**Author details**

Francesca Patrignani\*, Lorenzo Siroli, Diana I. Serrazanetti and Rosalba Lanciotti

\*Address all correspondence to: francesca.patrignani@unibo.it

Department of Agricultural and Food Sciences, University of Bologna, Bologna, Italy

## **References**


[4] Siroli L, Patrignani F, Serrazanetti DI, Parolin C, Palomino RAN, Vitali B, Lanciotti R. Determination of antibacterial and technological properties of vaginal Lactobacilli for their potential application in dairy products. Frontiers in Microbiology. 2017;**8**:166. DOI: 10.3389/fmicb.2017.00166

regard, dairy products produced with functional lactic acid bacteria strains used as co-starters have been recognized as functional foods able to provide potential benefits in preventing some diseases. Thus, in this scenario the formulation of a soft cheese containing *Lb. crispatus* BC4, isolated from the vagina of healthy women and selected for its antimicrobial activity and its ability to survive during the refrigerated storage, has been assessed in order to develop a new functional soft cheese able to promote the woman's well-being. The results obtained showed that the strain could survive very well in cheese during its refrigerated storage and also survive the simulated stomach duodenum passage maintaining cell loads higher than 6 log cfu/g over the storage. The ability of this strain to interact with the gut microbial population was also studied by using a dynamic model (called SHIME) of the gastrointestinal tract to study physicochemical, enzymatic and microbial parameters, in a controlled in vitro setting,

The results presented in this chapter have highlighted that the innovation in dairy field is achievable using different strategies. The field of high-pressure homogenization certainly represents one of the most important technological tools to reach this aim due to the tangible effects of this non-thermal approach. Also the use of safe and well-characterized probiotic/ health-promoting strains can contribute to the development of products able to increase the general human well-being. Also the interaction between these two strategies could represent

Francesca Patrignani\*, Lorenzo Siroli, Diana I. Serrazanetti and Rosalba Lanciotti

Department of Agricultural and Food Sciences, University of Bologna, Bologna, Italy

[1] Linares DM, Gómez C, Renes E, Fresno JM, Tornadijo ME, Ross RP, Stanton C. Lactic Acid Bacteria and Bifidobacteria with potential to design natural biofunctional health-promoting dairy foods. Frontiers in Microbiology. 2017;**8**:846. DOI: 0.3389/fmicb.2017.00846

[2] Siro I, Kápolna E, Kápolna B, Lugasi A. Functional food product development, marketing and consumer acceptance: A review. Appetite. 2008;**51**:456-467. DOI: 10.1016/j.

[3] Patrignani F, Lanciotti R, Guerzoni ME. Emerging technologies for probiotic and prebi-

otic food. In: Probiotic and Prebiotic Food 2011. ISBN 978-1-61668-842-4

able to affect the viability of *Lb. crispatus* BC4.

46 Technological Approaches for Novel Applications in Dairy Processing

a challenge in the future for the sector's innovation.

\*Address all correspondence to: francesca.patrignani@unibo.it

**2.6. Conclusions**

**Author details**

**References**

appet.2008.0


[14] Senorans FJ, Ibanez E, Cifuentes A. New trends in food processing. Critical Reviews in Food Science and Nutrition. 2003;**43**:507-256

[27] Burns P, Patrignani F, Tabanelli G, Vinderola G, Siroli L, Reinheimer J, et al. Potential of high pressure homogenisation on probiotic Caciotta cheese quality and functionality.

Potential of High-Pressure Homogenization and Functional Strains for the Development…

http://dx.doi.org/10.5772/intechopen.74448

49

[28] Iordache M, Jelen P. High pressure microfluidization treatment of heat denatured whey proteins for improved functionality. Innovative Food Science and Emerging

[29] Bury D, Jelen P, Kaláb M. Disruption of *Lactobacillus delbrueckii* ssp. *bulgaricus* 11842 cells for lactose hydrolysis in dairy products: A comparison of sonication, high-pressure homogenization and bead milling. Innovative Food Science and Emerging Technologies.

[30] Roesch R, Corredig M. Texture and microstructure of emulsions prepared with soy protein concentrate by high-pressure homogenization. LWT—Food Science and Technology.

[31] Saito M, Lin Y, Kobayashi I, Nakajima M. Preparation characteristics of oil-in-water emulsions stabilized by proteins in straight-through microchannel emulsification. Food

[32] Lanciotti R, Patrignani F, Iucci L, Saracino P, Guerzoni ME. Potential of high pressure homogenization in the control and enhancement of proteolytic and fermentative activities of some Lactobacillus species. Food Chemistry. 2007;**102**:542-550. DOI: 10.1016/j.

[33] Patrignani F, Iucci L, Lanciotti R, Vallicelli M, Maina Mathara J, Holzapfel WH, et al. Effect of high pressure homogenization, not fat milk solids and milkfat on the technological performances of a functional strain for the production of probiotic fermented

[34] Gomes AMP, Malcata FX, FAM K, Grande HJ. Incorporation and survival of Bifidobacterium sp. strain Boand lactobacillus acidophilus strain Ki in a cheese product. Netherlands

[35] Vinderola CG, Bailo N, Reinheimer JA. Survival of probiotic microflora in Argentinean yogurts during refrigerated storage. Food Research International. 2000;**33**:97-102 [36] Kasımoglu A, Goncuoglu M, Akgu S. Probiotic white cheese with lactobacillus acidophi-

[37] Menendez S, Centeno JA, Godınez R, Rodrıguez-Otero JL. Effect of Lactobacillus strains on the ripening and organoleptic characteristics of Arzua´–Ulloa cheese. International

[38] Buriti FCA, da Rocha JS, Saad SMI. Incorporation of Lactobacillus acidophilus in minas fresh cheese and its implications for textural and sensorial properties during storage.

Journal of Functional Foods. 2015;**13**:126-136. DOI: 10.1016/j.jff.2014.12.037

Technologies. 2003;**4**:367-376

Hydrocolloids. 2005;**19**:745-751

milks. Journal of Dairy Science. 2007;**90**:4513-4523

lus. International Dairy Journal. 2004;**14**:1067-1073

Journal of Food Microbiology. 2000;**59**:37-46

International Dairy Journal. 2005;**15**:1279-1288

Milk and Dairy Journal. 1995;**49**:71-95

foodchem.2006.06.043

2001;**2**:23-29

2003;**36**:13-124


[27] Burns P, Patrignani F, Tabanelli G, Vinderola G, Siroli L, Reinheimer J, et al. Potential of high pressure homogenisation on probiotic Caciotta cheese quality and functionality. Journal of Functional Foods. 2015;**13**:126-136. DOI: 10.1016/j.jff.2014.12.037

[14] Senorans FJ, Ibanez E, Cifuentes A. New trends in food processing. Critical Reviews in

[15] Wan J, Mawson R, Ashokkumar M, Ronacher K, Coventry MJ, Roginski H, Versteeg C. Emerging processing technologies for functional foods. The Australian Journal of

[16] Wan J, Conventry J, Sanguansri P, Versteeg C. Advances in innovative technologies for microbial inactivation and enhancement of food safety—Pulsed electric field and low-

[17] Patrignani F, Lanciotti R. Applications of high and ultra high pressure homogenization for food safety. Frontiers in Microbiology. 2016;**7**:article 1132. DOI: 10.3389/

[18] Zamora A, Guamis B. Opportunities for ultra-high-pressure homogenisation (UHPH) for the food industry. Food Engineering Reviews. 2015;**7**:130-142. DOI: 10.1007/

[19] Kananen A, Savolainen J, Mäkinen J, Perttilä U, Myllykoski L, Pihlanto-Leppälä A. Influence of chemical modification of whey protein conformation on hydrolysis with

[20] Floury J, Bellettre J, Legrand J, Desrumaux A. Analysis of a new type of homogenizer. A study of the flown pattern. Chemical Engineering & Science. 2004;**59**:843-853

[21] Guerzoni ME, Lanciotti R, Westall F, Pittia P. Interrelation between chemicophysical variables, microstructure and growth of listeria monocytogenes and Yarrowia lipolytica

[22] Lanciotti R, Chaves-Lopez C, Patrignani F, Papparella A, Guerzoni ME, Serio A, et al. Effects of milk treatment with dynamic high pressure on microbial populations, and lipolytic and proteolytic profiles of Crescenza cheese. International Journal of Dairy

[23] Lanciotti R, Vannini L, Pittia P, Guerzoni ME. Suitability of high dynamic-pressuretreated milk for the production of yogurt. Food Microbiology. 2004;**21**:753-760

[24] Patrignani F, Serrazanetti DI, Mathara JM, Siroli L, Gardini F, Holzapfel WH, et al. Use of homogenisation pressure to improve quality and functionality of probiotic fermented milks containing Lactobacillus rhamnosus BFE 5264. International Journal of Dairy

[25] Hayes MG, Kelly AL. High pressure homogenisation of raw whole bovine milk (a) effects on fat globules size and other properties. Journal of Dairy Research. 2003;**70**:297-305 [26] Vannini L, Lanciotti R, Baldi D, Guerzoni ME. Interactions between high pressure homogenization and antimicrobial activity of lysozyme and lactoperoxidase. Journal of

pepsin and trypsin. International Dairy Journal. 2000;**10**:691-697

in food model systems. Science des les Aliments. 1997;**17**:507-522

temperature plasma. Trends in Food Science and Technology. 2009;**20**:414-424

Food Science and Nutrition. 2003;**43**:507-256

48 Technological Approaches for Novel Applications in Dairy Processing

Dairy Technology. 2005;**60**:167-169

fmicb.2016.01132

s12393-014-9097-4

Technology. 2004;**57**:19-25

Technology. 2016;**69**:262-271

Food Microbiology. 2004;**94**:123-135


[39] Ong L, Henriksson A, Shah NP. Chemical analysis and sensory evaluation of Cheddar cheese produced with Lactobacillus acidophilus, Lb. casei, Lb. paracasei or Bifidobacterium sp. International Dairy Journal. 2007;**17**:937-945

**Chapter 4**

**Provisional chapter**

**Sonocrystallization of Lactose from Whey**

Sukhvir Kaur Bhangu, Muthupandian Ashokkumar

principles of lactose crystallization and sonocrystallization.

**Keywords:** lactose, whey, crystallization, sonocrystallization

**Sonocrystallization of Lactose from Whey**

DOI: 10.5772/intechopen.74759

Whey is a by-product obtained from the cheese-making industry. This by-product is the primary source of high-value products such as whey protein concentrates and lactose. The partial removal of water from the whey is the first step in the recovery of lactose. Then, lactose in the concentrated whey is forced to crystallize through a cooling stage. This conventional process of crystallization is very slow up to 72 h accompanied by the generation of a mixture of lactose types (α, β, and amorphous) and low yield of lactose. These issues have been addressed through the seeding of lactose, the antisolvent crystallization, and more recently, by the crystallization of lactose assisted with low-frequency power ultrasound. Sonocrystallization is known to have a number of specific features that include the enhancement of the primary and secondary nucleation, as well as the development of smaller crystals with more uniform sizes and higher purity. Nowadays, there are a number of studies that provide relevant information on the effects of ultrasound on lactose crystallization, although some of these effects are still not fully understood. This book chapter discusses the current knowledge on lactose sonocrystallization and describes the basic

> © 2016 The Author(s). Licensee InTech. 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.

© 2018 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.

The composition of whey varies according to the cheese process, but in general, this contains 0.8–1.0% soluble proteins, 0.05–0.5% fat, >1% salts, 5–6% lactose, and up to 93% of water [1–4]. The volume of whey recovered from a cheese process makes up 70–90% of the original

Yanira Ivonne Sánchez-García,

and Néstor Gutiérrez-Méndez

Yanira Ivonne Sánchez-García,

Muthupandian Ashokkumar and

http://dx.doi.org/10.5772/intechopen.74759

Additional information is available at the end of the chapter

Additional information is available at the end of the chapter

Sukhvir Kaur Bhangu,

**Abstract**

**1. Introduction**

Néstor Gutiérrez-Méndez


#### **Sonocrystallization of Lactose from Whey Sonocrystallization of Lactose from Whey**

DOI: 10.5772/intechopen.74759

Yanira Ivonne Sánchez-García, Sukhvir Kaur Bhangu, Muthupandian Ashokkumar and Néstor Gutiérrez-Méndez Yanira Ivonne Sánchez-García, Sukhvir Kaur Bhangu, Muthupandian Ashokkumar and Néstor Gutiérrez-Méndez Additional information is available at the end of the chapter

Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/intechopen.74759

#### **Abstract**

[39] Ong L, Henriksson A, Shah NP. Chemical analysis and sensory evaluation of Cheddar cheese produced with Lactobacillus acidophilus, Lb. casei, Lb. paracasei or

[40] Blanchette L, Roy D, Belanger G, Gauthier SF. Production of cottage cheese using dress-

[41] Gobbetti M, Corsetti A, Smacchi E, Zocchetti A, De Angelis M. Production of Crescenza cheese by incorporation of bifidobacteria. Journal of Dairy Science. 1998;**81**:37-47

[42] Marino M, Masella R, Bulzomi P, Campesi I, Malorni W, Franconi F. Nutrition and human health from a sex-gender perspective. Molecular Aspects of Medicine. 2011;**32**:

Bifidobacterium sp. International Dairy Journal. 2007;**17**:937-945

1-70. DOI: 10.1016/j.mam.2011.02.001

50 Technological Approaches for Novel Applications in Dairy Processing

ing fermented by bifidobacteria. Journal of Dairy Science. 1996;**79**:8-15

Whey is a by-product obtained from the cheese-making industry. This by-product is the primary source of high-value products such as whey protein concentrates and lactose. The partial removal of water from the whey is the first step in the recovery of lactose. Then, lactose in the concentrated whey is forced to crystallize through a cooling stage. This conventional process of crystallization is very slow up to 72 h accompanied by the generation of a mixture of lactose types (α, β, and amorphous) and low yield of lactose. These issues have been addressed through the seeding of lactose, the antisolvent crystallization, and more recently, by the crystallization of lactose assisted with low-frequency power ultrasound. Sonocrystallization is known to have a number of specific features that include the enhancement of the primary and secondary nucleation, as well as the development of smaller crystals with more uniform sizes and higher purity. Nowadays, there are a number of studies that provide relevant information on the effects of ultrasound on lactose crystallization, although some of these effects are still not fully understood. This book chapter discusses the current knowledge on lactose sonocrystallization and describes the basic principles of lactose crystallization and sonocrystallization.

**Keywords:** lactose, whey, crystallization, sonocrystallization

#### **1. Introduction**

The composition of whey varies according to the cheese process, but in general, this contains 0.8–1.0% soluble proteins, 0.05–0.5% fat, >1% salts, 5–6% lactose, and up to 93% of water [1–4]. The volume of whey recovered from a cheese process makes up 70–90% of the original

© 2016 The Author(s). Licensee InTech. 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. © 2018 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.

volume of milk [3]. Therefore, it is estimated that more than 80 million tons of whey are produced annually all over the world [4, 5].

α isomer will crystallize first in a supersaturated solution of lactose, like a whey concentrate. In this section, the three main phases of lactose crystallization are described: supersaturation,

Sonocrystallization of Lactose from Whey http://dx.doi.org/10.5772/intechopen.74759 53

Supersaturation of lactose solutions is the first step in the crystallization process, since a nonequilibrium condition is required for the spontaneous birth of nuclei [16]. At any given temperature, a maximum amount of solute can be dissolved in a solvent. When a solution is saturated with a solute, this is considered being in a thermodynamic equilibrium. Any further increase in the concentration above the saturation (solubility) point disturbs the equilibrium and induces a pseudo-equilibrium state or supersaturation. The nucleation and hence crystallization won't occur at the supersaturation point (at least not spontaneously), since the energy available is insufficient to induce the nuclei formation. However, beyond the pseudo-equilibrium state (labile zone), nucleation takes place spontaneously. The region between solubility and supersolubility (supersaturation) is known as the metastable zone (MZ). The width of this region (MZW) is obtained by plotting the solubility and supersolubility of the solute as a function of temperature. From these curves, it is possible to establish the temperature and solute concentration required in a crystallization process [5, 17]. The conventional process of lactose crystallization has a wide MZW, which means that a very high supersaturation is

Nucleation has a major influence on crystallization and consequently on the quality properties of lactose crystals like its structure and size distribution [21]. The formation of a new solid phase from a supersaturated solution is called nucleation, and the nucleation rate is the change in the number of particles in solution with time [22]. There are two kinds of nucleations: the primary and secondary; the former occurs when a crystal is nucleated without an interphase in the solution. Nucleation in the absence of solid surfaces is called homogeneous nucleation, and if there is a foreign interphase in the solution, the process is referred as heterogeneous nucleation. In contrast, the secondary nucleation is induced by pre-existing crystals [13]. Two theories try to explain the nucleation mechanism, the Classical Nucleation Theory (CNT) and the Two-Step Nucleation Theory (TSNT). The basics of the CNT are that from a supersaturated solution, a number of ordered subcritical clusters of solute molecules are formed under certain temperature and concentration conditions. When the number of molecules in the cluster increases (until reaching a critical cluster size *n\**) (∼100 to 1000 atoms), the total free energy (*ΔG*) in the system rises. Above this *n\**, the total free energy decreases continuously and the formation of a crystal nuclei becomes favorable. However, a cluster of size *n\** has equal possibilities to form a crystal nucleus or to disaggregate. Therefore, the height of the free energy barrier for nucleation (*ΔG\**) and the nucleation rate are determined largely by the *n\** [16, 21]. The CNT gives some insights about the *n\** and nucleation rate but does not provide information on the structure of aggregates or pathways leading to the formation of solid crystal from the solution [16]. On the other hand, the major difference between the CNT and the TSNT is

nucleation (appearance of crystals), and crystal growth [15].

**2.1. Supersaturation**

necessary to induce nucleation [18, 19].

**2.2. Nucleation**

Most of the small-scale dairy companies dispose of the whey into the municipal sewage, rivers, lakes, or use this by-product as fertilizer and animal feed [4, 5]. The disposal of cheese whey into water bodies and lands should be strongly discouraged because it produces serious environmental problems. The bacterial degradation of whey causes a depletion of oxygen in the water and soil killing aerobic organisms, such as fish, insects, plants, and microorganisms. The high biological and chemical oxygen demand (BOD: 30–50 g L−1; COD: 60–80 g L−1) of the whey arise from its large content of carbohydrates, chiefly lactose (5–6%) [2, 6–8]. In consequence, the removal of lactose reduces more than 80% of the BOD and COD of whey, which minimizes the negative environmental impact of this by-product [9, 10].

Besides the ecological benefits of lactose removal from whey, this by-product also has a great relevance for the food and pharmaceutical industries [11]. It is estimated that 400,000 tons of crystalline lactose are worldwide produced each year. In comparison with other carbohydrates, lactose has a low caloric value, low glycemic index, good plasticity, compressibility, and low level of sweetness. This sugar is used in the food industries in a wide variety of products such as instant coffee, infant formula, and baked foods. Meanwhile, lactose is used as an excipient for tablets and dry powder inhalers in the pharmaceutical industry [11, 12]. The general steps in the recovery of lactose from the whey involve a step for the partial removal of water followed by a crystallization step. Some of the challenges to overcome in the recovery of lactose from the whey are the long crystallization times, low yields, and low quality of lactose crystals. These problems on lactose crystallization have been approached through the seeding of lactose, the use of antisolvent, and more recently, by the sonocrystallization of lactose [1, 3, 5]. In the last years, the number of research studies of the crystallization of lactose assisted with ultrasound has increased considerably. Hitherto, it has been established that sonocrystallization decreases the size of crystals and improves the crystal size distribution but also might speed up the crystallization process or enhance the purity of lactose crystals. However, the effect that ultrasound has on lactose crystallization is by far not fully understood. This chapter discusses the current knowledge on lactose sonocrystallization (fifth section) but also addresses the basic principles of lactose crystallization (second section) and sonocrystallization (fourth section). Furthermore, the conventional process of lactose recovery from whey is described in the third section of this chapter.
