Remarkable Improvement in Am3+ and Cm3+ Separation Using a Cooperative Counter Selectivity Strategy by a Combination of Branched Diglycolamides and Hydrophilic Polyaza-heterocycles.

Separation of Am3+ and Cm3+ is one of the most challenging problems in the back-end of the nuclear fuel cycle. In the present work, we exploited the cooperative effect of the opposite selectivity of hydrophobic branched DGA derivatives and hydrophobic N-donor heterocyclic ligands taken in two different phases to achieve improved separation behavior. A systematic study was performed using a series of DGA derivatives to understand the effect and the position of branching in the alkyl chains on the separation behavior of Am3+ and Cm3+. A separation factor (S.F.) value as high as 10 for Cm3+ over Am3+ was obtained in the case of TiBDGA (N,N,N',N'-tetra-iso-butyl diglycolamide) using SO3PhBTPhen ((phenanthroline-2,9-diyl)-1,2,4-triazine-5,5,6,6-tetrayltetrabenzenesulfonic acid) as the aqueous complexant, which is the highest reported value so far for the ligand-based separation of Am3+ and Cm3+ without involving any oxidation or reduction step. The high selectivity favoring Cm3+ ion extraction in the case of this DGA derivative is also explained with the help of computational studies.

Complete genome sequence of Halomonas boliviensis strain kknpp38, a chlorine-resistant bacterium isolated from the early-stage marine biofilm.

H. boliviensis strain kknpp38 is a dense exopolysaccharide (EPS) producing bacterium, isolated from the early-stage (72-h-old) of marine biofilm. Laboratory experiments demonstrated that this isolate forms a potent biofilm on various artificial substrata viz. polystyrene, stainless steel as well as titanium and possesses high tolerance to chlorine disinfection. To determine the genes and biosynthetic pathways involved in the EPS production, whole-genome sequencing was performed using high-throughput Illumina tag sequencing. The high-quality reads were first de novo assembled using Unicycler genome assembler (version 0.4.9b) and then annotated using Prokka (version 1.13). The complete genome comes from one circular chromosome containing 4.96 Mbp DNA with G + C content of 55%, and encompasses genes encoding 4476 proteins, 2 rRNAs, and 57 tRNAs. Intriguingly, genomic analysis revealed the existence of genes involved in ATP-binding cassette (ABC) transporter-dependent EPS biosynthesis pathways (ugd, ugd2, galU). In addition, we identified genes involved in ectoine (ectA, ectB, ectC, ectD) and polyhydroxyalkanoates (PHAs; fabA, fabB, fabD, fabF, fabH, fabV, fabZ, phaC, phaD, phaG, phaR, phaZ1) production, which are known to involve in bacterial adaptation in saline environment. The outcomes of this study expand scientific understanding on the genes and pathways involved in EPS biosynthesis by marine bacteria.

Bacterial community structure of early-stage biofilms is dictated by temporal succession rather than substrate types in the southern coastal seawater of India.

Bacterial communities colonized on submerged substrata are recognized as a key factor in the formation of complex biofouling phenomenon in the marine environment. Despite massive maritime activities and a large industrial sector in the nearshore of the Laccadive Sea, studies describing pioneer bacterial colonizers and community succession during the early-stage biofilm are scarce. We investigated the biofilm-forming bacterial community succession on three substrata viz. stainless steel, high-density polyethylene, and titanium over 15 days of immersion in the seawater intake area of a power plant, located in the southern coastal region of India. The bacterial community composition of biofilms and peripheral seawater were analyzed by Illumina MiSeq sequenced 16S rRNA gene amplicons. The obtained metataxonomic results indicated a profound influence of temporal succession over substrate type on the early-stage biofilm-forming microbiota. Bacterial communities showed vivid temporal dynamics that involved variations in abundant bacterial groups. The proportion of dominant phyla viz. Proteobacteria decreased over biofilm succession days, while Bacteroidetes increased, suggesting their role as initial and late colonizers, respectively. A rapid fluctuation in the proportion of two bacterial orders viz. Alteromonadales and Vibrionales were observed throughout the successional stages. LEfSe analysis identified specific bacterial groups at all stages of biofilm development, whereas no substrata type-specific groups were observed. Furthermore, the results of PCoA and UPGMA hierarchical clustering demonstrated that the biofilm-forming community varied considerably from the planktonic community. Phylum Proteobacteria preponderated the biofilm-forming community, while the Bacteroidetes, Cyanobacteria, and Actinobacteria dominated the planktonic community. Overall, our results refute the common assumption that substrate material has a decisive impact on biofilm formation; rather, it portrayed that the temporal succession overshadowed the influence of the substrate material. Our findings provide a scientific understanding of the factors shaping initial biofilm development in the marine environment and will help in designing efficient site-specific anti-biofouling strategies.

Draft Genome Sequencing of Pseudoalteromonas tetraodonis Strain kknpp56, a Potent Biofilm-Forming Bacterium Isolated from Early-Stage Marine Biofilm.

Pseudoalteromonas tetraodonis strain kknpp56 is an exopolysaccharide (EPS)-producing marine bacterium that forms potent biofilm. To determine the biosynthesis pathways involved in the EPS production of this bacterium, whole-genome sequencing was performed. The complete genome comes from one chromosome containing 3.72 Mbp of DNA with a G+C content of 41%.

Thermal discharge-induced seawater warming alters richness, community composition and interactions of bacterioplankton assemblages in a coastal ecosystem.

Despite accumulating evidence on the impact of global climate warming on marine microbes, how increasing seawater temperature influences the marine bacterioplankton communities is elusive. As temperature gradient created by thermal discharges provides a suitable in situ model to study the influence of warming on marine microorganisms, surface seawater were sampled consecutively for one year (September-2016 to August-2017) from the control (unimpacted) and thermal discharge-impacted areas of a coastal power plant, located in India. The bacterioplankton community differences between control (n = 16) and thermal discharge-impacted (n = 26) areas, as investigated using 16S rRNA gene tag sequencing revealed reduced richness and varied community composition at thermal discharge-impacted areas. The relative proportion of Proteobacteria was found to be higher (average ~ 15%) while, Bacteroidetes was lower (average ~ 10%) at thermal discharge-impacted areas. Intriguingly, thermal discharge-impacted areas were overrepresented by several potential pathogenic bacterial genera (e.g. Pseudomonas, Acinetobacter, Sulfitobacter, Vibrio) and other native marine genera (e.g. Marinobacter, Pseudoalteromonas, Alteromonas, Pseudidiomarina, Halomonas). Further, co-occurrence networks demonstrated that complexity and connectivity of networks were altered in warming condition. Altogether, results indicated that increasing temperature has a profound impact on marine bacterioplankton richness, community composition, and inter-species interactions. Our findings are immensely important in forecasting the consequences of future climate changes especially, ocean warming on marine microbiota.

Uranium sequestration abilities of Bacillus bacterium isolated from an alkaline mining region.

The present study elaborates uranium sequestration by bacteria from alkaline wastewaters. In the investigation, a few bacterial strains were isolated from alkaline uranium mine water and were tested for uranium sequestration properties 16S rRNA analysis assigned the 10 bacterial isolates to 4 genera of Actinobacteria and Firmicutes. Among all the isolates tested, the strain Bacillus aryabhattai (TP03) has shown superior sequestration capacity at 5 and 10 mg/L U in 1 mM carbonate-bicarbonate buffer at pH 9.2. At low uranium concentrations (5 mg/L as uranyl carbonate), the strain could sequester ~70% of the uranium in 6 h with a loading capacity of 4.3 mg U/g dry bacterial biomass. Increase in carbonate-bicarbonate buffer concentrations and pH reduced the sequestration capacity. Scanning electron microscopy and energy dispersive X-ray fluorescence spectroscopy studies indicated the presence of uranium with the bacterial biomass. Fourier transform infra-red spectroscopy results confirmed the uranium sequestration by cell membrane phosphate, amide, and carboxyl functional groups. Transmission electron microscopy study showed uranium presence within the cell cytoplasm, thus supporting the hypothesis on active metabolism-dependent bioaccumulation of uranium. The kinetics study of uranium sequestration was well fitted to the pseudo-second-order model. Overall, this study infers that the isolated alkaliphilic bacteria from the mine waters have significant sequestration property for treating uranium-containing alkaline wastewaters.

Exploring the impacts of heavy metals on spatial variations of sediment-associated bacterial communities.

One of the fundamental objectives in modern ecology is to decipher how bacterial communities in natural environment respond to anthropogenic activities. In recent times consequences of marine pollution, especially with heavy metals (HMs) have received increasing attention. However, insights into the response of bacterial communities to HMs in coastal sediments of India remain scarce. Here, we analyzed HMs content in three areas, along the southern coastal region of India. Based on the calculated pollution indices viz., enrichment factor (EF), contamination factor (CF), geo-accumulation index (Igeo) and sediment quality guidelines (SQGs), the studied areas were classified as uncontaminated, moderately contaminated and significantly contaminated. To explore the response of bacterial community to HMs, sediment-associated microbiota was investigated using high-throughput 16S rRNA gene amplicon sequencing. The obtained metataxonomic results revealed that bacterial diversity and community composition varied considerably in significantly contaminated area than moderately contaminated and uncontaminated areas. Proportion of bacterial classes was higher for Gammaproteobacteria, Betaproteobacteria and Actinobacteria, but lower for Alphaproteobacteria and Flavobacteriia in significantly contaminated area. Also, samples of significantly contaminated area were dominated by well-documented metal-resistant bacterial genera such as Ralstonia and Arthrobacter. Canonical correspondence analysis (CCA) showed that spatial variability of bacterial community composition was strongly correlated with HMs content such as Chromium, Cadmium and Nickel. Further analysis using PICRUSt programme indicated that the predictive functional profile also varied considerably in significantly contaminated area. By linking HMs with bacterial compositional variations, the present study highlights the likely influence of HMs in shaping sedimentary microbiota of coastal regions.

Sediment-associated bacterial community and predictive functionalities are influenced by choice of 16S ribosomal RNA hypervariable region(s): An amplicon-based diversity study.

Meta-omics approaches such as high-throughput sequencing of 16S hypervariable region(s) [HVR(s)] is extensively applied for profiling microbial community. Several studies have deciphered the influence of HVR(s) on bacterial diversity; most of these were devoted to human body habitats. Extent to which targeted HVR(s) influences the diversity estimates of environmental samples is rather unclear. Here, we evaluated the performance of five widely used universal primer pairs spanning V1-V3, V3-V4, V4, V5-V6 and V7-V9 HVRs to characterize bacterial diversity and predictive functionality of complex marine sediments. Obtained results revealed that the HVR(s) V4 and V5-V6 represented the higher species richness than others while, V1-V3 and V7-V9 were unsuccessful to detect Bacteroidetes and Planctomycetes. Further, PICRUSt analysis showed that the selected HVR(s) also had significant impact on the predictive functional profile. Conclusively, this study proved that HVR selection has a profound effect on overall results and thus should be selected with utmost caution.

Impediment to growth and yeast-to-hyphae transition in Candida albicans by copper oxide nanoparticles.

The effects of two prominent copper oxide nanoparticles (CuO-NP and Cu2O-NP), with the oxidation state of Cu++ (cupric) and Cu+ (cuprous), on Candida albicans were evaluated. CuO-NP and Cu2O-NP were synthesized and characterized by XRD, FESEM, HR-TEM and Zeta potential. At sub-MIC (50 µg ml-1), both cupric and cuprous oxide NPs prevented yeast-to-hyphae switching and wrinkling behaviour in C. albicans. The mechanism for the antifungal action of the two NPs differed; CuO-NP significantly elicited reactive oxygen species, whereas membrane damage was more pronounced with Cu2O-NP. Real time PCR analysis revealed that CuO-NP suppressed the morphological switching of yeast-to-hyphae by down-regulating cph1, hst7 and ras1 and by up-regulation of the negative regulator tup1. In comparison, Cu2O-NP resulted in down-regulation of ras1 and up-regulation of the negative regulators nrg1 and tup1. Between the two NPs, CuO exhibited increased antifungal activity due to its stable oxidation state (Cu++) and its smaller dimensions compared with Cu2O-NP.

Culture dependent and independent analysis and appraisal of early stage biofilm-forming bacterial community composition in the Southern coastal seawater of India.

Microbial aggregation on artificial surfaces is a fundamental phenomenon in aquatic systems that lead to biofouling, corrosion and influence the buoyancy of plastic materials. Despite the maritime activities and with nearshore large industrial sector, Laccadive Sea in the Indian Ocean has rarely been investigated for characterizing early biofilm-forming bacterial community. The present investigation was aimed to catalogue the primary colonizers on artificial surfaces and their comparison with planktonic community in southern coastal seawater of India. Surface seawater samples and biofilm assembled on three artificial surfaces over a period of 72 h of immersion in the intake area of a nuclear power plant at Kudankulam, India were collected. The structure of surface assemblages and plankton were unveiled by employing culture dependent, DGGE and NGS methods. In static condition, a collection of aerobic heterotrophic bacteria was screened in vitro for their ability to form potent biofilm. Proteobacteria preponderated the communities both in seawater and natural biofilm and Gammaproteobacteria accounted for >85% in the latter. Vibrionaceae, Alteromonadaceae and Pseudoalteromonadaceae dominated the biofilm community and constituted for 41, 25 and 8%, respectively. In contrast to other studies that showed Rhodobacteraceae family of Alphaproteobacteria as predominant component, we found Vibrionaceae of Gammaproteobacteria as dominant group in early stage of biofilm formation. Both DGGE and NGS data indicated that the attached community is noticeably distinct from those suspended in water column and form the basis for the proposed hypothesis of species sorting theory, that is, the local environmental conditions influence bacterial community assembly. Collectively, the data are testament for species sorting process that occur during initial assembly of bacterial community in marine environment and shed light on the structure of marine bacterial biofilm development. The outcome of the present study is of immense importance for designing long-term, efficient and appropriate strategies to control the biofouling phenomenon.

Characterization of microfouling and corrosive bacterial community of a firewater distribution system.

This investigation provides generic information on the culturable corrosive and the microfouling bacterial community in a firewater distribution system that uses freshwater. Conventional microbiological methods were used for the selective isolation of the major microfouling bacteria. The isolates were characterized by 16S rRNA gene sequencing and the biofilm as well as the corrosion characteristics of the isolates were evaluated. Pseudomonas aeruginosa and Bacillus cereus were predominantly observed in all the samples analysed. Denaturing gradient gel electrophoresis (DGGE) was carried out for the various samples of firewater system (FWS) and the high intensity bands were sequenced to identify the predominant bacteria. Bacterial groups such as Cyanobacteria, Proteobacteria, Actinobacteria, Bacteroidetes and Firmicutes were identified. Biofilm thickness was recorded using confocal scanning laser microscopy (CSLM). This was the first study to report Lysinibacillus fusiformis in a firewater system and its role in iron corrosion. Sulphidogenic bacteria Tissierella sp. and Clostridium bifermentans generated sulphides in the range of 400-900 ppm. Significant corrosion rates of carbon steel (CS) coupons were observed up to 4.3 mpy. C. bifermentans induced more localized corrosion in CS with a pit diameter of 50 µm. Overall, the data on the characterization of the fouling bacteria, their biofilm forming potential and subsequent metal deterioration studies supported in designing an effective water treatment program.