1. Probiotics, Biotechnology and Health
Present and Future
Mtro. Pedro Javier Guerrero Medina
Febrero 2012

2. Content
History
Definitions
Present
Future
Conclusions

3. History Metchnikoff, (1907) first introduced the probiotics concept.
Tissier, (1906) “bifid” bacteria abundant in healthy children.
Lilly and Stillwell, "probiotic“ to name substances produced by microorganisms which promoted the growth of other microorganisms
Fuller (1989), "A live microbial feed supplement which beneficially affects the host animal by improving its intestinal balance".
Havenaar and Huis in 't Veld (1992), “a viable mono or mixed culture of bacteria which, when applied to animal or man, beneficially affects the host by improving the properties of the indigenous flora”
Guarner and Schaafsma, (1998), "live microorganisms, which when consumed in adequate amounts, confer a health effect on the host”

4. Some of the descriptions and definitions of probiotics commonly cited over the years
Source
Probiotics are common in vegetable food as vitamins aromatic substances, enzymes and possibly other substances connected with vital processes
Kollath (1953)
Probiotics are opposite of antibiotics
Vergin (1954)
Deleterious effects of antibiotics can be prevented byprobiotics therapy
Kolb (1955)
A substance screted by one microorganism which stimulates the growth of another
Lilly and Stillwell (1965)
Tissue extracts, which stimulate microbial growth
Sperti (1971)
Compounds that build resistance to infection in the host but do not inhibit the growth of microorganisms in vitro
Fuji and Cook (1973)
Organisms and substances that contribute to intestinal microbial balance
Parker (1974)
Live microbial feed supplement which beneficially the host animal by improving microbial balance
Fuller (1992)
Viable mono- or mixed culture of live microorganism which, applied to animal or man, have a beneficial effect on the host by improving the properties of the indigenous microflora
Havenaar and Huisint’Veld (1992)
Live microbial culture or cultered dairy product which beneficially influences the health and nutrition of the host
Salminen et al (1996)
Live microorganism which, upon ingestion in certain numbers, exert health benefits beyond inherent basic nutrition
Schaafsma (1996)
Microbial cell preparations or components of microbial cells that have a beneficial effect on the health and wel-being of the host
Salminen et al (1999)
A preparation of or a product containing viable, defined microorganism in sufficient number, which alter the microflora (by implantation or colonization) in a compatment of the host and by that exert beneficial health effect in this host
Schrezenmeir and de Vrese (2001)
Live microorganisms that when administered in adequate amount confer a health benefit on the host
FAO/WHO (2002)
Azizpour, et al, 2009

5. “Probiotics” is derived from Greek and means “prolife.”
Lilly and Stillwell in 1965: “Growth promoting factors produced by microorganisms.”
Parker in 1974 : “Organisms and substances with beneficial effects for animals by influencing the intestinal microflora.”
Fuller in 1989: “A live microbial feed supplement which beneficially affects the host animal by improving its intestinal microbial balance.”
Havenaar and Huis Int Veld in 1992: “A mono- or mixed culture of live microorganisms which, applied to animal or man, affect beneficially the host by improving the properties of the indigenous microflora.”
ILSI (International Life Sciences Institute) EuropeWorking Group (1998): “A viable microbial food supplement which beneficially influences the health of the host.”
Diplock et al. in 1999 puts it as “Probiotic food is functional if they have been satisfactorily demonstrated to
beneficially affect one or more target functions in the body beyond adequate nutritional effects, in a way that is relevant to either an improved state of health and well-being and/or reduction in the risk of diseases.”
Naidu et al. in 1999: “A microbial dietary adjuvant that beneficially affects the host physiology by modulating mucosal and systemic immunity, as well as improving nutritional and microbial balance in the intestinal tract.”
Tannock in 2000: “Microbial cells which transit the GI tract and which, in doing so, benefit the health of consumer.”
Schrezenmeir and de Vrese in 2001: “A preparation of a product containing viable, defined microorganisms in sufficient numbers, which alter the microflora (by implantation or colonization) in a compartment of the host and by that exert beneficial health effects in this host.”
FAO/WHO (2001) and Reid et al.(2003): “Live microorganisms which when administered in adequate amounts confer a health benefit on the host.”
Kun-Lee and Salminen, 2009

6. Definitions (Harish, 2006)
Probiotic: A live food ingredient which is beneficial to health.
Prebiotic: A non-digestible food ingredient which beneficially affects the host by selectively stimulating the growth and/or activity of one or a limited number of bacteria in the colon having the potential to improve host health.
Synbiotic: A mixture of probiotics and prebiotics which beneficially affects the host by improving the survival and implantation of live microbial dietary supplements in the gastrointestinal tract, and thus improving host health and wellbeing.

7. Present
Kun-Lee and Salminen, 2009

8. FAO/WHO (2002), Guidelines for the Evaluation of Probiotics in Food
FAO/WHO (2002), Guidelines for the Evaluation of Probiotics in Food . Joint Working Group Report on Drafting London, Ontario, Canada, April 30 and May 1, 2002

9. Evaluation
Probiotics have been tested for an impact on a variety of clinical conditions (see Expert Consultation Report, Section 5.3). Standard methods for clinical evaluations are comprized of
Phase 1 (safety),
Phase 2 (efficacy),
Phase 3 (effectiveness) and
Phase 4 (surveillance).
Phase 1 studies focused on safety are discussed in Section 3.3 above.
Phase 2 studies, generally in the form of randomized, double blind, placebo-controlled
(DBPC) design, measure efficacy compared with placebo.
In addition,
phase 2 studies measure adverse effects. A general recommendation for the testing of probiotic foods is that the placebo would be comprized of the food carrier devoid of the test probiotic. Sample size needs to be calculated for specific endpoints. Statistically significant differences must apply to biologically relevant outcomes.
FAO/WHO Report of a Joint Working Gruoup on Drafting Guidelines for the Evaluation of Probiotics in Food. London, Ontario, Canada, 30 April-1 May 2002

10. IDENTIFY THE EXACT STRAIN THAT IS BEING USED TWO
GUIDELINES FOR THE EVALUATION OF PROBIOTICS (Reid,2005)
STEPS
PROCEDURE
ONE
IDENTIFY THE EXACT STRAIN THAT IS BEING USED
TWO
INVITRO EVIDENCE IS INSUFFICIENT TO CALL A STRAIN A PROBIOTICS
THREE
SAFETY OF PROBIOTICS
FOUR
EFFICACY - PROVING THAT THE PROBIOTIC CONFERS A SIGNIFICANT IMPROVEMENT IN HEALTH
FIVE
EFFECTIVENESS
SIX
HEALTH CLAIMS AND LABELING

11. 1. Human origin
2. GRAS status
3. History of safe use in food
4. Documented health benefits
5. Antimutagenic and anticarcinogenic properties
6. Nonpathogenic
7. Tolerance to antimicrobial substances yet inability to tolerance other bacteria
8. Adherence to intestinal mucosa
9. Ability to reduce pathogen adhesion to surfaces
10. Antimicrobial activity against potentially pathogenic bacteria
11.- Immunostimulation without proinflammatory effect
12. Acid tolerance
13. Human gastric juice tolerance
14. Bile toletrance
15. Phage resistance
16. Oxygen and heat tolerance
17. Desired metabolic activity
18. Ability to grow in milk
19. Good sensory properties
20. Retain viability and stability during foor processing, storage, and following consumption
List of Desirable Properties of Probiotic Microorganism for Incorporation into Fermented Food Products Farnworth, 2008

12. Interaction between probiotic, prebiotic and intestinal flora (Harish, 2006)

13. Fermented milks:
dairy products are those obtained by fermentation of milk, which may have been prepared from products obtained from milk with or without changes in the composition. (CODEXSTAN ).
Fermentation of milk by the action of specific microorganisms resulting in the reduction of pH, added or not food additives and optional ingredients (NOM-185-SSA1-2002)

14. Composition of fermented milks
(section 3.3 composition) (CODEXSTAN ).

15. El yogurt:
fermented dairy products.
Symbiotic blend of Streptococcus thermophyllus y Lactobacillus delbrueckii subp. Bulgaricus.
pH less than 4.4
Acidity greater than 0.5% of lactic acid
Probiotic

16. Future
The three focus
Health.
Nutrition ⇋ Food.
Biotechnology

17. Increment the knowledge of the role microorganisms play in human nutrition, immune function and disease resistance increases.
The number of fermented products on the market.
In some products it will be necessary to ensure that the microorganisms consumed are alive
Other products, metabolites and/or fermentation products may be the active ingredient.
Fermented foods will be available for specific niche consumers.
Fermented foods have been part of the human diet for centuries and it may well be that they will become important in the diets of future space travelers.
Farnworth, 2008

18. FUTURE Consumers are increasingly managing their health by self-medication, generating expanding market opportunities for the food and pharmaceutical industry. Food and drink manufacturers are adopting genomic and proteomic technologies previously the domain of the pharmaceutical industry to design more sophisticated and novel products. Because exploitation of the probiotic concept is still associated with unsubstantiated claims reliable and proven products need to be readily distinguished from those of dubious quality. A significant proportion of consumers mistrust manufacturers’ claims. Forty-five percent of Americans claim to largely or entirely disbelieve food and drink manufacturers’ health claims, a figure similar to France and greater than in the Netherlands (538). The evolution of probiotic products thus necessitates changes in the regulations related to labeling, safety, and health claims. The credibilityofhealthclaimsforhealthyindividualsremainstobeestablished.Viable probiotic bacteria have to be consumedin large quantities for a prolonged period to achieve a health benefit. The long-term effects of probiotic consumption on a healthy population are unknown, and yet the general population is being encouraged to consume probiotics regularly, to promote good health and well-being. Studies have yet to be undertaken to demonstrate what effect, if any, there is on the well-being of healthy individuals. While probiotic cultures are incorporated into foods or dietary supplements without making specific health claims they avoid the need to conform to the more rigorous regulatory procedures for therapeutic products, which require demonstrated quality, safety and efficacy. There is evidence from well-conducted clinical trials of beneficial health effects from probiotics in a range of clinical conditions.The probiotic effects have been shown to be strain specific so health effects cannot be generalized between strains. Standardized, verifiable clinical studies are needed to demonstrate the safety, efficacy, and limitations of a putative probiotic, and whether it is superior to existing therapies. Additional studies are needed to determine effects on the immune system in healthy and diseased individuals and effects of long-term consumption. More rigorous quality control, standards, and regulations have been called for (481, 482, 477, 483). The prospect of GM probiotics targeted for clinical conditions demands a rigorous safety strategy to prevent spread into the environment and dissemination of the genetic modification. Permission to label a product probiotic should remain contingent on its compliance with the FAO/WHO definition of probiotic (475). Labels should include consumer or prescriber information about the identity of the organism(s); itsGMOstatus; viability count and shelf life; dosing and duration; conditions for which its use is and is not appropriate; proven benefits; side effects, particularly symptoms that require clinical assessment; and a recommendation to advise health practitioners of probiotic use (471). Where an adverse reaction is suspected the facility to report it along with the 94 PROBIOTIC MICROORGANISMS product details to a national database should be available, as it is for adverse effects of other therapies. Guidelines have been proposed to assess the efficacy and safety of probiotics but international agreement on these has yet to be arrived at. Consensus on uniform regulations is desirable to ensure unequivocal identity, quality manufacturing processes, accurate labeling, proven safety and efficacy for a product that will then merit the label “probiotic.” Kun-Lee and Salminen, 2009

19. 4. Promotion of these guidelines at an international level.
Recommendations
1. Adoption of the definition of probiotics as ‘Live microorganisms which when administered in adequate amounts confer a health benefit on the host’.
2. Use and adoption of the guidelines in this report should be a prerequisite for calling a bacterial strain ‘probiotic’.
3. Regulatory framework to allow specific health claims on probiotic food labels, in cases where scientific evidence exists, as per the guidelines set forth in this report.
4. Promotion of these guidelines at an international level.
5. Good manufacturing practices (GMP) must be applied in the manufacture of probiotic foods with quality assurance, and shelf-life conditions established.
6. Further development of methods (in vitro and in vivo) to evaluate the functionality and safety of probiotics.
FAO/WHO Report of a Joint Working Gruoup on Drafting Guidelines for the Evaluation of Probiotics in Food. London, Ontario, Canada, 30 April-1 May 2002

20. FUTURE PERSPECTIVES
The capacity to assess the gut microbiota has expanded dramatically with the advent of molecular techniques. Real-time quantitative polymerase chain reaction procedures are among the promising tools for studies on intestinal microbiota composition.
Such advancement will lead to the development of a new generation of probiotics, the action of which could be selected for defined disease-associated deviations in gut microbiota. This may also facilitate the potential use of genetically modified probiotic bacteria for pharmaceutical uses. Genetically modified lactic acid bacteria have been proposed as a vehicle to deliver vaccines in the gastro-intestinal tract. Several secretion-expression probiotic vectors have been constructed and are currently being tested in animal models. Other probióticos carrying different immunomodulating molecules are currently being tested. Also, the probiotic vector have been modified to provide a way to deliver the active ingredient at specific targets in the gastrointestinal tract.
Various processing advances, such as microencapsulation and bacterial coating and addition of prebiotic compounds used as growth factors for probiotic organisms, will provide a means to optimize the delivery and survival of strains at the site of action.

21. 1. implementation of Guidelines for the use of probiotics;
THE FUTURE
In summary, the following is a list of clinically relevant steps that will move probiotics closer to being embraced by the medical community:
1. implementation of Guidelines for the use of probiotics;
2. Phase I and II clinical trial data on strains and end products to prove health benefits;
3. Use of Good Manufacturing Practices and production of high quality products;
4. Studies which identify mechanisms of action of probiotic strains in vivo;
5. Appropriate information dissemination about products to physicians, health professionals and lay people;
6. Development of probiotic organisms that carry vaccines or other beneficial substances to the host; vii) development of anti-viral probiotics;
7. Expansion of proven strains to benefit the oral cavity, nasopharynx, respiratory tract, stomach, vagina, bladder, and skin as well as for cancer, allergies and recovery from surgery and injury.
Reid, G The Importance of guidelines in the development and application of probiotics. Current Pharmaceutical Design. 11:11-16

22. Future Implications
There is considerable potential for the benefits of probiotics consumption over a wide range of clinical conditions. On-going research will continue to identify and characterize existing strains of probiotics, identify strain-specific outcomes, and determine optimal doses needed for certain results. According to Barry Goldin, Ph.D., professor at Tufts University School of Medicine, “The full potential of probiotics can only be realized when their benefits can be established scientifically. It is highly likely that benefits from current and future probiotics have gone undetected and, therefore, full utilization of these organisms has not been achieved.”
Daniel O’Sullivan, Ph.D, assistant professor in the Department of Food Science and Nutrition at the University of Minnesota, says regarding yogurt consumption, “At best, your intestinal health is greatly improved and the immune system is strengthened. At worst, there are no adverse effects and you get some nutrients in the bargain.”
With the current focus on disease prevention and the quest for optimal health at all ages, the probiotic market potential is enormous. Health professionals are in an ideal position to help guide their clients toward appropriate prophylactic and therapeutic uses of probiotics that deliver the desired beneficial health effects.
Dairy Council of California 2000, Probiotics – Friendly Bacteria with a Host of Benefits
“Let food be thy medicine and medicine be thy food,” the age-old quote by Hippocrates, is certainly the tenet of today. With the growing interest in self-care and integrative medicine coupled with our healthembracing baby boomer population, recognition of the link between diet and health has never been stronger. As a result, the market for functional foods, or foods that promote health beyond providing basic nutrition, is flourishing. Within the functional foods movement is the small but rapidly expanding arena of probiotics – live microbial food supplements that beneficially affect an individual by improving intestinal microbial balance. The consumers’ overwhelming interest in and demand for functional foods, including probiotics, make it imperative that health professionals stay abreast of the latest research findings and available products. This monograph provides a summary of research on the health benefits of probiotics and offers practical information to help the clinician make appropriate recommendations to clients.

23. Conclusions 1. Implementation of guidelines for the use of probiotics;
2. The clinical trials phase I, II and III to assess the health benefits as a standard for the prevention or treatment for a particular condition or disease;
3. The good manufacturing practices and production of high quality;
4. The studies to identify the mechanism of action in vivo;
5. The accurate labeling information;
6. The development of probiotics that can carry vaccines for the host and / or anti-viral probiotics;
7. The expansion of beneficial strains tested to the oral cavity, nasopharynx, respiratory tract, stomach, vagina, bladder and skin, as well as for cancer, allergies and recovery from surgery / injury

24. REFERENCES:
Azizpour, K., Bahrambeygi, S., Mahmoodpour, S. and Azizpour, A History and Basic of Probiotics. Research Journal of Biological Sciences 4(4):
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25. Mtro. Pedro Javier Guerrero Medina
GRACIAS
Mtro. Pedro Javier Guerrero Medina