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Introduction to The Company

We aim to create innovative products along the themes of "natural & healthy" as well as "unique & premium" with our MONOZUKURI based on consumer-centric ideas in each country and region.

SBF's R&D Strength

the research and development team of nature's way beverages

Global Shift in Consumer Preferences toward Natural and Healthy Products

For decades, the growth of the world beverage market was driven by consumer demand for carbonated soft drinks containing sugar. In recent years, however, a growing focus on health and a shift to RTD coffee and tea products have been reflected in increasing consumption of water and non-carbonated soft drinks, which now account for around 70% of the total market. This trend has been especially pronounced in Japan, where the percentage of water and non-carbonated soft drinks has risen to around 80% of the total market. This is even higher than the levels in Europe and North America. SBF believes that this shift in beverage demand will continue, and that consumers will increasingly seek beverages that are further natural and healthy. SBF will respond to these needs by accelerating its development of products that provide enjoyable wellness by combining emotional fulfillment with delicious taste and healthiness.

Research and Development— The Lifeblood of a Manufacturer

To achieve continuing business growth, SBF needs to provide consumers with high-added-value products that explore new needs. Since its founding, the Suntory Group has recognized the importance of research and development as the lifeblood of a manufacturer. This philosophy is reflected in a record of achievement accumulated through decades of R&D activities. Through the process of developing various products, starting with liquor & spirits manufacturing, the Suntory Group has deepened its knowledge of distillation technology and the technologies needed to utilize microorganisms and enzymes. In addition to these technologies, SBF also uses knowledge gained through other types of R&D, such as the exploration of new health-related ingredients, research into health benefits, and water science. We apply these capabilities to create new flavors and ingredients based on natural materials.

Co-creation of Global Value

Our mission is to provide consumers worldwide with satisfying, reliable, and safe products. Since the founding of SBF, our MONOZUKURI, or manufacturing, activities have been guided by the fundamental values and principles of MONOZUKURI that are today shared throughout the SBF Group as the "MONOZUKURI Way." As noted earlier in this report, the Japanese market is leading a trend toward products that are more natural and healthy. SBF has reflected this trend in its own product portfolios, which are "natural and healthy" and "unique and premium." We are confident that we can use our technology development capabilities and expertise in overseas as well as domestic markets. In addition to our continuing efforts to provide new value, we also explore taste experiences that match consumer preferences in various markets. To ensure that our activities closely reflect consumer needs in every region, we apply a consumer-centric approach to all aspects of our global product development activities. The global SBF Group aims to provide consumers around the world with enjoyable wellness through synergies made possible by its many brands, its technologies, and its expertise.

Globalized product development

"We are strengthening mutual understanding and trust throughout the Group with an approach based on local autonomy, with the aim of enhancing global synergies in R&D."

Teruyoshi Morikawa

Senior General Manager Research and Development Department MONOZUKURI Division Suntory Beverage & Food Limited

Role and Function of the Research & Development within the MONOZUKURI Division

Our key role is to support independent product development activities in each region, specifically through cross-organizational activities ranging from talent development to technology development and technical support. This role includes the provision of materials and technologies as the source of new value, technical support for development processes and mass-production, and technical advice about various issues at other stages of product development.

Creating a Unique Global Soft Drink Business Built on Local Autonomy

In April 2017, the Research & Development Department was created within the MONOZUKURI Division through the merger of the R&D Planning Division, New Product Development Division, and Research & Technology Development Division. We have been developing cooperation among the various regions over the past three years, but I sense that the evolution of trust and communication among group companies has accelerated since last year. This is the result of our newfound ability to offer more comprehensive solutions to each region. From a product development perspective, we must provide the support necessary for applications to be adapted to local needs, while maintaining respect for local leadership and ownership. We base our activities on the concept of local autonomy, because we recognize the need to understand regional and national markets when taking products to the commercialization stage.

Recent Achievements Resulting from Global Synergies

One recent success resulting from global synergies is MayTea, which was launched in May 2016 in France, where sugar-related issues have become a focus of controversy. MayTea 's positioning as a low-sugar premium iced tea has allowed it to rise to the number-two position in the French ready-to-drink (RTD) iced tea market last year, which was only the second year since its launch. The development of MayTea was a fundamentally local initiative, but I believe that the Research & Development Department contributed in a number of ways, including evaluation and extraction methods for the tea itself, as well as the assessment of product benefits. goodmood, a mixture of fruit extract and purified water produced using a localized version of a Japanese recipe, was launched in Indonesia in March 2017. In the year since the product launch, goodmood has become increasingly popular in the local market. In Oceania, Frucor Suntory has launched a new line of kombucha beverages named " Amplify ." These new products were developed in Oceania and are being sold on a small scale as the first step. Because these beverages are fermented using a combination of bacteria and yeast, including lactic acid bacteria, microbial control plays a key role in mass production. As Frucor Suntory executes its market expansion plans, the Research & Development Department is providing our expertise and technical support for continual improvement of quality control and the scale up of mass production.

Advantages in the Global Market

SBF's Promise, Mizu To Ikiru, expresses the shared commitment of the entire SBF Group to the protection of the natural environment, and the optimal use of the gifts of nature. Our beverages, which rely on ingredients from nature, offer a variety of benefits. They are delicious and refreshing, and they quench thirst. Some improve concentration, while others help people to relax. We are aware that consumers are highly selective in their beverage decisions, and our first priority is to supply products that are safe and healthy. We want people around the world to know that SBF products are natural, healthy, reliable, and safe, and that our beverages will contribute to their well-being and their enjoyment of life. Trust is our most valuable asset, and we must never betray our customers' trust in us. I believe that such trust will help to establish SBF as a global company.

Developing products to satisfy the needs and preferences of local consumers

Suntory tennensui.

To respond to consumer demands for natural and healthy products

General Manager Development and Design Department Japan Business Division

Norihiko Yoshimoto

Suntory Tennensui mineral water is originally a blessing from nature, and by definition something that can be enjoyed safely and with peace of mind. Our aim in developing the Suntory Tennensui brand is maximize the wonderful benefits of this mineral water and offer high-added-value products to consumers by combining rigorously selected natural ingredients to create new products that will provide great tastes and emotional fulfillment. Natural ingredients have a variety of complex tastes and flavors. By carefully bringing out those tastes and flavors, we can create extremely enjoyable products. However, because these ingredients are natural, it is also extremely difficult to control their compositional consistency. That is the most challenging aspect of our development work. In our R&D activities in Japan, we combine development knowledge accumulated over many years, and our endless commitment to originality and innovation, with science and technology, to create unique new products for the Suntory Tennensui range, such as Asa-zumi Orange & Tennensui, Yogurina & Tennensui, PREMIUM MORNING TEA Lemon, and PREMIUM MORNING TEA Milk . In addition to our efforts to create delicious tastes and flavors, our formula designs are also based on detailed research concerning the mechanisms of the human body. Our aim is to understand why human beings like certain tastes and flavors and want to drink beverages with those. I believe that what we have learned from our R&D work in these areas will increasingly be used in R&D activities throughout the entire SBF Group. Every year, the R&D staff in Japan absorbs new knowledge by exploring consumer trends in other countries. We share that knowledge with our colleagues in overseas SBF Group companies. We will continue to respond to consumer demands worldwide for natural and healthy products by creating new beverages to be enjoyed by people.

Refreshing and natural taste with no artificial flavorings

Director Research and Development Regulatory Affairs and Industrialization (R&D) Orangina Suntory France

Catherine Hubert

SBF Europe's concept of MayTea was in part a response to the general downtrend in the sale of sweet drinks, but also an effort to provide what customers were looking for and could not find on the market. The MayTea infusions are authentic, refreshing and natural, with a short ingredient list and no artificial flavorings, colors, or preservatives, and are also very low in sugar. SBF Japan's R&D helped the local team throughout the developmental process, especially in the area of tea infusion technology, by offering advice on key points like water quality. In addition to flavor adjustments to the French consumers' taste, the R&D teams faced the issue of the product's cloudiness, as particles from the infusion would settle at the bottom of the bottle. Solving that issue was one of the greatest challenges they overcame. While SBF Europe's R&D received a productive boost from working with Japan R&D, the product remained local consumer oriented. Essentially, SBF gives local companies control of how they want to use the technological platforms that have been developed. For example, one of the first two flavors that the local team developed was a green tea and mint infusion. This was created specifically to offer an appealing flavor to a large number of French customers familiar with Moroccan and North African tea culture. MayTea 's sales results seem to be solid evidence for the effectiveness of that management style. In two years' time, MayTea became the second highest-selling product in the French bottled iced tea market, creating the new category of infused tea, and tripling its sales numbers in three years. After its debut in France, the brand went on sale in Belgium and Spain, with plans to expand in other European countries in the future.

To offer something new to those looking to quench their thirst

Vice President Research and Development Suntory Garuda Group

Yasuhiro Yamanishi

Senior Manager Research and Development Suntory Garuda Group

Niken Sulastri

With temperatures hovering at tropical levels all year round, it is perhaps unsurprising that the Indonesian beverage market is made up of nearly 50% bottled water. In order to offer something new to those looking to quench their thirst, Suntory Garuda developed goodmood , a clear, fruit infused water. With the product's release in the spring of 2017, the company became a pioneer in the fruit infused water category in Indonesia. Starting with the same core technology as the SBF Japan's R&D team, but adapting and recreating the recipe to fit a new market and the local Indonesian palate became a chance for unique product development. Indonesian consumers have reacted positively to the concept of infused water, which looks healthy and has an appealing natural taste. We consulted SBF Japan's R&D department on challenges like creating natural fruit taste, while also learning about the benefits of osmotic pressure for hydration. The R&D teams faced some challenges but succeeded in capturing the right level of sweetness and acidic citrus taste that Indonesian consumers preferred. The recipe also had to be adjusted to be suitable for the Halal standards of the market. Suntory Garuda launched two flavors: honey lemon and orange. Because Indonesian consumers love sweet citrus, we added honey to the lemon, and chose a sweet orange taste profile. While the size of the Indonesian beverage market is still small, SBF sees it as a future growth area. Our R&D team is already considering other ways in which they can benefit from SBF's water and hydration know-how to create more innovative and attractive products for their consumers.

To explore a completely new product area, kombucha

Technical Manager Australia Frucor Suntory (Australia) Pty Ltd

Ben Walkley

After seeing consistent decreased sales of juice drinks as many consumers cut back on their sugar intake, Frucor Suntory decided to explore a completely new product area: kombucha. The fizzy, fermented, non-alcohol tea beverage is one recently trending alternative to sugary beverages. Kombucha's wide range of potential health benefits and a reliance on natural ingredients also made it a good fit for the SBF Group portfolio. When Frucor Suntory was looking into developing kombucha, there were already local brewers selling their own versions. The small size of these brewers, however, meant there was often a fluctuation in the flavour and quality of the finished drink. Frucor Suntory wanted to create a product that could provide both consistent quality and a less acidic taste for consumers. The Frucor Suntory R&D team continually finessed the recipe over the course of development to get it just the way they wanted. Kombucha requires fermentation for many days, which is very different to the other products the company manufactures. Currently, the R&D team is interested in deepening their understanding of the SCOBY, the symbiotic culture of bacteria and yeast, which creates the distinctive taste of kombucha. They will call on the technical assistance of SBF Japan's R&D for the necessary scientific analysis. The kombucha, under the brand name Amplify , is certainly one of the more adventurous products that an SBF Group company has released in the past couple of years. Three flavours of Amplify went on sale in Australia in January 2018, and in New Zealand in April 2018. Though it is too early to say whether this will be the start of a strong new segment, there does seem to be a groundswell of interest. By successfully bringing Amplify to market, Frucor Suntory's R&D department has also gained the insight needed to potentially assist other SBF Group R&D teams who may be interested in producing kombucha in the future.

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Evolution of Food Fermentation Processes and the Use of Multi-Omics in Deciphering the Roles of the Microbiota

Mohamed mannaa.

1 Department of Integrated Biological Science, Pusan National University, Busan 46241, Korea; ge.ude.uc@aannam (M.M.); rk.ca.nasup@enoorc (G.H.)

2 Department of Plant Pathology, Cairo University, Giza 12613, Egypt

Young-Su Seo

Inmyoung park.

3 School of Culinary Arts, Youngsan University, Busan 48015, Korea

Food fermentation has been practised since ancient times to improve sensory properties and food preservation. This review discusses the process of fermentation, which has undergone remarkable improvement over the years, from relying on natural microbes and spontaneous fermentation to back-slopping and the use of starter cultures. Modern biotechnological approaches, including genome editing using CRISPR/Cas9, have been investigated and hold promise for improving the fermentation process. The invention of next-generation sequencing techniques and the rise of meta-omics tools have advanced our knowledge on the characterisation of microbiomes involved in food fermentation and their functional roles. The contribution and potential advantages of meta-omics technologies in understanding the process of fermentation and examples of recent studies utilising multi-omics approaches for studying food-fermentation microbiomes are reviewed. Recent technological advances in studying food fermentation have provided insights into the ancient wisdom in the practice of food fermentation, such as the choice of substrates and fermentation conditions leading to desirable properties. This review aims to stimulate research on the process of fermentation and the associated microbiomes to produce fermented food efficiently and sustainably. Prospects and the usefulness of recent advances in molecular tools and integrated multi-omics approaches are highlighted.

1. Introduction

The art of fermentation is as old as the human civilisation on earth, as it was traditionally developed by ancient societies for food preservation during harsh seasons, for ritual feasts, and to enhance the sensory quality of food [ 1 ]. Historical records reveal that fermentation of several substrates, including milk and cereal, is indigenous to many parts of the world. The earliest form of fermentation was discovered by analysing the stone mortars from the Natufian burial sites of a semi-sedentary foraging population, providing archaeological evidence for beer brewing from cereals dating back 13,000 years [ 2 ]. In ancient Egypt, dairy products, fermented bread, and beer were dietary staples [ 3 ]. In China, chemical analysis of ancient pottery jars indicate the existence of fermented products of rice, honey, and fruits as early as the seventh millennium B.C. [ 4 ].

Fermentation was an integral part of other ancient civilisations; examples include beer brewing in Babylonia, soy sauce production in East Asia, and fruit fermentation in Greece (Greeks attributed ‘Dionysos’ as the god of fruit fermentation) [ 5 ]. The most ubiquitous type of fermented food is yoghurt (made from milk), which was produced and consumed throughout the Middle East and Europe and has become a major component of the human diet worldwide [ 6 ]. In East Asia, a series of fermented food products, mainly based on rice, soybean, vegetables, and fish, have been developed and are still produced and consumed on a daily basis; examples of such products include Korean kimchi and Japanese natto, which have gained popularity worldwide owing to their unique taste and proven health benefits [ 7 ].

Fermentation continues to be practised due to the evidence of extended shelf life and improved organoleptic properties of fermented foods. There is a wide variation in fermented foods and drinks prepared and consumed worldwide, although we lack detailed knowledge of the microbial properties underlying such variation [ 5 ]. In the past, fermentation was based on naturally occurring microbes in the food substrate that were affected largely by the surrounding condition and environment, leading to the characteristics of the fermented products according to their geographical location. With the growing global attention and the increasing demand for fermented products, merged with the increased awareness of food safety aspects, standardisation of the process was necessary, which led to industrial control of the production procedures, such as the use of starter cultures and the control of fermentation protocols on an industrial scale [ 8 ].

The first model for understanding microbial roles in food fermentation was created when the role of the fungus Aspergillus oryzae in the preparation of koji was discovered by the German scientist Korschelt in 1878 [ 9 ]. This discovery was followed by the identification of various additional fermentation microbes and starter cultures, reaching the recent advancement in molecular biology techniques, next-generation sequencing (NGS), multi-omics and bioinformatics tools, and advanced statistical approaches. These advancements have revealed the microbiome composition of fermented foods and the complete genome sequence of biotechnologically important microbes. These findings have enabled a thorough understanding of the fermentation process and microbial diversity, roles, and metabolic pathways and outcomes, leading to great development in the fermentation industry [ 9 ].

Human gut microbiome research has revealed the link between the gut microbiome and different aspects of human health and diseases. This finding has necessitated studies on fermented foods and their roles in enhancing the microbiome. This review aims to discuss the evolution of the fermentation process and the contribution of modern biotechnological tools in improving the process of food fermentation. Furthermore, the modern advancements in multi-omics approaches are reviewed along with their application in studying the fermentation-associated microbiome, microbial interactions within the fermented food ecosystem, and microbial roles in imparting fermented foods and beverages with unique properties. Finally, the future perspectives on food fermentation considering modern innovative approaches are highlighted.

2. Classification of Major Types of Fermented Foods and Beverages

Fermentation involves the action of enzymes and catalysts derived from microorganisms such as bacteria, yeast, and moulds for the chemical transformation of the complex organic compounds in the substrate into simpler, bioactive, functional, and nutritious compounds [ 10 ]. There are several classification methods for fermented foods and beverages, mainly based on the substrate category used, such as fermented milk, cereal, legumes, vegetables, fruits, meat, fish, and herbs [ 11 ]. The various combinations of different types of food substrates and the involved fermentation microbiota give rise to thousands of fermented products worldwide; the main examples of common fermented foods and beverages and the main fermenting microbes are summarised in Figure 1 .

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Object name is foods-10-02861-g001.jpg

Common fermentation substrates and produced fermented foods and beverages.

Fermentation can also be categorised, according to the main biochemical pathway, into four basic categories: alcoholic, lactic, acetic, and alkali fermentation ( Table 1 ) [ 12 ]. In alcoholic fermentation, the sugars in the substrate are converted into alcohol and carbon dioxide; examples of such fermentation include the production of bread, beer, and wine. Yeast is the predominant microbe responsible for this type of fermentation. In lactic fermentation, sugars are converted into lactic acid, as in the case of yoghurt, kimchi, and fermented cereals. Lactic acid bacteria (LAB) are mainly responsible for this type of fermentation [ 13 ]. In acetic fermentation, organic compounds such as alcohols and sugars in the substrate are converted into acetic acid by bacteria mainly belonging to the genus Acetobacter , as in the case of production of water kefir, kombucha, cocoa, acidic beer, and vinegar [ 14 ]. Organic acids that are microbially produced during the fermentation of several fermented foods and beverages play key roles in determining the quality and safety aspects of the products. For example, propionic acid, produced by Propionibacterium , and glucuronic acid, produced mainly by Gluconacetobacter , impart kombucha with antioxidant properties and strong antimicrobial activity against harmful microbes [ 15 ]. The lactic acid bacteria could also be classified into two main physiological groups depending on the fermentation pathway, the homofermentative and heterofermentative. The distinction is in the main product of the fermentation of sugars being primarily lactic acid in the homofermentative and lactic acid, CO 2 , acetic acid and/or ethanol in the heterofermentative group [ 16 ].

Classification of the major types of fermentation related to food production.

In alkali fermentation, the proteins in the substate are hydrolysed into amino acids and peptides, releasing ammonia, which elevates the pH (8–9), inhibiting spoilage-associated microbes. Ammonia produced during alkaline fermentation (involved in the preparation of Japanese nattu and African fermented legumes and eggs) is responsible for a strong umami flavour and aroma. Microbes responsible for alkaline fermentation mainly belong to Bacillus spp. and coagulase-negative Staphylococcus , which can produce extracellular proteinase for protein hydrolysis [ 17 , 18 ]. Unlike fermented products that depend on a specific group of microbes for fermentation, there are fermented foods and beverages, such as Korean doenjang and kombucha, that pass through different stages of fermentation, in which different types of microbes are responsible for the multi-step fermentation process [ 19 ].

Steinkraus [ 20 ] proposed a seven-category classification of fermented foods and beverages that predicts the involved microorganisms and the changes (chemical, physical, and nutritive) occurring during fermentation. In this classification, textured vegetable-protein meat substitutes such as Indonesian tempe, high salt/meat-flavoured amino acid/peptide sauces, fermented paste such as fish sauce and miso, and leavened and sourdough breads were added as separate categories to the previously described general classification [ 20 ].

3. Evolution of the Process of Fermentation over the Years

3.1. spontaneous fermentation and back-slopping.

Fermented foods and beverages have traditionally been produced by relying on the microbiota naturally occurring on the food substrate. Spontaneous fermentation dependent on autochthonous microbes was the main method for producing fermented food and beverages throughout history, and it remains a mainstay method in domestic, small-scale, and household settings [ 21 ]. In this type of fermentation, the conditions are adjusted to allow for the growth of desirable fermentation microbes that impart unique sensory properties to the product and prevent the growth of spoilage-associated microbes [ 22 ]. Fermentation conditions often need to be adjusted—for example, creating anaerobic conditions is necessary for the production of pickles; the composition of ingredients may also need to be adjusted (e.g., by adding salt or vinegar during fermentation) to suppress competing undesirable microflora [ 23 ]. Many types of fermented foods, such as sauerkraut and kimchi, are still produced using spontaneous approaches without the use of starter cultures, especially in small-scale settings and in developing countries, as the process depends completely on enhancing the growth of microbes available on the substrate raw materials [ 12 ].

The start of the fermentation process may involve transferring a small amount of a previously successful fermented batch into fresh ingredients as an inoculum to facilitate the initial phase of fermentation of the next batch, even without the knowledge of the types of active microbes; this process is called back-slopping [ 24 ]. In spontaneous fermentation, a successful process is achieved when the desirable microbes can outcompete and dominate harmful and spoilage-associated microbes because of their adaptability to the substrate and the prevailing fermentation condition. Back-slopping reduces the risk of failure and facilitates the competitive ability of fermentation microbes; repeating the process provides further selection of useful microbes that are best adapted to the food substrate and the fermentation condition, providing the currently available starter cultures [ 22 ].

3.2. Starter Cultures

With progress in the microbiological techniques of isolation, identification, and microbial preservation, specific starter cultures have been isolated and characterised from fermented foods. These cultures are currently used, especially on the industrial scale, to ensure that the process is controlled and the fermentation outcome is stable for quality and properties. The use of well-defined starter cultures was first adopted to produce beer, alcohol, vinegar, and bread, followed by dairy and meat products [ 25 ]. The main role of starter cultures is to accelerate the fermentation process and to convert carbohydrates in the substrate into alcohols and organic acids which act as natural preservatives that restrict the growth of harmful microbes and impart distinct and desirable organoleptic properties to the product. Minimising the risk of foodborne diseases has been confirmed previously in natural conditions and in artificial inoculation with pathogens [ 26 , 27 ]. The release of carbon dioxide by the action of starter cultures is important for the process of fermentation, as it contributes to rising the dough during breadmaking, making the foam of beer and buttermilk, and the formation of eyes in cheese [ 25 ].

The starter cultures that are mostly used to produce fermented foods and beverages, particularly acidic fermented products, belong to LAB [ 25 ]. Such bacteria include members of Lactobacillus, Leuconostoc , Enterococcus, Streptococcus, Oenococcus , and Pediococcus , and some of them may exert direct beneficial effects on health as live probiotic microbes [ 28 ]. Additionally, non-LAB bacteria, such as those belonging to Bacillus , Micrococcaceae , Bifidobacterium , Propionibacterium , and Brachybacterium , act as a secondary group of microorganisms in the fermentation process [ 29 ]. Along with bacteria, yeast and moulds, including several species of Debaryomyces, Kluyveromyces, Saccharomyces, Aspergillus, Mucor, Penicillium, and Rhizopus species, represent an important part of starter cultures in a variety of fermented foods and beverages, such as cheese and coffee, in which the microbiota significantly affects the appearance and organoleptic properties [ 30 , 31 ]. For wine making, the Saccharomyces cerevisiae is traditionally used in the fermentation process. In addition, there is an increasing awareness about the enological characteristics of other non- Saccharomyces yeast in imparting the wine with particular flavour and aroma [ 32 ].

3.3. Starter Cultures of Multiple Strains and Adaptation for Co-Existence

Starter cultures do not always contain a single strain; in many cases, a consortium of different organisms and strains is involved. The model example for the fermented beverages to be covered in this review is kombucha, with a starter culture consortium of multiple species that are well-adapted to co-existence. In the case of kombucha, a fermented, sweetened, black tea-derived beverage, which originated in China thousands of years ago and is currently gaining popularity worldwide for its health-promoting and therapeutic effects, a symbiotic culture of bacteria and yeast (SCOBY) is used to initiate fermentation [ 33 ]. Kombucha fermentation comprises three main types of fermentation (i.e., alcoholic, lactic, and acetic) due to the presence of different types of bacteria and yeast co-existing in the medium and responsible for different stages of fermentation; the process is initiated by osmotolerant microbes, and acid-tolerant bacteria prevail and dominate [ 34 ].

In this case, the microbes are well-adapted to the substrate and co-exist with other microbes constituting the SCOBY. They act in harmony; the substrate contains sucrose that is first broken down by the action of yeast ( Saccharomyces cerevisiae ) into fructose and glucose, which are then used for the growth of bacteria in the consortium (e.g., Acetobacter and Gluconobacter spp.) producing various organic acids, such as acetic, gluconic, and glucuronic acids [ 35 ]. Yeast in kombucha ferments the sugar into ethanol and CO 2 ; ethanol is subsequently oxidised into acetic acid by acetic acid bacteria. These organic acids, along with the alcohols produced by the yeast, act as antimicrobial agents that inhibit the growth of undesirable microbes in kombucha [ 35 ]. The levels of polyphenols and flavonoids originally found in black tea increase progressively with fermentation, most likely due to the role of yeast in enzymatically degrading the polyphenols into smaller molecules, increasing the antioxidant activity of kombucha and stimulating the production of bacterial cellulose [ 36 , 37 ].

The microbial cellulose produced by Komagataeibacter xylinus (formerly Gluconacetobacter xylinus ) is the base for forming the floating biofilm as the solid phase of kombucha. Formation of this biofilm enhances the association between bacteria and yeast and plays a role in adjusting the fermentation condition to support the survival of important microbial groups by retaining bacteria and yeast on the surface of the liquid to ensure adequate oxygen supply and nutrient diffusion (by forming reticulation) to inhabiting bacteria [ 38 ]. The roles of the microbial agents within the kombucha ecosystem are not limited to their biological activity; even after the death of the involved yeast cells, they release vitamins and nutrients, stimulating the growth of important bacteria [ 34 ]. This phenomenal co-existence of different interacting microbes constituting the consortium of kombucha fermentation represents a model for fermentation microbiota co-evolution, powerful symbiosis, and ecological system stability; this consortium can tolerate simulated Mars-like environmental conditions and restore their biological activity after exposure [ 34 , 39 ]. Figure 2 illustrates the metabolic interplay and functional compatibility of the kombucha fermentation microbes, indicating their adaptation and strong symbiosis.

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An illustration of the metabolic interplay and functional compatibility of kombucha fermentation microbiota, representing a model for the adaptation and symbiosis of the microbiota in the fermentation ecosystem.

3.4. Genetic Improvement of Starter Cultures

The fermentation process has further evolved as the use of starter cultures has undergone significant improvement with the advancement of molecular biology techniques. Previously, the selection of starter cultures was based on the screening of many isolates, and those that performed well in fermentation on an industrial scale, yielding end products with acceptable organoleptic characteristics, were selected [ 40 ]. Recently, advanced tools allowing for high-throughput screening for specific targeting of genes and metabolic pathways have resulted in the selection of better performing and well-adapted starter cultures for improved fermentation and facilitated the selection of mutants and genetic engineering for superior starter cultures with desired properties [ 25 ].

The successful plasmid transformation of Lactococcus lactis (formerly Streptococcus lactis ), an important microorganism for dairy fermentation, using recombinant DNA techniques in 1982, was considered a turning point for using genetic engineering to improve starter cultures for preparing fermented food [ 41 ]. Following this advancement, several industrially important LAB, such as Streptococcus thermophilus and members of the Leuconostoc genus, have been genetic modified to improve traits linked to metabolism, efficiency of proteolysis, and defence against bacteriophages [ 42 ]. Infection of starter cultures with bacteriophages is a major concern for dairy fermentation, as this causes significant economic losses due to the rapid accumulation of bacteriophages, leading to the complete termination of acidification and consequent spoilage [ 43 ]. Progress in molecular biology has led to the characterisation of bacteriophages coupled with sequencing of the whole genome of L. lactis , facilitating understanding of the process of bacteriophage infection and bacterial defence mechanisms. This discovery was translated into constructing strains with bacteriophage components that inhibited phage proliferation and offered significant protection to L. lactis [ 44 ]. Moreover, genetic and metabolic engineering of LAB opens the way for further utilisation of milk lactose during fermentation with the possibility of generating new useful products (both simple and complex) along with lactic acid, with various beneficial applications [ 45 ].

Starter cultures have been generated using recombinant DNA technology for decades and may provide improved fermentation processes and offer better-quality products with desired properties. Despite such potential, none of the developed strains are being used in the industry due to strict governmental regulations and the lack of consumer acceptance of genetically modified food ingredients [ 46 ]. Therefore, strain improvement methods without the use of recombinant DNA technology, such as random mutagenesis, directed or adaptive evolution, and dominant selection, together with natural mechanisms such as bacteriophage transduction, natural competence, and conjugation, are widely used in the food industry [ 47 ]. Random mutagenesis induced by classical methods (e.g., UV treatment) and subsequent selection of useful variants have been successfully used to generate starter cultures with desired properties [ 48 , 49 ]. However, this method has the disadvantage of causing unintended mutations that might impair the applicability of strains and affect their performance, as evidenced by bacterial whole genome studies [ 47 ]. Improved strains qualify as ‘generally regarded as safe’ by the U.S. FDA if they have genetic stability, no foreign DNA, no antibiotic-resistance-marker genes, and no global changes from the parental strain; such improved strains have been generated and registered for use [ 50 ]. One example of a registered improved strain as starter culture is the metabolically engineered urea-degrading Saccharomyces cerevisiae yeast strain generated to reduce the content of ethyl carbamate, a potent carcinogen in wine; the strain reduces ethyl carbamate in wine by 89.1% and was patented and registered for producing alcoholic beverages [ 51 ].

3.5. CRISPR/Cas9 Technology for Genetic Improvement of Starter Cultures

The revolutionary novel technology, CRISPR/Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated protein 9), is an extremely precise method of gene editing; it has taken genetic engineering to another level with a wide range of biotechnological applications in many fields, and its discoverers were selected for the Nobel Prize in Chemistry in 2020 [ 52 , 53 ]. Briefly, CRISPR/Cas9 is based on the mechanism of ‘adaptive immunity’, which is naturally found in bacteria and archaea, and comprises two components: the chimeric guide RNA (gRNA) and the RNA-guided DNA endonuclease (e.g., Cas9); the CRISPR/Cas9 toolbox can be used for precise genome editing of any organism [ 54 ]. The applications of the toolbox in the food industry are numerous, and it has been applied to improve the strains of starter cultures by producing marker-less, genetically stable strains with improved properties [ 50 ].

The technology was first applied in 2013 for genome engineering of Saccharomyces cerevisiae, an industrially important yeast and starter strain for several fermented products [ 55 ]. Since then, several applications of the CRISPR/Cas9 toolbox to improve the applicability of S. cerevisiae have followed. Recently, engineering yeast for the reduction of production of urea, the precursor of ethyl carbamate, and the modulation of glycerol production in wine have been successfully implemented [ 56 , 57 , 58 ]. The application of the CRISPR/Cas9 toolbox has been explored for improving fermented food and beverage starter microbes along with yeast. Kimchi-associated Leuconostoc citreum was engineered using the CRISPR/Cas9 toolbox for elimination of cryptic plasmids and this process was suggested as a food-grade method to develop a safe lactic acid bacterial strain without residual antibiotic markers [ 59 ]. Katayama et al. [ 60 ] utilised the CRISPR/Cas9 toolbox to develop a functional and versatile genome editing method for efficiently targeting mutagenesis in A. oryzae , which is an industrially important filamentous fungus used in Japanese and Korean traditional fermentation [ 60 ].

The novel CRISPR technology for gene editing is promising for food-grade applications; it is highly precise, stable, and should be considered outside the scope of genetically modified organisms (GMO) as the modification occurs in nature [ 50 ]. However, it still falls into the definition of GMO according to the European Union court, as it concerns organisms made through in vitro mutagenesis [ 61 ]. The scientific community is putting effort into the reconsideration of such regulations and the acceptance of the technology in the food industry [ 62 ]. Nevertheless, research on improving the properties of starter cultures of fermented food and beverages is critical to improve the quality of products and reduce the potential hazards posed by undesired microbes or their metabolites.

3.6. CRISPR-Mediated Microbiome Engineering and Fermentation

Microbial activity in fermented foods and beverages is the main factor responsible for product quality and safety. Hence, the manipulation of the microbial composition, particularly at the initiation of fermentation, is key for controlling the process and shaping the properties of the product. Recent advances in biotechnological tools and the rise of CRISPR-based technologies may not only be involved in genetic improvement of specific strains in starter cultures, as explained earlier, but the technology also has great potential in microbiome engineering [ 63 ]. This superior method can target specific groups of undesirable microbes within the fermented food ecosystem and control the microbiota assembly to enhance desirable fermentation microbes, leading to the optimisation of fermented food products [ 64 ].

CRISPR-mediated microbiome manipulation has been investigated in several recent studies. The main priority for fermented food and beverage microbiome manipulation is to selectively control the undesirable spoilage-associated microbes or microbes competing with fermentation-desirable microbes to enhance product quality and extend shelf life [ 64 ]. Specific targeting of specific individual microbial strains within microbial consortia was previously carried out using the CRISPR/Cas9 toolbox in Escherichia coli as a model microbe by targeting specific sequences [ 65 ].

Such specific targeting of individual strains in mixed cultures and the differentiation between pathogenic and beneficial microbes was nearly impossible using tailored growth conditions or traditional antibiotics; such targeting allows for further applications by selectively clearing contaminating microorganisms and quantitively controlling the environmental or industrial microbial community composition [ 65 ]. The specific DNA sequences responsible for undesirable features that are unique to pathogenic or spoilage microbes, such as virulence factors, antibiotic resistance, or toxin production, can be targeted for elimination [ 64 , 66 , 67 ]. Although most studies utilising innovative CRISPR-based selective antimicrobial approaches focus on pathogenic microbes, involving selective manipulation of the food-fermentation microbiome by targeting the genotype of spoilage microbes. The different applications and contributions of CRISPR-based technologies in improving food fermentation by targeted gene editing for improvement of starter culture microbiome engineering are shown in Figure 3 .

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CRISPR-Cas9-mediated gene editing and its possible applications in food fermentation. The precise gene editing can be utilised for improving of the starter culture by deletion of undesirable traits or insertion of desirable traits. CRISPR-Cas9 technology could be utilised for microbiome engineering by targeting unique sequences and selectively eliminate spoilage and undesirable microbes from the community.

CRISPR-based microbiome engineering represents the most recent advancement in the evolution of the food fermentation process, starting with traditional spontaneous fermentation through the use of starter cultures and genetic engineering. The evolution of the food fermentation process is summarised in Figure 4 .

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Evolution of the process of fermentation throughout the history, starting from relying on the natural indigenous microbes reaching to the innovative approach of microbiome engineering using advanced technological tools.

4. Multi-Omics and Microbiota Dynamics of Food Fermentation

The continuously interacting microbiota of food fermentation ecosystems, encompassing different types of bacteria, yeast, and fungi, plays a major role in shaping the quality and safety of fermented foods and beverages [ 68 ]. Previous studies investigating the microbial composition of fermented products were based on the culture-based traditional plate cultivation method, which failed to provide accurate information about the microbial profiles mainly because of the vast majority of uncultivatable microbes and the presence of viable but not culturable microbes, especially in fermented food ecosystems [ 21 , 69 ]. The recent advancement of the NGS technology, incorporating the collective studies of microbial genomes and metagenomics, meta-transcriptomics, meta-proteomics, and metabolomics to study microbial communities, has enabled accurate identification of the microbial composition of different ecosystems, including fermented food, and the detailed study of the microbe-microbe and microbe-environment interactions within food fermentation ecosystems, involving microbial gene expression, activities, and metabolomic interplay [ 70 ]. These nucleic acid and protein-based next-generation approaches have replaced traditional culture-based methods for microbial community profiling and are the cornerstone for understanding the fermentation process in detail and providing opportunities to interfere and manipulate the community composition for improved fermentation processes for safer and better-quality products with desired properties and extended shelf life [ 71 ]. Integrated multi-omics analyses and their roles in studying the fermentation microbial ecosystem are summarised in Figure 5 .

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The different tools of the multi-omics analysis and their roles in understanding the food fermentation process, microbiome structure and functional activity profiling.

One of the most extensively applied techniques in recent studies to profile the microbiota within food fermentation is the use of high-throughput sequencing (HTS)-based metabarcoding, by analysing the collective genomic markers by employing universal primers, such as the 16S rRNA and internal transcribed spacer (ITS) regions for bacteria and fungi, respectively [ 71 ]. The limitation of this method is that it might fail to identify the microbes involved at the species level, although in many cases the species could be inferred due to the limited number of identified species within many food fermentation genera [ 71 ]. Furthermore, this method can only provide qualitative and pseudo-quantitative assessment of the present microbiota that would be expressed as ‘relative abundance’ and this limitation could be overcome by integrating targeted molecular cell enumeration techniques to provide an absolute abundance assessment [ 72 ]. Relying on the amplification from DNA templates may limit evaluating the actual active microbial groups, which could be avoided by using RNA-based approaches, including the reverse-transcription of mixed RNA followed by amplification of cDNA that can profile active microbial populations and quantitative PCR for microbial enumeration [ 73 ].

A more comprehensive approach to study the fermented food microbiome is the application of metagenomic shotgun DNA-seq, which provides more accurate taxonomic information on the microbial communities of high-complexity samples, allowing for profiling the functional potential by detection of the global gene content and identification of unknown species [ 74 ]. The applications of the shotgun DNA-seq approach in studying food matrices include the detection of foodborne pathogens and monitoring changes in the gene content during the fermentation process [ 71 , 75 , 76 , 77 ]. Nevertheless, the HTS-based metabarcoding method provides a powerful tool to profile the microbiota of food fermentation and has been successfully applied to investigate different types of food fermentation products and to monitor the microbial dynamics and possible alterations in the microbiota by adjusting the external perturbations such as fermentation conditions, ingredients, and sampling points [ 78 ]. A comprehensive review of studies utilising amplicon based HTS was reviewed by Ferrocino and Cocolin, 2017 [ 71 ]. Among the applications of the HTS-based metabarcoding approach, the microbiota involved in the process of fermentation and the influence of manipulating the fermentation ingredients on the microbial community structure have been investigated.

4.1. Examples on the Use of Meta-Omics to Study Microbial Dynamics of Food Fermentation in Recent Studies

Among the model examples for studies involving HTS-based metabarcoding approach to profile the microbial community structure and evaluate the influence of fermentation ingredients on the microbial community composition and functional potential that will be covered in this review are kimchi, fermented soy products, and kombucha. Kimchi is the most famous Korean fermented food that has gained worldwide popularity owing to its health-promoting properties [ 79 ]. Several studies have investigated the origins of the microbial community and their dynamics in kimchi, as well as the identity of main microbes involved in the fermentation process [ 80 , 81 ]. The main microbes of kimchi fermentation are LAB that initiate fermentation by metabolising vegetable sugars into lactic acid, which reduces the pH and limits the growth of most microbes [ 82 ]. The dominant microbes in kimchi are members of the Leuconostoc , Lactobacillus , and Weissella genera. During the initial fermentation stage, Leuconostoc mesenteroides usually dominates the microbial community, and Lactobacillus sakei and Weissella koreensis begin to dominate at the optimum-ripening and over-ripening fermentation stages; although the latter are important for kimchi fermentation, their rapid growth and activity promote acidic deterioration, decrease the fresh flavour, and reduce the shelf life [ 82 , 83 , 84 , 85 ].

Mannaa et al. [ 85 ] utilised HTS-based metabarcoding to investigate the influence of incorporating gizzard shad fish ( Konosirus punctatus ) during kimchi fermentation on the microbial and chemical composition. The purpose of this study was to evaluate a practice that is traditionally adopted in the coastal cities of Korea, where fish is added during kimchi fermentation, and this type of kimchi has a refreshing taste and a relatively extended shelf life [ 86 , 87 ]. This study revealed that adding gizzard shad fish during kimchi fermentation had a positive effect on the composition of chemicals and the microbiota by reducing the growth of Lactobacillus sakei, which is linked to the rapid acidic deterioration of kimchi, and by promoting the growth of Leuconostoc rapi, which is known for its health-promoting and taste-improving properties owing to the production of the antioxidant mannitol, contributing to the refreshing flavour and desirable characteristics of kimchi [ 85 , 88 , 89 ]. These results shed light on the inherited wisdom in preparing traditional fermented foods by combining specific substrates that provide and enhance the growth of desirable microbes and suppress spoilage. The formulated ingredients in kimchi act as a source of specific microbes and may exert a selective action on the desired microbes by their potential antibiotic effect (e.g., garlic and red pepper powder) against certain microbial species and by adjusting the physicochemical conditions of the fermentation ecosystem [ 90 , 91 , 92 ]. Understanding the fermentation process and adjusting the optimum conditions require investigating the roles of fermentation ingredients in shaping the fermentation microbiome.

In other common Korean fermented products, Korean fermented soy paste (doenjang) and soy sauce (gangjang), manipulating the ingredients during fermentation can cause significant changes in the microbial composition and lead to changes in the properties of the product. Fermentation of doenjang and gangjang is based on two steps, starting with meju, whereby dried soybean blocks are fermented spontaneously using naturally occurring populations of fungi and bacteria. The fermented dry mouldy blocks are then subjected to a second long-term fermentation to produce the solid paste, doenjang, and liquid gangjang [ 93 ]. The process of fermentation is mostly spontaneous and carried out under non-sterile conditions based on natural microbes from the substrates used. Therefore, there is wide variation in the microbial composition of the product, and the substrates play a major role in controlling the taste, safety, and quality aspects [ 94 ].

Mannaa et al. [ 95 ] investigated the influence of incorporating fresh coriander during fermentation to produce gangjang using an HTS-based 16S rRNA metabarcoding and reported significant shifts in the microbial composition compared to the control group. Adding coriander resulted in a significant reduction (~45% reduction) in the relative abundance of Chromohalobacter beijerinckii , which dominated the microbial community in the control group. Reduction in C. beijerinckii is considered beneficial for gangjang as it is responsible for the increase in the levels of biogenic amines, such as histamine, putrescine, and tyramine, which are considered potential health risk factors in fermented salty products and should be minimised. This study combined the metabolomic analysis using 1 H-NMR to evaluate the content of biogenic amines produced in the gangjang, which revealed a significant reduction in the levels of these biogenic amines. This study demonstrates the advantage of integrating multi-omics tools to evaluate the effect of ingredients on the fermentation process and the end product, by suggesting that adding coriander during fermentation has a positive influence on the quality and health of the product.

Similarly, the effect of adding different types of herbs during fermentation for the production of doenjang was investigated using HTS-based 16S rRNA metabarcoding and metabolomic studies [ 96 , 97 ]. The results indicated that the incorporation of herbs, especially peppermint and Korean mint, during doenjang fermentation had a positive effect on the microbial community structure as the levels of undesirable microbes, such as Sphingobacterium and Pantoea , were significantly reduced, while potentially beneficial bioactive metabolite-producing microbes, such as Saccharopolyspora and Buttiauxella , were present at significantly higher levels [ 97 ]. These results were further confirmed and were consistent with those of a recent study that utilised primary and secondary metabolome analyses to evaluate the effect of adding herbs on doenjang properties. The results indicated that adding herbs caused significant shifts in the metabolic composition of both the primary and secondary metabolites, with a more profound positive effect on the secondary metabolites, particularly with peppermint and Korean mint treatments; the levels of isoflavones, soyasaponins, and lysophospholipids were significantly increased along with significantly higher antioxidant capacity compared to doenjang made without herbs [ 96 ].

Metagenomic approaches have been applied to study the microbial composition of kombucha by combining whole metagenome sequencing, 16S rRNA and internal transcribed spacer-1 amplicon analysis. The results indicated that Komagataeibacter and Zygosaccharomyces were dominant at different fermentation times. Moreover, functional complementarity was observed between both microbial groups which explains the sustainability of the kombucha ecosystem by ensuring microbial metabolic cross talks [ 98 ]. Such mutualistic metabolic interplay (briefly described above) between the microbial groups comprising of the kombucha consortia may explain their stability and ability to tolerate harsh environments. The ecological resilience of the kombucha microbiota to long-term exposure to the extremely harsh conditions of the Mars-like conditions in a low Earth orbit was confirmed by shotgun metagenomic analysis as the core microbial structure was maintained, and there were no significant changes in the community functions, such as the ability to produce cellulose-based pellicles, allowing for the survival of the microbial community under extra-terrestrial conditions [ 99 ].

4.2. Functional Activity of the Food Fermentation Microbiome

Several studies have combined meta-transcriptomic analysis, which targets actively expressed genes under specific conditions, with meta-barcoding to decipher the core functions of the detected microbiota that are associated with metabolomic changes affecting the quality properties of fermented foods [ 100 ]. The two approaches were combined to study the structure and function of the core microbiota in Chinese soy sauce aroma type liquor production and facilitate understanding of the flavour development in the product as a two-stage process involving yeast initially for the production of ethanol, followed by a functional shift for the production of organic acids by the action of Lactobacillus [ 101 ]. De Filippis et al. [ 102 ] used a combinatorial approach involving meta-transcriptomic analysis to facilitate understanding of the ripening process of Italian cheese and possibilities for ripening acceleration. The obtained results indicated the roles of non-starter LAB in ripening-related activities and the temperature increase-related modulations on the microbiota structure and function during maturation for optimisation of production efficiency and product quality.

Although meta-transcriptomic analysis can provide valuable knowledge about the gene expression and potential functional activity, it might fail to establish direct associations between the microbiota and the environment, since the mRNA expression might not be directly associated with protein expression, and cell activity regulation occurs at the protein level. Therefore, the direct analysis of the proteins are essential as a complementary approach along with meta-transcriptomics to study the functional activity of the microbial community [ 103 ]. The meta-proteomic approach, which provides a large-scale study of the entire proteins expressed by a microbial community in an environmental sample, is useful for the identification and quantification of microbial activity at the post-translational level and could be the link between metagenomics studies and biological functions for understanding complex substrate-microbiome interactions [ 104 , 105 ]. Compared to other meta-omics approaches, meta-proteomic analysis is barely explored for studying food fermentation because of limitations, including the high cost, complexity of microbial samples, and the high similarity between many protein sequence reads [ 106 , 107 ]. Yang et al. provided a comprehensive review of the applications of proteomics in the study of fermented food and beverages [ 104 ].

5. Future Perspectives

The fermentation process has undergone significant improvement over the years. With technological advancement, it has become possible to manipulate the fermentation starter culture and the associated microbiome to standardise the product stability, improve sensory properties of the food, and ensure safety. Genome editing and microbiome engineering tools based on the CRISPR technology are evolving rapidly and are becoming highly efficient in improving microbial functionality. This modern technology is mostly applied in human cell research, and CRISPR/Cas9-based gene therapy has clinical potential. However, this technology still holds great promise for a wide range of applications, including in the food fermentation industry. The industry of fermented food and beverages may benefit significantly from the application of the revolutionary tools of CRISPR-based genome editing and microbiome manipulation, leading to improved control of the process and the properties of the end product by promoting the desired features linked to the sensory aspects, quality, shelf life, nutritional content, and safety of fermented food. This improvement would be coupled with increasing consumer acceptance and fewer governmental restrictions on the use of CRISPR/Cas9 gene editing in microbe manipulation, especially when no insertion of foreign DNA is involved. Scientific research is continuously proving the safety and precision of modern genomic manipulation tools, with no use of resistance markers and genetic stability of generated strains, thereby becoming accepted as a food-grade process.

However, it is important to highlight the worldwide popularity and acceptance of fermented food and its natural and artisanal nature. Therefore, spontaneous and traditional methods of preparation will continue to be a major part of the practice. Hence, the precise characterisation of the whole process and hygienic aspects are important to understand traditional fermentation and ensure the safety of consumption of fermented food. The available tools based on NGS technology and the rise of pioneering integrated multi-omics approaches have allowed deep understanding and high-resolution analysis of the fermentation process with many novel insight into the fermented-food microbiome and the role in the physicochemical and sensory properties of fermented food.

The advancement in tools available to study the fermentation process has revealed the inherited wisdom of ancient societies. The selection of suitable fermentation substrates to combine and set up the conditions of traditional fermentation led to the preparation of healthy products with improved sensory properties. The combined multi-omics approaches have provided cutting-edge discoveries in different microbiological research fields, while their application in studying fermentation is limited. Most studies use a single approach to study a particular aspect, especially because of the high cost of combining different omics approaches and the need for sophisticated bioinformatics and biostatistics skills for the analysis of such large datasets. A combined multi-omics approach would facilitate understanding the process, provide new systems-biology perspectives, and decipher the interaction among the fermentation microbiota, the substrate, and the environment. Overall, the near future will bring a greater range of applications of multi-omics in studying food fermentation, leading to detailed characterisation and efficient and sustainable production of fermented food and beverages.

Author Contributions

Conceptualisation, M.M. and Y.-S.S.; methodology, M.M. and G.H.; software, M.M. and G.H.; investigation, M.M. and I.P.; data curation, M.M., G.H. and Y.-S.S.; writing—original draft preparation, M.M. and G.H.; writing—review and editing, I.P. and Y.-S.S.; visualisation, M.M.; supervision, I.P. and Y.-S.S.; project administration, Y.-S.S.; funding acquisition, I.P. All authors have read and agreed to the published version of the manuscript.

This work was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) grant funded by the Ministry of Science, ICT and Future Planning (NRF-2018R1D1A1B07041112) and was supported by the BK21 Four Program of Pusan National University.

Conflicts of Interest

The authors declare no conflict of interest.

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T. Hasegawa expands R&D with new division, leadership

CERRITOS, Calif. — T. Hasegawa USA is integrating its food and beverage flavors divisions into a single research and development unit following its acquisition of Mission Flavors & Fragrances late last year.

The new business division combines food and beverage flavor development with applications expertise and will be led by an updated management team with several new director and senior flavor chemist positions.

Satoshi Koga, director of sweet technology at T. Hasegawa, will transition from his current role to lead the integration. He also will lead the evaluation of raw materials and processes to ensure consistent quality as the two companies continue merging. Ibrahima Faye, senior flavor chemist at T. Hasegawa, will succeed Mr. Koshi as director of sweet technology. Jeanene Martinez will lead the beverage applications team as director of applications.

“I’m excited to lead such a tremendous group of people who are instrumental in our research and development ensuring our customers success,” said Jim Yang, vice president of R&D at T. Hasegawa USA. “Our goal has always been to seek out team members that assist us in accelerating growth while directly impacting the rest of our team in a positive manner. There is nothing more important than to be able to find them from within our own company.”

Mr. Koga, Mr. Faye and Ms. Martinez were promoted alongside Maria Olson and Lauren Mayberry, who were named senior flavor chemists. Ms. Olson and Ms. Mayberry both joined the company via the Mission Flavors acquisition.

“We are thrilled to have this group enabling us to continued success in flavor creation,” Mr. Yang said. “This organizational change will fuel innovation, drive collaboration and accelerate our opportunities for growth at a pivotal time for T. Hasegawa.”

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Reimagining Design with Nature: ecological urbanism in Moscow

Reflective Essay
Published: 10 September 2019
Volume 1 , pages 233–247, ( 2019 )

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Brian Mark Evans ORCID: orcid.org/0000-0003-1420-1682 1

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The twenty-first century is the era when populations of cities will exceed rural communities for the first time in human history. The population growth of cities in many countries, including those in transition from planned to market economies, is putting considerable strain on ecological and natural resources. This paper examines four central issues: (a) the challenges and opportunities presented through working in jurisdictions where there are no official or established methods in place to guide regional, ecological and landscape planning and design; (b) the experience of the author’s practice—Gillespies LLP—in addressing these challenges using techniques and methods inspired by McHarg in Design with Nature in the Russian Federation in the first decade of the twenty-first century; (c) the augmentation of methods derived from Design with Nature in reference to innovations in technology since its publication and the contribution that the art of landscape painters can make to landscape analysis and interpretation; and (d) the application of this experience to the international competition and colloquium for the expansion of Moscow. The text concludes with a comment on how the application of this learning and methodological development to landscape and ecological planning and design was judged to be a central tenant of the winning design. Finally, a concluding section reflects on lessons learned and conclusions drawn.

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The landscape team from Gillespies Glasgow Studio (Steve Nelson, Graeme Pert, Joanne Walker, Rory Wilson and Chris Swan) led by the author and all our collaborators in the Capital Cities Planning Group.

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Evans, B.M. Reimagining Design with Nature: ecological urbanism in Moscow. Socio Ecol Pract Res 1 , 233–247 (2019). https://doi.org/10.1007/s42532-019-00031-5

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The Role of Environmental NGOs: Russian Challenges, American Lessons: Proceedings of a Workshop (2001)

Chapter: 14 problems of waste management in the moscow region, problems of waste management in the moscow region.

Department of Natural Resources of the Central Region of Russia

The scientific and technological revolution of the twentieth century has turned the world over, transformed it, and presented humankind with new knowledge and innovative technologies that previously seemed to be fantasies. Man, made in the Creator’s own image, has indeed become in many respects similar to the Creator. Primitive thinking and consumerism as to nature and natural resources seem to be in contrast to this background. Drastic deterioration of the environment has become the other side of the coin that gave the possibility, so pleasant for the average person, to buy practically everything that is needed.

A vivid example of man’s impact as “a geological force” (as Academician V. I. Vernadsky described contemporary mankind) is poisoning of the soil, surface and underground waters, and atmosphere with floods of waste that threaten to sweep over the Earth. Ecosystems of our planet are no longer capable of “digesting” ever-increasing volumes of waste and new synthetic chemicals alien to nature.

One of the most important principles in achieving sustainable development is to limit the appetite of public consumption. A logical corollary of this principle suggests that the notion “waste” or “refuse” should be excluded not only from professional terminology, but also from the minds of people, with “secondary material resources” as a substitute concept for them. In my presentation I would like to dwell on a number of aspects of waste disposal. It is an ecological, economic, and social problem for the Moscow megalopolis in present-day conditions.

PRESENT SITUATION WITH WASTE IN MOSCOW

Tens of thousand of enterprises and research organizations of practically all branches of the economy are amassed over the territory of 100,000 hectares: facilities of energy, chemistry and petrochemistry; metallurgical and machine-building works; and light industrial and food processing plants. Moscow is occupying one of the leading places in the Russian Federation for the level of industrial production. The city is the greatest traffic center and bears a heavy load in a broad spectrum of responsibilities as capital of the State. The burden of technogenesis on the environment of the city of Moscow and the Moscow region is very considerable, and it is caused by all those factors mentioned above. One of the most acute problems is the adverse effect of the huge volumes of industrial and consumer wastes. Industrial waste has a great variety of chemical components.

For the last ten years we witnessed mainly negative trends in industrial production in Moscow due to the economic crisis in the country. In Moscow the largest industrial works came practically to a standstill, and production of manufactured goods declined sharply. At the same time, a comparative analysis in 1998–99 of the indexes of goods and services output and of resource potential showed that the coefficient of the practical use of natural resources per unit of product, which had been by all means rather low in previous years, proceeded gradually to decrease further. At present we have only 25 percent of the industrial output that we had in 1990, but the volume of water intake remains at the same level. Fuel consumption has come down only by 18 percent, and the amassed production waste diminished by only 50 percent. These figures indicate the growing indexes of resource consumption and increases in wastes from industrial production.

Every year about 13 million tons of different kinds of waste are accumulated in Moscow: 42 percent from water preparation and sewage treatment, 25 percent from industry, 13 percent from the construction sector, and 20 percent from the municipal economy.

The main problem of waste management in Moscow city comes from the existing situation whereby a number of sites for recycling and disposal of certain types of industrial waste and facilities for storage of inert industrial and building wastes are situated outside the city in Moscow Region, which is subject to other laws of the Russian Federation. Management of inert industrial and building wastes, which make up the largest part of the general volume of wastes and of solid domestic wastes (SDW), simply means in everyday practice their disposal at 46 sites (polygons) in Moscow Region and at 200 disposal locations that are completely unsuitable from the ecological point of view.

The volume of recycled waste is less than 10–15 percent of the volume that is needed. Only 8 percent of solid domestic refuse is destroyed (by incineration). If we group industrial waste according to risk factor classes, refuse that is not

dangerous makes up 80 percent of the total volume, 4th class low-hazard wastes 14 percent, and 1st-3rd classes of dangerous wastes amount to 3.5 percent. The largest part of the waste is not dangerous—up to 32 percent. Construction refuse, iron and steel scrap, and non-ferrous metal scrap are 15 percent. Paper is 12 percent, and scrap lumber is 4 percent. Metal scrap under the 4th class of risk factor makes up 37 percent; wood, paper, and polymers more than 8 percent; and all-rubber scrap 15 percent. So, most refuse can be successfully recycled and brought back into manufacturing.

This is related to SDW too. The morphological composition of SDW in Moscow is characterized by a high proportion of utilizable waste: 37.6 percent in paper refuse, 35.2 percent in food waste, 10 percent in polymeric materials, 7 percent in glass scrap, and about 5 percent in iron, steel, and non-ferrous metal scrap. The paper portion in commercial wastes amounts to 70 percent of the SDW volume.

A number of programs initiated by the Government of Moscow are underway for the collection and utilization of refuse and for neutralization of industrial and domestic waste. A waste-recycling industry is being developed in the city of Moscow, mostly for manufacturing recycled products and goods. One of the most important ecological problems is the establishment in the region of ecologically safe facilities for the disposal of dangerous wastes of 1st and 2nd class risk factors.

Pre-planned industrial capacities for thermal neutralization of SDW will be able to take 30 percent of domestic waste and dangerous industrial waste. Construction of rubbish-burning works according to the old traditional approach is not worthwhile and should come to an end. Waste-handling stations have been under construction in the city for the last five years. In two years there will be six such stations which will make it possible to reduce the number of garbage trucks from 1,156 to 379 and to reduce the amount of atmospheric pollution they produce. In addition the switch to building stations with capacity of briquetting one ton of waste into a cubic meter will decrease the burden on waste disposal sites and prolong their life span by 4–5 fold. Trash hauling enterprises will also make profit because of lower transportation costs.

Putting into operation waste-segregation complexes (10–12 sites) would reduce volumes of refuse to disposal sites by 40 percent—that is 1,200,000 tons per year. The total volume of burned or recycled SDW would reach 2,770,000 tons a year. A total of 210,000 tons of waste per year would be buried. So, in the course of a five year period, full industrial recycling of SDW could be achieved in practice.

Collection of segregated waste is one of the important elements in effective disposal and utilization of SDW. It facilitates recycling of waste and return of secondary material into the manufacturing process. Future trends in segregation and collection of SDW will demand wide popularization and improvement of the ecological culture and everyday behavior of people.

In recent years the high increase in the number of cars in Moscow has brought about not only higher pollution of the atmosphere, but also an avalanche-like accumulation of refuse from vehicles. Besides littering residential and recreation areas, cars represent a source for toxic pollution of land and reservoirs. At the same time, automobile wastes are a good source for recycled products. In the short-term outlook, Moscow has to resolve the problem of collection and utilization of decommissioned vehicles and automobile wastes with particular emphasis on activities of the private sector. Setting up a system for collection and utilization of bulky domestic waste and electronic equipment refuse is also on the priority list.

In 1999 in Moscow the following volumes of secondary raw materials were produced or used in the city or were recycled: 300,000 tons of construction waste, 296,000 tons of metal scrap, 265 tons of car battery lead, 21,000 tons of glass, 62,500 tons of paper waste, 4,328 tons of oil-bearing waste, and 306 tons of refuse from galvanizing plants.

Such traditional secondary materials as metal scrap and paper waste are not recycled in Moscow but are shipped to other regions of Russia.

The worldwide practice of sorting and recycling industrial and domestic wastes demands the establishment of an industry for secondary recycling. Otherwise segregation of waste becomes ineffective.

There are restraining factors for the development of an effective system of assorted selection, segregation, and use of secondary raw resources, namely lack of sufficient manufacturing capacities and of suitable technologies for secondary recycling.

The problem of utilization of wastes is closely linked with the problem of modernization and sometimes even demands fundamental restructuring of industries. The practical use of equipment for less energy consumption and a smaller volume of wastes and a transition to the use of alternative raw materials are needed. Large enterprises—the main producers of dangerous wastes—are in a difficult financial situation now, which is an impediment for proceeding along these lines.

Private and medium-size enterprises are becoming gradually aware of the economic profitability in rational use of waste. For example, the firm Satory started as a transportation organization specialized in removal of scrap from demolished buildings and those undergoing reconstruction. It now benefits from recycling of waste, having developed an appropriate technology for the dismantling of buildings with segregation of building waste. So, as it has been already mentioned above, the first task for Moscow is to establish a basis for waste recycling.

HOW TO CHANGE THE SITUATION WITH WASTE

Transition to modern technologies in the utilization of wastes requires either sufficient investments or a considerable increase in repayment for waste on the part of the population. Obviously, these two approaches are not likely to be realized in the near future.

The recovery of one ton of SDW with the use of ecologically acceptable technology requires not less than $70–100.

Given the average per capita income in 1999 and the likely increase up to the year of 2005, in 2005 it will be possible to receive from a citizen not more than $14 per year. This means that the cost of technology should not exceed $40 per ton of recycled waste. Unfortunately, this requirement can fit only unsegregated waste disposal at the polygons (taking into account an increase in transportation costs by the year 2005).

Such being the case, it looks like there is only one acceptable solution for Russia to solve the problem of waste in an up-to-date manner: to introduce trade-in value on packaging and on some manufactured articles.

In recent years domestic waste includes more and more beverage containers. Plastic and glass bottles, aluminium cans, and packs like Tetrapak stockpiled at disposal sites will soon reach the same volumes as in western countries. In Canada, for example, this kind of waste amounts to one-third of all domestic waste.

A characteristic feature of this kind of waste is that the packaging for beverages is extremely durable and expensive. Manufactured from polyethylene terephthalate (PTA) and aluminum, it is sometimes more expensive than the beverage it contains.

What are the ways for solving the problem? Practically all of them are well-known, but most will not work in Russia in present conditions. The first problem relates to collection of segregated waste in the urban sector and in the services sector. A number of reasons make this system unrealistic, specifically in large cities. Sorting of waste at waste-briquetting sites and at polygons is possible. But if we take into account the present cost of secondary resources, this system turns out to be economically unprofitable and cannot be widely introduced.

The introduction of deposits on containers for beverages is at present the most acceptable option for Russia. This system turned out to be most effective in a number of countries that have much in common with Russia. In fact this option is not at all new for us. Surely, all people remember the price of beer or kefir bottles. A system of deposit for glass bottles was in operation in the USSR, and waste sites were free from hundreds of millions of glass bottles and jars. We simply need to reinstate this system at present in the new economic conditions according to new types and modes of packaging. Deposits could be introduced also on glass bottles and jars, PTA and other plastic bottles, aluminium cans, and Tetrapak packing.

Let us investigate several non-ecological aspects of this problem, because the ecological impact of secondary recycling of billions of bottles, cans, and packs is quite obvious.

Most of the population in Russia lives below the poverty line. When people buy bottles of vodka, beer, or soft drinks, they will have to pay a deposit value (10–20 kopeks for a bottle). The poorest people will carry the bottles to receiving points. A system of collection of packaging will function by itself. Only receiving points are needed. Millions of rubles that are collected will be redistributed among the poorest people for their benefit, and a social problem of the poor will be solved to a certain extent not by charity, but with normal economic means.

A second point is also well-known. In a market economy one of the most important problems is that of employment. What happens when the trade-in value is introduced?

Thousands of new jobs are created at receiving points and at enterprises that recycle glass, plastics, etc. And we don’t need a single penny from the state budget. More than that, these enterprises will pay taxes and consume products of other branches of industry, thus yielding a return to the budget, not to mention income tax from new jobs.

There is another aspect of the matter. Considerable funding is needed from budgets of local governments, including communal repayments for waste collection and disposal at polygons and incinerators. Reduction of expenses for utilization of waste can be significant support for housing and communal reform in general.

It is practically impossible to evaluate in general an ecological effect when thousands of tons of waste will cease to occupy plots of land near cities as long-term disposal sites. Operation costs of receiving points and transportation costs could be covered by funds obtained from manufacturers and from returned packaging. Besides, when a waste recycling industry develops and becomes profitable, recycling factories will be able to render partial support to receiving points.

Trade-in value can be introduced on all types of packaging except milk products and products for children. It could amount to 15 or 30 kopecks per container, depending on its size. If all plastic bottles with water and beer are sold with trade-in value only in Moscow, the total sum will reach 450 million rubles a year. If we include glass bottles, aluminum cans, and packets, the sum will be one billion rubles. This sum will be redistributed at receiving points among people with scanty means when they receive the money for used packaging and jobs at receiving points and at recycling factories.

The bottleneck of the problem now is the absence in Russia of high technology industries for waste recycling. It can be resolved rather easily. At the first stage, used packaging can be sold as raw material for enterprises, including those overseas. There is unrestricted demand for PTA and aluminum on the part

of foreign firms. When waste collection mechanisms are established, there will be limited investments in this branch of industry.

With regard to the inexhaustible source of free raw material, this recycling industry will become one of the most reliable from the point of view of recoupment of investments. The Government, regional authorities, the population, and of course ecologists should all be interested in having such a law.

The same should be done with sales of cars, tires, and car batteries. Prices of every tire or battery should be higher by 30–50 rubles. These sums of money should be returned back to a buyer or credited when he buys a new tire or a new battery. For sure, such being the case we will not find used batteries thrown about the city dumps. In this case the task is even simpler because there are already a number of facilities for the recycling of tires and batteries.

In fact, a law of trade-in value can change the situation with waste in Russia in a fundamental way. Russian legislation has already been prepared, and the concept of an ecological tax has been introduced in the new Internal Revenue Code. Now it needs to be competently introduced. The outlay for waste recycling has to become a type of ecological tax. To realize this task much work has to be done among the deputies and with the Government. Public ecological organizations, including international ones, should play a leading role.

ACTIVITY OF PUBLIC ORGANIZATIONS IN THE SPHERE OF WASTE MANAGEMENT IN THE MOSCOW REGION

We know examples of the ever increasing role of the general public in the solution of the problem of waste utilization, first of all in those countries that have well-developed democratic institutions. “Fight Against Waste” is one of the popular slogans of public organizations abroad. Public opinion has brought about measures of sanitary cleaning in cities, secured better work by municipal services, shut down hazardous industries, and restricted and prohibited incineration facilities. Nevertheless, the struggle against wastes in the economically developed countries, being a manifestation of an advanced attitude towards the environment, has in the long run brought about a paradoxical result. Transfer of hazardous industries to countries with lower environmental standards and inadequate public support—Russia, as an example—has made the world even more dangerous from the ecological point of view.

Russia has just embarked on the path of formation of environmental public movements by the establishment of nongovernmental organizations. Representatives of nongovernmental organizations from Russia took part in the international gathering in Bonn in March 2000 of nongovernmental organizations that are members of the International Persistent Organic Pollutants (POPs) Elimination Network. A declaration against incineration was adopted in

Bonn by nongovernmental organizations, which called for elaboration of effective alternative technologies for utilization of waste and safe technologies for elimination of existing stockpiles of POP.

Quite a number of environmental organizations are operating now in Moscow. First to be mentioned is the All-Russia Society for the Conservation of Nature, which was established in Soviet times. There are other nongovernmental organizations: Ecosoglasiye, Ecolain, Ecological Union, and the Russian branches of Green Cross and Greenpeace. All these organizations collect and popularize environmental information and organize protest actions against policies of the Government or local administrations on ecological matters. A new political party—Russia’s Movement of the Greens—is being formed.

Laws currently in force in the Russian Federation (“On Protection of the Environment,” “On State Ecological Examination by Experts,” “On Production and Consumption of Waste”) declare the right of the public to participate in environmental examination of projects that are to be implemented, including those on the establishment of facilities for elimination and disposition of waste. Public examinations can be organized by the initiative of citizens and public associations. For example, under the law of Moscow “On Protection of the Rights of Citizens while Implementing Decisions on Construction Projects in Moscow,” public hearings are organized by the city’s boards. Decisions taken by local authorities, at referenda and public meetings, may be the very reason for carrying out public examinations. Such examinations are conducted mainly by commissions, collectives, or ad hoc groups of experts. Members of public examination panels are responsible for the accuracy and validity of their expert evaluations in accordance with the legislation of the Russian Federation. A decision of a public environmental panel has an informative nature as a recommendation, but it becomes legally mandatory after its approval by the appropriate body of the State. Besides, the opinion of the public is taken into account when a project submitted for state environmental review has undergone public examinations and there are supporting materials.

Public environmental examination is supposed to draw the attention of state bodies to a definite site or facility and to disseminate well-grounded information about potential ecological risks. This important facet of public environmental organizations in Moscow and in Russia is very weak. To a large extent, it can be explained by an insufficient level of specific and general knowledge of ecology even on the part of the environmentalists themselves. Lack of knowledge on the part of ordinary citizens and public groups and inadequate information (for various reasons) produce alarm-motivated behavior by those who harm the organization of environmental activity in general and waste management in particular.

There are nevertheless positive examples of public participation in designing policies of local authorities in the waste management sphere.

Speaking about the Moscow region we can point to the very productive work of the Public Ecological Commission attached to the Council of Deputies in Pushchino, in Moscow Oblast.

The population of Pushchino is 21,000. The polygon for solid biological wastes (SBW) has practically exhausted its capacities. In 1996, in order to find a way out, the Administration of the town showed an interest in a proposal made by the Austrian firm FMW to support financially the construction of an electric power station in the vicinity of the town that would operate using both fuel briquettes and SBW of the town. The briquettes would be manufactured in Turkey and would contain 70 percent Austrian industrial waste with added oil sludge. It was also envisaged that during the construction period of the electric power station, 300,000 tons of briquettes would be shipped and stockpiled. The original positive decision was annulled due to an independent evaluation of the project organized by the Public Ecological Commission.

The general public of Puschino put forward a counter proposal before the Administration in order to reduce volumes of SBW disposal at the polygon and to prolong its operation—segregation of SBW (food waste, paper refuse, fabrics, metal, glass, used car batteries). As a result, a new scheme for sanitary measures in the town was worked out in 1998, which on the basis of segregation of waste provided for a considerable decrease in refuse flow to the polygon. Unfortunately, for lack of finances in the town budget, the scheme has not been introduced to the full extent. But in spite of severe shortages of special containers for segregated wastes, a network of receiving points for secondary materials was set up.

One of the pressing tasks for greater public activity is wide popularization of environmental knowledge on waste management, especially among the young generation. There is a very important role for public organizations to play in this domain when enlightenment and education are becoming a primary concern of nongovernmental organizations. Referring again to the example of the Public Ecological Commission in Pushchino, I have to underline that this organization is taking an active part in the enlightenment of the population through organizing exhibitions, placing publications in the press, and spurring school children into action to encourage cleaning of the town by means of environmental contests, seminars, and conferences. Children help the Commission organize mobile receiving points for secondary material. They even prepare announcements and post them around the town calling on the citizens to take valuable amounts of domestic wastes and car batteries to receiving points.

There are other examples of a growing influence of public organizations on the policy of administration in the sphere of waste management in the Moscow region. The Moscow Children’s Ecological Center has worked out the Program “You, He, She and I—All Together Make Moscow Clean,” which is being introduced with the support of the Moscow Government. In the framework of this program, children collect waste paper at schools, and they are taught how to

be careful about the environment and material resources. The storage facilities agreed to support the initiative. They buy waste paper at a special price for school children. Then, the schools spend the earned money for excursions, laboratory equipment, books, and plant greenery.

Another example of an enlightened activity is a project realized in 1999 by the firm Ecoconcord on producing video-clips for TV about the adverse effects of waste incineration and the illegality of unauthorized storage of waste.

The name Ecoconcord speaks for the main purpose of this organization—to achieve mutual understanding between the general public and governmental organizations, to encourage public involvement in decision-making, and to promote the formation of policy bodies that would not let public opinion be ignored.

Proceeding from the global task of integrating the activities of interested parties in lessening adverse waste pollution, public organizations have to cooperate with authorities and not stand against them. Cooperation and consensus between governmental and nongovernmental organizations in working out strategies and tactics in waste management should become a prerequisite in successful realization of state policy in this sphere in the Russian Federation.

An NRC committee was established to work with a Russian counterpart group in conducting a workshop in Moscow on the effectiveness of Russian environmental NGOs in environmental decision-making and prepared proceedings of this workshop, highlighting the successes and difficulties faced by NGOs in Russia and the United States.

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TH True Milk is known for its True Milk and other True products. © TH True Milk

TH Milk breaks ground on $630m milk processing plant in Russia

Vietnamese dairy company TH Milk has broken ground on a new milk processing facility in Russia as part of its continued investment in the country.

The plant, which will have a capacity of 1,500 tonnes of raw milk per day, will be built in Borovsk – around 90km southwest of Moscow and nearly 8,000km from Ho Chi Minh City.

TH Milk is investing more than $630 million in the plant – the largest single investment into Russian agriculture by a Vietnamese company.

It will source milk for the plant from five purpose-built farms in the Moscow and Kaluga regions of the country: the launch of the first farm for the production of raw milk in the Volokolamsky district of Moscow region took place in the first quarter of 2018.

The launch of the remaining four dairy farms in Moscow and Kaluga regions is scheduled for 2019 and 2020; TH Milk expects to launch a proprietary distribution network by 2020 and will reach full-scale production by 2025, allowing it to take a leading position in the country’s dairy sector.

The ground-breaking ceremony was attended by representatives of the Russian government, as well as the founder of TH Group, Thai Huong.

Kirill Dmitriev, CEO of the Russian Direct Investment Fund (RDIF), the Russian state’s investment vehicle, said: “RDIF’s partnership with leading Asian manufacturer TH Group will promote innovation in dairy production and have a positive impact on the development of the Russian dairy industry. Our joint investments have important social significance – the creation of new high-tech dairy plants will help to reduce the deficit of dairy products in the Russian market and provide consumers with high-quality domestic products.”

The project will utilise end-to-end milk processing technology, involving a complete production chain from growing fodder and milking herd to the production of raw milk and its processing into a wide range of final dairy products. The project provides innovative solutions for increasing the productivity of milking herd, as well as modern technological solutions, including automatic feeding and milking systems.

Vietnamese dairy companies are becoming increasingly important for Russia, as Western economies scale back their investments in the country and Moscow continues to place inhibitive restrictions on EU imports.

Earlier this year, the company announced it would invest $2.7 billion in its entire operation in the Russian market.

TH Milk founder Thai Huong said: “TH produces agricultural products of international quality standards. The cooperation of RDIF and TH Group will make a significant contribution to the restructuring of Russian Federation agriculture, giving Russian consumers the most benefit, which are good and healthy products purely from nature from their own land, and to give the Russians the right to be proud of their land.”

Tillamook president and CEO to step down

Arla Foods Ingredients showcases new concepts for nutrition-boosted cheese

Sargento expands cheese snacks range

Ben & Jerry’s launches limited-edition Marshmallow Sky ice cream

Danone to offload Russian assets in RUB 17.7bn deal – Financial Times

Proof Hard Ice Cream debuts new flavour at Costco

Nestlé Vietnam allocates $100m to expand coffee production facility

Global dairy companies join alliance to cut methane at COP28

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Published: 02 March 2022

The structure of microbial communities of activated sludge of large-scale wastewater treatment plants in the city of Moscow

Shahjahon Begmatov 1 ,
Alexander G. Dorofeev 2 ,
Vitaly V. Kadnikov 1 ,
Alexey V. Beletsky 1 ,
Nikolai V. Pimenov 2 ,
Nikolai V. Ravin 1 &
Andrey V. Mardanov 1

Scientific Reports volume 12 , Article number: 3458 ( 2022 ) Cite this article

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Microbial communities
Environmental biotechnology
Environmental microbiology
Microbial ecology
Water microbiology

Microbial communities in wastewater treatment plants (WWTPs) play a key role in water purification. Microbial communities of activated sludge (AS) vary extensively based on plant operating technology, influent characteristics and WWTP capacity. In this study we performed 16S rRNA gene profiling of AS at nine large-scale WWTPs responsible for the treatment of municipal sewage from the city of Moscow, Russia. Two plants employed conventional aerobic process, one plant—nitrification/denitrification technology, and six plants were operated with the University of Cape Town (UCT) anaerobic/anoxic/oxic process. Microbial communities were impacted by the technology and dominated by the Proteobacteria , Bacteroidota and Actinobacteriota . WWTPs employing the UCT process enabled efficient removal of not only organic matter, but also nitrogen and phosphorus, consistently with the high content of ammonia-oxidizing Nitrosomonas sp. and phosphate-accumulating bacteria. The latter group was represented by Candidatus Accumulibacter, Tetrasphaera sp. and denitrifiers. Co-occurrence network analysis provided information on key hub microorganisms in AS, which may be targeted for manipulating the AS stability and performance. Comparison of AS communities from WWTPs in Moscow and worldwide revealed that Moscow samples clustered together indicating that influent characteristics, related to social, cultural and environmental factors, could be more important than a plant operating technology.

The interplay between microbial communities and soil properties

Laurent Philippot, Claire Chenu, … Noah Fierer

High-throughput characterization of bacterial responses to complex mixtures of chemical pollutants

Thomas P. Smith, Tom Clegg, … Thomas Bell

Hidden diversity and potential ecological function of phosphorus acquisition genes in widespread terrestrial bacteriophages

Jie-Liang Liang, Shi-wei Feng, … Jin-tian Li

Introduction

The removal of numerous pollutants produced by agriculture, industry, and households is important for the protection of natural ecosystems and human health. Wastewater treatment plants (WWTPs) employ a series of mechanical and biological processes that convert contaminated water into a sufficiently clean state through a series of steps removing different types of organic and inorganic pollutants 1 , 2 . Typically, wastewater treatment in large facilities takes place in three stages. The first stage includes physical methods of water purification, the second stage—chemical and/or biological treatment in bioreactors with suspended or attached activated sludge (AS). The third stage is the final treatment of water and its disinfection.

At the second stage, a consortium of microorganisms of AS transforms pollutants into harmless products or into products less hazardous to the environment and humans than the original components 3 . AS is a taxonomically and metabolically diverse microbial community with complex trophic relationships between its members 4 . It is the largest managed artificial ecosystem, continuously functioning in many cases for decades. The composition of the microbial community, which is shaped by both operating conditions and influent characteristics 5 , 6 , determines the main biochemical processes of wastewater treatment, and its change, for example, the massive development of filamentous forms of bacteria, can lead to a decrease in the efficiency of treatment and the occurrence of emergency situations 7 .

In municipal wastewater treatment plants, microbial consortia of AS are often developed under similar conditions, since the content of the main components of wastewater is limited to a rather narrow range of concentrations (except in some extreme cases of highly diluted or concentrated wastewater), pH is usually 7–8, temperatures vary from 10 to 30 °C 8 . In addition, in modern technologies certain biologically determined rules are followed: maintaining the minimum aerobic age of sludge necessary for the development of nitrification (the retention time of solid matter), ensuring the optimal retention time of the sludge in the anaerobic zone for effective enhanced biological phosphorus removal, optimizing the ratio of biochemical oxygen demand, nitrogen and phosphorus, etc. 3 . Therefore, it can be expected that the composition of microbial communities of activated sludge will contain a common component, which was confirmed by the results of studies comparing the composition of AS communities of various WWTPs 6 , 9 , 10 . At the same time a high diversity and differences of microbial communities of AS were noted, which was associated with climatic factors and the specificity of certain treatment plants: the share of the industrial component in the total influent, the temperature regime, the peculiarity of the used technologies and the exploitation of plants 9 . It has been shown that there is a relationship between the diversity and composition of the microbial community and the performance of treatment facilities 2 , although the authors noted that the real effect is not the performance itself, but the variation of indicators such as chemical oxygen demand, the retention time of suspended matter etc.

Despite the extensive application of traditional and modern molecular methods for studying microbial consortia of AS, their ecophysiology, population dynamics and diversity are far from being comprehensively understood. Most microorganisms of activated sludge are not cultivated, and the role of many typical inhabitants is not clearly known 11 , 12 .

Recently, using a systematic worldwide sampling, a Global Water Microbiome Consortium (GWMC) analysed the 16S ribosomal RNA gene sequences from ~ 1,200 AS samples taken from 269 WWTPs 10 . This study revealed that although the global AS bacterial communities contain ~ 1 billion phylotypes, ASs has a small, global core bacterial community of 28 phylotypes that is strongly linked to WWTP performance 10 . This study showed that although the type of treatment process exerted significant effects on microbial community structures, it was overwhelmed by geographical separation, and the compositions of AS microbial communities were significantly different between any two continents 10 . Although the GWMC study included WWTPs from 23 countries on 6 continents, the distribution of samples was geographically biased and covered mostly North America, Western and Central Europe, Eastern Asia (mostly China), Australasia, and several cities in South America and South Africa 10 .

Another large-scale initiative, the MiDAS project, analyzed samples (mostly AS) from 740 WWTPs using different types of treatment technologies, and represents the largest global sampling of WWTPs to date 13 , 14 . The resulting full-length 16S rRNA gene reference database, MiDAS 4, represent a comprehensive catalogue of bacteria in wastewater treatment systems and taxonomy from the domain to species level 14 . Although this study targeted WWTPs located in 425 cities, 31 countries on 6 continents, like the GWMC project, MiDAS mostly covered the same geographic regions.

In order to expand the geographical coverage and our knowledge about microbiomes of AS in general, we analyzed the composition of microbial communities of AS at large-scale WWTPs in the city of Moscow (Russia). Although the Moscow WWTPs are among the largest in the world, they (as well as other WWTPs from Russia) were not studied in the framework of GWMC and MiDAS projects. There are only a few studies on the analysis of the composition of microbial communities of AS from Moscow WWTPs using modern high throughput molecular genetic methods. Kallistova et al . 15 using FISH method analyzed AS samples from four Moscow WWTPs and characterized the abundance of the major technologically important microbial groups (ammonium- and nitrite-oxidizing, phosphate-accumulating, foam-inducing, anammox bacteria, and methanogens) in the aeration tanks. Later Shchegolkova et al . 16 performed 16S rRNA gene profiling of AS communities in three WWTPs responsible for processing sewage with different origins: municipal wastewater, slaughterhouse wastewater, and refinery sewage. The taxonomic structures of AS microbiomes were found to become stable in time, and each WWTP demonstrated a distinct pattern 16 . Several studies were devoted to 16S rRNA profiling, metagenomics and FISH studies of nitritation/ anammox wastewater treatment bioreactors applied for the treatment of NH 4 -rich wastewater 17 , 18 , 19 , 20 .

In this study, we present the results of an analysis of the composition of AS microbial communities at nine large-scale WWTPs in Moscow employing three different technologies.

Characteristics of WWTPs in the city of Moscow

Wastewater treatment plants of JSC "Mosvodokanal" carry out the treatment of sewage in the city of Moscow. Household and industrial wastewater entering the city sewerage system undergoes a full purification process, including biological treatment with AS. The largest treatment facilities, the Lyuberetskiy and Kuryanovskiy WWTP complexes (hereafter referred to as LOS and KOS, respectively), each with a capacity of about 2 million m 3 per day, consist of several wastewater treatment units in which a number of modern technologies for biological wastewater treatment are implemented, including biological nutrient removal (carbon, nitrogen, phosphorus) 21 , 22 , 23 , 24 . These treatment units (hereafter referred to as WWTPs) at each WWTP complex are fed by the same inflow water but otherwise are independent installations between which there is no transfer and mixing of AS.

After the primary treatment, the wastewater is subjected to purification in bioreactors with suspended activated sludge. The characteristics of the wastewater entering bioreactors at the two WWTP complexes are somehow different. The waters at the Kuryanovskiy WWTP complex contain approximately 1.5 times lower concentrations of organic matter and phosphorus compared to the Lyuberetskiy WWTPs, while the ammonium content do not differ significantly (Table 1 ).

All bioreactors are continuous: the inflowing wastewater and the returned AS are continuously fed to the inlet (or to a certain point) of the bioreactor, where they are mixed. Then the mixed liquor passes the bioreactor and enters the clarifier where the sludge is separated from the outflowing purified water by sedimentation. Chemical coagulants are not used. Three main technologies are used in the investigated WWTPs (Fig. 1 ).

Schemes of three types of bioreactors showing the different compartments and flow directions. Anaerobic, anoxic and oxic zones are colored in green, violet and blue, respectively.

In the simplest process, implemented at WWTPs 1 and 7, the oxidation of organic matter by heterotrophic aerobic microorganisms and the oxidation of ammonium by nitrifiers with the formation of nitrate are carried out in plug-flow bioreactors under aerobic conditions. The sludge mixture moves continuously along an elongated corridor from the point of mixing of the return AS with incoming wastewater to the clarifier. Wastewater is supplied either only at the beginning of the bioreactor (Conventional Activated Sludge process, CAS) (WWTP7), or along the entire aeration tank corridor (Step Feed Process, SF) (WWTP1). The entire volume of the bioreactor is aerated by blowing air.

The nitrification/ denitrification technology (N-DN) is realized at WWTP2. Wastewater is fed into the plug-flow bioreactor at the beginning and in the middle of the bioreactor and passes through zones without aeration, each followed by an aerobic zone. In these two anoxic zones, denitrification occurs due to the organic matter contained in the wastewater and nitrates contained in the return AS mixture (in the first anoxic zone) or arriving from the first aerobic zone (in the second anoxic zone). In two aerated zones organics and ammonium are oxidized.

Six other WWTPs are operated by anaerobic/anoxic/oxic process, also known as the University of Cape Town (UCT) technology. At the first stage, the sludge mixture enters the anaerobic zone, where phosphate-accumulating microorganisms (PAO) consume easily degradable organic matter, then to the anoxic zone, where denitrification and accumulation of phosphates by denitrifying PAO occur, and finally to the aerobic zone, where organic matter and ammonium are oxidized. Recycling from the aerobic zone ensures the inflow of nitrates into the anoxic zone, and the recycle of the AS mixture from the end of anoxic zone into the anaerobic zone minimizes the ingress of nitrates. Plug-flow bioreactors are used at WWTPs 4, 8 and 9, and carousel bioreactors at WWTPs 3, 5 and 6 (Supplementary Fig. S1 ).

The bulk dissolved oxygen concentration in aerobic zones of all bioreactors was 2–3 mg/L, the solid retention time was 15–25 days, and the hydraulic retention time was 8–12 h. The temperature in the time of AS sampling was 18–20 °C, and did not fall below 16 °C even in winter. The concentrations of AS were similar (2–4 g/L) in all WWTPs, and volatile suspended solids accounted for 60–65%.

Sampling and chemical analysis

The AS samples were obtained from nine wastewater treatment facilities at at Luberetsky and Kuryanovsky WWTP complexes in the Moscow region between March and May 2021. Samples of return activated sludge at 9 WWTPs (each in three replicas, 27 samples in total) were taken in BD Falcon tubes and immediately transferred to the laboratory at + 4 °C.

Samples of inflow and outflow water were collected between December 2020 and May 2021 and kindly provided by “Mosvodokanal” JSC. Chemical analysis was carried out according to standard methods. Water quality values (biochemical oxygen demand (BOD), chemical oxygen demand (COD), ammonium nitrogen (NH 4 -N) and phosphorus (P-PO 4 ) of influent and effluent, as well as nitrate nitrogen (NO 3 -N) in the effluent were measured using environmental standard methods twice a week for 6 months. Monthly average values are shown in Table 1 .

DNA isolation, amplification and sequencing of the 16S rRNA gene fragments

Total genomic DNA from each AS sample was extracted using a Power Soil DNA isolation kit (MO BIO Laboratories, Inc., Carlsbad, CA, USA) and stored at − 20 °C. PCR amplification of 16S rRNA gene fragments comprising the V3–V4 variable regions was carried out using the universal prokaryotic primers PRK 341F (5′-CCTAYG GGDBGCWSCAG) and PRK 806R (5′-GGA CTA CNVGGG THTCTAAT) 25 . The PCR fragments were bar-coded using the Nextera XT Index Kit v.2 (Illumina, San Diego, CA, USA) and purified using Agencourt AMPure beads (Beckman Coulter, Brea, CA, USA). The concentrations of PCR products were determined using the Qubit dsDNA HS Assay Kit (Invitrogen, Carlsbad, CA, USA). All PCR fragments were then mixed and sequenced on Illumina MiSeq (2 × 300 nt from both ends). Pairwise overlapping reads were merged using FLASH v.1.2.11 26 . The final dataset consisted of 2,173,862 16S rRNA gene reads (Supplementary Table S1 ).

Bioinformatics analysis of microbial community composition and diversity

All sequences were clustered into operational taxonomic units (OTUs) at 97% identity using the USEARCH v. 11 program 27 . Low quality reads, chimeric sequences, and singletons were removed by the USEARCH algorithm. To calculate OTU abundances, all reads obtained for a given sample (including singletons and low-quality reads) were mapped to OTU sequences at a 97% global identity threshold by USEARCH. The taxonomic assignment of OTUs was performed by searching against the SILVA v.138 rRNA sequence database using the VSEARCH v. 2.14.1 algorithm 28 . The recently developed MiDAS 4 14 , a full-length 16S rRNA gene reference database for wastewater treatment systems, was used to taxonomically identify OTUs up to the species level in the same way.

The diversity indices at a 97% OTU cut-off level were calculated using Usearch v.11 27 . To avoid sequencing depth bias, the number of reads generated for each sample were randomly sub-sampled to the size of the smallest dataset (94,942 reads) using the «single_rarefaction.py» script of QIIME 29 .

Calculation of Jaccard and weighted Unifrac distance metrics and trees was performed applying “beta_div” command in USEARCH. For the UniFrac analyses, a tree for the OTUs based on the sequence identity was constructed in USEARCH using “cluster_agg” command.

Integration of data from this study with data from GWMC

All 16S rRNA gene sequences assigned to OTUs obtained in the present work were mapped to OTU sequences from GWMC ( http://gwmc.ou.edu/ ). About 95% of sequences obtained in our experiments were mapped to GWMC OTUs at a 97% global identity threshold. OTU tables of both sets of samples were merged using command of “otutab_merge” of USEARCH. The obtained combined OTU table served as the input data for constructing a neighbor-joining tree generated from the Bray–Curtis dissimilarity matrix, which was calculated using the “beta_div” command of the Usearch program. The obtained tree was annotated and visualized in R package using ggtree method 30 , 31 .

Network analysis

Co-occurrence networks were inferred based on a Spearman correlation matrix 32 and constructed using only significant correlation 33 . The cutoff for correlation coefficients was determined to be 0.6 and the cutoff for adjusted p -values was 0.001 34 . Only OTUs, the relative abundance of which was at least 0.5% in at least one sample, were included in the analysis. Visualization of co-occurrence network was performed using Cytoscape v.3.8.2 platform 35 , 36 .

Data availability

The raw data generated from 16S rRNA gene sequencing have been deposited in the NCBI Sequence Read Archive (SRA) and are available via the BioProject PRJNA764866.

Results and discussion

Performance characteristics of wwtps.

Three main technologies are used in the investigated WWTPs. The simplest aeration process is implemented at WWTPs 1 and 7, where wastewater is supplied either at the beginning of the bioreactor corridor (CAS process, WWTP7), or along the entire aeration tank (SF process, WWTP1). The WWTP1 removed more than 98% of organics (according to the BOD data) and about 90% of ammonium, while the purification efficiency of WWTP7 was lower (95% removal of organics and 70% of ammonium) (Table 1 ). Interestingly, although these units were not designed to remove phosphorus, the WWTP7 removed more than 70% of phosphorus, while in WWTP1 its removal was inefficient. The nitrification/ denitrification technology, realized at WWTP2, enabled removal of more than 98% of organics and more than 99% of ammonium, while phosphorous was not removed (Table 1 ). Six WWTPs operated by the UCT technology enabled efficient purification of the wastewater from both organics (> 98%), ammonium (> 99%) and phosphorous (> 90%) (Table 1 ). The concentrations of N-NO 3 in the effluent were much lower than N-NH 4 in the influent indicating efficient denitrification in all WWTPs (Table 1 ).

Diversity of microbial communities of AS

Between 23,167 and 84,634 16S rRNA gene sequences were obtained for 27 analysed AS samples (9 WWTPs, 3 replicas) and clustered into 14,690 OTUs at the level of 97% identity. Neighbour-joining trees based of the UniFrac analysis (Fig. 2 ) and Jaccard similarity (Supplementary Fig. S1 ) of OTU datasets revealed that replicate samples formed distinct branches for all WWTPs except for WWTP8 and WWTP9 which use identical bioreactors and treatment technologies. Therefore, for subsequent analysis, for each of the 9 WWTPs, three replicates were combined into one dataset. Both UniFrac and Jaccard trees revealed clustering of samples according to the technology used (Fig. 2 and Supplementary Fig. S2 ).

Neighbor joining tree illustrating weighted UniFrac distances between microbial communities of AS samples from 9 WWTPs (three replications). Sample IDs are shown in brackets after the WWTP number.

The number of species-level OTUs present in AS at individual WWTPs ranged between 3860 and 4868, these values are typical for large-scale wastewater treatment plants 10 . Overall, the diversity of microbial communities did not significantly vary between WWTPs employing different technologies, while the evenness of microbial communities in plants operated by the UCT process was slightly lower than in the others (Supplementary Table S2 ).

Microbial community patterns at the phylum level

Taxonomic assignment of OTUs revealed the presence of 53 phylum-level lineages of Bacteria and Archaea, recognized in the Genome Taxonomy Database (GTDB) 37 . However, top 11 phyla comprising on average more than 1% of all the 16S rRNA gene sequences together accounted for more than 90% of the community (Fig. 3 and Supplementary Table S3 ).

Bacterial and archaeal community composition in AS samples according to the results of 16S rRNA gene sequencing. The composition is displayed at the phylum level. Average values for three replicas are shown for each WWTP.

Archaea represented less than 2% of sequences in all samples and were assigned to the phyla Nanoarchaeota, Halobacterota, Euryarchaeota and Thermoplasmatota ; each was detected in all samples. Besides members of the phylum Nanoarchaeota , known to comprise partner-dependent parasites or symbionts with small genome size and limited metabolic capacities 38 , most of other Archaea represented known methanogenic lineages of the families Methanobacteriaceae and Methanosaetaceae .

Bacterial communities were dominated by the Proteobacteria (on average 27.8% of 16S rRNA gene reads), mostly of classes gamma (23.5%) and alpha (4.3%). Other major groups were the Bacteroidota (15.7%), Actinobacteriota (12.5%), Chloroflexi (6.6%), Myxococcota (5.9%), Firmicutes (5.6%), Patescibacteria (5.5%), Verrucomicrobiota (4.5%), Bdellovibrionota (3.9%), Nitrospirota (2.7%), and Planctomycetota (1.3%). The relative abundances of these lineages in different samples differ by no more than several times, with the exception of Nitrospirota , the share of which is minimal (0.07%) in WWTP3 and maximal (7.1%) in WWTP2. All other bacterial phyla accounted for less than 2% of 16S rRNA gene reads in all samples except for the Campylobacterota representing about 3.5% of the community in WWTP1.

The main microbial drivers of wastewaters treatment

The biological wastewater treatment includes several microbial-driven processes, such as mineralization of organics by heterotrophs, oxidation of ammonia to nitrite and finally to nitrate, denitrificartion with the production of N 2 gas, enhanced biological phosphorus removal etc. In this section we analyze the presence of the microbial groups which could be involved in these processes. The activities of microbial processes depend on the absolute concentrations of microorganisms, but given that the concentration of AS in all bioreactors was similar, we can consider the efficiency of processes in relation to the relative abundance of the corresponding functional group in the community. Data on the shares of the discussed groups of microorganisms are shown in Table 2 .

Oxidation of ammonia and denitrification

Oxidation of ammonia to nitrate via nitrite is usually accomplished by two groups of microorganisms, although complete oxidation of ammonia to nitrate by Nitrospira sp. (comammox process) has been also reported 39 . Among known ammonia oxidizers (AOM) only members of the family Nitrosomonadaceae , mostly of the genus Nitrosomonas , were detected. The relative abundance of Nitrosomonadaceae clearly correlated with efficiency of ammonia removal: it ranged from 0.8% to 2.9% in all WWTPs enabling good removal of ammonia (> 99%) and was below 0.4% in WWTPs 1 and 7 where ammonia removal was less efficient.

Two genera of nitrite-oxidizing bacteria (NOB) were identified, Nitrospira and Candidatus Nitrotoga. The relative abundance of Nitrospirae sp. was the highest (7.06%) in WWTP2, which uses nitrification–denitrification process and much lower in WWTPs 1 and 7 (0.78% and 0.08%). WWTPs using the UCT process in terms of Nitrospirae content were clearly divided into two groups. AS samples of WWTPs 6, 8 and 9 harbored between 3.9% and 6.5% of Nitrospirae sp., while its relative abundance was less than 0.5% in WWTPs 3, 4 and 5. However, microbial communities of these three bioreactors contained 0.26 to 0.46% of Ca. Nitrotoga, nearly absent in other WWTPs. This recently described Ca . Nitrotoga species can be functionally and sometimes dominant important nitrite oxidizers in WWTPs due to their relatively higher resistance to free nitrous acid and free ammonia than other NOB and the presence of complete pathways for hydrogen and sulfite oxidation, suggesting that alternative energy metabolisms enable these bacteria to survive nitrite depletion 40 , 41 , 42 .

Regardless of the used purification technology and the efficiency of ammonia removal, the concentration of N-NO 3 in the effluent was in the range from 7 to 10.5 mg/L, which corresponds to 15–20% of N–NH 4 in the influent and indicates effective denitrification and removal of nitrogen in the gaseous form. The presence of numerous heterotrophic denitrifying bacterial is consistent with this observation.

Biological phosphorus removal

The next main issue of wastewater treatment is the removal of phosphorus. This process is carried out by microorganisms of the group of phosphate-accumulating organisms (PAO) capable of intracellular accumulation of polyphosphates under cyclic growth conditions, with alternated presence and absence of electron acceptors 43 , 44 , 45 , 46 .

Typical PAO, most often found in WWTPs, are members of the candidate species Candidatus Accumulibacter phosphatis (family Rhodocyclaceae , Gammaproteobacteria ) 47 , 48 , 49 , 50 . During the anaerobic phase, Ca . Accumulibacter phosphatis takes up volatile fatty acids present in the wastewater and stores the carbon from these substrates intracellularly as polyhydroxyalkanoates 51 . At the same time, intracellular polyphosphate is degraded to form ATP, releasing phosphate into the medium. During the aerobic phase, stored polyhydroxyalkanoates are used for energy production while phosphate is taken up from the medium and accumulated as polyphosphate. Ca . Accumulibacter phosphatis were found in all AS samples. Its relative abundance was the lowest (0.12%) in WWTP2 employing the nitrification/ denitrification process and poorly removing phosphorus. In other plants Ca . Accumulibacter phosphatis accounted for 0.43 to 1.05% of the communities and its share does not correlate with the phosphorus removal efficiency or the type of treatment process.

Several other bacteria, despite the difference in their metabolism from Ca. Accumulibacter phosphatis, are considered as likely PAO 46 , including members of Proteobacteria ( Dechloromonas 52 , 53 , Zooglea 54 , 55 , Thauera 56 , Thiothrix 57 , Ca . Accumulimonas 58 , Malikia 59 ), Actinobacteria ( Tetrasphaera 60 , 61 , Microlunatus 62 , Candidatus Microthrix 63 ), Gemmatimonadetes ( Gemmatimonas 64 ) and Melainabacteria ( Candidatus Obscuribacter phosphatis 65 ). Among these bacteria, members of the genera Tetrasphaera, Zoogloea, Thiothrix, Dechloromonas, Ca. Obscuribacter, Gemmatimonas , and Ca. Microthrix were detected.

The most abundant potential PAO, Ca. Microthrix sp., (average share 6.8%) was presented in all WWTPs and its share was higher in WWTPs employing the UCT process than in other types of bioreactors. Earlier it has been shown that Candidatus Microthrix parvicella contained of large polyphosphate granules and this microbial group might have been responsible for phosphorus removal during the sludge bulking period when Ca . Accumulibacter phosphatis was excluded from the system 63 . Dechloromonas sp., denitrifying bacteria capable of acetate uptake and polyphosphate storage, accounted for 1.5% to 5.8% of AS microbiomes in bioreactors operating under aerobic and UCT processes, while in WWTP2 it accounted for only 0.6%. Members of the genera Zoogloea and Thiothrix , aerobic bacteria capable of denitrification, often occur in ASs 57 , 66 , 67 . Both genera were more abundant in WWTPs employing aerobic process. Ca. Obscuribacter was most frequent in the WWTP6 (about 1.9%) and accounted for less than 1% in other samples. Members of the genus Tetrasphaera , known to be capable of aerobic polyphosphate accumulation under condition of assimilating glucose and/or amino acids anaerobically in advance 68 were found in all WWTPs. Their relative abundance was minimal in WWTPs 8 and 9 (about 0.2%) and reached 1.35% in WWTP1. Although nitrate-reducing bacteria of the genus Thauera are primary known for their ability to perform anaerobic degradation of aromatic and other refractory compounds 69 , recently Thauera sp. strain SND5 was found to be a phosphate-accumulating organism 56 . The relative abundance of Thauera sp. ranged from 0.2 to 2.2% and was maximal in WWTP8.

Glycogen accumulating organisms (GAOs) can accumulate glycogen and polyhydroxyalkanoates inside cells, but do not have ability to intracellular accumulation of polyphosphates. They are commonly found together with PAO in EBPR bioreactors where their predominance may lead to EBPR failure due to the competition for carbon source with PAO 70 . Known GAO belongs to the Gammaproteobacteria ( Candidatus Competibacter phosphatis) and Alphaproteobacteria ( Defluvicoccus sp . ) 71 , 72 . Ca. Competibacter accounted for up to 8.5% of the communities in UCT WWTPs, but were less numerous (< 1%) in WWTPs 2 and 7. The relative abundance of Defluvicoccus sp. was less than 0.1% in all samples. Thus, in WWTPs using the UCT process, a high relative abundance of GAO did not adversely affected the efficiency of phosphorus removal.

Microbial community composition and efficiency of wastewater purification

All five WWTPs using the UCT process ensured effective removal of not only organic matter, but also nitrogen and phosphorus, which is consistent with the high content of nitrifying and phosphate accumulating bacteria.

WWTP2, using the nitrification–denitrification process, effectively removed nitrogen, and the relative abundance of nitrifiers in this AS community was the highest. However, there was almost no phosphorus removal, and the share of PAO was the lowest. The low efficiency of phosphorus removal and the low abundance of PAO in WWTP2 are associated with the absence of anaerobic zones in bioreactors, as well as the consumption of easily degradable organic matter by denitrifying microorganisms.

Contrary to expectations, the two WWTPs, using a simple aeration process, provided the poorest removal of both organics and ammonia, while the nitrate content in the effluent was approximately the same as in other installations. However, the total nitrogen content in the form of nitrate and ammonium in the effluent was much lower than in the influent, indicating that nitrogen was removed during denitrification. Probably, under conditions of insufficient aeration, formation of local anoxic zones within microbial flocs facilitates the denitrification performed by heterotrophic bacteria 73 , while part of the ammonia and organic matter remain non-oxidized and enters the effluent water.

Interestingly, relatively efficient phosphorus removal was observed in WWTP7 but not in WWTP1. These two bioreactors differ in that in WWTP7 the organic-rich influent is fed to the beginning of the bioreactor, while in WWTP1 it is distributed along the entire length. It is likely that in WWTP7 local organic-rich anaerobic zones are formed within AS flocks near the point of influent supply, while in WWTP1 the concentration of organic substances is everywhere lower. In such organic-rich zones denitrifying PAO of the genera Zoogloea and Thiothrix can proliferate and their shares were maximal in WWTP7.

Other abundant community members

Analysis of the compositions of microbial communities revealed 16 genus-level lineages, the average share of which across nine WWTPs exceeds 1% (Supplementary Table S4 ). In total, these 16 genera accounted for 28 to 51% (on average about 36%) of analyzed microbiomes. The most abundant genus, Ca . Microthrix, was the most numerous in AS samples of WWTPs 4, 6, 8 and 9 (11—13%) using the UCT process, while in other samples its relative abundance was much lower (1.2—3.4%). Besides its potential role in phosphorus removal, Ca . Microthrix is known as a slowly growing filamentous bacterium causing bulking and foaming in activated sludge systems thus seriously affecting the stable operation of WWTPs 74 . The growth of cultured species of this genus, M. parvicella , is susceptible to both environmental and operational parameters, such as temperature, oxygen concentration, type of substrate, electron acceptor, organic loading, and sludge retention time (reviewed in 75 ). Probably, enrichment of Ca . Microthrix in the AS of the UCT WWTPs was associated with low oxygen concentrations in the anaerobic and anoxic zones of these bioreactors, favorable for the growth of these bacteria 7 . The dominant Ca . Microthrix phylotype was identified as Ca Microthrix subdominans, a species typically present along with Ca . M. parvicella, although usually in lower abundances 76 .

Besides Ca . Microthrix and above mentioned Ca . Competibacter, Dechloromonas, Nitrospira, Nitrosomonas and Thauera, the list of most abundant genera includes 3 cultured (Trichococcus , Haliangium , and Chitinivorax ) and 7 uncultured lineages, defined in the Silva database as OM27_clade ( Bdellovibrionota, Bdellovibrionaceae ), Candidatus Nomurabacteria ( Patescibacteria ), AKYH767 ( Bacteroidota, Sphingobacteriales ), a member of the family Moraxellaceae ( Gammaproteobacteria ) defined as “Agitococcus lubricus group’, mle1-27 ( Myxococcota ), env.OPS_17 ( Bacteroidota, Sphingobacteriales ), and OLB12 ( Bacteroidota, Microscillaceae ).

Members of the genera Trichococcus, Haliangium , and Chitinivorax are often found in AS of wastewater treatment facilities. Trichococcus sp. are filamentous bacteria that can degrade a wide range of carbohydrates 77 , while Chitinivorax sp. are devoted to the hydrolysis of chitin 78 . Similarly to Dechloromonas sp., members of the Haliangium are active denitrifiers in the AS communities 79 .

All uncultured genus-level lineages were previously detected in bioreactors treating wastewater 80 , 81 , 82 , 83 , but their functional roles remain elusive. The most abundant uncultured OM27_clade, with an average share of about 3%, was hypothesized to be predatory bacteria 84 . Members of the candidate genus OLB12 are aerobic heterotrophs abundant in the anammox granules collected from partial-nitritation anammox reactor 85 . The only cultured member of “Agitococcus lubricus group’, A. lubricus , is a lipolytic aerobic heterotroph found in freshwater bodies 86 . The relative abundance of this lineage reached 4% in WWTP2, while it was below 0.2% in all AS samples currently described in the MiDAS database 14 .

Various functional characteristics of microorganisms are closely related to the differentiation of ecological niches, and network analysis allows one to deduce patterns of interactions that develop within the WWTP ecosystem. Network analysis indicated possible cooperation (co-occurrence) and mutual exclusion among diverse microorganisms in WWTPs (Fig. 4 ).

Network of co-occurring abundant microbial OTUs of AS based on correlation analysis. A connection stands for a strong (Spearman’s rho > 0.6) and significant (adjusted p value < 0.001) correlation. OTUs are colored according to phylum. Co-presence and mutual exclusion of OTUs are shown in white and red lines, respectively.

Of 104 OTUs, with abundances more than 0.5% in at least one of the AS samples, 83 had statistically significant connections with other OTUs the maximum number of which reached 18 (Supplementary Table S5 ). OTUs with a high number of connections (“degree”) are considered as hub members, supposed to be key players in sustaining the overall community 87 . The largest number of connections (18) was observed for OTU assigned to the genus Haliangium , whose members, together with Dechloromonas sp., are active denitrifiers in WWTPs communities 79 . Notably, bacteria participating in ammonia and phosphorous removal were among the hub members, namely, Nitrosomonas , Nitrospira , Ca . Competibacter, Dechloromonas, and Thauera . Positive connection was detected between denitrifiers ( Thauera Otu26 and Haliangium Otu95) and bacteria involved in the oxidation of ammonia ( Nitrosomonas Otu18 and Nitrospira Otu2). This link is consistent with the fact that the source of nitrate is mainly microbial oxidation of ammonium, rather than influent sewage water. Interestingly, we did not detected mutual exclusion between Ca . Accumulibacter and Ca . Competibacter.

The nest largest number of connections was detected for Otu105, assigned to the candidate genus Sumerlaea (candidate phylum Sumerlaeota , previously known as BRC1). Members of this genus were predicted to be metabolically versatile bacteria capable to utilize various carbohydrates, including chitin, performing fermentation as well as complete oxidation of organic substrates through aerobic and anaerobic (with nitrate and sulfur compounds) respiration 88 . Co-presence of Sumerlaea and denitrifiers ( Thauera and Haliangium ) suggest that Sumerlaea sp. could be involved in degradation of organic substrates linked to dissimilatory nitrate reduction. Interestingly, no connection to other community members was found for the most abundant species, Ca . Microthrix (Otu1). Ca . Microthrix has been considered a specialized consumer of long-chain fatty acids that have been confirmed to be the key carbon and energy sources for its growth both under aerobic, anoxic and anaerobic conditions 75 , 89 . Probably due to such metabolic specialization Ca . Microthrix has limited interactions with other community members.

Microbial communities of Moscow WWTPs on a global scale

Variations in community composition are key points for understanding community functioning. To understand how the composition of microbial communities of AS varied across different geographic locations, we compared taxonomic structures of our microbiomes with that reported in a worldwide study performed by a Global Water Microbiome Consortium 10 . That study showed that although the taxonomic community structures observed on different continents were not clearly separated at the OTU level in two dimensional ordinations, it was significantly different between any two continents 10 , and although the type of treatment process exerted significant effects on microbial community structures, it was overwhelmed by continental geographical separation 10 .

Despite its high diversity, AS microbiome has a small global core bacterial community (28 OTUs) that is strongly linked to WWTP performance and accounted for an average 12.4% of the 16S rRNA gene sequences in AS samples 10 . 26 of these OTUs were present in Moscow WWTPs and accumulated from 11.3 to 23.5% of 16S rRNA gene reads (Supplementary Table S6 ).

Clustering based on the Bray–Curtis dissimilarity showed that all samples from Moscow WWTPs clustered together and formed a distinct branch not embedded into European or any other large groups, but forming a sister lineage to a subsets of Asian, South American and North American clusters (Fig. 5 ). These data indicate that influent characteristics, related to cultural, social and environmental factors in each region, could be more important than a plant operating conditions. Similar observations have been previously reported for the microbial communities of AS from WWTPs in Korea and Vietnam 2 . For example, due to the relatively low cost of water for household consumption, wastewater in Moscow has a relatively low content of organic matter compared to the values typical for most large cities (COD of 600–900 mg/L). It should be noted that the primers used in our work for the 16S rRNA gene profiling differed from those used by the GWMC consortium (515F/806R), which may affect the observed shares of particular microbial taxa. However, both primer pairs (515F/806R and 341F/806R) enabled good coverage of overall microbial diversity and provided similar estimates of the relative abundancies of most species in various environments 90 , 91 .

Neighbor joining tree illustrating clustering based on the Bray–Curtis dissimilarity between microbial communities of AS samples from 9 WWTPs obtained in this study and ones reported by a Global Water Microbiome Consortium ( http://gwmc.ou.edu/ ). Samples are colored according to geographic location.

Conclusions

This study provides the data on the composition of microbial communities of AS from large-scale WWTPs of the Moscow city and the evidences of the impact of purification technology. Although the taxonomic structure of communities at high ranks was similar and the most numerous groups were found in all WWTPs, UniFrac and Jaccard trees revealed clustering of samples according to the employed technology. WWTPs employing the UCT process enabled efficient removal of organics, nitrogen and phosphorus, while WWTPs operated under simple aeration or nitrification–denitrification process removed these components less efficiently. Co-occurrence network analysis provided information on key hub microorganisms in AS, which may be targeted for manipulating the AS stability and performance. Comparison of AS communities from Moscow WWTPs with ones analyzed worldwide by GWMC revealed that all Moscow samples clustered together, indicating that influent characteristics, related to cultural, social and environmental factors in each region, could be more important than a plant operating conditions.

Ahkola, H. et al. A preliminary study on the ecotoxic potency of wastewater treatment plant sludge combining passive sampling and bioassays. Sci. Total. Environ. 758 , 143700 (2021).

ADS CAS PubMed Google Scholar

Kim, Y. K. et al. The capacity of wastewater treatment plants drives bacterial community structure and its assembly. Sci. Rep. 9 , 14809 (2019).

ADS PubMed PubMed Central Google Scholar

Tchobanoglous, G., Burton, F.L. & Stensel, H.D. Wastewater Engineering: Treatment and Reuse . 1819 (McGraw-Hill Education, 2003).

Wagner, M. & Loy, A. Bacterial community composition and function in sewage treatment systems. Curr. Opin. Biotechnol. 3 , 218–227 (2002).

Google Scholar

Ibarbalz, F. M., Figuerola, E. L. & Erijman, L. Industrial activated sludge exhibit unique bacterial community composition at high taxonomic ranks. Water Res. 11 , 3854–3864 (2013).

Saunders, A. M., Albertsen, M., Vollertsen, J. & Nielsen, P. H. The activated sludge ecosystem contains a core community of abundant organisms. ISME J. 10 , 11–20 (2015).

PubMed PubMed Central Google Scholar

Jenkins, D., Richard, M.G., & Daigger, G.T. (ed. Jenkins, D.). Manual on the causes and control of activated sludge bulking, foaming, and other solids separation problems . 224 (CRC Press, 2003).

Henze, M., Harremoës, P., Jansen, L. C. & Arvin E. (eds. Henze, M., Harremoës, P., Jansen, L. C. & Arvin E) Wastewater treatment. Biological and chemical processe s. 383 (Springer, 1997).

Zhang, T., Shao, M. F. & Ye, L. 454 Pyrosequencing reveals bacterial diversity of activated sludge from 14 sewage treatment plants. ISME J. 6 , 1137–1147 (2012).

CAS PubMed Google Scholar

Wu, L. et al. Global diversity and biogeography of bacterial communities in wastewater treatment plants. Nat. Microbiol. 7 , 1183–1195 (2019).

Seviour, R.J. & Nielsen, P.H. (eds. Seviour, R.J. & Nielsen, P.H.) Microbial Ecology of Activated Sludge . 688 (IWA Publishing, 2010).

Nielsen, P. H., Daims, H., & Lemmer, H. (eds. Nielsen, P. H., Daims, H., & Lemmer, H.) FISH Handbook for Biological Wastewater Treatment . 200 (IWA Publishing, 2009).

Nierychlo, M. et al. MiDAS 3: an ecosystem-specific reference database, taxonomy and knowledge platform for activated sludge and anaerobic digesters reveals species-level microbiome composition of activated sludge. Water Res. 182 , 115955. https://doi.org/10.1016/j.watres.2020.115955 (2020).

Article CAS PubMed Google Scholar

Dueholm, M. S. et al. MiDAS 4: A global catalogue of full-length 16S rRNA gene sequences and taxonomy for studies of bacterial communities in wastewater treatment plants. bioRxiv. https://doi.org/10.1101/2021.07.06.451231 (2021).

Article Google Scholar

Kallistova, AIu. et al. Microbial composition of the activated sludges of the Moscow wastewater treatment plants. Microbiology 83 , 699–708 (2014) (In Russian) .

CAS Google Scholar

Shchegolkova, N. M. et al. Microbial community structure of activated sludge in treatment plants with different wastewater compositions. Front. Microbiol. 7 , 90 (2016).

Mardanov, A. V. et al. Metagenome of the microbial community of anammox granules in a nitritation/anammox wastewater treatment system. Genome Announc. 42 , e01115-e1117 (2017).

Mardanov, A. V. et al. Dynamics of the composition of a microbial consortium during start-up of a single-stage constant flow laboratory nitritation/anammox setup. Microbiology 85 , 681–692 (2016).

Nikolaev, Yu. A. et al. Novel design and optimisation of a nitritation/anammox set-up for ammonium removal from filtrate of digested sludge. Environ. Technol. 5 , 593–606 (2018).

Mardanov, A. V. et al. Genome of a novel bacterium “ Candidatus Jettenia ecosi” reconstructed from the metagenome of an anammox bioreactor. Front. Microbiol. 10 , 2442 (2019).

Danilovich, D. A. Blok udaleniya biogennyh elementov Lyubereckih ochistnyh sooruzhenij g. Moskvy–etapy vnedreniya sovremennyh tekhnologij. Nailuchshie dostupnye tekhnologii vodosnabzheniya i vodootvedeniya. 2 , 20–37 (2014) (In Russian) .

Danilovich, D. A. & Kozlov, M. N. 25 let promyshlennogo vnedreniya tekhnologij udaleniya azota i fosfora na moskovskih ochistnyh sooruzheniyah: 20 aprobirovannyh tekhnologicheskih reshenij. Nailuchshie dostupnye tekhnologii vodosnabzheniya i vodootvedeniya. 1 , 42–55 (2019) (In Russian) .

Kevbrina, M. V. et al. Anammoks–perspektivnaya tekhnologiya udaleniya azota iz stochnyh vod. Vodosnabzhenie i sanitarnaya tekhnika. 5 , 28–35 (2019) (In Russian) .

Kevbrina, M. V., Gavrilin, A. M., Dorofeev, A. G., Kozlov, M. N. & Aseeva, V. G. Nailuchshie dostupnye tekhnologii ochistki stochnyh vod: opyt vnedreniya AO «Mosvodokanal». Vodosnabzhenie i sanitarnaya tekhnika. 6 , 40–49 (2019) (In Russian) .

Frey, B. et al . Microbial diversity in European alpine permafrost and active layers. FEMS Microbiol Ecol . 3 , fiw018 (2016).

Magoc, T. & Salzberg, S. FLASH: Fast length adjustment of short reads to improve genome assemblies. Bioinformatics 27 , 2957–2963 (2011).

CAS PubMed PubMed Central Google Scholar

Edgar, R. C. Search and clustering orders of magnitude faster than BLAST. Bioinformatics 26 , 2460–2461 (2010).

Rognes, T., Flouri, T., Nichols, B., Quince, C. & Mahé, F. VSEARCH: a versatile open source tool for metagenomics. PeerJ. 4 , e2584 (2016).

Caporaso, J. et al. QIIME allows analysis of high-throughput community sequencing data. Nat. Methods. 7 , 335–336 (2010).

Yu, G., Lam, T. T. Y., Zhu, H. & Guan, Y. Two methods for mapping and visualizing associated data on phylogeny using ggtree. Mol. Biol. Evol. 2 , 3041–3043 (2018).

Yu, G., Smith, D. K., Zhu, H., Guan, Y. & Lam, T. T. Y. ggtree: an R package for visualization and annotation of phylogenetic trees with their covariates and other associated data. Methods Ecol. Evol. 1 , 28–36. https://doi.org/10.1111/2041-210X.12628 (2017).

Langfelder, P. & Horvath, S. Fast R functions for robust correlations and hierarchical clustering. J. Stat. Softw. 46 , i11 (2012).

Barberan, A. et al. Why are some microbes more ubiquitous than others? Predicting the habitat breadth of soil bacteria. Ecol. Lett. 17 , 794–802 (2014).

PubMed Google Scholar

Luo, X. & Bhattacharya, C. B. Corporate social responsibility, customer satisfaction, and market value. J. Mark. 70 , 1–18 (2006).

Shannon, P. et al. Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res. 11 , 2498–2504 (2003).

Faust, K. & Raes, J. CoNet app: inference of biological association networks using Cytoscape. F1000Res. 5 , 1519 (2016).

Parks, D. H. et al. A standardized bacterial taxonomy based on genome phylogeny substantially revises the tree of life. Nat. Biotechnol. 36 , 996–1004 (2018).

Dombrowski, N., Lee, J. H., Williams, T. A., Offre, P. & Spang, A. Genomic diversity, lifestyles and evolutionary origins of DPANN archaea. FEMS Microbiol Lett. 366 , fnz008 (2019).

CAS PubMed Central Google Scholar

van Kessel, M. A. et al. Complete nitrification by a single microorganism. Nature 528 , 555–559 (2015).

Lücker, S. et al. Nitrotoga-like bacteria are previously unrecognized key nitrite oxidizers in full-scale wastewater treatment plants. ISME J. 9 , 708–720 (2015).

Kitzinger, K. et al. Characterization of the first “ Candidatus nitrotoga” isolate reveals metabolic versatility and separate evolution of widespread nitrite-oxidizing bacteria. mBio. 9 , e01186-18 (2018).

Zheng, M. et al. Critical factors facilitating Candidatus nitrotoga to be prevalent nitrite-oxidizing bacteria in activated sludge. Environ Sci Technol. 54 , 15414–15423 (2020).

Van Loosdrecht, M. C. M., Hooijmans, C. M., Brdjanovic, D. & Heijnen, J. J. Biological phosphate removal processes. Appl. Microbiol. Biotechnol. 48 , 289–296 (1997).

Seviour, R. J., Mino, T. & Onuki, M. The microbiology of biological phosphorus removal in activated sludge systems. FEMS Microbiol. Rev. 27 , 99–127 (2003).

Dorofeev, A. G., Nikolaev, Y. A., Mardanov, A. V. & Pimenov, N. V. Cyclic metabolism as a mode of microbial existence. Microbiology 88 , 402–415 (2019).

Akbari, A. et al. Unrevealed roles of polyphosphate-accumulating microorganisms. Microb. Biotechnol. 14 , 82–87 (2021).

Hesselmann, R. P. X., Werlen, C., Hahn, D., van der Meer, J. R. & Zehnder, A. J. B. Enrichment, phylogenetic analysis and detection of a bacterium that performs enhanced biological phosphate removal in activated sludge. Syst. Appl. Microbiol. 22 , 454–465 (1999).

Stockholm-Bjerregaard, M. et al. Critical assessment of the microorganisms proposed to be important to enhanced biological phosphorus removal in full-scale wastewater treatment systems. Front. Microbiol. 8 , 718 (2017).

Zeng, W., Bai, X., Guo, Y., Li, N. & Peng, Y. Interaction of “ Candidatus Accumulibacter” and nitrifying bacteria to achieve energy-efficient denitrifying phosphorus removal via nitrite pathway from sewage. Enzyme. Microb. Technol. 105 , 1–8 (2017).

Qiu, G. et al. Polyphosphate-accumulating organisms in full-scale tropical wastewater treatment plants use diverse carbon sources. Water Res. 149 , 496–510 (2019).

Mino, T., van Loosdrecht, M. C. M. & Heijnen, J. J. Microbiology and biochemistry of the enhanced biological phosphate removal process. Water Res. 32 , 3193–3207 (1998).

Kong, Y., Xia, Y., Nielsen, J. L. & Nielsen, P. H. Structure and function of the microbial community in a full-scale enhanced biological phosphorus removal plant. Microbiology (SGM). 153 , 4061–4073 (2007).

Terashima, M. et al. Culture-dependent and -independent identification of polyphosphate-accumulating Dechloromonas spp. Predominating in a full-scale oxidation ditch wastewater treatment plant. Microbes. Environ. 31 , 449–455 (2016).

Shao, Y. et al. Zoogloea caeni sp. nov., a floc-forming bacterium isolated from activated sludge. Int. J. Syst. Evol. Microbiol. 59 , 526–30 (2009).

Pelevina, A. V. et al. A microbial consortium removing phosphates under conditions of cyclic aerobic-anaerobic cultivation. Microbiology 90 , 66–77 (2021).

Wang, Q. & He, J. Complete nitrogen removal via simultaneous nitrification and denitrification by a novel phosphate accumulating Thauera sp. strain SND5. Water Res. 185 , 116300 (2020).

Rubio-Rincón, F. J. et al. Long-term effects of sulphide on the enhanced biological removal of phosphorus: The symbiotic role of Thiothrix caldifontis . Water Res. 116 , 53–64 (2017).

Nguyen, H. T., Nielsen, J. L. & Nielsen, P. H. “Candidatus Halomonas phosphatis”, a novel polyphosphate-accumulating organism in full-scale enhanced biological phosphorus removal plants. Environ Microbiol. 10 , 2826–2837 (2012).

Spring, S., Wagner, M., Schumann, P. & Kampfer, P. Malikia granosa gen. nov., sp. nov., a novel polyhydroxyalkanoate- and polyphosphate-accumulating bacterium isolated from activated sludge, and reclassification of Pseudomonas spinosa as Malikia spinosa comb. nov.. Int. J. Syst. Evol. Microbiol. 55 , 621–629 (2005).

Maszenan, A. M. et al. Three isolates of novel polyphosphate-accumulating Gram-positive cocci, obtained from activated sludge, belong to a new genus, Tetrasphaera gen. nov., and description of two new species, Tetrasphaera japonica sp. nov. and Tetrasphaera australiensis sp. nov.. Int. J. Syst. Evol. Microbiol. 50 , 593–603 (2000).

Nielsen, J. L., Nguyen, H., Meyer, R. L. & Nielsen, P. H. Identification of glucose-fermenting bacteria in a full-scale enhanced biological phosphorus removal plant by stable isotope probing. Microbiology (SGM). 158 , 1818–1825 (2012).

Nakamura, K. et al. Microlunatus phosphovorus gen. nov., sp. nov., a new gram-positive polyphosphate-accumulating bacterium isolated from activated sludge. Int. J. Syst. Bacteriol. 45 , 17–22 (1995).

Wang, J. et al. The potential role of “Candidatus Microthrix parvicella” in phosphorus removal during sludge bulking in two full-scale enhanced biological phosphorus removal plants. Water Sci. Technol. 70 , 367–375 (2014).

Zhang, H. et al. Gemmatimonas aurantiaca gen. nov., sp. nov., a Gram-negative, aerobic, polyphosphate-accumulating microorganism, the first cultured representative of the new bacterial phylum Gemmatimonadetes phyl. nov.. Int. J. Syst. Evol. Microbiol. 53 , 1155–1163 (2003).

Soo, R. M. et al. An expanded genomic representation of the phylum Cyanobacteria . Genome Biol. Evol. 6 , 1031–1045 (2014).

Chen, H., Wang, M. & Chang, S. Disentangling community structure of ecological system in activated sludge: core communities, functionality, and functional redundancy. Microb. Ecol. 80 , 296–308 (2020).

Mardanov, A. V. et al. Genomic and metabolic insights into two novel Thiothrix species from enhanced biological phosphorus removal systems. Microorganisms. 8 (12), 2030 (2020).

Liu, R., Hao, X., Chen, Q. & Li, J. Research advances of Tetrasphaera in enhanced biological phosphorus removal: a review. Water Res. 166 , 115003. https://doi.org/10.1016/j.watres.2019.115003 (2019).

Rabus, R. Functional genomics of an anaerobic aromatic-degrading denitrifying bacterium, strain EbN1. Appl. Microbiol. Biotechnol. 68 , 580–587 (2005).

Oehmen, A. et al. Advances in enhanced biological phosphorus removal: from micro to macro scale. Water Res. 41 , 2271–2300 (2007).

Crocetti, G. R., Banfield, J. F., Keller, J., Bond, P. L. & Blackall, L. L. Glycogen-accumulating organisms in laboratory-scale and full-scale wastewater treatment processes. Microbiology (SGM). 148 , 3353–3364 (2002).

Wong, M. T., Tan, F. M., Ng, W. J. & Liu, W. T. Identification and occurrence of tetrad-forming Alphaproteobacteria in anaerobic-aerobic activated sludge processes. Microbiology 150 , 3741–3748 (2004).

Schramm, A. et al. On the occurrence of anoxic microniches, denitrification, and sulfate reduction in aerated activated sludge. Appl. Environ. Microbiol. 65 , 4189–4196 (1999).

ADS CAS PubMed PubMed Central Google Scholar

Rossetti, S., Tomei, M. C., Nielsen, P. H. & Tandoi, V. “Microthrix parvicella”, a filamentous bacterium causing bulking and foaming in activated sludge systems: a review of current knowledge. FEMS Microbiol. Rev. 29 , 49–64 (2005).

Fan, N. S., Qi, R., Huang, B. C., Jin, R. C. & Yang, M. Factors influencing Candidatus Microthrix parvicella growth and specific filamentous bulking control: a review. Chemosphere. 244 , 125371 (2020).

Nierychlo, M. et al. Low global diversity of Candidatus Microthrix, a troublesome filamentous organism in full-scale WWTPs. Front. Microbiol. 12 , 690251. https://doi.org/10.3389/fmicb.2021.690251 (2021).

Article PubMed PubMed Central Google Scholar

Strepis, N. et al. Genome-guided analysis allows the identification of novel physiological traits in Trichococcus species. BMC Genom. 21 , 24 (2020).

Chen, W. M., Yang, S. H., Huang, W. C., Cheng, C. Y. & Sheu, S. Y. Chitinivorax tropicus gen. nov., sp. nov., a chitinolytic bacterium isolated from a freshwater lake. Int. J. Syst. Evol. Microbiol. 62 , 1086–1091 (2012).

McIlroy, S. J. et al. Identification of active denitrifiers in full-scale nutrient removal wastewater treatment systems. Environ. Microbiol. 18 , 50–64 (2016).

Lu, H. P., Shao, Y. H., Wu, J. H. & Hsieh, C. H. System performance corresponding to bacterial community succession after a disturbance in an autotrophic nitrogen removal bioreactor. mSystems. 5 , e00398-20 (2020).

Iannacone, F., Di, C. F., Granata, F., Gargano, R. & Esposito, G. Simultaneous nitrification, denitrification and phosphorus removal in a continuous-flow moving bed biofilm reactor alternating microaerobic and aerobic conditions. Bioresour. Technol. 310 , 123453 (2020).

LaPara, T. M., Nakatsu, C. H., Pantea, L. & Alleman, J. E. Phylogenetic analysis of bacterial communities in mesophilic and thermophilic bioreactors treating pharmaceutical wastewater. Appl. Environ. Microbiol. 66 , 3951–3959 (2000).

Huang, X. et al. Sludge alkaline fermentation enhanced anaerobic- multistage anaerobic/oxic (A-MAO) process to treat low C/N municipal wastewater: nutrients removal and microbial metabolic characteristics. Bioresour. Technol. 302 , 122583 (2020).

Orsi, W. D. et al. Diverse, uncultivated bacteria and archaea underlying the cycling of dissolved protein in the ocean. ISME J. 10 , 2158–2173 (2016).

Speth, D. R., In ‘t Zandt, M. H., Guerrero-Cruz, S., Dutilh, B. E. & Jetten, M. S. Genome-based microbial ecology of anammox granules in a full-scale wastewater treatment system. Nat. Commun. 7 , 11172 (2016).

Franzmann, P. D. & Skerman, V. B. D. Agitococcus lubricus gen. nov., sp. nov., a lipolytic, twitching coccus from freshwater. Int. J. Syst. Bacteriol. 31 , 177–183 (1981).

Berry, D. & Widder, S. Deciphering microbial interactions and detecting keystone species with co-occurrence networks. Front. Microbiol. 5 , 219 (2014).

Kadnikov, V. V. et al. Phylogeny and physiology of candidate phylum BRC1 inferred from the first complete metagenome-assembled genome obtained from deep subsurface aquifer. Syst. Appl. Microbiol. 42 , 67–76 (2019).

Nielsen, P. H., Roslev, P., Dueholm, T. E. & Nielsen, J. L. Microthrix parvicella , a specialized lipid consumer in anaerobic-aerobic activated sludge plants. Water Sci. Technol. 46 , 73–80 (2002).

Klindworth, A. et al. Evaluation of general 16S ribosomal RNA gene PCR primers for classical and next-generation sequencing-based diversity studies. Nucleic Acids Res. 41 , e1 (2012).

Wasimuddin, et al. Evaluation of primer pairs for microbiome profiling from soils to humans within the One Health framework. Mol. Ecol. Resour. 6 , 1558–1571 (2020).

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This work was partly supported by the Russian Science Foundation (Project 21-64-00019 to A.V.M.).

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A.V.M. and N.V.R. designed and supervised the research project; A.G.D. collected the samples and analysed chemical composition of wastewater; V.V.K. and A.V.M. performed 16S rRNA gene profiling; S.B., A.V.B., N.V.P., and N.V.R. analysed the sequencing data; S.B. and N.V.R. wrote the manuscript. All authors have read and agreed to the published version of the manuscript.

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Begmatov, S., Dorofeev, A.G., Kadnikov, V.V. et al. The structure of microbial communities of activated sludge of large-scale wastewater treatment plants in the city of Moscow. Sci Rep 12 , 3458 (2022). https://doi.org/10.1038/s41598-022-07132-4

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Vietnamese dairy company TH Milk has broken ground on a new milk processing facility in Russia as part of its continued investment in the country. The plant, which will have a capacity of 1,500 ...
Exam 1 mkt mngm Flashcards
The research and development team of Nature's Way Beverages has discovered that, during the testing process, one of their profitable new beverages contains ingredients that may cause liver damage in humans if consumed regularly over time. ... Based on these findings, management has halted its production of this beverage until a solution can be ...
The structure of microbial communities of activated sludge of large
Microbial communities in wastewater treatment plants (WWTPs) play a key role in water purification. Microbial communities of activated sludge (AS) vary extensively based on plant operating ...

SBF's R&D Strength

Global Shift in Consumer Preferences toward Natural and Healthy Products

Research and Development— The Lifeblood of a Manufacturer

Co-creation of Global Value

Globalized product development

Teruyoshi Morikawa

Role and Function of the Research & Development within the MONOZUKURI Division

Creating a Unique Global Soft Drink Business Built on Local Autonomy

Recent Achievements Resulting from Global Synergies

Advantages in the Global Market

Developing products to satisfy the needs and preferences of local consumers

R&D Features Subscribe

Sports recovery, bone and joint health beverages appeal to all consumers, not just athletes

Beverage-makers turn to natural sweeteners as consumers aim to lower sugar intake

Beverage-makers work to provide cognitive support, energy

Weight management, sugar reduction trends transform beverage landscape

Beverage-makers explore ways to deliver on fortification trends

Natural colors help beverage-makers break from conventional

Consumers prefer performance beverages with qualities beyond hydration

IFT FIRST exemplifies innovations for beverage-makers

Consumers prefer natural, clean label beverage sweeteners

Consumers value beverages that support immune health

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Beverage Research and Development

How to Get Started

Partnership with Nature’s Flavors

Bottling Beverages

Canning Drinks

Research & Development

Evolution of Food Fermentation Processes and the Use of Multi-Omics in Deciphering the Roles of the Microbiota

Young-Su Seo

1. Introduction

2. Classification of Major Types of Fermented Foods and Beverages

3. Evolution of the Process of Fermentation over the Years

3.2. Starter Cultures

3.3. Starter Cultures of Multiple Strains and Adaptation for Co-Existence

3.4. Genetic Improvement of Starter Cultures

3.5. CRISPR/Cas9 Technology for Genetic Improvement of Starter Cultures

3.6. CRISPR-Mediated Microbiome Engineering and Fermentation

4. Multi-Omics and Microbiota Dynamics of Food Fermentation

4.1. Examples on the Use of Meta-Omics to Study Microbial Dynamics of Food Fermentation in Recent Studies

4.2. Functional Activity of the Food Fermentation Microbiome

5. Future Perspectives

Author Contributions

Conflicts of Interest

T. Hasegawa expands R&D with new division, leadership

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New products from Natural Products Expo West, part 1

New products from Natural Products Expo West, part 2

Reimagining Design with Nature: ecological urbanism in Moscow

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The politics of designing with nature: reflections from New Orleans and Dhaka

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The Role of Environmental NGOs: Russian Challenges, American Lessons: Proceedings of a Workshop (2001)

PRESENT SITUATION WITH WASTE IN MOSCOW

HOW TO CHANGE THE SITUATION WITH WASTE

ACTIVITY OF PUBLIC ORGANIZATIONS IN THE SPHERE OF WASTE MANAGEMENT IN THE MOSCOW REGION

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TH Milk breaks ground on $630m milk processing plant in Russia

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