Search

Increasing levels of salinity in coastal and freshwater environments as a result of rise in sea level is linked to climate change and it impacts fish growth adversely. Etroplus suratensis (Pearlspot) is the largest species among the indigenous cichlid which is essentially brackishwater fish that has become naturally acclimated to freshwater. Understanding the basis of tolerance and acclimatization to increased environmental salinity and freshwater condition can help the regional aqua-farmers to mitigate predicted impacts of climate change in that region. Technically the last decade contributed many methodologies to understand the salinity adaptation mechanisms; suppression subtractive hybridization (SSH) library generation being one of them. In this study we performed SSH technique to identify genes involved in salinity adaptation and osmoregulation in gills of E. suratensis, generating from fishes exposed to seawater (SW-36 ‰) as compared to freshwater (FW-0 ‰). Here, SSH was used to identify alterations in gene transcription of E. suratensis after exposure to 36‰. A random 105 clones were selected and sanger sequenced from the subtractive c-DNA libraries which gave a total of 80 gene fragment sequences.

Validation of microbial infection pathways in eukaryotic cells is challenging in the control of various infectious diseases. Semiconductor nanocrystals, also called quantum dots (QD), due to their exceptional brightness and photostability can be exploited in the long term monitoring of pathogens in host cells. However, the limited information about interactions of QDs and their bioconjugates with microorganisms confines the microbiological applications of QDs. Here we investigate the binding and toxicity of CdSe/ZnS QDs to the free-swimming marine pathogenic bacteria Vibrio harveyi using fluorescence microscopy, elastase assay, polyacrylamide gel electrophoresis (PAGE), and comet assay. The electrostatic binding of QDs to the cell surface has been found effective for the detection of the bacteria in aqueous solutions and bacteria-infected mammalian cells. The electrostatic binding is evaluated by the transient reversal of the cell surface charge contributed by macromolecules such as heparan sulfate proteoglycan (HSPG). Essentially, no fluorescence is detected for those bacteria treated with NiCl2 that reverses the cell surface charge. On the other hand, the efficiency of the cell surface to adsorb QDs remains intact even after treatment with elastase, which denatures the outer membrane proteins (Omps), suggesting HSPG-based binding of QD to cell surface and subsequently QDs are internalized. PAGE and comet assays show that the interactions of QDs with V. harveyi do not impart any cytotoxicity or genotoxicity. Further, we evaluate the integrity of adsorbed QDs for the detection of bacterial infection to mammalian cells by taking mouse fibroblast L929 as the model. Here, the stable fluorescence of QDs present in V. harveyi enables us for identifying the infected host cells. In short, the current study shows the potentials of for the detection of pathogens but without causing any toxic effects, which can be a promising method for not only the detection of the progression or regression of pathogenic infections but also phototherapy of microbial infections.

We propose antimicrobial photodynamic therapy (aPDT) as an alternative strategy to reduce the use of antibiotics in shrimp larviculture systems. The growth of a multiple antibiotic resistant Vibrio harveyi strain was effectively controlled by treating the cells with Rose Bengal and photosensitizing for 30 min using a halogen lamp. This resulted in the death of >50% of the cells within the first 10 min of exposure and the 50% reduction in the cell wall integrity after 30 min could be attributed to the destruction of outer membrane protein of V. harveyi by reactive oxygen intermediates produced during the photosensitization. Further, mesocosm experiments with V. harveyi and Artemia nauplii demonstrated that in 30 min, the aPDT could kill 78.9% and 91.2% of heterotrophic bacterial and Vibrio population respectively. In conclusion, the study demonstrated that aPDT with its rapid action and as yet unreported resistance development possibilities could be a propitious strategy to reduce the use of antibiotics in shrimp larviculture systems and thereby, avoid their hazardous effects on human health and the ecosystem at large.