5.1 Simpulan
1. Pemberian probiotik L. fermentum selama 2-3 minggu dapat meningkatkan kandungan enzim Cu,Zn-SOD di jaringan hati tikus. Sedangkan pemberian probiotik L. plantarum hanya mampu mempertahankan kandungan enzim antioksidan tersebut seperti pada kondisi kontrol negatif.
2. Pada tikus yang dipapar EPEC, pemberian probiotik L. fermentum
selama 2-3 minggu dapat meningkatkan kandungan enzim Cu,Zn-SOD di jaringan hati tikus. Sedangkan pemberian L. plantarum hanya mampu mempertahankan kandungan enzim antioksidan Cu,Zn-SOD pada minggu kedua, lalu kandungan enzim antioksidan tersebut menurun pada minggu ketiga.
3. L. fermentum lebih baik dari pada L. plantarum dalam hal kemampuan meningkatkan kandungan enzim Cu,Zn-SOD di jaringan hati tikus, baik pada kelompok yang dipapar EPEC maupun tidak.
5.2 Saran
1. Perlu dilakukan penelitian mengenai pemanfaatan L. fermentum dan
L. plantarum ini sebagai salah satu komponen dalam produk pangan fungsional dan pakan hewan.
2. Perlu dilakukan penelitian mengenai efek L. fermentum dan
L. plantarum dalam mencegah infeksi patogen lain seperti Salmonella
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Lampiran 1 Tahapan Sampling dan Pemrosesan Jaringan Hati Tikus
pengambilan jaringan hati
fiksasi
dehidrasi
penjernihan (clearing)
blok potong dengan mikrotom
alkohol 70%, 80%, 90%, dan 95% (@ 24 jam)
dehidrasi alkohol absolut I, II, dan
III (@ 1 jam)
xylol I, II, dan III (@ 1 jam)
infiltrasi parafin parafin I, II, dan III (@ 1 jam)
embedding
hasil potongan dibentangkan dalam akuades suhu ruang
bentangkan dalam akuades (37 °C)
tikus dibunuh, darah dikeluarkan
celupkan ke NaCl fisiologis
sayatan jaringan hati di tempelkan pada objek gelas
Lampiran 2 Tahapan Pewarnaan Imunohistokimia untuk Analisis Kandungan Cu, Zn-SOD di Jaringan Hati Tikus
xylol II, II, dan I (@ 3 menit) deparafinisasi rehidrasi alkohol 95%, 90%, 80% , dan 70% (@ 3 menit) rehidrasi
alkohol absolut III, II, dan I (@ 3 menit)
dicuci dengan air kran (3-5menit) dan MilliQ (5-10 menit)
dicuci dengan PBS 3x (@ 5-10menit) dicelupkan ke dalam 0.5 mL H2O2 dalam 50 mL
metanol (15 menit, suasana gelap)
diinkubasi dengan 50-60 µl antibodi primer/monoklonal Cu, Zn-SOD (48 jam, 4 °C) dicuci dengan MilliQ 2x (@10-15 menit) dan PBS
2x (@ 5-10 menit)
diinkubasi dengan 50-60 µl serum normal (60 menit, 37 °C)
diinkubasi dengan 50-60 µl antibodi sekunder Dako Envision Peroxidase System (60 menit, 37 °C,
suasana gelap)
dehidrasi alkohol 70%, 80%, 90%, dan
95% (@ 1-3 menit)
dijernihkan ulang (clearing)
entelan
xylol I, II, dan III (@ 5 menit)
di-mounting
dehidrasi alkohol absolut I, II, dan III (@ 1-3 menit) dicuci dengan PBS 3x (@ 5-10 menit)
diteteskan larutan DAB (30 menit)
dicek di bawah mikroskop
diamati di bawah mikroskop counterstain dengan pewarna
Lampiran 3 Uji statistik (Anova) dengan SPSS untuk jumlah inti sel hati tikus pada terminasi hari ke-8 dengan intensitas kandungan Cu,Zn-SOD positif kuat (+++)
Univariate Analysis of Variance
Tests of Between-Subjects Effects
Dependent Variable: Jumlah inti sel hati
Source
Type III Sum
of Squares df Mean Square F Sig.
Model 68403.417a 11 6218.492 1.775E3 .000
Perlakuan 10.250 5 2.050 .585 .711
Ulangan 10.917 5 2.183 .623 .683
Error 87.583 25 3.503
Total 68491.000 36
Lampiran 4 Uji statistik (Anova) dengan SPSS untuk jumlah inti sel hati tikus pada terminasi hari ke-8 dengan intensitas kandungan Cu,Zn-SOD positif sedang (++)
Univariate Analysis of Variance
Tests of Between-Subjects Effects
Dependent Variable: Jumlah inti sel hati Source Type III Sum of
Squares
df Mean Square F Sig.
Model 134844.306a 11 12258.573 3.618E3 .000
Perlakuan 15.806 5 3.161 .933 .477
Ulangan 17.139 5 3.428 1.012 .432
Error 84.694 25 3.388
Total 134929.000 36
Lampiran 5 Uji statistik (Anova) dengan SPSS untuk jumlah inti sel hati tikus pada terminasi hari ke-8 dengan intensitas kandungan Cu,Zn-SOD positif lemah (+)
Univariate Analysis of Variance
Tests of Between-Subjects Effects
Dependent Variable: Jumlah inti sel hati
Source
Type III Sum of
Squares df Mean Square F Sig.
Model 153.639a 11 13.967 13.768 .000
Perlakuan 21.806 5 4.361 4.299 .006
Ulangan 7.139 5 1.428 1.407 .256
Error 25.361 25 1.014
Total 179.000 36
a. R Squared = ,858 (Adjusted R Squared = ,796)
Post Hoc Tests Perlakuan
Homogeneous Subsets
Jumlah inti sel hati
Duncan Perlakuan N Subset 1 2 T1F 6 .8333 T1E 6 1.0000 T1D 6 1.5000 1.5000 T1B 6 2.5000 T1C 6 2.6667 T1A 6 2.6667 Sig. .290 .077
Means for groups in homogeneous subsets are displayed. Based on observed means.
Lampiran 6 Uji statistik (Anova) dengan SPSS untuk jumlah inti sel hati tikus pada terminasi hari ke-8 dengan intensitas kandungan Cu,Zn-SOD negatif (-)
Univariate Analysis of Variance
Tests of Between-Subjects Effects
Dependent Variable: Jumlah inti sel hati
Source
Type III Sum of
Squares df Mean Square F Sig.
Model 69.222a 11 6.293 10.646 .000
Perlakuan 9.889 5 1.978 3.346 .019
Ulangan .556 5 .111 .188 .964
Error 14.778 25 .591
Total 84.000 36
a. R Squared = ,824 (Adjusted R Squared = ,747)
Post Hoc Tests Perlakuan
Homogeneous Subsets
Jumlah inti sel hati
Duncan Perlakuan N Subset 1 2 T1D 6 .6667 T1F 6 .6667 T1E 6 1.1667 T1A 6 1.5000 1.5000 T1B 6 1.5000 1.5000 T1C 6 2.1667 Sig. .104 .168
Means for groups in homogeneous subsets are displayed. Based on observed means.
Lampiran 7 Uji statistik (Anova) dengan SPSS untuk jumlah inti sel hati tikus pada terminasi hari ke-15 dengan intensitas kandungan Cu,Zn-SOD positif kuat (+++)
Univariate Analysis of Variance
Tests of Between-Subjects Effects
Dependent Variable: Jumlah inti sel hati
Source
Type III Sum of
Squares df Mean Square F Sig.
Model 118515.222a 11 10774.111 2.029E3 .000
Perlakuan 28104.222 5 5620.844 1.058E3 .000
Ulangan 10.556 5 2.111 .397 .846
Error 132.778 25 5.311
Total 118648.000 36
a. R Squared = ,999 (Adjusted R Squared = ,998)
Post Hoc Tests Perlakuan
Homogeneous Subsets
Jumlah inti sel hati
Duncan Perlakuan N Subset 1 2 3 4 T2F 6 12.8333 T2D 6 37.1667 T2B 6 38.0000 T2A 6 39.8333 T2C 6 75.5000 T2E 6 97.3333 Sig. 1.000 .068 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means.
Lampiran 8 Uji statistik (Anova) dengan SPSS untuk jumlah inti sel hati tikus pada terminasi hari ke-15 dengan intensitas kandungan Cu,Zn-SOD positif sedang (++)
Univariate Analysis of Variance
Tests of Between-Subjects Effects
Dependent Variable: Jumlah inti sel hati
Source
Type III Sum of
Squares df Mean Square F Sig.
Model 148860.750a 11 13532.795 2.638E3 .000
Perlakuan 24595.583 5 4919.117 958.892 .000
Ulangan 8.917 5 1.783 .348 .879
Error 128.250 25 5.130
Total 148989.000 36
a. R Squared = ,999 (Adjusted R Squared = ,999)
Post Hoc Tests Perlakuan
Homogeneous Subsets
Jumlah inti sel hati
Duncan Perlakuan N Subset 1 2 3 4 5 T2E 6 15.0000 T2C 6 35.6667 T2A 6 66.3333 T2B 6 66.8333 T2D 6 73.8333 T2F 6 94.8333 Sig. 1.000 1.000 .705 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means.
Lampiran 9 Uji statistik (Anova) dengan SPSS untuk jumlah inti sel hati tikus pada terminasi hari ke-15 dengan intensitas kandungan Cu,Zn-SOD positif lemah (+)
Univariate Analysis of Variance
Tests of Between-Subjects Effects
Dependent Variable:Jumlah inti sel hati
Source
Type III Sum of
Squares df Mean Square F Sig.
Model 164.639a 11 14.967 9.754 .000
Perlakuan 45.806 5 9.161 5.970 .001
Ulangan 15.472 5 3.094 2.017 .111
Error 38.361 25 1.534
Total 203.000 36
a. R Squared = ,811 (Adjusted R Squared = ,728)
Post Hoc Tests Perlakuan
Homogeneous Subsets
Jumlah inti sel hati
Duncan Perlakuan N Subset 1 2 3 T2A 6 .3333 T2E 6 .6667 T2D 6 1.3333 1.3333 T2C 6 1.5000 1.5000 T2B 6 2.8333 2.8333 T2F 6 3.5000 Sig. .147 .057 .360
Means for groups in homogeneous subsets are displayed. Based on observed means.
Lampiran 10 Uji statistik (Anova) dengan SPSS untuk jumlah inti sel hati tikus pada terminasi hari ke-15 dengan intensitas kandungan Cu,Zn-SOD negatif (-)
Univariate Analysis of Variance
Tests of Between-Subjects Effects
Dependent Variable: Jumlah inti sel hati
Source
Type III Sum of
Squares df Mean Square F Sig.
Model 78.639a 11 7.149 8.778 .000
Perlakuan 25.806 5 5.161 6.337 .001
Ulangan 1.472 5 .294 .362 .870
Error 20.361 25 .814
Total 99.000 36
a. R Squared = ,794 (Adjusted R Squared = ,704)
Post Hoc Tests Perlakuan
Homogeneous Subsets
Jumlah inti sel hati
Duncan Perlakuan N Subset 1 2 T2A 6 .5000 T2B 6 .6667 T2E 6 .8333 T2C 6 .8333 T2D 6 1.3333 T2F 6 3.0000 Sig. .164 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means.
Lampiran 11 Uji statistik (Anova) dengan SPSS untuk jumlah inti sel hati tikus pada terminasi hari ke-22 dengan intensitas kandungan Cu,Zn-SOD positif kuat (+++)
Univariate Analysis of Variance
Tests of Between-Subjects Effects
Dependent Variable: Jumlah inti sel hati
Source
Type III Sum of
Squares df Mean Square F Sig.
Model 93797.556a 11 8527.051 886.593 .000
Perlakuan 25793.889 5 5158.778 536.379 .000
Ulangan 56.556 5 11.311 1.176 .349
Error 240.444 25 9.618
Total 94038.000 36
a. R Squared = ,997 (Adjusted R Squared = ,996)
Post Hoc Tests Perlakuan
Homogeneous Subsets
Jumlah inti sel hati
Duncan Perlakuan N Subset 1 2 3 4 5 T3F 6 7.1667 T3D 6 26.0000 T3B 6 38.0000 T3A 6 41.3333 T3C 6 54.5000 T3E 6 93.6667 Sig. 1.000 1.000 .074 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means.
Lampiran 12 Uji statistik (Anova) dengan SPSS untuk jumlah inti sel hati tikus pada terminasi hari ke-22 dengan intensitas kandungan Cu,Zn-SOD positif sedang (++)
Univariate Analysis of Variance
Tests of Between-Subjects Effects
Dependent Variable: Jumlah inti sel hati
Source
Type III Sum of
Squares df Mean Square F Sig.
Model 165322.222a 11 15029.293 2.323E3 .000
Perlakuan 19627.889 5 3925.578 606.631 .000
Ulangan 24.889 5 4.978 .769 .581
Error 161.778 25 6.471
Total 165484.000 36
a. R Squared = ,999 (Adjusted R Squared = ,999)
Post Hoc Tests Perlakuan
Homogeneous Subsets
Jumlah inti sel hati
Duncan Perlakuan N Subset 1 2 3 4 5 6 T3E 6 18.6667 T3C 6 51.1667 T3A 6 66.5000 T3B 6 74.8333 T3D 6 81.6667 T3F 6 88.8333 Sig. 1.000 1.000 1.000 1.000 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means.
Lampiran 13 Uji statistik (Anova) dengan SPSS untuk jumlah inti sel hati tikus pada terminasi hari ke-22 dengan intensitas kandungan Cu,Zn-SOD positif lemah (+)
Univariate Analysis of Variance
Tests of Between-Subjects Effects
Dependent Variable: Jumlah inti sel hati
Source
Type III Sum of
Squares df Mean Square F Sig.
Model 974.306a 11 88.573 107.001 .000
Perlakuan 351.139 5 70.228 84.839 .000
Ulangan 6.472 5 1.294 1.564 .207
Error 20.694 25 .828
Total 995.000 36
a. R Squared = ,979 (Adjusted R Squared = ,970)
Post Hoc Tests Perlakuan
Homogeneous Subsets
Jumlah inti sel hati
Duncan Perlakuan N Subset 1 2 3 4 T3E 6 1.3333 T3B 6 2.1667 2.1667 T3A 6 2.8333 T3C 6 2.8333 T3D 6 5.0000 T3F 6 10.6667 Sig. .125 .242 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means.
Lampiran 14 Uji statistik (Anova) dengan SPSS untuk jumlah inti sel hati tikus pada terminasi hari ke-22 dengan intensitas kandungan Cu,Zn-SOD negatif (-)
Univariate Analysis of Variance
Tests of Between-Subjects Effects
Dependent Variable: Jumlah inti sel hati
Source
Type III Sum of
Squares df Mean Square F Sig.
Model 383.667a 11 34.879 15.479 .000
Perlakuan 147.333 5 29.467 13.077 .000
Ulangan 11.333 5 2.267 1.006 .435
Error 56.333 25 2.253
Total 440.000 36
a. R Squared = ,872 (Adjusted R Squared = ,816)
Post Hoc Tests Perlakuan
Homogeneous Subsets
Jumlah inti sel hati
Duncan Perlakuan N Subset 1 2 T3A 6 1.3333 T3B 6 1.5000 T3C 6 1.5000 T3E 6 1.6667 T3D 6 2.0000 T3F 6 7.0000 Sig. .499 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means.