montero, Germán Aranibar,* 

*María L. Castells, Noemi Zaritzky and Mercedes E. Campderrós*

Cheese is a food of great consumption in the world; however, some aspects related to its fat content and the possibility of incorporating fiber represent interesting challenges for the dairy industry. In this sense, fructooligosaccharides (FOS), as inulin and agave fructans, exhibit valuable nutritional and functional attributes that can be used as supplements as soluble fiber or as macronutrient substitutes. In this chapter, the study of the development of soft and cream cheeses was performed to determine the operating conditions that allow obtaining products with beneficial health properties taking advantage of the characteristics of this carbohydrate. The skim milk was produced by ultrafiltration, and all the products were characterized physicochemically, including determinations of color, texture, and sensory analysis. The cheeses obtained were of high moisture, >45% (w/w), and reduced fat content (10–25% w/w), including a high protein concentration. The presence of fructans did not significantly modify the texture and appearance of the developed products, but its retention in the matrix was maximal in the case of spreadable cream cheeses containing inulin. Considering the health benefits of fructans and their abundance,

Fructooligosaccharides

#### **Chapter 6**

## Development of Functional Cheeses with Fructooligosaccharides

*Diana Palatnik, Noelia Rinaldoni, Diego Corrales, María L. Rolon, Haydée Montero, Germán Aranibar, María L. Castells, Noemi Zaritzky and Mercedes E. Campderrós*

#### **Abstract**

Cheese is a food of great consumption in the world; however, some aspects related to its fat content and the possibility of incorporating fiber represent interesting challenges for the dairy industry. In this sense, fructooligosaccharides (FOS), as inulin and agave fructans, exhibit valuable nutritional and functional attributes that can be used as supplements as soluble fiber or as macronutrient substitutes. In this chapter, the study of the development of soft and cream cheeses was performed to determine the operating conditions that allow obtaining products with beneficial health properties taking advantage of the characteristics of this carbohydrate. The skim milk was produced by ultrafiltration, and all the products were characterized physicochemically, including determinations of color, texture, and sensory analysis. The cheeses obtained were of high moisture, >45% (w/w), and reduced fat content (10–25% w/w), including a high protein concentration. The presence of fructans did not significantly modify the texture and appearance of the developed products, but its retention in the matrix was maximal in the case of spreadable cream cheeses containing inulin. Considering the health benefits of fructans and their abundance, this development could represent an innovation for dairy industry.

**Keywords:** functional cheese, fructans, microfiltration membranes

#### **1. Introduction**

Cheeses are high nutritional value foods and with great market demand, outstanding among other products as it has high biological value proteins with a very favorable amino acid profile and good digestibility.

Argentine is the seventh largest producer of cheese in the world in a relatively stable market, where consumption per capita is 12 kilos per year. It is reported that half of the country's milk is destined for cheese processing. It is about 500 thousand tons and is distributed 50% to make soft cheeses, 35% for semihard, and 15% for hard. Between 70 and 75% of the production is commercialized in the domestic market: approximately 13,000 dairies supply about 870 establishments that make cheeses . In Latin America, Argentina is the country with the highest consumption of cheeses.

However, cheese has restrictions on its consumption due to its high content of calories and highly saturated fats and therefore with a potential risk to health.

Indeed, one of the market trends is the production of cheeses with reduced fat content to minimize the negative effects of lipids in the diet. With the purpose of texturally compensating the product, the addition of fructans such as inulin or agave fructans has been studied [1, 2].

Fructans are nondigestible carbohydrates present in many vegetables, fruits, and cereals. They are widely used as ingredients of functional food since they have beneficial effects on health as prebiotics. Dietary prebiotics are typically nondigestible fiber compounds that pass undigested through the upper part of the gastrointestinal tract and stimulate the growth or activity of advantageous bacteria that colonize the large bowel by acting as substrate for them. They supply dietary fiber, hypoglycemic, low calorie value, providing a better bioavailability of calcium and magnesium and improving intestinal iron absorption [3, 4]. Bosscher et al. [5] reported that the consumption of fructans in humans increases calcium absorption, improving bone mineral content and density. Besides, there are promising evidences of its performance in the regulation of lipid parameter, reduction of cancer risk, reinforcement of the immune response, and protection against bowel disorders.

According to the American Dietetic Association (ADA), it is usually understood under the name of functional food any food or potentially healthy ingredient that can provide health benefits beyond the traditional nutrients it contains.

Hippócrates, a Greek doctor more than 2000 years ago, left in his legacy a mythical phrase "That food be your medicine and medicine your food" and, although he did not use the term functional food, was implicitly referring to the consumption of certain foods that could help prevent diseases. Located in the twenty-first century, this philosophy of "food as medicine" is the basis of the paradigm of functional foods. Regarding the abovementioned, the challenge of achieving functional consumer cheeses is undoubtedly an interesting aspect for the dairy industry.

The basic technology of manufacturing a cheese is similar for almost all varieties, but it is important to mention that relatively small changes in the processing conditions give rise to important differences in the final cheese. In general these differences lie in the use of different types of ferments, in variations in cooking temperatures, curd cut, cheese grain size, brine and ripening times, and other technological conditions. Furthermore, in the case that different ingredients that provide the desired functionality are used, it also leads to really different products.

The membrane technology is increasingly employed for dairy treatment [6]. The development of membrane materials, equipment, and studies of fouling and cleaning of membranes increases their applications. These aspects are addressed in this chapter with the purpose of achieving a functional food from natural resources also using the membrane technology to perform the skimming of the raw material.

In this chapter, results about the incorporation of fructans in cheesemaking matrices are described in order to obtain reduced fat cheese-containing compounds that behave like probiotics and act as dietary fiber.

#### **2. Materials and methods**

#### **2.1 Raw materials**

The elaboration of each of the samples of cheese studied was made from 2500 ml of raw milk from a dairy farm in the province of San Luis and with milk sample of an establishment of the province of Buenos Aires. Before starting the cheese manufacturing process, a fat reduction was carried out in order to obtain a product that is low in fat and, therefore, healthier. For this purpose, a procedure using membrane technology was developed, as will be explained later.

**83**

technology.

*Development of Functional Cheeses with Fructooligosaccharides*

The following compounds were added to all cheese samples per liter of milk: ferment consisting of lactic acid bacteria that allow the acidification and coagulation of milk, inhibiting, in addition, the development of other unwanted microorganisms. There are different types of ferments, and the one chosen depends on the type of cheese. In this work homofermentative mesophyll ferment, lyophilized direct inoculation composed of *Lactococcus lactis* subsp. *lactis* and *Lactococcus lactis* subsp. *cremoris* (CHR Hansen R-704), was used. Although the ferment is a direct inoculation, it is convenient to leave it pre- mature for 30 min in a little amount of milk at an optimum temperature of 34 ± 2°C before adding it to the rest of the milk. The working temperature was also maintained at 32–37°C during a large part of the processing to allow the action of the ferment, rennet (CHR Hansen, Chy Max M200–0.5 ml). FOS of different characteristics were used: (i) inulin provided by Orafti Chile SA and obtained from chicory and (ii) agave native fructans (NF), commercial products of *Agave tequilana* in powder and 72°Brix (Agavetica Monterrey, Mexico). The commercial inulin employed is mainly constituted by linear chains of fructose, with a glucose terminal unit. Short-chain inulin GR was used in concentrations of 3 and 5% (w/w) and long-chain HP at 5% (w/v). Fructans present in Agave, particularly in *Agave tequilana*, have a polymerization degree between 3 and 29 with several β

The amount of fructan added was defined taking into account the conditions set

For milk skimming, membrane technology was used, more precisely the microfiltration technology. Once skimmed, a physicochemical analysis was carried out,

As mentioned previously, current market demands healthier foods with lower health risks. Thus, it is expected that in a cheese with functional properties, its fat content will be reduced [1]. For the elaboration of the samples of cheeses, raw cow's milk was used, and the fat content was reduced by applying membrane

Membrane technology is a generic term for a series of different and very distinctive separation processes. In essence, the membrane acts as a selective barrier by letting some components pass through it and retaining others depending on the molecular size, with a pressure gradient being the driving force of the process. The feed (crude milk) was pumped through a frontal polyethylene microfiltration (MF) filter with a pore size of 5–10 μm (Pall Corporation, USA) at room temperature (24 ± 1°C), which is shown in **Figure 1**. Applied transmembrane pressure and recirculation rate were controlled by a valve and by the feed rate, supplied by the variable speed pump from a feed tank. The pressure applied to the membrane was measured with a manometer, and the ermeate was determined by measuring the filtrate volume collected during a certain period of time. This procedure reduced the amount of bacteria and spores and acted as cold pasteurization. The filter was cleaned and sanitized after each experiment and exchanged periodically. The operation was carried out in three stages, taking a sample from each of them, in order to determine the degree of skimming of the milk in each step for

Partially skimmed milk was divided in different portions of 2.5 L for each batch. One of the portions was reserved to produce a low-fat cheese as a control

by the Argentine Food Code (AFC), Chap. 17, for foods added with fibers.

*DOI: http://dx.doi.org/10.5772/intechopen.85888*

(2–6) bonds in branching fructose molecule.

and the process of making the cheeses began.

which the fat content was determined.

**2.3 Soft cheese manufacture**

**2.2 Membrane processes for partial skim milk**

*Development of Functional Cheeses with Fructooligosaccharides DOI: http://dx.doi.org/10.5772/intechopen.85888*

*Current Issues and Challenges in the Dairy Industry*

agave fructans has been studied [1, 2].

Indeed, one of the market trends is the production of cheeses with reduced fat content to minimize the negative effects of lipids in the diet. With the purpose of texturally compensating the product, the addition of fructans such as inulin or

Fructans are nondigestible carbohydrates present in many vegetables, fruits, and cereals. They are widely used as ingredients of functional food since they have beneficial effects on health as prebiotics. Dietary prebiotics are typically nondigestible fiber compounds that pass undigested through the upper part of the gastrointestinal tract and stimulate the growth or activity of advantageous bacteria that colonize the large bowel by acting as substrate for them. They supply dietary fiber, hypoglycemic, low calorie value, providing a better bioavailability of calcium and magnesium and improving intestinal iron absorption [3, 4]. Bosscher et al. [5] reported that the consumption of fructans in humans increases calcium absorption, improving bone mineral content and density. Besides, there are promising evidences of its performance in the regulation of lipid parameter, reduction of cancer risk, reinforcement

According to the American Dietetic Association (ADA), it is usually understood under the name of functional food any food or potentially healthy ingredient that

Hippócrates, a Greek doctor more than 2000 years ago, left in his legacy a mythical phrase "That food be your medicine and medicine your food" and, although he did not use the term functional food, was implicitly referring to the consumption of certain foods that could help prevent diseases. Located in the twenty-first century, this philosophy of "food as medicine" is the basis of the paradigm of functional foods. Regarding the abovementioned, the challenge of achieving functional consumer cheeses is undoubtedly an interesting aspect for the dairy industry.

The basic technology of manufacturing a cheese is similar for almost all varieties, but it is important to mention that relatively small changes in the processing conditions give rise to important differences in the final cheese. In general these differences lie in the use of different types of ferments, in variations in cooking temperatures, curd cut, cheese grain size, brine and ripening times, and other technological conditions. Furthermore, in the case that different ingredients that provide the desired functionality are used, it also leads to really different products. The membrane technology is increasingly employed for dairy treatment [6]. The development of membrane materials, equipment, and studies of fouling and cleaning of membranes increases their applications. These aspects are addressed in this chapter with the purpose of achieving a functional food from natural resources also using the membrane technology to perform the skimming of the raw material. In this chapter, results about the incorporation of fructans in cheesemaking matrices are described in order to obtain reduced fat cheese-containing compounds

The elaboration of each of the samples of cheese studied was made from 2500 ml of raw milk from a dairy farm in the province of San Luis and with milk sample of an establishment of the province of Buenos Aires. Before starting the cheese manufacturing process, a fat reduction was carried out in order to obtain a product that is low in fat and, therefore, healthier. For this purpose, a procedure using membrane

of the immune response, and protection against bowel disorders.

that behave like probiotics and act as dietary fiber.

technology was developed, as will be explained later.

**2. Materials and methods**

**2.1 Raw materials**

can provide health benefits beyond the traditional nutrients it contains.

**82**

The following compounds were added to all cheese samples per liter of milk: ferment consisting of lactic acid bacteria that allow the acidification and coagulation of milk, inhibiting, in addition, the development of other unwanted microorganisms. There are different types of ferments, and the one chosen depends on the type of cheese. In this work homofermentative mesophyll ferment, lyophilized direct inoculation composed of *Lactococcus lactis* subsp. *lactis* and *Lactococcus lactis* subsp. *cremoris* (CHR Hansen R-704), was used. Although the ferment is a direct inoculation, it is convenient to leave it pre- mature for 30 min in a little amount of milk at an optimum temperature of 34 ± 2°C before adding it to the rest of the milk. The working temperature was also maintained at 32–37°C during a large part of the processing to allow the action of the ferment, rennet (CHR Hansen, Chy Max M200–0.5 ml).

FOS of different characteristics were used: (i) inulin provided by Orafti Chile SA and obtained from chicory and (ii) agave native fructans (NF), commercial products of *Agave tequilana* in powder and 72°Brix (Agavetica Monterrey, Mexico). The commercial inulin employed is mainly constituted by linear chains of fructose, with a glucose terminal unit. Short-chain inulin GR was used in concentrations of 3 and 5% (w/w) and long-chain HP at 5% (w/v). Fructans present in Agave, particularly in *Agave tequilana*, have a polymerization degree between 3 and 29 with several β (2–6) bonds in branching fructose molecule.

The amount of fructan added was defined taking into account the conditions set by the Argentine Food Code (AFC), Chap. 17, for foods added with fibers.

For milk skimming, membrane technology was used, more precisely the microfiltration technology. Once skimmed, a physicochemical analysis was carried out, and the process of making the cheeses began.

#### **2.2 Membrane processes for partial skim milk**

As mentioned previously, current market demands healthier foods with lower health risks. Thus, it is expected that in a cheese with functional properties, its fat content will be reduced [1]. For the elaboration of the samples of cheeses, raw cow's milk was used, and the fat content was reduced by applying membrane technology.

Membrane technology is a generic term for a series of different and very distinctive separation processes. In essence, the membrane acts as a selective barrier by letting some components pass through it and retaining others depending on the molecular size, with a pressure gradient being the driving force of the process.

The feed (crude milk) was pumped through a frontal polyethylene microfiltration (MF) filter with a pore size of 5–10 μm (Pall Corporation, USA) at room temperature (24 ± 1°C), which is shown in **Figure 1**. Applied transmembrane pressure and recirculation rate were controlled by a valve and by the feed rate, supplied by the variable speed pump from a feed tank. The pressure applied to the membrane was measured with a manometer, and the ermeate was determined by measuring the filtrate volume collected during a certain period of time. This procedure reduced the amount of bacteria and spores and acted as cold pasteurization. The filter was cleaned and sanitized after each experiment and exchanged periodically.

The operation was carried out in three stages, taking a sample from each of them, in order to determine the degree of skimming of the milk in each step for which the fat content was determined.

#### **2.3 Soft cheese manufacture**

Partially skimmed milk was divided in different portions of 2.5 L for each batch. One of the portions was reserved to produce a low-fat cheese as a control

**Figure 1.** *Frontal MF equipment, used in the skimming of raw milk. (T = 24 ± 1°C): 1: feed tank; 2: pump; 3: MF filter.*

(LFCC) sample, and to the rest of the portions, different fructans as inulin GR and HP (at 3 and 5% w/v) and NF [in concentrations of 0.5 and 5% (w/v)] were added. Full-fat control cheeses (FFCC), without fructans, were also produced for comparison.

Different tests were carried out in the stage of FOS aggregation, searching the way in which it would be better retained within the cheese matrix and thus obtaining a food rich in fibers. The alternatives assayed in cheeses elaboration are described as follows:

a.Preparation of samples with added inulin powder

All samples were made in duplicate. The inulin powder previously weighed according to the desired final percentage was added. The addition was done smoothly and slowly by stirring continuously to avoid the formation of lumps. Once the addition of the carbohydrate was done, the elaboration was continued according to the steps described in **Figure 2**.

The samples obtained by this process were labeled as M1 and M2:


Other forms of addition were investigated, such as the addition of carbohydrate prior to the formation of a gel [7]. Inulin shows high-level gelling properties, forming a three-dimensional network of insoluble submicron particles with a large amount of immobilized water, which ensures physical stability.

Regarding the gels made in the laboratory, it was found that the gel with 30% inulin with a heating time of 20 min was the one that exhibited the best consistency. In this way, the cheese samples were developed with incorporated inulin as gel (formed at 30% w/v) in different stages of the elaboration, resulting in samples M3 and M4 (**Figure 2**):

**85**

**Figure 2.**

*Development of Functional Cheeses with Fructooligosaccharides*

• M3: Addition after the incorporation of the rennet and before the curd is formed

• M4: Incorporation of the gel after the syneresis stage, mixing, and molding

NF in concentrations of 0.5% and 5% (w/v) was added in powder in the same

Production was carried out in a batch process. Each formulation was replicated at least twice and analyzed independently; also each set of samples was made in the

c.Preparation of samples with added native agave fructans powder

way as inulin, as is indicated in **Figure 2**.

*Flowchart of the soft cheese making process with and without FOS.*

The samples obtained were identified as:

same day. The initial pH was measured (6.89 ± 0.10).

• M5: addition of FN 0.5%

• M6: addition of FN 5%

*DOI: http://dx.doi.org/10.5772/intechopen.85888*

*Development of Functional Cheeses with Fructooligosaccharides DOI: http://dx.doi.org/10.5772/intechopen.85888*

#### **Figure 2.**

*Current Issues and Challenges in the Dairy Industry*

(LFCC) sample, and to the rest of the portions, different fructans as inulin GR and HP (at 3 and 5% w/v) and NF [in concentrations of 0.5 and 5% (w/v)] were added. Full-fat control cheeses (FFCC), without fructans, were also produced for

*Frontal MF equipment, used in the skimming of raw milk. (T = 24 ± 1°C): 1: feed tank; 2: pump; 3: MF filter.*

Different tests were carried out in the stage of FOS aggregation, searching the way in which it would be better retained within the cheese matrix and thus obtaining a food rich in fibers. The alternatives assayed in cheeses elaboration are

All samples were made in duplicate. The inulin powder previously weighed according to the desired final percentage was added. The addition was done smoothly and slowly by stirring continuously to avoid the formation of lumps. Once the addition of the carbohydrate was done, the elaboration was contin-

Other forms of addition were investigated, such as the addition of carbohydrate prior to the formation of a gel [7]. Inulin shows high-level gelling properties, forming a three-dimensional network of insoluble submicron particles with a large amount of immobilized water, which ensures physical

Regarding the gels made in the laboratory, it was found that the gel with 30% inulin with a heating time of 20 min was the one that exhibited the best consistency. In this way, the cheese samples were developed with incorporated inulin as gel (formed at 30% w/v) in different stages of the elaboration, result-

The samples obtained by this process were labeled as M1 and M2:

a.Preparation of samples with added inulin powder

ued according to the steps described in **Figure 2**.

• M1: sample with 125 g of inulin powder (5% w/v)

b.Preparation of samples with added inulin in gel

ing in samples M3 and M4 (**Figure 2**):

• M2: sample with 37.5 g of inulin powder (1.5% w/v)

**84**

stability.

comparison.

**Figure 1.**

described as follows:

*Flowchart of the soft cheese making process with and without FOS.*


NF in concentrations of 0.5% and 5% (w/v) was added in powder in the same way as inulin, as is indicated in **Figure 2**.

The samples obtained were identified as:


Production was carried out in a batch process. Each formulation was replicated at least twice and analyzed independently; also each set of samples was made in the same day. The initial pH was measured (6.89 ± 0.10).

Pasteurization is an operation that is carried out to destroy the pathogenic flora and reduce the banal flora or alteration, so the milk was heated to 65 ± 1°C and kept at this temperature for 30 min. After that time it was quickly cooled using an ice bath. After the addition of fructans in any way, the samples were mixed. Then the temperature was brought to 34.5 ± 2°C, and 1 g of CaCl2 (BDH Chemicals Laboratory Reagents, United Kingdom) was added which contributes to obtain a proper floc. Then the ferment (0.1 g) was added to the batches. When the pH was at 6.5 ± 0.2, the temperature was raised to 37.5 ± 0.5°C, and enzymatic coagulant in powder (0.1 g) was added for coagulation, allowing acting for 40 min. To make sure that the coagulation has come to an end, a knife was placed in the dough, and it was checked that it comes out dry, without curd remains, which indicates that it has already been formed.

Cutting was done 35 ± 5 min later, the grains remained in suspension, and the dough was heated 2°C above the coagulation temperature and was gently stirred (avoiding breaking the curd cubes) and allowed to settle for 10 min. The curd was cut with a curd knife in the shape of 2-cm3 cubes. The cut curd was allowed to settle for 10 min. Then the temperature was increased by 2 ± 0.5°C while gently mixing so as not to break the curd granules. After that, the samples were poured onto a sieve covered with a cheesecloth for drainage of whey, which was collected in a graduated cylinder. This process facilitates the release of the grain serum from the dough. The cheese curds were put into cylindrical plastic molds of 250 g (Vigna S.A., Argentina) and pressed during 1 h and half of each side (**Figure 3a**). A posterior mold-pressing stage was carried out in an artisanal cheese press (**Figure 3b**) and adjusted to exert pressure on the molds allowing the draining of excess serum. The pressing time was varying between the different batches according to the pH which was decreased to the desired value: 5.4 ± 0.1. Then the samples were placed in saturated brine, calculating the dwell time in the brine according to the weight of the cheeses. The salt regulates the development of microorganisms, enhancing flavors and contributing to the formation of the product's crust. The brine was prepared at a concentration of 21°Baumé (°Be), which is the recommended concentration for soft cheese.

The samples were remained at 6 ± 2°C in a refrigerator and after 24 h were packaged with a plastic film.

#### **2.4 Preadable cream cheese manufacture with added inulin**

The addition of native or short-chain inulin (GR) and high-performance (HP) inulin in cream cheeses was studied. These samples were elaborated in the National

**87**

**Figure 4.**

*Development of Functional Cheeses with Fructooligosaccharides*

Institute of Industrial Technology (INTI of Argentina). **Figure 4** shows the general flow diagram used. In this case the raw material (milk) was provided by Mastellone

The elaboration process was in batch of 20 l of milk. Once the milk was received, a filtering was done to eliminate impurities, and physicochemical determinations

Pasteurization was carried out at 85 ± 1°C for 30 min. As in the soft cheeses with inulin, after pasteurization, the milk was quickly cooled in an ice bath until reaching

Cooling

Filtered out

Milk reception

Pasteurization 85 °C-30'

Additive additives

Cl2Ca Ferment Coagulant

Curd formation 32° until pH 5.4

> Cuttimg 4x4 cm

 Incubation until pH 4.8

Filtered in cold room

Whey drainage

Kneading

Packing and Storage 2-6ºC

*DOI: http://dx.doi.org/10.5772/intechopen.85888*

S.A. (Buenos Aires, Argentine).

Inulin HP **(M7)**

Inulin GR **(M8)**

*Flowchart of the cream cheese making process.*

were performed.

**Figure 3.** *(a) Plastic molds with cheese samples, (b) cheese press.*

*Current Issues and Challenges in the Dairy Industry*

cut with a curd knife in the shape of 2-cm3

aged with a plastic film.

Pasteurization is an operation that is carried out to destroy the pathogenic flora and reduce the banal flora or alteration, so the milk was heated to 65 ± 1°C and kept at this temperature for 30 min. After that time it was quickly cooled using an ice bath. After the addition of fructans in any way, the samples were mixed. Then the temperature was brought to 34.5 ± 2°C, and 1 g of CaCl2 (BDH Chemicals Laboratory Reagents, United Kingdom) was added which contributes to obtain a proper floc. Then the ferment (0.1 g) was added to the batches. When the pH was at 6.5 ± 0.2, the temperature was raised to 37.5 ± 0.5°C, and enzymatic coagulant in powder (0.1 g) was added for coagulation, allowing acting for 40 min. To make sure that the coagulation has come to an end, a knife was placed in the dough, and it was checked that it comes out dry, without curd remains, which indicates that it has already been formed. Cutting was done 35 ± 5 min later, the grains remained in suspension, and the dough was heated 2°C above the coagulation temperature and was gently stirred (avoiding breaking the curd cubes) and allowed to settle for 10 min. The curd was

for 10 min. Then the temperature was increased by 2 ± 0.5°C while gently mixing so as not to break the curd granules. After that, the samples were poured onto a sieve covered with a cheesecloth for drainage of whey, which was collected in a graduated cylinder. This process facilitates the release of the grain serum from the dough. The cheese curds were put into cylindrical plastic molds of 250 g (Vigna S.A., Argentina) and pressed during 1 h and half of each side (**Figure 3a**). A posterior mold-pressing stage was carried out in an artisanal cheese press (**Figure 3b**) and adjusted to exert pressure on the molds allowing the draining of excess serum. The pressing time was varying between the different batches according to the pH which was decreased to the desired value: 5.4 ± 0.1. Then the samples were placed in saturated brine, calculating the dwell time in the brine according to the weight of the cheeses. The salt regulates the development of microorganisms, enhancing flavors and contributing to the formation of the product's crust. The brine was prepared at a concentration of

21°Baumé (°Be), which is the recommended concentration for soft cheese.

**2.4 Preadable cream cheese manufacture with added inulin**

The samples were remained at 6 ± 2°C in a refrigerator and after 24 h were pack-

The addition of native or short-chain inulin (GR) and high-performance (HP) inulin in cream cheeses was studied. These samples were elaborated in the National

cubes. The cut curd was allowed to settle

**86**

**Figure 3.**

*(a) Plastic molds with cheese samples, (b) cheese press.*

Institute of Industrial Technology (INTI of Argentina). **Figure 4** shows the general flow diagram used. In this case the raw material (milk) was provided by Mastellone S.A. (Buenos Aires, Argentine).

The elaboration process was in batch of 20 l of milk. Once the milk was received, a filtering was done to eliminate impurities, and physicochemical determinations were performed.

Pasteurization was carried out at 85 ± 1°C for 30 min. As in the soft cheeses with inulin, after pasteurization, the milk was quickly cooled in an ice bath until reaching

**Figure 4.** *Flowchart of the cream cheese making process.*

**Figure 5.** *(a) Kneader used in the production of cream cheese (b) and cream cheese.*

33 ± 1°C, optimum temperature for the addition of ferment and other additives. In this case, 0.042 g/l of ferment (Sacco M032), 0.13 g/l of CaCl2 (78% purity), and 0.5 ml/l of rennet Chy Max M200 (1/100 dilution) were added, and it was left incubating until it reached pH 5.4. A control sample without inulin was reserved, and the rest incorporated the polysaccharide in different percentages. Short-chain inulin GR was used in concentrations of 3 and 5% (w/w) and long-chain HP at 5% (w/v).

Once the curd was formed, cuts of 4 × 4 cm were made with a lyre and led to take out the whey in a cold chamber until reaching a pH of 4.8 ± 0.1; for this, a cheesecloth was used, like in the production of soft cheeses. It was allowed to run through the night, and then the shaking or texturing stage was reached with a Hobart mixer (USA), shown in **Figure 5a**. The process consisted of 30 s of shaking at speed 1 and then 1.5 min at speed 2, adding inulin during this stage. After shaking, the product was placed in plastic pots (**Figure 5b**) to be taken to the cold room where they were stored for 24 h before making the physicochemical determinations.

Inulin GR (native or short-chain inulin) was added in the texturing or kneading stage, in the form of rain, continuously kneading until a homogenous mixture was obtained. The amounts added were 3% (28.35 g) and 5% (47.25 g), obtaining the samples denominated as M7, M8. HP inulin was added in the pasteurization stage, since it is much more soluble than GR and only in the 5% concentration (47.25 g) reaching to sample M9.

#### **2.5 Analysis**

Raw materials and cheeses samples were analyzed in duplicate according to standard replication AOAC [8]. Analyses were performed after 48 h of sample elaboration.

pH was measured using a digital pH-meter (Testo 206-pH 2, Germany).

The protein content was calculated by determination of total nitrogen by the Kjeldahl method using a Digestion Blocks and a semiautomatic Kjeldahl distiller (Selecta, Spain); the conversion factor used to express the results was 6.38 (AOAC 991.22). The fat content was measured by the Rosse-Gottlieb method (AOAC 933.05). Total solids were determined by weight difference, drying in an oven at 70 ± 1°C, during 24 h (AOAC 925.23). The moisture content was determined by

**89**

*Development of Functional Cheeses with Fructooligosaccharides*

temperature programmer to reach 520°C (AOAC 945.46).

The assessment was performed in triplicate at 9 ± 2°C.

analysis of variance using the statistical software GraphPad InStat.

fromages a pâte dure ou semidure," 1994, of the European Union.

20,001/20,002/20,004/20,005 and 20,006.

The analysis of samples of cream cheese were carried up by a panel composed of 10 blind and visually impaired people belonging to INTI-Dairy (Buenos Aires), which was selected and trained according to IRAM Norms

For the determination of the texture and flavor profiles, the Quantitative Descriptive Analysis (QDA) technique contemplated in IRAM Standards 20,012 and 20,013 was used. These standards contemplate the analysis of the texture profile and procedures indicated in the "Guide D'Evaluation Sensorielle de la Texture des

From the texture profile, the following descriptors were determined: (a) adherence, work that must be done with the tongue to detach a product stuck on the palate and the teeth; (b) solubility, a sensation that is highlighted when the sample melts very quickly in the saliva after chewing it four times with the

gravimetric method (IDF 1982). For ash determination, samples were weighted into porcelain crucibles and incinerated in a muffle furnace (Indef, Argentina) with a

The determination of total carbohydrates (lactose plus fructan) was carried out in whey, sub-product of cheese manufacture, using a refractometer (Arcano, China, range 0–32°Brix) in which the soluble compounds are expressed as °Brix. The measurement takes only few seconds [2]. The determination of fructan was made by difference between the amount of total carbohydrates measured and the lactose recorded in the control sample. The fructan value retained in cheeses was obtained by difference between the amount of fructan added and the amount found in the whey. Also inulin was determined by anion-exchange high-performance liquid chromatography (HPLC) method following water extraction of inulin. HPLC conditions included an Aminex HPX-87C column (Bio-Rad), deionized water at 85°C as the mobile phase with a flow rate of 0.6 ml/min, and a refractive index detector [9]. The surface color was measured by a digital spectrophotometer (Mini Scan EZ, USA) provided with the software. The chromometer was calibrated with the standard white and black color. The results reported are averages of measurements of three slices (five measurements per slice), using CIELAB L\*, a\*, and b\* values. L\* value is the lightness variable from 100 for perfect white to zero for black, while a\* and b\* values are the chromaticity values, +redness/-greenness and + yellowness/-blueness, respectively. Instrumental texture analysis was determined using the TAXT Stable

Microsystems analyzer (London, United Kingdom). A compression test was carried out, using a 5-mm cylindrical test tube and with the following test parameters: pretest speed, 2 mm/seg; test speed, 1 mm/seg; distance, 10 mm; and trigger, 3 g.

The sensory evaluation was carried out following different protocols. The samples of soft cheese were tested in a uniformly illuminated room, by 45 members of a semi-trained panel selected from a pool of students and staff members of our department. Prior to assessment, each model cheese sample was divided into various portions and equilibrated at room temperature (22 ± 2°C). A discrimination test was employed in which the evaluator had to establish the difference between a control sample and one or more problem samples, using a scale from 0 (no difference) to 6 (very much different). The samples were coded with three-digit random numbers and were presented in pairs: control vs. sample and control vs. control (as blind witness). The attributes evaluated comparatively were flavor, color, texture, sweetness, and acidity. Panelists were exposed to each sample on an individual Petri plastic dish and were asked to assess a number of specific attributes. Water was provided for rinsing between samples, to clean the palate [10]. The average for all attributes of the sample was calculated, and the differences were analyzed with the

*DOI: http://dx.doi.org/10.5772/intechopen.85888*

#### *Development of Functional Cheeses with Fructooligosaccharides DOI: http://dx.doi.org/10.5772/intechopen.85888*

*Current Issues and Challenges in the Dairy Industry*

33 ± 1°C, optimum temperature for the addition of ferment and other additives. In this case, 0.042 g/l of ferment (Sacco M032), 0.13 g/l of CaCl2 (78% purity), and 0.5 ml/l of rennet Chy Max M200 (1/100 dilution) were added, and it was left incubating until it reached pH 5.4. A control sample without inulin was reserved, and the rest incorporated the polysaccharide in different percentages. Short-chain inulin GR was used in concentrations of 3 and 5% (w/w) and long-chain HP at 5% (w/v). Once the curd was formed, cuts of 4 × 4 cm were made with a lyre and led to take out the whey in a cold chamber until reaching a pH of 4.8 ± 0.1; for this, a cheesecloth was used, like in the production of soft cheeses. It was allowed to run through the night, and then the shaking or texturing stage was reached with a Hobart mixer (USA), shown in **Figure 5a**. The process consisted of 30 s of shaking at speed 1 and then 1.5 min at speed 2, adding inulin during this stage. After shaking, the product was placed in plastic pots (**Figure 5b**) to be taken to the cold room where they were stored for 24 h before making the physicochemical determinations. Inulin GR (native or short-chain inulin) was added in the texturing or kneading stage, in the form of rain, continuously kneading until a homogenous mixture was obtained. The amounts added were 3% (28.35 g) and 5% (47.25 g), obtaining the samples denominated as M7, M8. HP inulin was added in the pasteurization stage, since it is much more soluble than GR and only in the 5% concentration (47.25 g)

*(a) Kneader used in the production of cream cheese (b) and cream cheese.*

**(a) (b)** 

Raw materials and cheeses samples were analyzed in duplicate according to standard replication AOAC [8]. Analyses were performed after 48 h of sample

pH was measured using a digital pH-meter (Testo 206-pH 2, Germany). The protein content was calculated by determination of total nitrogen by the Kjeldahl method using a Digestion Blocks and a semiautomatic Kjeldahl distiller (Selecta, Spain); the conversion factor used to express the results was 6.38 (AOAC 991.22). The fat content was measured by the Rosse-Gottlieb method (AOAC 933.05). Total solids were determined by weight difference, drying in an oven at 70 ± 1°C, during 24 h (AOAC 925.23). The moisture content was determined by

**88**

reaching to sample M9.

**2.5 Analysis**

**Figure 5.**

elaboration.

gravimetric method (IDF 1982). For ash determination, samples were weighted into porcelain crucibles and incinerated in a muffle furnace (Indef, Argentina) with a temperature programmer to reach 520°C (AOAC 945.46).

The determination of total carbohydrates (lactose plus fructan) was carried out in whey, sub-product of cheese manufacture, using a refractometer (Arcano, China, range 0–32°Brix) in which the soluble compounds are expressed as °Brix. The measurement takes only few seconds [2]. The determination of fructan was made by difference between the amount of total carbohydrates measured and the lactose recorded in the control sample. The fructan value retained in cheeses was obtained by difference between the amount of fructan added and the amount found in the whey.

Also inulin was determined by anion-exchange high-performance liquid chromatography (HPLC) method following water extraction of inulin. HPLC conditions included an Aminex HPX-87C column (Bio-Rad), deionized water at 85°C as the mobile phase with a flow rate of 0.6 ml/min, and a refractive index detector [9].

The surface color was measured by a digital spectrophotometer (Mini Scan EZ, USA) provided with the software. The chromometer was calibrated with the standard white and black color. The results reported are averages of measurements of three slices (five measurements per slice), using CIELAB L\*, a\*, and b\* values. L\* value is the lightness variable from 100 for perfect white to zero for black, while a\* and b\* values are the chromaticity values, +redness/-greenness and + yellowness/-blueness, respectively.

Instrumental texture analysis was determined using the TAXT Stable Microsystems analyzer (London, United Kingdom). A compression test was carried out, using a 5-mm cylindrical test tube and with the following test parameters: pretest speed, 2 mm/seg; test speed, 1 mm/seg; distance, 10 mm; and trigger, 3 g. The assessment was performed in triplicate at 9 ± 2°C.

The sensory evaluation was carried out following different protocols. The samples of soft cheese were tested in a uniformly illuminated room, by 45 members of a semi-trained panel selected from a pool of students and staff members of our department. Prior to assessment, each model cheese sample was divided into various portions and equilibrated at room temperature (22 ± 2°C). A discrimination test was employed in which the evaluator had to establish the difference between a control sample and one or more problem samples, using a scale from 0 (no difference) to 6 (very much different). The samples were coded with three-digit random numbers and were presented in pairs: control vs. sample and control vs. control (as blind witness). The attributes evaluated comparatively were flavor, color, texture, sweetness, and acidity. Panelists were exposed to each sample on an individual Petri plastic dish and were asked to assess a number of specific attributes. Water was provided for rinsing between samples, to clean the palate [10]. The average for all attributes of the sample was calculated, and the differences were analyzed with the analysis of variance using the statistical software GraphPad InStat.

The analysis of samples of cream cheese were carried up by a panel composed of 10 blind and visually impaired people belonging to INTI-Dairy (Buenos Aires), which was selected and trained according to IRAM Norms 20,001/20,002/20,004/20,005 and 20,006.

For the determination of the texture and flavor profiles, the Quantitative Descriptive Analysis (QDA) technique contemplated in IRAM Standards 20,012 and 20,013 was used. These standards contemplate the analysis of the texture profile and procedures indicated in the "Guide D'Evaluation Sensorielle de la Texture des fromages a pâte dure ou semidure," 1994, of the European Union.

From the texture profile, the following descriptors were determined: (a) adherence, work that must be done with the tongue to detach a product stuck on the palate and the teeth; (b) solubility, a sensation that is highlighted when the sample melts very quickly in the saliva after chewing it four times with the molars; (c) humidity impression, perception of the moisture content of a food by means of tactile receptors in the mouth; (d) creaminess, a combination of properties such as viscosity, particle size, and lubrication; and (e) microstructure, related with geometric properties, particle size, shape, and arrangement, of the particles.

In the analysis of the flavor profile of cream cheeses, the following descriptors were evaluated, salty taste, sweet taste, bitter taste acid, and global persistence, relative to the response to a stimulus associated with a measurable period of time (IRAM 20001). Increasing continuous scales from 1 to 7 were used to express the intensity perceived in each descriptor. The panelists worked individually in booths analyzing in triplicate a number of two samples per session. Before performing the analysis, the samples were stabilized for 1 h at 9 ± 2°C and were presented inside sterilized glass vessels.

Data from the cheesemaking trials was statistically analyzed using the computer program "GraphPadInStat." The obtained data were statistically evaluated by the Tukey-Kramer multiple comparison test in the cases where two or more comparisons were considered. Otherwise, the t-test was used, assuming that a p < 0.05 was statistically significant [11].

#### **3. Characterization of the skimming of raw milk by microfiltration**

The raw material analysis indicated that it has a pH of 6.89 ± 0.02 and the following composition in w/100 g, 3.50 ± 0.08 of proteins, 2.00 ± 0.18 of fat, 50.00 ± 0.30 of saccharides, and 0.70 ± 0.05 of ash.

The results obtained showed that a fat content reduction of 76% (w/v) was achieved during the first stage, confirming that milk skimming by microfiltration was effective to obtain samples reduced in fat. The conventional operation to perform the separation of fat globules from milk is centrifugation. The decantation and spontaneous coalescence of the fat globules on the surface of the milk is slow; for this reason it is necessary to accelerate it by means of centrifugal separators. However, this technique presents difficulties during the separation of the fatty phase and requires the application of a high mechanical force. In the case of MF, fats are retained in the membrane. Thus, filtration technology is an effective way to achieve superior quality and safety, without compromising the fundamental sensory characteristics of the product. Also elimination of unwanted ingredients, such as microorganisms or sediments, which have a negative effect on quality, can be done, improving the texture of the final product and increasing its duration. On the other hand, it can shorten the stages of production and increase the yield, allowing a high degree of selectivity, and its energy costs are reduced. Similar results were reported in Refs. [12, 13].

#### **4. Physicochemical and sensory characterization of cream cheeses with and without FOS**

The results of the physicochemical characterization of the products are shown in **Table 1**. As expected, the highest change was in the fat cheese composition, between the full-fat cheese control and the rest of samples (p < 0.05). The codex general standard for cheese [14] determined that the FFCC obtained corresponds to a medium-fat cheese (25–45% w/w fat on dry basis) and the rest of the samples developed, to a low-fat cheese (10–25% w/w fat on dry basis). As the fat content

**91**

*Development of Functional Cheeses with Fructooligosaccharides*

**Proteins (%w/w)**

FFCC 46.05 ± 2.5 23.06 ± 1.5 24.07 ± 3.45 3.8 ± 0.32 3.02 ± 1.05 LFCC 48.95 ± 4.35 26.18 ± 1.01 15.83 ± 1.17 4.05 ± 0.35 4.99 ± 1.15

49.09 ± 1.35 25.52 ± 1.02 13.98 ± 0.40 3.89 ± 0.44 7.52 ± 0.50

49.01 ± 1.46 26.37 ± 2.17 14.16 ± 2.69 4.27 ± 0.47 6.144 ± 2.46

48.14 ± 1.67 28.45 ± 0.76 12.98 ± 0.55 4.05 ± 0.1 9 6.38 ± 1.92

47.46 ± 1.90 30.58 0.54 12.52 ± 0.14 3.67 ± 0.13 5.77 ± 0.72

**Fat (%w/w) Ash (%w/w) Carbohydrates** 

**(%w/w)**

of cheese is lowered, moisture content increases, and protein plays a greater role in texture development [15]. Thus, according to the Argentinean legislation [16], the samples correspond to cheeses of high moisture (46–54.9% w/w of moisture

*Physicochemical characterization of cream cheeses with and without FOS, at different concentrations (means ± SD).*

The samples M3 and M4, with inulin added as gel in different steps of cheese-

The samples with fructans showed an appropriate protein retention, higher than other results reported in Ref. [17], being the formulations with native fructans which retained the higher amount of fructans (M5 and M6), where a higher concentration of the NF affects greater protein retention (p < 0.05). This difference is attributed to the fact that, as mentioned, these fructans have a branched structure that can contribute better to forming a protein-saccharide network. The role of the agave fructans in the cheese matrix is significant, taking into account that they are considered as soluble fiber from natural and abundant sources, categorized as prebiotics. Thus, they become a valuable alternative as a functional ingredient in

Regarding the carbohydrate values, the amount of specific FOS retained by the

The results showed that the amount of inulin retained in M1 and M2 was very low probably due to its high solubility. The inulin incorporated as a gel at different times of the elaboration stage showed better results in terms of retention; however, it did not meet the expectations for the formulation of a functional type of cheese. Greater retention was achieved in the case of native agave fructan, probably as mentioned by the type of structure and lower solubility that allowed obtaining a microstructure with the proteins very similar to that of the whole control cheese

Control samples and cheeses with fructans showed high L\* value (> 80) which indicates the degree of luminosity; this parameter has a greater impact on the perceived appearance of the product. The values obtained were similar to other soft cheeses reported in the literature [18]. The values of a\* were positive close to zero without presenting significant differences between the samples (p > 0.05). The positive value of b\* indicated the degree of yellow hue. These values are adequate considering that they are low-fat cheeses and the elimination of fat imparts a translucent appearance. In effect, the colorimetric parameters obtained were in the order

content) and reduced in fat (10.0–24.9% w/w of fat content).

order to obtain functional foods.

sample, as was reported by SEM studies by [2].

making (**Figure 2**), have similar composition with that of M2 samples.

matrix was determined by HPLC. The results are presented in **Table 2**.

The color determination in the samples is presented in **Table 3**.

*DOI: http://dx.doi.org/10.5772/intechopen.85888*

**(%w/w)**

**Sample Moisture** 

M1-inuline (5%)

M2-inuline (1.5%)

M5-agave fructan (0.5%)

M6-agave fructan (5%)

**Table 1.**

*Development of Functional Cheeses with Fructooligosaccharides DOI: http://dx.doi.org/10.5772/intechopen.85888*


**Table 1.**

*Current Issues and Challenges in the Dairy Industry*

particles.

sterilized glass vessels.

statistically significant [11].

reported in Refs. [12, 13].

**with and without FOS**

molars; (c) humidity impression, perception of the moisture content of a food by means of tactile receptors in the mouth; (d) creaminess, a combination of properties such as viscosity, particle size, and lubrication; and (e) microstructure, related with geometric properties, particle size, shape, and arrangement, of the

In the analysis of the flavor profile of cream cheeses, the following descriptors were evaluated, salty taste, sweet taste, bitter taste acid, and global persistence, relative to the response to a stimulus associated with a measurable period of time (IRAM 20001). Increasing continuous scales from 1 to 7 were used to express the intensity perceived in each descriptor. The panelists worked individually in booths analyzing in triplicate a number of two samples per session. Before performing the analysis, the samples were stabilized for 1 h at 9 ± 2°C and were presented inside

Data from the cheesemaking trials was statistically analyzed using the computer program "GraphPadInStat." The obtained data were statistically evaluated by the Tukey-Kramer multiple comparison test in the cases where two or more comparisons were considered. Otherwise, the t-test was used, assuming that a p < 0.05 was

**3. Characterization of the skimming of raw milk by microfiltration**

50.00 ± 0.30 of saccharides, and 0.70 ± 0.05 of ash.

The raw material analysis indicated that it has a pH of 6.89 ± 0.02 and the following composition in w/100 g, 3.50 ± 0.08 of proteins, 2.00 ± 0.18 of fat,

The results obtained showed that a fat content reduction of 76% (w/v) was achieved during the first stage, confirming that milk skimming by microfiltration was effective to obtain samples reduced in fat. The conventional operation to perform the separation of fat globules from milk is centrifugation. The decantation and spontaneous coalescence of the fat globules on the surface of the milk is slow; for this reason it is necessary to accelerate it by means of centrifugal separators. However, this technique presents difficulties during the separation of the fatty phase and requires the application of a high mechanical force. In the case of MF, fats are retained in the membrane. Thus, filtration technology is an effective way to achieve superior quality and safety, without compromising the fundamental sensory characteristics of the product. Also elimination of unwanted ingredients, such as microorganisms or sediments, which have a negative effect on quality, can be done, improving the texture of the final product and increasing its duration. On the other hand, it can shorten the stages of production and increase the yield, allowing a high degree of selectivity, and its energy costs are reduced. Similar results were

**4. Physicochemical and sensory characterization of cream cheeses** 

The results of the physicochemical characterization of the products are shown in **Table 1**. As expected, the highest change was in the fat cheese composition, between the full-fat cheese control and the rest of samples (p < 0.05). The codex general standard for cheese [14] determined that the FFCC obtained corresponds to a medium-fat cheese (25–45% w/w fat on dry basis) and the rest of the samples developed, to a low-fat cheese (10–25% w/w fat on dry basis). As the fat content

**90**

*Physicochemical characterization of cream cheeses with and without FOS, at different concentrations (means ± SD).*

of cheese is lowered, moisture content increases, and protein plays a greater role in texture development [15]. Thus, according to the Argentinean legislation [16], the samples correspond to cheeses of high moisture (46–54.9% w/w of moisture content) and reduced in fat (10.0–24.9% w/w of fat content).

The samples M3 and M4, with inulin added as gel in different steps of cheesemaking (**Figure 2**), have similar composition with that of M2 samples.

The samples with fructans showed an appropriate protein retention, higher than other results reported in Ref. [17], being the formulations with native fructans which retained the higher amount of fructans (M5 and M6), where a higher concentration of the NF affects greater protein retention (p < 0.05). This difference is attributed to the fact that, as mentioned, these fructans have a branched structure that can contribute better to forming a protein-saccharide network. The role of the agave fructans in the cheese matrix is significant, taking into account that they are considered as soluble fiber from natural and abundant sources, categorized as prebiotics. Thus, they become a valuable alternative as a functional ingredient in order to obtain functional foods.

Regarding the carbohydrate values, the amount of specific FOS retained by the matrix was determined by HPLC. The results are presented in **Table 2**.

The results showed that the amount of inulin retained in M1 and M2 was very low probably due to its high solubility. The inulin incorporated as a gel at different times of the elaboration stage showed better results in terms of retention; however, it did not meet the expectations for the formulation of a functional type of cheese. Greater retention was achieved in the case of native agave fructan, probably as mentioned by the type of structure and lower solubility that allowed obtaining a microstructure with the proteins very similar to that of the whole control cheese sample, as was reported by SEM studies by [2].

The color determination in the samples is presented in **Table 3**.

Control samples and cheeses with fructans showed high L\* value (> 80) which indicates the degree of luminosity; this parameter has a greater impact on the perceived appearance of the product. The values obtained were similar to other soft cheeses reported in the literature [18]. The values of a\* were positive close to zero without presenting significant differences between the samples (p > 0.05). The positive value of b\* indicated the degree of yellow hue. These values are adequate considering that they are low-fat cheeses and the elimination of fat imparts a translucent appearance. In effect, the colorimetric parameters obtained were in the order

#### *Current Issues and Challenges in the Dairy Industry*


#### **Table 2.**

*Determination of FOS in the samples of cheese (means ± SD).*


#### **Table 3.**

*Surface cheese-like product color (means ± SD).*

of those reported for low-fat cheeses without coloring [19]. The results verified that the presence of fructans did not significantly affect the color of the samples with respect to the control cheese.

The sensorial analysis indicates a good acceptation of all the products according to the concentration range employed and did not present significant difference with regard to the control samples, indicating that the FOS did not affect these parameters. Even though it would be difficult to mimic entirely a full-fat cheese after fat has been removed, the presence of fructans in reduced fat formulations suggests an acceptable likeness in relation to structure and general characteristics of the full-fat control cheese. This fact constitutes a technological challenge.

#### **5. Characterization of soft cheeses with added inulin**

#### **5.1 Physicochemical composition**

**Table 4** shows the physicochemical composition of the soft cheese samples carried out by the process described in **Figure 4**, with inulin GR and HP added at 3% and 5% (w/v), which were also compared with a control samples without the polysaccharide (LFCC).

Each of these determinations was carried out in duplicate following the methodologies described for cream cheeses.

The results show that the cheeses are of high-moisture, higher than 70% w/w and low in fat minor of 11% w/w. In the case of carbohydrates, it is demonstrated that the addition of inulin was effectively retained in the cheese matrix, in

**93**

parameters.

in **Table 5**.

**Table 5.**

*\*Detection limit: 0.01 g/100 g*

**Table 4.**

serum corresponding to that sample.

*Inulin determination in soft cheese by HPLC (mean ± SD).*

of INTI-Dairy are presented in **Table 6**.

and that the cheeses were effectively enriched in fibers.

**5.2 Sensory evaluation of soft cheese with and without inulin**

*Development of Functional Cheeses with Fructooligosaccharides*

**Fat (%w/w) Protein** 

LFCC 74.06 ± 0.88 10.90 ± 0.14 10.79 ± 1.28 3.61 ± 0.57 0.65 ± 0.02 GR3 73.54 ± 3.31 10.23 ± 1.91 10.21 ± 1.75 6.52 ± 0.46 0.61 ± 0.03 GR5 73.56 ± 2.59 9.50 ± 1.41 8.57 ± 1.58 7.76 ± 0.42 0.63 ± 0.00 HP5 74.48 ± 1.20 10.80 ± 1.33 9.48 ± 0.99 5.36 ± 0.40 0.68 ± 0.00

*Physicochemical characterization of soft cheeses with and without FOS, at different concentrations (means ± SD).*

**Sample Inulin (%w/w)\*** C 0 ± 0.00 GR3 2.91 ± 0.13 GR5 2.84 ± 0.19 HP 0 ± 0.00 HP (in whey) 5.29 ± 0.01

**(%w/w)**

**Carbohydrates (%w/w)**

**Ash (%w/w)**

particular for the GR inulin since the increments found correspond to the quantities actually added. The determination by HPLC corroborated this result as it is shown

Cream cheese samples with inulin GR showed a retention of almost 100% of the added inulin. However, the HP inulin added to the sample was completely lost in the

The AFC (Chap. XVII, Art. 1386) indicates that for a food to be considered with added fiber, it must have at least 3 g/100 g in the case of solid foods and 1 g/100 ml of liquid foods. In the USA, the recommended daily consumption is 1–4 g/day, while in Europe a consumption of 3–11 g/day is suggested. Taking into account the results, we can say that the inulin content is high with respect to the recommended amounts

The results obtained in the sensory evaluation carried out by the trained panel

In this analysis, a sample of commercial spreadable cheese (CC), with similar characteristics, without fructans was added. The results of the sensory analysis indicated significant differences in the sweet flavor attribute and in the texture attribute creaminess and microstructure. The rest of the parameters analyzed did not show significant differences. Regarding the sweet taste, the difference found between the control sample (LFCC) and the sample with 5% inulin GR (GR5) may be due to the sweetening power of inulin, which, although much lower than that of sucrose, is considered a natural sweetener and when increasing the concentration it seems that it begins to be noticed. The differences found in the creaminess and the microstructure may be due to the kneading conditions, as it is known that this mechanical treatment significantly influences the expression of the aforementioned

*DOI: http://dx.doi.org/10.5772/intechopen.85888*

**(%w/w)**

**Sample Moisture** 



**Table 4.**

*Current Issues and Challenges in the Dairy Industry*

*Determination of FOS in the samples of cheese (means ± SD).*

respect to the control cheese.

*Surface cheese-like product color (means ± SD).*

**Table 3.**

**Table 2.**

**5.1 Physicochemical composition**

ologies described for cream cheeses.

polysaccharide (LFCC).

of those reported for low-fat cheeses without coloring [19]. The results verified that the presence of fructans did not significantly affect the color of the samples with

**Sample L a b** FFCC 85.30 ± 2.14 1.20 ± 0.11 18.37 ± 2.43 LFCC 86.11 ± 3.41 0.19 ± 0.16 14.89 ± 2.57 M1-inuline (5%) 84.58 ± 2.12 0.57 ± 1.12 17.32 ± 0.6 5 M2-inuline (1.5%) 82.26 ± 2.14 0.27 ± 0.15 16.90 ± 1.14 FN 0.5 81.50 ± 1.01 0.48 ± 0.18 16.32 ± 1.13 FN 5 89.60 ± 1.14 0.34 ± 0.09 16.66 ± 0.45

**Sample Inulin (%w/w)** FFCC — LFCC — M1-inuline (5%) 1.04 ± 0.11 M2-inuline (1.5%) 0.65 ± 0.09 M3-inuline gel 1.80 ± 0.11 M4-inuline gel 1.30 ± 0.17 M5-agave fructan (0.5%) 0.90 ± 0.11 M6-agave fructan (5%) 4.12 ± 0.15

The sensorial analysis indicates a good acceptation of all the products according to the concentration range employed and did not present significant difference with regard to the control samples, indicating that the FOS did not affect these parameters. Even though it would be difficult to mimic entirely a full-fat cheese after fat has been removed, the presence of fructans in reduced fat formulations suggests an acceptable likeness in relation to structure and general characteristics of the full-fat

**Table 4** shows the physicochemical composition of the soft cheese samples carried out by the process described in **Figure 4**, with inulin GR and HP added at 3% and 5% (w/v), which were also compared with a control samples without the

Each of these determinations was carried out in duplicate following the method-

The results show that the cheeses are of high-moisture, higher than 70% w/w and low in fat minor of 11% w/w. In the case of carbohydrates, it is demonstrated that the addition of inulin was effectively retained in the cheese matrix, in

control cheese. This fact constitutes a technological challenge.

**5. Characterization of soft cheeses with added inulin**

**92**

*Physicochemical characterization of soft cheeses with and without FOS, at different concentrations (means ± SD).*


#### **Table 5.**

*Inulin determination in soft cheese by HPLC (mean ± SD).*

particular for the GR inulin since the increments found correspond to the quantities actually added. The determination by HPLC corroborated this result as it is shown in **Table 5**.

Cream cheese samples with inulin GR showed a retention of almost 100% of the added inulin. However, the HP inulin added to the sample was completely lost in the serum corresponding to that sample.

The AFC (Chap. XVII, Art. 1386) indicates that for a food to be considered with added fiber, it must have at least 3 g/100 g in the case of solid foods and 1 g/100 ml of liquid foods. In the USA, the recommended daily consumption is 1–4 g/day, while in Europe a consumption of 3–11 g/day is suggested. Taking into account the results, we can say that the inulin content is high with respect to the recommended amounts and that the cheeses were effectively enriched in fibers.

#### **5.2 Sensory evaluation of soft cheese with and without inulin**

The results obtained in the sensory evaluation carried out by the trained panel of INTI-Dairy are presented in **Table 6**.

In this analysis, a sample of commercial spreadable cheese (CC), with similar characteristics, without fructans was added. The results of the sensory analysis indicated significant differences in the sweet flavor attribute and in the texture attribute creaminess and microstructure. The rest of the parameters analyzed did not show significant differences. Regarding the sweet taste, the difference found between the control sample (LFCC) and the sample with 5% inulin GR (GR5) may be due to the sweetening power of inulin, which, although much lower than that of sucrose, is considered a natural sweetener and when increasing the concentration it seems that it begins to be noticed. The differences found in the creaminess and the microstructure may be due to the kneading conditions, as it is known that this mechanical treatment significantly influences the expression of the aforementioned parameters.



### **Table 6.**

**95**

**6. Conclusions**

**Figure 6.**

**Table 7.**

*Development of Functional Cheeses with Fructooligosaccharides*

*Box plots for the sweet taste attribute of the spreadable soft cheeses.*

*Instrumental texture of soft cheeses with and without inulin (mean ± SD).*

Additionally, a statistical analysis of boxes and mustache or box plots was made for the different attributes. **Figure 6** shows the graph obtained for the sweet taste, where the significant difference between the control sample and the GR5 inulin

**Sample Adhesion (g/sec) Fmax (g) Elasticity Workmax (g/sec)** LFCC 88.28 ± 6.15 20.41 ± 1.09 9.96 ± 0.06 151.24 ± 10.31 GR3 86.21 ± 3.61 15.77 ± 0.88 9.96 ± 0.02 114.48 ± 4.70 HP5 83.78 ± ±8.74 16.57 ± 0.50 9.97 ± 0.03 118.95 ± 7.35

The data obtained for instrumental analysis texture is presented in **Table 7**. This

Significant differences were found in the maximum work values between the cheese samples with inulin and the control sample. In this sense, inulin in general modifies the hardness of foods, increasing this parameter according to the food formulation in which it is applied, as was previously shown [20]. However, in the hardness results of the samples analyzed, they show a decrease in this parameter. This may be due to the fact that the presence of serum proteins disturbs the fine structure that is formed between the inulin crystals by Van der Waals bonds, allow-

In this chapter, different experiments of incorporation of fructans in cheesemaking matrices were carried out, trying to obtain a reduced fat cheese additive with compounds that behave like prebiotics and act as dietary fiber. This search led to the testing the incorporation of inulins of different origin and degree of polymer-

ing the gel formation, generating a decrease in hardness [7].

ization to two types of cheese: soft and cream cheese.

*DOI: http://dx.doi.org/10.5772/intechopen.85888*

sample can be seen more clearly.

analysis was performed in triplicate.

*Sensory analysis results of soft cheeses with inulin GR and HP (mean ± SD).*

#### **Figure 6.** *Box plots for the sweet taste attribute of the spreadable soft cheeses.*


#### **Table 7.**

*Current Issues and Challenges in the Dairy Industry*

**94**

**Sample**

LFCC

CC GR3 GR5 HP5 **Table 6.**

*Sensory analysis results of soft cheeses with inulin GR and HP (mean ± SD).*

2.25a,b ± 0.86

2.5 ± 0.85

4.38 ± 1.24 *The supra-indices a, b, and c indicate significant differences between the results in the same column (p < 0.05).*

3.13 ± 0.91

3.63 ± 1.22

3.29 ± 1.78

4.94a ± 1.21

3.60 ± 1.31

3.63b ± 1.22

2.19 ± 0.61

4.00 ± 0.87

2.56 ± 0.68

3.25 ± 0.97

3.06 ± 2.01

5.00a ± 1.22

3.81 ± 1.27

2.50a,b ± 1.31

2.06 ± 0.95

4.00 ± 0.87

3.31 ± 0.66

3.69 ± 1.48

1.94 ± 1.13

3.06b ± 1.33

2.50a,b ± 0.93

2.63 ± 1.11

4.56 ± 1.11

3.44 ± 0.46

4.00 ± 1,41

3.75 ± 2.22

3.81a,b ± 0.86

2.06a ± 0.78

2.38 ± 0.99

4.19 ± 0.75

2.63 ± 0.86

3.75 ± 0.83

3.13 ± 1.83

5.13a ± 1.36

**Sweet**

**Salad**

**Acid**

**Bitter**

**Persistence**

**Adherence**

**Creaminess**

**Moisture Impression**

3.25 ± 1.48 3.56 ± 1.26 4.13 ± 1.62

**Microstructure**

2.06a ± 1.18 2.75a,c ± 1.09 5.25b ± 0.97 1.94a ± 0.81 3.69c ± 0.94

5.19 ± 1.22

5.38 ± 1.19

5.25 ± 1.58

5.50 ± 1.31

5.63 ± 0.92

**Solubility**

*Instrumental texture of soft cheeses with and without inulin (mean ± SD).*

Additionally, a statistical analysis of boxes and mustache or box plots was made for the different attributes. **Figure 6** shows the graph obtained for the sweet taste, where the significant difference between the control sample and the GR5 inulin sample can be seen more clearly.

The data obtained for instrumental analysis texture is presented in **Table 7**. This analysis was performed in triplicate.

Significant differences were found in the maximum work values between the cheese samples with inulin and the control sample. In this sense, inulin in general modifies the hardness of foods, increasing this parameter according to the food formulation in which it is applied, as was previously shown [20]. However, in the hardness results of the samples analyzed, they show a decrease in this parameter.

This may be due to the fact that the presence of serum proteins disturbs the fine structure that is formed between the inulin crystals by Van der Waals bonds, allowing the gel formation, generating a decrease in hardness [7].

#### **6. Conclusions**

In this chapter, different experiments of incorporation of fructans in cheesemaking matrices were carried out, trying to obtain a reduced fat cheese additive with compounds that behave like prebiotics and act as dietary fiber. This search led to the testing the incorporation of inulins of different origin and degree of polymerization to two types of cheese: soft and cream cheese.

From the wide experimentation carried out, it was possible to conclude that in the development of soft cheeses with inulin, in all cases the samples had a reduced fat content and high humidity. The texture and the micrographs showed adequate similarity with the control cheeses without the addition of inulin. However, the inulin retention was insufficient to have a food with the desired functional characteristics.

In the case of cheeses with agave fructans, a greater retention of the oligosaccharide was shown, given that it has a more branched structure that probably contributes to a better retention through the protein matrix. On the other hand, the determinations of color, texture, and sensory analysis did not show significant differences by the addition of the fructan. This conclusion is important since it opens the possibility of diversifying the uses of agave, a plant of rapid and widespread growth in America.

Finally, the experiments made with creamy cheeses, type spreads indicated that the composition of these samples responded to a high content of moisture and lowfat content, where proteins and carbohydrates adequately compensated the texture of the samples. But in this case, it was determined that GR inulin (native or shortchain inulin) was retained 100% in the cheese matrix, obtaining a product with the desired functional characteristics.

The sensory and texture profiles additionally showed that they are cheeses similar to the control samples with adequate parameters that make the product obtain a spreadable cheese with functional characteristics according to the needs of the market.

Even though it would be difficult to mimic entirely a full-fat cheese after fat has been weakened, the presence of FOS in reduced fat formulations suggests an acceptable likeness in relation to structure and general characteristics of the full-fat control cheese. This fact constitutes a technological challenge. The role of fructans in the cheese matrix is significant, taking into account that they are considered as soluble fiber from natural and abundant sources, categorized as prebiotics. Thus, they become a valuable alternative as a functional ingredient in order to obtain functional foods.

Future work should be carried out to confirm these findings and thus optimize the addition of fructans in different formulations of dairy products.

#### **Acknowledgements**

Financial support provided by the SCyT, UNSL (Project 2- 3114 and 2-0918), and PICT 2012-0155 (ANPCyT) and also fellowships of Diana Palatnik during 2012–2017, CONICET (Argentine), are gratefully acknowledged, as well as the staff of the National Institute of Industrial Technology, INTI-Dairy (Buenos Aires), for leading the internship of Eng. Diana Palatnik and providing the raw materials and analyses necessary for the development of spreadable cream cheese.

#### **Conflict of interest**

There is no conflict of interest in the work presented.

#### **Thanks**

The collaboration of Dr. Rosa I. Ortiz Basurto and Mg. Pamela Aldrete Herrera of Integral Research Laboratory in Food Technologic Institute of Tepic, Nayarit, Mexico, is recognized.

**97**

**Author details**

Diana Palatnik1

Haydée Montero2

and Mercedes E. Campderrós1

Buenos Aires, Argentina

, Noelia Rinaldoni1

, Germán Aranibar2

\*Address all correspondence to: mcampd@gmail.com

provided the original work is properly cited.

\*

, Diego Corrales2

1 Faculty of Chemistry, Biochemistry and Pharmacy, National University of San Luis, Chemical Technology Research Institute (INTEQUI), CONICET, San Luis, Argentina

2 Center of Research and Development in Food Criotechnology CIDCA (UNLP-CONICET—CIC), Faculty of Engineering, University of La Plata UNLP, La Plata,

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium,

, María L. Castells2

, María L. Rolon<sup>2</sup>

,

, Noemi Zaritzky2

*Development of Functional Cheeses with Fructooligosaccharides*

AOAC Association of Official Agricultural Chemists

HPLC high-performance liquid chromatography

IRAM Instituto Argentino de Normalización y Certificación

*DOI: http://dx.doi.org/10.5772/intechopen.85888*

AFC Argentine Food Code

FFCC full-fat control cheese FOS fructooligosaccharide GR native or short-chain inulin HP high-performance inulin

LFCC low-fat control cheese MF microfiltration NF native fructans

**Acronyms**

*Development of Functional Cheeses with Fructooligosaccharides DOI: http://dx.doi.org/10.5772/intechopen.85888*

### **Acronyms**

*Current Issues and Challenges in the Dairy Industry*

desired functional characteristics.

characteristics.

the market.

**Acknowledgements**

**Conflict of interest**

Mexico, is recognized.

From the wide experimentation carried out, it was possible to conclude that in the development of soft cheeses with inulin, in all cases the samples had a reduced fat content and high humidity. The texture and the micrographs showed adequate similarity with the control cheeses without the addition of inulin. However, the inulin retention was insufficient to have a food with the desired functional

In the case of cheeses with agave fructans, a greater retention of the oligosaccharide was shown, given that it has a more branched structure that probably contributes to a better retention through the protein matrix. On the other hand, the determinations of color, texture, and sensory analysis did not show significant differences by the addition of the fructan. This conclusion is important since it opens the possibility of diversifying the uses of agave, a plant of rapid and widespread growth in America. Finally, the experiments made with creamy cheeses, type spreads indicated that the composition of these samples responded to a high content of moisture and lowfat content, where proteins and carbohydrates adequately compensated the texture of the samples. But in this case, it was determined that GR inulin (native or shortchain inulin) was retained 100% in the cheese matrix, obtaining a product with the

The sensory and texture profiles additionally showed that they are cheeses similar to the control samples with adequate parameters that make the product obtain a spreadable cheese with functional characteristics according to the needs of

Even though it would be difficult to mimic entirely a full-fat cheese after fat has been weakened, the presence of FOS in reduced fat formulations suggests an acceptable likeness in relation to structure and general characteristics of the full-fat control cheese. This fact constitutes a technological challenge. The role of fructans in the cheese matrix is significant, taking into account that they are considered as soluble fiber from natural and abundant sources, categorized as prebiotics. Thus, they become a valuable alternative as a functional ingredient in order to obtain functional foods. Future work should be carried out to confirm these findings and thus optimize

Financial support provided by the SCyT, UNSL (Project 2- 3114 and 2-0918), and PICT 2012-0155 (ANPCyT) and also fellowships of Diana Palatnik during 2012–2017, CONICET (Argentine), are gratefully acknowledged, as well as the staff of the National Institute of Industrial Technology, INTI-Dairy (Buenos Aires), for leading the internship of Eng. Diana Palatnik and providing the raw materials and

The collaboration of Dr. Rosa I. Ortiz Basurto and Mg. Pamela Aldrete Herrera of Integral Research Laboratory in Food Technologic Institute of Tepic, Nayarit,

the addition of fructans in different formulations of dairy products.

analyses necessary for the development of spreadable cream cheese.

There is no conflict of interest in the work presented.

**96**

**Thanks**


### **Author details**

Diana Palatnik1 , Noelia Rinaldoni1 , Diego Corrales2 , María L. Rolon<sup>2</sup> , Haydée Montero2 , Germán Aranibar2 , María L. Castells2 , Noemi Zaritzky2 and Mercedes E. Campderrós1 \*

1 Faculty of Chemistry, Biochemistry and Pharmacy, National University of San Luis, Chemical Technology Research Institute (INTEQUI), CONICET, San Luis, Argentina

2 Center of Research and Development in Food Criotechnology CIDCA (UNLP-CONICET—CIC), Faculty of Engineering, University of La Plata UNLP, La Plata, Buenos Aires, Argentina

\*Address all correspondence to: mcampd@gmail.com

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

### **References**

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[2] Palatnik DR, Aldrete Herrera P, Rinaldoni AN, Ortiz Basurto RI, Campderrós ME. Development of reduced fat cheeses with the incorporation of agave fructans. International Journal of Dairy Technology. 2016;**69**:1-8. DOI: 10.1111/1471-0307.12334

[3] Sáyago-Ayerdi SG, Mateos R, Ortiz-Basurto RI, Largo C, Serrano J, Granado-Serrano AB, et al. Effects of consuming diets containing Agave tequilana dietary fibre and jamaica calyces on body weight gain and redox status in hypercholesterolemic rats. Food Chemistry. 2014;**148**:54-59. DOI: 10.1016/j.foodchem.2013.10.004

[4] Wang Y. Prebiotics: Present and future in food science and technology. Food Research International. 2009;**42**: 8-12. DOI: 10.1016/j.foodres.2008. 09.001

[5] Bosscher D, Van Loo J, Franck A. Inulin and oligofructose as functional ingredients to improve bone mineralization. International Dairy Journal. 2006;**16**:1092-1097. DOI: 10.1016/j.idairyj.2005.10.028

[6] Cassano A, Rastogi NK, Basile A. Membrane technologies for water treatment and reuse in the food and beverage industries. In: Basile A, Cassano A, Rastogi NK, editors. Advances in Membrane Tech. for Water Treatment. Cambridge, UK: Woodhead Publishing Series in Energy; 2015. pp. 551-580. DOI: 10.1016/ C2013-0-16469-0

[7] Glibowski P. Rheological properties and structure of inulin-whey protein

gels. International Dairy Journal. 2009;**19**:443-449. DOI: 10.1016/j. idairyj.2009.03.011

[8] Horwitz W, Latimer G, editors. Official Methods of Analysis of AOAC International—18th Edition. In: Revision 18th AOAC International. USA: Annual Book of ASTM Standards; 2010

[9] Zuleta A, Sambucetti ME. Inulin determination for food labeling. Journal of Agricultural and Food Chemistry. 2001;**49**:4570-4572. DOI: 10.1021/ jf010505o

[10] Meilgaard MC, Arbor A, Carr T, Civille GV. Sensory Evaluation Techniques. 4th ed. Boca Raton, Fla, London: CRC, Taylor & Francis; 2006. p. 488

[11] SAS. USER GUIDE: Statistic. Version. Cary, NC, USA: SAS Inst. Inc; 1989

[12] Rinaldoni AN, Campderrós ME, Menéndez C, Pérez Padilla A. Fractionation of skim-milk by an integrated membrane process for yogurt elaboration and lactose recuperation. International Journal of Food Engineering. 2009;**5**:1-17. DOI: 10.2202 /1556-3758.1531

[13] Mercier-Bouchard D, Benoit S, Doyen A, Britten M, Pouliot Y. Process efficiency of casein separation from milk using polymeric spiral-wound microfiltration membranes. Journal of Dairy Science. 2017;**100**:8838-8848. DOI: 10.3168/jds.2017-13015

[14] Kosikowski FV, Mistry VV. Cheese and fermented milk foods. In: Origins and Principles. Vol. 1. 3rd ed. Westport, USA: FV Kosikowski LLC; 1997. p. 728

**99**

*Development of Functional Cheeses with Fructooligosaccharides*

*DOI: http://dx.doi.org/10.5772/intechopen.85888*

[16] Argentian Food Code, Administración Nacional de Alimentos, Medicamentos y Tecnología Médica Capítulo VIII (Alimentos Lácteos). Available form: http://www.anmat.gov.ar/ alimentos/codigo/CAPITULO\_VIII.pdf

[17] Salvatore E, Pes M, Mazzarello V, Pirisi A. Replacement of fat with long-chain inulin in a fresh cheese made from caprine milk. International Dairy Journal. 2014;**34**:1-5. DOI: 10.1016/j.

[18] Juric M, Bertelsen G, Mortensen G, Petersen MA. Light induced colour and aroma changes in sliced, modified atmosphere packaged semi-hard cheeses. International Dairy Journal. 2003;**13**:239-249. PII: S 0958-6946

[19] Wadhwani R, McMahon DJ. Color of low-fat cheese influences flavor perception and consumer liking. Journal of Dairy Science. 2012;**95**:2336-2346.

[20] Rodríguez Furlán LT, Baracco Y, Lecot J, Zaritzky N, Campderrós M. Influence of hydrogenated oil as cocoa butter replacers in the development of sugar-free compound chocolates: Use of inulin as stabilizing agent. Food Chemistry. 2017;**217**:637-647. DOI: 10.1016/j.foodchem.2016.09.054

DOI: 10.3168/jds.2011-5142

idairyj.2013.07.007

(02)00156-5

[15] Mistry VV. Low fat cheese technology. International Dairy Journal. 2001;**11**:413-422. PII: S 0958-6946(01)00077-2

*Development of Functional Cheeses with Fructooligosaccharides DOI: http://dx.doi.org/10.5772/intechopen.85888*

[16] Argentian Food Code, Administración Nacional de Alimentos, Medicamentos y Tecnología Médica Capítulo VIII (Alimentos Lácteos). Available form: http://www.anmat.gov.ar/ alimentos/codigo/CAPITULO\_VIII.pdf

[17] Salvatore E, Pes M, Mazzarello V, Pirisi A. Replacement of fat with long-chain inulin in a fresh cheese made from caprine milk. International Dairy Journal. 2014;**34**:1-5. DOI: 10.1016/j. idairyj.2013.07.007

[18] Juric M, Bertelsen G, Mortensen G, Petersen MA. Light induced colour and aroma changes in sliced, modified atmosphere packaged semi-hard cheeses. International Dairy Journal. 2003;**13**:239-249. PII: S 0958-6946 (02)00156-5

[19] Wadhwani R, McMahon DJ. Color of low-fat cheese influences flavor perception and consumer liking. Journal of Dairy Science. 2012;**95**:2336-2346. DOI: 10.3168/jds.2011-5142

[20] Rodríguez Furlán LT, Baracco Y, Lecot J, Zaritzky N, Campderrós M. Influence of hydrogenated oil as cocoa butter replacers in the development of sugar-free compound chocolates: Use of inulin as stabilizing agent. Food Chemistry. 2017;**217**:637-647. DOI: 10.1016/j.foodchem.2016.09.054

**98**

09.001

*Current Issues and Challenges in the Dairy Industry*

gels. International Dairy Journal. 2009;**19**:443-449. DOI: 10.1016/j.

[8] Horwitz W, Latimer G, editors. Official Methods of Analysis of AOAC International—18th Edition. In:

Revision 18th AOAC International. USA: Annual Book of ASTM Standards; 2010

[9] Zuleta A, Sambucetti ME. Inulin determination for food labeling. Journal of Agricultural and Food Chemistry. 2001;**49**:4570-4572. DOI: 10.1021/

[10] Meilgaard MC, Arbor A, Carr T, Civille GV. Sensory Evaluation Techniques. 4th ed. Boca Raton, Fla, London: CRC, Taylor & Francis; 2006.

[11] SAS. USER GUIDE: Statistic. Version. Cary, NC, USA: SAS Inst. Inc;

[12] Rinaldoni AN, Campderrós ME, Menéndez C, Pérez Padilla A. Fractionation of skim-milk by an integrated membrane process for yogurt elaboration and lactose recuperation.

Engineering. 2009;**5**:1-17. DOI: 10.2202

[13] Mercier-Bouchard D, Benoit S, Doyen A, Britten M, Pouliot Y. Process efficiency of casein separation from milk using polymeric spiral-wound microfiltration membranes. Journal of Dairy Science. 2017;**100**:8838-8848.

International Journal of Food

DOI: 10.3168/jds.2017-13015

[15] Mistry VV. Low fat cheese technology. International Dairy Journal. 2001;**11**:413-422. PII: S

0958-6946(01)00077-2

[14] Kosikowski FV, Mistry VV. Cheese and fermented milk foods. In: Origins and Principles. Vol. 1. 3rd ed. Westport, USA: FV Kosikowski LLC; 1997. p. 728

/1556-3758.1531

idairyj.2009.03.011

jf010505o

p. 488

1989

[1] Juan B, Zamora A, Quintana F, Guamis B, Trujilla AJ. Effect of inulin addition on the sensorial properties of reduced-fat fresh cheese. International Journal of Dairy Technology. 2013;**66**:478-483. DOI:

[2] Palatnik DR, Aldrete Herrera P, Rinaldoni AN, Ortiz Basurto RI, Campderrós ME. Development of

[3] Sáyago-Ayerdi SG, Mateos R, Ortiz-Basurto RI, Largo C, Serrano J, Granado-Serrano AB, et al. Effects of consuming diets containing Agave tequilana dietary fibre and jamaica calyces on body weight gain and redox status in hypercholesterolemic rats. Food Chemistry. 2014;**148**:54-59. DOI: 10.1016/j.foodchem.2013.10.004

[4] Wang Y. Prebiotics: Present and future in food science and technology. Food Research International. 2009;**42**: 8-12. DOI: 10.1016/j.foodres.2008.

[5] Bosscher D, Van Loo J, Franck A. Inulin and oligofructose as functional

[6] Cassano A, Rastogi NK, Basile A. Membrane technologies for water treatment and reuse in the food and beverage industries. In: Basile A, Cassano A, Rastogi NK, editors. Advances in Membrane Tech. for Water Treatment. Cambridge, UK: Woodhead Publishing Series in Energy;

2015. pp. 551-580. DOI: 10.1016/

[7] Glibowski P. Rheological properties and structure of inulin-whey protein

C2013-0-16469-0

ingredients to improve bone mineralization. International Dairy Journal. 2006;**16**:1092-1097. DOI: 10.1016/j.idairyj.2005.10.028

reduced fat cheeses with the incorporation of agave fructans. International Journal of Dairy Technology. 2016;**69**:1-8. DOI:

10.1111/1471-0307.12057

10.1111/1471-0307.12334

**References**

**101**

≥108

**Chapter 7**

**Abstract**

**1. Introduction**

sufficient amounts [1, 2].

food and beverages.

sp. *bulgaricus*

Probiotic Characteristics and

Health Benefits of the Yogurt

*Albert Krastanov and Salam A. Ibrahim*

**Keywords:** yogurt, *L. bulgaricus*, probiotic, health benefits

Bacterium *Lactobacillus delbrueckii*

*Ayowole Oyeniran, Rabin Gyawali, Sulaiman O. Aljaloud,* 

Yogurt is a good source of several micronutrients and has played an important role in human nutrition. Consumption of yogurt has been shown to promote health benefits due to the presence of live bacteria. A number of human studies have demonstrated that yogurt contains viable bacteria, and especially *L. bulgaricus*, improve the health of the host and thus qualifies as a bona fide probiotic in its own right. In this chapter, we review the literature covering attributes of the yogurt bacterium *L. bulgaricus* that confirm its probiotic bacterial characteristics.

Yogurt, defined as the product of milk fermentation by *Lactobacillus delbrueckii* subsp. *bulgaricus* and *Streptococcus thermophilus*, has a long history of beneficial impact on the well-being of humans. As starter cultures for yogurt production, lactic acid bacteria (LAB) display symbiotic relations during their growth in milk medium. To meet the National Yogurt Association's criteria for "live and active culture yogurt," the final yogurt product must contain live LAB in amounts

 CFU/g at the time of manufacture, and the cultures must remain active up to the end of the stated shelf life. These live cultures are also considered to be probiotics because the cultures provide health benefits to the host when consumed in

Elie Metchnikoff can be credited as the progenitor of what has now become

improved and senility delayed by colonizing the gut with the host-friendly bacteria found in yogurt. In a market report published by Allied Market Research, the global probiotics market is expected to garner \$57.4 billion by 2022, registering a compound annual growth rate (CAGR) of 7.7% during the period 2016–2022. Asia-Pacific was the dominant probiotics market and is expected to be the leading contributor in global revenue due to its high level of adoption of probiotic based

a highly profitable industry, probiotics. He theorized that health could be

#### **Chapter 7**

## Probiotic Characteristics and Health Benefits of the Yogurt Bacterium *Lactobacillus delbrueckii* sp. *bulgaricus*

*Ayowole Oyeniran, Rabin Gyawali, Sulaiman O. Aljaloud, Albert Krastanov and Salam A. Ibrahim*

#### **Abstract**

Yogurt is a good source of several micronutrients and has played an important role in human nutrition. Consumption of yogurt has been shown to promote health benefits due to the presence of live bacteria. A number of human studies have demonstrated that yogurt contains viable bacteria, and especially *L. bulgaricus*, improve the health of the host and thus qualifies as a bona fide probiotic in its own right. In this chapter, we review the literature covering attributes of the yogurt bacterium *L. bulgaricus* that confirm its probiotic bacterial characteristics.

**Keywords:** yogurt, *L. bulgaricus*, probiotic, health benefits

#### **1. Introduction**

Yogurt, defined as the product of milk fermentation by *Lactobacillus delbrueckii* subsp. *bulgaricus* and *Streptococcus thermophilus*, has a long history of beneficial impact on the well-being of humans. As starter cultures for yogurt production, lactic acid bacteria (LAB) display symbiotic relations during their growth in milk medium. To meet the National Yogurt Association's criteria for "live and active culture yogurt," the final yogurt product must contain live LAB in amounts ≥108 CFU/g at the time of manufacture, and the cultures must remain active up to the end of the stated shelf life. These live cultures are also considered to be probiotics because the cultures provide health benefits to the host when consumed in sufficient amounts [1, 2].

Elie Metchnikoff can be credited as the progenitor of what has now become a highly profitable industry, probiotics. He theorized that health could be improved and senility delayed by colonizing the gut with the host-friendly bacteria found in yogurt. In a market report published by Allied Market Research, the global probiotics market is expected to garner \$57.4 billion by 2022, registering a compound annual growth rate (CAGR) of 7.7% during the period 2016–2022. Asia-Pacific was the dominant probiotics market and is expected to be the leading contributor in global revenue due to its high level of adoption of probiotic based food and beverages.

The definition of probiotics has evolved over the years due to some gray areas regarding the characteristics of a typical probiotic. The internationally endorsed definition of probiotics is "live microorganisms that, when administered in adequate amounts, confer a health benefit on the host" [3]. Modulation of the host's immune system and promotion of host defense are the most commonly supported benefits of probiotics consumption [3]. Most probiotics are lactic acid bacteria *Lactobacillus* sp., *Bifidobacterium* sp., and *Enterococcus* sp.; *Escherichia coli* strain Nissle 1917; the yeast *Saccharomyces boulardii*; some enterococci (*Enterococcus faecium* SF68); *Bacillus* sp., and *Clostridium butyricum* [4, 5, 54].

#### **2. Origin of** *Lactobacillus bulgaricus*

Elie Metchnikoff was a Russian biologist and Nobel Prize laureate who attributed the longevity of Bulgarians who were regular consumers of yogurt to the lactobacilli bacteria of yogurt. Metchnikoff's claims attracted wide attention to yogurt at that time. However, it was the Bulgarian graduate student Grigoroff [6] who first isolated and characterized this lactobacilli bacteria from the starter used in producing Kiselo Mlyako (Bulgarian Yogurt). Grigoroff named the bacterium '*Bacillus A'* or what is now recognized as *Lactobacillus bulgaricus* according to the Bergey's classification of bacteria. The origin and natural habitat of commercial *L. bulgaricus* strains may not have a definite answer despite its strong Bulgarian ties as countries like China, Mongolia, Russia, and Turkey also enjoy a long history of naturally fermented dairy products. A study by Song et al. [7] highlighted the uniqueness of *L bulgaricus* strains isolated from traditionally fermented milk products from some of the aforementioned countries.

Moreover, it appears *L. bulgaricus* is on a continuing evolutionary journey as it has adapted itself from a plant source to milk-rich environment [8]. Michaylova et al. [9] have been able to isolate and characterize *L. bulgaricus* from certain plant species (*Cornus mas*) obtained from four regions in Bulgaria. Yılmaz et al. [10] isolated *L. bulgaricus* from raw milk samples collected from different parts of Turkey, while *L. bulgaricus* was one of the isolates from raw milk samples obtained from four races of Algerian goats [11]. A study by Song et al. [7] illustrates the diversity of *L. bulgaricus* and the fact that it might not be an exclusive preserve of Bulgaria.

#### **2.1** *L. bulgaricus* **as a probiotic**

Elie Metchnikoff is regarded in some quarters as the grandfather of probiotics because of the profound observation he made at the beginning of the twentieth century, a time when the function of the gut flora was completely alien and unknown. Elie Metchnikoff realized that there was a link between regular consumption of lactic acid bacteria in fermented milk products to longevity and enhanced health in a certain group of Bulgarian people. He attributed this beneficial effect to the colonization and implantation of the Bulgarian bacillus which is now characterized as *L. bulgaricus.* Elie Metchnikoff believed that aging and diseases were caused by putrefaction of protein in the bowel by intestinal bacteria and that LAB were capable of inhibiting the growth of these putrefactive bacteria. He was so committed to the fact that fermented products could beneficially alter the microflora of the gut and prolong life that he drank sour milk fermented by lactic acid bacteria every day until his death [12–14].

*L. bulgaricus* has all the attributes of standard probiotic bacteria. It is crucial for probiotic strains to be able to colonize the intestine and survive passage through the upper gastrointestinal tract (GI) in order confer health benefits [15].

**103**

their toxins.

*Probiotic Characteristics and Health Benefits of the Yogurt Bacterium* Lactobacillus delbrueckii*…*

However, there are doubts about the adhesion and survival of *L. bulgaricus* after passage through the human gut. For example, *L. bulgaricus* are not native flora of mammals plus they do not have enough bile salt hydrolase genes and cannot synthesize mucin-binding proteins, all of which are important for survival in the GI [8]. However, regular consumption of yogurt fermented by *L. bulgaricus* may facilitate the colonization of these bacteria in the gut. Elli et al. [16] investigated the recovery of viable *L. delbrueckii* subsp. *bulgaricus* and *S. thermophilus* from the fecal samples of 20 healthy volunteers who were fed commercial yogurt for 1 week and found these bacteria present in the samples, suggesting that the bacteria, can survive transit in the gastrointestinal tract. Similarly, Mater et al. [17] had earlier established the survival of *L. bulgaricus* and *Streptococcus thermophilus* after passage through the gastrointestinal tract. A total of 37 out of 39 stool samples retrieved from 13 healthy subjects over a 12-day period yogurt intake contained viable *L. bulgaricus*. An encapsulated mixture of *L. bulgaricus* and *S. thermophilus* in chitosan and sodium alginate also survived in a simulated

In a study involving 61 elderly volunteers who were randomly assigned to receive either placebo or probiotics, Moro-Garcia et al. [19] evaluated the immunomodulatory capacity of *Lactobacillus delbrueckii* subsp. *bulgaricus* 848, strain isolated from a region of Bulgaria (Stara Planina) known for the longevity of its population [20]. A positive effect on the immune system was recorded in that study. Blood samples were taken at the beginning of the study and again after 3 and 6 months for the researcher to characterize the cell subpopulation, measured cytokines, quantified T cell receptor excision circles (TREC), and determined human β-defensin-2 (hBD-2) concentrations and human cytomegalovirus (CMV). The group that received *Lactobacillus delbrueckii* subsp. *bulgaricus* 848 had an increase in the percentage of NK cells, an improvement in the parameters defining the immune risk profile (IRP), and an increase in T cell subsets that are less differentiated. There was also a reduced concentration of pro-inflammatory cytokine interleukin-8 but an increase in the antimicrobial peptide hBD-2. In a similar study comparing the consumption of yogurt with milk in elderly subjects, yogurt fermented with *Lactobacillus delbrueckii* ssp. *bulgaricus* OLL1073R-1, a polysaccharide-producing lactic acid bacterial strain, was more effective in reducing the risk of catching the common cold. *Lactobacillus delbrueckii* ssp. *bulgaricus* OLL1073R-1 has been proven to have better effects on the immune system than other lactic acid bacteria. For example, the cell body and the immunostimulatory polysaccharides of these bacteria were identified responsible for the activation of biological defense mechanisms against pathogens such as viruses [21]. A recent study by Yamamoto et al. [22] corroborated the immunomodulatory effect of *Lactobacillus delbrueckii* ssp. *bulgaricus* OLL1073R-1. Thirty-seven elderly persons residing in a single nursing had their immunoglobulin A (IgA) levels increased after ingesting 112 g of the yogurt every morning for 12 weeks. IgA plays a critical role in the defense of mucous membranes against foreign antigens and pathogens, directly neutralizing the infectivity of pathogens and

Today's health-conscious consumers are increasingly aware of food content which is a driving force in the market for organic foods. For example, some consumers are motivated to take extra steps to avoid foods that contain chemical preservatives. Despite the fact that chemical preservatives are generally regarded as safe, the long-term side effects are unknown. Focus is thus being shifted to bio-preservation as an alternative. The use of LAB strains as a probiotic and bioprotective culture in fermented products has been widely studied. LAB have a major potential for use in biopreservation to extend shelf

*DOI: http://dx.doi.org/10.5772/intechopen.86939*

gastrointestinal tract [18].

*Probiotic Characteristics and Health Benefits of the Yogurt Bacterium* Lactobacillus delbrueckii*… DOI: http://dx.doi.org/10.5772/intechopen.86939*

However, there are doubts about the adhesion and survival of *L. bulgaricus* after passage through the human gut. For example, *L. bulgaricus* are not native flora of mammals plus they do not have enough bile salt hydrolase genes and cannot synthesize mucin-binding proteins, all of which are important for survival in the GI [8]. However, regular consumption of yogurt fermented by *L. bulgaricus* may facilitate the colonization of these bacteria in the gut. Elli et al. [16] investigated the recovery of viable *L. delbrueckii* subsp. *bulgaricus* and *S. thermophilus* from the fecal samples of 20 healthy volunteers who were fed commercial yogurt for 1 week and found these bacteria present in the samples, suggesting that the bacteria, can survive transit in the gastrointestinal tract. Similarly, Mater et al. [17] had earlier established the survival of *L. bulgaricus* and *Streptococcus thermophilus* after passage through the gastrointestinal tract. A total of 37 out of 39 stool samples retrieved from 13 healthy subjects over a 12-day period yogurt intake contained viable *L. bulgaricus*. An encapsulated mixture of *L. bulgaricus* and *S. thermophilus* in chitosan and sodium alginate also survived in a simulated gastrointestinal tract [18].

In a study involving 61 elderly volunteers who were randomly assigned to receive either placebo or probiotics, Moro-Garcia et al. [19] evaluated the immunomodulatory capacity of *Lactobacillus delbrueckii* subsp. *bulgaricus* 848, strain isolated from a region of Bulgaria (Stara Planina) known for the longevity of its population [20]. A positive effect on the immune system was recorded in that study. Blood samples were taken at the beginning of the study and again after 3 and 6 months for the researcher to characterize the cell subpopulation, measured cytokines, quantified T cell receptor excision circles (TREC), and determined human β-defensin-2 (hBD-2) concentrations and human cytomegalovirus (CMV). The group that received *Lactobacillus delbrueckii* subsp. *bulgaricus* 848 had an increase in the percentage of NK cells, an improvement in the parameters defining the immune risk profile (IRP), and an increase in T cell subsets that are less differentiated. There was also a reduced concentration of pro-inflammatory cytokine interleukin-8 but an increase in the antimicrobial peptide hBD-2. In a similar study comparing the consumption of yogurt with milk in elderly subjects, yogurt fermented with *Lactobacillus delbrueckii* ssp. *bulgaricus* OLL1073R-1, a polysaccharide-producing lactic acid bacterial strain, was more effective in reducing the risk of catching the common cold. *Lactobacillus delbrueckii* ssp. *bulgaricus* OLL1073R-1 has been proven to have better effects on the immune system than other lactic acid bacteria. For example, the cell body and the immunostimulatory polysaccharides of these bacteria were identified responsible for the activation of biological defense mechanisms against pathogens such as viruses [21]. A recent study by Yamamoto et al. [22] corroborated the immunomodulatory effect of *Lactobacillus delbrueckii* ssp. *bulgaricus* OLL1073R-1. Thirty-seven elderly persons residing in a single nursing had their immunoglobulin A (IgA) levels increased after ingesting 112 g of the yogurt every morning for 12 weeks. IgA plays a critical role in the defense of mucous membranes against foreign antigens and pathogens, directly neutralizing the infectivity of pathogens and their toxins.

Today's health-conscious consumers are increasingly aware of food content which is a driving force in the market for organic foods. For example, some consumers are motivated to take extra steps to avoid foods that contain chemical preservatives. Despite the fact that chemical preservatives are generally regarded as safe, the long-term side effects are unknown. Focus is thus being shifted to bio-preservation as an alternative. The use of LAB strains as a probiotic and bioprotective culture in fermented products has been widely studied. LAB have a major potential for use in biopreservation to extend shelf

*Current Issues and Challenges in the Dairy Industry*

**2. Origin of** *Lactobacillus bulgaricus*

of the aforementioned countries.

**2.1** *L. bulgaricus* **as a probiotic**

day until his death [12–14].

The definition of probiotics has evolved over the years due to some gray areas regarding the characteristics of a typical probiotic. The internationally endorsed definition of probiotics is "live microorganisms that, when administered in adequate amounts, confer a health benefit on the host" [3]. Modulation of the host's immune system and promotion of host defense are the most commonly supported benefits of probiotics consumption [3]. Most probiotics are lactic acid bacteria *Lactobacillus* sp., *Bifidobacterium* sp., and *Enterococcus* sp.; *Escherichia coli* strain Nissle 1917; the yeast *Saccharomyces boulardii*; some enterococci (*Enterococcus* 

Elie Metchnikoff was a Russian biologist and Nobel Prize laureate who attributed the longevity of Bulgarians who were regular consumers of yogurt to the lactobacilli bacteria of yogurt. Metchnikoff's claims attracted wide attention to yogurt at that time. However, it was the Bulgarian graduate student Grigoroff [6] who first isolated and characterized this lactobacilli bacteria from the starter used in producing Kiselo Mlyako (Bulgarian Yogurt). Grigoroff named the bacterium '*Bacillus A'* or what is now recognized as *Lactobacillus bulgaricus* according to the Bergey's classification of bacteria. The origin and natural habitat of commercial *L. bulgaricus* strains may not have a definite answer despite its strong Bulgarian ties as countries like China, Mongolia, Russia, and Turkey also enjoy a long history of naturally fermented dairy products. A study by Song et al. [7] highlighted the uniqueness of *L bulgaricus* strains isolated from traditionally fermented milk products from some

Moreover, it appears *L. bulgaricus* is on a continuing evolutionary journey as it has adapted itself from a plant source to milk-rich environment [8]. Michaylova et al. [9] have been able to isolate and characterize *L. bulgaricus* from certain plant species (*Cornus mas*) obtained from four regions in Bulgaria. Yılmaz et al. [10] isolated *L. bulgaricus* from raw milk samples collected from different parts of Turkey, while *L. bulgaricus* was one of the isolates from raw milk samples obtained from four races of Algerian goats [11]. A study by Song et al. [7] illustrates the diversity of *L. bulgaricus* and the fact that it might not be an exclusive preserve of Bulgaria.

Elie Metchnikoff is regarded in some quarters as the grandfather of probiotics because of the profound observation he made at the beginning of the twentieth century, a time when the function of the gut flora was completely alien and unknown. Elie Metchnikoff realized that there was a link between regular consumption of lactic acid bacteria in fermented milk products to longevity and enhanced health in a certain group of Bulgarian people. He attributed this beneficial effect to the colonization and implantation of the Bulgarian bacillus which is now characterized as *L. bulgaricus.* Elie Metchnikoff believed that aging and diseases were caused by putrefaction of protein in the bowel by intestinal bacteria and that LAB were capable of inhibiting the growth of these putrefactive bacteria. He was so committed to the fact that fermented products could beneficially alter the microflora of the gut and prolong life that he drank sour milk fermented by lactic acid bacteria every

*L. bulgaricus* has all the attributes of standard probiotic bacteria. It is crucial

for probiotic strains to be able to colonize the intestine and survive passage through the upper gastrointestinal tract (GI) in order confer health benefits [15].

*faecium* SF68); *Bacillus* sp., and *Clostridium butyricum* [4, 5, 54].

**102**

life and enhance the safety of foods [23]. For example, metabolites and antimicrobial products obtained from LAB have inhibitory effect against spoilage microorganisms. Most importantly, LAB produce organic acid such as lactic acid that reduced the pH of the food, thereby inhibiting the growth of other microflora [24, 25]. In the case of yogurt, drop in pH alters the yogurt environment resulting in an unfavorable medium for the development of some pathogens and spoilage microorganisms [26]. For example, strains of *Lactobacillus delbrueckii* ssp*. bulgaricus* and *Streptococcus thermophilus* isolated from Turkish homemade yogurt had inhibitory effects on *Escherichia coli* and *Listeria monocytogenes* [27].

The use of antimicrobial peptides such as bacteriocins that are produced by LAB will help to significantly minimize the use of chemical preservatives and could thus be used in hurdle technology to produce a more naturally preserved food. LAB produce bacteriocins, bioactive peptides or proteins, and bacteriocinlike inhibitory substances that are antimicrobial compounds that possess bacteriocin capacities requisites but have not been characterized for their amino acid sequence [24, 25]. Some *L. bulgaricus* strains which were isolated from yogurts had an antibacterial effect on *Vibrio cholerae* and *E. coli* due to significant bacteriocin production [28]. A study by Boyanova et al. [29] suggested that the bacteriocin-like inhibitory effects of GLB strains of *L. bulgaricus* could be valuable in the control of *Helicobacter pylori* infections. Clinical benefits were also reported in Thailand, where the addition of *L. delbrueckii* subsp. *bulgaricus* and *Streptococcus thermophilus* either before or before and after a 1-week tailored triple therapy regimen significantly improved eradication rates in *H. pylori infection* treatment [30].

Another area of diet concern in which probiotics come into play is the issue of lead (Pb) poisoning. Food and supplements or methods of food preparation may be a source of lead exposure that can have devastating effects on human health, and it remains a public health concern. *L. bulgaricus* KLDS1.0207 that has been isolated from traditional dairy products in Sinkiang Province, China has been evaluated for its protective effects against acute lead toxicity in mice. The high Pb-binding ability and high resistance to Pb in *L. bulgaricus* KLDS1.0207 offered protective effects from acute Pb toxicity in mice. The results in vivo demonstrated that this particular strain of *L. bulgaricus* can relieve renal pathological damage, reduce mortality rates, and enhance the antioxidant index in the liver and kidney making it a potential probiotic against lead toxicity [31, 32].

Another claimed health benefit linked to probiotics is an improvement in lactose metabolism ([33, 34]). It is widely agreed that fermented milk products such as yogurt can help with lactose digestion in lactose malabsorbers and therefore can be well tolerated by most lactose-intolerant subjects. Yogurt preparation using the traditional *S. thermophilus* and *L. delbrueckii* ssp. *bulgaricus* are even more effective due to their higher β-galactosidase activity. Lactose intolerance is a β-galactosidase deficiency resulting in the inability to digest lactose into the monosaccharides glucose and galactose. People with lactose intolerance develop diarrhea, abdominal discomfort, and flatulence after consumption of milk or milk products. Numerous studies have shown better lactose digestion and consequently less hydrogen exhalation in lactose malabsorbers who consumed yogurt with live cultures rather than with milk or pasteurized yogurt [35, 36].

All of these documented benefits and characteristics of probiotics, in general, provide an equally compelling argument for the effectiveness of *L. bulgaricus* as a probiotic. Yogurt remains one of the most important vehicles for the delivery of probiotic bacteria.

**105**

**Table 1.**

*Probiotic strains of* Lactobacillus bulgaricus.

*Probiotic Characteristics and Health Benefits of the Yogurt Bacterium* Lactobacillus delbrueckii*…*

**Table 1** lists *L. bulgaricus* strains that are beneficial for a range of health-related

*L. bulgaricus* **strain Probiotic activity References**

848 Immunomodulatory function [20]

influenza virus activity

KLDS1-0207 Protects against lead toxicity [31, 32]

bactericidal against *Helicobacter pylori*

Inhibitory action against periodontal pathogen; *Aggregatibacter actinomycetemcomitans*

*Porphyromonas gingivalis*, *A. actinomycetemcomitans*,

Inhibitory effects on *Salmonella* spp., *Pseudomonas* 

*Staphylococcus aureus*, *Pseudomonas fragi*, *Micrococcus* 

[37]

[38]

[40]

[41]

[42]

[43]

[28]

[45]

[46]

[48]

[54]

[27]

[21, 22, 39]

RTF Antibacterial activity against *Escherichia coli*,

*flavus*

7994 Inhibitory effect on *Achromobacter liquefaciens*, *S. aureus*, *P. fragi*

OLL1073R-1 Reduces risk of catching common cold, anti-

BB18 Production of bacteriocin (bulgaricin BB18);

B-30892 Inhibits *Clostridium difficile*-mediated cytotoxicity on Caco2 cells

Commercial yogurt isolate Inhibitory action against periodontal pathogens;

Commercial yogurt isolate Bacteriocin production inhibitory against *Vibrio cholerae* and *E. coli*

TLB06FT Antibacterial activity against *E. coli*, *S. aureus*,

CRL 871 Production of folate; an alternative to folic acid fortification

GB N1 (48) Hypolipidemic and protective cardiovascular effect

F5R Inhibitory effects on *Bacillus coagulans*, *B. cereus*,

*aureus* and *E. coli*

and *Prevotella nigrescens*

Commercial yogurt isolate Inhibitory effect on *E. coli* O157:H7 [44]

*aeruginosa*, *E. coli*, *S. aureus*

CRL 454 Aids digestion of allergenic β-lactoglobulin [47]

761 N Free radical scavenging ability; antiviral ability [49] GLB Antimicrobial; control of *H. pylori* [29]

D6R; PTCC 1332 Inhibitory effects on *S. aureus* and *E. coli* [27, 50]

DSM 20081 Inhibitory effect on *E*. *coli* [51, 52] DWT1 Inhibits tumor growth [53]

*P. fluorescens*, *K. pneumoniae*, *L. monocytogenes*, *S.* 

*P. aeruginosa*, *Listeria monocytogenes*

*DOI: http://dx.doi.org/10.5772/intechopen.86939*

ATCC 11 842, LBL-23, LBL-12, LBL-22, LBL-6, LBL-10, LBL-13, LBL-83, LBL-42, LBL-9,

NCTC 12197 Tat, DSMZ

20080 T

LBL-11

issues.

*Probiotic Characteristics and Health Benefits of the Yogurt Bacterium* Lactobacillus delbrueckii*… DOI: http://dx.doi.org/10.5772/intechopen.86939*

**Table 1** lists *L. bulgaricus* strains that are beneficial for a range of health-related issues.


#### **Table 1.**

*Current Issues and Challenges in the Dairy Industry*

*cytogenes* [27].

treatment [30].

probiotic against lead toxicity [31, 32].

with milk or pasteurized yogurt [35, 36].

life and enhance the safety of foods [23]. For example, metabolites and antimicrobial products obtained from LAB have inhibitory effect against spoilage microorganisms. Most importantly, LAB produce organic acid such as lactic acid that reduced the pH of the food, thereby inhibiting the growth of other microflora [24, 25]. In the case of yogurt, drop in pH alters the yogurt environment resulting in an unfavorable medium for the development of some pathogens and spoilage microorganisms [26]. For example, strains of *Lactobacillus delbrueckii* ssp*. bulgaricus* and *Streptococcus thermophilus* isolated from Turkish homemade yogurt had inhibitory effects on *Escherichia coli* and *Listeria mono-*

The use of antimicrobial peptides such as bacteriocins that are produced by LAB will help to significantly minimize the use of chemical preservatives and could thus be used in hurdle technology to produce a more naturally preserved food. LAB produce bacteriocins, bioactive peptides or proteins, and bacteriocinlike inhibitory substances that are antimicrobial compounds that possess bacteriocin capacities requisites but have not been characterized for their amino acid sequence [24, 25]. Some *L. bulgaricus* strains which were isolated from yogurts had an antibacterial effect on *Vibrio cholerae* and *E. coli* due to significant bacteriocin production [28]. A study by Boyanova et al. [29] suggested that the bacteriocin-like inhibitory effects of GLB strains of *L. bulgaricus* could be valuable in the control of *Helicobacter pylori* infections. Clinical benefits were also reported in Thailand, where the addition of *L. delbrueckii* subsp. *bulgaricus* and *Streptococcus thermophilus* either before or before and after a 1-week tailored triple therapy regimen significantly improved eradication rates in *H. pylori infection*

Another area of diet concern in which probiotics come into play is the issue of lead (Pb) poisoning. Food and supplements or methods of food preparation may be a source of lead exposure that can have devastating effects on human health, and it remains a public health concern. *L. bulgaricus* KLDS1.0207 that has been isolated from traditional dairy products in Sinkiang Province, China has been evaluated for its protective effects against acute lead toxicity in mice. The high Pb-binding ability and high resistance to Pb in *L. bulgaricus* KLDS1.0207 offered protective effects from acute Pb toxicity in mice. The results in vivo demonstrated that this particular strain of *L. bulgaricus* can relieve renal pathological damage, reduce mortality rates, and enhance the antioxidant index in the liver and kidney making it a potential

Another claimed health benefit linked to probiotics is an improvement in lactose

All of these documented benefits and characteristics of probiotics, in general, provide an equally compelling argument for the effectiveness of *L. bulgaricus* as a probiotic. Yogurt remains one of the most important vehicles for the delivery of

metabolism ([33, 34]). It is widely agreed that fermented milk products such as yogurt can help with lactose digestion in lactose malabsorbers and therefore can be well tolerated by most lactose-intolerant subjects. Yogurt preparation using the traditional *S. thermophilus* and *L. delbrueckii* ssp. *bulgaricus* are even more effective due to their higher β-galactosidase activity. Lactose intolerance is a β-galactosidase deficiency resulting in the inability to digest lactose into the monosaccharides glucose and galactose. People with lactose intolerance develop diarrhea, abdominal discomfort, and flatulence after consumption of milk or milk products. Numerous studies have shown better lactose digestion and consequently less hydrogen exhalation in lactose malabsorbers who consumed yogurt with live cultures rather than

**104**

probiotic bacteria.

*Probiotic strains of* Lactobacillus bulgaricus.

### **3. Conclusion**

Because of the documented health benefits conferred by *L. bulgaricus*, consumer demand for yogurt and yogurt-related products has recently become the fastest growing dairy category in the global market. Our literature review showed that *L. bulgaricus* clearly qualifies as a probiotic in its own right. This growing popularity is not surprising in light of the fact that Nobel Laureate Metchnikoff linked the health and longevity of the Bulgarian people to their high consumption of yogurt containing *L. bulgaricus*. Consequently, increased yogurt intake should be a promising addition to a healthy dietary regimen that led to health promotion and well-being. Future work should be directed to understand the metabolites produced by *L. bulgaricus* and their health benefits. With a more comprehensive understanding on the functional properties of *L. bulgaricus*, we could advocate the importance of yogurt consumption and its impact on our well-being.

### **Acknowledgements**

This publication was made possible by grant number NC.X-291-5-15-170-1 from the National Institute of Food and Agriculture (NIFA) through the Agricultural Research program at North Carolina A&T State University and in part by the Indiana University and Jarrow Formulas, USA.

### **Author details**

Ayowole Oyeniran1 , Rabin Gyawali1 , Sulaiman O. Aljaloud2 , Albert Krastanov3 and Salam A. Ibrahim1 \*

1 Food Microbiology and Biotechnology Laboratory, Food and Nutritional Sciences Program, North Carolina A&T State University, Greensboro, USA

2 Department of Exercise Physiology, College of Sport Sciences and Physical Activity, King Saud University, Riyadh, Saudi Arabia

3 Department of Biotechnology, University of Food Technologies, Plovdiv, Bulgaria

\*Address all correspondence to: ibrah001@ncat.edu

© 2020 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

**107**

*Probiotic Characteristics and Health Benefits of the Yogurt Bacterium* Lactobacillus delbrueckii*…*

[8] Van de Guchte M, Penaud S, Grimaldi C, Barbe V, Bryson K, Nicolas P, et al. The complete genome sequence of *Lactobacillus bulgaricus* reveals extensive and ongoing reductive evolution. Proceedings of the National Academy of Sciences of the United States of America. 2006;**103**(24):

9274-9279. DOI: 10.1073/

[9] Michaylova M, Minkova S, Kimura K, Sasaki T, Isawa K. Isolation and characterization of *Lactobacillus delbrueckii* ssp. *bulgaricus* and

*Streptococcus thermophilus* from plants in Bulgaria. FEMS Microbiology Letters. 2007;**269**(1):160-169. DOI: 10.1111/j.1574-6968.2007.00631.x

[10] Yılmaz R, Temiz A, Açık L, Çelebi Keskin A. Genetic differentiation of *Lactobacillus delbrueckii* subsp. *bulgaricus* and *Streptococcus thermophilus* strains isolated from raw milk samples collected from different regions of Turkey. Food Biotechnology. 2015;**29**(4):336-355. DOI: 10.1080/08905436.2015.1092091

[11] Badis A, Guetarni D, Moussa Boudjema B, Henni DE, Kihal M. Identification and technological properties of lactic acid bacteria isolated from raw goat milk of four Algerian races. Food Microbiology. 2004;**21**(5):579-588. DOI: 10.1016/j.

[12] Anukam KC, Reid G. Probiotics:

100 years (1907-2007) after Elie Metchnikoff's observation. Communicating Current Research and Educational Topics and Trends in Applied Microbiology. 2007;**1**:466-474

[13] Hawrelak JA, Myers SP. The causes of intestinal dysbiosis: A review. Alternative Medicine Review. 2004;**9**(2):180-197. DOI: 10.1016/0965-2299(93)90012-3

fm.2003.11.006

pnas.0603024103

*DOI: http://dx.doi.org/10.5772/intechopen.86939*

[1] Bourlioux P, Pochart P. Nutritional and health properties of yogurt. World Review of Nutrition and

[2] Chandan RC, Gandhi A, Shah NP. Yogurt: Historical background, health benefits, and global trade. In: Yogurt in Health and Disease Prevention. London: Academic Press; 2017. pp. 3-29. DOI: 10.1016/B978-0-12-805134-4.00001-8

[3] Gyawali R, Nwamaioha N, Fiagbor R, Zimmerman T, Newman RH, Ibrahim SA. The role of prebiotics in disease prevention and health promotion. In: Dietary Interventions in Gastrointestinal Diseases. United Kingdom: Academic Press; 2019.

[4] Elmer GW, Martin SW, Horner KL, Mcfarland LV, Levy RH. Survival of *Saccharomyces boulardii* in the rat gastrointestinal tract and effects of dietary fiber. Microbial Ecology in Health and Disease. 1999;**11**(1):29-34

[5] Takahashi M, Taguchi H, Yamaguchi H, Osaki T, Komatsu A, Kamiya S. The effect of probiotic treatment with *Clostridium butyricum* on enterohemorrhagic *Escherichia coli* O157: H7 infection in mice. FEMS Immunology and Medical Microbiology.

**References**

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pp. 151-167

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L'Université. 1905

10.1038/srep22704

[6] Grigoroff S. Étude sur une lait fermenté comestible. Le "Kissélo mléko" de Bulgarie. Revue Médicale de la Suisse Romande. Genéve: Georg&G;

LibrairesÉditeurs. Librairie de

[7] Song Y, Sun Z, Guo C, Wu Y, Liu W, Yu J, et al. Genetic diversity and population structure of *Lactobacillus delbrueckii* subspecies *bulgaricus* isolated from naturally fermented dairy foods. Scientific Reports. 2016;**6**:22704. DOI:

*Probiotic Characteristics and Health Benefits of the Yogurt Bacterium* Lactobacillus delbrueckii*… DOI: http://dx.doi.org/10.5772/intechopen.86939*

#### **References**

*Current Issues and Challenges in the Dairy Industry*

Because of the documented health benefits conferred by *L. bulgaricus*, consumer

This publication was made possible by grant number NC.X-291-5-15-170-1 from the National Institute of Food and Agriculture (NIFA) through the Agricultural Research program at North Carolina A&T State University and in part by the

, Sulaiman O. Aljaloud2

1 Food Microbiology and Biotechnology Laboratory, Food and Nutritional Sciences

3 Department of Biotechnology, University of Food Technologies, Plovdiv, Bulgaria

© 2020 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium,

2 Department of Exercise Physiology, College of Sport Sciences and Physical

, Albert Krastanov3

demand for yogurt and yogurt-related products has recently become the fastest growing dairy category in the global market. Our literature review showed that *L. bulgaricus* clearly qualifies as a probiotic in its own right. This growing popularity is not surprising in light of the fact that Nobel Laureate Metchnikoff linked the health and longevity of the Bulgarian people to their high consumption of yogurt containing *L. bulgaricus*. Consequently, increased yogurt intake should be a promising addition to a healthy dietary regimen that led to health promotion and well-being. Future work should be directed to understand the metabolites produced by *L. bulgaricus* and their health benefits. With a more comprehensive understanding on the functional properties of *L. bulgaricus*, we could advocate the importance of yogurt consumption and its

**3. Conclusion**

impact on our well-being.

**Acknowledgements**

**Author details**

Ayowole Oyeniran1

and Salam A. Ibrahim1

Indiana University and Jarrow Formulas, USA.

, Rabin Gyawali1

Activity, King Saud University, Riyadh, Saudi Arabia

\*Address all correspondence to: ibrah001@ncat.edu

provided the original work is properly cited.

Program, North Carolina A&T State University, Greensboro, USA

\*

**106**

[1] Bourlioux P, Pochart P. Nutritional and health properties of yogurt. World Review of Nutrition and Dietetics. 1988

[2] Chandan RC, Gandhi A, Shah NP. Yogurt: Historical background, health benefits, and global trade. In: Yogurt in Health and Disease Prevention. London: Academic Press; 2017. pp. 3-29. DOI: 10.1016/B978-0-12-805134-4.00001-8

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[17] Mater DDG, Bretigny L, Firmesse O, Flores M-J, Mogenet A, Bresson J-L, et al. *Streptococcus thermophilus* and *Lactobacillus delbrueckii* subsp. *bulgaricus* survive gastrointestinal transit of healthy volunteers consuming yogurt. FEMS Microbiology Letters. 2005;**250**(2):185-187. DOI: 10.1016/j. femsle.2005.07.006

[18] Vodnar DC, Socaciu C, Rotar AM, Stãnilã A. Morphology, FTIR fingerprint and survivability of encapsulated lactic bacteria (*Streptococcus thermophilus* and *Lactobacillus delbrueckii* subsp. *bulgaricus*) in simulated gastric juice and intestinal juice. International Journal of Food Science and Technology. 2010;**45**(11):2345-2351. DOI: 10.1111/j.1365-2621.2010.02406.x

[19] Moro-Garcia MA, Alonso-Arias R, Baltadjieva M, Benitez CF, Barrial MAF, Ruisánchez ED, et al. Oral supplementation with *Lactobacillus delbrueckii* subsp. *bulgaricus* 8481 enhances systemic immunity in elderly subjects. Age. 2013;**35**(4):1311-1326. DOI: 10.1007/s11357-012-9434-6

[20] Dixon B. Secrets of the Bulgarian bacillus. The Lancet Infectious Diseases. 2002;**2**(4):260

[21] Makino S, Ikegami S, Kume A, Horiuchi H, Sasaki H, Orii N. Reducing the risk of infection in the elderly by dietary intake of yoghurt fermented with *Lactobacillus delbrueckii* ssp. *bulgaricus* OLL1073R-1. British Journal of Nutrition. 2010;**104**(7):998-1006. DOI: 10.1017/S000711451000173X

[22] Yamamoto Y, Fujino K, Saruta J, Takahashi T, To M, Fuchida S, et al. Effects of yogurt fermented with *Lactobacillus delbrueckii* ssp. *bulgaricus* OLL1073R-1 on the IgA flow rate of saliva in elderly persons residing in a nursing home: A before-after non-randomised intervention study. Gerodontology. 2017;**34**(4):479- 485. DOI: 10.1111/ger.12296

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[25] Reis JA, Paula AT, Casarotti S, Penna A. Lactic acid bacteria antimicrobial compounds: Characteristics and applications. Food Engineering Reviews. 2012;**4**. DOI: 10.1007/s12393-012-9051-2

[26] Bachrouri M, Quinto EJ, Mora MT. Kinetic parameters of *Escherichia coli* O157: H7 survival during fermentation of milk and refrigeration of home-made yoghurt. International Dairy Journal. 2006;**16**(5):474-481

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[37] Singh J, Khanna A, Chander B. Antibacterial activity of yogurt starter in cow and buffalo milk. Journal of Food

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[39] Nagai T, Makino S, Ikegami S, Itoh H, Yamada H. Effects of oral administration of yogurt fermented

with *Lactobacillus delbrueckii* ssp. *bulgaricus* OLL1073R-1 and its exopolysaccharides against influenza virus infection in mice.

10.5402/2013/481651

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ajcn/73.2.421s

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*DOI: http://dx.doi.org/10.5772/intechopen.86939*

[28] Tufail M, Hussain S, Malik F, Mirza T, Parveen G, Shafaat S, et al. Isolation and evaluation of antibacterial activity of bacteriocin produced by *Lactobacillus* 

[29] Boyanova L, Gergova G, Markovska R, Yordanov D, Mitov I. Bacteriocinlike inhibitory activities of seven *Lactobacillus delbrueckii* subsp. *bulgaricus* strains against antibiotic susceptible and resistant *Helicobacter pylori* strains. Letters in Applied

Microbiology. 2017;**65**(6):469-474. DOI:

[30] Tongtawee T, Dechsukhum C, Leeanansaksiri W, Kaewpitoon S, Kaewpitoon N, Loyd RA, et al. Improved helicobacter pylori eradication rate of tailored triple therapy by adding *Lactobacillus delbrueckii* and *Streptococcus thermophilus* in northeast region of Thailand: A prospective

randomized controlled clinical trial. Gastroenterology Research and Practice. 2015;**2015**:518018. DOI:

[31] Fewtrell LJ, Prüss-Üstün A, Landrigan P, Ayuso-Mateos JL. Estimating the global burden of disease of mild mental retardation and cardiovascular diseases from environmental lead exposure. Environmental Research.

2004;**94**(2):120-133. DOI: 10.1016/

[32] Li B, Jin D, Yu S, Etareri Evivie S, Muhammad Z, Huo G, et al. In vitro and in vivo evaluation of *Lactobacillus* 

S0013-9351(03)00132-4

*bulgaricus* from yogurt. African Journal of Microbiology Research.

2011;**5**(22):3842-3847

10.1111/lam.12807

10.1155/2015/518018

resistance of *Lactobacillus delbrueckii* ssp. bulgaricus and *Streptococcus thermophilus* strains isolated from Turkish homemade yoghurts. African Journal of Microbiology Research. 2011;**5**(6):675-682. DOI: 10.5897/

AJMR10.835

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resistance of *Lactobacillus delbrueckii* ssp. bulgaricus and *Streptococcus thermophilus* strains isolated from Turkish homemade yoghurts. African Journal of Microbiology Research. 2011;**5**(6):675-682. DOI: 10.5897/ AJMR10.835

*Current Issues and Challenges in the Dairy Industry*

[20] Dixon B. Secrets of the Bulgarian bacillus. The Lancet Infectious Diseases.

[21] Makino S, Ikegami S, Kume A, Horiuchi H, Sasaki H, Orii N. Reducing the risk of infection in the elderly by dietary intake of yoghurt fermented with *Lactobacillus delbrueckii* ssp. *bulgaricus* OLL1073R-1. British Journal of Nutrition. 2010;**104**(7):998-1006. DOI: 10.1017/S000711451000173X

[22] Yamamoto Y, Fujino K, Saruta J, Takahashi T, To M, Fuchida S, et al. Effects of yogurt fermented with *Lactobacillus delbrueckii* ssp. *bulgaricus* OLL1073R-1 on the IgA flow rate of saliva in elderly persons residing in a nursing home: A before-after non-randomised intervention study. Gerodontology. 2017;**34**(4):479-

485. DOI: 10.1111/ger.12296

2002;**79**(1-2):3-16

[23] Ross RP, Morgan S, Hill C. Preservation and fermentation: Past, present and future. International Journal of Food Microbiology.

[24] Cizeikiene D, Juodeikiene G, Paskevicius A, Bartkiene E.

Antimicrobial activity of lactic acid bacteria against pathogenic and spoilage microorganism isolated from food and their control in wheat bread. Food Control. 2013;**31**(2):539-545. DOI: 10.1016/j.foodcont.2012.12.004

[25] Reis JA, Paula AT, Casarotti S, Penna A. Lactic acid bacteria antimicrobial compounds: Characteristics and

applications. Food Engineering Reviews. 2012;**4**. DOI: 10.1007/s12393-012-9051-2

[26] Bachrouri M, Quinto EJ, Mora MT. Kinetic parameters of *Escherichia coli* O157: H7 survival during fermentation of milk and refrigeration of home-made yoghurt. International Dairy Journal.

[27] Akpinar A, Yerlikaya O, Kiliç S. Antimicrobial activity and antibiotic

2006;**16**(5):474-481

2002;**2**(4):260

[14] Kulp WL, Rettger LF. Comparative Study of *Lactobacillus acidophilus* and *Lactobacillus bulgaricus*. Journal of Bacteriology. 1924;**9**(4):357-395 Retrieved from: http://www.ncbi.nlm. nih.gov/pmc/articles/PMC379059/

[15] Lick S, Drescher K, Heller KJ. Survival of *Lactobacillus delbrueckii* subsp. *bulgaricus* and *Streptococcus thermophilus* in the Terminal Ileum of Fistulated Göttingen Minipigs. Applied and Environmental

Microbiology. 2001;**67**(9):4137-4143. DOI: 10.1128/AEM.67.9.4137-4143.2001

[16] Elli M, Callegari ML, Ferrari S, Bessi E, Cattivelli D, Soldi S, et al. Survival of yogurt bacteria in the human gut. Applied and Environmental Microbiology.

[17] Mater DDG, Bretigny L, Firmesse O, Flores M-J, Mogenet A, Bresson J-L, et al. *Streptococcus thermophilus* and *Lactobacillus delbrueckii* subsp. *bulgaricus* survive gastrointestinal transit of healthy volunteers consuming yogurt. FEMS Microbiology Letters. 2005;**250**(2):185-187. DOI: 10.1016/j.

[18] Vodnar DC, Socaciu C, Rotar AM,

Technology. 2010;**45**(11):2345-2351. DOI: 10.1111/j.1365-2621.2010.02406.x

[19] Moro-Garcia MA, Alonso-Arias R, Baltadjieva M, Benitez CF, Barrial MAF, Ruisánchez ED, et al. Oral supplementation with *Lactobacillus delbrueckii* subsp. *bulgaricus* 8481 enhances systemic immunity in elderly subjects. Age. 2013;**35**(4):1311-1326. DOI: 10.1007/s11357-012-9434-6

Stãnilã A. Morphology, FTIR fingerprint and survivability of encapsulated lactic bacteria (*Streptococcus thermophilus* and *Lactobacillus delbrueckii* subsp. *bulgaricus*) in simulated gastric juice and intestinal juice. International Journal of Food Science and

2006;**72**(7):5113-5117

femsle.2005.07.006

**108**

[28] Tufail M, Hussain S, Malik F, Mirza T, Parveen G, Shafaat S, et al. Isolation and evaluation of antibacterial activity of bacteriocin produced by *Lactobacillus bulgaricus* from yogurt. African Journal of Microbiology Research. 2011;**5**(22):3842-3847

[29] Boyanova L, Gergova G, Markovska R, Yordanov D, Mitov I. Bacteriocinlike inhibitory activities of seven *Lactobacillus delbrueckii* subsp. *bulgaricus* strains against antibiotic susceptible and resistant *Helicobacter pylori* strains. Letters in Applied Microbiology. 2017;**65**(6):469-474. DOI: 10.1111/lam.12807

[30] Tongtawee T, Dechsukhum C, Leeanansaksiri W, Kaewpitoon S, Kaewpitoon N, Loyd RA, et al. Improved helicobacter pylori eradication rate of tailored triple therapy by adding *Lactobacillus delbrueckii* and *Streptococcus thermophilus* in northeast region of Thailand: A prospective randomized controlled clinical trial. Gastroenterology Research and Practice. 2015;**2015**:518018. DOI: 10.1155/2015/518018

[31] Fewtrell LJ, Prüss-Üstün A, Landrigan P, Ayuso-Mateos JL. Estimating the global burden of disease of mild mental retardation and cardiovascular diseases from environmental lead exposure. Environmental Research. 2004;**94**(2):120-133. DOI: 10.1016/ S0013-9351(03)00132-4

[32] Li B, Jin D, Yu S, Etareri Evivie S, Muhammad Z, Huo G, et al. In vitro and in vivo evaluation of *Lactobacillus*  *delbrueckii* subsp. *bulgaricus* KLDS1.0207 for the alleviative effect on lead toxicity. Nutrients. 2017. DOI: 10.3390/nu9080845

[33] Alazzeh AY, Ibrahim SA, Song D, Shahbazi A, AbuGhazaleh AA. Carbohydrate and protein sources influence the induction of α- and β-galactosidases in *Lactobacillus reuteri*. Food Chemistry. 2009;**117**(4):654-659

[34] Ibrahim SA, Gyawali R. Lactose intolerance. In: Young P, George H, editors. Milk and Dairy Products in Human Nutrition: Production, Composition and Health. Iowa, IA: John Wiley & Sons; 2013. pp. 246-256

[35] de Vrese M, Stegelmann A, Richter B, Fenselau S, Laue C, Schrezenmeir J. Probiotics—compensation for lactase insufficiency. The American Journal of Clinical Nutrition. 2001;**73**(2):421s-429s. DOI: 10.1093/ ajcn/73.2.421s

[36] Kechagia M, Basoulis D, Konstantopoulou S, Dimitriadi D, Gyftopoulou K, Skarmoutsou N, et al. Health benefits of probiotics: A Review. ISRN Nutrition. 2013;**2013**:481651. DOI: 10.5402/2013/481651

[37] Singh J, Khanna A, Chander B. Antibacterial activity of yogurt starter in cow and buffalo milk. Journal of Food Protection. 1979;**42**(8):664-665

[38] Abdel-bar NM, Harris ND. Inhibitory effect of *Lactobacillus bulgaricus* on psychrotrophic bacteria in associative cultures and in refrigerated foods. Journal of Food Protection. 1984;**47**(1):61-64

[39] Nagai T, Makino S, Ikegami S, Itoh H, Yamada H. Effects of oral administration of yogurt fermented with *Lactobacillus delbrueckii* ssp. *bulgaricus* OLL1073R-1 and its exopolysaccharides against influenza virus infection in mice.

International Immunopharmacology. 2011;**11**(12):2246-2250

[40] Simova E, Beshkova D, Najdenski H, Frengova G, Simov Z, Tsvetkova I. Antimicrobial-producing lactic acid bacteria isolated from traditional Bulgarian milk products: Inhibitory properties and in situ bacteriocinogenic activity. In: Proceedings of the IUFoST, 13th World Congress Food Sci Technol "Food is life". 2006. pp. 17-21

[41] Stamatova I, Kari K, Meurman JH. In vitro evaluation of antimicrobial activity of putative probiotic lactobacilli against oral pathogens. International Journal of Probiotics and Prebiotics. 2007;**2**(4):225

[42] Banerjee P, Merkel GJ, Bhunia AK. *Lactobacillus delbrueckii* ssp. *bulgaricus* B-30892 can inhibit cytotoxic effects and adhesion of pathogenic *Clostridium difficile* to Caco-2 cells. Gut Pathogens. 2009;**1**:8. DOI: 10.1186/1757-4749-1-8

[43] Zhu Y, Xiao L, Shen D, Hao Y. Competition between yogurt probiotics and periodontal pathogens in vitro. Acta Odontologica Scandinavica. 2010;**68**(5):261-268

[44] Fooladi AAI, Forooshai MC, Saffarian P, Mehrab R. Antimicrobial effects of four lactobacilli strains isolated from yoghurt against Escherichia coli O157: H7. Journal of Food Safety. 2014;**34**(2):150-160

[45] Nour I, Fattouh F, El-Adawi H. Antibacterial bioactivity of selected lactic acid bacterial strains against some human pathogenic bacteria. International Journal of Pharmacology. 2015;**11**(5):440-447

[46] Mahmood T, Masud T, Ali S, Sarfraz Abbasi K, Liaquat M. Optimization and partial characterization of bacteriocin produced by *Lactobacillus* 

*bulgaricus-TLBFT06* isolated from Dahi. Pakistan Journal of Pharmaceutical Sciences. 2015;**28**

[47] Pescuma M, Hébert EM, Haertlé T, Chobert J-M, Mozzi F, de Valdez GF. *Lactobacillus delbrueckii* subsp. *bulgaricus* CRL 454 cleaves allergenic peptides of β-lactoglobulin. Food Chemistry. 2015;**170**:407-414

[48] Laiño JE, Hebert EM, de Giori GS, LeBlanc JG. Draft genome sequence of *Lactobacillus delbrueckii* subsp. *bulgaricus* CRL871, a folateproducing strain isolated from a northwestern Argentinian yogurt. Genome Announcements. 2015;**3**(3):e00693-e00615

[49] El-Adawi H, Nour I, Fattouh F, El-Deeb N. Investigation of the antiviral bioactivity of *Lactobacillus bulgaricus* 761N extracellular extract against hepatitis C virus (HCV). International Journal of Pharmaceutics. 2015;**11**:19-26

[50] Tebyanian H, Bakhtiari A, Karami A, Kariminik A. Antimicrobial activity of some *Lactobacillus* species against intestinal pathogenic bacteria. International Letters of Natural Sciences. 2017

[51] Ravindran L, Manjunath N, Darshan RP, Manuel SGA. In vitro study analysis of antimicrobial properties of lactic acid bacteria against pathogens. Journal of Bio Innovation. 2016;**5**(2):262-269

[52] Abedi D, Feizizadeh S, Akbari V, Jafarian-Dehkordi A. In vitro anti-bacterial and anti-adherence effects of *Lactobacillus delbrueckii* subsp *bulgaricus* on *Escherichia coli*. Research in Pharmaceutical Sciences. 2013;**8**(4):260-268. Retrieved from: https://www.ncbi.nlm.nih.gov/ pubmed/24082895

[53] Guha D, Banerjee A, Mukherjee R, Pradhan B, Peneva M, Aleksandrov G,

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et al. A probiotic formulation containing *Lactobacillus bulgaricus* DWT1 inhibits tumor growth by activating pro-inflammatory responses in macrophages. Journal of Functional

Foods. 2019;**56**:232-245

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et al. A probiotic formulation containing *Lactobacillus bulgaricus* DWT1 inhibits tumor growth by activating pro-inflammatory responses in macrophages. Journal of Functional Foods. 2019;**56**:232-245

*Current Issues and Challenges in the Dairy Industry*

*bulgaricus-TLBFT06* isolated from Dahi. Pakistan Journal of Pharmaceutical

[47] Pescuma M, Hébert EM, Haertlé T, Chobert J-M, Mozzi F, de Valdez GF. *Lactobacillus delbrueckii* subsp. *bulgaricus* CRL 454 cleaves allergenic peptides of β-lactoglobulin. Food Chemistry. 2015;**170**:407-414

[48] Laiño JE, Hebert EM, de Giori GS, LeBlanc JG. Draft genome sequence of *Lactobacillus delbrueckii* subsp. *bulgaricus* CRL871, a folateproducing strain isolated from a northwestern Argentinian yogurt. Genome Announcements.

2015;**3**(3):e00693-e00615

2015;**11**:19-26

Sciences. 2017

pubmed/24082895

[49] El-Adawi H, Nour I, Fattouh F, El-Deeb N. Investigation of the antiviral bioactivity of *Lactobacillus bulgaricus* 761N extracellular extract against hepatitis C virus (HCV). International Journal of Pharmaceutics.

[50] Tebyanian H, Bakhtiari A, Karami

[51] Ravindran L, Manjunath N, Darshan RP, Manuel SGA. In vitro study analysis of antimicrobial properties of lactic acid bacteria against pathogens. Journal of Bio Innovation. 2016;**5**(2):262-269

[52] Abedi D, Feizizadeh S, Akbari V, Jafarian-Dehkordi A. In vitro anti-bacterial and anti-adherence effects of *Lactobacillus delbrueckii* subsp *bulgaricus* on *Escherichia coli*. Research in Pharmaceutical Sciences. 2013;**8**(4):260-268. Retrieved from: https://www.ncbi.nlm.nih.gov/

[53] Guha D, Banerjee A, Mukherjee R, Pradhan B, Peneva M, Aleksandrov G,

A, Kariminik A. Antimicrobial activity of some *Lactobacillus* species against intestinal pathogenic bacteria. International Letters of Natural

Sciences. 2015;**28**

International Immunopharmacology.

Najdenski H, Frengova G, Simov Z, Tsvetkova I. Antimicrobial-producing lactic acid bacteria isolated from traditional Bulgarian milk products: Inhibitory properties and in situ bacteriocinogenic activity. In:

Proceedings of the IUFoST, 13th World Congress Food Sci Technol "Food is

[41] Stamatova I, Kari K, Meurman JH. In vitro evaluation of antimicrobial activity of putative probiotic lactobacilli against oral pathogens. International Journal of Probiotics and Prebiotics.

[42] Banerjee P, Merkel GJ, Bhunia AK. *Lactobacillus delbrueckii* ssp. *bulgaricus*

B-30892 can inhibit cytotoxic effects and adhesion of pathogenic *Clostridium difficile* to Caco-2 cells. Gut Pathogens. 2009;**1**:8. DOI:

[43] Zhu Y, Xiao L, Shen D, Hao Y. Competition between yogurt probiotics and periodontal pathogens in vitro. Acta Odontologica Scandinavica.

[44] Fooladi AAI, Forooshai MC, Saffarian P, Mehrab R. Antimicrobial effects of four lactobacilli strains isolated from yoghurt against Escherichia coli O157: H7. Journal of Food Safety. 2014;**34**(2):150-160

[45] Nour I, Fattouh F, El-Adawi H. Antibacterial bioactivity of selected lactic acid bacterial strains against some human pathogenic bacteria. International Journal of Pharmacology.

[46] Mahmood T, Masud T, Ali S, Sarfraz Abbasi K, Liaquat M. Optimization and partial characterization of bacteriocin produced by *Lactobacillus* 

10.1186/1757-4749-1-8

2010;**68**(5):261-268

2015;**11**(5):440-447

2011;**11**(12):2246-2250

life". 2006. pp. 17-21

2007;**2**(4):225

[40] Simova E, Beshkova D,

**110**

[54] Doncheva NI, Antov GP, Softova EB, Nyagolov YP. Experimental and clinical study on the hypolipidemic and antisclerotic effect of *Lactobacillus bulgaricus* strain GB N 1 (48). Nutrition Research. 1 April 2002;**22**(4):393-403

## *Edited by Salam A. Ibrahim, Tahl Zimmerman and Rabin Gyawali*

The dairy industry has faced several challenges that have impacted dairy food quality and consumer acceptability. This book presents a different approach to address current issues and challenges facing the dairy industry. The book consists of seven chapters dealing with dairy processing, current issues related to consumers, and probiotic characteristics. We hope that this first edition can build interest among other scientists to join our future effort to write a more comprehensive book on this topic.

Published in London, UK © 2020 IntechOpen © Dmytro Synelnychenko / iStock

Current Issues and Challenges in the Dairy Industry

Current Issues and Challenges

in the Dairy Industry

*Edited by Salam A. Ibrahim,* 

*Tahl Zimmerman and Rabin Gyawali*