Penggantian Posisi dan Penghambatan Bacteroides vulgatus Oleh Beberapa Bakteri Probiotik.

ABSTRAK

PENGGANTIAN POSISI DAN PENGHAMBATAN Bacteroides vulgatus OLEH

BEBERAPA BAKTERI PROBIOTIK

Anggraeini Puspitasari, 2007.

Pembimbing I : Philips Onggowidjaja, S.Si.,M.Si. Pembimbing II : Triswaty Winata, dr., M.Kes.

Bakteri probiotik dapat memperbaiki keseimbangan mikrobiota usus dengan cara berkompetisi melalui kemampuan beradhesi, menggantikan dan menghambat adhesi bakteri patogen. Tujuan penelitian ini mengetahui kemampuan beradhesi bakteri probiotik yang digunakan, yaitu; Bifidobacterium Bb12 (Bb12), Lactobacillus rhamnosus GG (LGG), Lactobacillus plantarum IS-10506, dan Lactobacillus plantarum IS-20506 dalam menggantikan posisi dan menghambat bakteri patogen yang digunakan, yaitu

Bacteroides vulgatus.

Penelitian ini bersifat eksperimental laboratorik prospektif dengan menggunakan metode mucus adhesion assay. Bakteri patogen yang telah diradiolabel dibaca dengan menggunakan liquid scintilation counter untuk penghitungan jumlah bakteri patogen yang masih menempel pada mukus.

Hasil penelitian menunjukkan bahwa Lactobacillus rhamnosus GG dan

Lactobacillus plantarum IS-20506 secara signifikan (P<0,05) menggantikan posisi

adhesi Bacteroides vulgatus masing-masing 70,44%; dan 70,56%. Bifidobacterium Bb12, dan Lactobacillus plantarum IS-10506 dalam penelitian ini tidak berbeda signifikan (P>0,05) dalam menggantikan posisi adhesi Bacteroides vulgatus pada mukus. Keempat bakteri probiotik dalam penelitian ini tidak berbeda signifikan (P>0,05) dalam menghambat adhesi Bacteroides vulgatus pada mukus.

Kata kunci : adhesi, penggantian posisi, penghambatan, bakteri probiotik, Bacteroides

vulgatus

Universitas Kristen Maranatha

ABSTRACT

DISPLACEMENT AND INHIBITION OF Bacteroides vulgatus BY PROBIOTIC BACTERIA

Anggraeini Puspitasari, 2007.

Advicer : Philips Onggowidjaja, S.Si., M.Si. Co Advicer : Triswaty Winata, dr., M.Kes.

Probiotic bacteria are thought to improve resistance to gut infections by competing with pathogen through displacement and inhibition the growth of harmful bacteria and balancing the intestinal microbiota. The aim of this research was to find out the ability of the probiotic bacteria; Bifidobacterium Bb12 (Bb12), Lactobacillus rhamnosus GG (LGG), Lactobacillus plantarum IS-10506, and Lactobacillus plantarum IS-20506 in the displacement and inhibition of pathogenic bacteria Bacteroides vulgatus.

Mucus adhesion assay was carried out. Radiolabeled pathogenic bacteria were counted by liquid scintilation counter, expressed as remaining of pathogenic bacteria adhere on mucus.

The results showed that Lactobacillus rhamnosus GG, and Lactobacillus plantarum IS-20506 were significantly (P<0,05) displaced the Bacteroides vulgatus adhesion by 70,44 % and 70,56 %, respectively, there was no significant (p>0,05) difference of Bifidobacterium Bb12 and Lactobacillus plantarum IS-10506 in displaced the Bacteroides vulgatus adhesion, and there was no significant (P>0,05) difference of four tested probiotic bacteria in inhibiting Bacteroides vulgatus adhesion.

Key words: Adhesion, displacement, inhibition, probiotic bacteria, Bacteroides vulgatus

Universitas Kristen Maranatha

Universitas Kristen Maranatha

Universitas Kristen Maranatha

DAFTAR ISI

. Halaman

LEMBAR PERSETUJUAN ...i

SURAT PERNYATAAN ...ii

ABSTRAK ... iii

ABSTRACT ...iv

KATA PENGANTAR ...v

DAFTAR ISI ...vii

DAFTAR TABEL ...x

DAFTAR GAMBAR ...xi

DAFTAR BAGAN ...xii

DAFTAR LAMPIRAN ... xiii

BAB I. PENDAHULUAN 1.1Latar Belakang ...1

1.2Identifikasi Masalah ...2

1.3Maksud dan Tujuan ...2

1.4Kerangka Pemikiran ...2

1.5Hipotesis Penelitian ...3

1.6Metodologi Peneliltian ...3

1.7Lokasi dan Waktu ...3

BAB II. TINJAUAN PUSTAKA 2.1 Saluran Pencernaan ...4

2.1.1 Mukus Traktus Gastrointestinal ...4

2.1.2 Flora Normal Traktus Gastrointestinal ...5

2.1.3 Bakteri Asam Laktat ...6

2.2 Bakteri Probiotik ...7

2.2.1 Bifidobacterium ...10

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2.2.2 Lactobacillus rhamnosus GG ...12

2.2.3 Lactobacillus plantarum IS-10506 dan Lactobacillus plantarum IS- 20506 ...14

2.3 Flora Komensal ...16

2.3.1 Bacteroides vulgatus...16

2.4 Interaksi Bakteri Probiotik dan Bakteri Patogen ...18

2.4.1 Mekanisme Penghambatan Patogen ...19

A. Kompetisi Beradhesi ...19

B. Pertumbuhan didalam Pencernaan ...20

C. Imunomodulasi dan Stimulasi Innate Immunity ...20

D. Senyawa Antimikrobial ...21

E. Produksi Bakteriosin ...21

2.4.2 Peranan Probiotik dalam Kesehatan ...22

2.4.2.1 Gangguan Saluran Pencernaan ...23

2.4.2.1 Resistensi terhadap Antibiotik ...25

BAB III. ALAT, BAHAN DAN CARA KERJA 3.1 Alat dan Bahan ...26

3.2 Cara Kerja ...29

3.2.1 Persiapan Penelitian ...29

3.2.1.1 Pembuatan Larutan Dapar Hepes-Hanks ...29

3.2.1.2 Pembuatan Larutan SDS 1% NaOH 0.1 M ...29

3.2.1.3 Persiapan Mukus ...30

3.2.2 Pembiakan Bakteri ...30

3.2.2.1 Pembiakan Bakteri Patogen ...31

3.2.2.2 Pembiakan Bakteri Probiotik ...31

3.2.3 Langkah Penelitian Mucus Adhesion Assay ...32

3.2.3.1 Pencucian Kultur Bakteri ...32

3.2.3.2 Menstandarisasi Jumlah Bakteri menjadI 107-108 CFU/ml ...32

3.2.3.3 Pencucian Mukus ...33

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3.2.3.4 Penanaman Bakteri pada Mukus ...33

3.2.3.5 Penghitungan Bakteri ...34

3.3 Analisa Data ...35

3.3.1 Hipotesis Statistik ...35

3.3.2 Kriteria Uji ...35

BAB IV. HASIL PENELITIAN DAN PEMBAHASAN 4.1.Hasil Percobaan Penggantian Posisi ...36

4.1.1 Pembahasan ...37

4.1.2 Pengujian Hipotesis Penelitian ...37

4.2. Hasil Percobaan Penghambatan ...39

4.2.1 Pembahasan ...39

4.2.2 Pengujian Hipotesis Penelitian ...40

BAB V. KESIMPULAN DAN SARAN 5.1 Kesimpulan ...41

5.2 Saran ...41

DAFTAR PUSTAKA ...42

LAMPIRAN ...47

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DAFTAR TABEL

DAFTAR TABEL

2.1 Efek bakteri Probiotik untuk Kesehatan ...23 4.1 Data terindeks presentase penggantian posisi Bacteroides vulgatus oleh keempat bakteri Probiotik ...36 4.2 Uji ANAVA Penggantian posisi Bacteroides vulgatus oleh

Lactobacillus rhamnosus GG ...37 4.3 Uji ANAVA Penggantian posisi Bacteroides vulgatus oleh

Bifidobacterium Bb12 ...38 4.4 Uji ANAVA Penggantian posisi Bacteroides vulgatus oleh

Lactobacillus plantarum IS-10506 ...38 4.5 Uji ANAVA Penggantian posisi Bacteroides vulgatus oleh

Lactobacillus plantarum IS-20506 ...38 4.6. Data terindeks penghambatan Bacteroides vulgatus oleh bakteri

Probiotik...39 4.7 Uji ANAVA Penghambatan Bacteroides vulgatus oleh keempat bakteri Probiotik ...40

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DAFTAR GAMBAR

DAFTAR GAMBAR

2.1 Saluran Pencernaan ...4

2.2 Bakteri asam laktat yang beradhesi pada permukaan mukosa usus...5

2.3 Bifidobacterium Bb12...11

2.4 Bifidobacterium. spp ...11

2.5 Lactobacillus rhamnosus GG. ...13

2.6 Lactobacillus. spp ...15

2.7 Bacteroides vulgatus ...17

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DAFTAR BAGAN

DAFTAR BAGAN

3.1 Rangka pekerjaan penelitian ...28

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DAFTAR LAMPIRAN

LAMPIRAN

Data Terindeks Pengamatan Penggantian posisi ...47 Data Terindeks Pengamatan Penghambatan ...48

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LAMPIRAN 1

Data Terindeks Pengamatan Penggantian posisi

Indeks

Vulgatus = 100 data terindeks

Tanggal LGG BB12 Lp IS-10506

Lp IS-20506

Vulgatus

(control) Rata2 Vulgatus Vulgatus (100%) LGG (%) BB12 (%) Lp IS-10506 (%) Lp IS-20506 (%)

06/05/2007 155 113 169 96 88 64

06/05/2007 177 124 146 83 100 70

06/05/2007 120 96 214

176

121 68 54

06/07/2007 112 161 195 445 148 25 53 65

06/07/2007 198 277 163 205 68 44 92 54

06/07/2007 166 244 138 254

301

84 37 81 46

06/08/2007 125 135 150 186 197 117 74 80 89 111

06/08/2007 185 122 137 155 132 79 110 73 82 92

06/08/2007 148 144 196 133 175

168

104 88 86 117 79

06/12/2007 433 172 77 194

06/12/2007 448 259 116 200

06/12/2007 437 240

224

107 195

13/6/2007 340 204 136 227

13/6/2007 357 131 88 239

13/6/2007 305 114

150

76 204

6/15/07 104 111 97 91

6/15/07 78 111 97 68

6/15/07 71 120

114

105 62

AVERAGE 100 70,44 117,22 131 70,56

LAMPIRAN 2

Data Terindeks Pengamatan Penghambatan

Tanggal LGG BB12 Lp IS-10506

Lp

IS-20506 Vulgatus

Rata2

Vulgatus Vulgatus(100%)

LGG (%)

BB12 (%)

Lp IS-10506 (%)

Lp IS-20506 (%)

26-5-07 149 278 194 183 190 89 69 130 90 85

26-5-07 358 319 206 203 221 103 167 149 96 95

26-5-07 222 315 211 167 233

215

109 103 147 98 78

29-5-07 182 184 173 209 151 67 81 81 77 92

29-5-07 188 187 171 233 218 96 83 83 76 103

29-5-07 201 180 163 203 309

226

137 89 80 72 90

30-5-07 109 208 161 109 192 91 52 99 76 52

30-5-07 176 114 245 155 249 118 83 54 116 73

30-5-07 136 145 181 178 192

211

91 64 69 86 84

AVERAGE 100 87,89 99,11 87,44 83,56

RIWAYAT HIDUP

Nama : Anggraeini Puspitasari

Tempat, tanggal lahir : Jakarta, 10 Februari 1985 Alamat : Danau Bogor Raya C2/2, Bogor

Pendidikan

1. 1990 lulus TK Hosana, Bogor 2. 1991-1993 SD Budi Mulia, Bogor

3. 1993-1996 SD Nishiminowa, Nagano JAPAN 4. 1997 lulus SD Budi Mulia, Bogor

5. 2000 lulus SLTP Budi Mulia, Bogor 6. 2004 lulus SMU Regina Pacis, Bogor

BAB I

PENDAHULUAN

1.1Latar belakang

Banyak bakteri “menguntungkan” dan “ merugikan” yang menghuni usus manusia. Untuk mendapatkan pencernaan yang sehat adalah dengan berusaha agar saluran pencernaan dihuni oleh lebih banyak bakteri “menguntungkan” dibanding bakteri “merugikan”.

Lactobacillus dan Bifidobacteria adalah bakteri asam laktat yang

merupakan bakteri probiotik. Keduanya dapat diperoleh dari susu fermentasi yoghurt, mampu melindungi traktus gastrointestinal dari serangan bakteri patogen. Daya tahan tubuh sangat diperlukan oleh manusia untuk menghadapi berbagai penyakit. Manusia memiliki kekebalan tubuh dengan berbagai macam bentuk. Contohnya kulit. Manusia sering terserang oleh bakteri yang bisa berubah menjadi patogen seperti Clostridia, Enterobacteria, dan Bacteroides, melalui saluran pencernaan. Hal ini terutama disebabkan oleh makanan yang tidak bersih. Berbagai macam food and water born disease dapat menyebabkan berbagai macam penyakit yang menyerang traktus gastrointestinal, karena tidak seimbangnya mikrobiota yang terdapat dalam traktus gastrointestinal.

Lactobacillus dan Bifidobacteria dapat memproduksi bahan penetralisir toksin

yang dihasilkan bakteri patogen. Pertahanan yang kuat pada traktus gastrointestinal sangatlah diperlukan. Salah satunya dengan mempertahankan bakteri asam laktat dan menguntungkan, yang dapat berkompetisi kuat dengan bakteri patogen yang menyerang manusia.

Lactobacillus dapat kita peroleh dari sumber hewani dan nabati. Sumber

hewani misalnya produk-produk susu; sumber nabati misalnya yang biasanya dipakai untuk asinan buah dan sayuran.

2

1.2Identifikasi Masalah

Apakah Lactobacillus rhamnosus GG, Bifidobacterium Bb12,

Lactobacillus plantarum IS-10506, dan Lactobacillus plantarum IS-20506 dapat

menggantikan posisi dan menghambat adhesi Bacteroides vulgatus pada mukus saluran pencernaan?

1.3Maksud dan Tujuan

Maksud penelitian ini adalah, mengetahui kemampuan Lactobacillus

rhamnosus GG, Bifidobacterium Bb12, Lactobacillus plantarum IS-10506, dan Lactobacillus plantarum IS-20506 dapat menggantikan posisi dan menghambat Bacteroides vulgatus.

Tujuan penelitian ini adalah mengetahui persentase penggantian posisi dan penghambatan adhesi bakteri patogen Bacteroides vulgatus pada mukus, oleh bakteri-bakteri probiotik; Lactobacillus rhamnosus GG, Bifidobacterium Bb12,

Lactobacillus plantarum IS-10506, dan Lactobacillus plantarum IS-20506.

1.4Kerangka Pemikiran

Usus manusia dihuni oleh bakteri yang jumlahnya lebih banyak daripada jumlah sel yang ada dalam tubuh manusia. Sekitar 400 spesies bakteri yang berlainan tinggal di dalam saluran pencernaan manusia. Jenis bakteri yang terdapat dalam usus terdiri dari bakteri “bersahabat” dan “tidak bersahabat”. (Sorokie,2004; Salminen, dkk., 2004)

Bakteri probiotik, yang juga termasuk dalam kelompok bakteri asam laktat, dapat menjaga keseimbangan ekosistem dalam traktus gastrointestinal.

Penggunaan bakteri probiotik untuk mencegah infeksi dengan cara menggantikan posisi dan menghambat adhesi bakteri patogen.

3

1.5 Hipotesis Penelitian

Bakteri probiotik Lactobacillus rhamnosus GG, Bifidobacterium Bb12, Lactobacillus plantarum IS-10506 dan Lactobacillus plantarum IS-20506 dapat

menggantikan posisi dan menghambat adhesi bakteri patogen Bacteroides

vulgatus pada mukus pencernaan .

1.6 Metode Penelitian

Desain penelitian menggunakan metode mucus adhesion assay dengan sifat penelitian eksperimental laboratorik prospektif. Pengujian dilakukan dengan menghitung persentase penghambatan adhesi patogen oleh bakteri probiotik pada mukus pencernaan yang didapat dari orang sehat, bakteri patogen yang telah diradiolabel dibaca dengan menggunakan liquid scintilation counter untuk penghitungan sisa jumlah bakteri patogen.

Metode statistik yang digunakan yaitu ANAVA satu arah pada taraf kepercayaan 95%.

1.7 Lokasi dan Waktu

Penelitian dilakukan di laboratorium Functional Foods Forum, University

of Turku, Itäinen Pitkäkatu,Turku, Finland. Mulai dari bulan April hingga Juni

2007.

BAB V

KESIMPULAN DAN SARAN

5.1 Kesimpulan

Berdasarkan hasil penelitian dapat ditarik kesimpulan sebagai berikut:

1. Bakteri-bakteri probiotik Lactobacillus rhamnosus GG, dan Lactobacillus

plantarum IS-20506 dapat menggantikan posisi adhesi bakteri patogen Bacteroides vulgatus. Sedangkan Bifidobacterium Bb12, Lactobacillus plantarum IS-10506 tidak dapat menggantikan posisi adhesi Bacteroides vulgatus pada mukus.

2. Bakteri-Bakteri probiotik Lactobacillus rhamnosus GG, Bifidobacterium Bb12, Lactobacillus plantarum IS-10506, dan Lactobacillus plantarum IS-20506 tidak dapat menghambat adhesi bakteri patogen Bacteroides

vulgatus pada mukus.

5.2 Saran

Beberapa saran penelitian dapat diajukan sebagai, berikut:

• Penelitian lebih lanjut untuk mengetahui mekanisme molekuler pergantian tempat dan penghambatan bakteri probiotik atas bakteri patogen.

• Penelitian keuntungan dan kerugian dari peningkatan adhesi bakteri patogen pada mukus.

• Penelitian menggunakan kombinasi dari beberapa bakteri probiotik dan patogen.

• Penelitian lebih lanjut perkembangan dan pertumbuhan bakteri probiotik dari adhesi pada mukus pencernaan.

• Penelitian menggunakan bakteri patogen penyebab infeksi umum yang terjadi di Indonesia.

• Penelitian pada manusia langsung (in vivo).

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Vesterlund S., Paltta J., Karp M., Ouwehand A.C. 2005. Adhesion of bacteria to resected human colonic tissue: Quantitative analysis of bacterial

adhesion and viability. Research in Microbiology 156 (2005) 238-244.

Vesterlund S., Karp M., Salminen S., and Ouwehand A.C. 2006. Staphylococcus

aureus adheres to human intestinal mucus but can be displaced by

certain lactic acid bacteria. Microbiology 152 (2006), 1819-1826.

What Are Bacteroides?. 2007. http://borg.med.ecu.edu/~webpage/about.html .,

14th May, 2007.

Wikipedia. 2007. Bacteroides, http://en.wikipedia.org/wiki/bacteroides., 14th may, 2007.

Wikipedia. 2007. Lactobacillus_rhamnosus,

http://en.wikipedia.org/wiki/Lactobacillus_rhamnosus., 28th August, 2007.

Zhou J.S., Pillidge C.J., Gopal P.K., Gill H.S. 2005. Antibiotic susceptibility

profiles of new probiotic Lactobacillus and Bifidobacterium strains.

International Journal of Food Microbiology 98 (2005) 211-217.

BAB V

KESIMPULAN DAN SARAN

5.1 Kesimpulan

Berdasarkan hasil penelitian dapat ditarik kesimpulan sebagai berikut:

1. Bakteri-bakteri probiotik Lactobacillus rhamnosus GG, dan Lactobacillus plantarum IS-20506 dapat menggantikan posisi adhesi bakteri patogen Bacteroides vulgatus. Sedangkan Bifidobacterium Bb12, Lactobacillus plantarum IS-10506 tidak dapat menggantikan posisi adhesi Bacteroides vulgatus pada mukus.

2. Bakteri-Bakteri probiotik Lactobacillus rhamnosus GG, Bifidobacterium Bb12, Lactobacillus plantarum IS-10506, dan Lactobacillus plantarum IS-20506 tidak dapat menghambat adhesi bakteri patogen Bacteroides vulgatus pada mukus.

5.2 Saran

Beberapa saran penelitian dapat diajukan sebagai, berikut:

• Penelitian lebih lanjut untuk mengetahui mekanisme molekuler pergantian tempat dan penghambatan bakteri probiotik atas bakteri patogen.

• Penelitian keuntungan dan kerugian dari peningkatan adhesi bakteri patogen pada mukus.

• Penelitian menggunakan kombinasi dari beberapa bakteri probiotik dan patogen.

• Penelitian lebih lanjut perkembangan dan pertumbuhan bakteri probiotik dari adhesi pada mukus pencernaan.

• Penelitian menggunakan bakteri patogen penyebab infeksi umum yang terjadi di Indonesia.

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