ABSTRACT-White spot syndrome associated baculovlrus (WSBV) is the causative agent of a dlsease which has recently caused high shrimp mortahties and severe damage to shrimp cultures. In thls study, a strain of WSBV from black tiger shrimp Penaeus monodon was used to develop a diagnostlc tool for the detection of WSBV and related agent lnfect~ons in shnmp The vlnons were punfied from P monodon Infected with LVSBV V~ral genomlc DNA was extracted from purlfled vinons by treatlng the vlnons \ n t h proteinase K dnd cetyltnmethylammonium bromlde (CTAB) followed by phenol-chloroform extraction and ethanol precipitation A qualitative assessment Ivas performed using polymerase chain reaction (PCR) analys~s on the viral DNA and primers specif~c to shrimp genomic DNA in order to mon~t o r shrimp DNA contamination In the viral genomic DNA preparations A WSBV genomlc DNA llbrary was constructed and based upon the sequence of the cloned WSBV DNA fragment, we deslgned a LVSBV-specific prlmer set for PCR to detect WSBV Infection in penaeld shrimp Samples which contained WSBV DNA yielded a n evident ampl~f~catlon product showing the expected moblllty of a 1447-bp DNA fragment whereas n u c l e~c aclds extracted from tissue samples of clln~cally healthy shnmp showed no such DNA fragment, thereby confirming the speclficity of our pnmers By PCR with thls prlmer set, ~t was demonstrated that the causative agents of white spot syndrome in different shnmp specles are closely related An effective diagnostlc tool is thus provided for screening shnmp for \.VSBV infections, and may be important In preventing the further spread of this d~s e a s e KEY WORDS: WSBV . W h~t e s p o t . PmNOBIII . Detection . Penaeid shnmp baculovirus . PCR

ABSTRACT. The causative viral agent was purified from diseased shrimp Penaeus monodon with white spot syndrome. Negatively stained preparations show that the virus is pleiomorphic. It is fusiform or rod-shaped. In negatively stained preparations, the virion measures 70 to 150 nm at its broadest point and is 250 to 380 nm long. In some virions, a tail-like projection extends from one end. The capsid is apparently composed of rings of subunits in a stacked series. The rings are aligned perpendicular to the longitudinal axis of the capsid. The genome of the virus is a double-stranded DNA molecule which produces at least 22 Hind 111 fragments. The full length of the DNA is estimated to be longer than 150 kbp. Based on the morphological characteristics and genomic structures of the virus, we confirm that white spot syndrome associated virus (MJSSV) is a member of genus NOB (Non-Occluded Baculovirus) of the subfamily Nudibaculovirinae of Baculoviridae, name the present isolate PmNOBIII, and propose the use of WSBV (Baculovirus associated with White Spot syndrome) to indicate PmNOBIII related agents.

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Short title: sul and tet(M) in marine aquaculture Highlights:1. sul1, sul2, sul3, and tet(M) were detected in Japanese mariculture environment.2. sul2 was constant, whereas others varied monthly.3. sul3 was present in non-culturable bacteria in seawater, but not in cultured bacteria. 4. Four resistance gene profiles among Japan, Taiwan, and Finland were different.