Probiotics for Developing Countries

Yuan Kun LEEDepartment of Microbiology,

Yong Loo Lin School of Medicine, National University of Singapore.

5 Science drive 2, Singapore 117597

Intestinal microbiotaBeneficialeffects

Harmfuleffects

Onset of IBD/IBS & others

Infectious disease

Secondary bile acids

Production of harmfulsubstance

Colonization resistance

Production of essential nutrition

Synthesis of vitamins

Immune functions

1000 < bacterial species (1014 < cells)

FAO/WHO

“Probiotics are live microorganisms which when administered in adequate amounts confer a health benefit on the host”

Universal probioticsVsProbiotics for countries of different economic stage of development, dietary habbits & people of different age?

1. Differences in composition of the gut microflora of people (youngsters) from different geographical regions.

Bacteroidaceae

Bifidobacteriaceae

Ruminococcaceae

Lachnospiraceae

Prevotellaceae

Fukuoka

TokyoKorea

Taipei

Taichung

BeijingLanzhou

Bangkok

Khon Kaen

Yogjakarta

Bali

Mongolia

Sichuan

Burkina Faso

Florence

High meat, fat & cereal,Low vegetable

Low meat, fatHigh vegetable & cereal

High meat, fat & cereal, High vegetable

USA

Bangladesh

J Nakayama, K Watanabe, JH Jiang, K Matsuda, SH Chao, P Haryono, O La-ongkham, MA Sarwoko, IN Sujaya, L Zhao, KT Chen, YP Chen, HH Chiu, T Hidaka, NX Huang, C Kiyohara, T Kurakawa, N Sakamoto, K Sonomoto, K Tashiro, H Tsuji, MJ Chen, V Leelavatcharamas, CC Liao, S Nitisinprasert, ES Rahayu, FZ Ren, YC Tsai, YK Lee (2015) Diversity in gut bacterial community of school-age children in Asia. Scientific Reports| 5 : 8397 | DOI: 10.1038/srep08397

Size of the circle correspond to the abundanceRed line indicate negative correlationBlue lines indicate positive correlation

Clostridium difficile colitisIBD

Graft vs host disease

Appendicitis, IBD

Ulcerative colitis, autism

Periodontitis, periapical infection

Peridontitis, Salmonella thyphimurium

Pneumonia, necrotizing fasciitis

Crohn’s disease

Erysipeloid

Obesity, type 2 diabetes

Enterococcus faecium

Butyric acid producing

Meningitis, pneumonia

Shigellosis

Salmonellosis

Kinetics of probiotic adhesion on mucosal surfaceinvolving adhesion-receptor interaction

Lee, Y K and K.Y. Puong, "Competition for adhesion between

probiotics and human gastrointestinal pathogens in the presence of

carbohydrate". BRITISH JOURNAL OF NUTRITION, 88 (2002):

101‑108. Lee, Y K, CY Lim, WL Teng, AC Ouwehand, FM Tuomola and S

Salminen, "Quantitative approach in the study of adhesion of lactic

acid bacteria to intestinal cells and their competition with

enterobacteria". Applied and Environmental Microbiology, 66

(2000): 3692‑3697.

A Mixed probiotics

0.3

0.4

0.5

0.6

0.7

0.8

6 7 8 9 10 11 12

Log Dose/day

1-m

ean

Ris

k R

atio

Borgia et al. 1982; Correa et al. 2005; Gotz et al. 1979; Nista et al. 2004

B Single probiotic

0.4

0.5

0.6

0.7

0.8

0.9

9.4 9.6 9.8 10 10.2 10.4 10.6 10.8

Log Dose/day

1-m

ean

Rish

Rat

io

L. rhamnosus GG

S. boulardii

Adam et al, 1977; Armuzzi zi et al. 2001; Arvola et al. 1999; Correa et al. 2005; Gotz et al. 1979; McFarland et al. 2000; Seki et al. 2003; Surawicz et al. 1989; Tankanow et al;. 1990; Thomas et al. 2001

Prevention of antibiotic-associated diarrhea

Sugars are involved intestinal adhesion of microbes:  Adhesion of microbes on gastrointestinal surface a major prerequisite for their colonization and modulation of local and systemic physiological (immunological) activities.

Lee YK, Puong KY. Competition for adhesion between probiotics & human gastrointestinal pathogens in the presence of carbohydrate. British Journal of Nutrition, 2002, 88 Suppl. 1:101-108.

Lee YK, Puong KY (2002) Competition for adhesion between probiotics and human gastrointestinal pathogens in the presence of carbohydrate. Br J Nutr 88: S101-108

Lee YK, Puong KY (2002) Competition for adhesion between probiotics and human gastrointestinal pathogens in the presence of carbohydrate. Br J Nutr 88: S101-108

Lee YK, Puong KY. Competition for adhesion between probiotics & human gastrointestinal pathogens in the presence of carbohydrate. British Journal of Nutrition, 2002, 88 Suppl. 1:101-108.

Children #220 Adult #150 Senior #128

Gut bacterial composition of children, adult, and senior, living in different Asian regions

Korea Mongolia Korea

China

Thailand

Indonesia

Mongolia Korea

China

Thailand

Indonesia

Bifidobacteriaceae Other Actinobacteria Bacteroidaceae Prevotellaceae Other Bacteroidetes

Ruminococcaceae Lachnospiraceae Streptococcaceae Peptostreptocaccaceae Other Firmicutes

Veillonellaceae Enterobacteriaceae Verucomicobiaceae Fusobacteriaceae Erysipelotrichaceae

Mongolia

China

Indonesia

Thailand

Physiology, lifestyle, use of medication, different probiotics for restoration of a healthy microbiome?

p = 1.69 x 10-13

100

10

1

0.1

0.01

0

(%)100

10

1

0.1

0.01

0

(%)0.00061

0.0021 0.00057 2.0e-08

5.1e-05

5.5e-05

0.013

China Indonesia Korea Mongolia ThailandALL C

hildre

nAdultSe

nior

Child

ren

AdultSe

nior

Child

ren

AdultSe

nior

Child

ren

AdultSe

nior

Child

ren

AdultSe

nior

Child

ren

AdultSe

nior

Change in Bifidobacteriaceae population during aging

p = 1.29 x 10-8

100

10

1

0.1

0.01

0

(%)100

10

1

0.1

0.01

0

(%)0.0077

3.4e-09

1.9e-05 0.011

0.033

0.033

0.016

China Indonesia Korea Mongolia ThailandALL

Change in Enterobacteriaceae population during aging C

hildre

nAdultSe

nior

Child

ren

AdultSe

nior

Child

ren

AdultSe

nior

Child

ren

AdultSe

nior

Child

ren

AdultSe

nior

Child

ren

AdultSe

nior

2. Specific nutritional and health requirements in different geographic regions

1. Probiotic alleviation of lactose intolerance is more relevant for lactose-intolerant Asian people compared to lactose-tolerant European people.

2. Delivery of vitamins, e.g. folic acid (vitamin B11), riboflavin (vitamin B2), cobalamin (vitamin B12), and breakdown of non-nutritive factors like phytic acid that inhibits the absorption of iron, are more relevant for people in malnutrition developing countries and elderly.

3. Prevention of diarrhea or necrotizing enterocolitis, which occurs more frequently in Africa and Asia, among children and preterm infants.

4. Probiotic binding of toxins (heavy metals, aflatoxins) and detoxification can counteract the negative consequences of toxin contamination in foods and feed in developing countries.

3. Survivability in local food matrices

Foods and beverages from Asia, Africa or Southern America are often characterized by presence of herbs and spices with antibacterial properties, such as Capsicum annuum (red pepper), Allium sativum (garlic) and tea.

Probiotics that are more resistant to certain spices and herbs may have an advantage over other probiotics in terms of stability, viability and ultimately functionality after consumption.

Z. M. ROSS, E. A. O’GARA, D. J. HILL, H. V. SLEIGHTHOLME, D. J MASLIN. Antimicrobial Properties of Garlic Oil against Human

Enteric Bacteria: Evaluation of Methodologies and Comparisons with Garlic Oil Sulfides and Garlic Powder. APPLIED AND ENVIRONMENTAL MICROBIOLOGY. 2001, p. 475–480 Vol. 67, No. 1

Lee HC, Jenner AM, Low CS, Lee YK (2006) Effect of tea phenolics & their aromatic fecal bacterial metabolites on intestinal microbiota. Research in Microbiology 159: 876-884.

4. Probiotics are different in their functions

DC cytokine profile induced by exposure to Bifidobacterium strains. Immature DCs were cultured with 12 bifidobacterial strains, E. coli, L. lactis or LPS (bacteria:cell ratio 5:1) and the supernatants were recollected after 48 h for measuring the production of IL-8, IL-6, IL-1β, IL-10, TNFα and IL-12 by multiplex assay. Independent ratios were calculated from each donor. Data represent median and interquartil range of five independent experiments performed with different blood donors. Differences among different bacterial strains were evaluated by Kendall W test: IL-8,Lopez et al., 2010. Distinct Bifidobacterium strains drive different immune responses in vitro. Intl J Food Microbiol 138: 157-165.

5. Technological developments

Growing insights on the mechanism of action of probiotics, metagenome analyses and development of validated biomarkers, e.g. for the production of vitamins, short chain fatty acids, possession of enzymatic activities like phytase or lactase, antimicrobial properties towards locally existing undesired microorganisms, anti-inflammatory profile, H2O2 production, binding to mucus, or ability to degrade certain carbohydrates from local diets. This knowledge further contribute to set a rationale for selection of probiotics.

Regionalized/ personalized probiotics

Wilbert Sybesma, Remco Kort and Yuan-Kun Lee. Locally sourced probiotics, the next opportunity for developing countries? Trends in Biotechnology. 2015, in press.

Differences in composition of the gut microflora

Valorization of ownership of locally sourced probiotics

Technological developments

Specific nutritional and health requirements

Survivability of probiotics in local food matrices

Thank you!