Penaeus monodon Fab - LPPM UB

LAPORAN PENELITIAN
HIBAH PENELITIAN STRATEGIS NASIONAL
TAHUN ANGGARAN 2009
PERAN ADHESIN PILI DAN OMP Vibrio alginolyticus
SEBAGAI IMUNOSTIMULAN PADA BUDIDAYA
UDANG WINDU (Penaeus monodon Fabr.)
Ketua
:
Dr. Ir. Maftuch, M.Si
Anggota
:
1. Ir. Ellana Sanusi, MP
2. Rachmi Nurdiani, S.Pi, MSc.
Dibiayai Oleh Direktorat Jenderal Pendidikan Tinggi, Departemen Pendidikan
Nasional, melalui DIPA Universitas Brawijaya Nomor : 0174.0/02304.2/XV/2009, tanggal
31 Desember 2008 dan berdasarkan SK Rektor Nomor : 160/SK/2009, tanggal 7
Mei 2009
UNIVERSITAS BRAWIJAYA
NOPEMBER 2009
RINGKASAN
Usul Hibah Penelitian Strategis Nasional. Peran Adhesin pili dan Omp Vibrio alginolyticus
Sebagai Imunostimulan Pada Budidaya Udang Windu (Penaeus monodon Fab.); Ketua : Dr.
Ir. Maftuch, M.Si, Anggota : Ir. Ellana Sanoesi, MP dan Rachmi Nurdiani, SPi, MSc.
Udang yang terserang penyakit pada kenyataannya masih mampu untuk
bertahan hidup dan udang yang mampu melewati fase akut akan mengalami fase
kronis. Pada fase ini, udang mampu hidup dan tumbuh relatif normal, namun
udang tersebut merupakan pembawa (carrier) yang dapat ditularkan ke udang
lain yang sehat. Terdapat tiga tingkatan pathogenik udang yang terserang
penyakit yaitu resisten, toleran dan rentan. Penelusuran tentang kemampuan
pertahanan udang pasca serangan penyakit, perlu diketahui baik dari segi
imunitas dan genetiknya. Secara imunitas, udang memiliki system imun non
spesifik (innate), yang terutama dijalankan hemosit (Sel granular dan
semigranular) berfungsi menyimpan protein antibakteri dan enzim protease tidak
aktif (inactive serine proteinase /proppA) dan dapat diaktifkan menjadi
Phenoloksidase (PO) yang berperan dalam sistem pertahanan tubuh udang. Untuk
mengaktifkan dibutuhkan bahan yang dapat merangsang system imun
(imunostimulan). Bahan imunostimulan ini banyak ditemukan di permukaan
bakteri dalam bentuk beta glukan, peptidoglikan dan LPS. Oleh karena itu
digunakan molekul adhesi pili dan OMP (molekul permukaan) V. alginolyticus
yang sudah dibuktikan oleh Maftuch, dkk., (2004) dapat membangkitkan respon
imun ikan kerapu tikus. Secara genetic serangan penyakit seringkali menyebabkan
mutasi gen DNA mitokondria. Mutasi disebabkan oleh antigen dan makromolekul
ROS yang dihasilkan tubuh sebagai bentuk respon imun terhadap antigen. Oleh
karena itu pada penelitian ini sangatlah penting untuk diketahu variasi genetic udang
pasca perlakuan imunostimulan dan infeksi.
Hasil penelitian berdasarkan uji ANOVA menunjukkan bahwa terdapat
perbedaan yang nyata pada jumlah hemosit, kandungan anion superoksida dan
aktivitas enzim protease pasca pemberian injeksi pili dan OMP (p<0,05). Uji
BNT (Beda Nyata Terkecil) terhadap jumlah hemosit menunjukkan bahwa dosis
2 ppt memiliki nilai rerata tertinggi yaitu 1.675.000 sel/ml. Uji BNT pada
kandungan anion superoksida menunjukkan bahwa dosis 2 ppt memiliki nila
rerata absorbansi yang tertinggi yaitu 0,07333. Uji BNT terhadap aktivitas
enzim protease menunjukkan bahwa dosis 2 ppt memiliki nilai rerata tertinggi
yaitu 18,988 unit. Hasil uji hemaglutinasi keempat berat molekul protein Omp
didapatkan bahwa pita protein Omp 42,95 kDa menunjukkan titer
hemaglutininasi tertinggi (1/32) dan untuk hasil uji hemaglutinasi keenam berat
molekul protein pili didapatkan bahwa pita protein Omp 9,08 kDa menunjukkan
titer hemaglutininasi tertinggi (1/16). Nilai phagositosis udang windu yang
bersifat SPF lebih besar dari yang lainnya. Aktifitas enzim phenoloxidase
udang windu 24 jam infeksi vibrio harveyi berbeda nyata dengan 3 jam pasca
pemberian imunostimulan. Analisis DNA mitokondria udang pasca pemberian
molekul adhesi pili dan OMP lebih baik daripada pasca serangan penyakit
SUMMARY
Proposal National Strategic Research Grant. Role of Adhesin pili and Omp Vibrio
alginolyticus For Shrimp Farming Imunostimulan In Windu (Penaeus monodon
Fab.); Chairman: Dr. Ir. Maftuch, M. Si, Members: Ir. Ellana Sanoesi, MP and
Rachmi Nurdiani, SPi, MSc.
Shrimp disease in fact still able to survive and shrimp are able to pass the
acute phase will experience a chronic phase. In this phase, the shrimp can live
and grow relatively normally, but the shrimp is a carrier (carrier) that can be
transmitted to other healthy shrimp. There are three levels of shrimp pathogenik
of disease resistant, tolerant and susceptible. Search about post-defense
capability of shrimp disease, please note both in terms of immunity and genetics.
In immunity, shrimp has a non-specific immune system (innate), which mainly run
hemosit (cells and granular semigranular) function stores antibacterial proteins
and enzymes protease inactive (inactive Serine proteinase / proppA) and can be
activated to Phenoloksidase (PO) is responsible for shrimp immune system. To
activate the required materials that can stimulate the immune system
(imunostimulan). Imunostimulan material is usually found on the surface of the
bacteria in the form of beta glukan, peptidoglycan and LPS. Therefore use pili
adhesion molecules and OMP (molecular surface) V. alginolyticus which was
proved by Maftuch, et al., (2004) can generate an immune response grouper
mice. In genetic disease often causes gene mutations of mitochondrial DNA.
Mutations caused by ROS antigens and the resulting macromolecules body as a
form of immune response against the antigen. Therefore, in this study is
important for genetic variation diketahu shrimp had given of immunostimulant and
infection.
The results based on ANOVA test showed that amount of total haemocyte
count, superoxide anion and protease activity were significantly different
(p<0,05). According to the BNT test result on total haemocyte counts showed
that 2 ppt is the optimum concentration with average 1.6750 cells/ml. The BNT
test result on superoxide anion showed that 2 ppt is the optimum concentration
with average absorbance 0,07333. The BNT test result on protease activity
showed that 2 ppt is the optimum concentration with average 18,988 units. The
results of the fourth test hemaglutinasi protein molecular weight Omp protein
bands was found that 42.95 kDa Omp showed the highest titer hemaglutininasi (1
/ 32) and for the six test results hemaglutinasi pili protein molecular weight bands
was found that 9.08 kDa Omp protein showed the highest titer hemaglutininasi (
1 / 16). Monodon shrimp phagosytosis value of SPF is greater than the others.
Phenoloxidase enzyme activity on the monodon shrimp 24 hours of Vibrio
harveyi infection had given of immunostimulant for 3 hours is significantly
different . Mitochondria DNA analysis of shrimp had given of immunostimulant
pili and adhesion molecules better than OMP post infection.
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