Temporal changes in transcriptome profile provide insights of White Spot Syndrome Virus infection in Litopenaeus vannamei.

Shrimp aquaculture is severely affected by WSSV. Despite an increasing effort to understand host/virus interaction by characterizing changes in gene expression (GE) following WSSV infection, the majority of published studies have focussed on a single time-point, providing limited insight on the development of host-pathogen interaction over the infection cycle. Using RNA-seq, we contrasted GE in gills of Litopenaeus vannamei at 1.5, 18 and 56 hours-post-infection (hpi), between WSSV-challenged and control shrimps. Time course analysis revealed 5097 differentially expressed genes: 63 DEGs were viral genes and their expression in WSSV group either peaked at 18 hpi (and decreased at 56 hpi) or increased linearly up to 56 hpi, suggesting a different role played by these genes during the course of infection. The remaining DEGs showed that WSSV altered the expression of metabolic, immune, apoptotic and cytoskeletal genes and was able to inhibit NF-κB and JAK/STAT pathways. Interestingly, GE changes were not consistent through the course of infection but were dynamic with time, suggesting the complexity of host-pathogen interaction. These data offer novel insights into the cellular functions that are affected during the course of infection and ultimately provide a valuable resource towards our understanding of the host-pathogen dynamics and its variation with time.

Comparative analysis of shrimp (Penaeus vannamei) miRNAs expression profiles during WSSV infection under experimental conditions and in pond culture.

In recent years, the importance of viral and host microRNAs (miRNAs) in mediating viral replication and control of host cellular machinery, has been realised and increasing efforts have been taken in order to understand the interactions of miRNAs from host and pathogen during infection. However, all existing studies has thus far been conducted in controlled experimental conditions and the veracity of these data for field conditions are yet to be established. In this framework, small RNA sequencing was performed to identify the miRNAs involved in shrimp (Penaeus vannamei) immune responses under two different WSSV infection conditions of natural infection and experimentally challenged conditions. The expression profiles of miRNAs of shrimp infected with WSSV under two contrasting conditions were compared and as a result, 23365 known miRNAs and 481 novel miRNAs were identified. Amongst the most abundantly expressed miRNAs, the hypoxia related miR-210 and immune pathway related miR-29b were expressed only in infected shrimps of both conditions. miR-8-5p, having a functional role in modulation of chitin biosynthesis was exclusively represented in higher numbers in the WSSV -infected shrimps under natural conditions whilst four of the miRNAs (mja-miR-6493-5p, mja-miR-6492, mmu-miR-3968, tcf-miR-9b-5p) identified from shrimps collected from pond culture targeted chitinase, an important enzyme involved in growth and moulting in shrimps, indicating an interaction between WSSV infection and moult cycle under culture conditions. Some of the miRNAs (tca-miR-87b-3p, cte-miR-277a) and miRNAs belonging to class miR-9, miR-981 that were identified only in WSSV infected shrimps under experimental conditions, are known to respond against WSSV infection in shrimps. Moreover, the miRNA target prediction revealed several immune-related gene targets such as cathepsin, c-type lectin, haemocyanin and ubiquitin protein ligase were commonly identified under both the conditions. However, the miRNAs identified from challenge experiment had wide number of gene targets as compared to the miRNAs of natural infection. The shrimp miRNA mja-miR-6489-3p, was also found to target early virus gene wsv001 of WSSV. Our study, therefore, provides the comparative analysis of miRNA expression from shrimp during WSSV infection in two different conditions.

Retropupillary fixation of iris-claw lens in visual rehabilitation of aphakic eyes.

CONTEXT: Surgical outcome of retropupillary fixation of iris claw lens. AIMS: To evaluate the various indications, intra and post-operative complications, and visual outcome of retropupillary fixation of iris claw lens in aphakic eyes. SETTINGS AND DESIGN: The study design is a retrospective study at a tertiary eye care center. METHODS: Review of medical records of 61 aphakic eyes of 61 patients, who were rehabilitated with retropupillary fixation of an iris claw lens, with a follow-up duration of at least 1 year. STATISTICAL ANALYSIS USED: Data analysis was performed using paired t-test and Chi-square test. RESULTS: Mean preoperative uncorrected visual acuity was 1.66 ± 0.3 LogMAR and postoperative acuity at 1 year was 0.53 ± 0.5 LogMAR (P = 0.00001). Preoperative distant best-corrected visual acuity was 0.30 ± 0.48 LogMAR and postoperative acuity at 1 year was 0.27 ± 0.46 LogMAR (P = 0.07). Mean preoperative astigmatism was 1.43 ± 1.94 D and postoperatively was 1.85 ± 2.16 D (P = 0.0127). Mean endothelial cell count was 2353.52 ± 614 cells/mm2 preoperatively which decreased to 2200 ± 728 cells/mm2 at 1 year follow-up (P = 0.006). There was no significant difference in central macular thickness and intraocular pressure pre and post-surgery. Complications included ovalization of pupil in 9.83%, hypotony in 1.63%, toxic anterior segment syndrome in 1.63%, cystoid macular edema in 11.47%, epiretinal membrane in 3.27%, and iris atrophy in 6.55%. CONCLUSION: Iris claw is a safe and an effective method of rehabilitating aphakic eyes.