Optical tweezers for probing the interactions of ZnO and Ag nanoparticles with E. coli.

The antibacterial effect of nanoparticles is mainly studied on the ensembles of the bacteria. In contrast, the optical tweezer technique allows the investigation of similar effects on individual bacterium. E. coli is a self-propelled micro-swimmer and ATP-driven active microorganism. In this work, an optical tweezer is employed to examine the mechanical properties of E. coli incubated with ZnO and Ag nanoparticles (NP) in the growth medium. ZnO and Ag NP with a concentration of 10 µg/ml were dispersed in growth medium during active log-growth phase of E. coli. This E. coli-NP incubation is further continued for 12 h. The E. coli after incubation for 2 h, 6 h and 12 h were separately studied by the optical tweezer for their mechanical property. The IR laser (λ = 975 nm; power = 100 mW) was used for trapping the individual cells and estimated trapping force, trapping stiffness and corner frequency. The optical trapping force on E. coli incubated in nanoparticle suspension shows linear decreases with incubation time. This work brings the importance of optical trapping force measurement in probing the antibacterial stress due to nanoparticles on the individual bacterium.

Characterization and cytotoxicity assessment of biosurfactant derived from Lactobacillus pentosus NCIM 2912.

Noteworthy properties of biosurfactant (BS) are fascinating scientific fraternity to explore them for food, medicinal, cosmetic, or pharmaceutical etc. applications. Newer products intended for pharmaceutical purposes are mandatory to go through pragmatic evaluation protocols. BS, being less cytotoxic, offers an ideal candidature for widespread applications in the healthcare sector. The goal of the current study was the isolation, physico-chemical characterization, and safety assessment of cell-associated biosurfactant (CABS) from Lactobacillus pentosus NCIM 2912. The culture was grown in a 3-L fermentor to produce CABS from the cell pellets through procedures like centrifugation, filtration, dialysis, column chromatography, and freeze-drying. Further, physical properties like surface tension (ST), critical micelle concentration (CMC), contact angle (CA), emulsification activity, stability of emulsion (height of emulsion, the extent of coalescence, and appearance), and ionic character of CABS were evaluated. Analytical characterization through TLC, FTIR, NMR, and GC-MS was carried out. The physico-chemical characterization revealed CABS as an anionic, multicomponent glycolipopeptide having a hydrophobic chain comprising butanoic acid (C4), decanoic acid (C10), undecanoic acid (C11), tridecanoic acid (C13), hexadecenoic acid (C16), and octadecanoic acid (C18). The oil-in-water (O/W) emulsions formed by CABS with various oils (olive, sesame, soybean, coconut) were stabilized up to the 7th day of storage and were analogous with polysorbate 80 (emulsifier/defoamer used in food industries). The O/W emulsions are quite stable at room temperature with no evidence of coalescence of droplets around 1 week. The cytotoxicity of CABS was evaluated through MTT (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide) assay. Cytotoxicity study performed on the human embryonic kidney (HEK 293), mouse fibroblast ATCC L929 and human epithelial type (HEP-2) cell lines recorded viability of 90.3 ± 0.1%, 99.2 ± 0.43, and 94.3 ± 0.2% respectively. The toxicity of the BS was comparable to that of the commercially used rhamnolipid sample. Thus, CABS derived from L. pentosus NCIM 2912 pose promising applications in the pharmaceutical, food industries acquiescently. The multifunctional potential of the incredibly versatile microbial product like BS from lactic acid bacteria (LAB) certainly contributes to wider avenues for varied industries.

Large tuning in the electrowetting behaviour on ferroelectric PVDF-HFP/Teflon AF bilayer.

Electrowetting (EW) response on a dielectric depends on its permittivity value, Young contact angle and voltage amplitude. We present a large change in EW contact angle, from 163° to 80°, on the bilayer dielectric made up of ferroelectric PVDF-HFP with a thin layer of fluoropolymer. The thickness values of both layers were separately optimized for high effective capacitance essential for the large EW response. It reveals that the bilayer with ~ 500 nm thick PVDF-HFP layer and ~ 50 nm thin layer of Teflon results in the maximum value of effective dielectric constant, ε ≈ 8. Besides this gain, dc-voltage EW response exhibits hysteresis mainly due to polarization in the ferroelectric layer such that, hysteretic offset voltage was found to depend on the applied voltage amplitude and thickness of the dielectrics. Finally, bilayer was subjected to ac-voltage EW in silicone oil for ambient temperature ranging from - 25 to 70 °C. The consistent EW response in this ambient without any degradation/delamination of polymer surface confirmed the durability of the bilayer on the transparent ITO electrodes. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10853-021-06308-z.

A novel fatty alkene from marine bacteria: A thermo stable biosurfactant and its applications.

In this study, a novel thermo stable biosurfactants, 1-Pentanonacontene (C95H190) a fatty alkene and 3-Hydroxy-16-methylheptadecanoic acid (C18H36O3) were isolated from a marine isolate SGD-AC-13. Biosurfactants were produced using 1% yeast extract in tap water as production medium at 24 h in flask and 12 h in bioreactor. Using 16S rRNA gene sequence (1515 bp) and BCL card (bioMérieux VITEK®), strain was identified as Bacillus sp. Crude biosurfactant reduced the surface tension of distilled water to 31.32 ±â€¯0.93 mN/m with CMC value of 0.3 mg/ml. Cell free supernatant showed excellent emulsification and oil displacement activity with stability up to 160 °C, pH 6-12 and 50 g/L NaCl conc. Biosurfactants were characterized using FTIR, TLC, HPLC LC-MS and NMR spectroscopy. Cell free supernatant reduced the contact angle of distilled water droplet from 117° to 52.28° and of 2% pesticide from 78.77° to 73.42° while 750 µg/ml of crude biosurfactant reduced from 66.06° to 56.33° for 2% pesticide and recovered 35% ULO and 12% HWCO from the contaminated sand. To our best of knowledge, this is the first report of thermo stable fatty alkene as a biosurfactant and is structurally different from previously reported, with having potential application in agriculture, oil recovery and bioremediation.

Inhibition of pathogenic bacterial biofilms on PDMS based implants by L. acidophilus derived biosurfactant.

BACKGROUND: Lactobacillus spp. predominantly shows its presence as a normal mucosal flora of the mouth and intestine. Therefore, the objective of our research is to investigate the in-vitro conditions for the prospective of medically valuable biosurfactants (BSs) derived from Lactobacillus spp. Biosurfactant (BS) obtained from Lactobacillus spp. exhibit antibiofilm and antiadhesive activity against broad range of microbes. In the present study we investigated the production, purification and properties of key components of the cell-associated-biosurfactant (CABS) from Lactobacillus acidophilus NCIM 2903. RESULTS: Extracted, purified, freeze-dried CABS shows reduction in surface tension (SFT) of phosphate buffer saline (PBS @pH 7.0) from 71 to 26 mN/m and had a critical micelle concentration (CMC) of 23.6 mg/mL. The CABS showed reduction in interfacial tension (IFT) against various hydrocarbons and had effective spreading capability as reflected through the decrease in contact angle (CA) on different surfaces (polydimethylsiloxane - PDMS, Teflon tape, glass surface, polystyrene film and OHP sheet). The anionic nature of CABS displayed stability at different pH and temperatures and formed stable emulsions. Thin layer chromatography (TLC) and Fourier transform infrared spectroscopy (FTIR) revealed CABS as glycolipoprotein type. The Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE) showed presence of multiple bands in a molecular range of 14.4 to 60 kDa, with prominent bands of 45 kDa. The CABS has significant antiadhesion and antibiofilm activity against tested bacterial strains. CONCLUSION: The current challenging situation is to develop methods or search for the molecules that will prevent the formations of biofilm on medical bioimplants of PDMS based materials. These findings are supportive for the use of Lactobacilli derived BS as potential antiadhesive agent on various surfaces of biomedical devices.

Spontaneous electrification of fluoropolymer-water interfaces probed by electrowetting.

Fluoropolymers are widely used as coatings for their robustness, water-repellence, and chemical inertness. In contact with water, they are known to assume a negative surface charge, which is commonly attributed to adsorbed hydroxyl ions. Here, we demonstrate that a small fraction of these ions permanently sticks to surfaces of Teflon AF and Cytop, two of the most common fluoropolymer materials, upon prolonged exposure to water. Electrowetting measurements carried out after aging in water are used to quantify the density of 'trapped' charge. Values up to -0.07 and -0.2 mC m-2 are found for Teflon AF and for Cytop, respectively, at elevated pH. A similar charge trapping process is also observed upon aging in various non-aqueous polar liquids and in humid air. A careful analysis highlights the complementary nature of electrowetting and streaming potential measurements in quantifying interfacial energy and charge density. We discuss the possible mechanism of charge trapping and highlight the relevance of molecular scale processes for the long term stability and performance of fluoropolymer materials for applications in electrowetting and elsewhere.

Low Voltage Electrowetting on Ferroelectric PVDF-HFP Insulator with Highly Tunable Contact Angle Range.

We demonstrate a consistent electrowetting response on ferroelectric poly(vinylidene fluoride-co-hexafluoropropylene) (PVDF-HFP) insulator covered with a thin Teflon AF layer. This bilayer exhibits a factor of 3 enhancement in the contact angle modulation compared to that of conventional single-layered Teflon AF dielectric. On the basis of the proposed model the enhancement is attributed to the high value of effective dielectric constant (εeff ≈ 6) of the bilayer. Furthermore, the bilayer dielectric exhibits a hysteresis-free contact angle modulation over many AC voltage cycles. But the contact angle modulation for DC voltage shows a hysteresis because of the field-induced residual polarization in the ferroelectric layer. Finally, we show that a thin bilayer exhibits contact angle modulation of Δθ (U) ≈ 60° at merely 15 V amplitude of AC voltage indicating a potential dielectric for practical low voltage electrowetting applications. A proof of concept confirms electrowetting based rapid mixing of a fluorescent dye in aqueous glycerol solution for 15 V AC signal.

Biosurfactant/s from Lactobacilli species: Properties, challenges and potential biomedical applications.

Lactic acid bacteria are generally believed to have positive roles in maintaining good health and immune system in humans. A number of Lactobacilli spp. are known to produce important metabolites, among which biosurfactants in particular have shown antimicrobial activity against several pathogens in the intestinal tract and female urogenital tract partly through interfering with biofilm formation and adhesion to the epithelial cells surfaces. Around 46 reports are documented on biosurfactant production from Lactobacillus spp. of which six can be broadly classified as cell free biosurfactant and 40 as cell associated biosurfactants and only approximately 50% of those have reported on the structural composition which, in order of occurrence were mainly proteinaceous, glycolipidic, glycoproteins, or glycolipopeptides in nature. Due to the proteinaceous nature, most biosurfactant produced by strains of Lactobacillus are generally believed to be surlactin type with high potential toward impeding pathogens adherence. Researchers have recently focused on the anti-adhesive and antibiofilm properties of Lactobacilli-derived biosurfactants. This review briefly discusses the significance of Lactobacilli-derived biosurfactants and their potential applications in various fields. In addition, we highlight the exceptional prospects and challenges in fermentation economics of Lactobacillus spp.-derived biosurfactants' production processes.

Multiple Roles of Biosurfactants in Biofilms.

Microbial growth and biofilms formation are a continuous source of contamination on most surfaces with biological, inanimate, natural or man-made. The use of chemical surfactants in daily practice to control growth, presence or adhesion of microorganisms and ultimately the formation of biofilms and biofouling is therefore becoming essential. Synthetic surfactants are, however, not preferred or ideal and biologically derived surface active biosurfactants (BSs) molecules produced mainly by microorganisms are therefore becoming attractive and sought by many industries. The search for innovative and interesting BS molecules that have effective antimicrobial activities and to use as innovative alternatives to chemical surfactants with added antimicrobial value among many other advantages has been ongoing for some time. This review discusses the various roles of BS molecules in association with biofilm formation. Recent updates on several mechanisms involved in biofilm development and control are presented vide this article.

Why patchy diffusion-limited aggregation belongs to the directed-percolation universality class.

We present a possible link between nonequilibrium phase transition observed in patchy diffusion-limited aggregation (DLA) [M. J. Kartha and A. Sayeed, Phys. Lett. A 380, 2791 (2016)10.1016/j.physleta.2016.06.036] and directed bond percolation (DP). A system of directed percolation with patchy particles (patchy DP) in which the bond connectivity is established depending on patch size p is analyzed. It is observed that patchy DP starting from a single seed shows a nonequilibrium phase transition. Below a critical value of the patch size p_{c}, the system reaches an absorbing state above which is a fluctuating active state as observed in the DP system. The value of this nonuniversal parameter p_{c} is observed to be slightly higher than the value observed in patchy DLA. Close to the critical value, the order parameter P(∞)∼(p-p_{c})^{ß} where ß=0.272±0.010, which is consistent with the directed-percolation universality class. Therefore the intrinsic nature of patchy DP is responsible for the phase transition in patchy DLA. This study reveals that the estimated critical value of patch size p_{c}=0.80625±0.00020 in patchy DP is different from the critical bond probability p_{c}=0.6447 in the DP system. This elucidates that the bond probability in DP is not equivalent to the patch probability of a particular site. Our work also gives an insight into the problem related with formation of an extended network of pentagon subunits in connection with the virus capsid.

Hysteretic DC electrowetting by field-induced nano-structurations on polystyrene films.

Electrowetting (EW) offers executive wetting control of conductive liquids on several polymer surfaces. We report a peculiar electrowetting response for aqueous drops on a polystyrene (PS) dielectric surface in the presence of silicone oil. After the first direct current (DC) voltage cycle, the droplet failed to regain Young's angle, yielding contact angle hysteresis, which is close to a value found in ambient air. We conjecture that the hysteretic EW response appears from in situ surface modification using electric field induced water-ion contact with PS surface inducing nano-structuration by electro-hydrodynamic (EHD) instability. Atomic force microscopy confirms the formation of nano-structuration on the electrowetted surface. The effects of molecular weight, applied electric field, water conductivity and pH on nano-structuration are studied. Finally, the EW based nano-structuration on PS surface is used for the enhanced loading of aqueous dyes on hydrophobic surfaces.

Use of electrowetting to measure dynamic interfacial tensions of a microdrop.

The adsorption of surface active species to liquid-liquid and to solid-liquid interfaces can have dramatic effects in microfluidics. In this paper we show how electrowetting on dielectric can be used to monitor a dynamic liquid-liquid interfacial tension (IFT) with a time resolution of O(1 s) using amplitude modulation of the AC voltage. This straightforward method, which requires less than a microlitre of sample, is demonstrated for aqueous drops containing Triton X-100 surfactant on a Teflon AF-coated substrate and with heptane as the immiscible oil ambient. Under these conditions, next to extracting the oil-water IFT (γ(ow)), also the effective water-substrate IFT difference (Δγ(ws)) can be obtained from the oil-water IFT and the Young's angle. Both γ(ow) and γ(ws) decrease over time due to adsorption. The measured dynamic oil-water IFT compares well to results of pendant drop experiments.

Methods for investigating biosurfactants and bioemulsifiers: a review.

Microorganisms produce biosurfactant (BS)/bioemulsifier (BE) with wide structural and functional diversity which consequently results in the adoption of different techniques to investigate these diverse amphiphilic molecules. This review aims to compile information on different microbial screening methods, surface active products extraction procedures, and analytical terminologies used in this field. Different methods for screening microbial culture broth or cell biomass for surface active compounds production are also presented and their possible advantages and disadvantages highlighted. In addition, the most common methods for purification, detection, and structure determination for a wide range of BS and BE are introduced. Simple techniques such as precipitation using acetone, ammonium sulphate, solvent extraction, ultrafiltration, ion exchange, dialysis, ultrafiltration, lyophilization, isoelectric focusing (IEF), and thin layer chromatography (TLC) are described. Other more elaborate techniques including high pressure liquid chromatography (HPLC), infra red (IR), gas chromatography-mass spectroscopy (GC-MS), nuclear magnetic resonance (NMR), and fast atom bombardment mass spectroscopy (FAB-MS), protein digestion and amino acid sequencing are also elucidated. Various experimental strategies including static light scattering and hydrodynamic characterization for micelles have been discussed. A combination of various analytical methods are often essential in this area of research and a numbers of trials and errors to isolate, purify and characterize various surface active agents are required. This review introduces the various methodologies that are indispensable for studying biosurfactants and bioemulsifiers.

Biosurfactants, bioemulsifiers and exopolysaccharides from marine microorganisms.

Marine biosphere offers wealthy flora and fauna, which represents a vast natural resource of imperative functional commercial grade products. Among the various bioactive compounds, biosurfactant (BS)/bioemulsifiers (BE) are attracting major interest and attention due to their structural and functional diversity. The versatile properties of surface active molecules find numerous applications in various industries. Marine microorganisms such as Acinetobacter, Arthrobacter, Pseudomonas, Halomonas, Myroides, Corynebacteria, Bacillus, Alteromonas sp. have been studied for production of BS/BE and exopolysaccharides (EPS). Due to the enormity of marine biosphere, most of the marine microbial world remains unexplored. The discovery of potent BS/BE producing marine microorganism would enhance the use of environmental biodegradable surface active molecule and hopefully reduce total dependence or number of new application oriented towards the chemical synthetic surfactant industry. Our present review gives comprehensive information on BS/BE which has been reported to be produced by marine microorganisms and their possible potential future applications.

Hydrodynamic resistance of single confined moving drops in rectangular microchannels.

We integrate a sensitive microfluidic comparator into a T-junction device and report measurements of the excess pressure drop due to a single moving droplet confined in a rectangular microchannel. We specifically focus on drops that are not coated with surfactants and study the effects of drop size, droplet viscosity and capillary number on their hydrodynamic resistance. In the capillary number range of approximately 10(-3)-10(-2), we find two distinct regimes for hydrodynamic resistance behavior based on drop size. In regime I associated with small drops (drop length/channel width approximately <4), we find that the pressure drop is independent of the drop size and the capillary number, and depends weakly on the ratio of the viscosities of the two immiscible phases. In regime II, associated with large drops (drop length/channel width > approximately 4), depending on the viscosity ratio of the two phases, the hydrodynamic resistance could increase, decrease or remain unchanged with drop size. We present a simple model that qualitatively captures these experimental trends. This model reveals that the pressure drop in regime I is dominated by the dissipation due to the end caps, and in regime II by both the end caps and the central body of the droplet. Such fundamental understanding will enable the design of large-scale energy-efficient fluidic circuits by minimizing the overall pressure drop in a network and may also provide insights into controlling droplet traffic to build functional passively-driven two-phase microfluidic technologies.

Electrowetting-based microdrop tensiometer.

We performed electrowetting (EW) contact angle measurements to determine the interfacial tension between aqueous drops laden with various inorganic and organic solutes and various ambient oils. Using low frequency AC voltage, we obtained interfacial tensions from 5 to 72 mJ/m 2, in close agreement with macroscopic tensiometry for drop volumes between 20 and 2000 nL. In addition to the conventional EW geometry, we demonstrate the possibility of performing "contact-less" measurements without any loss of accuracy using interdigitated coplanar electrodes.