Biopreservation

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The small rods shown here are lactic acid bacteria which convert lactose and other sugars to lactic acid. The products of their metabolism can have benign preservative effects. Lactobacillus sp 01.png
The small rods shown here are lactic acid bacteria which convert lactose and other sugars to lactic acid. The products of their metabolism can have benign preservative effects.
3D stick model of nisin, a particularly effective preservative produced by some lactic acid bacteria. Nisin 1WCO.png
3D stick model of nisin, a particularly effective preservative produced by some lactic acid bacteria.

Biopreservation is the use of natural or controlled microbiota or antimicrobials as a way of preserving food and extending its shelf life. [1] The biopreservation of food, especially utilizing lactic acid bacteria (LAB) that are inhibitory to food spoilage microbes, has been practiced since early ages, at first unconsciously but eventually with an increasingly robust scientific foundation. [2] Beneficial bacteria or the fermentation products produced by these bacteria are used in biopreservation to control spoilage and render pathogens inactive in food. [3] There are a various modes of action through which microorganisms can interfere with the growth of others such as organic acid production, resulting in a reduction of pH and the antimicrobial activity of the un-dissociated acid molecules, a wide variety of small inhibitory molecules including hydrogen peroxide, etc. [2] It is a benign ecological approach which is gaining increasing attention. [1]

Contents

Biopreservative agents and modes of action

Lactic acid bacteria

Of special interest are lactic acid bacteria (LAB). Lactic acid bacteria have antagonistic properties which make them particularly useful as biopreservatives. When LABs compete for nutrients, their metabolites often include active antimicrobials such as lactic and acetic acid, hydrogen peroxide, and peptide bacteriocins. Some LABs produce the antimicrobial nisin which is a particularly effective preservative. [4] [5]

These days LAB bacteriocins are used as an integral part of hurdle technology. Using them in combination with other preservative techniques can effectively control spoilage bacteria and other pathogens, and can inhibit the activities of a wide spectrum of organisms, including inherently resistant Gram-negative bacteria." [1] Lactic acid bacteria and propionibacteria have been extensively studied for their efficiency against spoilage causing yeasts and molds in food spoilage. [6]

This figure illustrates the pathway of food preservation followed by lactic acid bacteria involving nisin, as well as the pathway of food preservation followed by salt. Additionally, the hurdle effect of food preservation, such as by adding lactic acid bacteria and salt to a food product, is illustrated and described. Food Preservation by Lactic Acid Bacteria and Salt and The Hurdle Effect.png
This figure illustrates the pathway of food preservation followed by lactic acid bacteria involving nisin, as well as the pathway of food preservation followed by salt. Additionally, the hurdle effect of food preservation, such as by adding lactic acid bacteria and salt to a food product, is illustrated and described.

Yeast

In addition to lactic acid bacteria, yeasts also have been reported to have a biopreservation effect due to their antagonistic activities relying on the competition for nutrients, production and tolerance of high concentrations of ethanol, as well as the synthesis of a large class of antimicrobial compounds exhibiting large spectrum of activity against food spoilage microorganisms, but also against plant, animal and human pathogen. [11]

A bacterium/yeast that is a suitable candidate for use as a biopreservative does not necessarily have to ferment the food. However, if conditions are suitable for microbial growth, then a biopreservative bacterium will compete well for nutrients with the spoilage and pathogenic bacteria in the food. As a product of its metabolism, it should also produce acids and other antimicrobial agents, particularly bacteriocins. Biopreservative bacteria, such as lactic acid bacteria, must be harmless to humans. [3]

Bacteriophages

Bacteriophages (Greek for 'bacteria eater'), or simply phages, are viruses which infect bacteria. [12] The majority of all bacteriophages known exhibit a double-stranded DNA genome inside the virion capsid and belong to the order of tailed phages, Caudovirales. The tailed phages can be further separated into three families: Podoviridae, which are characterized by very short tails; Myoviridae, which exhibit longer, straight and contractile tails; and Siphoviridae, which can be identified due to their long and flexible tails. Another well studied group of phages with many applications, although minor in terms of species diversity, is represented by filamentous phages which exhibit a single stranded DNA genome decorated by a helical protein layer surrounding the DNA molecule. [12] Bacteriophages are ubiquitously distributed in nature and can also be isolated from human or animal associated microflora. They outnumber their bacterial host species by a factor of ten representing the most abundant self-replicating entities on earth with an estimated 1031 phages in total. [13] The idea of using phages against unwanted bacteria developed shortly after their discovery. With the improvements in organic chemistry during the 1950s, exploration and development of broad spectrum antibiotics displaced interest in bacteriophage research. Several laboratories have been testing suitability of bacteriophage isolates to control certain bacterial pathogens. Significant advancements in this research have been made at the Bacteriophage Institute in Tbilisi, Georgia, where phage therapy is routinely applied in medicine research field. Today treatment of antibiotic resistant bacteria is a challenging task. Recently, research on bacteriophages has gained additional momentum in light of the identification of antibiotic-resistant pathogens of infectious diseases, wherein the application of antibiotics is not effectively working, therefore research on the application of bacteriophages is being reviewed intensely. [12] Bacteriophages have recently received a generally recognized as safe status based on their lack of toxicity and other detrimental effects to human health for application in meat products in USA. [14]

Phage preparations specific for L. monocytogenes,E. coli O157:H7, and S. enterica serotypes have been commercialized and approved for application in foods or as part of surface decontamination protocols. [14]

Meat biopreservation

In meat processing, biopreservation has been extensively studied in fermented meat products and ready to eat meat products. [15] [16] [17] The use of native or artificially-introduced microbial population to improve animal health and productivity, and/or to reduce pathogenic organisms, has been termed a probiotic or competitive enhancement approach. [18] Competitive enhancement strategies that have been developed include competitive exclusion, addition of a microbial supplement (probiotic) that improves gastrointestinal health, and adding a limiting, non-host digestible nutrient (prebiotic) that provides an existing (or introduced) commensal microbial population a competitive advantage in the gastrointestinal tract. [16] Each of these approaches utilizes the activities of the native microbial ecosystem against pathogens by capitalizing on the natural microbial competition. Generally speaking, competitive enhancement strategies offer a natural 'green' method to reduce pathogens in the gut of food animals. [16]

Seafood biopreservation

Fishery products are a source of wide variety of valuable nutrients such as proteins, vitamins, minerals, omega-3 fatty acids, taurine, etc. Fishery products, however, are also associated with human intoxication and infection. Approximately 10 to 20% of food-borne illnesses are attributed to fish consumption. [19] Changing consumer demand has driven the appeal of traditional processes applied to seafood (e.g. salting, smoking and canning) lower compared to mild technologies involving lower salt content, lower cooking temperature and vacuum packing (VP)/modified atmosphere packing (MAP). These products, designed as lightly preserved fish products (LPFP), are usually produced from fresh seafood and further processing increases risk of cross contamination. [19] These milder treatments are usually not sufficient to destroy microorganisms, and in some cases psychrotolerant pathogenic and spoilage bacteria can develop during the extended shelf-life of LPFP. Many of these products are also eaten raw, so minimizing the presence and preventing growth of microorganisms is essential for the food quality and safety. [19] The microbial safety and stability of food are based on an application of preservative factors called hurdles. [20] The delicate texture and flavor of seafood are very sensitive to the decontamination technologies such as cooking, and more recent mild technologies such as pulsed light, high pressure, ozone, and ultrasound. Chemical preservatives, which are not processes but ingredients, are out of favor with consumers due to natural preservatives demand. An alternative solution that is gaining more and more attention is biopreservation technology. [20] [21] [22] In fish processing, biopreservation is achieved by adding antimicrobials or by increasing the acidity of the fish muscle. Most bacteria stop multiplying when the pH is less than 4.5. [19] Traditionally, acidity has been increased by fermentation, marination or by directly adding acetic, citric or lactic acid to food products. Other preservatives include nitrites, sulphites, sorbates, benzoates and essential oils. [4] The main reason for less documented studies for application of protective microorganisms, bacteriophages or bacteriocins on seafood products for biopreservation compared to dairy or meat products is probably that the early stages of biopreservation have occurred mainly in fermented foodstuffs that are not so developed among seafood products. [19] The selection of potential protective bacteria in seafood products is challenging due to the fact that they need adaptation to the seafood matrix (poor in sugar and their metabolic activities should not change the initial characteristics of the product, i.e. by acidification, and not induce spoilage that could lead to a sensory rejection. [19] Among the microbiota identified in fresh or processed seafood, LAB remains the category that offers the highest potential for direct application as a bioprotective culture or for bacteriocin production. [19]

Commercial applications and products

There has been successful implementation of various phage preparation around the globe. Various applications/delivery methods in food have been developed. Bacteriophages and their endolysins can be incorporated into food systems in several ways such as spraying, dipping or immobilization, singly or in combination with other hurdles. [23] The phage preparation LMP-1O2 has been subsequently commercialized as "ListShield" Intralyx, Inc. It has been shown to be effective against 170 different strains of "L. monocytogenes", reducing significantly (10 to 1000-fold) the Listeria contamination when sprayed onto ready-to-eat foods, without changing the food general composition, taste, odor or color. [14] The Intralytix company has also commercialized phage-based antimicrobial preparations like SalmoFresh and SalmoLyse for controlling S. enterica. [24] SalmoFresh is prepared with a cocktail of naturally occurring lytic bacteriophages that selectively and specifically kill Salmonella, including strains belonging to the most common/highly pathogenic serotypes Typhimurium, Enteritidis, Heidelberg, Newport, Hadar, Kentucky and Thompson. According to the manufacturer, SalmoFresh is specifically designed for treating foods that are at high risk for "Salmonella" contamination. In particular, red meat and poultry can be treated prior to grinding for significant reductions in Salmonella contamination. SalmoLyse is a reformulated phage cocktail derived from SalmoFresh in which two of the six phages in the original cocktail have been replaced. [24] Additional bacteriophage preparations have been formulated and referenced to be used to reduce the microbial load of animals prior to slaughter and are commercially available from Omnilytics such as the BacWash product line against Salmonella Omnilytics. Another commercial application has been developed, Listex_ P100 by Micreos in The Netherlands and was granted generally recognized as safe (GRAS) status by the FDA and USDA for use in all food products. [23]

Another significant commercial bacteriophage application is ELICOSALI, a wide range of anti-Salmonella and "E. coli" phage cocktail, for treatment of agricultural products developed by Eliava Institute at Tbilisi, Republic of Georgia Eliava Institute. [14]

Safety

Biopreservation judiciously exploits the antimicrobial potential of naturally occurring microorganisms in food and/or their metabolites with a long history of safe use. Bacteriocins, bacteriophages and bacteriophage-encoded enzymes fall in this theory. The long and traditional role of Lactic acid bacteria on food and feed fermentations is the main factor related to the use of bacteriocins in biopreservation. LAB and their bacteriocins have been consumed unintentionally for ages, laying down a long history of safe use. Their antimicrobial spectrum of inhibition, bactericidal mode of action, relative tolerance to processing conditions (pH, NaCl, heat treatments) and the lack of toxicity towards eukaryotic cells enforces their role as biopreservatives in food. [25] The evaluation of any new antimicrobial actives is done in meat by USDA which relies on the GRAS assessment by FDA among other suitability data.

Related Research Articles

<span class="mw-page-title-main">Food preservation</span> Inhibition of microbial growth in food

Food preservation includes processes that make food more resistant to microorganism growth and slow the oxidation of fats. This slows down the decomposition and rancidification process. Food preservation may also include processes that inhibit visual deterioration, such as the enzymatic browning reaction in apples after they are cut during food preparation. By preserving food, food waste can be reduced, which is an important way to decrease production costs and increase the efficiency of food systems, improve food security and nutrition and contribute towards environmental sustainability. For instance, it can reduce the environmental impact of food production.

A preservative is a substance or a chemical that is added to products such as food products, beverages, pharmaceutical drugs, paints, biological samples, cosmetics, wood, and many other products to prevent decomposition by microbial growth or by undesirable chemical changes. In general, preservation is implemented in two modes, chemical and physical. Chemical preservation entails adding chemical compounds to the product. Physical preservation entails processes such as refrigeration or drying. Preservative food additives reduce the risk of foodborne infections, decrease microbial spoilage, and preserve fresh attributes and nutritional quality. Some physical techniques for food preservation include dehydration, UV-C radiation, freeze-drying, and refrigeration. Chemical preservation and physical preservation techniques are sometimes combined.

<span class="mw-page-title-main">Biofilm</span> Aggregation of bacteria or cells on a surface

A biofilm comprises any syntrophic consortium of microorganisms in which cells stick to each other and often also to a surface. These adherent cells become embedded within a slimy extracellular matrix that is composed of extracellular polymeric substances (EPSs). The cells within the biofilm produce the EPS components, which are typically a polymeric conglomeration of extracellular polysaccharides, proteins, lipids and DNA. Because they have three-dimensional structure and represent a community lifestyle for microorganisms, they have been metaphorically described as "cities for microbes".

<span class="mw-page-title-main">Bacteriocin</span> Class of bacterially produced peptide antibiotics

Bacteriocins are proteinaceous or peptidic toxins produced by bacteria to inhibit the growth of similar or closely related bacterial strain(s). They are similar to yeast and paramecium killing factors, and are structurally, functionally, and ecologically diverse. Applications of bacteriocins are being tested to assess their application as narrow-spectrum antibiotics.

An organic acid is an organic compound with acidic properties. The most common organic acids are the carboxylic acids, whose acidity is associated with their carboxyl group –COOH. Sulfonic acids, containing the group –SO2OH, are relatively stronger acids. Alcohols, with –OH, can act as acids but they are usually very weak. The relative stability of the conjugate base of the acid determines its acidity. Other groups can [also] confer acidity, usually weakly: the thiol group –SH, the enol group, and the phenol group. In biological systems, organic compounds containing these groups are generally referred to as organic acids.

<i>Lactobacillus</i> Genus of bacteria

Lactobacillus is a genus of Gram-positive, aerotolerant anaerobes or microaerophilic, rod-shaped, non-spore-forming bacteria. Until 2020, the genus Lactobacillus comprised over 260 phylogenetically, ecologically, and metabolically diverse species; a taxonomic revision of the genus assigned lactobacilli to 25 genera.

<span class="mw-page-title-main">Nisin</span> Chemical compound

Nisin is a polycyclic antibacterial peptide produced by the bacterium Lactococcus lactis that is used as a food preservative. It has 34 amino acid residues, including the uncommon amino acids lanthionine (Lan), methyllanthionine (MeLan), didehydroalanine (Dha), and didehydroaminobutyric acid (Dhb). These unusual amino acids are introduced by posttranslational modification of the precursor peptide. In these reactions a ribosomally synthesized 57-mer is converted to the final peptide. The unsaturated amino acids originate from serine and threonine, and the enzyme-catalysed addition of cysteine residues to the didehydro amino acids result in the multiple (5) thioether bridges.

An antimicrobial is an agent that kills microorganisms (microbicide) or stops their growth. Antimicrobial medicines can be grouped according to the microorganisms they act primarily against. For example, antibiotics are used against bacteria, and antifungals are used against fungi. They can also be classified according to their function. The use of antimicrobial medicines to treat infection is known as antimicrobial chemotherapy, while the use of antimicrobial medicines to prevent infection is known as antimicrobial prophylaxis.

<i>Lactobacillus acidophilus</i> Species of bacterium

Lactobacillus acidophilus is a rod-shaped, Gram-positive, homofermentative, anaerobic microbe first isolated from infant feces in the year 1900. The species is most commonly found in humans,

<span class="mw-page-title-main">Fermented fish</span> Fish cured by fermentation to reduce spoilage

Fermented fish is a traditional preservation of fish. Before refrigeration, canning and other modern preservation techniques became available, fermenting was an important preservation method. Fish rapidly spoils, or goes rotten, unless some method is applied to stop the bacteria that produce the spoilage. Fermentation is a method which attacks the ability of microbials to spoil fish. It does this by making the fish muscle more acidic; bacteria usually cease multiplying when the pH drops below 4.5.

Industrial fermentation is the intentional use of fermentation in manufacturing processes. In addition to the mass production of fermented foods and drinks, industrial fermentation has widespread applications in chemical industry. Commodity chemicals, such as acetic acid, citric acid, and ethanol are made by fermentation. Moreover, nearly all commercially produced industrial enzymes, such as lipase, invertase and rennet, are made by fermentation with genetically modified microbes. In some cases, production of biomass itself is the objective, as is the case for single-cell proteins, baker's yeast, and starter cultures for lactic acid bacteria used in cheesemaking.

<span class="mw-page-title-main">Lactic acid bacteria</span> Order of bacteria

Lactobacillales are an order of gram-positive, low-GC, acid-tolerant, generally nonsporulating, nonrespiring, either rod-shaped (bacilli) or spherical (cocci) bacteria that share common metabolic and physiological characteristics. These bacteria, usually found in decomposing plants and milk products, produce lactic acid as the major metabolic end product of carbohydrate fermentation, giving them the common name lactic acid bacteria (LAB).

<span class="mw-page-title-main">Food microbiology</span> Study of the microorganisms that inhibit, create, or contaminate food

Food microbiology is the study of the microorganisms that inhabit, create, or contaminate food. This includes the study of microorganisms causing food spoilage; pathogens that may cause disease ; microbes used to produce fermented foods such as cheese, yogurt, bread, beer, and wine; and microbes with other useful roles, such as producing probiotics.

<span class="mw-page-title-main">Fish preservation</span>

Fish preservation is the method of increasing the shelf life of fish and other fish products by applying the principles of different branches of science in order to keep the fish, after it has landed, in a condition wholesome and fit for human consumption. Ancient methods of preserving fish included drying, salting, pickling and smoking. All of these techniques are still used today but the more modern techniques of freezing and canning have taken on a large importance.

<i>Pediococcus acidilactici</i> Species of bacterium

Pediococcus acidilactici is a species of Gram-positive cocci that is often found in pairs or tetrads. P. acidilactici is a homofermentative bacterium that can grow in a wide range of pH, temperature, and osmotic pressure, therefore being able to colonize the digestive tract. It has emerged as a potential probiotic that has shown promising results in animal and human experiments, though some of the results are limited. They are commonly found in fermented vegetables, fermented dairy products, and meat.

Hurdle technology is a method of ensuring that pathogens in food products can be eliminated or controlled. This means the food products will be safe for consumption, and their shelf life will be extended. Hurdle technology usually works by combining more than one approach. These approaches can be thought of as "hurdles" the pathogen has to overcome if it is to remain active in the food. The right combination of hurdles can ensure all pathogens are eliminated or rendered harmless in the final product.

<span class="mw-page-title-main">Reuterin</span> Chemical compound

Reuterin (3-hydroxypropionaldehyde) is the organic compound with the formula HOCH2CH2CHO. It is a bifunctional molecule, containing both a hydroxy and aldehyde functional groups.

<span class="mw-page-title-main">Food spoilage</span> Often due to bacteria and fungi

Food spoilage is the process where a food product becomes unsuitable to ingest by the consumer. The cause of such a process is due to many outside factors as a side-effect of the type of product it is, as well as how the product is packaged and stored. Due to food spoilage, one-third of the world's food produced for the consumption of humans is lost every year. Bacteria and various fungi are the cause of spoilage and can create serious consequences for the consumers, but there are preventive measures that can be taken.

Microbes can be damaged or killed by elements of their physical environment such as temperature, radiation, or exposure to chemicals; these effects can be exploited in efforts to control pathogens, often for the purpose of food safety.

Microbial food cultures are live bacteria, yeasts or moulds used in food production. Microbial food cultures carry out the fermentation process in foodstuffs. Used by humans since the Neolithic period fermentation helps to preserve perishable foods and to improve their nutritional and organoleptic qualities. As of 1995, fermented food represented between one quarter and one third of food consumed in Central Europe. More than 260 different species of microbial food culture are identified and described for their beneficial use in fermented food products globally, showing the importance of their use.

References

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  2. 1 2 Lacroix, Christophe. (2011). Protective Cultures, Antimicrobial Metabolites and Bacteriophages for Food and Beverage Biopreservation - 1. Identifying New Protective Cultures and Culture Components for Food Biopreservation. Woodhead Publishing. Online version available at: http://app.knovel.com/hotlink/pdf/id:kt009273P1/protective-cultures-antimicrobial/identifying-new-protective
  3. 1 2 Yousef AE and Carolyn Carlstrom C (2003) Food microbiology: a laboratory manual Wiley, Page 226. ISBN   978-0-471-39105-0.
  4. 1 2 FAO: Preservation techniques Fisheries and aquaculture department, Rome. Updated 27 May 2005. Retrieved 14 March 2011.
  5. ↑ Alzamora, Stella; Tapia, Maria Soledad; López-Malo, Aurelio (2000). Minimally Processed Fruits and Vegetables: Fundamental aspects and applications. Springer. p. 266. ISBN   978-0-8342-1672-3.
  6. ↑ Lacroix, Christophe. (2011). Protective Cultures, Antimicrobial Metabolites and Bacteriophages for Food and Beverage Biopreservation - 2. Antifungal Lactic Acid Bacteria and Propionibacteria for Food Biopreservation. Woodhead Publishing. Online version available at: http://app.knovel.com/hotlink/pdf/id:kt009274A1/protective-cultures-antimicrobial/antifungal-lactic-acid
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  11. ↑ Muccilli, S.; Restuccia, C. (2015). "Bioprotective Role of Yeasts". Microorganisms. 3 (4): 588–611. doi: 10.3390/microorganisms3040588 . PMC   5023263 . PMID   27682107.
  12. 1 2 3 Lacroix, Christophe. (2011). Protective Cultures, Antimicrobial Metabolites and Bacteriophages for Food and Beverage Biopreservation - 6. Bacteriophages and Food Safety. Woodhead Publishing. Online version available at: http://app.knovel.com/hotlink/pdf/id:kt009277EL/protective-cultures-antimicrobial/bacteriophages-food-safety
  13. ↑ BRUSSOW H and KUTTER E (2005). 'Phage ecology.' In Kutter E and Sulakvelidze A (eds) Bacteriophages - Biology and Application, New York, CRC Press, 13 1 pp.
  14. 1 2 3 4 Pérez Pulido, Rubén; Grande Burgos, Maria José; Gálvez, Antonio; Lucas López, Rosario (2015). "Application of bacteriophages in post-harvest control of human pathogenic and food spoiling bacteria". Critical Reviews in Biotechnology. 36 (5): 1–11. doi:10.3109/07388551.2015.1049935. ISSN   0738-8551. PMID   26042353. S2CID   20081805.
  15. ↑ Lacroix, Christophe. (2011). Protective Cultures, Antimicrobial Metabolites and Bacteriophages for Food and Beverage Biopreservation - 7. Using Antimicrobial Cultures, Bacteriocins and Bacteriophages to Reduce Carriage of Food-Borne Bacterial Pathogens in Poultry. Woodhead Publishing. Online version available at: http://app.knovel.com/hotlink/pdf/id:kt009277U3/protective-cultures-antimicrobial/using-antimicrobial-cultures
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