White-spot syndrome virus (WSSV) introduction into the Gulf of Mexico and Texas freshwater systems through imported, frozen bait-shrimp.

We analysed 20 boxes of, frozen imported bait-shrimp (China: Parapenaeopsis sp. and Metapenaeopsis sp.) and 8 boxes of native, frozen bait-shrimp (Gulf of Mexico: Litopenaeus setiferus and Farfantepenaeus duorarum) by RT-PCR or PCR for Taura syndrome virus (TSV), yellowhead virus/gill-associated virus (YHV/GAV), white-spot syndrome virus (WSSV) and infectious hypodermal and hematopoietic necrosis virus (IHHNV). All 28 boxes of shrimp were negative for TSV, YHV/GAV and IHHNV; 2 boxes of imported bait-shrimp were WSSV-positive by 3 different PCR assays. Intramuscular injection of replicate groups of SPF (specific pathogen-free) L. vannamei juveniles with 2 different tissue homogenates prepared from the 2 WSSV-positive bait boxes resulted in 100% mortality of the test shrimp within 48 to 72 h post-injection. No mortality occurred among injected negative control groups. Histological and in situ hybridization analyses of 20 moribund treatment-shrimp demonstrated severe WSSV infections in each sample. Oral exposure of SPF L. vannamei postlarvae, PL (PL 25 to 30 stage; approximately 0.02 g) to minced tissue prepared from the 2 WSSV-positive bait-lots did not induce infection, possibly because of an insufficient infectious dose and/or viral inactivation resulting from multiple freeze-thaw cycles of the bait-shrimp during PCR testing. Use of an electric drill and collection of drill-tailings (tissue from approximately 20 to 30 shrimp) from frozen blocks of shrimp was successfully employed as an alternate tissue-sampling method without thawing. Our findings indicate that imported WSSV-infected bait shrimp, originating from China, are being sold in Texas for the purpose of sport fishing and represent a potential threat to freshwater and marine crustacean fisheries, as well as to coastal US shrimp farms.