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Microbiology

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Laporan Praktikum Tanggal Praktikum: Kamis, 22 Maret 2012
Mikrobiologi Pangan PJ Praktikum : Lukie Trianawati, STP, Msi. Asisten : Mariska Pratami P, A.Md

PENGGUNAAN MIKROSKOP
(Pengamatan Morfologi Kapang dan Khamir Menggunakan Mikroskop)
Kelompok 2/BP1
Putra Mahardiman J3E111078
Dian Mawarni J3E111027
Dina Crownia J3E111087
Niken Indah C. J3E111081
Rosi Rizki R. J3E111034

SUPERVISOR JAMINAN MUTU PANGAN
PROGRAM DIPLOMA
INSTITUT PERTANIAN BOGOR
2012

Mikroorganisme atau mikroba adalah organisme yang berukuran sangat kecil sehingga untuk mengamatinya diperlukan alat bantuan. Mikroorganisme disebut juga organisme mikroskopik. Mikroorganisme seringkali bersel tunggal (uniseluler) maupun bersel banyak (multiseluler). Namun, beberapa protista bersel tunggal masih terlihat oleh mata telanjang dan ada beberapa spesies multisel tidak terlihat matatelanjang. Virus juga termasuk ke dalam mikroorganisme meskipun tidak bersifat seluler.
Pengamatan bentuk, ukuran, dan identifikasi jenis bakteri akan tampak jelas jika dilakukan pewarnaan terhadap sel. Metode pewarnaan sel bakteri pertama kali dikembangkan oleh Christian Gram pada tahun 1994. Bakteri dapat digolongkan menjadi dua kelompok yaitu gram positif dan gram negatif didasarkan pada perbedaan struktur dinding sel. Pada umumnya bakteri gram negatif lebih tahan terhadap aktivitas antimikroba dibandingkan dengan bakteri gram positif. Perbedaan daya tahan ini disebabkan karena perbedaan komponen penyusun dinding sel (Rahayu, 2000).

Menurut Waluyo (2008), tujuan dari pewarnaan adalah untuk memudahkan melihat mikroba dengan mikroskop, memperjelas ukuran dan bentuk mikroba melihat struktur luar dan struktur dalam bakteri seperti dinding sel dan vakuola, menghasilkan sifat-sifat fisik dan kimia khas dari bakteri dengan zat warna.
Bakteri gram positif seperti Staphylococcus aureus (bakteri patogen yang umum pada manusia) hanya mempunyai membran plasma tunggal yang dikelilingi dinding sel tebal berupa peptidoglikan. Sekitar 90 persen dari dinding sel tersebut tersusun atas peptidoglikan sedangkan sisanya berupa molekul lain bernama asam teikhoat. Di sisi lain, bakteri gram negatif (seperti E. coli) memiliki sistem membran ganda di mana membran pasmanya diselimuti oleh membran luar permeabel. Bakteri ini mempunyai dinding sel tebal berupa peptidoglikan, yang terletak di antara membran dalam dan membran luarnya. Dalam proses ini, olesan bakteri yang sudah terfiksasi dikenai larutan-larutan berikut : zat pewarna kristal violet, larutan yodium, larutan alkohol (bahan pemucat), dan zat pewarna tandingannya berupa zat warna safranin. Bakteri Gram positif akan mempertahankan zat pewarna kristal violet dan karenanya akan tampak berwarna ungu tua di bawah mikroskop. Adapun bakteri gram negatif akan kehilangan zat pewarna kristal violet setelah dicuci dengan alkohol, dan sewaktu diberi zat pewarna tandingannya yaitu dengan zat pewarna air fuchsin atau safranin akan tampak berwarna merah. Perbedaan warna ini disebabkan oleh perbedaan dalam struktur kimiawi dinding selnya.
Pewarnaan sederhana merupakan teknik pewarnaan yang paling banyak digunakan. Disebut sederhana karena hanya menggunakan satu jenis zat warna untuk mewarnai organisme tersebut. Kebanyakan bakteri mudah bereaksi dengan pewarnaan-pewarnaan sederhana karena sitoplasamanya bersifat basofilik (suka dan basa). Zat-zat warna yang digunakan untuk pewarnaan sederhana umumnya bersifat alkolin. Dengan pewarnaan sederhana dapat mengetahui bentuk dan rangkaian sel-sel bakteri. Pewarna basa yang biasa digunakan untuk pewarnaan sederhana ialah memilen biru, krisdal violet dan karbol fuehsin. Pada praktikum pewarnaan gram ini alam praktikum pewarnaan gram alat dan bahan yang digunakan yaitu mikroskop, objek glass, kawat ose, bunsen, pipet tetes, kultur bakteri pada tabung kultur ( diambil pertumbuhan koloni yang paling baik ), kristal violet, iodine, alkohol 95%, dan safranin. Prinsip dari pewarnaan gram yaitu pewarnaan, pencucian, dan pembandingan.
Spora bakteri adalah bentuk bakteri yang sedang dalam usaha mengamankan diri terhadap pengaruh buruk dari luar (Buckle,1985). Setelah keadaan luar membaik, maka kotak spora akan terpecah sehingga bakteri dapat tumbuh. Pelczar (1986), menyatakan bahwa spora merupakan tubuh bakteri yang secara metabolik mengalami dormansi, dihasilkan pada fase lanjut dalam pertumbuhan sel bakteri yang sama seperti asalnya, yaitu sel vegetatif. Spora umumnya disebut endospora, yaitu spora yang dibentuk di dalam sel. Endospora jauh lebih tahan terhadap pengaruh luar yang buruk dari pada bakteri biasa seperti bakteri dalam bentuk vegetatif (Musyaffalab,2010). Kondisi sporulasi dapat dicegah, jika selalu terjadi pemindahan ke medium baru. Endospora yang dibuat irisan dapat memperlihatkan pembungkus luar, korteks dan inti yang mengandung struktur nukleus. Apabila sel vegetatif membentuk endospora, sel ini membuat enzim baru, memproduksi dinding sel yang sama sekali baru dan berubah bentuk. Dengan kata lain sporulasi adalah bentuk sederhana diferensiasi sel dan diteliti secara mendalam untuk mempelajari peristiwa apa yang memicu perubahan enzim dan morfologi (Musyaffalab,2010). Menurut Volk & Wheeler (1988), dalam pengamatan spora bakteri diperlukan pewarnaan tertentu yang dapat menembus dinding tebal spora. Contoh dari pewarnanya adalah dengan penggunaan larutan hijau malakit 5%, dan untuk memperjelas pengamatan, sel vegetative juga diwarnai dengan larutan safranin 0,5% sehingga selnya berwarna merah. Dengan demikian ada atau tidaknya spora dapat teramati, bahkan posisi spora di dalam tubuh sel vegetative juga dapat diidentifikasi. Namun ada juga zat warna khusus untuk mewarnai spora dan dalam proses pewarnaannya melibatkan treatment pemanasan, yaitu; spora dipanaskan bersamaan dengan zat warna tersebut sehingga memudahkan zat warna tersebut untuk meresap ke dalam dinding pelindung spora bakteri.
Identifikasi isolat fungi dilakukan melalui dua tahap. Tahap pertama yaitu, pengamatan fungi secara makroskopis yang meliputi pengamatan terhadap warna dan bentuk koloni. Tahap kedua yaitu, pengamatan secara mikroskopis yang dilakukan dengan membuat slide kutur yang meliputi pengamatan terhadap bentuk hifa dan ukuran konidia. http://file.upi.edu/Direktori/FPMIPA/JUR._PEND._BIOLOGI/196611031991012-YANTI_HAMDIYATI/cara_membuat_slide_culture.pdfGambar 3.1 Tahap pembuatan slide kultur : (A) Potongan agar yang diambil dari medium PDA. (B) Cawan Petri berisi batang penahan dan gelas objek. (C) Inokulasi fungi pada agar yang disimpan di atas gelas objek.
(D) Agar yang telah diinokulasi ditutup dengan kaca penutup. (Sumber : www.botany.utoronto.ca) Fungi adalah nama regnum dari sekelompok besar makhluk hidup eukariotikheterotrof yang mencerna makanannya di luar tubuh lalu menyerap molekul nutrisi ke dalam sel-selnya. Fungi memiliki bermacam-macam bentuk. Awam mengenal sebagian besar anggota Fungi sebagai jamur, kapang, khamir, atau ragi, meskipun seringkali yang dimaksud adalah penampilan luar yang tampak, bukan spesiesnya sendiri. Kesulitan dalam mengenal fungi sedikit banyak disebabkan adanyapergiliran keturunan yang memiliki penampilan yang sama sekali berbeda (ingatmetamorfosis pada serangga atau katak). Fungi memperbanyak diri secaraseksual dan aseksual. Perbanyakan seksual dengan cara :dua hifa dari jamur berbeda melebur lalu membentuk zigot lalu zigot tumbuh menjadi tubuh buah, sedangkan perbanyakan aseksual dengan cara membentuk spora, bertunas atau fragmentasi hifa. Jamur memiliki kotak spora yang disebut sporangium. Di dalam sporangium terdapat spora.

http://id.wikipedia.org/wiki/Mikroorganisme

Waluyo L. 2008. Teknik dan Metode Dasar dalam Mikrobiologi. Universitas Muhammadiyah Malang Press. Malang.

http://makalahdanskripsi.blogspot.com/2008/08/pewarnaan.html

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