Abstract
White Spot Syndrome Virus (WSSV) has emerged as one of the most prevalent and lethal viruses globally and infects both shrimps and crabs in the aquatic environment. This study aimed to investigate the occurrence of WSSV in different ghers of Bangladesh and the virulence of the circulating phylotypes. We collected 360 shrimp (Penaeus monodon) and 120 crab (Scylla sp.) samples from the south-east (Cox’s Bazar) and south-west (Satkhira) coastal regions of Bangladesh. The VP28 gene-specific PCR assays and sequencing revealed statistically significant (p < 0.05, Kruskal–Wallis test) differences in the prevalence of WSSV in shrimps and crabs between the study areas (Cox’s Bazar and Satkhira) and over the study periods (2017–2019). The mean Log load of WSSV varied from 8.40 (Cox’s Bazar) to 10.48 (Satkhira) per gram of tissue. The mean values for salinity, dissolved oxygen, temperature and pH were 14.71 ± 0.76 ppt, 3.7 ± 0.1 ppm, 34.11 ± 0.38 °C and 8.23 ± 0.38, respectively, in the WSSV-positive ghers. The VP28 gene-based phylogenetic analysis showed an amino-acid substitution (E→G) at the 167th position in the isolates from Cox’s Bazar (referred to as phylotype BD2) compared to the globally circulating one (BD1). Shrimp PL artificially challenged with BD1 and BD2 phylotypes with filtrates of tissue containing 0.423 × 109 copies of WSSV per mL resulted in a median LT50 value of 73 h and 75 h, respectively. The in vivo trial showed higher mean Log WSSV copies (6.47 ± 2.07 per mg tissue) in BD1-challenged shrimp PL compared to BD2 (4.75 ± 0.35 per mg tissue). Crabs infected with BD1 and BD2 showed 100% mortality within 48 h and 62 h of challenge, respectively, with mean Log WSSV copies of 12.06 ± 0.48 and 9.95 ± 0.37 per gram tissue, respectively. Moreover, shrimp antimicrobial peptides (AMPs), penaeidin and lysozyme expression were lower in the BD1-challenged group compared to BD2 challenged shrimps. These results collectively demonstrated that relative virulence properties of WSSV based on mortality rate, viral load and expression of host immune genes in artificially infected shrimp PL could be affected by single aa substitution in VP28.
Highlights
Shrimp aquaculture is one of the major earning sources in many countries including Bangladesh and plays a vital role in enlightening community advancement, food security, employment opportunities and poverty reduction [1,2]
By comparing the virulence assay between challenged doses, we found that 66 h of challenge, and 100% mortality of post larvae (PL) occurred at 102 h after challenge with both shrimp PL challenged with Dose 1 (0.423 × 109 copies of White Spot Syndrome Virus (WSSV) per mL) started to die after phylotypes
The in vivo infection assay of the shrimp PL with BD1 phylotype showed an earlier lethal time 50 (LT50) and LT100 and higher viral load compared to those challenged with BD2
Summary
Shrimp aquaculture is one of the major earning sources in many countries including Bangladesh and plays a vital role in enlightening community advancement, food security, employment opportunities and poverty reduction [1,2]. In Bangladesh, shrimp aquaculture provides livelihood to around 85 million people (mostly coastal people) and serves as the second most foreign currency-earning source, which contributes about 5% to national. Black tiger shrimp (Penaeus monodon) contributes 26% to the total aquacultural production in Bangladesh, while crabs contribute 6% [5]. The mud crab, Scylla spp., is distributed widely throughout the Indo-Pacific region [8]. Mud crab (Scylla olivacea) farming has had an increasing trend in the coastal areas of Bangladesh due to their higher disease resistance capacity and market values [9]. Mud crabs may well suppress viral replication by inducing the apoptosis of hemocytes [13]
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