Pyocyanin as a safe aquaculture drug for the control of vibriosis in shrimp recirculating aquaculture system (RAS)

Diseases are one of the most critical limiting factors in aquaculture. Recirculating aquaculture systems (RAS) are one of the most functionally viable sustainable aquaculture production systems adopted world over. In the event of any eventuality caused by bacterial pathogens, antibiotics or other antibacterial agents cannot be applied due to the vulnerability of biological filters which form an integral part of the RAS. Because of this, newer drugs are required for the effective control of diseases in RAS which would not interfere with the activity of nitrifying bacteria used in the biological filters. The antagonistic activity of pyocyanin, a bioactive compound produced by Pseudomonas aeruginosa, against vibrios is well established. The purpose of this study was to prove the effectiveness of pyocyanin as an aquaculture drug for application in RAS by employing a pilot-scale shrimp culture under RAS. It was noted that at the concentration of 5 mg L^?1, pyocyanin could significantly bring down the population of Vibrio spp. in RAS without affecting noticeably the other natural heterotrophic bacteria. Also, pyocyanin at this concentration did not inhibit the activity of nitrifying bacterial consortia used in the SBSBR (stringed bed suspended bioreactor) of RAS. The reared shrimp (Penaeus monodon) showed 100% survival after the application of pyocyanin, besides exhibiting normal health signs. Pyocyanin was non-toxic to the shrimp hybrid cell line (PmLyO-Sf9) at the concentration required for its application in disease management (IC₅₀?=?419.26 mg L^?1). The present study has demonstrated that pyocyanin is effective as an environment-friendly and safe aquaculture drug for the application in RAS-based shrimp culture to control Vibrio spp. without impeding nitrification established through the deployment of nitrifying bioreactors.

     

Marine yeast glucans confer better protection than that of baker's yeast in Penaeus monodon against white spot syndrome virus infection

The immunostimulatory property of glucan isolates from three marine yeasts (Debaryomyces hansenii S8, Debaryomyces hansenii S169 and Candida tropicalis S186) and one Baker's yeast (Saccharomyces cerevisiae S36) as examined for potential application as immunostimulants in Penaeus monodon postlarvae against White Spot Syndrome Virus (WSSV) infection. Structural characterization of the glucan component in the isolates by proton nuclear magnetic resonance (NMR) indicated similar structures containing (1‐3)‐linked anhydroglucose repeat units (AGRUs) in the backbone with (1‐6)‐linked AGRUs in side chains that are (1‐6)‐linked to the backbone AGRUs. Glucan from C. tropicalis (S186) with the highest molecular weight and the lowest level of branching supported maximum survival (69%) followed by the other two marine yeast (S169 and S8) glucans of 27% and 23% respectively while glucan from Baker's yeast, S. cerevisiae S36 with the lowest molecular weight and the highest level of branching exhibited poor survival (4%) in P. monodon post challenge WSSV. The present study showed that the glucan isolate from marine yeast with a higher molecular weight and a lower degree of branching acts as better immunostimulants in P. monodon postlarvae than did the glucan isolate from S. cerevisiae.     

Molecular characterization and expression analysis of two RING-between-RING (RBR) ubiquitin ligase orthologues from the Asian seabass (Lates calcarifer)

Post-translational modifications like ubiquitination determine the fate of many cellular proteins. There are evidences about the role of these ubiquitinated proteins in the antiviral immune response. E3 ubiquitin ligases like RBR (RING between RING) proteins play a crucial role in the regulation of immune signalling. In the present study, two RBR genes (RNF114 and RNF144A) from Asian seabass (Lates calcarifer) were cloned and characterized. Phylogenetic analysis reveals that these genes were closely clustered with their counterparts from teleost origin. Structurally they feature multiple domains with RING1 and RING2 domains and an IBR (in between RING) domain. The challenge studies with poly(I:C) and RGNNV virus gave evidence that both Lates RNF114 and RNF144A genes play a crucial role in antiviral immune response. Our studies suggest that RBR is an important gene in innate immune response and shed light on antiviral immune research in Lates calcarifer and its aquaculture.

     

Alkali insoluble glucan extracted from Acremonium diospyri is a more potent immunostimulant in the Indian White Shrimp, Fenneropenaeus indicus than alkali soluble glucan

Effect of an extraction method on the structure of glucan and its immunostimulatory response in Fenneropenaeus indicus was investigated. Here we extracted alkali insoluble glucan (AIG) and alkali soluble glucan (ASG) from a filamentous fungi Acremonium diospyri following alkali–acid hydrolysis and the sodium hypochlorite oxidation and dimethyl sulphoxide extraction method respectively. Structural analysis showed that 85% of glucan in AIG was a (1→3)-β- d -glucan and it increased the prophenoloxidase and reactive oxygen intermediate activity when administered to F. indicus . On the other hand, ASG, which contained 93% (1→3)-α-glucan, did not induce significant immune response in shrimp. Here we report that the difference in immunostimulatory potential between AIG and ASG is due to the difference in the percentage of (1→3)-β- d -glucans present in each preparation, which varies with the method of extraction employed. Also our observations suggest that glucan can be used as a potential immunostimulant to shrimp, provided it contains (1→3)-β- d -glucan as the major fraction.     

Comparative efficacy of MS-222 and benzocaine as anaesthetics under simulated transport conditions of a tropical ornamental fish Puntius filamentosus (Valenciennes)

There is a growing commercial interest in the fish, Puntius filamentosus , in the ornamental fish trade in India and elsewhere. The trade is, however, hampered by severe mortalities during transport of the fish owing to insufficient data available on the use of anaesthetics. To resolve this problem, we evaluated the efficacy of two anaesthetics, MS-222 and benzocaine, in sedating P. filamentosus in simulated transportation experiments and used stress response parameters such as cortisol and blood glucose levels to perform assessments. We observed that MS-222 at 40 mg L⁻¹ and benzocaine at 20 mg L⁻¹ were sufficient to induce sedation for 48 h. Above these concentrations, both the anaesthetics adversely affected the fish and resulted in mortalities. Both anaesthetics significantly lowered the blood cortisol and glucose levels compared with the unsedated controls. Importantly, the anaesthetics treatment significantly lowered the post-transport mortality in the fish. The results of the study show that MS-222 and benzocaine could be used as sedatives to alleviate transport-related stress in P. filamentosus to improve their post-transport survival and hence reduce economic loss.     

Marine yeast diet confers better protection than its cell wall component (1-3)-β-d-glucan as an immunostimulant in Fenneropenaeus indicus

A comparative study was performed to evaluate the immunostimulatory effect of yeast and yeast-derived glucan in white prawn Fenneropenaeus indicus (sub-adults of ∼20 gm). Feed with a whole cell biomass of marine yeast Candida sake S165 (CSY) at a concentration of 10% (w/w) and another feed with 0.2% glucan of C. sake S165 (CSG) were used in the study. Fenneropenaeus indicus were fed with these diets for 40 days and subsequently challenged with the white spot syndrome virus (WSSV). Haematological parameters such as the total haemocyte count, phenoloxidase activity, superoxide anion (O₂⁻) level, haemolymph peroxidase level and post-challenge survival against WSSV infection were determined to assess the immune status. In the present experiment, a higher immunity index and post-challenge survival were recorded in shrimps fed with the whole cell yeast diet. The better immunostimulatory performance of the whole cell yeast diet compared with the glucan diet could be attributed to the cellular constituents of yeast including the cell wall glucan, nucleotides, carotenoid pigments and vitamins. Here we observed that whole cell yeast performed better as an immunostimulant than the extracted cell wall glucans. Therefore, the use of yeast biomass in diets, rather than the yeast cell wall extract, glucan, would confer better protection against microbial infection besides reducing the cost of shrimp production.