November 16, 2014 1Prof. Drh. Hj. ENDANG PURWATI R N, MS.,

Ph.D

International Conference on Advances in Healthcare & Life Sciences

(Singapore) 8 – 9 November 2014

INFLUENCE OF Weisella paramesenteroides ISOLATE “DADIH”TO THE BOWEL FREQUENCE, SECRETORY

IMMUNOGLOBULIN A LEVEL AND HEIGHT OF ILEUM VILLI OF THE MICE EPEC INDUCED DIARRHEA

November 16, 2014 2Prof. Drh. Hj. ENDANG PURWATI R N, MS.,

Ph.D

ANDALAS UNIVERSITYPADANG, WEST SUMATERAINDONESIA

Laboratory of Biotechnology / Technology of Product Husbandry, Faculty of Animal Science, Andalas University, Padang, West Sumatera,

IndonesiaEmail : purwati17@yahoo.co .id

INTRODUCTION

November 16, 2014Prof. Drh. Hj. ENDANG PURWATI R N, MS.,

Ph.D3

Benefit of Research

researchers interested in studying the effect of these bacteria on diarrhea particularly levels of SigA and ileum villi.

November 16, 2014Prof. Drh. Hj. ENDANG PURWATI R N, MS.,

Ph.D10

MATERIAL AND

METODERESEARCH

November 16, 2014Prof. Drh. Hj. ENDANG PURWATI R N, MS.,

Ph.D11

• Experimental animals were randomized into two factors:

• The first factor (A) is the dose of probiotic administration, consist of a negative control: only given standard feed and water (A1), positive control: EPEC administered at a dose of 108 cfu/g (A2), W. paramesenteroides dose of 2 x 108cfu/g (A3), W. paramesenteroides dose of 2 x 109cfu/g (A4), W. paramesenteroides with a dose of 2 x 1010cfu/g.

• The second factor (B) is the duration of observation (hour), which consist of 0 hour (B1), 12 hours (B2), 24 hours (B3), 36 hours (B4)

Observed variables :

1. Examination of bowel frequency, 2. levels of SIgA and 3. height of ileum villi

November 16, 2014Prof. Drh. Hj. ENDANG PURWATI R N, MS.,

Ph.D14

November 16, 2014Prof. Drh. Hj. ENDANG PURWATI R N, MS.,

Ph.D15

RESULTS AND DISCUSSION RESULTS AND DISCUSSION

Tabel 1. Mean bowel frequency(times) of mice based on doses and duration of observation

November 16, 2014Prof. Drh. Hj. ENDANG PURWATI R N, MS.,

Ph.D16

A Factor(doses)

B Factor (duration of observation)

Total Mean SDB1= 0hour

B2=12 hours

B3 = 24 Hours

B4 = 36hours

A1=K (-) 4,00g 5,00g 4,50g 5,00g 18,50 4,63 1.52 A2=K (+)

5,00g 50,50a 41,00b 37,00c 133,50 33,384.09

A3=2x108

5,50g 48,50a 16,50e 11,00f 81,50 20,38

3.19

A4=2x109

5,00g 30,50d 20,00e 16,00e 71,50 17,882.99

A5=2x1010

5,50g 28,50d 12,50f 11,00f 57,50 14,382.68

Total 25,00 163,00 94,50 80,00 362,50 90,63Mean 5,00 32,60 18,90 16,00 72,50 18,13

Note : different small letter superscripted in different columns and rows has highly significant difference (P<0.01).

Table 2. MeanSIgA (µg/ml) in feces based on doses and duration of observation

November 16, 2014Prof. Drh. Hj. ENDANG PURWATI R N, MS.,

Ph.D17

Factor A(dose)

Factor B (duration of observation)

Total Mean SDB1 = 0 hours

B2 = 12 hours

B3 = 24 hours

B4 = 36 hours

A1 = K(-) 1.82e 1.80e 1.82e 1.81e 7.26 1.82 0.95A2 = K(+) 1.83e 1.57h 1.60g 1.66f 6.66 1.66 0.91A3 = 2x108 1.84e 1.53h 2.27b 2.63a 8.59 2.15 1.04A4 = 2x109 1.83e 1.59h 2.13c 2.26b 7.80 1.95 0.99A5 = 2x1010

1.82e 1.56h 1.96d 2.17c 7.52 1.880.97

Total 9.14 8.05 10.24 10.40 37.83 9.46

Mean 1.83 1.61 2.05 2.08 7.57 1.89

Note : different small letter superscripted in different columns and rows has highly significant difference (P<0.01).

Table 4. Mean of height of ileum villi (µm) based on doses and duration of observation

November 16, 2014Prof. Drh. Hj. ENDANG PURWATI R N, MS.,

Ph.D18

Factor A(Dose)

Factor B (duration of observation)

Total Mean SDB1 = 0 hours

B2 = 12 hours B3 = 24 hours B4 = 36 hours

A1 = K(-) 111.21g 111.01g 115.49g 116.66g 454.37 113.59 7.54

A2 = K(+) 111.86g 54.33k 91.38h 116.73g 374.30 93.58 6.84

A3 = 2x108 111.17g 66.72j 156.42d 187.26a 477.10 119.28 8.07

A4 = 2x109 112.32g 72.68i 129.19f 164.06c 535.72 133.93 7.73

A5 = 2x1010 114.99g 52.08k 150.54e 174.80b 482.41 120.60 7.85

Total 564.55 356.82 643.02 759.51 2323.90 580.98

Mean 112.91 71.36 128.60 151.90 464.78 116.20

Note : different small letter superscripted at different columns and rows showed highly different significance (P<0.01).

Conclusion

Giving W.paramesenteroides dosing 2x108cfu/g after 24 hours significantly decreased the frequency of bowel, increased levels of SIgA and improving ileum villi in mice EPEC-induced diarrhea.

November 16, 2014Prof. Drh. Hj. ENDANG PURWATI R N, MS.,

Ph.D19

Acknowledgements

This research was supported by Laboratory of Biotechnology / Technology of Product Husbandry, Faculty of Animal Science, Andalas University, Padang. Indonesia and Hibah Pascasarjana Nomor : 004/UN.16/PL/MT-HB-PC/2012 Date 24 January 2012 Prof. Drh. Hj. Endang Purwati, MS, Ph.D.

November 16, 2014Prof. Drh. Hj. ENDANG PURWATI R N, MS.,

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Anggarini W, Muchtadi D, Astawan M, Wresdiati T. Effect of indigenous probiotic in sinbiotic yoghurt toward histological profile and imunoglobulin A content in small intetine of mice (Tesis). Bogor: Bogor Agricultural University; 2011.

Astawan M , Wresdiyatib T, Ariefc II, Suhestia E. Gambaran hematologi tikus putih (rattus norvegicus) yang diinfeksi escherichia coli enteropatogenik dan diberikan probiotik. Med Peternakan 2011:7-13.

Baratawidjaya KG. Imunologi dasar. 7 ed. Jakarta: Balai Penerbit FKUI; 2006.

Branski D, Wilschanski M. Probiotics in gastrointestinal disorder. In: Kliegman RM, Behrman RE, Jenson HB, Stanton BF, eds. Nelson textbook of pediatrics. Philadelphia: Saunders; 2007:1618-20.

Budiño FEL, et al. Effect of Probiotic and Prebiotic Inclusion in Weaned Piglet Diets on Structure and Ultra-structure of Small Intestine. Brazilian archives of Biology and Technology 2005;48:921-30.

Donowitz M, Alan NC, Richard H, Samuel B, Collins H. Significance of abnormal rabbit ileal histology in the pathogenesis of diarrhea. Infection and Immunity 1979;26:380-6

November 16, 2014Prof. Drh. Hj. ENDANG PURWATI R N, MS.,

Ph.D21

Fukushima Y, Kawata Y, Hara H, Terada A, Mitsuoka T. Effect of a probiotic formula on intestinal immunoglobulin A production in healthy children. Int J Food Microbiol 1998;42:39-44.

Galdeano CM, et al. Impact of a probiotic fermented milk in the gut ecosystem and in the systemic immunity using a non-severe protein-energy-malnutrition model in mice. BMC Gastroenterology 2011;11:1-14.

Haq JA, Li HC, Rahman RA. Detection of enteropathogenic escherichia coli (EPEC) by serotyping and cell adhesion assay among children in North Eastern Peninsular Malaysia - a hospital based study. Ibrahim Med Coll J 2008;2:40-3.

Isolauri E, Sutas Y, Kankaanpaa P, Salminen S. Probiotic : effects on immunity. Am J Clin Nutr 2001;73:444S-50S.Liu J, et al. Effect of Lianshu preparation on lipopolysaccharide-induced diarrhea in rats. World J Gastroenterol :

2009 15(16):2009-15.Lodinová-Zádníková R, et al. The antibody response in breast-fed and non-breast-fed infants after artificial

colonization of the intestine with Escherichia coli 083. Ped Res 1991;29:396-9.Mair C, et al. Inulin and probiotics in newly weaned piglets: effects on intestinal morphology, mRNA expression

levels of inflammatory marker genes and haematology. Arch Anim Nutr 2010;64(4):304-21.Minelli EB, Benini A. Relationship between number of bacteria and their probiotic effects. Microbial Ecology in

Health and Disease 2008;20:1803.Papagianni M, Papamichael EM. Purification, amino acid sequence and characterization of the class IIa bacteriocin

weissellin A, produced by Weissella paramesenteroides DX. Bioresource Technology 2011;102:6730-4.Paul J, Verma AK, Verma R. Role Of Gut Flora In Inflammatory Bowel Disease- A State Of Art. In: Communicating

Current Research and Educational Topics and Trends in Applied Microbiology. New Delhi; 2007:705-15.Pelicano ERL, et al. Intestinal mucosa development in broiler chickens fed natural growth promoters. Brazilian

Journal of Poultry Science 2005;7:221-9.

November 16, 2014Prof. Drh. Hj. ENDANG PURWATI R N, MS.,

Ph.D22

Prasetyo DH, Purwanto B. Efek Probiotik pada Kadar IgA Mencit Model Sepsis. MKB 2010;42:175-80.Purwati, E., Arief dan Ade, R. 2011. Teknologi Dadiah. Cendekia, Bogor. ISBN 978-979-15949-8-1.Reid G, Jass J, Sebulsky T, McCormick JK. Potential Uses of Probiotics in Clinical Practice. Clinical Microbiology

Review 2003:658-72.Retnaningtyas LP, Sudarmo SM, Harsono A, Damanik SM. Effect of probiotic on the fecal sIgA level in preterm

infants (A randomized double-blind placebo control study). Paediatr Indones 2008;48:225-30.Saavedra JM. Use of probiotic in pediatrics: rationale, mechanism of action, and practical aspects. Nutr Clin Pract

2007;22:351-65.Samanya M, Yamauchi K. Histological alterations of intestinal villi in chickens fed dried Bacillus subtilis var. natto.

Comparative Biochemistry and Physiology 2002;133:95-104.Shobha RP, Agrawal R. Volatile Compounds of Therapeutic Importance Produced by Leuconostoc

paramesenteroides, a Native Laboratory Isolate. Turk J Biol 2007;31 35-40.Steel RGD, Torrie JH. Principles and procedures of statistics a biometrical approach. 2 ed. Singapore: McGrow-Hill

Book Campany; 1986.Strugnell RA, Wijburg OLC. The role of secretory antibodies in infection immunity. Nature 2010;8:656-65.Subagyo B, Santoso NB. Diare Akut. In: Juffrie M, Soenarto SSY, Oswari H, Arief S, Rosalina I, Mulyani NS, eds. Buku

ajar gastroenterologi hepatologi. 1 ed. Jakarta: Badan Penerbit IDAI; 2010:87-120.Sulistiyani I. Gambaran mikroskopis permukaan vili usus halus ayam broiler setelah pemberian probiotik dan

bioinsektisida (Tesis). Bogor: Institut Pertanian Bogor; 2003.Taheri HR, et al. Efficacy of Pediococcus acidlactici-based probiotic on intestinal Coliforms and villus height, serum

cholesterol level and performance of broiler chickens. African Journal of Biotechnology 2010;9:7564-7.

Trabulsi LR, Keller R, Gomes TAT. Typical and Atypical Enteropathogenic Escherichia coli. Emerging Infectious Diseases 2002;8:508 - 13.

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THANK YOU

November 16, 2014Prof. Drh. Hj. ENDANG PURWATI R N, MS.,

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