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Discovered Vibrio harveyi on Vannamei Shrimp Litopenaeus vannamei Infected White Feces

Disease in Situbondo, East Java


sumini1, Kismiyati2 dan Rahayu Kusdarwati2

Balai Perikanan Budidaya Air Payau Situbondo


1)

2)
Departemen Manajemen Kesehatan Ikan dan Budidaya Perairan, Fakultas Perikanan dan Kelautan Universitas
Airlangga, Surabaya
*e-mail: suminicantiex@gmail.com

Introduction
White Feces Disease (WFD) infected shrimp L. vannamei farm and causing significant economic
losses to shrimp aquaculture industry in China, Indonesia, Malaysia, Thailand, Vietnam and
other countries in southeast asia. WFD symptoms were characterized by a discoloration of pale
hepatopancreas and white feces presented, floating on the water pond surface, as well as the
discoloration of intestine that became white, growth retardation and increased FCR values.
Shrimp infected WFD with vibriosis symptoms found during shrimp culture was called as
Hepatopancreatic Septic Necrosis (SHPN). Vibrio harveyi was a Vibrio species that became the
most dangerous species, leading to a cause of death on shrimp during the larval and adult stadia.
V. harveyi is a main cause of luminescene vibriosis disease occurrence on shrimp culture in the
brackishwater pond. Vibrio bacteria found on the shrimp that have infected by WFD in Thailand.
Several species of Vibrio are V. fluvialis, V. vulnificus, V. parahaemolyticus, V. alginolyticus, V.
mimicus, V. damselae and V. chlorella.

Objectives
This research aimed to observed the infection of V. harveyi on vannamei shrimp culture in Situbondo, East
Java Province.

Materials and Method


L. vannamei infected WFD, binocular microscope (Olympus, Japan), incubator (Incucell MMM,
German), autoclave (Hirayama, Japan), analitical scale (Pioneer, OHAUS, USA), reaction tubes,
Petridish, object glass, heating block (Benchmark, USA), centrifuge (Thermo Scientific, USA), thermal
cycler (Veriti 96-Well Thermal Cycler Applied Biosystems, USA), electrophoresis set (Mupid 2 Plus,
Takara, Japan). Materials used in this research were vannamei shrimp (Litopenaeus vannamei)
infected WFD, biochemical test kit, BioMerieux Analytical Profile Index/API 20NE, Promega
GoTaq® Green Master Mix, Specific primer of V. harveyi, Promega Agarose, Peqlab peqGreen
DNA RNA Dye

Target :
V. harveyi

Identification :
Bacterial isolation from WFD Bacterial culture in TCBS and pure Biochemical Test Antibiotic Sensitivity Test
shrimp culture for each of colonies API 20NE System
PCR

RESULT
Fig. 1 Healthy Shrimp vs WFD shrimp

Fig. 2. PCR with V. harveyi specific primer. PCR test result on 17 bacterial isolates.
Line : 1, Negative control; 2, 100 bp marker; 3, positive control, 4-20, bacterial isolate.

Non Vibrio V. harveyi


11.76 Total Result
V. alginolyticus V. fluvialis
V. harveyi
20.0 5.88 58.82
17.65 17.65
Number of samples (%)

V. alginolyticus
15.0 5.88
11.76 V. fluvialis
11.76
10.0 11.76 V. parahaemolyticus
5.88
5.0 5.88 17.65 Non Vibrio
0.0
FECES INTESTINE HP WATER

Fig 3. Identification result of bacterial isolates from WFD Shrimp


100
100 100
80
Persentase Resistence (%)

80

60
Resistence
40
Intermediat
20

0
Oxytetracycline Enrofloxacin Erythromycin

Fig 4. Antibiotic Resistance of V. harveyi isolates

Table 1. Water Quality and and Plankton Abundance in L. vannamei WFD Ponds
Optimum
Parameter WFD Pond Water
(SNI, 2014)
pH 7.76 7.5-8.5
Total Alkalinity (ppm) 236 100-150
Salinity (ppt) 21 10-32
NO2- (ppm) 8.12 <1
NH3 (ppm) 3.62 <0.1
Total Organik Matter (ppm) 117.55 <90
Domination of plankton abundance Chloropyceae and Diatom

Disscusion

Vibriosis was caused due to the poor water quality, unstabilized environmental condition, high
stocking density and stress condition on the shrimp. The results showed that V. harveyi were found all
samples : water, hepatopancreas, intestine, and feces, while non Vibrio bacteria S. putrefaciens, was also
found on feces. This result was supported that in China, microbial intestinal communities in vannamei
shrimp with WFD, as decreasing the biota diversity and microbe abnormalities in the shrimp intestines
were identified with a DNA sequencing method with 16S rRNA gene target, resulting Vibrio (5.89%) and
Shewanella (1.17%) discovery. Major V. harveyi isolated in this research had developed their ability to
resist any antibiotics used. This result showed that these V. harveyi species were resistant to more than
one antibiotics.

Conclusion

 V. harveyi were discovered from vannamei shrimp infected WFD


 V. harveyi observed on WFD represented resistance level for more than one antibiotics.
 WFD was presented infecting vannamei shrimp during the poor water quality and planton abundance
in the pond