Planktonic and early-stage biofilm microbiota respond contrastingly to thermal discharge-created seawater warming.

Thermal-discharges from power plants highly disturb the biological communities of the receiving water body and understanding their influence is critical, given the relevance to global warming. We employed 16 S rRNA gene sequencing to examine the response of two dominant marine bacterial lifestyles (planktonic and biofilm) against elevated seawater temperature (+5 â„ƒ). Obtained results demonstrated that warming prompted high heterogeneity in diversity and composition of planktonic and biofilm microbiota, albeit both communities responded contrastingly. Alpha diversity revealed that temperature exhibited positive effect on biofilm microbiota and negative effect on planktonic microbiota. The community composition of planktonic microbiota shifted significantly in warming area, with decreased abundances of Bacteroidetes, Cyanobacteria, and Actinobacteria. Contrastingly, these bacterial groups exhibited opposite trend in biofilm microbiota. Co-occurrence networks of biofilm microbiota displayed higher node diversity and co-presence in warming area. The study concludes that with increasing ocean warming, marine biofilms and biofouling management strategies will be more challenging.

3-Hydroxy coumarin demonstrates anti-biofilm and anti-hyphal efficacy against Candida albicans via inhibition of cell-adhesion, morphogenesis, and virulent genes regulation.

Candida albicans, a common fungus of human flora, can become an opportunistic pathogen and causes invasive candidiasis in immunocompromised individuals. Biofilm formation is the prime cause of antibiotic resistance during C. albicans infections and treating biofilm-forming cells is challenging due to their intractable and persistent nature. The study intends to explore the therapeutic potential of naturally produced compounds by competitive marine bacteria residing in marine biofilms against C. albicans biofilm. To this end, 3-hydroxy coumarin (3HC), a compound identified from the cell-free culture supernatant of the marine bacterium Brevundimonas abyssalis, was found to exhibit anti-biofilm and anti-hyphal activity against both reference and clinical isolates of C. albicans. The compound demonstrated significant inhibitory effects on biofilms and impaired the yeast-to-hyphal transition, wrinkle, and filament morphology at the minimal biofilm inhibitory concentration (MBIC) of 250 µg mL-1. Intriguingly, quantitative PCR analysis of 3HC-treated C. albicans biofilm revealed significant downregulation of virulence genes (hst7, ume6, efg1, cph1, ras1, als1) associated with adhesion and morphogenesis. Moreover, 3HC displayed non-fungicidal and non-toxic characteristics against human erythrocytes and buccal cells. In conclusion, this study showed that marine biofilms are a hidden source of diverse therapeutic drugs, and 3HC could be a potent drug to treat C. albicans infections.

Sequestration of cobalt and nickel by biofilm forming bacteria isolated from spent nuclear fuel pool water.

In the current study, six bacterial types, isolated from spent nuclear fuel (SNF) pool facility, were investigated for their ability to sequester heavy metals (cobalt and nickel). Biofilm formation by the six bacterial isolates, viz., Bacillus subtilis, Staphylococcus species, Staphylococcus arlettae, Staphylococcus epidermidis, Staphylococcus auricularis, and Chryseobacterium gleum, were assayed, and they were found to have significant biofilm forming property. Their biofilms were characterised using confocal scanning laser microscopy, and their potential to accumulate Co2+ and Ni2+ from bulk solutions was analysed with respect to time. A comparative assessment of bioaccumulation capacity was done using biofilms, planktonic cells, and live vs dead cells. The strains accumulated Co2+ and Ni2+ in the range of 4 × 10-4 to 1 × 10-5 g/mg of cell biomass. It is interesting to note that dead biomass also showed significant removal of the two metal ions, suggesting an alternative process for metal removal. This study suggests that hostile environments can be a repertoire of putative bacterial species with potential heavy metals and other contaminants remediation properties.

Biofilm formation by sulphate-reducing bacteria on different metals and their prospective role in titanium corrosion.

This study describes the biofilm formation by sulphate-reducing bacteria (SRB) on different materials, which has implications for the biomedical, pharmaceutical, food and chemical process industries. SRB was chosen as a model organism being an anaerobic bacterium. Biofilm formation on different materials and corrosion of titanium by SRB were monitored with time using confocal laser scanning microscopy and fluorescent FISH probes were used to authenticate the SRB strain. The thickness of the mono-culture SRB biofilm has ranged from 4 to 24 µm during thed 12-84 hr; however, the maximum biofilm thickness (24 µm) was recorded after 60 hr of growth. Planktonic growth of the SRB strain showed a log phase up to 48 hr and the sulphide production ranged from 2 to 14 mg l-1. For a comparative account, the SRB biofilm formation on copper was chosen as a positive control. Finally, the putative role of extracellular electron transfer by SRB in the biocorrosion process and the plausible mechanism of pitting corrosion of titanium is described in detail.

Exploring the effects of organic loading rate and domestic wastewater treatment by algal-bacterial granules under natural daylight conditions.

Algal-bacterial granules or phototrophic granules (PGs) comprising phototrophic microorganisms and bacteria are explored in wastewater treatment for achieving both environmental and economic sustainability. This study describes development of PGs and their use in biological treatment of synthetic and real domestic wastewater (sewage) under natural daylight conditions and low organic loading rate (OLR). Development of PGs was sequentially recorded in a photobioreactor operated in photo-sequencing batch reactor (photo-SBR) mode at a low OLR of 1 kgCOD.m-3 .day-1 and the developed PGs was evaluated for treating synthetic wastewater and real municipal wastewater with 0.14 kg COD m-3 .day-1 . PGs formed in the photo-sequential batch reactor (SBR) were compact and dense and exhibited excellent settling properties. The removal efficiencies were determined to be up to 95%, 93%, 97%, 72%, and 88% for turbidity, COD, TOC, NH4 + -N, and NO2 - -N/NO3 - -N, respectively. Additionally, a reduction in total viable bacterial counts and fecal coliform bacteria up to 1.7 × 103 and 7.8 × 102 cfu.mL-1 , respectively, during treatment of real municipal wastewater was achieved. This study demonstrated cultivation of algal-bacterial granules or PGs and their application for treating real municipal wastewater under natural daylight and tropical climate conditions. Further studies are needed on understanding interactions among phototrophic, autotrophic, and heterotrophic microorganisms of complex algal-bacterial consortium for emerging applications in bioremediation and wastewater treatment. PRACTITIONER POINTS: Phototrophic granules (PGs) were cultivated from algal consortium and activated sludge inoculum in photo sequencing batch reactors. Granular photobioreactor was operated at low OLR of 1 kgCOD.m3 .day-1 for developing well-settling algal-bacterial granules. PGs were stable and showed efficient biological treatment of synthetic wastewater and real sewage. Removals for turbidity, pathogens, and ammonium were at 95%, 3-log, and 72%, respectively, from real sewage.

The role of S-layer protein (SlpA) in biofilm-formation of Deinococcus radiodurans.

AIMS: To investigate the molecular basis of biofilm formation in a recombinant lab strain of Deinococcus radiodurans with a plasmid harbouring gfp and kanR that acquired the biofilm-forming ability. METHODS AND RESULTS: Deinococcus radiodurans R1 is known as a nonbiofilm former bacterium and so far there are no reports on its biofilm-producing capabilities. In this study, we investigated the molecular basis of biofilm formation in a recombinant strain of D. radiodurans using classical biofilm assays, confocal laser scanning microscopy and real-time PCR. Biochemical analysis of D. radiodurans biofilm matrix revealed that it consisted predominantly of protein and carbohydrate complexes with a little amount of extracellular DNA (eDNA). Furthermore, studies showed that D. radiodurans biofilm formation was enhanced in the presence of 25 mM Ca2+ , which enhanced the exopolysaccharide and protein content in the biofilm matrix. Enzymatic treatments with proteinase K, alginate lyase and DNase I indicated the involvement of some proteinaceous components to be critical in the biofilm formation. RT-PCR studies showed that increased expression of a surface layer protein SlpA conferred the biofilm ability to D. radiodurans. CONCLUSION: Overexpression of SlpA in D. radiodurans conferred the biofilm formation ability to the bacterium, in which a partial role was also played by the recombinant plasmid pKG. It was also shown that the presence of Ca2+ in the growth medium enhanced SlpA production, thus improving biofilm stability and biofilm maturation of D. radiodurans. SIGNIFICANCE AND IMPACT: This study shows how biofilm formation can be augmented in D. radiodurans. The finding has implications for the development of D. radiodurans biofilm-based biotechnological applications.

Copper oxide nanoparticles as an effective anti-biofilm agent against a copper tolerant marine bacterium, Staphylococcus lentus.

Biofilm formation on antifouling coatings is a serious concern in seawater cooling systems and the maritime industry. A prolific biofilm forming strain (Staphylococcus lentus), possessing high tolerance (>1,000 µg ml-1) to dissolved copper ions (Cu++) was isolated from titanium coupons exposed in the coastal waters of Kalpakkam, east coast of India. S. lentus formed increased biofilm (p < 0.05) at 100 µg ml-1 of Cu++ ions, when compared with the untreated control. To combat biofilm formation of this strain, the efficacy of copper oxide nanoparticles synthesized from copper nitrate by varying the concentrations of hexamine and cetyl trimethyl ammonium bromide (CTAB), was investigated. Complete (100%) inhibition of biofilm formation was observed with plain CuO NP (0.5 M hexamine, uncapped) at 1,000 µg ml-1. Capping with CTAB, influenced the morphology and the purity of the synthesized CuO NPs but did not alter their surface charge. Capping reduced metal ion release from CuO NPs and their antibacterial and anti-biofilm property against S. lentus. Overall, uncapped CuO NPs were effective in controlling biofilm formation of S. lentus. Concurrent release of copper ions and contact mediated physical damage by CuO NPs offer a promising approach to tackle metal tolerant biofilm bacteria.

Isolation and characterization of culturable bacteria present in the spent nuclear fuel pool water.

A spent nuclear fuel (SNF) pool is a key facility for safe management of nuclear waste, where spent nuclear fuel rods are stored in a water pool. The spent fuel rods carry a significant amount of radioactivity; they are either recycled or stored for further processing. Pool water acts as a heat sink as well as a shield against the radiation present in spent/burned fuel rods. The water used in these pools is filtered by an ultra-filtration process which makes certain the purity of water. As the life span of these pools is approximately 20 to 40 years, the maintenance of pure water is a big challenge. A number of researchers have shown the presence of bacterial communities in this ultrapure water. The bacterial types present in SNF pool water is of increasing interest for their potential bioremediation applications for radioactive waste. The present study showed the isolation of six bacterial species in the SNF pool water samples, which had significant radio-tolerance (D10 value 248 Gy to 2 kGy) and also biofilm-forming capabilities. These strains were also investigated for their heavy metal removal capacity. Maximum biofilm-mediated heavy metal (Co and Ni) removal (up to 3.8 µg/mg of biomass) was observed by three isolates (FPB1, FPB4, and FPB6). The ability of these bacterial isolates to survive in radioactive environments can be of great interest for remediation of radioactive contaminants.

Improvement in the corrosion resistance of carbon steel in acidic condition using naphthalen-2-ylnaphthalene-2-carboxammide inhibitor.

In the present investigation, the anti corrosive performance of synthesized Naphthalen-2-yl Naphthalene-2-Carboxammide (NNC) in 1 N HCl solution as corrosion inhibitor on carbon steel was studied at room temperature. Potentiodynamic polarization study revealed the mixed behavior of the inhibitor NNC along with inhibition efficiency. The inhibition efficiency of NNC increases with increasing inhibitor concentration. The electrochemical noise analysis (ENA) and electrochemical impedance spectroscopy (EIS) studies have showed that noise resistance (Rn) and charge transfer resistance (Rct) values were increased with addition of inhibitor concentrations. The synthesized and adsorbed bonds of inhibitor with the metal atom were characterized by ATR-FTIR spectroscopy. The adsorption isotherm explored that the adsorption of NNC obeyed Langmuir adsorption to bond with metal surface at carbon steel/HCl solution interface. The thermodynamic properties were calculated to discuss the adsorption mechanism of corrosion inhibition. Atomic force microscope (AFM) study showed a less corroded and roughness surface morphology, which is due to the formation of protective film layer on the surface. Quantum chemical parameters were calculated and correlated with respect to inhibitive performance of NNC. The study revealed that the adsorption nature of inhibitor has both physisorption and chemisorption phenomena.

Dispersal of Bap-mediated Staphylococcus aureus biofilm by proteinase K.

The dominant role of biofilm-associated protein (Bap) in Staphylococcus aureus biofilm development prompted us to investigate Bap as a potential target for proteinase-mediated biofilm dispersion. Biofilm assay in microtitre plates showed that proteinase K hampered the early adhesion of cells as well as biofilm development. Proteinase K treatment of 24- and 48-h-old biofilms showed enhanced dispersion of bap-positive S. aureus biofilm; however, proteinase K did not affect the bap-negative S. aureus biofilm. When antibiotics were used in combination with proteinase K, significant enhancement in antibiotic action was noticed against bap-positive S. aureus biofilm. This study establishes that antibiotics in combination with proteinase K can be used for controlling S. aureus biofilms in whose development Bap surface protein has a major role. We propose that Bap protein could be a potential target for therapeutic control of S. aureus infections (for example, bovine mastitis).

Characterization of biofilms in different clinical M serotypes of Streptococcus pyogenes.

Streptococcus pyogenes is a notorious human pathogen responsible for a wide array of infections. The ability of S. pyogenes to form biofilms is an innate property during the pathogenesis of invasive infections. From the eleven M serotypes tested: M56, M74, M100, M65, M89 and st38 formed dense biofilms in 48 h. The present study is the first of its kind to report about the biofilm formation in the serotypes M56, M65 M74 M100 and st38. XTT reduction assay of the biofilms showed decreased metabolic activity with increase in incubation time. The surface architecture of the biofilms when observed by scanning electron microscopy (SEM) revealed the microcolony formation. Confocal laser scanning microscopy (CLSM) was used to compare the surface topography and thickness of biofilms between the biofilm formers with and without the addition of glucose. Interestingly a non-biofilm former (st2147) was induced to form biofilms with the addition of glucose. On correlating the drug (erythromycin) resistance of the various M serotypes with their biofilm forming ability we noticed that erythromycin sensitive strains were found to be good biofilm formers. We also noticed that biofilm formation in S. pyogenes is independent of sil gene.