DAFTAR GAMBAR
SIMPULAN DAN SARAN Simpulan
Simpulan
- Diperoleh 72 isolat hasil solasi bakteri asal saluran pencernaa ayam broiler yang tidak diberi antibiotik, terdiri dari 38 isolat yang tumbuh pada pH 7.0 dan 34 isolat yang tumbuh pada pH 4.5.
- Empat isolat terpilih (7n, 25n, 27n, 34n) memiliki aktivitas penghambatan cukup bagus terhadap EPEC K1-1, E .coli, Salmonella enteric, Salmonella
subsp.2 asal ayam. Dimana Isolat 7n diperoleh pada bagianduodenum. isolat 25n, 27n, 34n pada bagianintestinum crassum.
- Hasil identifikasi keempat isolat termasuk kelompokBacillus.
- Aktivitas penghambatan tertinggi terhadap EPEC K1-1 oleh isolat 7n (9mm) pada media MRS modifikasi dengan suhu dan pH inkubasi 500C dan pH 7.0.
- Penghambatn tertinggi terhadap E. colioleh isolat 25n (29 mm) pada media MRS modifikasi,dengan suhu dan pH inkubasi 400C pada pH 8.0.
- Aktivitas penghambatan terhadap Salmonella enteric oleh isolat 7n pada media MRS modifikasi pada suhu dan pH ingkubasi 300C dan pH 5.0. - Penghambatan terhadap Salmonella subsp.2 asal ayam oleh isolat 34n
sebesar 19mm pada media MRS modifikasi suhu 370C dengan pH 9.0. - Isolat 7n, 25n, dan 34n menghasilkan enzim amilase, protease, lipase,
selulase ekstraseluler. Isolat 27n meghasilkan ketiga enzim kecuali lipase. - Isolat 7n mempunyai nilai indeks paling tinggi dengan indeks amilase 0.67,
indeks protease 1.5, indeks lipase 1,dan indeks selulase 2.
Saran
- Perlu dilakukan penelitian lebih lanjut untuk mengetahui aktivitas enzim ekstraseluler secara kuantitatif dari keempat isolat dan karakterisasi senyawa antibakteri yang dihasilkannya.
- Penelitian lanjutan secara in vivo kemampuan isolat menghambat mikrob patogen dan konversi pakan yang paling tepat untuk memperoleh efektivitas penghambatan terhadap EPEC K1-1, E. coli asal ayam, Salmonella enteric,
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68
Lampiran 1 Komposisi kimia molase
No Komponen Kisaran %) Rata-ata
1 Air 17 – 25 20 2 Sukrosa 30 – 40 35 3 Glukosa 4 – 9 7 4 Fruktosa 5 –12 9 5 Gula pereduksi 1 – 5 3 6 Karbohidrat lain 2 – 5 4 7 Abu 7 – 15 12 8 Komponen nitrogen 2 – 6 4.5
9 Asam bukan nitrogen 2 – 6 5
10 Lilin, steroiddanfosfolipid 0.1– 1 0.4
Lampiran 2 Komposisi kimia tepung kedelai
No Komponen Kadar komponen(%)
1 Protein 42.59 2 Nitrogen 6.81 3 Air 4.64 4 Lemak 19.74 5 Magnesium 0.15 6 Mangan 0.16 7 Besi 0.05 8 Seng 0.15 9 Fosfor 1.48 10 Kalsium 0.45
Lampiran 3 Komposisi media peremajaan, produksi, serta uji daya hambat isolat asal saluran pencernaan ayam broiler terhadap EPEC K1-1, E. coli
asal ayam,Salmonella enteric,Salmonellasp. asal ayam
No Nama media Komposisi Jumlah
(g/l)
1 Nutrient agar(NA) Beef extract 3
Bacto peptone 5
Bacto agar 15
2 Nutrient agar(NA) 50% semipadat Beef extract
Bacto peptone
3 5
Bacto agar 9
3 Nutrent Broth (NB) Beef extract 3
Bacto peptone 5
4 Trypticase Soy Agar(TSA) Pangkreatic digest of
casein Enzymatic digest os soybean meal 17 3 Dextrosa 2.5 Sodium chlorida 5 Dipotasium phosphate 2.5 Bacto agar 10
5 Trypticase Soy Broth(TSB) Pangkreatic digest of
casein Enzymatic digest of soybean meal 17 3 Dextrosa 2.5 Sodium chlorida 5 Dipotasium phosphate 2.5
6 Tripton Glucosa Yeas ekstract
(TGY)
Pangkreatic digest of casein
5 2.5
70
Yeast ekstract
Dekstrose 1
7 De Mann Rogosa Sharpe(MRS) Glukose 20
Pepton casein 10 Beef ekstract 8 Natrium acetate 3H2O 5 Yeast ekstract 4 K2HPO4 2 Triamonium sitrat 0.2 MgSO47H2O 0.2 MnSO44H2O 0.05 Sorbitan monooleat / Tween 80 1
8 TGY modifikasi Soybean meal 5
Yeast ekstract 2.5
Molase 1
9 MRS modifikasi Molase 20
Soy bean meal 10
Beef ekstract 8 Natrium acetate 3H2O 5 Yeast ekstract 4 TSP 2 Urea 0.2 MgSO47H2O 0.2 MnSO44H2O 0.05
Lampiran 4 Komposisi Pereaksi Pewarnaan
No. Nama media Komposisi Jumlah (g/l)
1 Pewarnaan Gram
A.Ungu kristal(Hucker’s))
Larutan A Ungu kristal(90%) 2.0 g
Etil alkohol(95%) 20.0 ml
Larutan B Amonium oksalat 0.8 g
Aquades 80.0 ml
B.Iodium Gram Iodium 1.0 g
Kalium iodida( KI) 2.0 g
Aquades 300.0 ml
C.Etil alkohol95% Etil alkohol100% 95.0 ml
Aquades 5.0 ml
D.Safranin SafraninO 0.25ml
Etil alkohol95% 10.0 ml
Aquades 100.0 ml
2 Pewarnaan spora
A.MalakitHijau Malakithijau 50.0 g
Aquades 100.0 ml
B.Safranin SafraninO 0.25ml
Etil alkohol95% 10.0 ml
72
Lampiran 5 Hasil isolasi dan identifikasi bakteri asal saluran pencernaan ayam
broilerpada media NA pH 7
Bakteri target Kode Isolat Warna koloni Karakteristik koloni Pewarnaan Gram Bentuk sel dan penataan Endo spora E. coli S. enteric Ephec K1-1 pH 7 PN.I.3.6 (1) putih b, lc, d tu tu 0 0 0 PN.I.3.2 (2) putih b, ld, c tu tu 0 0 0 PN.I.3.4 (3) putih b, lc, d tu tu 0 0 0 PN.I.3.3 (4) putih b, ld, c tu tu 0 0 0 PN.I.3.1 (5) putih b, ld, c tu tu 0 0 0 PN.II.2.1 (6) putih b, lc, c tu tu 0 0 0
PN.II.3.1 (7) putih b, ld, c positif steptobasil 1 0 0 PN.II.2.2 (8) putih b, lc, d positif basil 1 0 0
PN.II.3.3 (9) putih tb, lc, d tu tu 1 0 0 PN.II.3.2 (10) putih b, ld, c tu tu 1 0 0 PN.I.3.5 (11) putih b, d, lc tu tu 0 0 0 PN.III.3.1 (12) putih b, lc, c tu tu 1 0 0 PN.III.2.3 (13) putih b, lc, c tu tu 1 0 0 PN.III.3.3 (14) putih b, lc, d tu tu 0 0 0
PN.III.3.4 (15) putih b, lc, c negatif bacilus 1 1 0
PN.I.2.4 (16) bening tb, lc, c tu tu 0 1 0
PN.III (17) krem b, lc, c negatif cocus 1 1 1
PN.III.3.1 (18) Krem sk, lc, bt tu tu 0 0 1 PN.III (19) putih t, tb, tb tu tu 1 0 1 AN.I.2.2 (20) putih b, lc, c tu tu 1 0 1 PN.III.3.5 (21) putih t, lc, b tu tu 1 1 1 PN.III (22) putih t, lc, b tu tu 1 0 0 PN.III (23) putih st, lc, b tu tu 0 0 1 PN.III (24) putih t, lc, b tu tu 1 0 1
PN.III.2.2 (25) putih c, lc, b positif bacilus 1 1 1
PN.III (26) putih st, lc, b tu tu 1 0 1
PN.III (27) putih sk, bo, bt positif basil kecil 1 1 1
PN.III (28) putih t, lc, b negatif cocus 0 0 1
AN.II.3.3 (29) putih t, lc, b tu tu 0 0 1
AN.III.3.3 (30) putih d, bo, tb tu tu 0 0 1
AN.III.3.1 (31) putih d, bo, d tu tu 0 0 1
AN.III.2.3 (32) putih d, lk, tb tu tu 0 0 0
AN.III.1.1(33)a putih c, lc, b tu tu 0 0 1
AN.III.2.2 (34) putih c, lc, b positif bacilus 0 0 1
AN.III.3.1(35) krem tua c, lc, b tu tu 0 0 1
PN.III (36) putih c, lc, b tu tu 0 0 0
PN.III (37) putih c, lc, b tu tu 0 1 0
PN.III (38) krem t, lc, b tu tu 1 0 0
*PN=pengenceran menambahkan pepton pH 7, AA=pengenceran tanpa penambahan pepton, pH 7, I=duodenum, II=ileum, III=intestinum crasum b = bulat; tb = tidak beraturan; lk = berlekuk; lc = licin; ld = berlendir; d = datar; c = cembung; bk = berbukit; sk = seperti kawah; t = timbul; st = seperti tombol; bo = berombak; bl = bentuk L; bt = bundar dengan tepian timbul; k = keriput, tu=tidak diuji/tidak diamati 0=tidak ada aktivitas penghambatan, 1=ada aktivitas penghambatan
Lampiran 6 Hasil isolasi dan karakterisasi bakteri asal saluran pencernaan ayam broiler yang ditumbuhkan pada pH 4.5
Bakteri target Kode Isolat Warna koloni Karakteristik Koloni Pewarnaan Gram Bentuk sel dan penataan Endo spora E. coli S. enteric Ephec K1-1 Media pH4.5
PA.III.1.1 (1a) putih b, lk, bk tu tu 0 0 0
PA.III.3.1 (2a)) putih t, bo, bt negatif diplococus 0 1 0
PA.II.3.3(3a)2 putih t, lc, b tu tu 0 0 0
PA.III.4 (4a)) putih t, lc, b negatif cocus 0 1 0 PA.III.1.1 (5a) krem t, lc, b negatif cocus 1 1 0
PA.II.3.2(6a) krem d, bo, bl tu tu 1 0 0
PA.III.1.2 (7a) putih d, bo, bl negatif cocus 1 1 0