Efektifitas Pemberian Soyghurt yang Mengandung Bakteri Asam Laktat dalam Memperbaiki Kerusakan Jaringan Hati Mencit (Mus musculus L.) yang Dipapar Timbal

Lampiran A. Ethical Clearance

Lampiran B. Ringkasan Kegiatan Penelitian

Pembuatan Susu Kedelai

Pembuatan Starter Soyghurt

Inokulasi

Inkubasi

Pemeriksaan Kadar SGOT/SGPT

Hewan Percobaan

C, 4 jam

Kultur

Soyghurt II

Suhu 40

C, 6 jam Soyghurt III Suhu 40 o

C, 8 jam Soyghurt I Suhu 40 o

Penghitungan Jumlah

Koloni dan

Identifikasi Bakteri

Pemeriksaan

Makroskopis Hati dan

Histologi Hati

Lampiran C. Skema Perlakuan Hewan Coba Mencit (Mus musculus L.) N= 28 ekor

Aklimatisasi selama 1 minggu Secara random dibagi dalam 4 kelompok

K P1 P2 P3

7 ekor 7 ekor 7 ekor 7 ekor

K P1 P2 P3

Aquadest Soyghurt Timbal

Timbal 0,5 ml

0,5 ml 0,5 ml 0,5 ml +

Selama 14 h Selama 14 h Soyghurt Selama 14 h 0,5 ml

Selama 14 h

Pada hari ke-15, 7 ekor mencit dari tiap-tiap

kelompok didekapitasi

Dilakukan pemeriksaan: Berat hati

Makroskopis hati (warna, konsistensi, struktur permukaan)
Kadar SGOT dan SGPT

Lampiran D. Diagram Alir Pembuatan Starter Soyghurt (Koswara, 2006) Susu bubuk 80 g + gula 2% Dilarutkan dengan air mendidih pada suhu o

90 C sebanyak 500 ml selama 30 menit

o Suhu diturunkan sampai 40-45 C Diinokulasikan dengan yoghurt komersial 4% atau kultur murni S.thermophilus dan L.bulgaricus o

Diinkubasi pada suhu 40 C selama 8 jam

STARTER SOYGHURT

Lampiran E. Diagram Alir Pembuatan Susu Kedelai (Yudhi, 2008) 1 kg kacang kedelai disortasi dan dicuci bersih

Direndam dalam air yang mengandung NaHCO 3 0,5% selama 8 jam, cuci dan ditiriskan

Kedelai dimasukkan ke dalam air mendidih dgn suhu 80-90 o

C selama 30 menit, ditiriskan dan dikupas kulitnya Diblender dengan air panas

90 o

C (jumlah air 6:1) sampai menjadi bubur Disaring, filtrat ditampung ampasnya dibuang Filtrat dipanaskan

80-90 o

C selama 30 menit menit, didinginkan sampai suhu 40 o

C SUSU KEDELAI

Lampiran F. Diagram Alir Pembuatan Soyghurt (Koswara, 2006) Susu kedelai 435 ml + susu bubuk 65 g

Ditambah gula pasir 2% Inokulasi dengan kultur starter 4% Diaduk sampai larut Dimasukan dalam wadah ditutup dengan plastik poliethilen dan dilubangi o

Diinkubasi pada suhu 40 C (4 jam, 6 jam, 8 jam) Soyghurt

Lampiran G. Reaksi Karbohidrat dari Genus Lactobacillus (Buchanan dan Gibbons, 1974) N o Jenis Karbohidrat S p e c i e s

14 Melibiose - - - - - - d + - - - - + - d

7 Glucose (gas) - - - - - - - - - - - - - - -

8 Gluconate - - - - - - - - + + + + + + +

9 Lactose - + - + + + + + # + + # + d -

10 Maltose d + + + - + + + d - + + + + +

11 Mannitol - - - - - - - + + - + + + - +

12 Mannose + + + + - + + + + + + + + + +

13 Melezitose - - - - - - - - + - + - d - -

15 Raffinose - - - - - - d + - - - - + - d

5 Galactose # # + + - + + + + + + + + + +

16 Rhamnose - - - - - - - - - - - - - - +

17 Ribose - - - - - - - - + + + + + + -

18 Salicin - + + + - - + - + + + + + + d

19 Sorbitol - - - - - - - + + + + - + - d

20 Sucrose + + + + - - + + d - + + + - +

21 Trehalose - + + + - # + + + - + + + - -

22 Xylose - - - - - - - - - - - + d - -

6 Glucose (acid) + + + + + + + + + + + + + + +

4 Fructose + + + + + # + + + + + + + + +

3 Cellobiose - + + - - - + - + + + + + + -

1 Amygladin - + + - - - + - + - + + + - -

2 Arabinose - - - - - - - - - - - - d - -

23 Esculin - + + # - - + # + - + - + + d

Keterangan: (1) L. delbrueckii, (2) L. leichmannii, (3) L. jensenii, (4) L. lactis, (5) L. bulgaricus, (6) L. helveticus, (7) L. acidophilus, (8) L. salivarius,

(9) L. casei, (10) L. casei subsp. tol erans, (11) L. casei subsp. pseudoplantarum, (12) L. xylosus, (13) L. plantarum, (14) L. curvatus, (15) L. coryniformis, (-) reaksi negatif, (+) reaksi positif, (#) reaksi lemah, (d) beberapa strain +

20 Trehalose

13 Esculine

14 Salicine

15 Manitol

16 Maltose

17 Lactose

18 Galactose

19 Gelatin

21 Cellobiose

11 Amygladin

22 Sacharose

23 Melezitose

24 Inulin

25 Raffinose

26 Gluconat

27 Adonitol

28 Indol

29 Urease

12 Arbutine

10 Rhamnose

Lampiran H. Pola Fermentasi Gula oleh Bakteri Asam Laktat (Safitri, 2010) N o Jenis Gula S p e c i e s

9 Sorbitol

1 Glycerol

2 Arabinose

3 Ribose

4 Silose

5 Glucose

6 Fructose

7 Mannose

8 Sorbose

Keterangan: (1) Lactobacillus acidophilus, (2) Lactobacillus paracasei, (3) Lactobacillus

bulgaricus , (4) Lactobacillus delbrueckii, (5) Lactobacillus brevis, (6) Pediococcus pentosaceus (7) Pediococcus damnosus (8) Pediococcus acidilactici (9) Bifidobacterium spp. (10) Actinomyces israelii, (-) reaksi negatif, (+) reaksi positif.

Lampiran I. Reaksi Karbohidrat dari Genus Streptococcus (Buchanan dan

Gibbons, 1974)

Spesies Pertumbuhan Hidrolisa

o o

Streptococcus

10 C

45 C Gelatin Pati

- - S. thermophilus

S. salivarius

S. lactis
S. cremoris

Reaksi Karbohidrat

S. thermophilus S. salivarius S. lactis S. cremoris

Glucose
Fruc>
Lac
Sucrose

Trehalose

Maltose

Inulin
Glycerol

Manitol - - Sorbitol

Salicin

/- +/-

Raffinose - +* Xylose

Arabinose + - Esculin

d d Keterangan: (-) reaksi negatif; (+) reaksi positif; (+*) jika positif reaksi lambat; (+/-) beberapa strain positif, beberapa negatif

Test of Homogeneity of Variance a,b

% ke ru sa ka n is co n sta n t w h e n K e lo m p o k = P 2 . It h a s b e e n o m itte d . b .

% kerusakan is constant w hen Kelom pok = P2. It has been om itted.

Lilliefors Significance C orrection a. % kerusakan is constant w hen Kelom pok = K. It has been om itted.

Shapiro-Wilk This is a low er bound of the true significance. *.

Kolmogorov-Smirnov

Berat Hati SGOT SGPT Statistic df Sig. Statistic df Sig.

Kelompok K P1 P2 P3 K P1 P2 P3 K P1 P2 P3

.249 7 .200 * .844 7 .108 .257 7 .179 .843 7 .106 .248 7 .200 * .924 7 .502 .223 7 .200 * .922 7 .487 .130 7 .200 * .974 7 .924 .148 7 .200 * .950 7 .729 .196 7 .200 * .944 7 .671 .149 7 .200 * .953 7 .759 .207 7 .200 * .905 7 .365 .204 7 .200 * .915 7 .433 .199 7 .200 * .910 7 .399 .229 7 .200 * .893 7 .292

Tests of Normality

b,c

% ke ru sa ka n is co n sta n t w h e n K e lo m p o k = K . It h a s b e e n o m itted . a .

1.891

Levene Statistic df1 df2 Sig.

Berat Hati SGOT SGPT

3 24 .018 Based on Mean Based on Median Based on Median and with adjusted df Based on trimmed mean Based on Mean Based on Median Based on Median and with adjusted df Based on trimmed mean Based on Mean Based on Median Based on Median and with adjusted df Based on trimmed mean

3 24 .096 2.371 3 21.036 .099 4.085

3 24 .017 2.371

3 24 .157 4.134

3 24 .226 1.557 3 19.747 .231 1.896

3 24 .162 1.557

3 24 .171 1.866

1.818

3 24 .636 .576 3 18.251 .638

3 24 .158 .576

Lampiran J. Tabel Uji Statistik

ANOVA

Sum of Squares df Mean Square F Sig. Berat Hati Between Groups

.011 3 .004 4.969 .008 Within Groups .018 24 .001 Total

.029

27 SGOT Between Groups 475064.0 3 158354.667 283.161 .000

Within Groups 13421.714 24 559.238

Total 488485.7

27 Multiple Comparisons

Bonferroni

Mean

95% Confidence Interval

Difference

Dependent Variable (I) Kelompok (J) Kelompok (I-J) Std. Error Sig. Lower Bound Upper Bound

Berat Hati K P1 .02429 .01451 .643 -.0174 .0660

.02714 .01451 .442 -.0689 .0146 P3 .01714 .01451 1.000 -.0246 .0589 P1 K -.02429 .01451 .643 -.0660 .0174 P2 -.05143* .01451 .010 -.0931 -.0097 P3 -.00714 .01451 1.000 -.0489 .0346 P2 K

.02714 .01451 .442 -.0146 .0689 P1 .05143* .01451 .010 .0097 .0931 P3 .04429* .01451 .033 .0026 .0860

P3 K -.01714 .01451 1.000 -.0589 .0246 P1 .00714 .01451 1.000 -.0346 .0489 P2

.04429* .01451 .033 -.0860 -.0026
SGOT K P1 -303.14286* 12.64050 .000 -339.4855 -266.8002
P2
12.85714 12.64050 1.000 -49.1998 23.4855 P3 -215.42857* 12.64050 .000 -251.7712 -179.0859 P1 K 303.14286* 12.64050 .000 266.8002 339.4855 P2 290.28571* 12.64050 .000 253.9431 326.6284 P3 87.71429* 12.64050 .000 51.3716 124.0569

P2 K 12.85714 12.64050 1.000 -23.4855 49.1998 P1 -290.28571* 12.64050 .000 -326.6284 -253.9431 P3 -202.57143* 12.64050 .000 -238.9141 -166.2288

P3 K 215.42857* 12.64050 .000 179.0859 251.7712 P1 -87.71429* 12.64050 .000 -124.0569 -51.3716 P2

202.57143* 12.64050 .000 166.2288 238.9141 *. The mean difference is s ignificant at the .05 level.

Kruskal-Wallis Test Ranks Kelompok N M ean Rank SG PT K

4.64 P1

25.00 P2

10.36 P3

18.00 Total

T e s t S t a t i S G P T C h i - S q u a

2 4 . 5 0 6 d f

3 A s y m p . S . 0 0 0 a . K r u s k a l W a l l i s b .

G r o u p i n g V a r i a Mann-Whitney Test Ranks

Kelompok N Mean Rank Sum of Ranks SGPT K

4.00

28.00 P1

11.00

77.00 Total

14 % k k K 7 400 2800 b

T e s t S t a t is t ic

S G P T M a n n - W h i tn e y U

. 0 0 0 W i l c o x o n W 2 8 . 0 0 0 Z

3 .1 3 0 A s y m p . S i g . ( 2 - t a . 0 0 2

a E x a c t S i g . [ 2 * ( 1 - t . 0 0 1 S i g . ) ] a .

N o t c o r r e c t e d f o r ti e s . b .

G r o u p i n g V a r i a b l e : K e l Ranks

Kelompok N Mean Rank Sum of Ranks SGPT K

4.64

32.50 P2

10.36

72.50 Total

T e s t S t a t is t ic

S G P T M a n n - W h i tn e y U 4 .5 0 0 W i l c o x o n W

3 2 . 5 0 0 Z - 2 .5 5 8 A s y m p . S i g . ( 2 - t a . 0 1 1 a

E x a c t S i g . [ 2 * ( 1 - t . 0 0 7 S i g . ) ] a .

N o t c o r r e c t e d f o r ti e s . b .

G r o u p i n g V a r i a b l e : K e l

Ranks

Kelompok N Mean Rank Sum of Ranks SGPT K

4.00

28.00 P3

11.00

77.00 Total

14 b

T e s t S t a t is t ic

S G P T M a n n - W h i tn e y U . 0 0 0 W i l c o x o n W 2 8 . 0 0 0 Z - 3 .1 3 0 A s y m p . S i g . ( 2 - t a

. 0 0 2

E x a c t S i g . [ 2 * ( 1 - t . 0 0 1

S i g . ) ] a .

N o t c o r r e c t e d f o r ti e s . b .

G r o u p i n g V a r i a b l e : K e l

Ranks

Kelompok N Mean Rank Sum of Ranks SGPT P1

11.00

77.00 P2

4.00

28.00 Total

T e s t S t a t is t ic

S G P T M a n n - W h i tn e y U

. 0 0 0 W i l c o x o n W 2 8 . 0 0 0 Z

3 .1 3 0 A s y m p . S i g . ( 2 - t a . 0 0 2

E x a c t S i g . [ 2 * ( 1 - t . 0 0 1

S i g . ) ] a .

N o t c o r r e c t e d f o r ti e s . b . G r o u p i n g V a r i a b l e : K e l

Ranks

Kelompok N Mean Rank Sum of Ranks SGPT P1

11.00

77.00 P3

4.00

28.00 Total

14 % k k P1 7 950 6650 b

T e s t S t a t is t ic

S G P T M a n n - W h i tn e y U

. 0 0 0 W i l c o x o n W 2 8 . 0 0 0 Z - 3 .1 3 0 A s y m p . S i g . ( 2 - t a

. 0 0 2

E x a c t S i g . [ 2 * ( 1 - t . 0 0 1

S i g . ) ] a .

N o t c o r r e c t e d f o r ti e s . b . G r o u p i n g V a r i a b l e : K e l

Ranks

Kelompok N Mean Rank Sum of Ranks SGPT P2

4.00

28.00 P3

11.00

77.00 Total

3 .1 3 0

28 Kelom pok K P1 P2 P3 Total derajat kerusak an

14 Kelompok K P1 Total derajat kerusakan

77.00

11.00

28.00

4.00

Mann-Whitney Test Ranks

Kruskal W allis Tes t a. Groupi ng V ariable: Kelompok b.

Chi-Square df As ymp. Si g. derajat kerusakan

3 .000

25.036

Te st S tatistics a,b

N Mean Rank

19.50

T e s t S t a t is t ic

7.50

23.50

7.50

Derajat Kerusakan Kruskal-Wallis Test Ra nks

G r o u p i n g V a r i a b l e : K e l b .

S G P T N o t c o r r e c t e d f o r ti e s . a .

M a n n - W h i tn e y U W i l c o x o n W Z A s y m p . S i g . ( 2 - t a E x a c t S i g . [ 2 * ( 1 - t S i g . ) ]

. 0 0 2 . 0 0 1

. 0 0 0 2 8 . 0 0 0

N Mean Rank Sum of Ranks

b Te st S tatistics

derajat kerusakan Mann-W hit ney U .000 W ilcox on W

28.000 Z

3. 435 As ymp. Si g. (2-tailed) .001

Ex act Sig. [2*(1-tai led .001

Sig.)] a.

Not correct ed for ties.

Groupi ng V ariable: Kelompok

Ranks

Kelompok N Mean Rank Sum of Ranks derajat kerusakan K

7.50

52.50 P2

7.50

52.50 Total

14 b Te st S tatistics

derajat kerusakan Mann-W hit ney U

24.500 W ilcox on W 52.500 Z

.000 As ymp. Si g. (2-tailed)

1.000

Ex act Sig. [2*(1-tai led 1.000

Sig.)] a.

Not correct ed for ties.

b. Groupi ng V ariable: Kelompok

Ranks

Kelompok N Mean Rank Sum of Ranks derajat kerusakan K

4.00

28.00 P3

11.00

77.00 Total

b Te st S tatistics

derajat kerusakan Mann-W hit ney U

.000 W ilcox on W

28.000 Z

3. 500 As ymp. Si g. (2-tailed)

.000

Ex act Sig. [2*(1-tai led .001

Sig.)] a. Not correct ed for ties.

Groupi ng V ariable: Kelompok

Ranks

Kelompok N Mean Rank Sum of Ranks derajat kerusakan P1

11.00

77.00 P2

4.00

28.00 Total

14 b Te st S tatistics

derajat kerusakan Mann-W hit ney U

.000 W ilcox on W

28.000 Z

3. 435 As ymp. Si g. (2-tailed)

.001

Ex act Sig. [2*(1-tai led .001

Sig.)] a. Not correct ed for ties.

b. Groupi ng V ariable: Kelompok

Ranks

Kelompok N Mean Rank Sum of Ranks derajat kerusakan P1

9.50

66.50 P3

5.50

38.50 Total

b Te st S tatistics

derajat kerusakan Mann-W hit ney U

10.500 W ilcox on W 38.500 Z

2. 082 As ymp. Si g. (2-tailed) .037

Ex act Sig. [2*(1-tai led .073

Sig.)] a.

Not correct ed for ties.

b. Groupi ng V ariable: Kelompok

Ranks

Kelompok N Mean Rank Sum of Ranks derajat kerusakan P2

4.00

28.00 P3

11.00

77.00 Total

14 b Te st S tatistics

derajat kerusakan Mann-W hit ney U

.000 W ilcox on W

28.000 Z -3. 500 As ymp. Si g. (2-tailed) .000

Ex act Sig. [2*(1-tai led .001

Sig.)] a. Not correct ed for ties.

Groupi ng V ariable: Kelompok

Lampiran K. Hasil Uji Morfologi dan Fisiologi Bakteri Asam Laktat Gambar 1. Uji Pewarnaan Gram: