MOLECULAR DETECTION AND CLONING FOR RICKETTSIA-LIKE BACTERIA OF MILKY HAEMOLYMPH DISEASE OF SPINY LOBSTER Panulirus spp.
Abstract: Spiny lobster (Panulirus
homarus and Panulirus ornatus) are important commodities for Indonesia. The
aquaculture of lobster is susceptible for several diseases like parasite,
fungi, bacteria, and virus. Among those diseases, milky haemolymph disease
(MHD) is often seen as a symptom to mass mortality occurred at lobster farms in
Gerupuk Bay of Lombok. The purpose of this study was to determine the lobster
diseases on cage culture in Gerupuk Bay of Lombok, West Nusa Tenggara. The
study was undertaken from January to March 2015. Diseases status was determined
by application of molecular plat-form, polymerase chain reaction (PCR) with
designation of specific primer for MHD (254F/R), 254F:
5’-CGA-GGA-CCA-GAG-ATG-GAC-CTT-3’ and 254R: 5’-GCT-CAT-TGT-CAC-CGC-CAT-TGT-3’
with PCR size product of 254 bp. and for cloned the pathogen was used
TA-cloning Invitrogen for the DNA plasmid as positive control for other
analysis. Several tissue samples i.e hepatopancreas, haemolymph, part of muscle
hepatopancreas P. homarus and P. ornatus were taken from cage culture farms at
Gerupuk Bay then preserved on 90% ethanol for further analysis by PCR and then
the amplificated DNA were cloned into pCR®2.1 plasmid and transformed into
competent E. coli. The result showed that almost all lobster samples from
Gerupuk Bay were positive infected by MHD, as the results of PCR amplification
whereas the band appeared at 254bp. Also MHD plasmid has been successfully
cloned and will be used for further examination. Histopathologically in
hepatopancreas infection have seen necrosis that contain numerous of
rickettsia-like bacteria.
Keywords: lobster; diseases;
MHD; rickettsia
Author: Isti Koesharyani, Lila
Gardenia, Ni Luh Anggra Lasmika
Journal Code: jpperikanangg160063
