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Lactobacillus is an important class of Gram-positive, non-spore-forming bacteria for food industrial applications. The genus Lactobacillus is a potential candidate in fermentation technology for the production of fermented food, feed, and pharmaceutical products. The diverse features of Lactobacilli based on their capability to produce acids, enzymes, bacteriocins by fermenting carbohydrates. Lactobacilli have probiotic potential and therefore applied in dairy [cheese, yoghurt, fermented milk] and nondairy products such as sausages, juices as well as in animal feed in the form of starter culture. Among Lactobacilli, lactic acid-producing bacteria are applied as starter cultures in a variety of fermented foods. Lactobacilli are the natural microflora of the gastrointestinal tract and play a beneficial role against infections. The ability of Lactobacilli to produce bacteriocins and other antifungal compound lead to the development of bioprotective cultures for use in different foods. Bacteriocins has wide applications in food industries for preventing the attack of foodborne pathogens and for manufacturing active packaging materials. This chapter aimed to review significant industrial applications of Lactobacilli with specified strains and also starter cultures with their potential beneficial effects are engrossed. The chapter highlights the commercial applications of Lactobacilli in the food, feed, wine and pharmaceutical industries.
Faculty of Agriculture, Department of Food Science and Technology, University of Poonch, Rawalakot, Azad Kashmir, Pakistan
Turfa Mamoona
Faculty of Agriculture, Department of Food Science and Technology, University of Poonch, Rawalakot, Azad Kashmir, Pakistan
Saiqa Bashir
Faculty of Agriculture, Department of Food Science and Technology, University of Poonch, Rawalakot, Azad Kashmir, Pakistan
Imtiaz Hussain
Faculty of Agriculture, Department of Food Science and Technology, University of Poonch, Rawalakot, Azad Kashmir, Pakistan
Imran Hayat
Faculty of Agriculture, Department of Food Science and Technology, University of Poonch, Rawalakot, Azad Kashmir, Pakistan
*Address all correspondence to: nagina_rafiq2008@yahoo.com;, drnaginarafiq@upr.edu.pk
1. Introduction
The genus Lactobacillus was discussed for the first time in 1901. These are rod- shaped, nonspore-forming, Gram-positive bacteria found in fermented foods and are member of the gastro-intestinal tract and vaginal cavity of humans [1, 2, 3, 4]. Lactobacillus is a group of bacteria with diverse species having higher GC content ranging from 32 to 53 mol% [3]. Morpho-microscopically Lactobacilli are rod shaped, non-motile and varying in length ranging from short to long. Lactobacilli are comprised a group of multifunctional microbes with shelf life extending characters of food as well as improving human health in the form of probiotics [4]. Studies of the whole bacterial genomic sequence enabled, the scientists to reclassify the species of Lactobacilli into 25 genera including an edited genus of Lactobacillus i.e. Paralactobacillus and Lactobacillus delbrueckii group and 23 other novel genera containing Acetilactobacillus, Agrilactobacillus, Amylolactobacillus, Apilactobacillus, Bombilactobacillus, Companilactobacillus, Dellaglioa, Fructilactobacillus, Furfurilactobacillus, Holzapfelia, Lacticaseibacillus, Lactiplantibacillus, Lapidilactobacillus, Latilactobacillus, Lentilactobacillus, Levilactobacillus, Ligilactobacillus, Limosilactobacillus, Liquorilactobacillus, Loigolactobacilus, Paucilactobacillus, Schleiferilactobacillus, and Secundilactobacillus [5]. Overall, more than 250 species have been assigned to the genus Lactobacillus previously and are now referred to with a new genus name. Lactobacillus releases lactic acid as the major end product of metabolism during fermentation of glucose with small amounts of acetic and succinic acid [2, 3].
The most common habitat of Lactobacilli is the gastro-intestinal tract of mammals where these bacteria may colonize. This group of bacteria is also commonly found in mouth such as in tooth plaque, saliva, vaginal tract of humans and mammals [2, 3]. Lactobacillus species commonly isolated from GI tract are Lactobacillus acidophilus, L. brevis, L. plantarum, Lactobacillus salivarius and Lactobacillus fermentum. Lactobacillus are significantly required for gastrointestinal health and are not considered as a pathogen in healthy individuals except when having an association with dental caries [6, 7]. Lactobacilli are usually reflected as proactive microbes as they inhibit the growth of pathogenic microorganisms by producing lactic acid and other metabolic compounds [8]. While in immunocompromised patients’ lactobacilli are intricate as pathogens and various reports of AIDS, organ transplant, neutropenia caused by lactobacillus infection has been published [9, 10, 11]. The most common infections caused by Lactobacilli are abscesses, bacteremia, endocarditis, neonatal meningitis, dental caries and chorioamnionitis [5, 12]. The chapter focuses on an exploration of the industrial potential of Lactobacilli for manufacturing a diversity of products. The purpose of this chapter is to gather literature related to the utilization of Lactobacillus species as a microbial candidate for commercial-scale production of valuable products for improving human and animal health by protecting them against pathogenic microflora. The major emphasis is to strive for the attention towards the sustainable application of potent Lactobacillus species for futuristic prosperities of mankind.
Lactobacillus has a long history of use in biotechnology predominantly in manufacturing and conserving food ingredients by the process of fermentation, yoghurt, cheese, kefir, sauerkraut and many other fermented food products productions with the addition of animal feeds manufacture [13]. Further bioprocess technology has developed the specified production aids and procedures for food and feed ingredients [14].
Lactobacilli were the first microbes used by human beings for processing foods and preservation of foods by inhibiting other microbial invasions that can cause food spoilage and ultimately results in foodborne illness [15, 16]. Lactobacillus is a significant genus to recent food and feed technologies, not only because of its cumulative interest in valuable functional properties. The dairy industry and self-care health productions are dynamically engaged in promoting the usage of Lactobacilli in food, [17]. Some traditional fermented foods (Table 1) based on Lactobacillus fermentation from different origins, cultures, civilizations, customs, social relations and sometimes customized with religions were reported by Food and Drug Administration [18, 19]. Because of the characteristic to produce organic acids by using carbohydrates Lactobacilli have extensive application in the food industry [20].
2.1 Lactobacilli as starters in food processing industries
Lactobacilli are mainly used as a starter culture in the contemporary food processing industry. The starter culture may contain only one pure strain or a combination of strains from different species of microbes [20]. Starter cultures can be well-defined as the enormous number of cell preparations, any of a single strain type or an assortment of two or more microbial strain/species that are supplemented in foods to get the advantage of consequent products or compound obtained from their metabolic or enzymatic activity [21]. Since starter cultures are used in food production settings to accomplish processes of fermentation, their usage is globally a common practice in food manufacturing industry. This has led to the commercialization of numerous products such as probiotics or starters, bio-protective cultures that are intended to deliver foods with specific nutritional and sensual characteristics, prospective health benefits and assurance of food safety [22]. Starter cultures are practically applied in a wide array of food industries such as the dairy productions for the manufacture of yoghurt, cheese and other fermented products from dairy milk [23], sausage manufacture in meat industry [24], in wine and beer industry for the production of alcohol [25, 26], production of vinegar [27], preparation of soy and rice-based oriental foodstuffs [28], fermented cereals and bakery products [29], and in making of fermented products from fruits and vegetables [30, 31, 32]. Figure 1 clearly illustrates the industrial potential of lactobacilli.
Figure 1.
Application of lactobacillus in food industry.
Starter cultures may contain any type of microbes such as bacteria, yeast, mold or any type of fungal culture. But the species of Lactobacilli reported in the literature have vast potential for usage as starter culture because of its antimicrobial effects which ultimately prove beneficial for food safety. Potential antimicrobial properties of some lactobacilli have been described by scientists such as Latilactobacillus sakei isolated from meat sausages have an antimicrobial effect against Listeria monocytogenes, Salmonella spp. and E. coli in combination of garlic powder and wine [33]. Another strain of lactobacillus i.e. Lactiplantibacillus plantarum was found effective against pathogenic bacteria of the Enterobacteriaceae family when used as a starter in Vacuum packaged products [34].
2.2 Lactobacilli in dairy industry
The Lactobacillus genus comprise a large number of diverse species with relatively large degree of variation and diversity. It is one of the largest genus in the group of lactic acid bacteria of over fifty species [35]. Lactobacilli are the important lactic acid producing bacteria and are responsible for the development of microflora in the most dairy products particularly cheese and fermented milk. Lactobacillus is the significant microorganism for development of color, flavor and texture of dairy products via acidification owed to lactic acid and by metabolizing milk proteins. The commonly used species of Lactobacilli in the dairy industry are Lb. lactis, Lb. casei, Lb. rhamnosus, Lb. helveticus, Lb. plantarum and Lb. curvatus [36]. Moreover, lactobacilli are incorporated in the preparation of cheese, yoghurt and fermented milk as probiotic cultures because of their potential benefits for the treatment of acute and chronic inflammation of the gastrointestinal tract [37]. Lactobacilli can additionally be used for the preservation of dairy products as they can produce bacteriocins which are usually inhibiting the growth of other non-desired microbes [38]. The species of Lb. delbrueckii encompasses three subspecies that is subsp. bulgaricus, subsp. delbrueckii and subsp. lactis [35] are used as a starter culture for yoghurt preparation. The information related to the use of other cultures for yoghurt manufacturing and safety is scarce.
The strain Lb. paracasei subsp. paracasei is recovered commonly from ripened cheese and forms a constitute with Lb. curvatus, Lb. plantarum, Lb. casei and Lb. rhamnosus is the primary microbiota of the non-starter Lactic acid bacteria group contributing to the process of cheese maturation [39, 40]. Among the species of lactobacilli Lb. rhamnosus is one that has been applied as a probiotic organism in functional foods. The strain Lb. rhamnosus has been identified and designated with a number HN001, has both probiotic and flavor improving qualities, hence, it can be useful as an adjunct in the manufacturing of cheese to decrease dangerous microflora, improve the flavor of cheese and increase the speed of cheese ripening [41, 42]. Lactobacillus johnsonii formerly known as Lb. acidophilus has been widely studied because of its probiotic characteristic and is commercialized for production of fermented milk (LC1) products [43]. The Lb. johnsonii displays antimicrobial [44, 45, 46] and immunomodulatory properties [47, 48] properties. A thermophilic starter culture of Lactobacillus helveticus is used in the production of a numeral fermented products of dairy [49] and develops on a comparatively limited number of carbohydrates such as galactose and lactose with typical requirement of few vitamins for growth [50, 51]. Knowledge about the biodiversity of Lactobacillus can be employed as a starter culture in different dairy products described in previous literature is provided in the tabulated form (Table 2).
Name
Country of origin
Description
Lactobacillus Species
Gundruk
Nepal
Prepared by fermentation of leafy vegetables. It is an appetizer.
Concentrated sour buttermilk used in the manufacturing of Kishk, sometimes used in salad or to make a beverage after dilution with water; semisolid consistency, tart and salty taste
Lb. casei, Lb. plantarum, Lb. brevis
Koumiss
Kazakhstan and Kyrgyzstan
Milk is processed into a drink by lactic acid and alcoholic fermentation
Lb. acidophilus, Lb. bulgaricus, S. thermophilus, Bifidobacterium
Fermented milk drink
Lb. casei Shirota
Fruit yoghurt drink
Lb. casei
Bio pouring yoghurt
Probiotic
Fat-free yoghurt drink
Lb. casei
Natural Greek style yoghurt
Lb. acidophilus, Lb. bulgaricus, S. thermophilus
Goat fruit yoghurt
S. thermophilus, Lb. casei
Natural goat yoghurt
Bifidobacterium longum, Lb. acidophilus
Fruit yoghurt
B. animalis subsp. lactis, Lb. acidophilus
Thick and creamy yoghurt
Bifidobacterium, Lb. acidophilus, S. thermophilus
Natural fresh and mild yoghurt
Lb. acidophilus, B. longum, S. thermophilus
Organic natural yoghurt
Lb. acidophilus, Bifidobacterium
Table 2.
Species of lactobacilli present naturally or added as starter in dairy products.
2.3 Lactobacilli in non-dairy food products of plant and animal origin
The species of Lactobacillus are useful for production of fermented foods other than dairy such as sauerkraut by fermenting cabbage. Usually, normal microflora of cabbage with added Lb. plantarum act as starter and salt like sodium chloride is used for preventing the growth of foodborne pathogenic microorganisms [68]. An additional study reported the use of both Lb. plantarum and Lb. brevis for production of sauerkraut [69]. Another food product in which the genus lactobacillus is involved is pickle where the propagation of starter culture of Lb. plantarum is responsible for achieving the desired effect. Another investigation reported the involvement of Lb. versmoldensis sp. nov.; Lb. plantarum; Lb. casei; Lb. pentosus for the production of fermented green olives [70, 71, 72]. The Lactobacillus species such as Lb. sakei and Lb. curvatus were found in dry sausages [73, 74, 75]. A previous study reported the presence of Lb. sanfranciscensis, Lb. reuteri and Lb. pontis in sourdough [76].
L. acidophilus was reported in chocolate by Rosell [77] and Lb. plantarum was found in juice drinks [78] as well as Lb. rhamnosus was described as a microbe found in juice drinks (Valio Gelfilus) GG (ATCC 53103) [64].
L. fermentum has been used extensively as a probiotic strain and may be found in cereal-based fermented foods and many other vegetables [79]. The predominant bacteria of sourdough and cereal based fermented products is Lb. plantarum and is a leading microbe because it applied for producing corn dextrins after the reduction of fermentable sugars [80] Lactobacillus sanfranciscensis is the leading lactic acid bacteria in sourdough [80] and 14 genome assemblies of Lb. sanfranciscensis have been reported [35].
A fermented beverage tea known as Kombucha is prepared by the traditional fermentation process of fermenting sweetened black tea with tea fungus which contains a consortium of yeasts and acetic acid-producing bacteria. The viability of selected strains of Lactobacilli throughout the fermentation process of Kombucha and their interaction with tea fermenting fungus and their role in obtaining a tea beverage with improved functional properties were tested by scientists [81] Five wild strains (Lactobacillus hilgardii, L. fermentum, three strains of L. plantarum) isolated from conventional fermented foods and were added separately in Kombucha on the second day of fermentation. The addition of wild species of Lactobacillus during fermentation of Kombucha contributed significantly to increasing the lactic acid content of the tea beverage. The strain of Lactobacillus plantarum was reported for the highest lactic acid production during Kombucha fermentation. The L. hilgardii strain isolated from sour dough exhibited the highest sustainability in Kombucha and also has possible probiotic characteristics. So this strain can be used in Kombucha fermentation.
2.4 Lactobacilli in wine industry
The major role of lactic acid bacteria in wine production is to conduct the process of malolactic fermentation. The process of wine fermentation reduces the acidity in wine, enhances the mouth feel and aroma and also improves the microbial stability of wine. Therefore, Lactobacilli have application in the production process of wine from both grapes and fruits cider [82]. During the process of malolactic fermentation malic acid is converted into lactic acid and carbon dioxide which is extensively required for the maturation of fruit wine [83]. The recent efforts focused towards discovering the biodiversity of geographic areas associated for wine production with the purpose of finding new strains of lactic acid bacteria which can be used as starters [84]. For instance, two potent new autochthonous malolactic fermentation starters i.e. Lactobacillus paracasei UVI-2 and Lb. hilgardii UVI-23, have been isolated and identified from Albariño grapes in Val do Salnés, Spain [84]. The following species of Lactobacilli such as Lb. plantarum [85], Lb. brevis; Lb. collinoides; Lb. hilgardii; Lb. paracasei; Lb. pentosus; Lb. plantarum; Lb. mali [86] are naturally present or added as a starter during the fermentation processing of wine. Lactobacillus sakeisubsp. sakei was first isolated from rice alcoholic beverage i.e. sake and it is the product from which its name was taken [35].
Many potential strains of Lactobacilli having probiotic effect are applied in feed production for poultry and other animals for beneficial purposes. There is no case of foodborne illness has been reported by ingestion of feed containing Lactobacilli [12]. In agricultural practice, Lactobacilli have long been applied for the preservation of grass or maize in the form of fodder [87] where these bacteria accelerate the rate of decline in pH while protecting plant carbohydrates through homofermentation and also conserving proteins from plants by reducing proteolysis and by deamination [88, 89, 90, 91]. According to literature the species of LactobacillusLb. casei, Lb. buchneri [92], Lb. paracasei subsp. paracasei [93], Lb. plantarum [94], Lb. buchneri, Lb. fermentum [95] and Lb. diolivorans sp. nov. [96] have been applied in silage production. Probiotic Lactobacilli are used as an alternative to antibiotics in animal feed because of the safety concerns of antibiotics [97]. Probiotic microbes in animal feed should be live and have zootechnical properties to convert feed and increase weight gain [98]. The supplementation of probiotics has been suggested for handling numerous stress conditions or for preventing diseases in several animal species [17]. The studies which have highlighted the value of lactobacilli for various animal species are tabularized herein (Table 3). Probiotic treatment is becoming progressively popular in veterinary medicine, particularly for pets.
Animal species
Lactobacillus species
Benefits
Poultry industry
Lactobacillus johnsonii FI9785
Controls endemic necrotic enteritis due to Clostridium perfringens, reducing economic losses associated with the decrease in antibiotic use [99]
Chicks
Lb. acidophilus
Increases body weight gain, decreases fecal weight [100]
Enhances growth rate, body weight gain, Feed conversion and tends to decrease mortality [104]
Dairy cows
Lb. acidophilus, Lb. buchneri 40,788
Significant increase (16.2%) in milk production Used to treat alfalfa silage. Increases the concentration of acetic acid. When fed to lactating cows, it improved the aerobic stability of the ration and increased milk production [105, 106]
Lambs
Lb. species Lb. acidophilus, S. faecium, Lb.casei, Lb. fermentum and Lb. plantarum
Stimulates feed intake and daily weight gain Reduces fecal shedding of E. coli O157:H7 [107, 108]
Table 3.
Health promoting properties of lactobacilli for animal.
Some feed additives of European origin such as BIACTONs N1 12 for piglets contain Lb. farciminis [109]; PROVITA LEs N1 E1706 for calves & piglets contain Lb. rhamnosus DSM 7133 and Enterococcus faecium DSM 7134 [109]; CECT4 529 N123 feed for laying hens hold Lb. acidophilus [110] and Lb. acidophilus was added to the cats and dogs feed [111]. Other Non-European feed additives such as Silage (PRO-STOREs USA) have Lb. casei; Lb. plantarum; Lb. brevis and other lactic acid bacteria and fungi [112]. HP BIOs a feed for cows in USA comprised of Lb. fermentum; Lb. acidophilus; Lb. plantarum; Lb. casei; and lactic acid bacteria [112]. Another feed additive named E-BIOTICTM equine in USA contained Lb. lactic; Lb. acidophilus; Lb. plantarum and other LAB [113].
The use of Lactobacilli has frequently been associated with the prevention of disease. It has been revealed from the literature that the oral administration of large dosages of lactobacilli is an effective and nontoxic in patients having Crohn’s disease, AIDS and various types of enteritis, and in premature babies, children or the senior adults, all of those are possibly at risk of lactobacillemia [114, 115, 116, 117, 118, 119, 120]. In some cases, enteral administration of Lactobacillus species may be favorable [121] in rats and humans [122]. Lactobacilli may also have positive effects in the oral cavity, inhibiting cariogenic Candida and streptococci species [123]. Tallying to these trials, efficient medicines containing viable Lactobacilli with no side effects have developed (Table 4).
Lactobacillus species
Trademark
Dose (Adult) – Comments
Lb. casei rhamnosus
Bacilors (347961.2)
Ingestion, 2 to 8 capsules/day, anti-diarrhea
Lb. acidophilus
Lacteols (305665.6)
Ingestion, 5 tablets/3 to 5 times/day, anti-diarrhea
Lactobacilli can produce lactic acid in large amounts by a biotechnological process through fermentation. Among lactobacilli, lactic acid-producing bacteria play an important role in the production of fermented dairy products and the enzyme beta-galactosidase. Because these bacterial species are generally recognized as safe so enzyme purification does not require any extensive purification processes and also there is no damage to fermented food products [124].
A large number of Gram-negative and Gram-positive bacteria produce antimicrobial protein structures during their growth (either polypeptides or proteins) called bacteriocins [125]. Now a days Lactic acid bacteria have gained specific consideration among the Gram-positive bacterial strains because of the production of bacteriocins [126]. The bacteriocins can be functional in the food industry by employing as natural preservatives. The application of produced bacteriocins having antimicrobial compounds as a natural barrier against food spoilage causing pathogenic microbes has been confirmed to be effective [38]. The following approaches of bacteriocin application in food are usually adopted for the purpose of biopreservation: adding semi purified or pure bacteriocin as additive in food; food is inoculated with the bacteriocin-producing strain; and practice of previously fermented product with a strain having ability to produce bacteriocin as an ingredient in food processing [38]. A new application of bacteriocins is in bioactive packaging which is a procedure that can defend the food from exterior contaminants [126].
Lactobacilli having probiotic effects in humans are completely documented and are dose and strain dependent [127]. Numerous articles, reviews and book chapters have documented the potential lactobacilli to promote overall health: help to mitigate lactose intolerance [128], positively affect the flora of intestinal tract [129], prevent gastrointestinal infections [130, 131], stimulate the immune system [132], decrease in allergies or inflammation responses [133, 134], regulate the motility of gut [135], and promotion of sensation of well-being [136]. Particular health effects such as colon cancer prevention of [137, 138, 139] and decreases in blood lipid concentration and heart disease [140] antihypertensive effects have also been reported. Figure 2 shows the use of probiotics in different food items.
Figure 2.
Probiotic foods in which lactobacilli are applied as a probiotic strain.
6. Contribution of lactobacilli to the United Nations development goals
Lactobacilli in the form of starter culture have prospective potential for application in foods of animal origin. The starter cultures are extensively used in in the meat industry for production of sausage, the dairy industry for yoghurt, cheese and other fermented dairy products and in the fishery industry for fermented fish products. The starter cultures are mainly used in food of animal origin for quick lactic acid production which causes a decrease in pH and thus preventing the growth of spoilage and foodborne pathogenic microbes which ultimately increasing the shelf-life of fermented food products. Also, release of other metabolic compounds (e.g., acetic acid, lactic acid, benzoic acid, propionic acid and hydrogen peroxide or bacteriocins) helps in improving the safety of foods [141]. Since starter cultures have developed as the predominant microbiota of fermented foods and allows the food processors to control the process of fermentation by eliminating the objectionable microflora and reducing the manufacturing and hygienic risks. Another important role of starter cultures is the chemical safety of fermented products/foods by decreasing aromatic polycyclic hydrocarbons and biogenic amine contents. Therefore, the futuristic application of lactobacilli and other microbes as starter culture crucially helps in reducing hunger by elimination of food spoilage which is the main cause of food insecurity and play a significant role in improving health and wellbeing by providing essential nutrients and reducing the population of undesirable microbiota and increasing the shelf life of foods. So the lactobacillus with other microbes can significantly effective for their progressive utilization in important areas of sustainable development goals agenda of 2030, the number one is huger and second is health and wellbeing. At the same time lactobacilli have huge industrial potential and it is the dire need of the day to explore more potential lactobacilli with multiple characteristics for food and pharmaceutical industry.
Lactobacilli, a potentially significant group of microorganisms that play a fundamental role in the fermentation industry for the production of numerous valuable products such as cheese, yoghurt, fermented milk, kefir, enzymes, medicines, etc. Because of the diverse applications lactobacillus genus open new doors for exciting research areas. The application of lactobacilli in feed, pharmaceuticals and probiotics ensures to reduce the rate of mortality and morbidity associated with gastroenteritis and viral infections. The extensive industrial application of Lactobacilli demanded further research to enhance the potential of the species of this group and to isolate efficient strains which can produce multiple products of commercial practice. Lactobacilli have the futuristic potential for utilization in a diversity of industrial applications and hence can be considered a significant candidate for poverty elimination, reducing hunger by producing a diverse range of food products, increasing the shelf life by preserving the food and providing valuable pharmaceutical products.
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Written By
Nagina Rafique, Turfa Mamoona, Saiqa Bashir, Imtiaz Hussain and Imran Hayat
Submitted: 05 February 2022Reviewed: 28 July 2022Published: 17 September 2022
Open access peer-reviewed chapter
