White Spot Syndrome Virus (WSSV), the etiological agent of White Spot Disease (WSD) is a major impediment for shrimp aquaculture in the worldwide. A critical threshold level of WSSV load in infected shrimp is an important trait for disease manifestation and WSSV transmission in cultured shrimp and subsequently make outbreaks. The present study investigated 120 naturally infected cultured shrimp samples by SYBR Green based qPCR assay for WSD diagnosis and quantification of WSSV load. Among them, 94 samples resulted a variable count of WSSV load ranging from 2.1 × 10⁸ to 2.64 × 10¹⁴ copies/g of shrimp tissue. The severity of WSSV infection was assessed based on the established critical threshold load of WSSV in shrimp tissue. Compared to the established critical threshold value of WSSV load in shrimp tissue, our findings showed the horrifying scenario of the severity of WSSV infection in cultured shrimps of Bangladesh that was found to be above the critical limit to initiate an outbreak in the Bangladeshi shrimp aquaculture industry. The latest phylogenetic pattern was altered from the former monophyletic history among WSSVs of Bangladesh due to a variation at 500th nucleotide of VP28 coding gene. Viruses characterized from recent outbreaks in 2015 and 2017 displayed amino acid substitution at position 167 (G→E) on the surface of VP28 protein which has demonstrated the probable replacement of indigenous virus pool. Therefore, it is imperative to take initiative for the management and prevention of WSSV outbreak to sustain shrimp aquaculture in South-West region of Bangladesh.

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.

Microbial community inhabiting the intestine of the shrimp (Penaeus monodon) and their surrounding environments (e.g., water and sediment) is considered as a key contributing factor for the sustainable farming of shrimp. Indiscriminate application of antibiotics in aquaculture is a growing concern due to the emergence of antibiotic resistant bacteria (ARB), more specifically the antibiotic resistance genes (ARGs). The present study investigates the microbiome composition and 19 ARGs from four different shrimp farming systems; (i) cluster, (ii) extensive, (iii) semi-intensive, and (iv) improved extensive in the southwest coastal region of Bangladesh. In doing so, the study applied advanced 16S rRNA-based metagenomic sequencing to study the bacterial composition. Moreover, gene specific polymerase chain reaction (PCR) was employed to detect the ARGs in shrimp, water, and sediments of different farming systems. In the current study, bacteria from the phylum Proteobacteria and Firmicutes were predominant among the samples (n = 12) collected from the different farming systems followed by Actinobacteria, Bacteroidetes, and Cyanobacteria. Firmicutes was the predominant phylum in the gut of shrimp cultured in the cluster (relative abundance 53.33%) and semi-intensive (relative abundance 59.2%) culture systems. Results indicated that the bacterial community structure was significantly ( ) distinct among gut, sediment, and water samples as well as the farming systems. The shared operational taxonomic unit (OTU) in the sediment sample (16,495) was nearly double than the gut (7,931) and water (8,513) bacterial communities. The improved extensive farming system showed 1,289 (11.05%) shared OTUs among gut, sediment, and water followed by semi-intensive (6.87%), cluster (6.27%), and extensive (5.46%) farming system. Among the tested ARGs, sul1, cat, gyrA(C), tetA, tetC, tetX, ere(A), vanR, and dfrA1 were predominant in water and sediment samples. Semi-intensive farming system had the highest prevalence of ARGs (21.05%) while the lowest prevalence was found in extensive (5.26%) farming system. Overall, the study provides a comprehensive scenario of bacterial composition and growing emergence of ARGs in shrimp farming of Bangladesh. Therefore, the production strategy must focus on the alternatives of antibiotic for shaping the shrimp cultivation technique more sustainable.

Tilapia aquaculture is emerging throughout the world because of its fast growth and adaptability. However, disease is one of the impediments in emerging this sector. Boosting the immunity of tilapia by using immunostimulants can help in fighting the diseases. This study investigates how bacterial pathogen-associated molecular patterns (PAMPs), specifically lipopolysaccharide (LPS) and peptidoglycan (PGN), influence the regulation of expression of innate immune genes in Nile tilapia. In the current study, E. coli derived LPS and PGN were injected at 100 μg LPS/kg and 100 μg PGN/kg intraperitonially in treatment group and 100 μg PBS/kg in control group. Significant upregulation of TLR2, TLR3, TLR8, and TLR9 was observed in the spleen, liver, gill, and brain at various time points post-treatment (P<0.05). Additionally, increased expression of IRF3 and TGF-β was noted in multiple organs, along with elevated levels of pro-inflammatory cytokines IL-1β and IL-10, and antiviral protein viperin. These molecular responses suggest that E. coli derived LPS and PGN can effectively modulate inflammatory pathways, enhancing the immune defense mechanisms of tilapia. The findings highlight the potential of bacterial PAMPs to improve fish health, reduce mortality rates, and increase profitability in aquaculture by reducing the reliance on antibiotics.