Pengaruh Lama Fermentasi Terhadap Jumlah Koloni Bakteri Asam Laktat Dalam Soyghurt Dan Efektifitas Pada Penyembuhan Gastritis Lambung Mencit (Mus musculus L) Yang Diinduksi Dengan Aspirin

  L-2

LAMPIRAN B

  Lampiran B. Komposisi Media (Karna, et al., 2007) Media MRSA ( De man Rogosa Sharpe Agar) Pepton 10 gram Beef extrac 10 gram Yeast extract 5 gram K

  2 HPO 4 2 gram Ammonium sitrat 2 gram Glukosa 2 gram Natrium acetat 20 gram MgSO

  4 .7H

  2 O 0,58 gram MnSO .4H O 0,28 gram

  4

  2 Agar 15 gram Akuades 1 liter

Lampiran C. Cakupan Kegiatan Penelitian

  Pembuatan starter youghurt Inokulasi Inkubasi suhu 40

• – 45 C Soyghurt I 4 jam

  Soyghurt III 8 jam 8 jam Soyghurt II

  6 jam 6 jam Pembuatan susu kedelai kultur

  Histopatologi Hewan Percobaan Hitung Koloni Lampiran D . Diagram Alir Pembuatan Starter Syoghurt (Koswara, 2006) 16%

  Susu bubuk dilarutkan dengan air masak hingga mencapai total solid Dipanaskan 80 C, 30 menit Ditambahkan gula pasir 2%

  Diinokulasi dengan kultur starter komersial 5% murni

dikocok

  

Diinkubasi pada suhu 40-45 C selama 4-8 jam

Kultur starter Lampiran E. Diagram Alir Pembuatan Susu Kedelai (Yudhi, 2008).

  1 Kg kacang kedelai

Direndam 8 jam dalam air yang mengandung 0.5% NAHCO

  3 Cuci, ditiriskan

Direbus dalam air mendidih selama 30 menit

Ditiriskan, dikupas kulit kacang kedelai

  

Biji kacang kedelai diblender dengan air panas (90°C) dengan perbandingan

1:6 dari berat kering kedelai

Disaring Filtrat

  

Dipanaskan 80-90°C, 30 menit Didinginkan sampai suhu 40°C Susu kedelai Lampiran F. Alir Pembuatan Soyghurt (Koswara, 2006) Susu kedelai hangat 40 C

  Ditambah susu bubuk Ditambah Inokulasi

  Diaduk sampai mencapai total gula 2% dengan sampai kultur solid 16% larut stater 2%

  Soyghurt I (4 jam) Dimasukkan ke dalam

  

Diinkubasi pada

wadah Soyghurt

suhu 37- 40  C

tertutup yang

  II (6 jam) selama berpori sesuai dengan

  Soyghurt perlakuan

  III (8 jam)

Lampiran G. Tabel Jumlah Koloni L. bulgaricus dan S. thermophillus pada -7 Pengenceran 10

  Jumlah Koloni (CFU/ml) Ulangan Jenis Isolat Rata-rata

  1

  2

  3

  43 Lactobacillus bulgaricus

  43

  39

  47 114 Streptococcus thermophillus 112 109 122

Lampiran H. Tabel Identifikasi Fermentasi Karbohidrat Beberapa Genus

Lactobacillus Ber dasarkan Bergey’s

  lus ckii annii hi icus cus ius ti s hm ti tar lbrue idop

ic nsenii

lve li ac ac

  1. L. de

  2. L. le

  3. L. je

  4. L. l

  5. L. bulgar

  6. L. he 7. L.

  8. L.sa

  1. Amigdalin + + - - - - + -

  2. Arabinose

• - - - - - -

  3. Collobiose - - + + - - - +

• 4. Fruetose

  # + + + + + +

  5. Galactose # -w + + + + +

• + 6. Glucose (acid) + + + + + + + +

  7. Glucose (gas) _ _ - - - - - -

  8. Gluconate - _ - _ - - - -

  9. Lactose

• - + + + +

  10. Maltose + - + + d + +

• + 11. Manitol
• _ - -

  12 Mannose + + - + + + + #

  13 Melezitose - - - - - - - -

• 14 Melibiose - - - - - d +

  15. Raffinose - - - + - d

• - -

  16. Rhamnose - - - - - * - -

  17. Ribose - - - - - - - -

  18. Salicin + - * + - + - +

  19 Sorbitol - - - + - - - -

  20. Sucrose

• + +
• 21. Trehnalose
• + +

  22. Xylose - - - - - - - -

  23. Esculin # * - + - - + + The symbol used are : + = positive reaction by 90% or more strains; d = some strains +, others

• – (about 89-11% positive);
• = negative reaction by most strains (90% or more); ( ) = delayed reaction; w = weak reation; ≠ = weak, slow or negative; * = see text; 0 = not tested. Combination of symbols, e.g.(-) = rarely positive, and then slowly; -w = negative or weak reaction.

Fermentasi Karbohidrat Beberapa Spesies S. Termophillus Berdasarkan Bergey’s

  Glucose Lactosa Fructosa Maltosa Manitol Sucrose

• S. Thermophillus

Lampiran I . Hasil Uji Statistik Peningkatan Berat Badan Mencit pada Hari ke-0 dan Hari Ke-24 pada Masing-masing Kelompok Perlakuan Tests of Normal ity

  a

  Kolmogorov -Smirnov Shapiro-Wilk kelompok St at ist ic df Sig. St at ist ic df Sig. Berat Badan K

  .211 5 .200* .918 5 .514 P1

  .225 5 .200* .909 5 .464 P2

  .225 5 .200* .912 5 .479 P3 .283 5 .200* .855 5 .212 *.

  This is a lower bound of the true signif icance.

  a.

  Lillief ors Signif icance Correction

  Test of Homogeneity of Variance

  Lev ene Stat istic df 1 df 2 Sig. Berat Badan Based on Mean

  3.112

  3 16 .056 Based on Median 1.558

  3 16 .238 Based on Median and

  1.558 3 10.299 .258 with adjusted df Based on trimmed mean

  3.069

  3 16 .058

Oneway ANOVA

  Berat Badan Sum of Squares df Mean Square F Sig.

  Between Groups 69.916 3 23.305 27.301 .000

  Within Groups 13.658 16 .854 Total

  83.574

  19

Post Hoc Tests Multi ple Comparisons

  Dependent Variable: Berat Badan Bonf erroni

  4.74000* .58434 .000 2.9821 6.4979 4.33200* .58434 .000 2.5741 6.0899 2.53000* .58434 .003 .7721 4.2879

  (J) kelompok P1 P2 P3 K P2 P3 K P1 P3 K P1 P2

  (I) kelompok K P1 P2 P3

  Mean Dif f erence

  (I-J) St d. Error Sig. Lower Bound Upper Bound 95% Conf idence Interv al The mean dif f erence is signif icant at the . 05 lev el.

• 4.74000* .58434 .000 -6.4979 -2.9821
• .40800 .58434 1.000 -2.1659 1.
• 2.21000* .58434 .010 -3.9679 -.4521
• 4.33200* .58434 .000 -6.0899 -2.5741

  .40800 .58434 1.000 -1.3499 2.1659

• 1.80200* .58434 .043 -3.5599 -.0441
• 2.53000* .58434 .003 -4.2879 -.7721

  2.21000* .58434 .010 .4521 3.9679 1.80200* .58434 .043 .0441 3.5599

• .

Lampiran J . Data-Data Bobot Lambung, Makroskopis Lambung dan Histopatologi Lambung Mencit pada Hari ke-24 pada Masing-masing Kelompok Perlakuan

  Histopatologi Makroskopis Bobot Lambung lambung lambung

Kelompok Notasi Notasi Notasi

  Rata-rata±SD Rata-rata±SD Rata-rata±SD K 0,392±0,102 a 0,60±0,55 a 0,00±0,00 a P1 0,346±0,081 a 0,80±0,45 ab 0,80±0,80 ab P2 0,328±0,044 a 2,00±1,00 bc 1,20±1,20 b P3 b c

  0,486±0,076 2,80±0,45 2,80±2,80 c

Lampiran L. Hasil Uji Statistik Peningkatan Bobot Lmabung dan Histopatologi Lambung Mencit pada Hari Ke-24 pada Masing-masing Kelompok Perlakuan Kruskal-Wallis Test Ranks

  kelompok N Mean Rank Berat Lambung K

  5

  10.20 P1

  5

  7.90 P2

  5

  7.00 P3

  5

  16.90 Total

  20 Hispatologi Lambung K

  5

  4.80 P1

  5

  8.20 P2

  5

  12.40 P3

  5

  16.60 Total

  20 Makroskopis Lambung K

  5

  5.00 P1

  5

  9.00 P2

  5

  10.40 P3

  5

  17.60 Total

  20 a,b Test Statistics

  Berat Hispatologi Makroskopis Lambung Lambung Lambung

  Chi-Square 8.619 12.764 13.363 df

  3

  3

  3 Asy mp. Sig. .035 .005 .004 a. Kruskal Wallis Test b. Grouping Variable: kelompok

Mann-Whitney Test

  5

  Berat Lambung

  Hispatologi Lambung

  Makroskopis Lambung Not corrected f or ties.

  10.000 6.000 5.000 25.000 21.000 20.000

  Test Statistics b

  Hispatologi Lambung Makroskopis Lambung N Mean Rank Sum of Ranks

  10 kelompok K P1 Total K P1 Total K P1 Total Berat Lambung

  35.00

  7.00

  5

  20.00

  4.00

  10

  

Ranks

  34.00

  6.80

  5

  21.00

  4.20

  5

  10

  25.00

  5.00

  5

  30.00

  6.00

  5

  Mann-Whitney U Wilcoxon W Z Asy mp. Sig. (2-tailed) Exact Sig. [2*(1-tailed Sig.)]

• .529 -1.678 -1.936

  .151

  18.00

  N Mean Rank Sum of Ranks

  Berat Lambung Hispatologi Lambung Makroskopis Lambung

  10 kelompok K P2 Total K P2 Total K P2 Total

  35.00

  7.00

  5

  20.00

  4.00

  5

  10

  37.00

  7.40

  5

  3.60

  a

  5

  10

  22.00

  4.40

  5

  33.00

  6.60

  5

  Ranks

  Grouping Variable: kelompok b.

  a.

  .597 .093 .053 .690

  a

  .222

  a

  b Test Statistics

  Berat Hispatologi Makroskopis Lambung Lambung Lambung

  Mann-Whitney U 7.000 3.000 5.000

  Wilcoxon W 22.000 18.000 20.000

  Z

• 1.163 -2.132 -1.964 Asy mp. Sig. (2-tailed) .245 .033 .050

  a a a

  Exact Sig. [2*(1-tailed .310 .056 .151

  Sig.)] a.

  Not corrected f or ties.

  b.

  Grouping Variable: kelompok

  Ranks

  kelompok N Mean Rank Sum of Ranks Berat Lambung K

  5

  3.60

  18.00 P3

  5

  7.40

  37.00 Total

  10 Hispatologi Lambung K

  5

  3.00

  15.00 P3

  5

  8.00

  40.00 Total

  10 Makroskopis Lambung K

  5

  3.00

  15.00 P3

  5

  8.00

  40.00 Total

  10

  b Test Statistics

  Berat Hispatologi Makroskopis Lambung Lambung Lambung

  Mann-Whitney U 3.000 .000 .000

  Wilcoxon W 18.000 15.000 15.000 Z

• 2.009 -2.739 -2.887 Asy mp. Sig. (2-tailed)

  .045 .006 .004

  a a a

  Exact Sig. [2*(1-tailed .056 .008 .008

  Sig.)] a.

  Not corrected f or ties.

  b.

  Grouping Variable: kelompok

  Ranks

  kelompok N Mean Rank Sum of Ranks Berat Lambung P1

  5

  5.70

  28.50 P2

  5

  5.30

  26.50 Total

  10 Hispatologi Lambung P1

  5

  4.30

  21.50 P2

  5

  6.70

  33.50 Total

  10 Makroskopis Lambung P1

  5

  4.90

  24.50 P2

  5

  6.10

  30.50 Total

  10

  b Test Statistics

  Berat Hispatologi Makroskopis Lambung Lambung Lambung

  Mann-Whitney U 11.500 6.500 9.500

  Wilcoxon W 26.500 21.500 24.500 Z

• .210 -1.423 -.671 Asy mp. Sig. (2-tailed)

  .834 .155 .502

  a a a

  Exact Sig. [2*(1-tailed .841 .222 .548

  Sig.)] a.

  Not corrected f or ties.

  b.

  Grouping Variable: kelompok

  Ranks

  kelompok N Mean Rank Sum of Ranks Berat Lambung P1

  5

  3.20

  16.00 P3

  5

  7.80

  39.00 Total

  10 Hispatologi Lambung P1

  5

  3.10

  15.50 P3

  5

  7.90

  39.50 Total

  10 Makroskopis Lambung P1

  5

  3.10

  15.50 P3

  5

  7.90

  39.50 Total

  10 Grouping Variable: kelompok b.

  a.

  Grouping Variable: kelompok b.

  a

  .008

  a

  .008

  a

  Mann-Whitney U Wilcoxon W Z Asy mp. Sig. (2-tailed) Exact Sig. [2*(1-tailed Sig.)]

  Berat Lambung

  Hispatologi Lambung

  Makroskopis Lambung Not corrected f or ties.

  a.

  Test Statistics b

  1.000 .500 .500 16.000 15.500 15.500

  1.500 6.500 1.500 16.500 21.500 16.500

  .021 .155 .014 .016

  a

  .222

  a

  .016

  a

  Mann-Whitney U Wilcoxon W Z Asy mp. Sig. (2-tailed) Exact Sig. [2*(1-tailed Sig.)]

  Berat Lambung

  Hispatologi Lambung

  Makroskopis Lambung Not corrected f or ties.

  .016 .007 .009 .016

  Test Statistics b

• 2.402 -2.683 -2.612

  21.50

  Ranks

  5

  3.30

  16.50

  5

  7.70

  38.50

  10

  5

  4.30

  5

  N Mean Rank Sum of Ranks

  6.70

  33.50

  10

  5

  3.30

  16.50

  5

  7.70

  38.50

  10 kelompok P2 P3 Total P2 P3 Total P2 P3 Total

  Berat Lambung Hispatologi Lambung Makroskopis Lambung

• 2.305 -1.423 -2.460

Lampiran K. Persentase Ketebalan Mukosa Lambung Mencit pada Hari ke 24 pada Masing-masing Kelompok Perlakuan. Tests of Normal ity

  a

  Kolmogorov -Smirnov Shapiro-Wilk kelompok Statistic df Sig. Statistic df Sig. % Ketebalan Mukosa K

  .298 5 .169 .851 5 .197 P1

  .170 5 .200* .965 5 .841 P2

  .192 5 .200* .939 5 .661 P3 .234 5 .200* .858 5 .221 *.

  This is a lower bound of the true significance.

  a.

  Lillief ors Significance Correction

  

Test of Homogeneity of Variance

  Lev ene Stat istic df 1 df 2 Sig. % Ketebalan Mukosa Based on Mean

  2.617

  3 16 .087 Based on Median

  1.795

  3 16 .189 Based on Median and

  1.795 3 8.011 .226 with adjusted df Based on trimmed mean

  2.442

  3 16 .102

Oneway ANOVA

  % Ketebalan Mukosa Sum of Squares df Mean Square F Sig. Between Groups 2325.896

  3 775.299 5.276 .010 Within Groups 2351.249 16 146.953 Total 4677.145

  19

Post Hoc Tests Multi ple Comparisons

  Dependent Variable: % Ket ebalan Mukosa Bonf erroni

• 2.57000 7.66689 1.000 -25.6346 20.4946 7.94000 7.66689 1.000 -15.1246 31.0046 25.03200* 7.66689 .029 1.9674 48.0966 2.57000 7.66689 1.000 -20.4946 25.6346 10.51000 7.66689 1.000 -12.5546 33.5746 27.60200* 7.66689 .014 4.5374 50.6666
• 7.94000 7.66689 1.000 -31.0046 15.1246

  P2 P3 K P1 P3 K P1 P2 (I) kelompok K P1 P2 P3

  Mean Dif f erence (I-J) St d. Error Sig. Lower Bound Upper Bound 95% Conf idence Interv al

• 10.51000 7.66689 1.000 -33.5746 12.5546 17.09200 7.66689 .243 -5.9726 40.1566
• 25.03200* 7.66689 .029 -48.0966 -1.9674
• 27.60200* 7.66689 .014 -50.6666 -4.5374
• 17.09200 7.66689 .243 -40.1566 5.9726 (J) kelompok P1 P2 P3 K
• .

  The mean dif f erence is signif icant at the . 05 lev el.

  Tabel. Rata-rata Ketebalan Mukosa (%) Kelompok Rata-rata Notasi K

  57,67±7,28 a P1 60,24±9,58 a P2 49,73±20,40 ab P3

  32,64±5,18 b

Lampiran L. Dokumentasi Penelitian

  B A Gambar 5. A (Kacang kedelai) dan B (Stater komersial).

  Gambar 6. Soyghurt Inkubasi 4,6 dan 8 jam (kiri) dan Kultur Starter (kanan) Gambar 7. Bakteri Asam Laktat pada Soyghurt Inkubasi 4, 6 dan 8 jam

• 6

  Pengenceran 10 (dari kiri ke kanan) A B Gambar 8. Uji Pewarnaan Gram, A (L. bulgaricus) dan B (S. termophillus) Gambar 9. Uji Fermentasi Karbohidrat A (L. Bulgaricus) dan B ( S. termophillus) Gambar 10. Alat untuk Pemeriksaan Kultur Bakteri Asam Laktat A (Inkubator) dan B (Auto clave) Gambar 11. Alat untuk Pemeriksaan Histopatologi A (Automatic Tissue Processor/Leica TP 1020), B (Tissue Embedding /Leica EG 1160),

  C (Microtome/Leica RM 2245).

  A B C A B A

  B Gambar 12. Mencit Yang Digunakan untuk Penelitian Gambar 13. Blok Paraffin , slide dan Pewarnaan Haematoxyline - Eosin