Saran SIMPULAN DAN SARAN

kerusakan sel akibat denaturasi protein, sehingga sel menjadi lebih tahanresistance terhadap perlakuan panas atau peningkatan suhu.

5.2 Saran

Perlu dilakukan penelitian lebih lanjut mengenai perubahan seluler yang terjadi pada isolat lokal Cronobacter spp. yang mampu bertahan survive selama proses pengeringan semprot sehingga mengalami peningkatan ketahanan terhadap suhu rekonstitusi 50 °C. Perubahan seluler pada isolat lokal Cronobacter spp. yang mampu bertahan selama penyimpanan pada kondisi a w susu bubuk yang berbeda RH berbeda juga perlu diketahui. Perubahan-perubahan tersebut antara lain kebocoran sel yang diindikasikan dengan meningkatnya jumlah asam nukleat dan protein dalam medium pertumbuhan Cronobacter spp., kemampuan tumbuh dan bentuk kurva pertumbuhan Cronobacter spp. pada kondisi pH, suhu, dan tekanan osmosis berbeda, serta komponen solut seperti trehalosa yang diakumulasi di dalam sel Cronobacter spp. selama mengalami stres pengeringan dan kekeringan. DAFTAR PUSTAKA AOAC. 1995. Official Method of Analysis. Association of Official Analytical Chemist, Washington DC. Archer J, Jervis ET, Bird J, Gaze JE. 1998. 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Viability of Bifidobacterium Pseudocatenulatum G4 after spray-Drying and Freeze-Drying. Microbiology Insights 3:37-43. http:www.la- press.com. World Health Organization WHO, 2007. Safe preparation, storage and handling of powdered infant formula guidelines. Available at; http:www.who.intfoodsafetypublicationsmicropif2007enindex.html. Yance PH. 2001. Water stress, osmolytes and proteins. Am. Zool. 41,699 –709. Yi SM, Zhu JL, Fu LL, Li JR. 2010. Tea polyphenols inhibit Pseudomonas aeruginosa through damage to the cell membrane. International Journal of Food Microbiology. www. e lsevier.comlocateij foodmicro Lampiran 1. Tahapan Penelitian, Tujuan dan Rancangan Percobaan No Tahapan Penelitian Tujuan Faktor AnalisisPengamatan 1. Konfirmasi kultur Cronobacter spp. pada media selektif DFI dan pada media TSA Untuk mengetahui tingkat kemurnian dan sifat-sifat Cronobacter spp. yang digunakan - Morfologi - pewarnaan Gram - bentuk dan warna koloni 2. Skrining cepat ketahanan panas isolat Cronobacter spp. pada suhu rekonstitusi 50 °C selama 30 menit Untuk mendapatkan 1 isolat Cronobacter spp. yang paling tahan pada suhu rekonstitusi 50 °C selama 30 menit Jenis isolat Cronobacter spp. Log reduksi Cronobacter spp. 3. Sintas Cronobacter sp. , dalam susu skim saat rekonstitusi sebelum mengalami proses pengeringan semprot Untuk mengetahui sintas survival Cronobacter sp. dalam skim milk saat rekonstitusi sebelum mengalami proses pengeringan semprot - 1. Log reduksi Cronobacter sp. 2. Pengamatan morfologi - pewarnaan gram - bentuk koloni dan - ukuran sel 4. Pengaruh pengeringan semprot suhu inlet 160°C dan outlet 82°C terhadap sintas Cronobacter sp. Untuk mengetahui pengaruh pengeringan semprot terhadap sintas Cronobacter sp. 1. Jumlah Log Reduksi jumlah Cronobacter sp. 2. a w aktivitas air 3. Kadar air awal susu bubuk 5. Pengaruh pengeringan semprot suhu inlet 160°C dan outlet 82°C terhadap sintas Cronobacter sp. saat rekonstitusi Untuk mengetahui pengaruh pengeringan semprot terhadap sintas Cronobacter sp. saat rekonstitusi 1. Log Reduksi Cronobacter sp. 2. Pengamatan morfologi - pewarnaan gram - bentuk koloni dan - ukuran sel No Tahapan Penelitian Tujuan Faktor - faktor AnalisisPengamatan 6. Pengaruh RH penyimpanan terhadap sintas Cronobacter sp. Untuk mengetahui pengaruh RH penyimpanan terhadap sintas Cronobacter sp. 1. RH penyimpan an 50, 70, dan 90 1. Junlah Cronobacter sp. 2. a w aktivitas air 3. Kadar air awal susu bubuk 7. Pengaruh RH penyimpanan terhadap sintas Cronobacter sp. saat rekonstitusi Untuk mengetahui pengaruh RH penyimpanan terhadap sintas Cronobacter sp. saat rekonstitusi 2. RH penyimpan an 50, 70, dan 90 1. Log reduksi Cronobacter sp. 2. Pengamatan morfologi - pewarnaan gram - bentuk koloni dan - ukuran sel Lampiran 2. Hasil Konfirmasi Isolat Cronobacter spp. 1. Aktivasi kultur stok Cronobacter spp. dalam medium BHI broth 2. Pewarnaan gram Mikroskop Cahaya Perbesaran 1000x 3. Konfirmasi 8 isolat Cronobacter spp. dalam media selektif DFI Agar 4. Konfirmasi 8 isolat Cronobacter spp. dalam media TSA Agar Lampiran 3. Proses Pengeringan Semprot dengan Spray Drier Suhu Inlet ±160 °C Suhu Outlet ±82 °C Susu skim hasil pengeringan semprot suhu inlet 160°C dan outlet 82°C a b Bentuk koloni Cronobacter spp. YRc3a dalam media TSA-YE sebelum a dan sesudah pengeringan semprot b Lampiran 4. Hasil Perhitungan Sintas Cronobacter spp. YRc3a Selama Proses Pengeringan Semprot 1. Jumlah inokulum hasil sentrifugasi ± 10 11 cfuml 3000 rpm15 menit Ulangan Jumlah Inokulum biomasa sel Pengenceran ∑ koloni CFUml I -8 TBUDTBUD 4.05x10 11 3.46 x 10 11 -9 9189 -10 48 II -9 224188 2.41 x10 11 -10 6751 -11 105 2. Reduksi jumlah sel setelah mengalami proses pengeringan semprot jumlah awal sel merupakan jumlah sel awalbiomasa sel. a. Jumlah awal sel yang diinokulasikan ± 2 ml biomasa sel dalam BPW dimana: - Jumlah selml 3.46 x 10 11 cfuml Total jumlah sel yang diinokulasikan 6.92 x 10 11 cfu 2 ml - Jumlah sel setelah diinokulasikan dalam 450 ml 180 gram270 ml dalam ml aktualnya pada tabel berikut : Ulangan Jumlah Sel dalam 450 ml skim milk Rata-rata CFUml Pengenceran ∑ koloni CFUml I -7 2922 2.55x10 8 2.2 x10 8 -8 47 -9 10 II -7 2314 1.85 x10 8 -8 51 -9 10 - Jumlah sel dalam gram skim milk 180 gram = 3,84 x 10 9 CFUg b. Hasil pengeringan semprot sebanyak ± 45 gram hasil analisa survival sel dalam skim milk yang direkonstitusi 2.2 gram 15 ml adalah : Ula Ngan Survivor 27 o

C: ± 1menit Rata-

rata CFU ml ∑ koloni Survivor 50 o

C: ± 1menit Rata-

rata CFU ml Pengen Ceran ∑ koloni Pengen ceran Pengen ceran ∑ koloni CFU ml I -3 3515 2.5 x10