Romi Novriadi1 and KB Haw2

1) Batam Mariculture Development Centre, Ministry of Marine Affairs and Fisheries, Indonesia

2) Engineering Consultant at Aquacultural Fresh and Marine Exploratory,Singapore

Immunostimulation Effects of Herbal Bio -Conditioners on Tiger Grouper (E. fuscoguttatus)

Against V. parahaemolyticus infection

Masalah Pada Budidaya Ikan Kerapu

1. Kematian akibat penyakit, disebabkan oleh : - Jamur : Ichthyophonus sp - Bakteri : Vibriosis (including: V. parahaemolyticus) infection, Pseudomonas infection, Streptococcal infection

Flexibacter infection and Bacterial Gill Diseases - Virus : Viral Nervous Necrosis (VNN), Iridovirus - Parasit : Protozoans, Monogeneans, Nematodes, Copepods and leeches

2. Degradasi kualitas lingkunganToksisitas logam berat, air limbah, pakan yang tidak dikonsumsi, ….dsb

New approach : Aplikasi pengobatan herbal

Apa keuntungan aplikasi herbal pada akuakultur?

Immune enhancing activityHelp to cure various diseases

Antimicrobiological activities

Mengandung berbagai senyawa aktif seperti: alkaloids, flavonoids, pigments, phenolics, terpenoids, steroids dan essential oils

Tujuan Studi

Melakukan analisa terhadap pengaruh aplikasi ekstrak bakau dan ketapang untuk memperkuat sistem imun dan tingkat kelulushidupan Kerapu macan (E. fuscoguttatus) melawan infeksi V. parahaemolyticus

Studi awal di BPBL Batam Studi lanjutan di Aquacultural F.A.M.E(s).

Proses Ekstraksi Bahan Herbal

Daun bakau dan ketapang

DikeringkanDihancurkan dengan mechanical grinder (tepung)

Dimasukkan dalam tabung V distiller

5 gr tepung herbal dilarutkan dalam 50 mL methanol dan aceton

Ekstraksi dilakukan selama 48 jam (100 rpm)

Larutan ekstraksi

(+) osmoregulatory salt: CaCO3, MgSO4, KCl

Evaporasi (40 -72 C)⁰ ⁰

Crude extract

Experimental design

Artemia Feeder Inlet

Air stone

Outlet

Bio-security

1-2 cm of Tiger grouper

175 L saline water (30 mg/L)

29±10 C using thermostatic heater

Exchanged ± 50%/day

Density:3 fish/L

NH4-N, NO2-N, and NO3-N were measured every two days

DO > 5 mg/L

Experimental Design (Bacterial Culture)

Bacterial Culture± 150 µl of bacteria strain aseptically inoculated in

30 ml marine broth

Incubated overnight at 250-280 C Challenge bacteria

Bacteria as feed

Autoclaved at 121 C and 15 psi for 20 minutes

150 µl was subsequently transferred and grown again in 30 ml of marine broth 6 hours before challenge

Density Measurement at

550 nm

Axenity Verification

100 µl 5 days incubation

Experimental Design

1-2 cm of Tiger

grouper

Conditioning for 6 h with 0.05 g/L

AquaHerb

Challenged with Vibrio parahaemolyticus (105

cells/ml)

Washing Medication (AquaHerb Immersion) 0.05 g/L

24 h

72h

1. Survival (%)2. Immune response 12 h

6 h

72h

72h

Control: Exposed to sea water for 6 h

Survival Percentage (%)

Fig 1: Histogram of the mean survival (%) of tiger grouper challenged with V. parahaemolyticus at 105 cells/ml. Survival was scored at 72 hours after AquaHerb immersion and the addition of V. parahaemolyticus for the challenge.

Each experiment was repeated twice and similar results were observed. Significant differences among the treatments and control at corresponding time of herbal exposure are indicated by different letter ( p< 0.05).

dc

e

b

f

a

Immune Response: Leukocytes

a

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c

cd dcd

d d

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Immune Response: Monocytes (%)

a

bab

abab

ab

bb b b

b b

Immune Response: Lymphocyte (%)

aa a

b bbbb b

abab ab

Immune Response: Neutrophile (%)

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Conclusion1. Herbal Bio Conditioners merupakan senyawa dan teknik

immunostimulan yang baik dan mampu meningkatkan kelulushidupan kerapu macan melawan infeksi Vibrio parahaemolyticus

2. Herbal Bio Conditioners mampu meningkatkan parameter imun pada ikan kerapu macan (Leukocytes, Monocytes, Lymphocyte and Neutrophile)

Further Research

Perlu dilakukan analisa sistem imun spesifik dan non-spesifik lainnya sebagai dampak aplikasi Herbal Bio Conditioners serta perlakuan aplikasi herbal untuk waktu yang lebih lama Immunosupression ????

TERIMA KASIHPrevention is Better Than Cure

Lancang kuning lancang pusaka Nampak dari Tanjung Puan Kalau Antibiotik jadi Petaka

Baiklah Herbal Dijadikan Pilihan