Surface properties influence marine biofilm rheology, with implications for ship drag.

Marine biofilms on ship hulls increase frictional drag, which has economic and environmental consequences. It is hypothesised that biofilm mechanics, such as viscoelasticity, play a critical role in biofilm-associated drag, yet is a poorly studied area. The current study aimed to rheologically characterise ship-relevant marine biofilms. To combat marine biofilms on ship hulls, fouling-control coatings are often applied; therefore, the effect of different surfaces on marine biofilm mechanics was also investigated. Three surfaces were tested: a non-biocidal, chemically inert foul-release coating (FRC), an inert primer (ACP) and inert PVC. Physical properties of biofilms were explored using Optical Coherence Tomography (OCT) and a parallel-plate rheometer was used for rheological testing. Image analysis revealed differences in the thickness, roughness, and percent coverage between the different biofilms. Rheological testing showed that marine biofilms, grown on FRC and ACP acted as viscoelastic materials, although there were differences. FRC biofilms had a lower shear modulus, a higher viscosity, and a higher yield stress than the ACP biofilms, suggesting that the FRC biofilms were more readily deformable but potentially more robust. The results confirmed that surface treatment influences the structural and mechanical properties of ship-relevant marine biofilms, which could have implications for drag. A better understanding of how different surface treatments affect marine biofilm rheology is required to improve our knowledge on biofilm fluid-structure interactions and to better inform the coating industry of strategies to control biofilm formation and reduce drag.

Monitoring Aerobic Marine Bacterial Biofilms on Gold Electrode Surfaces and the Influence of Nitric Oxide Attachment Control.

Detection of aerobic marine bacterial biofilms using electrochemical impedance spectroscopy has been done to monitor the interfacial response of Pseudoalteromonas sp. NCIMB 2021 attachment and growth in order to identify characteristic events on a 0.2 mm diameter gold electrode surface. Uniquely, the applicability of surface charge density has been proven to be valuable in determining biofilm attachment and cell enumeration over a 72 h duration on a gold surface within a modified continuous culture flow cell (a controlled low laminar flow regime with Reynolds number ≈ 1). In addition, biofilm dispersal has been evaluated using 500 nM sodium nitroprusside, a nitric oxide donor (nitric oxide is important for the regulation of several diverse biological processes). Ex situ confocal microscopy studies have been performed to confirm biofilm coverage and morphology, plus the determination and quantification of the nitric oxide biofilm dispersal effects. Overall, the capability of the sensor to electrochemically detect the presence of initial bacterial biofilm formation and extent has been established and shown to have potential for real-time biofilm monitoring.

A rapid benchtop method to assess biofilm on marine fouling control coatings.

A rapid benchtop method to measure the torque associated with minidiscs rotating in water using a sensitive analytical rheometer has been used to monitor the drag caused by marine fouling on coated discs. The method was calibrated using sandpaper surfaces of known roughness. Minidiscs coated with commercial fouling control coatings, plus an inactive control, were exposed in an estuarine harbour. After 176 days the drag on the fouling control-coated discs, expressed as a moment coefficient, was between 73% and 90% less than the drag on the control coating. The method has potential use as a screen for novel antifouling and drag reducing coatings and surfaces. Roughness functions derived using Granville's indirect similarity law are similar to patterns found in the general hydrodynamics literature, and so rotational minidisc results can be considered with reference to other fouling drag datasets.Supplemental data for this article is available online at https://doi.org/10.1080/08927014.2021.1929937 .

Biofilm Inhibition by Novel Natural Product- and Biocide-Containing Coatings Using High-Throughput Screening.

The use of natural products (NPs) as possible alternative biocidal compounds for use in antifouling coatings has been the focus of research over the past decades. Despite the importance of this field, the efficacy of a given NP against biofilm (mainly bacteria and diatoms) formation is tested with the NP being in solution, while almost no studies test the effect of an NP once incorporated into a coating system. The development of a novel bioassay to assess the activity of NP-containing and biocide-containing coatings against marine biofilm formation has been achieved using a high-throughput microplate reader and highly sensitive confocal laser scanning microscopy (CLSM), as well as nucleic acid staining. Juglone, an isolated NP that has previously shown efficacy against bacterial attachment, was incorporated into a simple coating matrix. Biofilm formation over 48 h was assessed and compared against coatings containing the NP and the commonly used booster biocide, cuprous oxide. Leaching of the NP from the coating was quantified at two time points, 24 h and 48 h, showing evidence of both juglone and cuprous oxide being released. Results from the microplate reader showed that the NP coatings exhibited antifouling efficacy, significantly inhibiting biofilm formation when compared to the control coatings, while NP coatings and the cuprous oxide coatings performed equally well. CLSM results and COMSTAT analysis on biofilm 3D morphology showed comparable results when the NP coatings were tested against the controls, with higher biofilm biovolume and maximum thickness being found on the controls. This new method proved to be repeatable and insightful and we believe it is applicable in antifouling and other numerous applications where interactions between biofilm formation and surfaces is of interest.

Extracellular DNA impedes the transport of vancomycin in Staphylococcus epidermidis biofilms preexposed to subinhibitory concentrations of vancomycin.

Staphylococcus epidermidis biofilm formation is responsible for the persistence of orthopedic implant infections. Previous studies have shown that exposure of S. epidermidis biofilms to sub-MICs of antibiotics induced an increased level of biofilm persistence. BODIPY FL-vancomycin (a fluorescent vancomycin conjugate) and confocal microscopy were used to show that the penetration of vancomycin through sub-MIC-vancomycin-treated S. epidermidis biofilms was impeded compared to that of control, untreated biofilms. Further experiments showed an increase in the extracellular DNA (eDNA) concentration in biofilms preexposed to sub-MIC vancomycin, suggesting a potential role for eDNA in the hindrance of vancomycin activity. Exogenously added, S. epidermidis DNA increased the planktonic vancomycin MIC and protected biofilm cells from lethal vancomycin concentrations. Finally, isothermal titration calorimetry (ITC) revealed that the binding constant of DNA and vancomycin was 100-fold higher than the previously reported binding constant of vancomycin and its intended cellular d-Ala-d-Ala peptide target. This study provides an explanation of the eDNA-based mechanism of antibiotic tolerance in sub-MIC-vancomycin-treated S. epidermidis biofilms, which might be an important factor for the persistence of biofilm infections.

Marine biofilms on artificial surfaces: structure and dynamics.

The search for new antifouling (AF) coatings that are environmentally benign has led to renewed interest in the ways that micro-organisms colonize substrates in the marine environment. This review covers recently published research on the global species composition and dynamics of marine biofilms, consisting mainly of bacteria and diatoms found on man-made surfaces including AF coatings. Marine biofilms directly interact with larger organisms (macrofoulers) during colonization processes; hence, recent literature on understanding the basis of the biofilm/macrofouling interactions is essential and will also be reviewed here. Overall, differences have been identified in species composition between biofilm and planktonic forms for both diatoms and bacteria at various exposure sites. In most studies, the underlying biofilm was found to induce larval and spore settlement of macrofoulers; however, issues such as reproducibility, differences in exposure sites and biofilm composition (natural multispecies vs. monospecific species) may influence the outcomes.

Pseudotumour formation due to tribocorrosion at the taper interface of large diameter metal on polymer modular total hip replacements.

We present an in-depth failure analysis of two large diameter bearing metal-on-polymer (MoP) modular total hip replacements, which have required revision surgery due to pseudotumour formation. The failure analysis showed a discrete pattern of material loss from the distal end of the head taper/stem trunnion interface. We postulate that the use of a proximal contacting taper design had provided insufficient mechanical locking between the head and the stem, enabling the head to toggle on the trunnion. In addition, the difference in angle between the taper and the trunnion formed a crevice between the two components. Through a combination of crevice environment, mechanically assisted corrosion, mechanical wear and erosion; debris and metal-ions have been released resulting in the adverse local tissue reactions (ALTR).

Anti-biofilm performance of three natural products against initial bacterial attachment.

Marine bacteria contribute significantly towards the fouling consortium, both directly (modern foul release coatings fail to prevent "slime" attachment) and indirectly (biofilms often excrete chemical cues that attract macrofouling settlement). This study assessed the natural product anti-biofilm performance of an extract of the seaweed, Chondrus crispus, and two isolated compounds from terrestrial sources, (+)-usnic acid and juglone, against two marine biofilm forming bacteria, Cobetia marina and Marinobacter hydrocarbonoclasticus. Bioassays were developed using quantitative imaging and fluorescent labelling to test the natural products over a range of concentrations against initial bacterial attachment. All natural products affected bacterial attachment; however, juglone demonstrated the best anti-biofilm performance against both bacterial species at a concentration range between 5-20 ppm. In addition, for the first time, a dose-dependent inhibition (hormetic) response was observed for natural products against marine biofilm forming bacteria.

Electrochemical Sensing and Characterization of Aerobic Marine Bacterial Biofilms on Gold Electrode Surfaces.

Reliable and accurate in situ sensors capable of detecting and quantifying troublesome marine biofilms on metallic surfaces are increasingly necessary. A 0.2 mm diameter gold electrochemical sensor was fully characterized using cyclic voltammetry in abiotic and biotic artificial seawater media within a continuous culture flow cell to detect the growth and development of an aerobic Pseudoalteromonas sp. biofilm. Deconvolution of the abiotic and biotic responses enable the constituent extracellular electron transfer and biofilm responses to be resolved. Differentiation of enhanced oxygen reduction kinetics within the aerobic bacterial biofilm is linked to enzyme and redox mediator activities.

Experimental and computation assessment of thermomechanical effects during auxetic foam fabrication.

Auxetic foams continue to interest researchers owing to their unique and enhanced properties. Existing studies attest to the importance of fabrication mechanisms and parameters. However, disparity in thermo-mechanical parameters has left much debate as to which factors dominate fabrication output quality. This paper provides experimental, computational, and statistical insights into the mechanisms that enable auxetic foams to be produced, using key parameters reported within the literature: porosity; heating time; and volumetric compression ratio. To advance the considerations on manufacturing parameter dominance, both study design and scale have been optimised to enable statistical inferences to be drawn. Whilst being unusual for a manufacturing domain, such additional analysis provides more conclusive evidence of auxetic properties and highlights the supremacy of volumetric compression ratio in predicting Poisson's ratio outcomes in the manufacture process. Furthermore statistical results are exploited to formulate key recommendations for those wishing to maximise/optimise auxetic foam production.

Life under flow: A novel microfluidic device for the assessment of anti-biofilm technologies.

In the current study, we have developed and fabricated a novel lab-on-a-chip device for the investigation of biofilm responses, such as attachment kinetics and initial biofilm formation, to different hydrodynamic conditions. The microfluidic flow channels are designed using computational fluid dynamic simulations so as to have a pre-defined, homogeneous wall shear stress in the channels, ranging from 0.03 to 4.30 Pa, which are relevant to in-service conditions on a ship hull, as well as other man-made marine platforms. Temporal variations of biofilm formation in the microfluidic device were assessed using time-lapse microscopy, nucleic acid staining, and confocal laser scanning microscopy (CLSM). Differences in attachment kinetics were observed with increasing shear stress, i.e., with increasing shear stress there appeared to be a delay in bacterial attachment, i.e., at 55, 120, 150, and 155 min for 0.03, 0.60, 2.15, and 4.30 Pa, respectively. CLSM confirmed marked variations in colony architecture, i.e.,: (i) lower shear stresses resulted in biofilms with distinctive morphologies mainly characterised by mushroom-like structures, interstitial channels, and internal voids, and (ii) for the higher shear stresses compact clusters with large interspaces between them were formed. The key advantage of the developed microfluidic device is the combination of three architectural features in one device, i.e., an open-system design, channel replication, and multiple fully developed shear stresses.

Characterisation of Crevice and Pit Solution Chemistries Using Capillary Electrophoresis with Contactless Conductivity Detector.

The ability to predict structural degradation in-service is often limited by a lack of understanding of the evolving chemical species occurring within a range of different microenvironments associated with corrosion sites. Capillary electrophoresis (CE) is capable of analysing nanolitre solution volumes with widely disparate concentrations of ionic species, thereby producing accurate and reliable results for the analysis of the chemical compositions found within microenvironment corrosion solutions, such as those found at crevice and pit corrosion sites. In this study, CE with contactless conductivity detection (CCD) has been used to characterize pitting and crevice corrosion solution chemistries for the first time. By using the capillary electrophoresis with contactless conductivity detection (CE-CCD) system, direct and simultaneous detection of seven metal cations (Cu2+, Ni2+, Fe3+, Fe2+, Cr3+, Mn2+, and Al3+) and chloride anions was achieved with a buffer solution of 10 mM 2,6-pyridinedicarboxylic acid and 0.5 mM cetyltrimethylammonium hydroxide at pH 4 using a pre-column complexation method. The detection limits obtained for the metal cations and chloride anions were 100 and 10 ppb, respectively. The CE-CCD methodology has been demonstrated to be a versatile technique capable of speciation and quantifying the ionic species generated within artificial pit (a pencil electrode) and crevice corrosion geometries for carbon steels and nickel-aluminium bronze, thus allowing the evolution of the solution chemistry to be assessed with time and the identification of the key corrosion analyte targets for structural health monitoring.

Designing biomimetic antifouling surfaces.

Marine biofouling is the accumulation of biological material on underwater surfaces, which has plagued both commercial and naval fleets. Biomimetic approaches may well provide new insights into designing and developing alternative, non-toxic, surface-active antifouling (AF) technologies. In the marine environment, all submerged surfaces are affected by the attachment of fouling organisms, such as bacteria, diatoms, algae and invertebrates, causing increased hydrodynamic drag, resulting in increased fuel consumption, and decreased speed and operational range. There are also additional expenses of dry-docking, together with increased fuel costs and corrosion, which are all important economic factors that demand the prevention of biofouling. Past solutions to AF have generally used toxic paints or coatings that have had a detrimental effect on marine life worldwide. The prohibited use of these antifoulants has led to the search for biologically inspired AF strategies. This review will explore the natural and biomimetic AF surface strategies for marine systems.