Microscopic and spectroscopic study of the corona formation and unfolding of human haemoglobin in presence of ZnO nanoparticles

The interaction of zinc oxide nanoparticles (ZnO NPs) with human haemoglobin (Hb) is studied for the biologically safe application of ZnO NPs in the human body. The Hb corona is formed around the ZnO nanoparticles, directly observed from high‐resolution transmission electron microscopy (HRTEM) images. Hb formed ‘hard corona' on the surface of ZnO NPs from an exponential association mechanism over a very short duration, as well as unfolding of Hb that occurred over a long lifetime. Dynamic light scattering measurements demonstrated that the ZnO NPs were completely covered by Hb with shell thickness of c. 6 nm that formed a ‘hard corona'. Zeta potential measurements represented that the ZnO NPs were fully covered by Hb molecules using an exponential association mechanism. Tryptophans (TRY), as well as heme‐porphyrin moieties of Hb, are the major binding sites for ZnO NPs. The nature of the interaction between ZnO NPs and Hb was analysed from the fluorescence quenching of TRYs. Electrostatic interaction, along with the hydrophobic interaction between ZnO NPs and Hb, is responsible for the conformational change in Hb due to increase in the percentage of β‐sheets together with a decrease in α‐helices.     

Deciphering the Probiotic Potential of Bacillus amyloliquefaciens COFCAU_P1 Isolated from the Intestine of Labeo rohita Through In Vitro and Genetic Assessment

Probiotics and Antimicrobial Proteins - In this study, a bacterial strain COFCAU_P1, isolated from the digestive tract of a freshwater teleost rohu (Labeo rohita), was identified as Bacillus...     

Resilience to Freezing in the Vegetative Cells of the Microalga Lobosphaera incisa (Trebouxiophyceae,Chlorophyta)

The chlorophyte microalga Lobosphaera incisa was isolated from the snowy slopes of Mt. Tateyama in Japan. This microalga stores exceptionally high amounts of the omega-6 LC-PUFA arachidonic acid in triacylglycerols, and therefore represents a potent photosynthetic source for this essential LC-PUFA. Assuming that freezing tolerance may play a role in adaptation of L. incisa to specific ecological niches, we examined the capability of L. incisa to tolerate extreme sub-zero temperatures. We report here, that the vegetative cells of L. incisa survived freezing at −20°C and −80°C (over 1 month), without cryoprotective agents or prior treatments. Cells successfully recovered upon thawing and proliferated under optimal growth conditions (25°C). However, cells frozen at −80°C showed better recovery and lower cellular ROS generation upon thawing, compared to those preserved at −20°C. Photosynthetic yield of PSII, estimated by F_v/F_m, temporarily decreased at day 1 post freezing and resumed to the original level at day 3. Interestingly, the thawed algal cultures produced a higher level of chlorophylls, exceeding the control culture. The polar metabolome of the vegetative cells comprised a range of compatible solutes, dominated by glutamate, sucrose, and proline. We posit that the presence of endogenous cryoprotectants, a rigid multilayer cell wall, the high LC-PUFA content in membrane lipids, and putative cold-responsive proteins may contribute to the retention of functionality upon recovery from the frozen state, and therefore for the survival under cryospheric conditions. From the applied perspective, this beneficial property holds promise for the cryopreservation of starter cultures for research and commercial purposes.     

Pseudomonas aeruginosa FARP72 Offers Protection Against Aeromonas hydrophila Infection in Labeo rohita

Use of probiotics as the biocontrol agent for disease prevention in aquaculture is gaining importance as an alternative to the indiscriminate use of antibiotics and other chemotherapeutics. In view of this trend, the probiotic properties of a potent antagonistic bacterium, Pseudomonas aeruginosa FARP₇₂, was characterized in terms of safety, antagonistic activities, in vitro immunomodulation, and in vivo disease resistance. Immunomodulatory activity was ascertained by measuring the production of intracellular superoxide anion, nitric oxide, total leukocyte peroxidase content, and the leukocyte proliferation in head kidney leukocytes. The bacterium isolated from the skin mucus of freshwater catfish Clarias batrachus was harmless to Labeo rohita. It showed inhibitory activity against Aeromonas caviae, A. hydrophila, Edwardsiella tarda, Pseudomonas putida, and Streptococcus agalactiae as revealed by cross and parallel streaking methods. Significantly higher superoxide anion and nitric oxide production, peroxidase content, and proliferative responses of leucocytes delineated the strains’ ability to interact with immune cells to activate the immune system in vitro. Significant growth inhibition of A. hydrophila from 1.55 × 10⁵ CFU/mL was observed when co-cultured with P. aeruginosa FARP₇₂ in phosphate-buffered saline (PBS) at levels ranging from 2.61 × 10⁷ to 2.61 × 10⁹ CFU/mL in 10 days. P. aeruginosa FARP₇₂ increased the survival rate of rohu fingerlings against pathogenic A. hydrophila challenge in biocontrol study in vivo as determined by cohabitation challenge. These results suggest that P. aeruginosa FARP₇₂ is a potential probiotic strain and can be used in aquaculture to improve the health status and disease resistance of fish.

     

Immune responses and protection in catla (Catla catla) vaccinated against epizootic ulcerative syndrome

Three different antigenic preparations from the epizootic ulcerative syndrome (EUS) pathogen Aphanomyces invadans were evaluated as vaccine candidate in catla (Catla catla). Anti-catla enzyme immunoconjugate was prepared after isolating catla immunoglobulin and raising hyperimmune sera against it, in rabbit. Three antigens namely, fungal extract (FE), fungal extract mixed with Freund’s incomplete adjuvant (FIA) in a 1:1 (v/v) ratio (FE + A) and extra cellular product (ECP) were prepared and three groups of catla were vaccinated intramuscularly with all these antigens (200 μg/fish). Different cellular and humoral immune responses were measured for the entire vaccinated and control group on 0th, 5th, 15th and 25th day post vaccination. Thirty days after the vaccination, the fish were challenged with an A. invadans zoospore dose of 1 × 10⁵ ml⁻¹ and mortality and relative percent of survival (RPS) were recorded. Study of cellular immunological parameters including antigen-specific leukocyte proliferation, antigen-specific nitric oxide production and superoxide anion production showed significantly higher (p < 0.05) values, in general, on 5th and 15th day post vaccination than the 0th day. Among all the antigenic groups, FE + A showed most significant response compared to the other groups. Among the humoral immune responses, lysozyme activity showed almost similar trend like cellular parameters. Anti-Aphanomyces antibody production was measured by enzyme-linked immunosorbent assay (ELISA) and it was increased with increasing days of vaccination in all the vaccinated groups with the highest observed on 25th day. Among the antigens, FE + A showed the highest antibody production following vaccination. The result of the homologous pathogen challenge study showed reduction in mortality in all the vaccinated groups. However, this reduction was not statistically significant (p > 0.05). Increased immune responses and protection have important implications with regard to the control of EUS by vaccination.     

Epizootic ulcerative syndrome (EUS) in fish: history and current status of understanding

Epizootic ulcerative syndrome (EUS) is one of the most important diseases affecting more than 100 species of wild and cultured finfish. EUS was first reported in farmed ayu (Plecoglossus altivelis) from Japan in 1971 and has since then spread across different countries of four continents including Asia, Australia, North America and Africa. The spread of the disease, especially in Asia–Pacific region and Africa has led to substantial damage to the fish resources and livelihood of the fish farmers. No reports are available confirming the outbreak of the disease from Europe and South America. The latest outbreak of EUS has been reported from Canada in a new susceptible species brown bullhead, Ameiurus nebulosus. It seems that the disease has potential to spread further, owing to the epizootic nature of the disease and broad susceptible host range. Considering the global importance of this disease, this review provides the current status of understanding about the etiology, process of diseases development, species affected, diagnostic methods as well as control and preventive measures, in light of the historical developments in those areas.     

Study of the Adsorption of Human Hemoglobin to Silver (Ag) Nanoparticle Surface for the Detection of the Unfolding of Hemoglobin

In the present report, we focused on the detail study of the optical properties and structural characterization of the Ag NPs for the nanobioconjugate analysis and detection of the conformational structural change of the Hb. The detail optical and structural analysis of Ag NPs has been studied from UV–Vis absorption, emission spectrum, XRD, and HRTEM study. The proteins/Hb are attached immediately onto Ag NPs surface when NPs touch the biological fluids, forming protein corona (PC), which gives their biological identity. The NPs-PC bioconjugate is, more specifically, the true identity of NPs in the physiological world. The adsorption of Hb with Ag NP surfaces has been studied by monitoring the soret band and tryptophan band of Hb. The dynamics of the Hb adsorption on the Ag NPs showed the time constant of surface binding t₁?=?5.79 min and 10.23 min and surface reorganization t₂?=?500 min and 251.75 min with the use of small and large concentrations of Ag NPs, respectively. The absorption peak shape and size around the wavelength, λ ≈ 406.2 nm of the bioconjugate has been examined by Gaussian and Lorentz curve fitting analysis. The bioconjugate along with the PC formation has been analyzed by HRTEM images and DLS observations. The tertiary deformation of Hb and energy transfer efficiency connecting Ag NPs and Hb are discussed from the emission-quenching phenomenon. The change of the secondary structural elements (α-helix, β-sheets, intermolecular aggregates, intramolecular aggregates) of the bioconjugate has been analyzed from FTIR spectrum.

     

In Vitro Assessment of Probiotic Potential of an Autochthonous Bacterial Isolate,Pseudomonas mosselii COFCAU_PMP5

Microbiology - Microbiota of the fish gut contains many beneficial bacteria that can be used as probiotics in aquaculture. In this study, several in vitro assays along with an in vivo safety...