title: White spot syndrome virus (WSSV) PCR-positive Artemia cysts yield PCR-negative nauplii that fail to transmit WSSV when fed to shrimp postlarvae.
abstract: Positive results were obtained with nested white spot syndrome virus (WSSV) diagnostic PCR performed on 5 commercial brands of dry-packed Artemia cysts using several WSSV genomic sequence-specific primers. In 2 brands, PCR and nucleotide sequence analysis found C-->T and C-->G point mutations in the pms 146 WSSV amplicon, but in all 5 brands, the nucleotide sequences that were successfully amplified by the rrl, rr2 and tk-tmk gene-specific primer sets were identical to those of Penaeus monodon WSSV. However, despite the inarguable presence of WSSV or WSSV-like template DNA, we were unable to detect WSSV by PCR in hatched nauplii derived from PCR-positive cysts or in P. monodon postlarvae fed Artemia nauplii hatched from such cysts. Most simply, these results suggested that the cysts were externally contaminated with WSSV or WSSV-like template material that was removed during hatching and washing of the nauplii. Given the small sequence variations found, it may also have been a variety of WSSV non-infectious for P. monodon or Artemia and derived from other crustaceans or arthropods in the Artemia environment. However, we could not establish this conclusively and a small possibility remained that the PCR template in these tests was derived from WSSV template present internally in the cysts and derived from infected Artemia adults. However small, this possibility must be vigorously tested, given the impact that a positive outcome could have on the shrimp industry.
authors.affiliation: Department of Zoology, National Taiwan University, Taipei, ROC.
authors.forename: Yun-Shiang
authors.initials: YS
authors.lastname: Chang
authors.forename: Chu-Fang
authors.initials: CF
authors.lastname: Lo
authors.forename: Shao-En
authors.initials: SE
authors.lastname: Peng
authors.forename: Kuan-Fu
authors.initials: KF
authors.lastname: Liu
authors.forename: Chung-Hsiung
authors.initials: CH
authors.lastname: Wang
authors.forename: Guang-Hsiung
authors.initials: GH
authors.lastname: Kou
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017756:Aquaculture; D001156:Artemia / Q000821:virology*; D001483:Base Sequence; D017931:DNA Primers; D004267:DNA Viruses / Q000235:genetics / Q000302:isolation & purification*; D004279:DNA, Viral / Q000737:chemistry; D033364:Decapoda / Q000821:virology*; D005784:Gene Amplification; D007814:Larva / Q000821:virology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012150:Polymorphism, Restriction Fragment Length
chemical_list: D017931:DNA Primers; D004279:DNA, Viral