Circulating Phylotypes of White Spot Syndrome Virus in Bangladesh and Their Virulence

Mehedi Mahmudul Hasan; M Nazmul Hoque; Firoz Ahmed; Md Inja-Mamun Haque; Munawar Sultana; M Anwar Hossain

doi:10.3390/microorganisms10010191

Circulating Phylotypes of White Spot Syndrome Virus in Bangladesh and Their Virulence

Microorganisms. 2022 Jan 16;10(1):191. doi: 10.3390/microorganisms10010191.

Authors

Mehedi Mahmudul Hasan^{1

2}, M Nazmul Hoque³, Firoz Ahmed⁴, Md Inja-Mamun Haque⁵, Munawar Sultana¹, M Anwar Hossain^{1

6}

Affiliations

¹ Department of Microbiology, University of Dhaka, Dhaka 1000, Bangladesh.
² Department of Fisheries and Marine Science, Noakhali Science and Technology University, Noakhali 3814, Bangladesh.
³ Department of Gynecology, Obstetrics and Reproductive Health, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur 1706, Bangladesh.
⁴ Department of Microbiology, Noakhali Science and Technology University, Noakhali 3814, Bangladesh.
⁵ Department of Fisheries, University of Dhaka, Dhaka 1000, Bangladesh.
⁶ Vice-Chancellor, Jashore University of Science and Technology, Jashore 7408, Bangladesh.

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.

Keywords: VP28; WSSV; phylotypes; real-time PCR; viral load.