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Enhancement of non-specific immune response, resistance and growth of (Litopenaeus vannamei) by oral administration of nucleotide

January 2011
Jurnal Akuakultur Indonesia 10(1)

January 2011
10(1)

DOI:10.19027/jai.10.1-7

License
CC BY-SA 4.0

Authors:

Sukenda Sukenda

Bogor Agricultural University

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This research evaluated the nonspecific immune responsse, resistance, and growth of Litopenaeus vannamei fed nucleotide diet. Shrimp juveniles (mean weight 5.39±0.56 g) were reared in two groups of glass aquaria, each with three replications. Shrimps in group one and group two were fed nucleotide diet and basal diet each for four weeks. Total haemocyte count (THC) and PO activity were evaluated at the end of feeding while growth was measured at two weeks interval. At the end of feeding trial, the shrimps were intramuscularly injected with Vibrio harveyi 0.1x106 cfu.shrimp-1. THC of shrimp fed nucleotide diet significantly increased (P

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Henky Manoppo et al. / Jurnal Akuakultur Indonesia 10 (1), 1–7 (2011)

Peningkatan respons imun non-spesifik, resistensi, dan pertumbuhan udang

vaname (Litopenaeus vannamei) melalui pemberian pakan nukleotida

Enhancement of non-specific immune response, resistance and growth of

(Litopenaeus vannamei) by oral administration of nucleotide

Henky Manoppo1,2, Sukenda3, Daniel Djokosetiyanto3,

Mochamad Fatuchri Sukadi4, Enang Harris3

1 Program Doktoral Ilmu Akuakultur, Departemen Budidaya Perairan, FPIK, Institut Pertanian Bogor

2 Program Studi Budidaya, Fakultas Perikanan dan Ilmu Kelautan Universitas Sam Ratulangi, Manado

3 Departemen Budidaya, Fakultas Perikanan dan Ilmu Kelautan, Institut Pertanian Bogor

4 Pusat Riset Perikanan Budidaya KKP RI, Jakarta

ABSTRACT

This research evaluated the nonspecific immune responsse, resistance, and growth of Litopenaeus vannamei

fed nucleotide diet. Shrimp juveniles (mean weight 5.39±0.56 g) were reared in two groups of glass aquaria,

each with three replications. Shrimps in group one and group two were fed nucleotide diet and basal diet each

for four weeks. Total haemocyte count (THC) and PO activity were evaluated at the end of feeding while

growth was measured at two weeks interval. At the end of feeding trial, the shrimps were intramuscularly

injected with Vibrio harveyi 0.1x106 cfu.shrimp-1. THC of shrimp fed nucleotide diet significantly increased

(P<0.01) up to 87% higher than shrimps fed basal diet. PO activity also different significantly as compared to

shrimp fed basal diet (P<0.02) 14 days post-challenge, shrimp fed nucleotide diet showed higher resistance

(P<0.01). After 4 weeks of feeding, weight gain achieved 65.38% greater than shrimp fed basal diet (P<0.01).

As conclusion, oral administration of nucleotide at 400 mg.kg-1 diet showed positive effect on the

enhancement of nonspecific immune responsse, resistance, and growth of L. vannamei.

Key words: Litopenaeus vannamei, nucleotide, THC, PO activity, resistance

ABSTRAK

Penelitian bertujuan untuk mengevaluasi respons imun non-spesifik dan resistensi udang vaname (Litopenaeus

vannamei) yang diberi pakan nukleotida. Juvenil (5,39±0,56 g) dipelihara dalam dua kelompok akuarium kaca

masing-masing dengan 3 ulangan. Udang dalam dalam kelompok pertama diberi pakan nukleotida sedangkan

udang dalam kelompok kedua diberi pakan standar selama 4 minggu. Total haemocyte count (THC) dan

aktivitas phenoloxidase (PO) diukur pada akhir pemberian pakan sedangkan pertumbuhan udang diukur setiap

dua minggu. Pada akhir periode pemberian pakan perlakuan, udang diuji tantang secara injeksi intramuskular

dengan bakteri Vibrio harveyi 0,1x106 cfu.udang-1. THC udang yang diberi pakan nukleotida meningkat secara

signifikan (P<0,01) mencapai 87% lebih tinggi dari udang yang diberi pakan standar. Aktivitas PO udang

yang diberi pakan nukleotida juga berbeda nyata dibandingkan dengan udang yang hanya diberi pakan standar

(P<0,02). Empat belas hari setelah uji tantang, udang yang diberi pakan nukleotida memiliki resistensi yang

lebih tinggi (P<0,01). Setelah 4 minggu pemberian pakan, perolehan berat mencapai 65,38% lebih besar

(P<0,01) dibandingkan dengan udang yang hanya diberi pakan standar. Sebagai kesimpulan, pemberian secara

oral nukleotida pada level 400 mg.kg-1 pakan selama 4 minggu memberi pengaruh positif terhadap

peningkatan respons imun non-spesifik, resistensi dan pertumbuhan udang vaname.

Kata kunci: Litopenaeus vannamei, nukleotida, THC, aktivitas PO, resistensi

PENDAHULUAN

Budidaya udang telah mendapat perhatian

dunia sebab secara nyata berkontribusi dalam

perkembangan ekonomi banyak negara.

Sekalipun demikian, banyak negara-negara

produsen dihadapkan dengan masalah

munculnya penyakit secara berulang yang

mempengaruhi spesies yang dipelihara, dan

karenanya menekan kesinambungan akua-

kultur. Perkembangan penyakit bukan hanya

disebabkan oleh adanya intensifikasi pro-

Jurnal Akuakultur Indonesia 10 (1), 1–7 (2011)

Available : http://journal.ipb.ac.id/index.php/jai

http://jurnalakuakulturindonesia.ipb.ac.id

Henky Manoppo et al. / Jurnal Akuakultur Indonesia 10 (1), 1–7 (2011)

duksi tetapi juga oleh kerusakan lingkungan,

polusi, dan ketidakseimbangan nutrisi

(Bachere, 2003). Dalam dua dekade terakhir,

banyak petani atau industri budidaya udang

yang mengalami kerugian ekonomi yang

signifikan terutama disebabkan oleh penyakit

virus (Moss et al., 2006).

Udang vaname pertama kali diimpor ke

Indonesia pada tahun 2000 untuk mengganti

udang windu (P. monodon) yang terserang

WSSV (DKP, 2007). Pada akhir 2007, udang

ini telah dibudidayakan di lebih dari 17

provinsi di Indonesia (Taukhid dan Nur’aini,

2008). Masalah utama yang dihadapi dalam

pengembangan udang vaname adalah

penyakit terutama yang disebabkan oleh

virus. WSSV dan TSV merupakan penyakit

yang paling banyak mengakibatkan kerugian

pada industri budidaya udang vaname di

Amerika maupun Asia, termasuk di

Indonesia (Lightner, 2003). Sementara kedua

virus ini belum teratasi, kini muncul

infectious myonecrosis virus (IMNV)

sebagai penyakit baru. IMNV pertama kali

ditemukan pada tahun 2004 di Brazil, dan

pada tahun 2006 virus ini telah terdeteksi di

Indonesia (Taukhid dan Nur’aini, 2008). Saat

ini, IMNV telah menginfeksi budidaya udang

vaname di Jawa Timur, Bali, Nusa Tenggara

dan Sumatera. IMNV menyerang udang

terutama pada juvenil dan udang muda

dengan host utama adalah udang vaname.

Penyakit ini berkembang secara perlahan-

lahan dengan mortalitas kumulatif mencapai

40-70% (Lightner, 2009). Dalam mana-

jemen kesehatan budidaya udang, strategi

pencegahan penyakit dapat dilakukan melalui

berbagai cara seperti penggunaan bahan-

bahan kimia dan antibiotik, vaksinasi, bakteri

probiotik, SPF (specific pathogen free) dan

SPR (specific pathogen resistance), sistim

produksi biosekuriti, dan imunostimulan.

Penggunaan antibiotik memiliki dampak

negatif yaitu akumulasi residu dalam jaringan

ikan dan munculnya drug-resistance

pathogen. Vaksinasi meskipun sangat efektif

namun membutuhkan waktu, tenaga dan

biaya yang mahal serta proteksi yang

dihasilkan bersifat spesifik (Cook et al.,

2003). Probiotik berguna dalam mengontrol

infeksi mikroba melalui kompetisi dengan

mikroorganisme berbahaya/patogen, pro-

duksi bahan-bahan penghambat atau melalui

stimulasi sistim imune udang yang dibudi-

dayakan (Bachere, 2003). Udang SPR hanya

resisten terhadap patogen tertentu dan dengan

adanya mutasi genetik, udang SPR yang

awalnya resisten menjadi suseptibilitas

terhadap patogen yang baru. Resistensi

udang terhadap patogen juga berbeda-beda

berdasarkan siklus hidup udang. Meskipun

strategi biosekuriti seperti pengurangan

pergantian air, penyaringan, pengeringan

kolam, screening (penapisan) postlarva untuk

membatasi masuknya patogen dalam

lingkungan budidaya, dan bahkan dikom-

binasikan dengan udang SPR secara nyata

meningkatkan produksi, namun penyakit

terus saja terjadi dalam usaha budidaya

(Moss et al., 2006). Penggunaan nutrisi yang

seimbang kini sedang diteliti untuk me-

ningkatkan respons terhadap stres dan infeksi

patogen misalnya suplementasi UFA, sterol

dan vitamin dalam pakan. Pendekatan lain

adalah penggunaan imunostimulan dalam

mencegah penyakit infeksius.

Sumber imunostimulan bagi akuakultur

dapat diproduksi secara kimia atau biologi.

Bahan-bahan imunostimulator tersebut dapat

dikelompokkan berdasarkan fungsi maupun

sumbernya dan terdiri atas beragam

kelompok yakni berupa bakteri dan produk

bakteri, yeast, kompleks karbohidrat, faktor

nutrisi, ekstrak hewan, ekstrak tumbuhan,

dan obat-obatan sintetik (Sakai 1999; Sealey

dan Gatlin III 2001; Cook et al., 2003).

Penelitian ini menggunakan nukleotida

sebagai imunostimulan dalam mengontrol

penyakit pada budidaya udang vaname.

Nukleotida memiliki fungsi penting dalam

fisiologi dan biokimia seperti penandaan

(encoding) dan penerusan informasi genetik,

memediasi energi metabolisme dan cell

signalling maupun sebagai koensim,

allosteric effectors, dan cellular agonist

(Galtin III dan Li, 2007).

Nukleotida merupakan nutrien semi

esensial yang mulai mendapat perhatian

serius untuk dikembangkan penggunaannya

sebagai imunostimulan dalam budidaya ikan

dan krustasea dalam beberapa tahun terakhir

ini. Publikasi ilmiah tentang penggunaan

nukleotida pada ikan memperlihatkan bahwa

bahan ini dapat meningkatkan respons imun

Henky Manoppo et al. / Jurnal Akuakultur Indonesia 10 (1), 1–7 (2011)

dan resistensi ikan terhadap sejumlah

patogen secara simultan (Burrels et al.,

2001). Selain itu, pemberian nukleotida juga

dapat meningkatkan pertumbuhan serta

meningkatkan toleransi terhadap stress.

Pada udang, laporan-laporan penelitian

tentang penggunaan nukleotida masih belum

tersedia atau masih sangat terbatas.

Penelitian ini bertujuan untuk mengevaluasi

respons imun non-spesifik, resistensi, dan

performa pertumbuhan udang vaname yang

diberi pakan nukleotida.

BAHAN DAN METODE

Hewan uji

Juvenil udang vaname diperoleh dari

fasilitas pembesaraan udang di Bakauheni

Lampung Selatan. Udang yang diambil

dimasukkan dalam kotak styrofoam yang

dilengkapi dengan aerator baterai, kemudian

diangkut melalui jalan darat ke Laboratorium

Kesehatan Ikan Institut Pertanian Bogor.

Bahan uji

Bahan uji adalah nukleotida murni

(Sigma-Aldrich) yang terdiri atas uridine-5’-

monophosphate disodium salt, cytidine-5’-

monophosphate disodium salt, guanosine-5’-

monophosphate disodium salt, adenosine-5’-

monophosphate sodium salt, dan inosine-5’-

monophosphate disodium salt.

Persiapan pakan

Kelima jenis nukleotida dalam jumlah

yang sama dicampur terlebih dahulu secara

homogen kemudian ditimbang sesuai dosis

yang dibutuhkan yakni 400 mg/kg.

Nukleotida dicampurkan ke dalam pakan

standar dengan cara melarutkannya terlebih

dahulu dalam sedikit air, kemudian dikering-

anginkan dalam temperatur ruang. Setelah

kering, pakan dilapisi dengan albumin (putih

telur) dan dikering-anginkan kembali. Pelet

yang sudah kering selanjutnya dimasukkan

dalam kantong plastik, disimpan dalam

lemari pendingin dan siap untuk digunakan.

Prosedur penelitian dan pengambilan data

Juvenil udang vaname dipelihara selama 2

minggu dalam bak fibreglass (kapasitas 1000

L) untuk proses aklimatisasi. Selama proses

aklimatisasi, udang diberi pakan standar

dengan tingkat pemberian 3% bobot badan/

hari dan diberikan pukul 09.00, 13.00, dan

17.00 setiap hari. Kualitas air dipertahankan

stabil dan penggantian air dilakukan setiap 3-

4 hari sekali tergantung pada kondisi air yang

ada.

Udang (berat rata-rata 5,39±0,56 g)

selanjutnya dipindahkan ke dalam 6 buah

akuarium kaca (60x30x30cm) yang dileng-

kapi aerator dengan airlift system, serta

menggunakan resirkulasi air. Keenam

akuarium tersebut dibagi atas dua kelompok

masing-masing dengan 3 ulangan. Setiap unit

akuarium berisi 50 L air dengan 15 ekor

udang. Udang dalam kelompok pertama

diberi pakan nukleotida sedangkan udang

dalam kelompok kedua diberi pakan standar

tanpa suplementasi nukleotida. Pemberian

pakan dilakukan selama 4 minggu.

Selama masa percobaan, parameter

kualitas air dimonitor setiap hari untuk

menjamin agar parameter lingkungan tetap

berada dalam kondisi stabil. Kotoran dan

sisa pakan yang terakumulasi dalam

akuarium dikeluarkan melalui penyiponan.

Penggantian air juga dilakukan setiap 3-4

hari sekali tergantung pada kondisi air yang

ada.

Sampel haemolymph untuk pengukuran

parameter imun diambil dari 3 ekor udang

per unit akuarium dan dikerjakan pada akhir

periode pemberian pakan perlakuan (minggu

ke-4). Pengambilan sampel haemolymph

dikerjakan berdasarkan prosedur yang

dikemukakan oleh (Liu dan Chen, 2004).

Secara singkat, sekitar 0,1 ml haemolymph

diambil dari ventral sinus pada pangkal ruas

tubuh pertama dengan menggunakan alat

suntik 1 ml setelah sebelumnya dimasukkan

0,9 ml antikoagulan (30 mM trisodium sitrat,

0,34 M natrium klorida, 10 mM EDTA, pH

7,55, osmolaritas 780 mOsm/kg).

Parameter imun

Parameter imunitas udang yang diukur

terdiri atas total haemocyte count (THC), dan

Aktivitas phenoloxidase (PO). Prosedur

penghitungan parameter imun adalah sebagai

berikut:

Penghitungan THC

Sebanyak 50 µl campuran haemolymph-

antikoagulan dimasukkan dalam neutral

Henky Manoppo et al. / Jurnal Akuakultur Indonesia 10 (1), 1–7 (2011)

buffered formalin (10%) selama 30 menit.

Selanjutnya, THC dihitung dengan meng-

gunakan haemacytometer di bawah

mikroskop cahaya dengan pembesaran 40x.

Aktivitas PO

Aktivitas PO haemocyte diukur

berdasarkan formasi dopachrome yang di-

hasilkan oleh L-DOPA. Pengukuran aktivitas

PO dikerjakan berdasarkan prosedur yang

dikemukakan oleh Liu dan Chen, 2004.

Pertama-tama, 1 ml campuran hemolymph-

anticoagulan disentrifuse pada 700 g selama

20 menit pada 4oC. Supernatan dikeluarkan

dan pelet disuspensikan kembali secara

perlahan-lahan ke dalam larutan cacodylate-

citrate buffer (0.01 M sodium cacodylate,

0,45 M sodium chloride, 0.10 M trisodium

citrate, pH 7) dan disentrifuse kembali. Pelet

kemudian diambil dan disuspensikan dalam

200 µl cacodylate buffer (0,01 M sodium

cacodylate, 0,45 M sodium chloride, 0,01 M

calcium chloride, 0,26 M magnesium

chloride, pH 7).

Aliquot sebanyak 100 µl diinkubasi

dengan 50 µl trypsin (1 mg.ml-1 cacodylate

buffer) sebagai aktivator selama 10 menit

pada temperatur 25-26oC. Selanjutnya

tambahkan 50 µl L-DOPA (3 mg.ml-1

cacodylate buffer), setelah 5 menit,

ditambahkan 800 µl cacodylate buffer.

Optical density (OD) 490 nm diukur dengan

menggunakan Spektrofotometer (Hitachi U,

2000).

Larutan standar mengandung 100 µl

suspensi haemocyte, 50 µl cacodylate buffer

(pengganti trypsin), dan 50 µl L-DOPA

digunakan untuk mengukur background

aktivitas PO pada semua larutan uji.

Densitas optikal (OD) dari aktivitas PO pada

semua kondisi uji dinyatakan sebagai formasi

dopachrome dalam 50 µl haemolymph.

Resistensi

Setelah 4 minggu pemberian pakan

perlakuan, udang (8 ekor/akuarium) diuji-

tantang dengan bakteri Vibrio harveyi.

Sebelum dilakukan uji tantang, aerator

dimatikan terlebih dahulu selama kurang

lebih 30 menit, kemudian udang diuji tantang

melalui injeksi intramuskular 0,1 mL larutan

bakteri V. harveyi 1x106 cfu/mL pada ruas

tubuh ke tiga. Selanjutnya udang dimasukkan

kembali ke dalam akuarium. Selama periode

uji tantang, udang diberi pakan standar.

Kualitas air dimonitor agar berada dalam

kondisi stabil dan penggantian air dilakukan

setiap 3-4 hari sekali tergantung pada kondisi

air. Udang mati dikeluarkan setiap hari guna

mengkonfirmasi bahwa penyebab kematian

adalah V. harveyi. Pengamatan terhadap

mortalitas dilakukan setiap hari selama 14

hari setelah uji-tantang. Resistensi udang

diukur berdasarkan tingkat kelangsungan

hidup (SR) yang dicapai sampai pada akhir

periode pengamatan, SR dihitung dengan

formula (Effendie, 2002), SR (%)= Nt/No

x100, Nt=jumlah udang hidup pada waktu t

(ekor), No=jumlah udang hidup waktu tebar

(ekor).

Pertumbuhan

Pertumbuhan udang diukur setiap dua

minggu sekali yakni pada hari ke 14 dan 28.

Pertumbuhan dinyatakan sebagai selisih

antara berat udang yang diukur pada akhir

percobaan dengan berat udang pada awal

percobaan (Effendie, 2002): G= Wt–Wo,

G=pertumbuhan, Wt=berat udang pada

waktu t (g), Wo=berat udang pada awal

percobaan (g).

Analisis data

Evaluasi perbedaan responss imunitas

udang (THC, aktivitas PO), resistensi dan

pertumbuhan akibat adanya perlakuan

dilakukan melalui analisis ragam (Anova)

HASIL DAN PEMBAHASAN

Total haemocyte count (THC)

Suplementasi nukleotida dalam pakan

dapat meningkatkan jumlah haemocyte

udang. Hasil analisis memperlihatkan bahwa

THC udang yang diberi pakan nukleotida

berbeda sangat nyata (P<0,01) jika

dibandingkan dengan udang yang hanya

diberi pakan standar (Gambar 1).

Hasil penelitian menunjukkan bahwa

apabila udang diberi pakan nukleotida 400

mg.kg-1 pakan selama 4 minggu berturut-

turut, maka THC dapat meningkat mencapai

87% lebih tinggi dibandingkan dengan udang

yang diberi pakan standar (Gambar 1). Hasil

yang sama juga ditemukan pada penelitian

Henky Manoppo et al. / Jurnal Akuakultur Indonesia 10 (1), 1–7 (2011)

yang telah dilakukan sebelumnya yaituTHC

meningkat sekitar 76% lebih tinggi dari

udang kontrol (Manoppo et al., 2009).

Peningkatan jumlah THC disebabkan karena

nukleotida merupakan nutrien semi esensial

yang dibutuhkan untuk pertumbuhan dan

perbanyakan sel (Barnes, 2006). Sajeevan et

al. (2006) juga menyatakan bahwa nuk-

leotida yang ditambahkan dalam pakan

udang dapat mengoptimalkan fungsi

pembelahan sel termasuk sel-sel imun.

Aktivitas phenoloxidase (PO)

Pemberian secara oral nukleotida juga

meningkatkan aktivitas PO udang vaname.

Hasil analisis memperlihatkan bahwa

aktivitas PO udang yang diberi pakan

nukleotida selama 4 minggu berbeda nyata

(P<0,02) jika dibandingkan dengan udang

yang diberi pakan standar (Gambar 2).

Sampai saat ini, belum ada laporan yang

tersedia tentang pengaruh nukleotida pada

resistensi udang terhadap infeksi patogen.

Pada ikan (Li et al., 2004) melaporkan bahwa

produksi oxidative radical neutrofil darah

striped bass hibrida meningkat setelah diberi

pakan nukleotida selama 6-7 minggu dan

diuji tantang dengan Streptococcus iniae,

dan kelangsugan hidup ikan yang diberi

pakan nukleotida (80%) lebih tinggi

dibandingkan dengan ikan yang diberi pakan

tanpa suplemen nukleotida (60%).

Pada rainbow trout berukuran 53-55 g,

mortalitas ikan yang diberi pakan nukleotida

(Optimun, 2 g/kg pakan) selama 2 minggu

dan diuji tantang ISAV (infectious salmon

anaemia virus) sebesar 35,7% sedangkan

ikan yang diberi pakan tanpa nukleotida

memiliki mortalitas 48% (Burrels et al.,

2001). Sakai et al. (2001) juga melaporkan

pemberian nukleotida yang diisolasi dari

yeast RNA 15 mg/ikan selama 3 hari pada

Cyprinus carpio 100 g meningkatkan

resistensi terhadap infeksi Aeromonas

hydrophila. Aktivitas fagositosis, respiratory

burst, serum complement dan aktivitas

lisozyme ikan yang diberi pakan nukleotida

meningkat. Burgents et al. (2004) juga

melaporkan peningkatan resistensi L.

vannamei terhadap infeksi buatan Vibrio jika

udang diberi pakan mengandung

Saccharomyces cerevisiae.

Gambar 1. THC (total haemocyte count) rata-rata (x

107 sel/mL) Litopenaeus vannamei setelah diberi

pakan nukleotida selama 4 minggu.

Gambar 2. Aktivitas PO (phenoloxidase) Lito-

penaeus vannamei setelah diberi pakan nukleotida

selama 4 minggu.

Gambar 3. Kelangsungan hidup Litopenaeus

vannamei setelah diberi pakan nukleotida selama 4

minggu dan diuji tantang dengan bakteri Vibrio

harveyi 0,1x106 cfu.udang-1.

Pertumbuhan

Pertumbuhan udang diukur setiap 2

minggu sekali. Hasil penelitian menunjukkan

bahwa administrasi pakan nukleotida selama

Kelangsungan hidup (%)

Aktivitas PO

Resistensi (%)

THC (x107sel/mL)

Henky Manoppo et al. / Jurnal Akuakultur Indonesia 10 (1), 1–7 (2011)

Tabel 1. Pertumbuhan L. vannamei setelah diberi pakan nukleotida selama 4 minggu.

Perlakuan

Berat Awal (g)

Berat Akhir (g)

Perolehan Berat (g)

14 hari

28 hari

14 hari

28 hari

Pakan Standar

5,39±0,56

7,37±0,36

8,25±0,71

1,98±0,36

2,86±0,71

Nukleotida

5,39±0,56

7,71±0,81

10,12±0,57

2,32±0,81

4,73±0,57

14 hari tidak memberikan pengaruh terhadap

laju pertumbuhan udang (P>0,05). Setelah

28 hari pemberian, pertumbuhan udang yang

diberi nukleotida berbeda sangat nyata jika

dibandingkan dengan pertumbuhan udang

yang hanya diberi pakan standar (P<0,0)

(Gambar 4).

Pertumbuhan udang yang diberi nuk-

leotida mencapai rata-rata 4,73 g atau

65,38% lebih tinggi dari pertumbuhan udang

yang diberi pakan standar (Tabel 1). Hasil ini

sama seperti yang teramati pada penelitian

terdahulu dimana pertumbuhan mencapai

5,05 g atau (50.74% lebih tinggi dari kontrol)

setelah 4 minggu pemberian nukleotida 400

mg/kg pakan. Peningkatkan pertumbuhan

pada udang yang diberi nukleotida me-

rupakan hasil peningkatan efisiensi dan

pengambilan pakan udang. Adenosine dan

inosine yang sudah banyak digunakan dalam

pakan ternak, ikan dan krustasea laut yang

dapat meningkatkan pengambilan pakan

udang sehingga mengurangi leaching ke

dalam air. Sebaliknya, leaching nukleotida ke

dalam air diduga juga akan meningkatkan

daya tarik pakan dan pengambilan pakan

oleh udang. Jadi peningkatan pertumbuhan

Gambar 4. Pertumbuhan udang vaname

setelah 14 hari dan 28 hari diberi pakan

nukleotida.

udang diduga terjadi sebagai hasil pe-

ningkatan efisiensi dan pengambilan pakan

udang (Li et al., 2007).

KESIMPULAN

Pemberian nukleotida 400 mg/kg melalui

pakan selama 4 minggu memberi pengaruh

positif terhadap peningkatan respons imun

non-spesifik, resistensi dan pertumbuhan

udang vaname.

DAFTAR PUSTAKA

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Essential nutrients for shrimp growth and

immunity? Centre for Marine Studies,

University of Queensland.

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effectors in marine invertebrate: potential

tools for disease control in larviculture.

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2004. Disease resistance of Pacific white

shrimp, Litopenaeus vannamei, following

dietary administration of a yeast culture

food supplement. Aquaculture 231, 1–8.

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2001. Dietary nucleotide: a novel

supplement in fish feeds. 1. Effects on

resistance to disease in salmonids.

Aquaculture 199, 159–169.

Cook, M.T., Hayball, P.J., Hutchinson, W.,

Nowak, B.F., Hayball, J.D., 2003.

Administration of a commercial immune-

stimulan preparation, EcoActiva as a feed

supplement enhances macrophage respira-

tory burst and the growth rate of snaper

(Pagurus auratus, Sparidae (Bloch and

Schneider) in winter. Fish and Shellfish

Immunology 14, 333–345.

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Galtin III, D.M., Li, P., 2007. Nucleotide.

Departement of Wildlife and Faculty of

Nutrition, Texas A&M University System,

College Station USA.

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immune responsse of white shrimp

Litopenaeus vannamei following Vibrio

alginolyticus injection. Fish and Shellfish

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Li, P., Lawrence, A.I., Castille, F.L., Galtin

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supplement for Pacific white shrimp Lito-

penaeus vannamei (Boone). Aquaculture

Research 38, 887–890.

Li, P., Galtin III, D.M., 2006. Nucleotide

nutrition in fish: Current knowledge and

future application. Aquaculture 251, 141–

152.

Li, P., Lewis, D.H., Galtin III, D.M., 2004.

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influence immune responsses and

resistance of hybrid striped bass (Morone

chrysops x M. saxatilis) to Streptococcus

iniae infection. Fish and Shellfish

Immunology 16, 561–569.

Lightner, D.V., 2003. Exclusion of specific

pathogen for disease prevention in a

penaeid shrimp biosecurity program.

World Aquaculture Society pp. 81–116.

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ammonia on the immune responsse of

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its susceptibility to Vibrio alginolyticus.

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334.

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G., Valenzuela, M., Pascual, C., Sanches,

A., Rosas, C., 2003. Physiological,

nutritional, and immnunological role of

dietary β-glucan and ascorcic acid 2-

monophosphate in Litopenaeus vannamei

juveniles. Aquaculture 224, 223–243.

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Fatuchri, M., Harris, E., 2009. Nukleotida

meningkatkan respons imun dan performa

pertumbuhan udang vaname, Litopenaeus

vannamei. Aquacultura Indonesiana 10,

85–92.

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CA., 2006. Disease prevention strategies

for penaeid shrimp culture. The Oceanic

Institute, Hawaii USA.

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2006. Immunostimulatory effect of a

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Fenneropenaeus indicus. Aquaculture

257, 150–155.

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Ogawa, H., Tabata, M., 2001.

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isolated from yeast RNA on carp,

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Disease 24, 433–438.

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Effects of dietary nucleotides supplementation on growth, total haemocyte count, lysozyme activity and survival upon challenge with Vibrio harveyi in pacific white shrimp, Litopenaeus vannamei

Article

Full-text available

Nov 2021

The use of soybean meal (SBM) as replacement for fish meal (FM) in diets for Pacific white shrimp (PWS), Litopenaeus vannamei, involves a negative impact on the health of PWS. Dietary nucleotides modulate the immune response; therefore, they might be able to counteract this effect by enhancing PWS immunity. Based on this hypothesis, this study was aimed at evaluating the effects of nucleotide supplementation in PWS receiving diets in which FM had been partially replaced by SBM. A 70-day feeding trial was conducted to evaluate the effects of dietary nucleotides on PWS growth performance, protein levels and retention rate, total hemocyte count (THC) and lysozyme activity. Ten experimental diets were formulated. The control diet included 10% FM and 43% SBM. Another diet was formulated by reducing FM to 5% and by increasing SBM up to 50%. The rest of diets included 0.05 or 0.1% nucleotide supplementation with varying degrees of FM replacement by SBM. A total of 900 PWS post larvae with an average initial body weight of 4.24 ± 0.03 g were randomly assigned to ten study groups, with six replicates per group and 15 PWS per aquaria tank. After the performance trial, disease resistance was evaluated in a 7-day challenge test with Vibrio harveyi at the consistent concentration of 10⁵ CFU mL⁻¹. Nucleotide supplementation led to significantly higher THC and lysozyme activity (P < 0.05) and a significantly increased PWS survival in the V. harveyi challenge test (P < 0.05). No significant differences were found between groups regarding growth parameters or protein analyses (P > 0.05). In conclusion, the present study shows a positive impact of nucleotide supplementation on immune response and disease resistance against V. harveyi. Nucleotides could therefore be used as functional dietary ingredients, especially in PWS which receive diets with FM replacement by plant-protein sources.

Active Compounds on Squid (Loligo sp.) Ink Extract Powder as Immunostimulant Candidate to Against Shrimp Disease

Article

Full-text available

Dec 2019

The business of developing shrimp farming cannot be separated from the existence of disease. In shrimp farming health management, disease prevention strategies can using various methods, one of which is immunostimulant. One alternative immunostimulant source that can be used to improve the body defense system in shrimp is squid (Loligo sp.) ink extract powder who has antibacterial ability. This study aims to analyze the presence of active compound in squid ink extract powder can be used as an immunostimulant candidate to against shrimp disease. The method used in this research is descriptive explorative and experimental method. This research was conducted with several stages of squid ink extraction until it becomes powder. Identification of squid ink extract powder is using FTIR and LC-MS test. The results showed that the squid ink extract powder contained alkaloid and carboxylic acid from the FTIR test results. Based on LC-MS test results, it was found that squid ink extract powder contained betaine, cinnamic acid, and choline compounds with large amounts of content. Betaine, cinnamic acid, and choline has several biological activity as antibacterial, antioxidant, antiviral, antifungal, etc. so that it can be used as an immunostimulant against shrimp disease.

Effects of dietary nucleotides on growth performance, immune response, intestinal morphology and disease resistance of juvenile largemouth bass, Micropterus salmoides

Article

Full-text available

May 2022
J FISH BIOL

An 8 week feeding trial was carried out to investigate the effects of dietary nucleotides on growth performance, intestinal morphology, immune response and disease resistance of juvenile largemouth bass, Micropterus salmoides. Five grades of dietary nucleotide levels were designed as 0, 0.2, 0.4, 0.8 and 1.2 g kg⁻¹, respectively. Each group had 3 replicates, with 40 fish in each replicate. After the feeding experiment, 15 fish from each tank were infected with Aeromonas hydrophila for 14 days. The results indicated that fish fed the diets containing 0.4, 0.8 and 1.2 g kg⁻¹ nucleotides had higher growth performance and feed utilization than those fed the control diet. Nonetheless, there were no significant differences in survival between all the groups, although fish fed the diets with all‐level nucleotides obtained higher survival than those fed the control diet. Dietary nucleotides significantly affected the superoxide dismutase, acid phosphatase and catalase activities in serum but not the malondialdehyde content. Fish fed the 0.4 g kg⁻¹ nucleotide diets had the highest fold height, enterocyte height and muscular layer thickness significantly. The average mortality of largemouth bass infected with A. hydrophila was significantly influenced by dietary nucleotides. The mortality was significantly higher in the control group (91.11%) and 0.02% nucleotide group (73.11%) followed by the other groups and lowest in the 0.8 g kg⁻¹ nucleotide group. In summary, dietary 0.4–0.8 g kg⁻¹ nucleotides promoted growth performance, enhanced immunity and improved intestinal morphology and disease resistance of largemouth bass.

Dietary nucleotide supplementation enhances the growth, immune response, and disease resistance of the juvenile giant freshwater prawn, Macrobrachium rosenbergii

Article

Full-text available

Aug 2022
AQUACULT INT

A 56-day feeding trial was conducted to study the effects of dietary nucleotides on growth performance, non-specific immunity, and disease resistance of the juvenile giant freshwater prawn, Macrobrachium rosenbergii (initial weight, 0.095 ± 0.002 g). Five nucleotide levels were set, 0 g/kg, 0.2 g/kg, 0.4 g/kg, 0.8 g/kg, and 1.2 g/kg, respectively. Each group had 3 replicates with 300 prawns per replicate. After the feeding trial, 40 prawns from each pond were challenged with Vibrio anguillarum for 7 days. The results showed that the final body weight (FBW) and weight gain (WG) of 0.4 g/kg nucleotide group were significantly higher than those of the control group. The survival (SR) of 0.2 g/kg nucleotide group was significantly higher than that of the control group. The feed conversion ratio (FCR) of 0.4 g/kg nucleotide group was significantly lower than that of the control group. However, there was no significant difference in specific growth rate (SGR) among the experimental groups and control group. The dry matter, crude protein, and crude fat of prawns fed diet with nucleotides had no obvious trend, and there was no significant difference compared with the control group. The activities of glutamic oxaloacetic transaminase (GOT), superoxide dismutase (SOD), phenoloxidase (PO), acid phosphatase (ACP), and lysozyme (LZM) in serum of prawns were significantly influenced by the nucleotides in diet. The phagocytosis rate and phagocytic index of hemocyte were significantly higher in the nucleotide group. Finally, the average mortality of prawns infected with Vibrio anguillarum was also significantly affected by nucleotides, and the relative percent survival (RPS) was more than 50% when the nucleotide level was 0.4–1.2 g/kg. In summary, dietary with 0.2–0.4 g/kg nucleotides promoted the growth performance, enhanced the immunity, and improved the disease resistance of M. rosenbergii.

Nutritional profiling of microfungi and its effects on the growth performance, bacterial communities, and survival with Vibrio harveyi on white shrimp post-larvae (Litopenaeus vannamei)

Article

Jul 2021
AQUACULTURE

We evaluated seven microfungi isolated from rivers or streams in Indonesia for a nutritional profile including protein, lipid, amino acids, fatty acids, and beta-glucan, i.e., Trichoderma harzianum, Macrophoma theicola, Tri- choderma lentiforme, Trichoderma hamatum, Mucor circinelloides, Lasiodiplodia theobromae, and Fusarium oxy- sporum. All isolates had relatively high nutritional content, where the proteins ranging from 31.56–45.58%, lipids 2.22–6.54%, amino acids 11.023–18.881 g/100 g, fatty acids 1.094–5.253%, and beta-glucan 0.170–0.280 g/dry weight. The most amino acids were glutamic acid, lysine, and leucine, while fatty acids were oleic acid (C 18:1ω9C), linoleic acid (C18:2ω6), and linolenic acid (C 18:3ω3 and ω6). Moreover, we have conducted a 14-day feeding trial on white shrimp post-larvae (PL-10) with two isolates of microfungi and their combination, including M. circinelloides and T. harzianum. Finally, the results indicated that T. harzianum fed with 0.5% and 1.5% could improve white shrimp post-larvae survival and growth performance. It was found that two dominant bacteria were Gordonia polyisoprenivorans and Croceibacter atlanticus. The cumulative mortality rate was not significant from the challenge trial with Vibrio harveyi (p > 0.05). However, further in-depth investi- gation needs to be conducted, and some microfungi contain anti-nutrients that can affect growth.

Assessing the efficiency of a recirculating fluidized bed biofilter in white leg shrimp (Litopenaeus vannamei) broodstock aquaculture

Article

Full-text available

Jan 2020

This paper shows the results of the efficiency of a circulating fluidized bed biofiltration system using polyethelene as a filtering material. Changes in water quality parameters and the growth rates of shrimp (broodstock and commercial production) were used to assess the efficiency of water treatment system. The concentration of NH4+ in waste water from the broodstock tanks decreased 61% after three months of treatment. The final concentration of NH4+ after treatment was 0.07 mg/ml which meet the Vietnamese regulations for discard of aquaculture waste into coastal waters. Biological oxygen demand decreased 48%. The corresponding figures for treating waste water from commercial shrimp tanks were decreases of NH4+ by up to 61% with outlet concentration 0.07 mg/ml accepted to discard according to Vietnamese Standard and biological oxygen demand by 55%. Outlet water providing from broodstock and commercial shrimp tanks were treated with COD concentration decreased from 3.12 down to 2.62 mg/ml and 4.62 down to 2.68 mg/ml, or 16% and 41.9% of COD removal, respectively.

Dietary guanosine‐monophosphate improves growth performance, feed utilization and intestinal morphology of whiteleg shrimp (Litopenaeus vannamei) maintained on soybean meal‐based diets

Article

Nov 2020

The present study evaluated the effects of dietary supplementation of individual or a combination of nucleotides in soya bean meal‐based diet on growth, survival, apparent digestibility and intestinal condition of whiteleg shrimp, Litopenaeus vannamei. The experimental protocol comprised a control set, and four test diets that were separately supplemented with 0.1% inosine monophosphate (IMP, D + I), 0.1% guanosine monophosphate (GMP, D + G), 0.1% mixture of IMP and GMP (D + IG) and 0.1% mixture of IMP, GMP, cytidine monophosphate (CMP) and uridine monophosphate (UMP) (D + IGCU). Triplicate groups of the shrimp (initial body weight = 0.99 ± 0.01 g) were fed four times daily for 8 weeks. Results demonstrated that D + G significantly enhanced body weight gain and specific growth rate in the shrimp compared with control and other dietary treatments with the various combinations of nucleotides (p < 0.05). However, the survival was not significantly affected by these treatments (p > 0.05). Shrimp fed D + G showed significantly higher feed intake and apparent net protein utilization compared with D + IGCU and, control and D + IG (p < 0.05) respectively. Nucleotide supplementation did not significantly improve the crude protein and lipid digestibility coefficients. This was evident when the data on these parameters were compared with those of the control. Shrimp fed D + G showed significantly improved intestinal villus height and body crude protein content (p < 0.05). In conclusion, GMP supplementation in soya bean meal‐based diet enhanced the growth, feed intake, protein utilization and villus height of L. vannamei.

Recent advances of nucleotide nutrition research in aquaculture: a review

Article

Jul 2019

Although nucleotides (NT) have long been implicated as feed attractants in both vertebrate and invertebrate species, research into potential growth and health benefits of dietary NT in aquaculture species has just been implemented in early 2000s. To date, research pertaining to NT supplementation in aquafeed as functional nutrients has shown rather consistent and encouraging, beneficial results in fish and shrimp health management. Dietary NT serve a marked role to enhance growth in early stages of development, improve reproductive performances of brood fish, enhance larval quality, increase stress and disease resistance, modulate immune functions and improve intestinal morphology and gut microbiota of fish and shrimp. NT have also been used as functional supplement in alternative protein based diet, where NT was found as an effective supplement to increase the efficiency of utilizing alternative proteins. However, still there are numerous gaps in existing knowledge about exogenous NT administration to fish and shrimp including various aspects of digestion, absorption, metabolism, administration duration and age/size‐related responses. Adoption of different molecular technologies, like proteome modification and study of several gene expression pattern related to growth, immunity, gut health and stress resistance in details are important for exploring the underlying mechanisms of NT functions in improved growth and health performances of fish and shrimp. Efforts are also needed on production technologies to reduce the NT cost. Moreover, extensive research information is also required regarding the use of NT in alternative protein and lipid based diet to develop cost‐effective, ecofriendly functional aquafeed in near future.

Effects of yeast hydrolysate supplementation in low-fish meal diets for pikeperch

Article

Jun 2023
ANIMAL

Plant proteins have been increasingly used as sustainable substitutes for fish meal (FM) in aquafeeds; however, their high inclusion level compromises fish performance. The objective of this study was to examine whether yeast hydrolysate (YH) supplementation can improve the utilisation of high soybean meal (SM) diet and ameliorate its potential deteriorating impacts in pikeperch (Sander lucioperca). A basal diet was formulated using 44% FM, and four additional diets were produced by replacing 30 or 60% of FM with SM with or without the addition of 2% YH (FM, SM30, SM60, SM30 + YH, and SM60 + YH diets). Each diet was fed to three groups of fish (35.3 ± 0.10 g, 150 fish per group) to visual satiety four times daily for 70 days. Fish growth was not impacted by FM replacement level or YH application. However, SM60 group exhibited markedly higher feed conversion ratio and lower survival rate than those fed the FM- and YH-supplemented diets (P

Antibacterial activity of the epidermal mucus of Nile tilapia (Oreochromis niloticus) against pathogens in vannamei shrimp (Litopenaeus vannamei Boone)

Article

Full-text available

Jan 2021

Physiological, nutritional, and immunological role of dietary ?? 1-3 glucan and ascorbic acid 2-monophosphate in Litopenaeus vannamei juveniles

Article

Full-text available

Jun 2003
AQUACULTURE

The present study was designed to determine the effect of dietary β 1-3 glucan (BG) and a megadose of vitamin C on the nutrition (growth and survival, blood glucose, lactate, acylglycerides, cholesterol, hemocyanin [Hc], digestive gland glycogen [DGG]) and the immunological system (blood cells, prophenoloxidase [ProPO]) in Litopenaeus vannamei juveniles. Each treatment was replicated five times. Nutritional condition and immunological response of shrimp were recorded after 40 days of feeding with specific diets and during 48 h after a salinity shock (35–0‰). A significantly greater growth rate was observed in shrimp fed with BG or vitamin C diets than in the control group. Higher blood protein, total blood cells, granular cells, and ProPO activity were recorded in shrimp fed with vitamin C as compared to the remaining treatments. That means that BG was degraded in the digestive gland by β-glucanases to produce energy, permitting the use of more proteins for growth, whereas vitamin C was used to improve animal health, enhancing general metabolism in shrimp. The salinity stress induced a rapid use of reserves, triacylglycerols (TAGs), and cholesterol were reduced in blood and probably used as a source of energy or synthesis in the digestive gland. The increase in digestive gland glycogen 24 h after the salinity shock evidenced the use of these reserves by shrimp fed with all diets. In general, after the salinity, shock blood cells increased in shrimp fed with glucans and decreased in shrimp fed with vitamin C, whereas ProPO decreased in all shrimp after the salinity shock. This could mean that after the salinity, shock shrimp fed with glucans could synthesize cells and ProPO, whereas in shrimp fed with vitamin C, blood cells were just used to respond to the stress. After the salinity shock, an increment in the ProPO/granular cell ratio was observed in shrimp fed with vitamin C, indicating that these shrimp could rapidly increase the components of their immune system. In contrast, a continuous reduction in ProPO/granular cell ratio was observed in shrimp fed with glucans, revealing that with this type of immunostimulant, shrimp drive their immunological equipment to respond continuously to the stress. These results could explain why shrimp fed with immunostimulants presents immunological fatigue in contrast to shrimp fed with additives that improve their nutritional status, like vitamin C.

Disease prevention strategies for penaeid shrimp culture

Article

Full-text available

Jan 2003

Penaeid shrimp aquaculture expanded significantly over the past two decades. However, shrimp farmers have suffered significant economic losses because of viral diseases. Researchers from the U.S. Marine Shrimp Farming Program (USMSFP) have developed novel approaches to mitigate the devastating impact of shrimp viruses, including the use of specific pathogen free (SPF) and specific pathogen resistant (SPR) shrimp, as well as the establishment of biosecure production systems that rely on pathogen exclusion. These approaches have evolved over the past decade in response to changing disease problems faced by U.S. shrimp farmers. In the late 1980's and early 1990's, U.S. farmers observed Runt Deformity Syndrome (RDS), an economically significant and frequent disease problem of cultured Pacific white shrimp, Litopenaeus vannamei. RDS is characterized by reduced growth rates and cuticular deformities and is caused by Infectious hypodermal and hematopoietic necrosis virus (IHHNV). The increasing incidence of RDS on commercial farms catalyzed USMSFP researchers to develop SPF stocks of L. vannamei that were free of IHHNV. High health offspring from these SPF stocks were made available to U.S. shrimp farmers, resulting in a significant increase in U.S. farmed shrimp production from 1992 -1994. However, in mid-1995, Taura syndrome virus (TSV) was identified in south Texas, the major shrimp farming region in the U.S., and the resulting TSV epizootic contributed to a 164% decline in Texas shrimp production from 1994 to 1995. USMSFP researchers responded by initiating a selective breeding program to develop TSV-resistant L. vannamei. The use of these high-health SPR stocks, in conjunction with on-farm biosecurity practices, resulted in incremental increases in U.S. shrimp production from 1998 to the present. Although TSV-resistant shrimp improved production and profitability for those farmers who were experiencing crop losses from TSV, breeding shrimp for resistance to a single viral pathogen, using current selective breeding strategies, may not be the most effective course of action for the long-term viability of the shrimp farming industry. USMSFP researchers are now developing biosecure shrimp production systems which rely on pathogen exclusion, and research results indicate that it is possible to produce > 5 kg of market-sized shrimp (~ 20 g) per m 2 of raceway in about 12 weeks, using < 400 L of water per kg of shrimp. With advanced biosecure technologies available, the U.S. shrimp farming industry will be able to expand into areas away from the coast with greater control against the spread of disease and without adversely affecting the environment.

Immunostimulatory effect of a marine yeast Candida sake S165 in Fenneropenaeus indicus

Article

Full-text available

Jun 2006
AQUACULTURE

The efficacy of a marine yeast Candida sake as source of immunostimulant to Indian white shrimp Fenneropenaeus indicus was estimated. Biomass of C. sake was prepared using malt extract agar and incorporated at graded levels into a standard diet to prepare yeast diets of varying biomass concentrations (1%, 10% and 20%). F. indicus were fed on these diets for a period of 28 days and challenged orally with white spot syndrome virus (WSSV) and immune parameters such as total haemocyte count, phenoloxidase and nitroblue tetrazolium reduction (NBT) were determined. Ten per cent C. sake in the diet was found to support an optimum immune response in the animals in general and their enhancement could be observed on the second and third day following challenge with the virus. The study has demonstrated that marine yeast C. sake at 10% in diet (w/w) may be used as an effective source of immunostimulants in F. indicus.

Dietary nucleotides: a novel supplement in fish feeds: 1. Effects on resistance to disease in salmonids

Article

Jul 2001
AQUACULTURE

To determine the effects of the inclusion of exogenous nucleotides in aquaculture diets on the resistance of fish to various challenge infections, trials were carried out and results presented. When added to normal fish feed formulations at a combined inclusion level of 0.03%, these additional nucleotides were shown to increase resistance to challenge infections with bacterial, viral and rickettsial diseases as well as ectoparasitic infestation. When fed for 3 weeks prior to challenge, the nucleotide-supplemented diet was superior (31% mortalities; relative percent survival: RPS=37%) to a β-glucan-containing diet (43% mortalities; RPS=12%) in reducing mortalities due to Vibrio anguillarum infection in fish fed the control diet (49% mortalities). Mortalities resulting from infections with infectious salmon anaemia (ISA) virus and Piscirickettsia salmonis were also significantly reduced (RPS=25.7% and 42.1%, respectively) as were the numbers of infesting sea lice (Lepeophtheirus salmonis) (37.8% reduction in the mean number of attached lice per fish). The mode of action of supplemental dietary nucleotides and advantages over recognised immunostimulants are discussed.

Current research status of fish Immunostimulants

Article

Mar 1999
AQUACULTURE

Masahiro Sakai

Immunostimulants are valuable for the control of fish diseases and may be useful in fish culture. The immunostimulatory effects of glucan, chitin, lactoferrin and levamisole for fish and shrimp have been reported. Nutritional factors such as Vitamins B and C, growth hormone and prolactin have also been reported to be immunostimulators. These immunostimulants mainly facilitate the function of phagocytic cells and increase their bactericidal activities. Several immunostimulants also stimulate the natural killer cells, complement, lysozyme and antibody responses of fish. The activation of these immunological functions is associated with increased protection against infectious disease. Resistance to bacterial pathogens such as Vibrio anguillarum, V. salmonicida, Aeromonas salmonicida, Yersinia rukeri and Streptococcus spp. and to parasitic infections such as white spot disease can be increased by administration of immunostimulants, but not to intracellular pathogens such as Renibacterium salmoninarum and Pasteurella piscicida. The most effective method of administration of immunostimulants to fish is by injection. Oral and immersion methods have also been reported, but the efficacy of these methods decreases with long-term administration. Overdoses of several immunostimulants induce immunosuppression in fish. The side effects of immunostimulants have not been well-studied. Growth-promoting activity has been noted in fish or shrimp treated with glucan or lactoferrin. Immunostimulants can overcome immune suppression by sex hormones. Thus, the influence of immunostimulants in mature fish should be studied. In conclusion, immunostimulants can reduce the losses caused by disease in aquaculture; however, they may not be effective against all diseases. For the effective use of immunostimulants, the timing, dosages, method of administration and the physiological condition of fish need to be taken into consideration.

Immunostimulant effect of nucleotide isolated from yeast RNA on carp, Cyprinus carpio L

Article

Jul 2008
J FISH DIS

Nucleotides from yeast RNA were evaluated for their ability to enhance the non-specific immune responses in carp, Cyprinus carpio. Oral administration of nucleotides to fish daily for 3 days resulted in enhanced responses of phagocytic and nitroblue tetrazolium (NBT) activities in kidney phagocytic cells. This activation of kidney cells was observed for at least 10 days post-treatment. The serum complement and lysozyme activities also increased in fish treated with nucleotides. Furthermore, the number of Aeromonas hydrophila in nucleotide-treated fish significantly decreased in the blood, kidney and liver after intraperitoneal injection. Thus yeast nucleotides appear to enhance non-specific immune responses in fish.

Preliminary evaluation of a purified nucleotide mixture as a dietary supplement for Pacific white shrimp Litopenaeus vannamei (Boone)

Article

May 2007
AQUAC RES

Selection of probiotic bacteria and study of their immunostimulatory effect in Penaeus vannamei

Article

Apr 2004
AQUACULTURE

The objective of this work was to obtain probiotic bacterial strains with immunostimulatory qualities for shrimp. A total of 80 strains were isolated from the hepatopancreas of healthy wild shrimp (30±5 g) collected in Manglaralto-Ecuador. The probiotic effect in vitro was evaluated using the agar diffusion technique. Three strains identified as Vibrio P62, Vibrio P63 and Bacillus P64, showed inhibitory effects against Vibrio harveyi (S2). The colonization percentage in shrimp hepatopancreas was analysed using random amplified polymorphic DNA (RAPD) profiles with three primers. The strains P62, P63, and P64 achieved colonization percentages of 83%, 60% and 58%, respectively. The competitive interaction with V. harveyi (S2) was evaluated in shrimp using RAPDs and monoclonal antibodies. The inhibition percentage against S2 reached by strains P62, P63 and P64 was 54%, 19% and 34%, respectively. Histopathology was carried out after the colonization and interaction experiments, and confirmed that the probiotic strains had no pathogenic effects on the host. The immunostimulatory effect of P62 and P64 was evaluated in vivo using several immunity tests. Vibrio alginolyticus (Ili) was used as positive control. Shrimp that did not receive any probiotics served as the negative control group. The global immunity index was significantly higher (p<0.05) in the shrimps stimulated with P64 and V. alginolyticus. For the animals stimulated with P62, the immunity index was similar to the control. Mean shrimp weights for three probiotic groups were significantly higher (p<0.05) than the control. In conclusion the isolated strain Bacillus P64 showed both probiotic and immunostimulatory features, while Vibrio P62 only showed good probiotic properties.

Anti-infectious immune effectors in marine invertebrates: Potential tools for disease control in larviculture

Article

Nov 2003
AQUACULTURE

Evelyne Bachere

The aquaculture of molluscs and crustaceans is an economical activity that is threatened by the repeated appearance of new diseases, non-infectious and infectious, which affect cultured species. Difficulties in controlling diseases in marine aquaculture come mainly from the differences in susceptibility of the animals according to their developmental stage (from larvae to adults) and from the diversity of pathogens that affect them. Among the causative agents encountered in marine aquaculture, viral and bacterial problems as well as fungal diseases dominate the larval production. Methods have been proposed for the control of disease in aquaculture, such as the use of probiotic bacteria, immunostimulants or the improvement of the larval quality via the nutrition, and zootechnical progress has been made. So far, antibiotics have been intensively used as preventive and curative measures, but such practices are now questioned because regular use of these chemical products has led to the appearance of drug-resistant bacteria and can also have harmful effects on the environment. In this context, alternative treatments as well as animal health monitoring have to be established as preventive measures. In the long term, the most effective way for sustainable aquaculture production will certainly rely upon the production of animals selected for increased resistance to disease, but until now, few papers can be found on this topic for marine invertebrates. Today, immune criteria as enhancement of non-specific defence responses are being considered.Investigation of the innate immune systems of molluscs and crustaceans may give new insights into the management and control of diseases in aquaculture. In particular, antimicrobial peptides present various application possibilities by their use as therapeutic agents, as tools for monitoring the health status of cultured animals, and by the use of encoding genes as selection markers for improving resistance to infections.

Beta glucan binding protein and its role in shrimp immune response

Article

Nov 2000
AQUACULTURE

Despite their relatively short life and assumed lesser complexity, crustaceans have mechanisms to detect foreign matter. In particular, they appear to recognize common characteristics present in bacteria and fungi, such as lipopolysaccharides (LPS) and β-glucans. Although these microbial components can directly activate defensive cellular functions such as phagocytosis, melanization, encapsulation and coagulation, plasma recognition proteins amplify these stimuli. Beta glucan binding protein (BGBP) reacts with β-glucans and the glucan–BGBP complex induces degranulation and the activation of prophenoloxidase (proPO). This protein is present in all crustaceans studied so far and is highly conserved. Together with LPS-binding agglutinin, BGBP stimulates cellular function only after its reaction with LPS or β-glucans, resembling the secondary activities of vertebrate antibodies.

Enhancement of non-specific immune response, resistance and growth of (Litopenaeus vannamei) by oral administration of nucleotide

Abstract

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