A Porphyrin-based NIR Fluorescent Probe for Bi3+ and Potential Applications.

Herein, we have prepared a 5,10,15,20-Tetrakis(4-hydroxyphenyl) porphyrin (P) which acts as a probe for selective and sensitive detection of Bi3+ ions. Probe P was obtained by reacting pyrrole with 4-hydroxyl benzaldehyde and characterized by NMR, IR, and ESI-MS. All photo-physical studies of P were tested in DMSO:H2O (8:2, v/v) media by spectrophotometry and spectrofluorometry respectively. The selectivity of P was tested with different metal ions in solution as well as in the solid phase, only Bi3+ showed red fluorescence quenching while with other metal ions, no such effect was observed. The Job's plot unveiled the 1:1 stoichiometric binding ratio of the probe with Bi3+ and anticipated association constant of 3.4 ×105 M-1, whereas the Stern-Volmer quenching constant was noticed to be 5.6 ×105 M-1. Probe P could detect Bi3+ down to 27 nM by spectrofluorometric. The binding mechanism of P with Bi3+ was well supported with NMR, mass, and DFT studies. Further, the P was applied for the quantitative determination of Bi3+ in various water samples and the biocompatibility of P was examined using neuro 2A (N2a) cells. Overall, probe P proves promising for the detection of Bi3+ in the semi-aqueous phase and it is the first report as a colorimetric and fluorogenic probe.

A Simple Live Cell Imaging "Turn-On" Fluorescence Probe for the Selective and Sensitive Detection of Aqueous Hg2+ Ions.

A simple, efficient, and reversible fluorescent sensor probe, PBA (2,6-dimethyl pyrone barbituric acid conjugate), comprised of a pro-aromatic donor conjugated with a barbituric acid, was developed for the detection of highly toxic mercuric ions. The probe showed high selectivity and "Turn-On" fluorescence response towards Hg2+ among various metal cations such as Na+, Mg2+, Ca2+, Mn2+, Fe2+, Co2+, Ni2+, Cu2+, Zn2+, Cd2+, Ba2+, Hg2+, and Pb2+, in both homogeneous and microheterogeneous micelle medium sodium dodecyl sulphate (SDS). The binding stoichiometry, limit of detection (LOD), and binding constant for the PBA-Hg complex were determined. The mechanism of binding was ascertained using the N,N'-dimethylbarbituric acid conjugate of 2,6-dimethylpyran (PDMBA), where no binding interaction by deprotonation is possible. In the presence of cysteamine hydrochloride and trifluoroacetic acid (TFA), the complexation of Hg2+ with PBA was demonstrated to be reversible, indicating its potential for the development of reusable sensors. Moreover, the practical applicability of PBA in monitoring Hg2+ in living cells was also evaluated.

Novel 3D porous aerogels engineered at nano scale from cellulose nano fibers and curcumin: An effective treatment for chronic wounds.

Traditional cotton gauze derived from cellulose has many limitations in the processes of wound healing. To overcome these hassles, we used cellulose nanofibers (CNF) incorporated with curcumin for the fabrication of wound healing 3D porous aerogel. Cellulose nanofibers synthesized from plant waste are promising sustainable nanomaterials due to their biocompatibility and biodegradability. Ionic cross linking with sodium alginate was performed to maintain the mechanical strength. SEM results revealed highly porous architecture that effectively promoted wound healing, as a result of macro- and micro-porous architecture and curcumin. In-vitro drug release studies showed a slow and steady release pattern. The 3D porous nano bio aerogel with curcumin significantly promoted the migration of fibroblast cells and had excellent antimicrobial activity against pathogenic microorganisms. In-vivo studies showed angiogenesis without rejection or inflammation of the scaffold. From the observations, we can conclude that this novel 3D porous aerogel can be used to treat chronic wounds.

Multi-functional bioactive secondary metabolites derived from endophytic fungi of marine algal origin.

Endophytic fungi from marine macroalgae are endowed with various pharmacologically active metabolites. This study mined, identified and screened endophytic fungi (EF) isolated from marine algae from the west coast of South India for screening anticancer, antioxidant and antimicrobial secondary metabolite producers. Five EF isolated from sampled marine algae were identified using morphology and ITS based identification as Grammothele fuligo, Rigidoporus vinctus, Cystobasidium minutum, Candida railenensis and Pichia kudriavzevii. After culturing 20 days on PDA medium, the ethyl acetate extracts of Cystobasidium minutum showed potent antimicrobial activity against P. aeruginosa with an IC50 value of 458.7 ± 1.021 µg/mL whereas Pichia kudriavzevii extracts showed promising antioxidant activity (IC50 value of 65.78 ± 1.082 µg/mL, 38.74 ± 1.040 µg/mL and 32.01 ± 1.018 µg/mL for DPPH assay, ABTS assay and FRAP assay respectively) and high cytotoxic activity against MG63 cell line (IC50 = 145.1 ± 1.086 µg/mL, no activity against U87 cells). The phytochemical screening of the extracts unveiled the existence of diverse groups of secondary metabolites. Further, Gas Chromatography Mass Spectroscopy (GC-MS) analysis of the extract revealed the presence of compounds that are known to be antibacterial, antioxidant and cytotoxic. These results indicate that marine derived endophytes could be potent sources for multi-functional bioactive compounds and may find prospective application in pharmaceutical industry.

Active bayerite underpinned Ag2O/Ag: an efficient antibacterial nanohybrid combating microbial contamination.

Active surfaces with bactericidal properties are of paramount importance in health care sector as a judicious approach to confront prevalent challenges presented by disastrous pathogenic infections and antibiotic-resistant microbes. Herein, we present Bayerite underpinned Ag2O/Ag (ALD), a nanohybrid with excellent antibacterial and antibiofilm functionalities against tested standard strains and clinical isolates. The multicomponent system coexists and complement each other with respect to phase and functionalities, demonstrated by XRD, XPS, and TEM analyses. In situ reduction of Ag+ ions to Ag0 over Bayerite as a stable bound phase is favoured by pH of the reaction, yielding 60-80% bound Ag protruding outwards facilitating active surface for interaction with microbes. ALD has a minimum inhibitory concentration (MIC) of 0.068 mg/ml against clinical isolates: Pseudomonas aeruginosa RRLP1, RRLP2, Acinetobactor baumannii C78 and C80. Disc diffusion assay demonstrated excellent antibacterial activity against standard strains (positive control: standard antibiotic disc, Amikacin). ALD incorporated PMMA films (5 and 10 wt%; PALD-5 and PALD-10) exhibited significant contact killing (99.9%) of clinical isolates in drop-test besides strong antibacterial activity (disc diffusion assay) comparable to that of ALD. ALD exemplified a dose (0.034 and 0.017 mg/ml) dependent biofilm inhibition (P < 0.001) and significant eradication of pre-formed biofilms (P < 0.001) by clinical isolates. PALD 5 and PALD 10 significantly declined the number of viable biofilm associated bacteria (99.9%) compared to control. Both ALD and PALD samples are proposed as green antibacterial materials with antibiofilm properties. Results also present ample opportunity to explore PALD as antibacterial and/or antibiofilm coating formulations.

In vitro efficiency evaluation of phage cocktail for biocontrol of Salmonella spp. in food products.

This study used a set of different bacteriophages to control contaminations of Salmonella spp., a major food pathogen. A cocktail of four phages designated based on host range and in vitro lytic assay showed a lower bacteriophage insensitive mutant frequency and considerable stability at 4 °C and 28 °C up to 60 days. The work evaluated the effectiveness of cocktail of four phages in reducing Salmonella spp. in four different food matrices (liquid egg, eggshell, milk, lettuce). A maximum of 1.7 log reduction in Salmonella spp. was achieved upon treatment of liquid eggs with phage cocktail for 72 h at 4 °C. In milk, the application of phage cocktail reduced recoverable Salmonella spp. by 1.9 log and 1.8 log at 28ºC (6 h) and 4ºC (72 h), respectively. A significant 2.9 log reduction of Salmonella spp. was obtained in eggshell after a 6 h incubation and Salmonella spp. was beyond detection level after 24 h at 28ºC. The application of cocktail also reduced Salmonella spp. beyond the detectable level in lettuce after 8 h at 28 °C. Our results indicated considerable stability of phages in different food matrices. Taken together, our findings establish the potential effectiveness of the bacteriophage cocktail as a biocontrol agent for Salmonella spp. in different food matrices.

Integration of heterogeneous photocatalysis and persulfate based oxidation using TiO2-reduced graphene oxide for water decontamination and disinfection.

Advanced oxidation processes (AOPs) which involve the generation of highly reactive free radicals have been considered as a promising technology for the decontamination of water from chemical and bacterial pollutants. In this study, integration of two major AOPs viz., heterogeneous photocatalysis involving TiO2-reduced graphene oxide (T-RGO) nanocomposite and activated persulfate (PS) based oxidation was attempted to remove diclofenac (DCF), a frequently detected pharmaceutical contaminant in water. The enhanced visible light responsiveness of T-RGO would facilitate the use of direct sunlight as a benign and cost effective source of energy for the photocatalytic activation. By combining PS based oxidation process with T-RGO mediated photocatalysis, a DCF removal efficiency of more than 98% was achieved within 30 min. The effect of operating parameters like PS concentration and pH on DCF removal was assessed. Radical scavenging experiments indicated that apart from radical oxidation involving •OH and SO 4 · - radicals, a non-radical oxidation pathway was also taking place in the degradation. The antibacterial properties of the integrated system were also evaluated using Escherichia coli and Staphylococcus aureus as representative bacteria. The presence of PS in the photocatalytic reaction system improved the antibacterial activity of the composite against the two strains studied. Cytotoxicity of T-RGO nanocomposite was assessed using human macrophage cell lines and the results showed that the composite is biocompatible and nontoxic at the recommended dosage for water treatment in the present study.

Optimization of PhysicoChemical Parameters for Production of Cytotoxic Secondary Metabolites and Apoptosis Induction Activities in the Culture Extract of a Marine Algal-Derived Endophytic Fungus Aspergillus sp.

The endophytic fungal community in the marine ecosystem has been demonstrated to be relevant source of novel and pharmacologically active secondary metabolites. The current study focused on the evaluation of cytotoxic and apoptosis induction potential in the culture extracts of endophytic fungi associated with Sargassum muticum, a marine brown alga. The cytotoxicity of the four marine endophytes, Aspergillus sp., Nigrospora sphaerica, Talaromyces purpureogenus, and Talaromyces stipitatus, was evaluated by the MTT assay on HeLa cells. Further, several physicochemical parameters, including growth curve, culture media, and organic solvents, were optimized for enhanced cytotoxic activity of the selected extract. The Aspergillus sp. ethyl acetate extract (ASE) showed maximum cytotoxicity on multiple cancer cell lines. Chemical investigation of the metabolites by gas chromatography-mass spectroscopy (GC-MS) showed the presence of several compounds, including quinoline, indole, 2,4-bis(1,1-dimethylethyl) phenol, and hexadecenoic acid, known to be cytotoxic in ASE. The ASE was then tested for cytotoxicity in vitro on a panel of six human cancer cell lines, namely, HeLa (cervical adenocarcinoma), MCF-7 (breast adenocarcinoma), Hep G2 (hepatocellular carcinoma), A-549 (lung carcinoma), A-431 (skin/epidermis carcinoma), and LN-229 (glioblastoma). HeLa cells were most vulnerable to ASE treatment with an IC50 value of 24 ± 2 µg/ml. The mechanism of cytotoxicity exhibited by the ASE was further investigated on Hela cells. The results showed that the ASE was capable of inducing apoptosis in HeLa cells through production of reactive oxygen species, depolarization of mitochondrial membrane, and activation of the caspase-3 pathway, which shows a possible activation of the intrinsic apoptosis pathway. It also arrested the HeLa cells at the G2/M phase of the cell cycle, eventually leading to apoptosis. Through this study, we add to the knowledge about the marine algae associated with fungal endophytes and report its potential for purifying specific compounds responsible for cytotoxicity.

Optical characterization and tunable antibacterial properties of gold nanoparticles with common proteins.

The interaction of three proteins, viz. Bovine Serum Albumin (BSA), Human Serum Albumin (HSA) and Hen Egg White Lysozyme (HEWL) with gold nanoparticles (GNPs) is investigated using surface plasmon resonance (SPR) spectroscopy, fluorescence spectroscopy and circular dichroism (CD). Size and morphology of the samples was established using Transmission Electron Microscopy (TEM) and stability studies was established using zeta potential analysis. The stability of protein-GNP complex was found to be greater than that of individual protein as well as individual GNPs. Also HEWL-GNP complex was more stable compared to the other protein complexes. Absorbance of proteins increases with increase in gold nanoparticle concentration due to the extension of peptide strands of protein and decrease in hydrophobicity of gold nanoparticles. A ground state complex is also formed which is evident from the moderate shift observed in the absorbance peaks. Apparent association constant was also determined from the absorption spectra and was found to be maximum for HEWL and minimum for HSA. Gold nanoparticles were found to act as quenchers and reduced the protein fluorescence intensity. Binding constant and number of binding sites were found to be maximum for HEWL and minimum for HSA. The temperature dependent fluorescence studies were also performed to calculate the thermodynamic parameters and to determine the nature of interaction between the proteins and gold nanoparticles. The circular dichroism studies elucidate the reason behind the maximum binding for HEWL and minimum binding for HSA. TGA analysis determined the thermal stability of the samples. Fluorescence lifetime studies indicate static quenching of proteins. Antibacterial activity of protein-gold nanoparticles was studied against four pathogens, viz. Bacillus pumilus, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. HEWL exhibits a tunable antimicrobial activity against Pseudomonas aeruginosa due to the maximum binding of HEWL with gold nanoparticles. The study proposes a novel method for adjusting the antibacterial activity of HEWL against Pseudomonas aeruginosa when the resistance of this pathogen is a major issue in the chemotherapy of many infectious diseases. Thus the combination therapy of protein-gold nanoparticles could prove to be a new approach in medical field in the near future.

Metagenomic data on bacterial diversity profiling of Arabian sea sediment by amplicon sequencing.

This data is about the microbial community genome analysis of Arabian sea sediment by Illumina sequencing by targeting the hypervariable region V3 of 16S rRNA gene. The data analysis revealed the existence of numerous unknown sequences, indicating a large unexploited bacterial diversity in the area. The raw sequence data used for analysis is available in NCBI under the Sequence Read Archive (SRA) with the BioProject No. PRJNA397165 and SRA accession number SRP125840.

Genome characterization of novel lytic Myoviridae bacteriophage ϕVP-1 enhances its applicability against MDR-biofilm-forming Vibrio parahaemolyticus.

A pathogen of significance in the aquaculture sector, the Gram-negative marine bacterium Vibrio parahaemolyticus causes gastroenteritis associated with consumption of improperly prepared seafood. This bacterium can be controlled using lytic bacteriophages as an alternative to antibiotics. Ï•VP-1 is a lytic phage of V. parahaemolyticus that was isolated from an aquafarm water sample with the aim of assessing its potential as a bio-control agent and determining its physicochemical properties and genomic sequence. Morphological analysis by transmission electron microscopy and phylogenetic analysis based on the large terminase subunit gene showed that this phage belongs to the family Myoviridae. It could infect multiple-drug-resistant (MDR) V. parahaemolyticus and V. alginolyticus strains of mangrove and seafood origin. With a maximum adsorption time of 30 min, ϕVP-1 has a short latent period of 10 min with burst size of 44 particles/cell. Whole-genome sequencing was done using the Illumina platform, and annotation was done using GeneMarkS and Prodigal. The 150,764bp genome with an overall G+C content of 41.84% had 203 putative protein-encoding open reading frames, one tRNA gene, and 66 predicted promoters. A number of putative DNA replication and regulation, DNA packaging and structure, and host lysis genes were identified. Comparison of the ϕVP-1 genome sequence to those of known Vibrio phages indicated little discernible DNA sequence similarity, suggesting that ϕVP-1 is a novel Vibrio phage. Sequence analysis revealed the presence of 64 potential ORFs with a T4-like genomic organization. In silico analysis suggested an obligate lytic life cycle and showed the absence of lysogeny or virulence genes. The complete sequence of Ï•VP-1 was annotated and deposited in the GenBank database (accession no. MH363700). The genetic features of this novel phage suggest that it might be applicable for phage therapy against pathogenic strains of V. parahaemolyticus.

Draft genome sequence data of T-5 like Salmonella bacteriophage ФSP3 with demonstrated therapeutic potential.

This data article describes the complete draft genome sequence of Salmonella specific bacteriophage ΦSP3 isolated from chicken intestinal contents. The ΦSP3 genome was sequenced by paired end runs using Illumina HiSeq 2500 with 100X coverage. Phylogenetic analysis using major capsid gene and genome wide comparison were performed to understand bacteriophage evolutionary relationship. Genome sequence of bacteriophage ΦSP3 was deposited in GenBank under the accession number MG387042.

Phage Endolysins as Potential Antimicrobials against Multidrug Resistant Vibrio alginolyticus and Vibrio parahaemolyticus: Current Status of Research and Challenges Ahead.

Vibrio alginolyticus and V. parahaemolyticus, the causative agents of Vibriosis in marine vertebrates and invertebrates, are also responsible for fatal illnesses such as gastroenteritis, septicemia, and necrotizing fasciitis in humans via the ingestion of contaminated seafood. Aquaculture farmers often rely on extensive prophylactic use of antibiotics in farmed fish to mitigate Vibrios and their biofilms. This has been postulated as being of serious concern in the escalation of antibiotic resistant Vibrios. For this reason, alternative strategies to combat aquaculture pathogens are in high demand. Bacteriophage-derived lytic enzymes and proteins are of interest to the scientific community as promising tools with which to diminish our dependency on antibiotics. Lysqdvp001 is the best-characterized endolysin with lytic activity against multiple species of Vibrios. Various homologues of Vibrio phage endolysins have also been studied for their antibacterial potential. These novel endolysins are the major focus of this mini review.

BaCf3: highly thermostable bacteriocin from Bacillus amyloliquefaciens BTSS3 antagonistic on food-borne pathogens.

In the present study, we characterized bacteriocin BaCf3, isolated and purified from Bacillus amyloliquefaciens BTSS3, and demonstrated its inhibitory potential on growth and biofilm formation of certain food spoilage bacteria and pathogens. Purification was by gel filtration chromatography and its molecular weight was 3028.422 Da after MALDI-TOF MS. The bacteriocin was highly thermostable withstanding even autoclaving conditions and pH tolerant (2.0-13.0). The bacteriocin was sensitive to oxidizing agent (DMSO) and reducing agent (DTT). The de novo sequence of the bacteriocin BaCf3 was identified and was found to be novel. The sequence analysis shows the presence of a disulphide linkage between C6 and C13. The microtitre plate assay proved that BaCf3 could reduce up to 80% biofilm produced by strong biofilm producers from food samples. In addition, BaCf3 did not show cytotoxicity on 3-TL3 cell line.

Data on the characterization of non-cytotoxic pyomelanin produced by marine Pseudomonas stutzeri BTCZ10 with cosmetological importance.

The article focuses on data dealing with characterization of black brown melanin produced by marine bacteria Pseudomonas stutzeri BTCZ10. Figures deal with the production of melanin by strain BTCZ10 and characterization of the pigment using biophysical techniques. Table presents the data on photo-protective ability of melanin when blended with commercial sunscreens.

Genomics of Salmonella phage ΦStp1: candidate bacteriophage for biocontrol.

Multidrug-resistant Salmonella causing Salmonellosis is a food-borne pathogen and hence a public health hazard. Alternatives to antibiotics, such as phages, are possible solutions to this increasing drug resistance. In this context, several Salmonella phages were isolated and characterized. This paper describes the physiochemical and whole genome characterization of one such bacteriophage, ΦStp1, which efficiently infects serovars Salmonella Enteritidis and Salmonella Typhimurium. Morphological observations by transmission electron microscopy and phylogenetic analysis using terminase gene classified ΦStp1 to family Siphoviridae, closely resembling 'T5 like phage' morpho-types. With a maximum adsorption time of 50 min, ΦStp1 latent period was 30 min with 37 phages/cell burst size. ΦStp1 draft genome sequenced by shotgun method comprised 112,149 bp in 3 contigs with 37.99% GC content, 168 predicted ORFs, and 15 tRNAs. Genes involved in host shut down, DNA replication, regulation, nucleotide metabolism, lysis, and morphogenesis were also noted. The study not only provided an insight into the characteristics of phage genome, but also information about proteins encoded by bacteriophages, therefore contributing to understanding phage diversity. Sequence analysis also proved the absence of virulence and lysogeny-related genes, which only went to confirm ΦStp1 as a promising therapeutic agent against Salmonella infections.

Isolation, purification and characterization of a pH tolerant and temperature stable proteinaceous protease inhibitor from marine Pseudomonas mendocina.

OBJECTIVES: An extracellular protease inhibitor (BTPI-301) of trypsin was purified and characterized from an isolate of Pseudomonas mendocina. RESULTS: BTPI-301was purified to homogeneity by (NH4)2SO4, precipitation, DEAE Sepharose and CNBr-activated Sepharose chromatography. Homogeneity was proved by native PAGE and SDS-PAGE. The intact molecular mass was 11567 Da by MALDI-TOF analysis. BTPI-301was a competitive inhibitor with a Ki of 3.5 × 10-10 M. It was stable and active at pH 4-12 and also at 4-90 °C for 1 h. Peptide mass fingerprinting by MALDI revealed that the BTPI-301 is a new inhibitor not reported so far with protease inhibitory activity. The pI of the inhibitor was 3.8. The stoichiometry of trypsin-BTPI-301 interaction is 1:1. The inhibitor was specific towards trypsin. CONCLUSION: A pH tolerant and thermostable protease inhibitor BTPI-301 active against trypsin was purified and characterized from P. mendocina that could be developed and used as biopreservative as well as biocontrol agent.

Molecular cloning and characterization of a halotolerant α-amylase from marine metagenomic library derived from Arabian Sea sediments.

Functional screening of a metagenomic library of marine sediment revealed an amylolytic clone BTM109. This report states the purification and characterization of a moderately halotolerant α-amylase, with more than 51% activity in 2.5 M NaCl. The molecular mass of purified protein was determined to be 55.7 kDa by MALDI-TOF MS. The optimum pH for enzyme activity was pH 7 and temperature for maximal activity was 40 °C. At 5 mM concentration, Ca2+ enhanced the enzyme activity indicating that the enzyme is a Ca2+ dependent α-amylase which was confirmed by the starch hydrolysis pattern using TLC. These physico-chemical properties support the suitability of this enzyme for various industrial applications.

Characterization of pyocyanin with radical scavenging and antibiofilm properties isolated from Pseudomonas aeruginosa strain BTRY1.

A biomolecule namely pyocyanin was isolated from Pseudomonas aeruginosa (BTRY1). It was confirmed by UV-Vis spectrum with absorption maxima at 270 nm and was further characterized using Fourier transform infrared (FTIR) and nuclear magnetic resonance (NMR). Spectroscopy analyses showed the presence of relevant bonds in their respective structures. Although pyocyanin had antihemolytic, antioxidant and antibiofilm activities against food pathogens, it showed no observable cytotoxicity, and thus proposed for potential use in the preservation of food.

Characterization of Deep Sea Fish Gut Bacteria with Antagonistic Potential, from Centroscyllium fabricii (Deep Sea Shark).

The bacterial isolates from Centroscyllium fabricii (deep sea shark) gut were screened for antagonistic activity by cross-streak method and disc diffusion assay. This study focuses on strain BTSS-3, which showed antimicrobial activity against pathogenic bacteria including Salmonella Typhimurium, Proteus vulgaris, Clostridium perfringens, Staphylococcus aureus, Bacillus cereus, Bacillus circulans, Bacillus macerans and Bacillus pumilus. BTSS3 was subjected to phenotypic characterization using biochemical tests, SEM imaging, exoenzyme profiling and antibiotic susceptibility tests. Comparative 16S rDNA gene sequence analysis indicated that this strain belonged to the genus Bacillus, with high (98%) similarity to 16S rDNA sequences of Bacillus amyloliquefaciens. The chemical nature of the antibacterial substance was identified by treatment with proteolytic enzymes. The antibacterial activity was reduced by the action of these enzymes pointing out its peptide nature. It was observed from the growth and production kinetics that the bacteriocin was produced in the eighth hour of incubation, i.e., during the mid-log growth phase of the bacteria.