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5/26/2009

Inggrid Surono

Inggrid Surono

5/26/2009

Inggrid Surono

Function of Indigenous probiotic from dadih Enterococcus faecium IS-27526 in improving nutritional status and suppress diarrhoea

Ingrid S Surono*, Fiona Koestomo, F Riewpassa, Yulianasari and Netty Harnetty


*SEAMEO TROPMED RCCN University of Indonesia Salemba Raya 6, Jakarta 10430 INDONESIA

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Dadih, Indonesian traditional fermented raw buffalo milk of West Sumatra

Enterococcus faecium IS IS-27526 strain isolated from Dadih possess probiotic properties

Probiotic Properties

high viability acid and bile tolerance


adhesion

colonization Immuno-modulatory Antagonisms against pathogens antimutagenic, hypocholesterolemic Safety in food, clinical safety processing, shelf life, probiotics stability

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TRADITIONAL WAY OF DADIH MAKING

Fresh raw unheated buffalo milk, poured into bamboo tubes

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Diarrhoea in a year globally


4000 million
diarrhaea cases

1500 million
episode of diare children younger than 5

1.8 million die

Every 3 seconds 1 child dies from diarrhoea 70% cases related to contaminated food and drink

Cause of mortality of Indonesian children


INFANTS
Nerve system problem 3% Tetanus 3% Diarrhea 9% Digestive tract problem 4% ARI 28%

UNDER FIVE CHILDREN

Other 17% Perinatal 36%

Other 35%

ARI 23%

Diarrhea 13% Digestive tract problem 6% Nerve system problem 12%

Typhoid 11%

IDHS 2002-2003, HHS 2001

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Human study in Cianjur Public Health Center


Randomised Placebo Control Trial, Pre-post study Three groups n = 35 subjects, 90 days supplementation Probiotic E faecium IS-27526 108 cfu/day

Effect of supplementation fish protein concentrate biscuit with probiotic cream on response immune humoral of children younger than five
1. Imunoglobulin A (IgA) IgA of three groups before and after intervention

Treatment Before Normal biscuit Fish Protein Concentrate (FPC) Biscuit FPC + Probiotic cream 87.79 98.3 86.78 15.28 87.79 33.92

Serum Ig A After 91.74 12.04 95.01 17.94 107.61 38.21 Delta 3.95 3.28* 8.23 4.62** 19.83 16.35**

* significant (P<0.05) ** very significant (P<0.01)

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Fecal Microbiota of children younger than five, before and after supplementation with FPC biscuit with probiotic cream

Profile of fecal microbiota 1 2 3 4 5 Fecal Aerobic bacteria (cfu/g) Log cfu/g Fecal Coliform (cfu/g) Log cfu/g Fecal lactic bacteria (cfu/g) Log cfu/g Fecal Anaerobic bacteria (cfu/g) Log cfu/g Fecal lactic Anaerobic (cfu/g) Log cfu/g

before 7.10 0.96 1.2 x 107 5.40 0.87 2.5 x 105 4.48 3.19 3.0 x 104 6.38 0.92 2.5 x 106 5.61 0.71 4.0 x 105

after 6.42 1.36 2.5 x 106 5.50 0.98 3.1 x 105 6.35 0.70 2.2 x 106 6.29 1.23 1.9 x 106 6.23 0.99 1.5 x 106

delta 0.96 1.65 0.07 1.28 1.86 3.41 0.09 1.15 0.62 0.45**

** significantly different before and after (P<0.001)

2. Fekal Mikrobiota pada Feses Balita

Morbidity rate during 90 days supplementation

Number of subjects FPC biscuit + Probiotic cream (n=35) n 4 6 5 0 0 0 15 % 3.8 5.7 4.8 0 0 0 14.3

No.

Normal Biscuit (n=35) n % 5.7 3.8 7.6 2.9 5.7 3.8 29.5

FPC Biscuit (n=32) n 8 2 4 0 4 4 21 % 8.3 2.1 4.2 0 4.2 4.2 23.0

1 2 3 4 5 6 Total

Diarrhoea High Fever Common cold Dermatitis Fever Cough

6 4 8 3 6 4 31

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Morbidity of children younger than five subjects during 90 days intervention study
Treatment Groups
Normal biscuit FPC biscuit FPC + Probiotic cream

Day 0
Diarrhoea High fever Common cold Dermatitis Fever Cough Measle Total 5 1 1 4 5 0 16

Day 90
1 0 3 2 1 1 0 8

Day 90
2 0 0 0 0 0 0 2

30
2 2 2 1 2 1 0 11

60
3 2 3 0 2 2 0 12

0
3 0 4 0 4 6 1 18

30
2 1 2 0 2 2 0 9

60
4 1 2 0 2 2 0 11

0
6 0 3 0 5 6 0 20

30
4 2 3 0 0 0 0 9

60
0 3 1 0 0 0 0 4

90
0 1 1 0 0 0 0 2

The improvement of Intestinal vili induces by probiotic LIS 10506 and LIS 20506 A: Normal Vili ; B: Disruption o of vili caused by LPS E.coli ; C: the improvement vili by LIS 10506 ; D: the improvement vili by LIS 20506

(Reza Ranuh et al., 2008)

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Protein Intestinal's Brush Border


c o n c e n tr a tio n i n p p m 80000 70000 60000 50000 40000 30000 20000 10000 0 I le u m I le u m I le u m J e ju n u m J e ju n u m J e ju n u m D uodenum D uodenum D uodenum D uodenum J e ju n u m I le u m

LPS Placebo LIS10506 LIS20506

Galectin-4

Myosin-1a

Occludin

ZO-1

(Reza Ranuh et al., 2008)

Distribusi Ekspresi Protein Brush Border Usus Expression of intestinal brush border protein
30
p<0,05) p<0,05) p<0,05) p<0,05)

Ting kat E ksp resi

25 20 15 10 5 0 D J G I D

KONTROL control LPS LPS + LIS 10506


P>0,05)

LPS + LIS 20506

J M

J ZO

J O

Proteinbrush Brush Border Usus Intestinal border protein

(Reza Ranuh et al., 2008)

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0 0 -0.5 30 60 90

Z sco re s

-1

-1.5

-1.59

Z skor -1.88
-2.03 -1.97 -2.02

(BB/U)
-2.01 -2.09

-1.79 -1.8

-2

-2.12 -2.23

-2.12

-2.5

Days
Biskuit Normal biscuit Biskuit KPI FPC biscuit Biskuit KPI+Probiotik FPC + probiotic cream

Z scores (weight/age)
Gambar 19

0 0 -0.5 30 60 90

Z scores

-1
-1.54 -1.59

-1.5 -2 -2.5

-1.72

-1.71 -1.77 -1.66 -1.76 -1.83 -1.84

-2.08 -2.35

-2.08

Days
Biskuit normal biscuit Biskuit KPI FPC biscuit Biskuit KPI+Probiotik FPC + probiotic cream

Z scores (height/age)

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0 -0.2 -0.4 0 30
-0.48

60
-0.56

90 -0.16

Z scores

-0.6 -0.8 -1 -1.2


-1.39 -0.75 -0.93 -0.9 -0.82 -1.1 -1.28

-0.61

-0.93

-1.4 -1.6

Days
Biskuit Normal biscuit Biskuit KPI FPC biscuit Biskuit KPI+Probiotik FPC + probiotic cream

Z scores (weight/height)
Gambar 21

Effect of supplementation on nutritional status of children less than five


Before intervention Z scores Normal biscuit (n=35) n Weight/age Normal underweight Height/age Normal Stunted Weight/height Normal Wasted 31 4 88.6 11.4 25 7 78.1 21.9 33 2 91.4 5.8 34 1 97.1 2.9 31 1 96.9 3.1 35 0 100 0 14 21 40 60 18 14 56.2 43.8 14 21 40 60 14 21 40 60 19 13 59.4 40.6 19 16 54.3 45.7 12 23 34.3 65.7 6 26 18.8 81.2 4 31 11.4 88.6 20 15 57.1 42.0 14 18 43.8 56.2 15 20 42.9 57.1 % FPC biscuit (n=32) n % FPC + Probiotic cream (n=35) n % Biskuit (n=35) After intervention Biskuit KPI (n=32) Biskuit KPI + Probiotik (n=35) n %

Note : weight/age: Normal -2<Z score<2 height/age: Normal : Z score -2 underweight -3<Z score<-2 Stunted : Z score < -2

weight/height : Normal : Z score -2 Wasted : Z score < -2

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Human Study in Teluk Naga Public Health Center Tangerang

Randomised Placebo Control Trial, Pre-post study Two groups, n = 40 subjects, 90 days supplementation Probiotic E. faecium IS-27526 108 cfu/day

0.800 Consentration (mg/ml)

Total Serum IgA


0.267 0.231 0.220 0.274

0.400

0.000 Placebo Group 0d 90 d Probiotic

Total serum IgA (mg/ml), before and after intervention


Placebo : maltodextrin Probiotic : 108 cfu/day Enterococcus faecium IS-27526

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The changes of total serum IgA

0.200 0.1 50 0.1 00 0.1 41 0.1 42

(mg/ml)

0.052 0.050 0.004 0.000 no rmal -0.050 -0.1 00 underweight severe underweight -0.054 0.01 9

Nutritional Status (Bodyw e ight/age )


P lacebo P ro bio tic

Total serum IgA levels (mg/ml) based on nutritional status of subjects

3.200 Consentration (mg/ml)

Total Saliva s IgA


2.400 1.600 0.800 0.000 Placebo Gr oup 0d 90 d 0.160

**

**
0.560 0.136

1.055

Probiotic

Total sIgA (mg/ml) before and after intervention


*) significantly different (p<0.05) **) significantly different (p<0.01)

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The increasing of total saliva IgA

1 .200 1 .000

1 .1 09

0.802 0.800

(mg/ml)

0.595 0.600 0.400 0.200 0.000

0.584 0.352 0.352

no rmal

underweight

severe underweight

Nutritional Status (Bodyw e ight/age )


placebo pro bio tic

Total sIgA (mg/ml) before and after intervention, based on the nutritional status of children less than five

11.6

placebo
11.2

probiotic

Body Weight (kg)

10.8 10.4 10.0 9.6 0 1 2 3 4 5 6 7 8 9 10 11 12 13

Weeks

The increase of body weight during 90 days supplementation


*) significantly difference (P <0.05) **) significantly difference (P <0.01) as compare to weeks 0 (before treatment)

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So, which bacteria did you eat today?

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