VI.

BIOPRESERVATION/ BIOLOGICAL METHODS OF FOOD PRESERVATION Introduction Food preservation by using temperature, drying and chemicals is an old method of preservation of food. In temperature mediated preservation of food, the enzymes which are (promote self decomposition of food) present in the food as well as in the microbes (which cause spoilage) are targeted. In high temperature, the enzymes are denatured and in low temperature, they are inactivated so that spoilage by decay/spoilage by self decomposition and by microbes is avoided. In chemically preserved food, the chemicals which are added to the food have their own ill effects on human health even though this method provides for the diversity and safety of many food products. Among the new methods of food preservation, the role of radiation and the microbes especially the Lactic acid bacteria are gaining importance in food preservation. In case of irradiation mediated food preservation, the multiplication of microbes is arrested by ionizing the atoms/ molecules which is used for the formation of DNA of microbes. The following paragraphs describe the role biopreservation of food materials in detail. Definition: Biopreservation is one of the recent methods of food preservation. It refers to the use of microorganismsmainly bacteria, their metabolic products or both into various types of food substances to delay/avoid spoilage by the spoilage microbes and ensures quality and health benefits to the consumer. OR Biopreservation can also refer to the deliberate use of environmental conditions that cause a target (pathogenic microbes) microorganism to stop growing and become metabolically inert (inactive). The still-living (beneficial one) microbe then has a better chance of surviving inhospitable conditions that would kill their actively growing and dividing counterparts. An example of biopreservation is the deliberate introduction of bacteria from a group called lactic acid bacteria or their metabolic products or both into various food (fermented and nonfermented) products, including fish and vegetables. This form of biopreservation can be useful in storing commercially and medically important microorganisms in addition to food preservation. Evolution of the concept of biopreservation/ Scientific foundations of biopreservation The use of microorganisms as a means of minimizing the spoilage and contamination of foods has its origin in experiments conducted beginning in the 1980s that demonstrated the beneficial health effects of the deliberate introduction of bacteria in the body. This research showed that the introduction of bacteria called Lactobacillus could reduce the adherence of pathogenic (capable of causing disease) bacteria. Similarly, research has shown that a chemical ingredient in cranberry juice can block a site on the surface of intestinal cells that is used as an anchor for the binding of a pathogenic version of the bacteria Escherichia coli. The use of bacteria and other microorganisms as medical and nutritional aidsan approach called probiotics led to the use of bacteria as food biopreservatives. For example, growth of lactic acid bacteria in a food will lower its pH because of the production of the acid. The lower pH makes the food less hospitable for problematic bacteria, which slows their growth. The benefits of food biopreservation can be profound. As an example, ingesting food contaminated with Listeria monocytogenes can cause a disease called listeriosis. The disease kills 20 to 40 percent of those who become infected. Pregnant women, infants, the elderly, and people whose immune systems are not operating efficiently are particularly at risk from listeriosis. Significance or importance of biopreservation 1. The consumers are increasingly become wary of chemical preservatives used for food preservation because of their ill effects on human health. E.g. Nitrosamines are carcinogenic compound formed from nitrites when nitrites are used as curing agent in meat. 2. Instead of chemically preserved food, the consumer prefers to have minimally processed food. But the minimally processed food relies on the use of refrigeration for the storage safety (to avoid spoilage).Around 20% of the commercial refrigerators used to store this minimally processed food maintain >10 degree Celsius temperature which cannot ensure the food free from spoilage. Even in good working refrigerator, the pathogens such as Listeria monocytogenes and other pathogens can grow and spoil food at freezing temperature of the refrigerated storage. 3. Inability of traditional food preservation methods to eliminate the pathogen such as Listeria monocytogenes which is recalcitrant to most of the traditional food preservation methods and causes a food borne disease listeriosis. 4. Increasing awareness of the consumer to have not only the natural but also the health promoting food over the chemically preserved food. 5. Prevention of lactose intolerance and and diarrhea: 6. The consumption of large quantities of fermented dairy products decreases the serum cholesterol.

7. 8.

Prevention of colointestinal cancer: Biopreserved food is said to be free from pathogenic microbes such as Listeria monocytogenes, pathogenic version of E.coli and Clostridium botulinum etc. Antimicrobial proteins such as Nisin kill the spores and vegetative cells of Listeria and Staphylococcus by making them sensitive to heat. 9. Antimicrobial proteins (bacteriocins) such as Nisin has the potential to reduce the ulcers in human and it also acts as a general sanitizing agent in human. 10. LAB ensures improvement in the quality (by adding characteristic flavor) of fermented food when it is used as starter culture for fermented food. 11. Biopreservation is useful in cell/organ transplantation and can also be useful in storing commercially and medically important microorganisms.

a. Health promoting effects of biopreserved food: 1. Prevention of lactose intolerance and and diarrhea: The intestinal mucosa of the Asian and African descent deficient in Beta glycosidase activity which results in inability to metabolize the lactose in dairy products and causing flatulence, pain and diarrhea. Thus theses population is deprived of calcium and proteins. Addition of Lab starter culture to the dairy products to overcome these problems. The LAB may metabolize the lactose in the gut or it may stimulate the lactase production in the mucosa. The activity of LAB on diary products is limited to the only fermented dairy food products which contain lactose. Consuming LAB prevents and reduces the severity of diarrhea illness. 2. Reduction of serum cholesterol: The consumption of large quantities of fermented dairy products decreases the serum cholesterol. 3. reduction colon and intestinal cancer Many human cancers are triggered by chemicals or events in the environment. The development of breast, colon, and prostrate cancers is associated with diets that are high in fat and low in fiber. Most dietary carcinogens are consumed as procarcinogens which require chemical modification to induce carcinogenesis. The chemical modification is done by the enzymes of microbes present in the gastrointestinal tract and to small degree by the enzymes of the intestinal mucosa. The enzymatic reactions of the gastrointestinal tract are classified as either procarcinogenic (phase1) or anti carcinogenic reactions (phase11).Both (phase1) or (phase11) enzymes help the body excrete compounds rendering them less lipid soluble. Phase 1 reactions do this by hydrolysis, oxidation, or reduction of the parent compound. These reactions activate procarcinogens and are heavily influenced diet Cycasin. b. Role of biopreservation in cell/organ transplantation The ability to routinely transplant cells and even whole organs from one person to another could be greatly aided by the ability to store the cells and organs for longer periods of time. Currently, the only way to store tissues is at very low temperature, which can destroy some of their components such as proteins. Scientists are exploring the use of biopreservation of proteins and even entire organs using microorganisms that are resistant to desiccation (the removal of water). Transfer of a protein species to a desiccation-resistant microbe could preserve the protein; subsequently the protein could be extracted from the microbe for use in a transplant. Although it is currently in the experimental stages, biopreservation of transplantable materials may some day be a routine feature of medicine.

1. Lactic acid bacteria The lactic acid bacteria form a heterogeneous group of Gram-positive bacteria such as such as Lactobacillus, Lactococcus, Streptococcus, Pediococcus, and Leuconastoc and Oenococcus etc. They physiologically ferment carbohydrates (lactose) into lactic acid as the predominant metabolic end-product. Homofermentative LAB converts fermentable carbohydrates almost quantitatively (produce more than 85% lactic acid from glucose) into lactic acid whereas heterofermentative produce only 50% lactic acid and considerable

amounts of ethanol, acetic acid and carbon dioxide. Some LAB also produces flavor compounds that are essential to the final taste of the fermented products. Lactic acid bacteria are used in the food industry for several reasons. Their growth lowers both the carbohydrate content of the foods that they ferment, and the pH due to lactic acid production. It is this acidification process which is one of the most desirable side-effects of their growth. The pH may drop to as low as 4.0, low enough to inhibit the growth of most other microorganisms including the most common human pathogens,(E.g. Clostridium botulinum and pathogenic version of the bacteria Escherichia coli etc. ) allowing these foods to have prolonged shelf life. The acidity also changes the texture of the foods due to precipitation of some proteins. The fermentation (and growth of the bacteria) is self-limiting due to the sensitivity of lactic acid bacteria to such acidic pH. Lactic acid is reported to have some physiological benefits such as: a) Enhancing the digestibility of milk proteins by precipitating them in fine curd particles. b) Improving the utilization of calcium, phosphorus and iron. c) Stimulating the secretion of gastric juices d) Accelerating the onward movement of stomach contents e) Serving as a source of energy in the process of respiration. (E.g. Microgard is a commercialized fermented milk product added to the cottage cheese to preserve the cheese. The cheese is preserved because of the formation of acids such as propionic acid, acetate and low molecular weight proteins by the Propionibacterium shermani under temperature abusive conditions.) Lactose metabolism: Lactic acid bacteria have the enzymes b-glycosidase, glycolases and lactic dehydrogenase (LDH) which produce lactic acid from lactose. The lactic acid has two isomeric forms. The structural configurations of the lactic acid isomers are as follows:

D (-) levorotatory lactic acid

L (+) dextrorotatory lactic acid

In humans, both isomers are absorbed from the intestinal tract. Whereas L (+) lactic acid (L. acidophilus) is completely and rapidly metabolized in glycogen synthesis, D (-) lactic acid (L. sporogenes) is metabolized at a lesser rate, and the unmetabolized acid is excreted in the urine. The presence of unmetabolized lactic acid results in metabolic acidosis in infants. II. Antimicrobial compounds

Bacteriocins Hydrogen peroxide Carbon dioxide Diacetyl

Mechanism of action of different metabolites produced by Lactic Acid Bacteria The metabolites of lactic acid bacteria that exert antagonistic action against putrefactive microorganisms and their mode of action are summarized below.

Metabolic product 1.Carbon dioxide 2. Diacetyl 3. Hydrogen peroxide / Lactoperoxidase

Mode of antagonistic action on the pathogens Inhibits decarboxylation and reduces membrane permeability Interacts with arginine-binding proteins. Oxidizes basic proteins. Undissociated lactic acid penetrates the membranes, lowering the intracellular pH. It also interferes with metabolic processes such as oxidative phosphorylation. Affect membranes, DNA-synthesis and protein synthesis.

4. Lactic acid

5. Bacteriocins

Bacteriocins: Bacteriocins are heterogeneous group of ribosomal produced small proteins produced by LAB. They act against the closely related bacteria but not against the producing organism. In some cases closely related species covers the wide range of gram positive bacteria. The gram negative bacteria can be made sensitive to the bacteriocin in the presence of chelating agent, hydrostatic pressure, or injury. The bacteriocins act on the cell or protoplasmic membrane, inhibit protein synthesis, degrade DNA and RNA etc. of pathogenic microbes. The inhibitory activity of lactobacilli towards putrefactive organisms is thought to be partially due to the production of bacteriocins. Some of the bacteriocins isolated from lactobacilli are listed below. Bacteriocins isolated from different Genus and species of Bacteria. Substance Acidolin Acidophilin Lactacin B Lactacin F Bulgarin Plantaricin SIK-83 Plantaricin A Lactolin Plantaricin B Lactolin 27 Helveticin J and A Reuterin Lactobrevin Producing species L. acidophilus L. acidophilus L. acidophilus L. acidophilus L. bulgaricus L. plantarum L. plantarum L. plantarum L. plantarum L. helveticus L. helveticus L. reuteri L. brevis

Lactobacillin

L. brevis

Subtilin

Bacillus subtilis

Pediococcus sp Pediocin

? Nisin Leuconosin Leuconostoc sp

Characteristics of Bacteriocins: Most bacteriocins are small protein compounds having the molecular weight of 3-30 kD Have high isoelectric point (The isoelectric point (pI) is the pH at which a particular molecule or surface carries no net electrical charge. Amphoteric molecules called zwitterions contain both positive and negative charges depending on the functional groups present in the molecule. They are affected by pH of their surrounding + environment and can become more positively or negatively charged due to the loss or gain of protons (H ). They contain both hydrophobic and hydrophilic domains Their names based on the amino acid sequence of proteins and also the bacterial species from which they are obtained They act both against the gram positive and gram negative bacteria They act against the closely related bacteria but not against the producing organism. The bacteriocins act on the cell or protoplasmic membrane, inhibit protein synthesis, degrade DNA and RNA etc. of pathogenic microbes Types of bacteriocins Group-I bacteriocins: Group I Bactericins contain the unusual amino acids such as dehydro alanine, dehydrobutyrine, lanthionine, Beta methyl lanthione. These amino aids are produced by the post translational modification of serine and threonine to their dehydro forms. The dehydro amino acids react with cysteine to form thioether (single sulfhydry)l lanthionine rings. Bacteriocins containing lanthionine rings referred as lantibiotics. Nisin, subtilin, lacticin 481, lactococcin, lactocin and carnocin are the examples of lantibiotics produced by LAB. Group-II bacteriocins: Group-II is large group of heat stable proteins with a consensus leader sequence containing Gly-Gly-1-Xaa+1 cleavage site important for processing the prebacteriocin during the export. Xaa= any amino acid Sub groups of group-II bacteriocins: Sub group II a; Theses subgroup Bacteriocin has Tyr-Gly-Val-Xaa-Cys terminal sequence and act against Listeria monocytogenes. Sub group II b: E.g. Pediocin PA-I, Sakacin A and P, Leucocin A, Bavaricin MN and Curvacin A. Sub group II c: E.g. Lactacin B Group-III: E.g. Helveticin J and V

Group-IV : E.g. Leuconocin S, lactocin 27 and Pediocin-SJ-1

Group III and Group-IV Bacteriocins are large >30KD heat labile proteins which are markedly differ from other bacteriocins. (Protein Sequences

The one-letter and three-letter abbreviation codes for amino acids for example, used in UniProtKB/Swiss-Prot are those adopted by the commission on Biochemical Nomenclature of the IUPAC-IUB and are as follows: One-letter code A R N D C Q E G H J L I K M F P O U S T W Y V B Z X ) Ala Arg Asn Asp Cys Gln Glu Gly His Xle Leu ILe Lys Met Phe Pro Pyl Sec Ser Thr Trp Tyr Val Asx Glx Xaa Three-letter code Alanine Arginine Asparagine Aspartic acid Cysteine Glutamine Glutamic acid Glycine Histidine Leucine or Isoleucine Leucine Isoleucine Lysine Methionine Phenylalanine Proline Pyrrolysine Selenocysteine Serine Threonine Tryptophan Tyrosine Valine Aspartic acid or Asparagine Glutamic acid or Glutamine any amino acid Amino-acid name

Detailed mechanism of action of bacteriocins 1) Action against vegetative cells of pathogenic microbes LAB bacteriocins act at the cell membrane. They disrupt the integrity of cytoplasmic membrane, thus increasing its permeability to small compounds. Addition of bacteriocins to vegetative cells results in rapid and nonspecific efflux of pre accumulated ions, amino acids and in some cases ATP molecules from the cytoplasmic membrane of the vegetative cells due to membrane disruption. The proton motive force (PMF), an electrochemical gradient which serves as the driving force of much vital energy dependent processes is dissipated within minutes of bacteriocin addition. The chemical gradient component of the proton motive force is dissipated faster than the electrical component as a result of rapid influx of protons. The loss of ATP caused by efflux or ATP hydrolysis. Ultimately, these changes in permeability render the cell unable to protect its cytoplasm from the environment. This leads to cell inhibition and possibly death. The membrane disruption by bacteriocin is promoted by two different mechanisms. In the first one is the formation of multistep-step poration complex (pore formation) in which bacteriocin monomer bind with, insert, and oligomerise in the cytoplasmic membrane to form a pore with the hydrophilic residues facing inward and the hydrophobic ones facing outwards. Alternatively, the bacteriocins disrupt the membrane integrity by a detergent-like membrane solublization (membrane solubilization) action.

2) Action against spores of pathogen Information about the mechanism of action of bacteriocin against spores deals primarily with the Nisin. Spore germination and growth is a multistep process. Nisin allows the spores to germinate but inhibit the preemergent spores at high concentration when compared to vegetative cells. The spores are opened and killed by lysis by these Nisin. At molecular level, nisin modifies the s[ulfhydral group in the envelop of germinated spores. Method of application of antimicrobial compounds: One way of application of bacteriocins is that the metabolites (antimicrobial proteins such as Nisin and Pediocin) can be added directly to the food materials to inhibit the pathogenic microbes. Among the antimicrobial proteins, only Nisin is available commercially in pure form for the direct addition to food and for the Pediocin the demonstration has been done but it has to be commercialized. Another way of application is the addition of bacteriocinogenic culture (Bacteria producing bacteriocin) to the nonfermented food or used as a starter culture for the fermented foods. The addition of starter culture (bacteriocinogenic culture) in the beginning to the fermented food improves the quality of fermented food than the indigenous microflora by replacing /inactivating the indigenous microflora of the fermented food. E.g. In dairy products, the starter culture are added after pasteurization because pasteurization kills the indigenous microflora. Current issues on biopreservation and limitations in biopreservation research While biopreservation of food is a very promising approach to maximizing food quality, more research is needed to ensure that the lactic acid and other types of bacteria do not themselves cause as-yet unforeseen problems. The likelihood of this is fairly remote, since lactic acid bacteria have been studied for over a century. Regarding the bacteriophge, eventhough the FDA approval has been granted, limitations of the approach presently limit its usefulness. The fact that a given bacteriophage infects only one or a very limited number of types of pathogenic/spoilage bacteria. Another, much more problematic disadvantage, is the potential that a bacteriophage can act as a vector to transfer genes from one bacterium to other bacterium.These current limitations with the use of bacteriophages will need to be overcome if the approach is to become feasible for the safe, large-scale biopreservation of foods.

VII. PROBIOTICS, PREBIOTICS and SYNBIOTICS Introduction The purpose of food is to provide nutrition to all living oraganisms.The growing awareness of people to have healthy diet (diet which promotes the health in human body) has led to an increasing demand for food products that support health above and beyond providing basic nutrition. The food which ensures beneficial health effects plus basic nutrition to the human body is called functional food. The functional food can be obtained by incorporating the beneficial probiotics and prebiotics in to the food. While the benefits of probiotics have come to light in more recent years, recognition of probiotic effects dates back to the 19th century when the French scientist Louis Pasteur (18221895) postulated the importance of microorganisms in human life; this was further reinforced by work done by 1908 Nobel Prize-winner Elie Metchnikoff. 1.Probiotics: A probiotic has been defined as a live microbial food ingredient that, when ingested in sufficient quantities, exerts health benefits on the host.(Human/animals) Similarly, the Joint Working Group of Food and Agriculture Organization and World Health Organization (which drafts the Guidelines for the Evaluation of Probiotics in Food) has recommended that probiotics be defined as Live microorganisms which, when administered in adequate amounts, confer a health benefit on the host. A. Prebiotics, sources and health benefits Probiotics Certain species and strains of Lactobacilli, Bifidobacteria Source Certain yogurts other cultured dairy products Potential Benefit May improve gastrointestinal health and systemic immunity

Pediococcus, Streptococcus Aspergillus Bacillus And Yeast

and non-dairy applications

B.Specific requirement of Probiotics to ensure health benefits 1. Selection of suitable species and strains of microbes because the health benefits associated with probiotics are strain and food substance specific. 2. Adequate clinical trials should be done to fix the correct dose of pro- or prebiotics present in the food at the time of consumption. 3. After deciding the required dose or quantity by adequate clinical trials, the Probiotics and Prebiotics can be added as a supplement to the food. 4. The survival of the probiotics in the GI tract can be improved by microencapsulation of probiotics, adding prebiotics such as inulin and fructo-oligosacharides and adding other food additives such as vitamins. C. Characteristics of probiotics: Acid tolerance Bile tolerance Cell surface hydrophobicity Protoplast regeneration Antimicrobial activity Cholesterol removal and bile salt deconjugation Gut colonization Lactose removal Protease and aminopeptidase activity D. Role of Probiotics in human GI tract Human beings play host to many types of microorganisms on the skin, in the mouth and in the GI tract. The human gastrointestinal environment, including the microflora, has a significant role in the health of its host. The normal gut microflora activity is complex and can be impacted by a number of factors. The gut microflora, which includes both potentially beneficial and potentially harmful bacteria, is important in maintaining a healthy intestinal tract and helps the intestine act as an effective barrier; allowing nutrients to be absorbed, and keeping out toxins and pathogens (foreign bacteria or viruses). The gut microflora breaks down vitamins and also ferments fibers and carbohydrates that are not digested in the upper GI tract. This breakdown produces fatty acids that are important for supporting a healthy intestinal barrier (particularly in the lower GI tract) and also inhibits the growth of harmful bacteria. Healthy intestinal flora is also associated with intestinal (stool) regularity.

E. Negative effects of imbalance of microflora found in the human body or Problems can be overcome by probiotics in the human body

Infant allergies and infections (when used in formula) Inflammatory colitis Candida yeast infections (in all body areas) Atopic dermatitis Bladder infections Vaginal infections Acute pancreatitis Indigestion

Ulcerative colitis Antibiotic-induced diarrhea Gastroenteritis (stomach flu) Diarrhea Risk of colon cancer Gastric ulcers Duodenal ulcers Helicobacter pylori infections Post-operative bowel infections

High cholesterol Crohn's Disease Non-alcoholic fatty liver disease

Clostridium difficile infections Complications of prematurity Abdominal radiation

F. Health effects of Probiotics Perhaps the most compelling evidence for probiotic efficacy is in the areas of 1. Elimination of lactose intolerance and anti diarrheal effects The intestinal mucosa of the Asian and African descent deficient in Beta galactosidase activity which results in inability to metabolize the lactose in dairy products and causing flatulence, pain and diarrhea. Thus theses population is deprived of calcium and proteins. These problems due to lactose nonmetabolism can be overcome by addition of LAB starter culture (Streptococcus thermophilus and Lactobacillus bulgaricus) to the dairy products. The LAB may metabolize the lactose in the gut or it may stimulate the lactase or Beta galactosidase production in the mucosa. The activity of LAB on diary products is limited to the only fermented dairy food products which contain lactose. Consuming LAB prevents and reduces the severity of diarrhea illness. A study in young children showed a clear beneficial effect of the above mentioned two Lactobacillus strains on the improvement of acute rotavirus diarrhea. In adults there are limited data to support an effect on either travelers diarrhea or that caused by antibiotic use. 2. Enhancement of immune system: Some probiotic strains can modulate the immune system and enhance immune system defenses of the intestinal mucosal cells through both specific and non-specific effects. Certain probiotic strains have been shown to have a favorable effect on markers of the immune response to stress. A study among the elderly found an enhancement of immune function following consumption of milk supplemented with a Bifidobacterium lactis strain.

3. Health effects on infants: Bifidobacteria is the predominant bacterial species in the intestinal tracts of breast fed infants. Some experts believe higher levels of Bifidobacteria in the gut of breastfed infants may be one reason why they are considered to be generally healthier than formula-fed babies. 4. There is also a growing body of new evidence that some probiotic strains may have a beneficial impact on allergy in infants. 5. Some probiotic strains may also reduce the severity of pathogenic microbes-induced gut inflammation, acute gastroenteritis and inflammatory bowel disease. 6. While research is ongoing, current evidence with several probiotic strains in animal and human studies suggests a moderate cholesterol-lowering effect from cultured dairy products such as yogurt and milk products. 7. The probiotics have the effect of reducing the colorectall cancer through neutralizing the action of dietary carcinogens in the GI tract. 2. Prebiotics: Prebiotics are defined as nondigestible but fermentable carbohydrate (oligosaccharides) food ingredients that beneficially affect the host by selectively stimulating the growth of one or a limited number of bacterial species in the colon, such as Bifidobacteria and Lactobacilli etc. which have the potential to improve host health. Prebiotics are, simply speaking, the food for beneficial bacteria. The food ingredients should posses the following characterestics to be a prebiotic

(a) is not broken down in the stomach or absorbed in the GI tract, (b) is fermented by the gastrointestinal microflora; and (c) Most importantly, selectively stimulates the growth and/or activity of intestinal bacteria associated with health and wellbeing of the host. Prebiotics are found naturally in many foods, and can also be isolated from plants (e.g., chicory root) or synthesized (e.g., enzymatic ally, from sucrose). A. Prebiotics, sources and health benefits Prebiotics Source B. Health effects of prebiotics 1. The principal characteristic and effect of prebiotics in the diet is to promote the growth and proliferation of beneficial bacteria in the intestinal tract, and thus, potentially yield or enhance the effect of probiotic bacteria. 2. Prebiotics have also been shown to increase the absorption of certain minerals (such as calcium and magnesium). Recent human studies have confirmed that specific prebiotics, such as non-digestible oligosaccharides (NDOs) enhance calcium absorption. However, this effect varies according to the individual NDO and particular human population studied, and is thought to be due to its specific fermentation profile and the amount consumed. 3. Prebiotics may also help inhibit the growth of lesions, such as adenomas and carcinomas in the gut, and thus reduce the risk factors involved in colorectal diseases. 4. Preliminary studies suggest that prebiotics may have a favorable effect on the immune system and provide improved resistance against infection. 5. For prebiotic substances, little data pertaining to lipid-lowering effects are available and come mostly from studies with inulin and oligofructose. 6. In hyperlipidemic subjects, when a prebiotic effect is seen, it is a reduction in cholesterol; whereas in normallipidemic subjects, any noted effects are on serum triglycerides. 7. Some of the pro- and prebiotic beneficial effects on the function of the human gut have been established and their favorable impact on health widely supported, further scientific research is ongoing to substantiate their direct relationship to disease risk reduction. 3. Synbiotics: Probiotic bacteria taken together with prebiotics that support the growth of beneficial bacteria are called synbiotics. Both work together in a synergistic way more efficiently promoting the probiotics benefits. Whole grains onions bananas garlic honey Leeks artichokes fortified foods and beverages dietary supplements and other food applications Potential Benefit

Inulin Fructo-oligosaccharides (FOS) Polydextrose Arabinogalactan Polyolslactulose and Lactitol etc.

May improve gastrointestinal health may improve calcium absorption

The bottom line The healthful effects of pre- and probiotics factor in their potential impact on the balance of the bodys microflora, and directly or indirectly in their enhancement of the function of the gut and systemic immune system. Although benefits vary, depending on the type and amount of a pre- or probiotic consumed, experts agree that daily consumption of foods containing these functional components is beneficial. In addition, effects of probiotics are strain-specific and must be demonstrated through appropriate clinical trials.