Microbial Safety of Nonalcoholic Beverages

MICROBIAL SAFETY OF NONALCOHOLIC BEVERAGES

6

C.S. Ranadheera⁎, P.H.P. Prasanna†, T.C. Pimentel‡, D.R.P. Azeredo§, R.S. Rocha§, A.G. Cruz§, J.K. Vidanarachchi¶, N. Naumovski‖, R. McConchie#, S. Ajlouni⁎ *

School of Agriculture & Food, Faculty of Veterinary & Agricultural Sciences, The University of Melbourne, Melbourne, VIC, Australia, †Department of Animal & Food Sciences, Faculty of Agriculture, Rajarata University of Sri Lanka, Puliyankulama, Sri Lanka, ‡Federal Institute of Paraná, Paranavaí, Brazil, §Department of Food, Federal Institute of Rio De Janeiro (IFRJ), Rio De Janeiro, Brazil, ¶Department of Animal Science, Faculty of Agriculture, University of Peradeniya, Peradeniya, Sri Lanka, ‖Discipline of Nutrition and Dietetics, Faculty of Health, University of Canberra, Bruce, ACT, Australia, # School of Life and Environmental Sciences, University of Sydney, NSW, Australia

6.1 Introduction Nonalcoholic beverages could vary from hot drinks, dairy-based drinks to soft drinks, and are usually consumed as refreshing drinks or meal replacement. Some examples of popular nonalcoholic beverages are dairy drinks, fruit and vegetable juices, concentrates, soft drinks, energy and sports drinks, hot beverages (tea and coffee), infant formulas, and baby beverages. There are many other miscellaneous nonalcoholic beverage products as well. Most of these nonalcoholic beverages play a significant role in the human diet worldwide due to the high popularity and abundant usage. The nonalcoholic beverage industry is among one of the most dynamic industries in the world (Chavan et  al., 2015; Ranadheera et al., 2017a). These nonalcoholic beverages are competitive and high-performance products in the market, with many launches, presenting various types of packaging and a variety of options. These options for nonalcoholic drinks seem to be limitless nowadays with novel and pleasant flavors and new brands entering the market regularly. Similarly, some old and nonalcoholic beverage products are disappearing from the market regularly due to the better competitiveness among the rising numbers of new products and their good sensory characteristics. Safety Issues in Beverage Production. https://doi.org/10.1016/B978-0-12-816679-6.00006-1 © 2020 Elsevier Inc. All rights reserved.

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The shifting lifestyles of consumers, the growing per capita income, the changing preferences of consumers, and the unique product characteristics such as numerous flavors and variants of beverages have been positively influenced the global nonalcoholic beverage market and will bring in new growth opportunities in the coming decade. In addition, the rising awareness on health benefits of certain nonalcoholic beverage products is amongst the prime factors impeding the development of the global nonalcoholic beverage market (Gawkowski and Chikindas, 2012; Transparency Market Research, 2015). The global nonalcoholic drinks market stood at US$1435.25 billion in 2013 and is anticipated to reach US$1937.73 billion by 2020, expanding at a 4.30% compound annual growth rate (CAGR) from 2014 to 2020. Volume-wise, the nonalcoholic beverage market is poised to rise at a 4.90% CAGR in the forecast period and will fuel the consumption of these products to 1289.03 billion liters by 2020, starting from 912.77 billion liters in 2013 (Transparency Market Research, 2015). Chemical and physical contaminants and microbiological hazards can be considered as the major challenges in the production of safe food and beverage products. At present, there is a considerable burden of foodborne illness, in which pathogenic microorganisms associated with food products and manufacturing processes play a significant role. The microorganisms enter the food chain at different steps are highly versatile and can adapt to the environment allowing survival, growth, and possible production of toxic compounds. Consequently, this cycle of microbial contamination can cause serious illness in humans and significant economic losses to the food and beverage industry (Havelaar et  al., 2010; Ranadheera et  al., 2017c). Due to the procedures involved in manufacturing, product diversity, efficient distribution, abundance, easy accessibility, and extensive consumption, nonalcoholic beverages remain a prevalent vehicle of foodborne diseases. The pathogenic microorganisms from environmental, animal, or human sources can easily enter into beverage food systems during any of the steps from farm to consumer. Nutritionally rich beverages are ideal sources for the growth and survival of pathogenic microorganisms, and can potentially enhance the production of toxic compounds causing foodborne diseases (Ranadheera et  al., 2017c). In order to minimize the microbial cross contamination in beverage production, suitable measures should be taken from the beginning of harvesting and raw material production to beverage manufacturing process, storage, transportation, retailing, and household handling. While the global demands of microbial food safety increase steadily, new threats also continue to be acknowledged frequently. Thus, employing effective and achievable strategies for minimizing or preventing pathogenic microbial contamination of food and beverages are essential. Hence, the new technological developments such

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as advancement in novel tracking and tracing methods for pathogens in beverages are crucial. Due to the complexity of the global nonalcoholic beverage systems, enhancement of improved communications between all parties involved is of the upmost importance: scientists, risk assessors and risk managers, public health authorities, veterinary professionals and food safety experts, as well as consumers (Havelaar et al., 2010; Newell et al., 2010) could also be useful in addressing the microbial food safety issues in nonalcoholic beverages effectively.

6.2  Classification of Nonalcoholic Beverages In general, the global nonalcoholic beverage market could be divided into three broad categories: hot drinks, dairy-based drinks, and soft drinks. Hot drinks include tea, coffee, and hot chocolate-based drinks, whereas soft drinks can be categorized into different groups which include bottled water, carbonated drinks, fruit juices, sports drinks, and other noncarbonated drinks. Examples of dairy drinks are milk, kefir, and Yakult (Fig. 6.1).

6.2.1  Dairy-Based Beverages Most of the dairy beverages are considered as functional foods since they have been shown to be effective as carriers for bioactive ingredients including probiotics, plant sterols, omega-3 fatty acids, vitamins, minerals, and bioactive peptides (Crowley et  al., 2015; Sawale et al., 2016). Among these products, probiotic-based dairy beverages are highly popular and can be considered as one of the widely available beverages in the world at present. Probiotics are commonly used in dairy-based drinks to improve the functional properties. The World Health Organization/Food and Agriculture Organization defined probiotics as “live microorganisms which when administered in adequate amounts confer a health benefit on the host” (FAO/WHO, 2001). There is a rapid increase of using probiotic bacteria not only in beverage products, but also in different other food products due to the better understanding of the role of these bacteria in maintaining the health of the host. In addition, research and development of probiotic dairy beverages are also in progress due to the increasing demand and awareness of those products (Kandylis et al., 2016; Ranadheera et al., 2014a, 2017a; Balthazar et al., 2017). The concept of probiotic is not considered as a new notation since products produced using probiotics have been consumed for many years. At early stages of research in the areas of probiotics, there were several reports about the therapeutic properties of probiotic dairy

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Fig. 6.1  Classification of nonalcoholic beverages. (Based on Chavan, R., Shraddha, R., Kumar, A., Nalawade, T. 2015. Whey based beverage: its functionality, formulations, health benefits and applications. J. Food Process. Technol. 6 (10), 1–8.)

­ roducts where they had been using probiotic dairy products to treat p some digestive problems (Ranadheera et al., 2010). Although ­several theories on the mechanism which probiotic organisms use to exert health benefits have been proposed, the exact pathways are not fully understood yet. However, Butel (2014) suggested three modes of actions of probiotics; the first mechanism is by modulation of the host’s microbiota reducing colonization of pathogenic bacteria. Such inhibition of pathogenic may be due to the release of bacteriocins, short-chain peptides, and decreasing the pH leading for an adverse condition for bacterial growth. The second mode of action is due to the modification and improvement of the barrier properties of the gut mucosa. The third mode of action may be due to the modulation of the immune system. In addition, probiotics can synthesize enzymes such as β-­galactosidase and other enzymes which may improve digestibility of some food products. Various health benefits of probiotic consumption have been reported including alleviation of lactose intolerance symptoms, anticarcinogenic and antimutagenic activities, prevention of bacterial vaginosis and urinary tract infection, reduction in cardiovascular disease risk factors (lowering of serum cholesterol and blood pressure),

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prevention and decreasing incidence and duration of diarrhea, and maintenance of mucosal integrity (Ranadheera et  al., 2010; Franz et al., 2014). Bacteria of genera Lactobacillus and Bifidobacterium are primarily considered as probiotics. However, there are some other bacteria and yeast species which have been proved to have probiotic properties. These bacterial genera include Enterococcus, Streptococcus, and Leuconostoc (Fig. 6.2). Most of these microorganisms have been identified from different fermented milk products such as kefir, Maasai milk, and Kurut. Some probiotic lactic acid bacterium such as Lactobacillus fermentum CECT5716 could be isolated from breast milk. Most of Bifidobacterium and Lactobacillus have been isolated from the intestinal tract of health humans. In addition, some probiotic species have been isolated from the stools of healthy adults and children, body parts of animals and insects (bees, fish, and shrimps), and fermented and nonfermented food products such as meat, sausage, and vegetables (Butel, 2014). Probiotics have been used in the production of both fermented and nonfermented beverage products. Although, in many applications, probiotics have been used in combination with other starter culture microorganisms, there is a possibility to use probiotics as the only organism in the production of fermented dairy beverages (Ranadheera et al., 2016; Ranadheera et al., 2017a). Selection of a suitable probiotic strain and food matrices is very important in fermented beverage manufacturing process since it determines the efficiency of fermentation, protection of probiotics during processing, storage and gastrointestinal delivery, organoleptic properties, and the consumer acceptability. In addition, survival of probiotics in a food matrix d ­ epends on pH, ­oxygen levels, storage temperature, and the presence of competing microorganisms and inhibitors (Mattila-Sandholm et  al., 2002). Hence, it is important that manufactures pay close attention to these factors when producing probiotic beverages. Furthermore, probiotics and starter organisms used in fermented milk products may have some interactions and the strength of such interaction between these organisms depends on; whether probiotics are added during fermentation or after; the physiological state of added probiotics; and maintenance of cold chain during processing. The selected food matrices should support in maintaining the activity and viability of probiotics during storage period (Heller, 2001) and products that cannot maintain such characteristics may not be highly competitive in the market. Poor growth of many probiotics in milk limits their application in fermented milk products as some probiotic strains may not grow in milk at all (Prasanna et al., 2014). This is due to certain nutrient deficiency in milk such as amino acids for proper growth of probiotics leading to poor survival of probiotics in fermented dairy products at the ­consumption.

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Fig. 6.2  Main probiotic microorganisms. (Based on Ranadheera, C.S., Prasanna, P.H.P., Vidanarachchi, J.K. 2014b. Fruit juices as probiotic carriers. In: Elder, K.E. (Ed.), Fruit Juices: Types, Nutritional Composition and Health Benefits. Hauppauge, New York: Nova Science Publishers, 253–268; Lew, L.C., Liong, M.T. 2013. Bioactives from probiotics for dermal health: functions and benefits. J. Appl. Microbiol. 114 (5), 1241–1253; Prado, F.C., Parada, J.L., Pandey, A., Soccol, C.R. 2008. Trends in nondairy probiotic beverages. Food Res. Int. 41 (2), 111–123.)

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The recommended number of probiotics in food products including beverages at consumption should be 106–107 cfu/mL or g (Sahadeva et al., 2011). Such high viable numbers of probiotics in a food product is crucial to deliver desired health benefits for consumers. However, the shortcoming associated with the poor growth of probiotics in milk has been addressed by supplementing the milk with various ingredients. These ingredients include various sugars (glucose and galactose), protein sources (yeast extract, liver extract, peptones, and corn steep liquor), and different vi­ tamins (Sodini et al., 2005). Nevertheless, poor organoleptic properties of some of the ingredients such as yeast extract and liver extract limit their applications in fermented dairy beverage products. Therefore, there is a trend to use milk-derived ingredients such as whey protein concentrate, whey protein isolate, and casein hydrolysate to stimulate the growth of probiotics in fermented dairy beverage products (Prasanna et al., 2012; Zhang et al., 2015). These dairy-based derived ingredients can provide further development of the probiotic containing dairy beverages and strongly influences the supply of the current market where presently, there are number of dairy-based beverages already available.

6.2.1.1  Acidophilus Milk The Acidophilus milk is a fermented liquid form milk product produced by using Lactobacillus acidophilus as the starter culture. In the production process, milk is heated at 95°C for 1 h or at 125°C for 15 min. Then, cooled (45°C) milk is inoculated with 2%–5% of the pure culture of Lb. acidophilus and incubated at 37°C for 12–24 h until pH 5.5–6.0 is obtained. After the fermentation process, the product is rapidly cooled and filled into bottles, cartons, or the packaging material (Chandan, 2013; Anjum et  al., 2014). There are differences in production processes of acidophilus milk by different manufacturers. Some producers heat treat milk above 128°C which is followed by the cooling, inoculation, and the incubation. The higher temperature used in this technique helps to eliminate competitive organisms for Lb. acidophilus and support growth of the starter bacterium. In the production of unfermented acidophilus milk, cold milk (5–7°C) is inoculated with Lb. acidophilus and stored at cold conditions (Shiby and Mishra, 2013; Yerlikaya, 2014).

6.2.1.2  Bifidus Milk The Bifidus milk was first produced by Mayer in 1948 in Germany as an infant product. This milk product is considered as to be easy to digest and has been used to treat conditions such as liver diseases, gastrointestinal diseases, and constipation. During Bifidus milk processing, milk is heated at 80–120°C between 5 and 30 min and then cooled down to 45°C. The heat-treated and cooled milk is inoculated with Bifidobacterium bifidum and Bifidobacterium longum at the rate of 10% followed by incubation

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at 37°C until pH value reaches 4.5 (Yerlikaya, 2014; Kandylis et al., 2016). Once this process is completed the milk is bottled, stored, and transported at cold temperatures (around 4°C).

6.2.1.3  Acidophilus-Bifidus Milk The Acidophilus-bifidus milk is also a fermented milk, which is produced by adding concentrated probiotic bacteria to intensively heat-treated milk (Heller, 2001). In the manufacturing process, the milk is supplemented with proteins prior to fat standardization and homogenization. The standardized milk is heat treated at 75°C for 15 s (with a plate heat exchanger) or 85°C for 30 min (with the vat system) (Ozer and Kirmaci, 2009). After cooling the milk to 37°C, cultures of Lb. acidophilus and B. bifidum are inoculated and the fermentation is carried out until a pH 4.5–4.6 is reached. The fermented milk is then rapidly cooled to less than10°C and packed in appropriate packaging (Heller, 2001).

6.2.1.4 Kefir Kefir has been described as an ancient milk beverage originated many centuries ago in the Caucasian mountains (Garrote et al., 1998). In the production process, a starter culture combination containing lactic acid bacteria, acetic acid bacteria, and several genera of yeasts is used to ferment milk. These microbes are in a matrix commonly referred to as “kefir grains.” Kefir has been produced using various types of milk including sources from cow, goat, sheep, camel, and buffalo. The kefir grains have been characterized to contain bacteria such as Lac. lactis subsp. cremoris, Lac. lactis subsp. lactis, Lb. kefir, Lb. kefiranofaciens, Lb. brevis, Lb. acidophilus, Leuconostoc spp., and Acetobacter spp. Kefir grains have both lactose fermenting yeast (Kluyveromyces lactis, Kluyveromyces marxianus, and Torula kefir) and nonlactose fermenting yeasts (Saccharomyces cerevisiae) (Beshkova et  al., 2002; Tamime and Thomas, 2017). In the production of kefir, both traditional and commercial techniques are used. In the traditional method, milk is boiled, cooled to 20–25°C and inoculated with 5% of kefir grains and fermented between 18 and 24 h. After the fermentation, the kefir grains are removed by filtering with a sieve and final kefir is stored at 4°C until consumption. In the commercial manufacturing system, fat content is standardized to around 1.5%, however, the fat level may vary with the country of origin. Then, milk is warmed to 65°C and homogenized at 15 MPa. The homogenized milk is heated for 95°C for 5 min. In the production of settype kefir, the cooled milk is inoculated with a bulk starter culture at a rate between 3% and 4% (v/v) and packed in glass bottles or semirigid containers. The inoculated and packaged milk is fermented for 10–12 h

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at 19–22°C and then cooled to 9°C where the product is further ripened between 1 and 3 days. The final product is then cooled to 6°C and stored at this temperature during distribution and retailing (Wszolek et al., 2006). However, in the production of stirred kefir, the fermentate is cooled to a higher temperature (~25°C) and packaged. This step is followed by cooling to less than10°C and ripening for 15–20 h. The ripened product is cooled to 6°C and stored at this temperature throughout the distribution chain (Irigoyen et al., 2003; Wszolek et al., 2006).

6.2.1.5 Koumiss/Kumiss The koumiss is a traditional dairy drink used by nomadic cattle ­ reeders in Central Asia which contains alcohol (2%), lactic acid (0.5%– b 1.5%), milk sugar (2%–4%), and fat (2%) (Danova et al., 2005). In general, it is made from mare’s or camel’s which is inoculated by a small volume of already fermented milk. There are three types of koumiss depending on the lactic acid concentration namely strong, moderate, and light. Strong koumiss contains Lb. bulgaricus and Lb. rhamnosus and it has the pH 3.3–3.6. Moderate koumiss is fermented using Lb. acidophilus, Lb. plantarum, Lb. casei, and Lb. fermentum, and the pH of the final product is 3.9–4.5. Light koumiss is considered to have the best taste and it contains S. thermophilus and S. cremoris. In addition, some species of yeast such as nonlactose-fermenting yeast Saccharomyces cartilaginosus and noncarbohydrate-fermenting yeasts (Mycoderma spp.) responsible for the production of alcohol in the product are usually used. In the production process, the fresh milk is added to already fermented milk in a wooden container. After adding fresh milk, the mixture is stirred for around 1 h using a special wooden paddle which introduces air into the mixture (Danova et al., 2005; Wszolek et al., 2006).

6.2.1.6  Yoghurt-Based Drinks Yoghurt-based drinks are basically produced by stirring of the fermentate which effectively reduces the viscosity of the product. Yoghurt drinks are produced to fulfill the requirements of target populations. Different yoghurt-based drinks with various sugar and fat contents have been produced to fit the requirement of babies, children, and adults (Gonzalez et al., 2011). There are three main types of yoghurt drinks as outlined in the following sections. Drinking Yoghurt Yoghurt in the fluid state is commonly referred to as drinking yoghurt. In the manufacturing process of drinking yoghurt, fermentate is disturbed to produce a liquid product (Kandylis et al., 2016) and these types of products are very popular in western countries. In general, low-fat milk base, milk solids, different flavoring agents, and colors are

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used in the manufacturing process. Live starter cultures are used to produce the yoghurt coagulum and after the fermentation process the coagulum is agitated, or the cold yoghurt is homogenized to produce the final product (Tamime and Robinson, 1999). Ayran Ayran is a traditional Turkish yoghurt drink produced with adding salt and without any fruit flavoring. In the production process, the milk is standardized and fermented with S. thermophilus and Lb. delbrueckii subsp. bulgaricus (Altay et al., 2013). Sometimes exopolysaccharide producing starter cultures are used to obtain a viscous coagulum (Koksoy and Kilic, 2004). The fermentation is continued until pH 4.4–4.6 is obtained (Köksoy and Kılıç, 2003). After the fermentation, the coagulum is mixed with around 35% water and 1% (w/w) salt. Then, the mixture is churned to remove butter granules, packaged, and stored at 5°C (Tamime and Robinson, 1999). Carbonated Drinking Yoghurt The production process of carbonated drinking yoghurt is more or less similar to that of normal drinking yoghurt where CO2 is pressurized into the drinking yoghurt mixture before the packaging (Tamime and Robinson, 1999).

6.2.1.7  Cultured Buttermilk Cultured buttermilk is traditionally prepared by churning of cultured cream or cultured milk curd. However, in the modern production process, low-fat milk is fermented with the strain of thermophilic and mesophilic homofermentative bacterial cultures. It is followed by the churning of the curd and white butter is separated while the remaining liquid is called as cultured buttermilk (Mudgil et al., 2016). The culture used for the fermentation includes lactococci (Lac. lactis subsp. cremoris, Lac. lactis subsp. lactis, and Lac. lactis subsp. lactis biovar. diacetylactis) and ­aroma-producing bacteria leuconostocs (Leuconostoc mesenteroides subsp. cremoris). Furthermore, different probiotic bacteria have been successfully used in the production of different probiotic buttermilk. These include Lb. rhamnosus GG, Lb. rhamnosus 271, Lb. reuteri, Lb. casei 431, Lb. acidophilus, and Bifidobacterium spp. (Baek and Lee, 2009).

6.2.1.8  Whey-Based Drinks Whey is the main by-product of the cheese industry. Whey comprises about 85%–90% of the milk volume used in the production of cheese, and whey proteins are considered to have high biological value (Pescuma et  al., 2010). In general, whey is used in its powder form in many food preparations which is a costly application. Thus,

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ap ­ roduction of whey-based beverages can be helpful to use whey in its liquid form. Whey-based beverages can be categorized as: dairybased whey beverages, thirst-quenching beverages, and fruit juicetype beverages (Chavan et al., 2015). Dairy-Based Whey Beverages Dairy-based whey beverages are produced by fermenting liquid whey protein concentrate or milk enriched with dry whey protein concentrate and whey protein isolate (Peter et  al., 1996). Sweet whey is more suitable for the production of beverages than more acidic cheese whey, since acid whey can lead to protein sedimentation during processing. Therefore, in the production of fermented whey-based drinks, nonthermal processing techniques are used to avoid sedimentation problem (Ozer and Kirmaci, 2009). Some probiotic bacterial strains have been shown to be effective in improving physicochemical and sensory properties of whey-based drinks. The bacterial species such as, Lb. acidophilus LA-5, Lb. casei LBC-81, B. bifidum Bb-12, Lb. casei Lc-01, Lb. rhamnosus, and B. animalis subsp. lactis are common probiotic bacteria which have been used in the production of whey-based probiotic beverages (Pescuma et al., 2010; Lollo et al., 2013). Thirst-Quenching Beverages These beverages are manufactured using whey permeates which are free from proteins. One of the commercial product examples of these thirst quenching whey-based beverages includes a Swiss origin drink called Rivella (Rivella AG, Switzerland). Importantly, thirst-quenching drinks are not considered as functional food, predominately due to the thirst being potentially quenched by all other beverages and their market share is relatively small (Ozer and Kirmaci, 2009). Fruit Juice-Type Whey Beverages Acid whey is mixed with different fruit juices for the production of fruit-based beverages. Interestingly, this acid whey is a by-product of the cottage cheese manufacturing process. The acidic flavor of acid whey is compatible with acidic fruits. In addition, fruit juice-type whey beverages are fortified with vitamins and minerals (Ozer and Kirmaci, 2009; Chavan et al., 2015).

6.2.2  Soft Drinks Soft drinks represent the variety of products made using different ingredients. They can be classified based on sugar content, juice content, flavoring agent, level of carbonation, and the functionality. Table 6.1 shows the main types of soft drinks available in the market and their characteristics.

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Table 6.1  Major Types of Soft Drinks Type

Description

Bottled water

There are three subcategories. (a) Still water: noncarbonated, mineral, spring or table water, with or without added flavorings and vitamins/minerals. (b) Carbonated water: mineral, spring or table water, low carbonated waters, naturally sparkling or sparkling by CO2. (c) Flavored water: unsweetened water, with essences and/or aromatic substances. Potable water is sold in packs of over 10 L for use in dispensers. Sweetened, beverages with CO2, syrups for home dilution and out-of-home carbonated soft drinks. 100% pure fruit or vegetable juice without ingredients, except permitted minerals and vitamins with sweetening agents (less than 2%). Diluted fruit/vegetable juice and pulp, with sweetening agents, minerals, and vitamins. Flavored ready-to-drink, noncarbonated beverages, containing fruit or nonfruit flavors or juice content (up to 25%). Nonready-to-drink products, marketed as concentrates for home consumption including fruit and nonfruit-based products and flavors. Nonready-to-drink products in powder form. Tea-based or coffee-based drinks and nonready-to-drink powders and liquid concentrates for dilution. Products described as “isotonic”, “hypertonic”, or “hypotonic”, still or carbonated, readyto-drink, or nonready-to-drink powders and concentrates; also fruit and nonfruit flavoued drinks. Energy-enhancing drinks, mainly carbonated and containing taurine, guarana, glucose, caffeine, exotic herbs, and substances, minerals and vitamins

Bulk/hot water Carbonated drinks Juice Nectars Still drinks Squash/syrups Fruit powders Iced/ready-to-drink tea/ coffee drinks Sports drinks

Energy drinks

(From Kregiel, D. 2015. Health safety of soft drinks: contents, containers, and microorganisms. Biomed. Res. Int. 2015, 15. https://doi. org/10.1155/2015/128697.)

6.2.2.1  Carbonated Drinks Carbonated beverages are drinks that contain dissolved CO2 for var­ ious reasons. From the consumer perspective, many find a fizzy sensation to be pleasant and like the slightly different taste that carbon dioxide provides. Carbonation process produces the characteristics fizziness and bubbling in these drinks and this is due to the dissolved CO2 in a liquid under pressure. However, the carbon dioxide used must be free of odor and flavor (Johnson et al., 2010). The main ingredients used in carbonated drinks are water, carbon dioxide, sweeteners, flavoring, colors, and acids. The sweeteners may be nutritive sweeteners such as sucrose and fructose or low calorie nonnutritive sweeteners (Potter and Hotchkiss, 2012). Carbonated drinks can be classified into d ­ ifferent groups based on

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many aspects. There are nonflavored carbonated beverages, flavored carbonated beverages with natural extracts, flavored carbonated beverages with artificial flavors, and carbonated beverages with fruit juice (Saint-Eve et  al., 2010). In the production of a carbonated beverage, the concentrated flavoring (beverage base) is combined with a nutritive or nonnutritive sweetener and water to form a syrup. The syrup is mixed with a proportioned quantity of carbonated water followed by filling and sealing the beverage in a container (NPCC, 2012).

6.2.2.2  Fruit Juice Fruit juice is an unfermented but fermentable liquid commonly obtained from an edible part of the fruit. The juice is prepared by a suitable process which maintains the essential physical, chemical, organoleptic, and nutritional characteristics of the juices of the fruit from which it comes. In the production of fruit juices, fresh frozen or chilled fruits are used as the raw material depending on the availability and desired quality parameters of the final product. Main steps followed in the juice production include extraction of juice, deaeration, pasteurization, concentration, adding back essence, and packing (Potter and Hotchkiss, 2012; Ashurst, 2013).

6.2.2.3  Hot Drinks Tea, coffee, and chocolate-based beverages are common hot drinks which are served at temperatures between 71.8°C and 85.8°C (Brown and Diller, 2008). Tea Tea is made from leaves of Camellia sinensis. Tea has hundreds of bioactive substances such as amino acids, caffeine, lignins, proteins, xanthines, and flavonoids (Yadav and Mendhulkar, 2015). The flavonoids of tea are responsible for health benefits (Vinson et al., 1995). Tea can be classified based on the processing technique used during manufacturing, and six types have been identified on that principle, namely green tea (nonfermented), yellow tea, white tea, oolong tea (semifermented), black tea (fully fermented), and dark tea (Chen et al., 2009). The manufacture of black tea involves the oxidative polymerization of the monomeric flavan-3-ols by the enzyme polyphenol oxidase. Therefore, to produce black tea, plucked leaves are rolled and fermented for 90–120 min (Rusak et al., 2008). In contrast, during the production of green tea, freshly harvested tea leaves are quickly heat treated (steamed or hot air dried) to inhibit the oxidizing enzyme and therefore, prevent the fermentation and maintain its characteristics green color (Zhao et al., 2006; Naumovski, 2015; Naumovski et al., 2015). Yellow tea is produced similarly to green tea; however, the production

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process has a slower drying period which allows the damp tea leaves to become yellow (Wang et al., 2013). White tea is manufactured using very young tea leaves or buds covered with tiny slivery hairs. In the processing, plucked tea leaves are immediately steamed and dried to prevent oxidation resulting in a light delicate taste (Rusak et al., 2008). Coffee The coffee tree comes under the family Rubiaceae and there are more than 70 species have been identified. Coffea arabica (Arabica) and Coffea canephora (Robusta) are the main commercially cultivated species, while the former accounts for around 75% of world production (Belitz et al., 2009). Usually, the dry or wet method is used to convert coffee cherries into green coffee beans. In the dry method, coffee cherries are dried and the dried pericarp is mechanically removed to produce green coffee or natural coffee. However, the wet method is used to produce parchment coffee where the exocarp of cherries is initially removed mechanically in the presence of water. The mesocarp is removed by fermentation or other methods followed by washing and drying resulting parchment coffee (Batista et al., 2009). In the retail market, many forms of coffee are available such as of green coffee bean, whole roasted coffee bean, ground coffee, and brewed coffee drinks (Bhumiratana et al., 2011). Cocoa-Based Beverages Powdered cocoa-based beverages are produced by mixing cocoa powder with sugar and/or milk powder (Shittu and Lawal, 2007). In general, a cocoa powdered beverage is made of 80% sugar and 20% of cocoa. Some producers use different ingredients such as fat, water, soluble micronutrients, emulsifiers, and stabilizers to improve functionalities of their products (Abdelaziz et al., 2014). Furthermore, the addition of ingredients other than coca is vital to quality of the final product. Consequently, the selection and addition of these ingredients should be fully considered as some of these ingredients will pose challenge to the manufacturing processes (Mellor et al., 2017).

6.3  Nutritional Value of Nonalcoholic Beverages 6.3.1  Nutritional Aspects of Consumption of Dairy-Based Beverages Several factors determine the nutritional value of dairy-based beverages. These factors include the type of milk, supplementation, the type of microorganisms, and the manufacturing process and more specifically whether the product is fermented or not (Shortt and O’Brien, 2004).

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Consumption of milk-based beverages can supply many nutrients required for the human. Milk is a rich source of protein with an average content of 32 g/L. Caseins represent around 80% of milk proteins, while soluble whey proteins represent the leftover of the milk proteins (around 20%). These proteins are considered to be high-quality proteins having high digestibility and bioavailability (Séverin and Wenshui, 2005) and containing all essential amino acids in their structure. Bioactive peptides resulting from hydrolysis of milk proteins have been shown to have various biological roles leading to health benefits. These peptides have been reported to be effective as antifungal, antiviral, antioxidant, antihypertensive, and immunomodulatory agents (Boye et  al., 2012; Pereira, 2014). In addition to macronutrient profile, milk is considered as a good source of minerals predominately calcium, magnesium, potassium, zinc, and iron (Popkin et al., 2006). Milk is also a good source of some vitamins such as A, D, and E and thiamine, riboflavin, and vitamin B12. Sometimes, milk is fortified with vitamin A and D to improve its nutritional properties particularly after processing (Gaucheron, 2011). Milk fat can play a vital role as an energy source in human nutrition. In addition, milk fat is a good “carrier” of some vitamins such as vitamin A, D, and E. Milk is considered as a source for essential fatty acids namely linoleic and linolenic acid although cows’ milk contains a very low amount (~0.3%–0.6% of total fatty acids). These essential fatty acids can serve as precursors for some hormones and other essential metabolites (Dhiman et al., 2000; Walstra et al., 2005). Milk fats are a complex mixture of lipids where the most of fats exist in the form of triglyceride. Fatty acids present in milk are primarily saturated and monounsaturated. The level of polyunsaturated fatty acids in milk is very low. There is a consumer concern about saturated fat intake from milk since high intakes of saturated fat can potentially lead to increased total plasma cholesterol and low-density lipoprotein (LDL) cholesterol leading to elevated risk of cardiovascular disease (Gibson, 2011). In general, fermented milk products are characterized by a lower level of residual lactose, and higher levels of free amino acids and certain vitamins than nonfermented milk and therefore, fermented milk products are suitable for people who have lactose intolerance (Gomes and Malcata, 1999). In addition, some probiotic bacteria such as bifidobacteria have been reported to be capable of synthesizing and liberating many kinds of vitamins, such as thiamine, nicotinic acid, pyridoxine, vitamin B12, and vitamin K (Tamime et al., 1995; Hou et al., 2000). In addition, it was reported that the probiotic bacterial strain L. fermentum D3 could increase the in vitro bioavailability of Ca, P, and Zn in fermented goat milk (Bergillos-Meca et al., 2013).

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6.3.2 Importance of Soft Drinks in Human Nutrition Water is very important in human nutrition not only from the hydration and “thirst quenching” perspective, but it is also considered as the major constituent of the human body (Jéquier and Constant, 2010). Water is essential for metabolic activities and physiological functions and it is considered as the primary fluid in the human body which serves as a solvent for minerals, vitamins, amino acids, glucose, and other nutrients. In addition, it has a vital role in the digestion, energy production, joint lubrication, absorption, transportation, and use of nutrients. Furthermore, water is used as the medium for the elimination of toxins and waste products and thermoregulation of the body (Kleiner, 1999; Popkin et al., 2006). Drinking water can have an effect on the weight of consumer by reducing caloric intake since drinking water can help people to feel fuller at mealtimes (Burls et al., 2016). Soft drink consumption is becoming a controversial public health and policy issue in almost all the countries around the world. Sweetened energy dense beverages include carbonated fizzy drinks and still beverages, which are usually sweetened with fructose or sucrose (Popkin et al., 2006). The consumption of soft drinks is considered to be one of the major reason for obesity and health-related problems among adults and children (Vartanian et al., 2007). Sweetened caloric beverage consumption was shown to increase energy intake, body weight, fat mass, and blood pressure in adults (Raben et al., 2002). However, noncaloric beverages are preferred over sweetened caloric beverages by some consumers since they provide water and sweetness without the added energy intake (Popkin et al., 2006). Consumption of fruits, vegetables, and their juices (in the unsweetened form) can have a favorable effect against inflammation, coronary artery disease, and incidence of various cancers primarily due to vitamins and phytochemicals such as phenolic compounds and flavonoids (Aptekmann and Cesar, 2013; Perche et  al., 2014). It had been considering that 100% fruit juices are a healthy alternative to sugar-sweetened beverages since they have some important nutrients. Nevertheless, fruit juices have the higher amount of natural sugars resulting in high number of calories to the consumers and therefore, they need to be consumed at a moderate level. A positive relation between regular consumption of fruit juices and weight gain of consumers was observed and therefore, it is recommended to limit 100% fruit juice consumption to no more than 120–180 mL a day (Bazzano et al., 2008; Hu, 2013). However, vegetable juices can be healthy alternatives to fruit juices since they contain fewer kilojoules per unit volume than that of fruit juices predominately due to the lower sugar content (Popkin et al., 2006).

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6.3.3 Effect of Consumption of Hot Beverages on Human Health Tea is a beverage which provides antioxidants, flavonoids, some micronutrients, and some soluble fiber, but no calories (du Toit et al., 2001; Popkin et al., 2006). Polyphenols in tea can react with proteins via hydrophobic interactions and hydrogen bonding leading a formation of sediment. In addition, polyphenols can change molecular configuration of enzymes leading to loss of catalytic activity (He et al., 2007). Metal ions are easily bound with tea polyphenols which may lead to poor absorption of metal ions causing some health problems such as infant microcyte anemia (Yang and Landau, 2000). However, these findings are predominately in populations of individuals with marginal iron status. On the contrary, it is known that regular consumption of tea (in particular green tea) can be associated with reduced risk of cardiovascular diseases and some forms of cancer (Yang and Landau, 2000; Popkin et al., 2006). Coffee is considered as a beverage which can provide many bioactive compounds. Coffee contains carbohydrates, lipids, nitrogenous compounds, vitamins, minerals, alkaloids, and phenolic compounds. Some of the nutrients found in coffee such as magnesium, potassium, niacin, and vitamin E could potentially lead to observed health effects with coffee consumption (Higdon and Frei, 2006). A caffeine, natural alkaloid in coffee beans, is very important chemical related with many biological functions including stimulation of central nervous system, acute elevation of blood pressure, the increase of metabolic rate, and diuresis (Ranheim and Halvorsen, 2005; Higdon and Frei, 2006). In some studies, it has been shown that high intake of coffee is associated with the reduction of colorectal cancers (Popkin et  al., 2006). However, coffee consumption can negatively affect human health via the generation of higher levels of amino acid homocysteine. An elevated homocysteine concentration in plasma is considered to be an independent risk factor for cardiovascular disease and a positive correlation between consumption of coffee and plasma homocysteine concentration was reported by Ranheim and Halvorsen (2005). Ground, roasted, shelled, and fermented cocoa beans are a paste commonly referred to as “cocoa liquor” and it has both nonfat cocoa solids and cocoa butter. Cocoa butter has a significant amount of fatty acids, while the nonfat cocoa solid contains many vitamins, minerals, polyphenols, and fiber (Katz et  al., 2011). The polyphenols in cocoa include catechins, procyanidins, and anthocyanidins which can exhibit antioxidant properties in vitro (Richelle et al., 2001). Cocoa consumption has been shown to improve lipid profile, improve insulin sensitivity, reduce platelet activity and function, ameliorate endothelial dysfunction, and diminish blood pressure (Monagas et al., 2009).

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A number of potential mechanisms have been proposed on the effect of cocoa polyphenols in improvement of health in humans. One of the mechanisms to potentially explain the effect of cocoa flavanols in the reduction of blood pressure is via its effect on nitric oxide. This potential mechanism proposes that the flavanols induce endothelial nitric oxide synthase resulting in the increase in the available pool of nitric oxide (Mellor et al., 2017).

6.4  Sources and Occurrence of Pathogenic Microorganisms in Nonalcoholic Beverage Products Despite technological innovations, nonalcoholic beverages products may be subject to microbiological contamination when the appropriate hygiene and sanitation conditions are not met. The hygiene of the equipment and utensils used in the processing and the hygienicsanitary conditions of the manipulators are fundamental to obtain products with microbiological quality. Table 6.2 shows the main nonalcoholic beverages, the processing steps where there may be microbiological contamination and the main microorganisms involved. Fruit juices are unfermented liquids obtained from the edible part of healthy and ripe fruits, while the concentrated juices are juices from which the water has been physically removed. Fruit nectars are unfermented pulp-based beverages, prepared with one or more fruits, to which sweeteners and other ingredients may be added (ICMSF, 2015). Most fruit-based products undergo thermal pasteurization or commercial sterilization treatments and have low pH; therefore, the microorganisms present are hardly harmful to human health. However, in the last decades, some fruit juices have been linked to outbreaks caused by Escherichia coli, Salmonella sp., and Cryptosporidium. These fruit juice-related outbreaks changed the belief that acidic foods do not require concern regarding the delivery of pathogenic microorganisms (Vantarakis et al., 2011). Microbial contamination of fruit drinks is usually due to the exposure of fresh fruits or industrialized products to microorganisms present in the harvesting, storage, and transportation environments and where there are failures in thermal processing and/or contamination after processing. Unpasteurized juices have attracted the interest of the consumers because of the greater freshness and the maintenance of the color, flavor, vitamins, and minerals present in the raw fruit. This type of product is more susceptible to microbiological contamination and may be a vehicle of pathogens to humans. A study by ­Jackson-Davis et  al. (2018) evaluated the outbreaks related to

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Table 6.2  Main Ingredients and Processing Steps Involved in Microbiological Contamination of Nonalcoholic Beverages Type of Beverage

Processing Step

Associated Microorganism

Fruit or vegetable drinks

Raw material (type of fruit) Water Harvest Inefficient heat treatment Ingredients (water, sugar, syrup, fruit juice, flavorings, preservatives, acidulants) Post cooling Packing Raw material (leaves) Ingredients (water, fruit juices, sweeteners, milk, soy protein) Inefficient heat treatment Raw material (milk) Ingredients (cocoa powder, sugar, fruit concentrates, thickeners, flavorings) Inefficient heat treatment

Salmonella sp., E. coli 0157:H7, Clostridium botulinum, Trypanosoma cruzi

Carbonated or noncarbonated soft drinks

Tea-based drinks

Dairy drinks

Salmonella, E. coli, Chryseobacterium meningosepticum, Klebsiella spp, Staphylococcus, Stenotrophomonas, Candida, and Serratia Clostridium botulinum, Salmonella

Listeria monocytogenes, Staphylococcus aureus, E. coli, Salmonella spp.

fruit juice ­consumption ­between 1922 and 2014 and reported that most of the cases were related to the consumption of unpasteurized fruit juices. The major reported pathogenic microorganisms were: Trypanosoma cruzi, Salmonella Typhi, Salmonella typhimurium, E. coli O157: H7, Cryptosporidium parvum, and Clostridium botulinum. Similarly, a study by Vojadani et  al. (2008) compiled the outbreaks of illness associated with the consumption of fruit juice from 1995 to 2005, and recorded 21 juice-associated outbreaks reported to the CDC. These outbreaks included, 10 cases related to apple juice or cider, eight linked to orange juice, and three involved other types of fruit juice. These outbreaks caused 1366 illnesses, with a median of 21 cases per outbreak (range, 2–398 cases). Among the 13 outbreaks of known etiology, five were caused by Salmonella, five by E. coli O157: H7, two by Cryptosporidium, and one by Shiga toxin-producing E. coli O111 and Cryptosporidium. Soft drinks are carbonated or noncarbonated products, contain rel­ atively typical list of ingredients (water, sugar, flavorings, p ­ reservatives,

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and acidulants), which can be added to fruit juices, fruit pulps, or fruit peels. Noncarbonated beverages are predominantly fruit-based, noncarbon dioxide added and are generally heat treated or added with chemical preservatives (ICMSF, 2015). Soft drinks are a medium not suitable for microbial growth because they have a high content of carbon dioxide (1.5–4 volumes), high acidity (addition of acidulants such as citric or phosphoric acid), low pH (2.5–4), and contain preservatives (potassium sorbate or potassium benzoate). However, the presence of sugars may represent an important source of energy for these microorganisms (Azeredo et al., 2016). The first precaution with the microbial safety of soft drinks should be the water quality. The bacteria found in water are generally of the genera Pseudomononas, Chromobacterium, Proteus, Achromobacter, Microccoccus, Bacillus, Streptococcus, and Aerobacter. However, pathogenic bacteria like Salmonella and E. coli can also be carried by water. Therefore, water to be used in soft drink processing must be pretreated to ensure its safety. Water treatment usually involves chlorination, softening, flocculation and subsequent separation of particles (by flotation or sedimentation), slow sand filtration, super chlorination, activated carbon filtration, and polishing filtration (Azeredo et al., 2016). Other raw materials, such as sugar, may be contaminated with bacteria, molds and yeasts, as well as spores, due to the growth of thermophiles in some steps during processing. Similarly, simple syrup may represent a microbiological hazard to soft drinks, since some thermosetting spores and/or molds may survive the temperature ranges used during heat treatment (80–85°C). The main critical point in the soft drink processing line is in the step after cooling, mainly in the piping between the single syrup cooler and the compound syrup preparation tank. The recommendation is not to store this syrup for long time and some industries add preservatives and acidulants for better conservation. Care should also be taken regarding the microbiological quality of the fruit juices that will be added to the products and of the other ingredients, such as flavorings, dyes, acidulants, and among others. Finally, the filling stage and packaging are important, requiring clean and sanitized equipment and packaging materials. The packaging process and used materials should be free from adhesion of microorganisms (biofilms). White et al. (2010) reported that opportunistic pathogenic microorganisms, such as Chryseobacterium meningosepticum, Klebsiella sp., Staphylococcus, Stenotrophomonas, Candida, and Serratia can be persistent contaminants of soda fountain machines (White et al., 2010). Ready-to-drink tea-based beverages include products that are produced by direct leaf extraction, sweetened and flavored, as well as carbonated soft drinks made from instant tea solids and lemon juice,

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which are low in pH and are preserved by weak acids (ICMSF, 2015). In general, the heat treatment of pasteurization significantly reduces the concern with the safety of this type of product. As mentioned for the soft drinks, the quality of the tea-based drink should be controlled, as well as that of other ingredients that may be used, such as flavoring and sweeting ingredients. Dairy drinks can be fermented or nonfermented products. The fermented products (yoghurts and fermented milks) have low pH, are made of pasteurized milk, and the starter cultures are competitive microbiota for the pathogenic microorganisms. Unfermented dairy drinks are heat-treated products, subjected to pasteurization or ultra-high temperature (UHT), with different flavors and marketed in plastic, glass, or Tetra Pak containers. Both categories (fermented and nonfermented dairy drinks) have good nutritional quality, and can be a healthy alternative as a quick snack for children and adolescents. Unfermented dairy drinks are also fortified with cheese whey. In general, the presence of pathogenic microorganisms in this type of product is related to postprocess contamination or failure in the heat treatment stage. The care that must be taken is related to the raw material (milk) and ingredients used in the processing, such as cocoa powder, sugar, fruit concentrates, thickeners, flavorings, etc. In addition, it is of utmost importance to consider spore-forming microorganisms, mainly in the UHT products. In a general view, the nonalcoholic beverages are not products with high susceptibility to pathogenic contamination, when the basic hygienic conditions of good hygiene practices and the control of the critical points on the processing in the industry are followed. Therefore, a control of the hygiene of equipment, process, environment, and packaging is primordial, and the microbiological control must consider all aspects related to the water and ingredients used in processing, and the processing steps, mainly the heat treatment. A special attention should be given to products unpasteurized, without preservatives, with low carbonation and with high fruit juice contents. Furthermore, the intrinsic characteristics of the raw material, like the pH of the fruit juice, must be considered, as some juices may not fall into the high acid dogma of the more commonly known juices. The nonalcoholic beverage products are generally considered of low pathogenic potential when processed and handled properly. The low pathogenic potential of these products can be confirmed in a search of foodborne outbreak from 1998 to 2016 considering fruit juices, dairy drinks, soft drinks, and tea beverages. To date in the United States, three outbreaks were reported, with 18 illnesses, four hospitalizations, and one death. The foods involved were pasteurized carrot juice, kale and pineapple unpasteurized juice, and yoghurt (CDC, 2017).

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6.4.1 Common Pathogenic Microorganisms That May Be Associated With Nonalcoholic Beverage Products 6.4.1.1  E. coli O157:H7 E. coli belongs to the family Enterobacteriaceae, genus Escherichia, being found in the human and mammal gut. Bacteria of this genus are mobile or immobile, gram negative, aerobic or anaerobic facultative, and nonsporulated. They present optimal growth temperature at 37°C and optimum pH close to the neutrality (pH 7). E. coli O157: H7 belongs to the enterohemorrhagic group, which causes bloody diarrhea, hemorrhagic colitis, hemolytic uremic syndrome (HUS), and thrombotic thrombocytopenic purpura. The onset of symptoms occurs after 1–2 days of consumption of the contaminated food (Ray and Bhunia, 2007; Ranadheera et al., 2017c; Yu et al., 2017). The presence of E. coli O157: H7 in unpasteurized apple juice was reported in the United States as responsible for the hospitalization of 25 people, 14 with development of HUS, and one case of death (Cody et  al., 1999). Contaminations were also reported for unpasteurized apple cider (Food Safety Chart, 2010; Canadian Food Inspection Agency, 2014). Considering the trends in fruit juices market, the exotic juices, such as acai, melon, persimmon, and papaya, which have low acidity (pH 4.8–6.2), provide suitable conditions for survival and growth of pathogenic bacteria. The increased commercialization of soft drinks without preservatives, with low carbonation and with high fruit juice volumes, resembling the natural fruit juices, are more associated with foodborne pathogens (Juvonen et al., 2011). Doogh is a traditional Iranian dairy drink, prepared by dilution of low-fat yoghurt with water and further fermentation to achieve satisfactory taste and acidity, or occasionally by bacterial fermentation of milk with yoghurt culture; with the addition of salt and flavoring. Zarei et al. (2015) monitored the behavior of E. coli O157:H7 in traditional Iranian doogh at different temperatures and time periods. E. coli O157:H7 was detected in low acid doogh sample after 10–14 days at 4°C, but just only 2 days at 25°C. In contrast, in high acid doogh samples, the viability declined more quickly to undetectable level, both at 4°C and 25°C indicating less viability of these bacteria in high acid doogh samples. The same authors concluded that this matrix should be considered as a potential vehicle of transmission of this pathogen, especially in low acid doogh samples stored under refrigeration.

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6.4.1.2 Salmonella Bacteria of the genus Salmonella are pathogenic to humans and animals and belong to the family Enterobacteriaceae. They are shaped like cocci, bacilli, or short rods of various sizes, are mobile, gram negative, facultative anaerobic, and catalase positive. Salmonellosis is a foodborne infection caused by the viable cells of Salmonella sp., resulting in nausea, vomiting, abdominal pain, and fever, with an average incubation period between 12 and 24 h with symptoms that may remain for up to 14 days. They multiply well in pH near to the neutrality and in the temperatures of 35–43°C. Salmonella is a major cause of foodborne diseases worldwide with significant morbidity, mortality, and economic losses. Salmonella is a problem mainly in fresh and unpasteurized fruit juices, due to its low thermal tolerance (Koch et al., 2005; Jain, 2006; Ray and Bhunia, 2007; Ranadheera et al., 2017b,c). In October 1974, hospitals in Bergen County, New Jersey, USA, reported the isolation of S. typhimurium from 13 patients who had gastroenteritis after drinking cider. Local publicity resulted in the identification of 296 people who had suffered similarly (CDC, 1975). In 1999, Salmonella serotype Muenchen infections were observed in consumers of a commercially distributed unpasteurized orange juice, comprehending 207 confirmed cases reported by 15 states and two Canadian provinces. The predominant symptoms reported were diarrhea (94%), fever (75%), and bloody diarrhea (43%). Many patients were hospitalized, and no patients died (CDC, 1999). In 2000, an outbreak of 88 cases of culture-confirmed Salmonella enteritidis infection was associated to orange juice. Juice from one plant was transported to another for mixing and bottling and it was identified several potential hazards (Rangel, 2000). In 2005, officials in 23 states reported 152 cases of S. typhimurium infection associated with commercially distributed unpasteurized orange juice (Jain, 2006). The main risk factors of the outbreaks were associated with the fertilization of agricultural crops: crops were grown in orchards where sheep grazed, manuring the soil. Fallen fruit were often used, allowing soil and faecal contamination. In addition, poor decontamination of fruit occurred in the factory; poor-quality wash water was used; pest control within the factory was poor; and the cleanup of fruit boxes and conveyors was poor (Wareing and Davenport, 2007). Akond et al. (2009) evaluated a total of 225 carbonated soft drink samples from nine brands, from various locations in five metropolitan cities of Bangladesh. In all, 54% of the samples yielded Salmonella spp. at numbers ranging from 2 to 90 cfu/100 mL, suggesting that carbonated soft drinks commercially available in Bangladesh pose substantial risks to public health. The results are presumably originated from contaminated raw materials or from water, and the microbial survival was due to the poor manufacturing processes.

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Contaminations of tea beverages are not common, but, tea leaves and infusions were related to outbreaks in the past years. A Salmonella Agona outbreak affecting infants (42 children below 13 years of age) in Germany (2002–2003) was caused by aniseed-containing herbal tea imported from Turkey (Koch et al., 2005). Baby tea is widely distributed throughout Serbia and contains aniseed, caraway, and fennel seeds. Sanitation inspectors collected samples from tea manufacturers and, in the fennel seed sample, S. Senftenberg was identified (Ilic et al., 2010).

6.4.1.3  Listeria monocytogenes Listeria is a Gram-positive, nonspore-forming bacterium. It is mobile by means of flagella and classified as psychrophilic, which can grow and multiply even at low temperatures (0–15°C). Listeria monocytogenes is a pathogen and presents rod-shaped cells, being microaerophilic and capable of multiplying at refrigeration temperatures. It causes the development of flu-like symptoms such as fever, headaches, vomiting, and nausea. In chronic cases, it is related to septicemia in pregnant women, fetuses or newborns, internal or external abscesses, meningitis, etc. L. monocytogenes has not yet been implicated in juice-related outbreaks, but it has been shown to be capable of longterm survival in various frozen juice concentrates (Juvonen et al., 2011). The reason why there are no reports on listerioses (a ubiquitous pathogen) linked to the consumption of fruit or fresh juices, in contrast to the variety of outbreaks related to enteropathogens, is unclear (Tribst et al., 2009). However, Sado et al. (1998) did a microbiological survey of 50 retail juices in 1996 and observed that two unpasteurized juices were positive for L. monocytogenes: an apple juice and an apple raspberry blend with a pH of 3.78 and 3.75, respectively. Sheela and Shrinithivihahshini (2017) evaluated milk and dairy products (n = 415) from Tiruchirappalli city, Tamil Nadu, India, considering the incidence of L. monocytogenes. L. monocytogenes were isolated from 219 (52.7%) samples. Among the positive samples, the raw milk and flavored milk were 100% contaminated followed by branded milk (65.9%), cheese (62.5%), ice-cream (49.2%), milk powder (26.6%), milk sweets (20%), ghee and paneer (13.3%), and yoghurt (6.6%). Conversely, curd and butter were free from L. monocytogenes. The authors concluded that the milk and dairy products were vulnerable for L. monocytogenes and it is suggested to create awareness among people, especially elderly people and children, about this pathogen.

6.4.1.4  Clostridium botulinum Bacteria of the genus Clostridium are rod shaped and move by p ­ etritic flagella, strictly anaerobic, Gram-positive, mesophilic or t­ hermophilic, and catalase positive. For their development, they require weakly acid

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or alkaline medium, temperature between 19°C and 37°C and nutrients such as amino acids, minerals, and vitamins. Consequently these bacteria can produce toxin, causing the botulism. Symptoms of botulism appear 12–36 h after ingestion of the neurotoxin and associated with nausea and vomiting followed by neurological disorders such as double vision, fixed and dilated pupils, difficulty of speaking and swallowing, dry throat and tongue, throat pain, fatigue and loss of muscle coordination, and respiratory failure, which can lead to death within a few days (Ray and Bhunia, 2007). In 2006, four cases of botulism linked to carrot juice occurred in the United States (CDC, 2006). The products involved were pasteurized, but not at temperatures that eliminated C. botulinum spores. The carrot juice products involved in these illnesses were distributed under refrigeration with one or more of the following label statements “Keep Chilled,” “Keep Refrigerated,” “Perishable Keep Refrigerated,” or “Extremely Perishable Keep Refrigerated.” Because proteolytic C. botulinum spores are known to grow and produce toxin only under severe temperature abuse conditions, the juice involved in these outbreaks may have been left unrefrigerated for an extended period, either during distribution or while being held by consumers, allowing C. botulinum spores to grow and produce toxin. One of the measures of control of C. botulinum is the maintenance of the products in temperatures of refrigeration and the acidification to pHs lower than 4.6. Considering the dairy products, the botulism outbreaks are rare compared to those linked to vegetable, meat, and fish, and extremely rare compared to disease caused by other foodborne pathogens, however, they may be large and have serious consequences. The vehicle of botulism was cheese or processed cheese in 14 dairy outbreaks (70%), milk in three (15%), yoghurt in two (10%), and dried milk powder in one (5%). At least 12 of the outbreaks were due to commercial products (Lindström et  al., 2010). The first outbreak related to yoghurts occurred in the United Kingdom in 1989. It involved 27 patients and one death. The contaminated products were hazelnut yoghurt and the hazelnut conserve (O’Mahony et al., 1990).

6.4.1.5  Staphylococcus aureus Species of the genus Staphylococcus are cocci, Gram-positive, facultative, and nonspore-forming bacteria (Ray and Bhunia, 2007). They have a nutritional need for amino acids and B vitamins, such as thiamine and nicotinic acid, their optimum growth temperatures, and pH are 7–48°C and 6–7, respectively. They are common in the nasal and oral mucosa of man and certain animals, as well as in the skin, hair, and infections (wounds and tumors). The toxic action is performed through its enterotoxin, with incubation time of 1–6 h and with symptoms such as: nausea, vomiting, abundant salivation, spasms and

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a­ bdominal pains, sweating, chills, muscle cramps, and diarrhea. Fatal cases are rare and are generally associated with elderly people and patients with cardiovascular and immunocompromised diseases. Considering fruit juices, Staphylococcus aureus is mainly found as a contaminantinstreetvendedproducts(Tambekeret al.,2009;Reddiet al.,2015). Reddi et al. (2015) evaluated a total of 150 samples of street vended fruit juices like grapes, pineapple, sapota, and sweet lime. About 96.6% of the tested fruit juices were contaminated with various microorganisms, including fecal coliforms (77.3%), S. aureus (73.3%), Shigella spp. (48.6%), and E. coli (42.6%). The findings concluded that educating and training vendors handling foods are essential in order to improve food safety. It is also essential to change behavior and communication techniques to create awareness among vendors and translate into practice to avoid possible sources of microbial hazards. Another study by De Buyser et  al. (2001) reported that S. aureus was involved in 15% of recorded foodborne illnesses caused by dairy products in eight developed countries and was responsible for more than 85% of the dairy borne diseases in France. However, yoghurt was regarded as hygienically safe against pathogens due to milk pasteurization and high acidity of yoghurt which were thought to be effective barriers against contamination by S. aureus.

6.4.1.6 Mycotoxins Several species of fungi produce biological metabolites called mycotoxins that, by contaminating food, make them toxic to the body. The growth of filamentous fungi in fresh fruits and their juices may lead to the formation of mycotoxins such as patulin and ochratoxin. Patulin is found mainly in apple and pear juice and is produced by Penicillium, Aspergillus, and Byssochlamys, while ochratoxin is found in grape juice and is produced by Aspergillus caebonarius or Aspergillus niger and related species (ICMSF, 2015). Patulin is a carcinogenic compound easily transferred from fruit to juice because it is soluble in water. Although some steps in juice processing may reduce the existing quantities, pasteurizing would have no significant effect. Therefore, rotten, windblown, or damaged fruits should not be used for the processing of fruit juices. In addition, an initial stage of water treatment and storage of the fruits under refrigeration is essential. In the case of unpasteurized fruit juices and fruits with low acidity, such as tomatoes and melons, refrigeration serves as an additional barrier to reduce the risk of development of pathogenic microorganisms. Studies in various countries revealed the presence of patulin at concentrations higher than the international recommendations (50 μL/L). For examples, the detected amounts exceeded the recommended level by 11% in the United States (Harris et al., 2009) and Spain (Murillo-Arbizu et al., 2009), 16% in China (Yuan et al., 2010), and 33.8% in Pakistan (Igbal et al., 2018).

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6.5  Risk Management and the Strategies to Minimize the Foodborne Illnesses Related to Nonalcoholic Beverages The microbial risk assessment is involved in an estimation of the magnitude of human health risk in terms of likelihood of exposure to a pathogenic microorganism in a food and the likelihood and impact of any adverse health effects after exposure. Risk assessment for evaluating and managing foodborne microbiological health risks comprises many steps including hazard identification, hazard characterization, exposure assessment, and risk characterization (Lammerding, 1997). This framework has been adopted by the Codex Alimentarius Commission, the internationally recognized standard-setting body for foods in international trade (CAC, 1999; Lammerding and Fazil, 2000). These approaches, therefore, provide an objective scientific basis for risk management and decision making in ensuring the safety of the food products. Unlike chemical or physical contaminant hazards, microbiological hazards can be introduced at any step in the food and beverage production chain and the most effective opportunity for controlling those hazards can very well be a different step. Microbial risk management thus requires a systematic and detailed understanding of the entire food production chain (Havelaar et  al., 2010). In this regard, monitoring the presence of pathogen in the final product is not adequate in risk management and additional necessary protection approaches should be employed from the beginning of the production. Usually, a proactive approach is recommended, starting with the producer ensuring a safe product and process design, and predicting where problems might arise, rather than detecting them after they have occurred (Havelaar et  al., 2010; Ranadheera et  al., 2017c). Nowadays, various systems including hazard analysis critical control point (HACCP) programs and good manufacturing practice (GMP) are widely used in managing microbiological hazards in food and beverages as these approaches have demonstrated highly effective for the control of these hazards. In addition, there are many food laws and regulations imposed by national governments and international agencies to ensure the safety of food and beverage products. The risk assessment approaches can be either qualitative (descriptive or categorical treatments of information) or quantitative (mathematical analyses of numerical data) (Lammerding and Fazil, 2000). Some of these current trends used to analyze and support decision making on food safety include microbial risk assessment models, predictive microbiology, dynamic infectious disease models, risk factor models, attribution models, and multi-criteria analysis models (Havelaar et al., 2010).

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6.6 Conclusions The global nonalcoholic beverage market represents a significant segment in the food and beverage industry at present and will ­continue to grow substantially in coming years. Nonalcoholic beverages can also be considered as one of the key contributors for foodborne illnesses due to many factors such as the diversity, higher consumption, and many other multiple factors along the food chain from primary producer to the consumer. Several incidences related to well-recognized foodborne pathogens such as E. coli, Salmonella spp., L. monocytogenes, Clostridium botulinum, S. aureus, and mycotoxin producing fungi species in nonalcoholic beverage products have been reported in many countries, however, current understanding of the trends in foodborne diseases with respect to the nonalcoholic beverages remains largely unknown. Particularly, the awareness and surveillance of viral foodborne pathogens in nonalcoholic beverages seems poorly understood. In order to develop effective prevention and control strategies, more research on novel pathogen detection and control measures is needed. Consequently, the establishment of collaborations among all parties involved in the nonalcoholic beverage industry including researchers, risk assessors and risk managers, public health experts, veterinarians, food safety authorities, and consumers is also necessary.

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Further Reading Kregiel, D., 2015. Health safety of soft drinks: contents, containers, and microorganisms. Biomed. Res. Int. 2015, 15. https://doi.org/10.1155/2015/128697. Lew, L.C., Liong, M.T., 2013. Bioactives from probiotics for dermal health: functions and benefits. J. Appl. Microbiol. 114 (5), 1241–1253. Prado, F.C., Parada, J.L., Pandey, A., Soccol, C.R., 2008. Trends in non-dairy probiotic beverages. Food Res. Int. 41 (2), 111–123. Ranadheera, C.S., Evans, C.A., Adams, M.C., Baines, S.K., 2014b. Effect of dairy probiotic combinations on in vitro gastrointestinal tolerance, intestinal epithelial cell adhesion and cytokine secretion. J. Funct. Foods 8, 18–25.