A Novel Bunyavirus Discovered in Oriental Shrimp (Penaeus chinensis)

Herein, we describe a novel bunyavirus, oriental wenrivirus 1 (OWV1), discovered in moribund oriental shrimp (Penaeus chinensis) collected from a farm in China in 2016. Like most bunyaviruses, OWV1 particles were enveloped, spherical- to ovoid-shaped, and 80–115 nm in diameter. However, its genome was found to comprise four segments of (-)ssRNA. These included an L RNA segment (6,317 nt) encoding an RNA-directed RNA polymerase (RdRp) of 2,052 aa, an M RNA segment (2,978 nt) encoding a glycoprotein precursor (GPC) of 922 aa, an S1 RNA segment (1,164 nt) encoding a nucleocapsid (N) protein of 243 aa, and an S2 RNA segment (1,382 nt) encoding a putative non-structural (NSs2) protein of 401 aa. All the four OWV1 RNA segments have complementary terminal decanucleotides (5′-ACACAAAGAC and 3′-UGUGUUUCUG) identical to the genomic RNA segments of uukuviruses and similar to those of phleboviruses and tenuiviruses in the Phenuiviridae. Phylogenetic analyses revealed that the RdRp, GPC, and N proteins of OWV1 were closely related to Wēnzhōu shrimp virus 1 (WzSV-1) and Mourilyan virus (MoV) that infect black tiger shrimp (P. monodon). Phylogenetic analyses also suggested that OWV1 could be classified into a second, yet to be established, species of the Wenrivirus genus in the Phenuiviridae. These wenriviruses also clustered with Wenling crustacean virus 7 from shrimps and bunya-like brown spot virus from white-clawed crayfish. Of note there were no homologs of the NSs2 of OWV1 and MoV/WzSV-1 in GenBank, and whether other crustacean phenuiviruses also possess a similar S2 RNA segment warrants further investigation. In addition, we established a TaqMan probe-based reverse-transcription quantitative PCR method for detection of OWV1, and it was detected as 1.17 × 102—1.90 × 107 copies/ng-RNA in gills of 23 out of 32 P. chinensis samples without an obvious gross sign. However, the discovery of OWV1 highlights the expanding genomic diversity of bunyaviruses.

Optimization of ultrasonic-assisted freezing of Penaeus chinensis by response surface methodology

Objectives Optimization of ultrasonic-assisted freezing of Penaeus chinensis by response surface methodology was studied in order to 1. , obtain frozen Penaeus chinensis of high quality and 2. , provide practical guidance for the application of ultrasonic-assisted freezing in Penaeus chinensis. Materials and Methods Three independent and major variables were selected, including initial ultrasonic temperature (°C), ultrasonic power (W) and ultrasonic time (s on/2 s off). On the basis of one-factor experiments, 17 groups of experiments were established by response surface methodology according to box-Behnken design. Using multiple regression analysis fitted the experimental data into a second-order polynomial equation, which was tested by proper statistical methods. Results The optimal ultrasonic conditions were as follows: initial ultrasonic temperature 0 °C, ultrasonic power 180 W, ultrasonic time 5 s on/2 s off. Under the optimization conditions, the time of passing through maximum ice crystal generation zone was 105.500 s, which was very close to the predictive passage time of 101.541 s. Conclusions Initial ultrasonic temperature, ultrasonic time and ultrasonic power played an important role in the process of ultrasonic-assisted freezing of Penaeus chinensis. Response surface methodology was used to optimize the three factors in ultrasonic-assisted freezing, which could greatly shorten the time of passing through the maximum ice crystal generation zone and maintain the tissue structure of Penaeus chinensis well.