**4. Manufacturing of fermented milks**

Characteristics of naturally fermented milk depend upon the availability of the milk in respective regions. However, fermented milk like Zabady, Laban, Rayeb, and Shubat from Northern Africa (Egypt, Morocco, and mid-west Asian countries) and Shoyu from Himalayan region as well as flmjölk and långfl from Sweden have same characteristics of fermentation [5, 7].

**Figure 1.**

*General steps for manufacturing of bifidus milks.*

In general, manufacturing process of fermented milk is in **Figure 1**. The technology and biochemistry of fermentation have been extensively investigated and varied among countries, regions or for special purposes. However, the principal stages of production of any type of fermented milk are included: standardization of fat content, fortification of the milk solids, homogenization, deaerization (optional), and high heat treatment have much in common reviewed. Fermented dairy products, included probiotics cultures, are generally not commercialized as in single cultures, except Yakult and Bifidus milk or acidophilus milk. However, Fermented milks contain mixed cultures.

## **5. Fermented non-bovine milk and vegetable extract**

Fermented non-bovine milks grew along with fermented cow milk, despite its limited numbers.

Four major ruminant species (cow, goat, sheep, and buffalo) contribute raw milk for their manufacturing, on which the research has been focusing since a long time. Currently, the worldwide production of non-bovine milk reaches 133 million tons per year, accounting for more than 17% of all milk production. Of this amount, 13.5% accounts from goat milk production. Goat milk is one of the major contributors to non-bovine milk production [8]. In recent years, the social transformation underway in poor and developing countries, climatic changes, and increased attention to animal welfare are highlighting on some minor animal species that have a "local dairy concern" such as equines, camels, and yaks [9]. It has studied fermented bovine and non-bovine milk (goat and mare's milk) using raw milk or whey liquid added by probiotic bacteria, either single or multiple probiotic culture [3]. Goat milk possesses beneficial characteristics that make it potentially useful in human medicine or nutrition. Fermented Goat Milk developed in the presence of *L. casei* [10]. As well as present of *Streptococcus thermophilus* or mix probiotic cultures could well accepted by consumers [11].

The so-called vegetable milks are in the spotlight as the beverage that is lactosefree, animal protein-free and cholesterol-free features which fit well with the current demand for healthy food products [12–14]. There is an increasing demand for non-dairy probiotic foods (both fermented and non-fermented) including fruit and vegetable juices, soy and certain cereal products due to vegetarianism [15]. Soy Milk is rich in essential and branched amino acids except Met, and good source of B- vitamins especially, niacin, pyridoxine and folic acid/folacin [13]. Asia represents high demand of vegetable beverages, included fermented vegetable milk as Soymilk [12]. The food market reflects to an ever-greater degree the consumer demand for healthy food products, especially fermented beverage using cereals, nuts as almonds, peanuts, as well as fermented soymilk. France may be a country that considers, it is not milk, but extract [16]. There are two large different groups of vegetable milks or plant milks: nut and cereal milk that can be broadly classified by [12].

### **6. Probiotics and health benefits for human being**

Fermented milks or vegetable extracts offers tremendous potential for promoting health, improving nutrition, and reducing the risks of various diseases. Lactic acid bacteria, both mesophilic and thermophilic or intestinal bacteria may be considered as probiotic cultures which improve health status of the host. Infants, children, adults, and elderly can consume fermented milks for their good taste, health benefit and their general nutritional value, reducing the incidence of malnutrition, lactose intolerance and diarrhea. Those with special medical needs can switch to fermented milks to provide added nutrition, solve intestinal disorders, improve immune function, and optimize gut ecology. Offering general health benefit effects for human indicated the role of probiotic bacteria. Modern production makes these products available to world population, resulting in the consumption of fermented milk products will reach new milestone.

Probiotics beverages belong to fermented milks are known as health promoting foods which have market estimated at USD 49.4 billion in 2018 and projected to grow reaching 69.3 billion in 2023. The increasing use of yeast *Saccharomyces boulardii*, follows a success story of probiotic containing yoghurt in China, Brazil, and India. Japan, which has increasingly aging population, consider health benefits associated with probiotic-fortified foods and beverages. Therefore, Japan may have new paradigm Smart community 5.0 to replace Industrial Revolution 4.0, which dehumanized people and communities. Scientists have considered that the minimum effective dose of a probiotic is approximately 108 –109 cells per mL despite there is no evidence-based consensus on the optimal concentration of bacteria pro-dose [17]. The fresh products, including probiotics, contain live starter culture bacteria, while the extended shelf-life products are in doubts of their content of live microorganisms. The most commonly used bacteria, which are found in various probiotic-dairy products, are Bifidobacteria and Lactobacilli. In addition to their desired health and clinical properties, probiotics must meet several basic requirements for the development of marketable probiotic products. The most important requirements are that probiotic bacteria must survive in sufficient numbers in the product, which their physical and genetic stability must be guaranteed during the storage of the product [18].

In recent years, the proposal of new probiotic-related concepts has gained attention. This is because it has been recognized that some mechanisms and clinical benefits cannot be directly related to the live microorganisms [19]. To survive in milk, they need some supplements that act as a growth promoter. Fermented soy milks can be developed using some probiotic cultures as *L. acidophilus, L. casei* as well as *Bifidobacterium pseudolongum* [14, 16, 20], due to the presence of oligosaccharides, belong to non-digestible starch, as Raffinose and Stachyose [21, 22]. These are prebiotic substances (**Figure 2**), which support the growth of probiotic microorganisms (growth promoter). Prebiotics include fructans, oligosaccharides, arabinooligosaccharides, isomaltooligosaccharides, xylooligosaccharides, resistant starch, lactosucrose, lactobionic acid, galactomannan, psyllium, polyphenols, and

**Figure 2.** *Structure of Raffinose and Stachyose of soybean.*

polyunsaturated fatty acids [23]. Soy flour contains fiber as much as 12 g/ 100 g products, equivalent to 1/5 of 60–80 g/ day recommended to maintain a large bowel flora of 1014 microorganisms, typically for a healthy human colon.

*Lactobacillus casei* and especially Bifidobacterium are among the probiotic cultures that defined by Fuller as a live microbial feed supplement which beneficially affect the host (animal) by improving its intestinal microbial balance. However, this definition of Fuller was improved by [24] as "One product contains specific live microbes at the sufficient numbers and change the microbial balance leading to health effect for the host." Probiotic's definition is living microorganisms, which when ingested in sufficient amounts, beneficially influence the health of the host by improving the composition of intestinal microflora [25]. At present, [2, 26] explained that the most frequently utilized species for promoting the health were:


Many probiotic products on the market contain Lactic Acid Bacteria, the population of which tend to be a relatively small proportion of the total gut microflora. There are more than 400 bacterial species in human flora, but only ten species cohabit at the highest population levels. *Bifidobacterium* sp., is the most popular probiotic [14]. These are strict anaerobic and most of them are very sensitive to contact with atmospheric oxygen. The most abundant population belong to the gram-negative Bacteriodes genus (30% of the dominant human flora) and to the Gram-positive Eubacteria, Bifidobacterium, Ruminococcus and the different Clostridia species. Bifidobacteria (**Figure 3**), are accounting for approximately 1% of total microflora.

Bifidobacteries as a single culture cannot grow easily in dairy milk without supplementations despite it grows well in soymilk [14]. The yoghurt bacteria *L. delbrueckii subsp. bulgaricus* and *S. thermophilus* are not mentioned as probiotic culture (false claimed) because they are not typical forming the intestinal flora of man. Despite it, yoghurt that made from dairy milk or yoghurt-like product from vegetable extract are considered as the best-known food vehicle for probiotics because, beyond its own physicochemical and functional characteristics [2, 14].

#### **Figure 3.**

*Bifidobacterium pseudolongum viewed at scanning electron microscopy (SEM), transmission electron microscopy (TEM) and ordinary microscope (OM).*

*Fermentation of Bovine, Non-Bovine and Vegetable Milk DOI: http://dx.doi.org/10.5772/intechopen.96699*

#### **Figure 4.**

*Acidity (left) and bacterial counts (right) of Bifidobacterium sp. in yoghurt (CM), CM+ yeast extract and Soyoghurt (SE).*

**Figure 5.** *B. pseudolongum in yoghurt (left) or soyoghurt (right).*

Role of probiotic cultures to give positive impact for human health depends on if they are life or not in media, as in **Figures 4** and **5** above [14]. These probiotics microbes require a great number of bacteria that genetically stable since the manufacturing, during storage and expressing their health benefit after consumption especially during transit gastric. Consumers will accept the probiotic products that have a pleasant aroma and taste. It should bear in mind that the development of these bacteria either indirectly to fermented milk or directly to human gut also depends on the type of food ingested by humans. Human, included infant live under rather unnatural conditions lead to unfavorable condition causing illness. We eat a great deal of processed antibacterial substances ranging from vinegar to antibiotics, and in many cases sterile food which many affects our access to and colonization by certain type of bacteria. The healthful effect of probiotic -containing fermented milk is presented supplemented or not by prebiotic substances are in (**Table 3**).

The food in Indonesia is quite different from those in Western countries or Japan. The predominant food ingested by the Indonesians are food from vegetable origins, while in Western countries are from animal origins (meat, eggs, and milk)


#### **Table 3.**

*Health effect of fermented milk's probiotics.*

and in Japan which are from sea food origins. Some of the health and nutritional benefit of probiotics cultures containing LAB or others for human are generally are:

#### **6.1 Enhanced lactose digestion**

Lactose, the main carbohydrate in mammalian milk is fermentable compound, and digested by the intestinal enzyme, lactase, present especially in infants and lactose persistent adults. However, in large percent of the world population (70%) who are lactose non-persistent, lactose escapes digestion in the small bowel and is fermented in the colon with the evolution of gases (CO2, H2, CH4) and the production of osmotically simple sugars. The gas, especially H2 will be Trans located to lungs and can be detected as hydrogen exhalations. Improvement of lactose digestion and avoidance of symptoms of intolerance in lactose mal-absorbers are the most profoundly studied health—relevant effect of fermented milk products, despite are not specifically probiotics effects. The word lactose intolerance is well accepted rather than lactose mal-absorbers, due to no absorption in the proper term of lactose in small intestine. The symptom released due to the absence of or insufficient amount of β-*galactosidase* (*lactase*) in the human small intestine to digest the milk sugar, lactose. The prevalence of primary lactose intolerance is 3–5% in Scandinavia, 17% in Finland and Northern France, 65% in southern France, 20–70% in Italy, 55% in Balkans region, 70–80% in Africa, 80% in Central Africa, 90–100% in Eastern Asia, 30% in Northern India, 70% in southern India and 80% in black American [28]. While secondary form of lactose intolerance may be due to inflammation or functional loss of the mucus of small intestine (enteritis, bacterial or parasitic infection or small bowel syndrome) and by protein-energy malnutrition. It is generally accepted that lactose intolerance symptoms will appear after consuming more than 250 mL a day of whole milk [2]. Fermented milk contains less lactose (**Table 4**).

Lactase deficiency is due to low activity of intestinal lactase (lactase phlorizine hydrolase's in enterocyte of villi). The symptoms associated with lactose intolerance included: bloating, belching, flatulence, cramping and watery stools. Individual with this condition experience gastric distress when they consume fresh, unfermented dairy products. Fermented milk products can especially improve lactose digestion in individual lactose intolerance (**Table 5**).

The explanation will be given that microbial β-*galactosidase* auto digest this lactose and reduce the content of lactose present in dairy products between 20–30% of original lactose [33]. Some developed fermented milk has reduced lactose content around 30% with the range from 14 to 74% in fermented milks. The reduction of lactose


#### *Fermentation of Bovine, Non-Bovine and Vegetable Milk DOI: http://dx.doi.org/10.5772/intechopen.96699*

*a Sugars were detected by HPLC using Column Aminex HPX-87 H at flow rate 0.4 ml/min and mobile phase acetonitrile 30% in 0.005 M H2SO4 (Bio-rad), temp 65 C [2, 28–31].*

#### **Table 4.**

*Reduction of lactose content of some fermented milksa .*


#### **Table 5.**

*Percent lactose, cell counts and lactase activity of fermented milk fed to lactase deficient subjects.*

varies either with raw milk, material, bacterial species used and its combination or the present of growth factors. It is therefore, Fermented milk reduce lactose content and consumers will adapt to different levels of lactose in any commercial fermented milks.

These still alive microbes will implant in the small intestine of humans and continue to digest of lactose lead to better adaptability to the present of such compound or giving improve to lactose tolerance. Yoghurt bacteria, *L. acidophilus*, and *Bifidobacterium* sp. are now well known can survive during gastric transit alive, numerous, and active in the human intestine despite it is not natural inhabitants. These bacteria then deliver their lactose-metabolizing enzyme to its site of action over an extended period. Therefore, probiotic-containing fermented milk could partially explain the better lactose digestion after ingestion of milk.

#### **6.2 Colonization and role anti-pathogenic bacteria**

The antimicrobial or antagonistic activity of probiotic cultures is an important property that people need to understand for using it as health benefit microbes for human beings. Breast-fed infants thrive better than bottle-fed or weaned babies do. This, in part can be associated with a predominant bifid gut flora that inhibits growth of coliforms, enterococci, and clostridia [34]. *Bifidobacterium* sp. account for 92% of the intestinal flora of breastfed infants, but only 20% of bottle-fed or weaned infants. The production of acetic and lactic acid has caused lower pH to inhibit pathogenic bacteria lead for children with high numbers of bifidobacteria to resist some enteric infection very effectively. Regularly supplementation of the infant diet with bifidobacteria can maintain normal intestinal condition. When *B. longum* administered orally to germ-free mice, the bacteria colonize the intestinal tract and reach a concentration of 10 9 –10 10/g intestinal content in 2–3 days. Ability to transit gastric of living probiotic bacteria is important criteria to be used as human health promoting bacteria [35–37]. Translocation of the colonized *B. longum* to the mesenteric lymph node (MLN), liver and kidney occurs between 1–2 weeks after the association caused neither infection nor harmful effect. When *E. coli* O: 111 or O: 157 was administered orally to germfree mice, they will be translocated to various organs occurred and the mice died by endotoxin shock or organ failure. When *B. longum*—mono associated mice challenged *E. coli* O: 157, the intestinal count of strain O: 157 suppressed at a low concentration and no death after 5 weeks. When with *s* O: 111 at a lethal dose, death was avoided [38, 39]. Treatment and prevention of infectious diarrhea are probably the most widely accepted health benefits of probiotic microorganisms. Rotavirus is the most common cause of acute infantile diarrhea in the world. Well-controlled clinical studies have shown that probiotics such as *L. rhamnosus GG, L. reuteri, L. casei, strain Shirota*, and *B. animalis Bb12* can shorten the duration of acute rotavirus diarrhea with the strongest evidence pointing to the effectiveness of *L. rhamnosus* GG and *B. animalis* Bb12 [40].

#### **6.3 Role anti-cancer, anti-hypercholesteremic and anti-virus**

In order to permanently establish the bacterial strain in the host's intestine, the organism must be able to attach to intestinal mucosal cells. The intestinal microflora within a given individual are remarkably stable, although major differences may exist among different persons.

Nevertheless, administration of especially probiotic cultures to either newborn or adult results in certain change in the microbial profiles and metabolic activities of feces. The Fermented Milks and Lactic Acid Beverages Association in Japan have introduced a standard of a minimum of >1 × 107 CFU/ml or CFU/g viable probiotic cells for fresh dairy products [38]. It has suggested a minimum viable number of 106 CFU/ml or grams but recommended 108 CFU/g to compensate the reduction through passage through the gut.

In contrary to the general definition of probiotics, dead probiotics or bacterial' cell components can still combat cancer effectively [41]. In fact, it has been proposed that inactivated or non-viable probiotic cells, as well as their metabolic byproducts (herein referred as "postbiotics" and "paraprobiotics"), also have the ability to provide benefits to the host's health [19].

*Fermentation of Bovine, Non-Bovine and Vegetable Milk DOI: http://dx.doi.org/10.5772/intechopen.96699*

Probiotics are able to conjugate bile acids and to assimilate cholesterol. The feeding of milk formula supplemented with L. acidophilus to infants was shown to result in lower level of serum cholesterol [42]. While culture of *S. thermophilus* assimilated less cholesterol than *L. delbrueckii subs. Bulgaricus*. L. acidophilus assimilated more cholesterol than those S. thermophiles and a commercial yoghurt. Bifidobacteria were also actively assimilated cholesterol. [43] have studied the influence of probiotic yoghurt on serum lipids in women, found that the concentration of total cholesterol, LDL Cholesterol and triacyl glycerol on serum decreased after consuming the standard and this reduction did not depend on the cholesteroemi status (**Table 5**).

However, the beneficial impact of probiotic is not limited to the gut-associated diseases alone, but also in different acute and chronic infectious diseases. This is because probiotics are able to modify the intestinal microbial ecosystem, enhance the gut barrier function, provide competitive adherence to the mucosa and epithelium, produce antimicrobial substances, and modulate the immune activity by enhancing the innate and adaptive immune response. Dairy-based matrix is suitable for proliferation of probiotics due to supplying a rich source of carbon and essential amino acids as a consequence of lactose hydrolyzing and proteolytic systems involved casein utilization [44]. Oral administration of *L. casei* SY13 also significantly enhanced the gut microbial diversity [45]. When, such change even minor applied to pathologic situation, they are sufficient to beneficially alter the course of disease, especially that are associated with the developments of colon cancer [46] as in **Table 6**.

It has explained that some research results using *L. acidophilus* (1 × 109 CFU/ day) and *B. bifidum* (1 × 1010 CFU/day) during 3 weeks reduced fecal activity of

