TITLE:
Early Detection of White Spot Syndrome Virus (WSSV) in Isolated Hemocytes of Litopenaeus vannamei
AUTHORS:
Ariadne Hernández-Pérez, Rossanna Rodríguez-Canul, Edgar Torres-Irineo, Fernando Mendoza-Cano, Daniel Eduardo Coronado-Molina, Jesús Alejandro Zamora-Briseño, Jorge Hernández-López
KEYWORDS:
WSSV, Litopenaeus vannamei, Ie1 Gene, Hemocytes, In Vitro Infection, GLS Analysis
JOURNAL NAME:
CellBio,
Vol.6 No.1,
March
31,
2017
ABSTRACT: To date, White Spot
Syndrome (WSS) produced by the White Spot Syndrome Virus (WSSV) causes one of
the most severe diseases infecting penaeid shrimps worldwide. Although a vast
amount of studies has elucidated pathogenesis in live infection models, there
is still little information about the interaction of WSSV infections using in vitro models in the whiteleg shrimp Litopenaeus vannamei (L. vannamei) hemocytes. In this study, a
WSSV infection kinetics was performed using total hemocytes isolated from
healthy L. vannamei organisms and
maintained in in vitro conditions
using isotonic solution for shrimp (ISS). The infected experimental cells
received ≈ 30,000 viral copies of WSSV. The viability of the hemocytes (control
and infected group) was measured during the kinetics with trypan blue exclusion
method and cells were maintained up to 6 hpi (post-infection) with
non-significant differences of viability between both groups. WSSV replication
was assessed using RT- PCR at the RNA expression level of the early viral gene Ie1 and transcripts were detected as
early as 30 min pi. Hemocytes from WSSV group showed disrupted integrity,
degranulation and irregular shape. This study provides evidence of the capability
of WSSV to infect and replicates in L.
vannamei hemocytes using in vitro assays in short times as 30 min.