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Pembuatan Media Agar
Media agar buatan pabrik ditimbang sesuai dengan petunjuk yang ada pada labelnya. Agar ditimbang sesuai keperluan. Setelah itu ditambahkan air steril. Kemudian direbus dan disterilisasi dengan autoklave. Setelah itu dibiarkan hangat-hangat kuku dan tambahkan antibiotik, lalu dituangkan ke dalam cawan Petri atau tabung sesuai kebutuhan.
Lampiran 80
Prosedur Penghitungan Telur Pergram Tinja (EPG)
Prosedur dilakukan berdasarkan metode Whitlock (1948) yang telah dimodifikasi. Dilakukan sebagai berikut: Tiga gram tinja dimasukkan ke dalam botol 60 ml. Dan ditambahkan air sebanyak 17 ml, diinkubasi, lalu feses dalam botol dihomogenkan dengan cara memblendernya lalu ditambahkan larutan garam jenuh sebanyak 40 ml, kemudian sambil diaduk dengan menggunakan pipet yang ujungnya telah dilengkapi saringan, dihisap suspensi tinja dan di masukkan ke dalam ruang dari kamar hitung Universal Whitlock, didiamkan selama 1-2 menit, lalu spesimen diperiksa di bawah mikroskop dengan perbesaran rendah (4x10) dan dihitung jumlah telur cacing yang ditemukan. Hasil penghitungan telur cacing untuk setiap satu gram sampel tinja (EPG)= jumlah telur cacing yang ditemukan dikalikan 20.
Lampiran 81
Penghitungan Larva Pergram Tinja (LPG)
Penghitungan dilakukan berdasarkan Manual of Veterinary Parasitological Laboratory Techniques [MAFF] (1971) yang telah dimodifikasi sebagai berikut : Untuk pemupukan, tinja domba dan media vermikulit dihomogenkan pada mortar. Campuran tersebut dimasukkan ke dalam botol pido, lalu dipadatkan. Sisi bagian dalam botol dibersihkan dengan air, kemudian ditutup dengan tutup wadah
plastik,
lalu
diinkubasi
dan
dijaga
kelembababannya
dengan
menambahkan air secukupnya. Setelah 6 hari di inkubasi larva dipanen dengan cara mengisi air ke dalam botol tersebut sampai penuh pada bibir botol lalu ditutup dengan cawan Petri, kemudian posisi botol dibalik sehingga tutup botol (cawan Petri) berada dibawah, pada sekeliling cawan Petri diberi air. Setelah 1224 jam, cairan tersebut ditampung pada botol lain untuk dihitung jumlah larva cacing. Cairan tersebut dipusing dengan sentrifus lalu diendapkan larvanya kemudian dihitung jumlah larva. Larva pergram tinja disesuaikan dengan berat tinja yang diperiksa dan jumlah air yang digunakan.
Lampiran 82
Pembuatan Preparat Slide
Pembuat slide dilakukan menurut Drury and Wallington (1980). Pembuatan dilakukan sebagai berikut; Cacing yang telah difiksasi ke dalam BNF 10% selama 2 hari, kemudian di masukkan ke dalam kaset, lalu dilakukan proses Hidrasi pada mesin otomatis ”Sakura” selama 1 hari lalu di vakum di buat embending (dicetak bloking) pakai parafin, setelah padat di potong dengan mikrotom setebal 3-5µm, lalu dilanjutkan dengan pewarnaan yang dikehendaki.
Lampiran 83
Pembuatan Larutan Buffer Neutral Formalin (BNF)
Larutan pengawet Buffer Neutral Formalin/ Phosphade Buffered (Neutral) Formalin (BNF)
pH 7-7,2 dibuat berdasarkan Vacca (1985) Adapun cara
pembuatannya sebagai berikut : Mempersiapkan bahan- bahan sebagai berikut : 1. 38- 40% Formaldehyde
100 ml
2. Sodium Phosphatase monobasic (Na H2P04)
4 gr
3. Air steril
900 ml
4. Sodium Phosphatase di basic (anhydrous) Na2HPO4
6,5 gr
Bahan no 2 di tambahkan pada no 3 lalu tambahkan pula bahan no 4, kemudian no 1, buat pH yang diinginkan dengan menambahkan asam atau basa secukupnya.
Lampiran 84
Pewarnaan dengan Hemaktosilin Eosin (HE)
Pewarnaan HE dilakukan dengan cara merendam pada larutan Xylol 1 dan 2, masing-masing selama 3 menit, lalu direndam pada larutan etanol bertingkat absolut s/d 70%, masing-masing selama 3 menit lalu di biarkan pada air mengalir 3 menit, lalu direndam pada larutan pembiru selama 1 menit, di bilas dan dibiarkan pada air mengalir selama 3 menit, lalu rendam pada larutan Eosin selama 8 menit, kemudian di biarkan pada air mengalir selama 3 menit lalu rendam dengan etanol bertingkat 70% s/d absolut masing-masing selama 3 menit, lalu rendam dengan xylol 1:2 masing-masing selama 3 menit, lalu cover direkatkan pada slide preparat dengan Debuitril Platalat Xylol (DPX).
Lampiran 85
Pewarnaan dengan Minyak Cengkeh (MC)
Mikroba yang akan diwarnai dengan minyak cengkeh diberi perlakuan sebagai berikut ; Mikroba yang telah difiksasi dengan BNF 10% atau segar direndam dalam KOH 1% selama 10 detik, lalu direndam dalam minyak cengkeh selama 10 detik, kemudian dicuci dengan aquades. Lalu dicuci dengan alkohol 70%, lalu 95%, lalu 100%. Kemudian dimounting dengan Entellan.
Lampiran 86
Naskah Publikasi
Ahmad RZ, Satrija F, Sukarno N, Pasaribu FH. 2007. Daya reduksi cendawan Duddingtonia flagrans dan Saccharomyces cerevisiae terhadap larva cacing Haemonchus contortus pada domba. J Vet. Unud (8) 1: 46-52. Ahmad RZ, Satrija F, Sukarno N, Pasaribu F, Beriajaya. 2007. Pengaruh jumlah telur, daya tetas, larva pergram tinja, cacing Haemonchus contortus setelah pemberian Duddingtonia flagrans dan Sacharomyces cerevisiae pada domba. J. Media veteriner IPB (in press)
